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COMMUNICABLE DISEASES, EPIDEMIOLOGY (235 journals)                  1 2 | Last

Showing 1 - 200 of 235 Journals sorted alphabetically
ACS infectious diseases     Hybrid Journal   (Followers: 6)
Acta Parasitologica     Hybrid Journal   (Followers: 12)
Acta Tropica     Hybrid Journal   (Followers: 6)
Advances in Epidemiology     Open Access   (Followers: 8)
Advances in Infectious Diseases     Open Access   (Followers: 11)
Advances in Parasitology     Full-text available via subscription   (Followers: 6)
Advances in Pharmacoepidemiology & Drug Safety     Open Access   (Followers: 5)
Advances in Virology     Open Access   (Followers: 7)
African Journal of Infectious Diseases     Full-text available via subscription  
AIDS     Hybrid Journal   (Followers: 24)
AIDS Patient Care and STDs     Hybrid Journal   (Followers: 3)
American Journal of Epidemiology     Hybrid Journal   (Followers: 234)
American Journal of Epidemiology and Infectious Disease     Open Access   (Followers: 21)
American Journal of Infection Control     Hybrid Journal   (Followers: 36)
American Journal of Infectious Diseases     Open Access   (Followers: 14)
American Journal of Infectious Diseases and Microbiology     Open Access   (Followers: 31)
American Journal of Tropical Medicine and Hygiene     Hybrid Journal   (Followers: 8)
Annals of Epidemiology     Hybrid Journal   (Followers: 33)
Annual Review of Virology     Full-text available via subscription   (Followers: 7)
Antimicrobial Resistance and Infection Control     Open Access   (Followers: 13)
Archives of Clinical Infectious Diseases     Open Access   (Followers: 5)
Archives of Pediatric Infectious Diseases     Open Access   (Followers: 11)
Asian Journal of Epidemiology     Open Access   (Followers: 2)
Asian Journal of Research in Infectious Diseases     Open Access   (Followers: 1)
Asian Pacific Journal of Tropical Biomedicine     Open Access   (Followers: 2)
Asian Pacific Journal of Tropical Disease     Full-text available via subscription   (Followers: 3)
Asian Pacific Journal of Tropical Medicine     Open Access   (Followers: 1)
Australasian Epidemiologist     Full-text available via subscription  
Avicenna Journal of Clinical Microbiology and Infection     Open Access   (Followers: 5)
Balaba : Jurnal Litbang Pengendalian Penyakit Bersumber Binatang Banjarnegara     Open Access  
Biostatistics & Epidemiology     Hybrid Journal   (Followers: 2)
BMC Infectious Diseases     Open Access   (Followers: 21)
BMJ Global Health     Open Access   (Followers: 3)
Boletin Chileno de Parasitologia     Open Access  
Brazilian Journal of Infectious Diseases     Open Access   (Followers: 2)
Burns Open     Open Access  
Canadian Journal of Infectious Diseases and Medical Microbiology     Open Access   (Followers: 8)
Case Reports in Infectious Diseases     Open Access   (Followers: 6)
Chronic Diseases and Injuries in Canada     Free   (Followers: 1)
Chronic Diseases Journal     Open Access  
Clinical Epidemiology     Open Access   (Followers: 13)
Clinical Epidemiology and Global Health     Full-text available via subscription   (Followers: 10)
Clinical Infection in Practice     Open Access   (Followers: 2)
Clinical Infectious Diseases     Hybrid Journal   (Followers: 81)
Clinical Practice & Epidemiology in Mental Health     Open Access   (Followers: 1)
Communicable Diseases Intelligence Quarterly Report     Full-text available via subscription   (Followers: 2)
Community Acquired Infection     Open Access  
Current Epidemiology Reports     Hybrid Journal   (Followers: 8)
Current Fungal Infection Reports     Hybrid Journal   (Followers: 5)
Current HIV Research     Hybrid Journal   (Followers: 7)
Current Infectious Disease Reports     Hybrid Journal   (Followers: 5)
Current Opinion in HIV and AIDS     Hybrid Journal   (Followers: 11)
Current Opinion in Infectious Diseases     Hybrid Journal   (Followers: 25)
Current Opinion in Virology     Hybrid Journal   (Followers: 1)
Current Research in Tuberculosis     Open Access   (Followers: 3)
Current Research in Virology     Open Access   (Followers: 3)
Current Treatment Options in Infectious Diseases     Hybrid Journal   (Followers: 2)
Diagnostic Microbiology and Infectious Disease     Hybrid Journal   (Followers: 12)
Emerging Infectious Diseases     Open Access   (Followers: 32)
Emerging Infectious Diseases Journal     Open Access   (Followers: 4)
Emerging Microbes & Infections     Open Access   (Followers: 5)
Emerging Themes in Epidemiology     Open Access   (Followers: 13)
Enfermedades Infecciosas y Microbiología Clínica     Full-text available via subscription  
Environmental Epidemiology     Open Access  
Epidemiologia e Serviços de Saúde     Open Access  
Epidemiologic Methods     Hybrid Journal   (Followers: 4)
Epidemiology     Hybrid Journal   (Followers: 58)
Epidemiology & Infection     Open Access   (Followers: 22)
Epidemiology and Psychiatric Sciences     Open Access   (Followers: 4)
Epidemiology Reports     Open Access   (Followers: 3)
European Journal of Clinical Microbiology & Infectious Diseases     Hybrid Journal   (Followers: 37)
European Journal of Epidemiology     Hybrid Journal   (Followers: 35)
Experimental Parasitology     Hybrid Journal   (Followers: 2)
Expert Review of Anti-infective Therapy     Full-text available via subscription   (Followers: 4)
Food and Environmental Virology     Hybrid Journal   (Followers: 1)
Frontiers in Cellular and Infection Microbiology     Open Access   (Followers: 5)
Future Virology     Hybrid Journal   (Followers: 9)
Global Epidemiology     Open Access  
Global Journal of Rare Diseases     Open Access   (Followers: 2)
HIV Australia     Full-text available via subscription   (Followers: 3)
HIV/AIDS - Research and Palliative Care     Open Access   (Followers: 19)
Immunology and Infectious Diseases     Open Access   (Followers: 9)
Indian Journal of Tuberculosis     Full-text available via subscription  
Indonesian Journal of Tropical and Infectious Disease     Open Access  
Infectio     Open Access  
Infection     Hybrid Journal   (Followers: 9)
Infection Control and Hospital Epidemiology     Full-text available via subscription   (Followers: 48)
Infection Ecology & Epidemiology     Open Access   (Followers: 7)
Infection Prevention in Practice     Open Access   (Followers: 1)
Infection, Disease & Health     Open Access   (Followers: 9)
Infectious Disease Clinics of North America     Full-text available via subscription   (Followers: 6)
Infectious Disease Modelling     Open Access   (Followers: 2)
Infectious Disease Reports     Open Access   (Followers: 2)
Infectious Diseases     Hybrid Journal   (Followers: 7)
Infectious Diseases and Therapy     Open Access   (Followers: 2)
Infectious Diseases and Translational Medicine     Open Access   (Followers: 1)
Infectious Diseases in Clinical Practice     Hybrid Journal   (Followers: 12)
Infectious Diseases of Poverty     Open Access   (Followers: 1)
Infectious Diseases: Research and Treatment     Open Access   (Followers: 4)
Infectious Disorders - Drug Targets     Hybrid Journal  
Injury Epidemiology     Open Access   (Followers: 1)
International Journal for Parasitology     Hybrid Journal   (Followers: 11)
International Journal for Parasitology : Drugs and Drug Resistance     Open Access   (Followers: 4)
International Journal for Parasitology : Parasites and Wildlife     Open Access   (Followers: 2)
International Journal of Epidemiologic Research     Open Access   (Followers: 1)
International Journal of Epidemiology     Hybrid Journal   (Followers: 296)
International Journal of Infection     Open Access   (Followers: 2)
International Journal of Infection and Microbiology     Open Access   (Followers: 6)
International Journal of Infectious Diseases     Open Access   (Followers: 12)
International Journal of Mycobacteriology     Open Access  
International Journal of STD & AIDS     Hybrid Journal   (Followers: 7)
International Journal of Virology     Open Access   (Followers: 2)
International STD Research & Reviews     Open Access   (Followers: 1)
Iranian Journal of Epidemiology     Open Access   (Followers: 2)
JAC-Antimicrobial Resistance     Open Access   (Followers: 1)
JACC : Clinical Electrophysiology     Full-text available via subscription   (Followers: 7)
Journal Infectology     Open Access  
Journal of AIDS & Clinical Research     Open Access   (Followers: 3)
Journal of AIDS and HIV Research     Open Access   (Followers: 5)
Journal of Bamrasnaradura Infectious Diseases Institute     Open Access  
Journal of Biosafety and Biosecurity     Open Access  
Journal of Clinical Epidemiology     Hybrid Journal   (Followers: 25)
Journal of Clinical Tuberculosis and Other Mycobacterial Diseases     Open Access  
Journal of Clinical Virology     Hybrid Journal   (Followers: 18)
Journal of Epidemiological Research     Open Access   (Followers: 1)
Journal of Epidemiology & Community Health     Hybrid Journal   (Followers: 77)
Journal of Epidemiology and Global Health     Open Access   (Followers: 11)
Journal of Epidemiology and Public Health     Open Access   (Followers: 1)
Journal of General and Molecular Virology     Open Access   (Followers: 1)
Journal of Global Infectious Diseases     Open Access  
Journal of Hepatocellular Carcinoma     Open Access   (Followers: 2)
Journal of HIV and Human Reproduction     Open Access   (Followers: 4)
Journal of Hospital Infection     Hybrid Journal   (Followers: 37)
Journal of Immunological Techniques in Infectious Diseases     Hybrid Journal   (Followers: 3)
Journal of Infection     Hybrid Journal   (Followers: 7)
Journal of Infection in Developing Countries     Open Access  
Journal of Infection Prevention     Hybrid Journal   (Followers: 16)
Journal of Infectious Diseases     Hybrid Journal   (Followers: 47)
Journal of Infectious Diseases and Immunity     Open Access   (Followers: 8)
Journal of Medical Virology     Hybrid Journal   (Followers: 15)
Journal of Microbiology and Infectious Diseases     Open Access   (Followers: 2)
Journal of NeuroVirology     Hybrid Journal   (Followers: 1)
Journal of Parasitic Diseases     Hybrid Journal   (Followers: 2)
Journal of Parasitology and Vector Biology     Open Access   (Followers: 1)
Journal of Parasitology Research     Open Access   (Followers: 4)
Journal of Patient Safety & Infection Control     Open Access   (Followers: 10)
Journal of Pediatric Infectious Diseases     Hybrid Journal   (Followers: 13)
Journal of Preventive Medicine and Hygiene     Open Access   (Followers: 1)
Journal of Public Health and Epidemiology     Open Access   (Followers: 9)
Journal of Sexually Transmitted Diseases     Open Access   (Followers: 3)
Journal of the European Mosquito Control Association     Open Access  
Journal of the International Association of Providers of AIDS Care (JIAPAC)     Open Access   (Followers: 2)
Journal of the Pediatric Infectious Diseases Society     Hybrid Journal   (Followers: 11)
Journal of Tropical Medicine     Open Access   (Followers: 4)
Journal of Virology & Antiviral Research     Hybrid Journal   (Followers: 6)
Jurnal Vektor Penyakit     Open Access  
Leprosy Review     Full-text available via subscription   (Followers: 1)
Malaria Journal     Open Access   (Followers: 8)
Malaria Reports     Open Access   (Followers: 2)
Médecine et Maladies Infectieuses     Full-text available via subscription   (Followers: 1)
Medical Mycology Case Reports     Open Access  
Mediterranean Journal of Hematology and Infectious Diseases     Open Access  
Microbes and Infection     Full-text available via subscription   (Followers: 7)
Microbial Ecology in Health and Disease     Open Access   (Followers: 2)
Nepal Journal of Epidemiology     Open Access  
Nigerian Journal of Parasitology     Full-text available via subscription   (Followers: 1)
Norsk epidemiologi     Open Access   (Followers: 1)
OA Epidemiology     Open Access   (Followers: 1)
Open AIDS Journal     Open Access  
Open Epidemiology Journal     Open Access  
Open Forum Infectious Diseases     Open Access   (Followers: 4)
Open Infectious Diseases Journal     Open Access  
Open Journal of Epidemiology     Open Access   (Followers: 3)
Open Parasitology Journal     Open Access  
Open Virology Journal     Open Access  
Papillomavirus Research     Open Access  
Parasite Epidemiology and Control     Open Access  
Parasites & Vectors     Open Access   (Followers: 2)
Parasitología al día     Open Access  
Parasitologia latinoamericana     Open Access  
Parasitologists United Journal     Open Access  
Parasitology     Hybrid Journal   (Followers: 4)
Parasitology International     Hybrid Journal   (Followers: 1)
Parasitology Open     Open Access  
Parasitology Research     Hybrid Journal   (Followers: 6)
Pathogens     Open Access   (Followers: 4)
Pathogens and Global Health     Hybrid Journal   (Followers: 4)
Pediatric Infectious Disease     Full-text available via subscription   (Followers: 3)
Pediatric Infectious Disease Journal     Hybrid Journal   (Followers: 39)
Perspectives in Medical Virology     Full-text available via subscription  
Physis : Revista de Saúde Coletiva     Open Access   (Followers: 1)
PLOS Currents : Outbreaks     Open Access   (Followers: 7)
PLoS Neglected Tropical Disease     Open Access   (Followers: 19)
Preventive Medicine     Hybrid Journal   (Followers: 20)
Radiology of Infectious Diseases     Open Access   (Followers: 3)
Reports in Parasitology     Open Access   (Followers: 1)
Research and Reports in Tropical Medicine     Open Access   (Followers: 2)
Retrovirology     Open Access  
Reviews in Infection     Open Access   (Followers: 2)
Reviews in Medical Virology     Hybrid Journal   (Followers: 8)

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Journal Prestige (SJR): 1.421
Citation Impact (citeScore): 4
Number of Followers: 4  

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ISSN (Online) 2076-0817
Published by MDPI Homepage  [231 journals]
  • Pathogens, Vol. 9, Pages 597: Cyclic Hypoxia Exposure Accelerates the
           Progression of Amoebic Gill Disease

    • Authors: Tina Oldham, Tim Dempster, Philip Crosbie, Mark Adams, Barbara Nowak
      First page: 597
      Abstract: Amoebic gill disease (AGD), caused by the amoeba Neoparamoeba perurans, has led to considerable economic losses in every major Atlantic salmon producing country, and is increasing in frequency. The most serious infections occur during summer and autumn, when temperatures are high and poor dissolved oxygen (DO) conditions are most common. Here, we tested if exposure to cyclic hypoxia at DO saturations of 40–60% altered the course of infection with N. perurans compared to normoxic controls maintained at ≥90% DO saturation. Although hypoxia exposure did not increase initial susceptibility to N. perurans, it accelerated progression of the disease. By 7 days post-inoculation, amoeba counts estimated from qPCR analysis were 1.7 times higher in the hypoxic treatment than in normoxic controls, and cumulative mortalities were twice as high (16 ± 4% and 8 ± 2%), respectively. At 10 days post-inoculation, however, there were no differences between amoeba counts in the hypoxic and normoxic treatments, nor in the percentage of filaments with AGD lesions (control = 74 ± 2.8%, hypoxic = 69 ± 3.3%), or number of lamellae per lesion (control = 30 ± 0.9%, hypoxic = 27.9 ± 0.9%) as determined by histological examination. Cumulative mortalities at the termination of the experiment were similarly high in both treatments (hypoxic = 60 ± 2%, normoxic = 53 ± 11%). These results reveal that exposure to cyclic hypoxia in a diel pattern, equivalent to what salmon are exposed to in marine aquaculture cages, accelerated the progression of AGD in post-smolts.
      Citation: Pathogens
      PubDate: 2020-07-22
      DOI: 10.3390/pathogens9080597
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 598: Correlation of Feline Coronavirus Shedding
           in Feces with Coronavirus Antibody Titer

    • Authors: Sandra Felten, Ute Klein-Richers, Regina Hofmann-Lehmann, Michèle Bergmann, Stefan Unterer, Christian M. Leutenegger, Katrin Hartmann
      First page: 598
      Abstract: Background: Feline coronavirus (FCoV) infection is ubiquitous in multi-cat households. Responsible for the continuous presence are cats that are chronically shedding a high load of FCoV. The aim of the study was to determine a possible correlation between FCoV antibody titer and frequency and load of fecal FCoV shedding in cats from catteries. Methods: Four fecal samples from each of 82 cats originating from 19 German catteries were examined for FCoV viral loads by quantitative reverse transcriptase polymerase chain reaction (RT-qPCR). Additionally, antibody titers were determined by an immunofluorescence assay. Results: Cats with antibodies were more likely to be FCoV shedders than non-shedders, and there was a weak positive correlation between antibody titer and mean fecal virus load (Spearman r = 0.2984; p = 0.0072). Antibody titers were significantly higher if cats shed FCoV more frequently throughout the study period (p = 0.0063). When analyzing only FCoV shedders, cats that were RT-qPCR-positive in all four samples had significantly higher antibody titers (p = 0.0014) and significantly higher mean fecal virus loads (p = 0.0475) than cats that were RT-qPCR-positive in only one, two, or three samples. Conclusions: The cats’ antibody titers correlate with the likelihood and frequency of FCoV shedding and fecal virus load. Chronic shedders have higher antibody titers and shed more virus. This knowledge is important for the management of FCoV infections in multi-cat environments, but the results indicate that antibody measurement cannot replace fecal RT-qPCR.
      Citation: Pathogens
      PubDate: 2020-07-22
      DOI: 10.3390/pathogens9080598
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 599: Heterologous Expression of the
           Pathogen-Specific LIC11711 Gene in the Saprophyte L. biflexa Increases
           Bacterial Binding to Laminin and Plasminogen

    • Authors: Leandro Toshio Kochi, Luis Guilherme Virgílio Fernandes, Ana Lucia Tabet Oller Nascimento
      First page: 599
      Abstract: Leptospirosis is a febrile disease and the etiological agents are pathogenic bacteria of the genus Leptospira. The leptospiral virulence mechanisms are not fully understood and the application of genetic tools is still limited, despite advances in molecular biology techniques. The leptospiral recombinant protein LIC11711 has shown interaction with several host components, indicating a potential function in virulence. This study describes a system for heterologous expression of the L. interrogans gene lic11711 using the saprophyte L. biflexa serovar Patoc as a surrogate, aiming to investigate its possible activity in bacterial virulence. Heterologous expression of LIC11711 was performed using the pMaOri vector under regulation of the lipL32 promoter. The protein was found mainly on the leptospiral outer surface, confirming its location. The lipL32 promoter enhanced the expression of LIC11711 in L. biflexa compared to the pathogenic strain, indicating that this strategy may be used to overexpress low-copy proteins. The presence of LIC11711 enhanced the capacity of L. biflexa to adhere to laminin (Lam) and plasminogen (Plg)/plasmin (Pla) in vitro, suggesting the involvement of this protein in bacterial pathogenesis. We show for the first time that the expression of LIC11711 protein of L. interrogans confers a virulence-associated phenotype on L. biflexa, pointing out possible mechanisms used by pathogenic leptospires.
      Citation: Pathogens
      PubDate: 2020-07-22
      DOI: 10.3390/pathogens9080599
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 600: Synthetic Peptides as a Promising
           Alternative to Control Viral Infections in Atlantic Salmon

    • Authors: Constanza Cárdenas, Fanny Guzmán, Marisela Carmona, Cristian Muñoz, Luis Nilo, Alvaro Labra, Sergio H. Marshall
      First page: 600
      Abstract: Viral infections in salmonids represent an ongoing challenge for the aquaculture industry. Two RNA viruses, the infectious pancreatic necrosis virus (IPNV) and the infectious salmon anemia virus (ISAV), have become a latent risk without healing therapies available for either. In this context, antiviral peptides emerge as effective and relatively safe therapeutic molecules. Based on in silico analysis of VP2 protein from IPNV and the RNA-dependent RNA polymerase from ISAV, a set of peptides was designed and were chemically synthesized to block selected key events in their corresponding infectivity processes. The peptides were tested in fish cell lines in vitro, and four were selected for decreasing the viral load: peptide GIM182 for IPNV, and peptides GIM535, GIM538 and GIM539 for ISAV. In vivo tests with the IPNV GIM 182 peptide were carried out using Salmo salar fish, showing a significant decrease of viral load, and proving the safety of the peptide for fish. The results indicate that the use of peptides as antiviral agents in disease control might be a viable alternative to explore in aquaculture.
      Citation: Pathogens
      PubDate: 2020-07-23
      DOI: 10.3390/pathogens9080600
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 601: Strongyloidiasis in Southern Alicante
           (Spain): Comparative Retrospective Study of Autochthonous and Imported

    • Authors: Ana Lucas Dato, María Isabel Pacheco-Tenza, Emilio Borrajo Brunete, Belén Martínez López, María García López, Inmaculada González Cuello, Joan Gregori Colomé, María Navarro Cots, José María Saugar, Elisa García-Vazquez, José Antonio Ruiz-Maciá, Jara Llenas-García
      First page: 601
      Abstract: Background: Strongyloidiasis is a parasitic disease with global prevalence. In Spain, autochthonous cases are concentrated in the Mediterranean basin. We aimed to analyze clinical and epidemiological characteristics of Strongyloides stercoralis infection in Vega Baja del Segura (Spain), comparing autochthonous versus imported cases. Methods: Observational retrospective study of all strongyloidiasis cases from January 2009 to January 2019. Cases were diagnosed by stool larvae visualization, positive culture, PCR, Strongyloides serology, and/or compatible histology. Results: We included 36 patients (21 men) with a mean age of 60.8 years ±17.6; 15 cases were autochthonous and 21 imported 80.9% from Latin America. Autochthonous cases were associated with older age (mean 71.3 vs. 53.3 years; p = 0.002), male sex (odds ratio (OR) 5.33; 95% confidence interval (CI) 1.15–24.68; p = 0.041), and agricultural activity (OR 13.5; 95% CI 2.4–73.7; p = 0.002). Fourteen were asymptomatic, three autochthonous cases presented with hyperinfection syndrome, and two patients died. There was no difference between autochthonous versus imported origin in eosinophilia at diagnosis (93.3% vs. 75%; p = 0.207), treatment received, or clinical response (85.7% vs. 88.9% cured; p = 1). Conclusion: In our region, imported strongyloidiasis coexists with autochthonous cases, which are mainly in older male farmers who are diagnosed at more advanced stages. Systematic screening programs are needed.
      Citation: Pathogens
      PubDate: 2020-07-23
      DOI: 10.3390/pathogens9080601
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 602: Pathogenesis of Fungal and Bacterial

    • Authors: Jennifer Geddes-McAlister
      First page: 602
      Abstract: The pathogenesis of fungal and bacterial microbes is a complex process involving distinct parameters, including virulence factors, nutrient sensing and availability, microbial signals, as well as host status and defense responses. Defining pathogenesis improves our understanding of how an organism causes diseases and provides insight into novel prospects to combat infection. The effects of pathogenic microbes have significant impact on diverse sectors, including health, agriculture, and economics, underscoring their immense importance in society. Articles in this Special Issue address unique aspects of microbial pathogenesis by exploring interactions between host and pathogen during infection, defining inflammatory immune responses, profiling the importance of essential microbial structures associated with virulence, and outlining critical considerations driving complex diseases.
      Citation: Pathogens
      PubDate: 2020-07-23
      DOI: 10.3390/pathogens9080602
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 603: FCoV Viral Sequences of Systemically
           Infected Healthy Cats Lack Gene Mutations Previously Linked to the
           Development of FIP

    • Authors: Mirjam Lutz, Aline R. Steiner, Valentino Cattori, Regina Hofmann-Lehmann, Hans Lutz, Anja Kipar, Marina L. Meli
      First page: 603
      Abstract: Feline infectious peritonitis (FIP)—the deadliest infectious disease of young cats in shelters or catteries—is induced by highly virulent feline coronaviruses (FCoVs) emerging in infected hosts after mutations of less virulent FCoVs. Previous studies have shown that some mutations in the open reading frames (ORF) 3c and 7b and the spike (S) gene have implications for the development of FIP, but mainly indirectly, likely also due to their association with systemic spread. The aim of the present study was to determine whether FCoV detected in organs of experimentally FCoV infected healthy cats carry some of these mutations. Viral RNA isolated from different tissues of seven asymptomatic cats infected with the field strains FCoV Zu1 or FCoV Zu3 was sequenced. Deletions in the 3c gene and mutations in the 7b and S genes that have been shown to have implications for the development of FIP were not detected, suggesting that these are not essential for systemic viral dissemination. However, deletions and single nucleotide polymorphisms leading to truncations were detected in all nonstructural proteins. These were found across all analyzed ORFs, but with significantly higher frequency in ORF 7b than ORF 3a. Additionally, a previously unknown homologous recombination site was detected in FCoV Zu1.
      Citation: Pathogens
      PubDate: 2020-07-24
      DOI: 10.3390/pathogens9080603
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 604: Health Surveillance of Wild Brown Trout
           (Salmo trutta fario) in the Czech Republic Revealed a Coexistence of
           Proliferative Kidney Disease and Piscine Orthoreovirus-3 Infection

    • Authors: Pojezdal, Adamek, Syrová, Steinhagen, Minářová, Papežíková, Seidlová, Reschová, Palíková
      First page: 604
      Abstract: The population of brown trout (Salmo trutta fario) in continental Europe is on the decline, with infectious diseases confirmed as one of the causative factors. However, no data on the epizootiological situation of wild fish in the Czech Republic are currently available. In this study, brown trout (n = 260) from eight rivers were examined for the presence of viral and parasitical pathogens. Salmonid alphavirus-2, infectious pancreatic necrosis virus, piscine novirhabdovirus (VHSV) and salmonid novirhabdovirus (IHNV) were not detected using PCR. Cell culturing showed no viruses as well, and serological analysis of 110 sera did not detect any specific antibodies against VHSV or IHNV. Fish from two rivers were positive for the presence of piscine orthoreovirus-3 (PRV-3), subtype PRV-3b. However, none of the PRV-3-positive fish showed gross pathologies typically associated with PRV infections. By far the most widespread pathogen was Tetracapsuloides bryosalmonae which was confirmed in each of the examined locations, with a prevalence of up to 65% and 100%, as established by immunohistochemistry and PCR, respectively. Furthermore, up to 43.8% of fish showed signs of proliferative kidney disease caused by T. bryosalmonae, suggesting that this parasite is a main health challenge for brown trout in the Czech Republic.
      Citation: Pathogens
      PubDate: 2020-07-24
      DOI: 10.3390/pathogens9080604
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 605: Control of Persistent Salmonella Infection
           Relies on Constant Thymic Output Despite Increased Peripheral
           Antigen-Specific T Cell Immunity

    • Authors: J. Alan Goggins, Jonathan R Kurtz, James B. McLachlan
      First page: 605
      Abstract: Recent thymic emigrants are the youngest subset of peripheral T cells and their involvement in combating persistent bacterial infections has not been explored. Here, we hypothesized that CD4+ recent thymic emigrants are essential immune mediators during persistent Salmonella infection. To test this, we thymectomized adult mice either prior to, or during, persistent Salmonella infection. We found that thymic output is crucial in the formation of protective immune responses during the early formation of a Salmonella infection but is dispensable once persistent Salmonella infection is established. Further, we show that thymectomized mice demonstrate increased infection-associated mortality and bacterial burdens. Unexpectedly, numbers of Salmonella-specific CD4+ T cells were significantly increased in thymectomized mice compared to sham control mice. Lastly, we found that T cells from thymectomized mice may be impaired in producing the effector cytokine IL-17 at early time points of infection, compared to thymically intact mice. Together, these results imply a unique role for thymic output in the formation of immune responses against a persistent, enteric pathogen.
      Citation: Pathogens
      PubDate: 2020-07-25
      DOI: 10.3390/pathogens9080605
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 606: Molecular Characterization and Pathogenicity
           of Chicken Parvovirus (ChPV) in Specific Pathogen-Free Chicks Infected

    • Authors: Luis Fabian N. Nuñez, Silvana H. Santander-Parra, David I. De la Torre, Lilian R. M. de Sá, Marcos R. Buim, Claudete S. Astolfi-Ferreira, Antonio J. Piantino Ferreira
      First page: 606
      Abstract: Chicken parvovirus (ChPV) is an agent frequently associated with runting stunting syndrome (RSS). This syndrome has been reported in association with ChPV in many countries, including Brazil; however, studies characterizing the virus on a molecular level are scarce, and ChPV pathogenicity in day-old chicks remains unclear. The aim of the present work was to establish the molecular characteristics of ChPV, determine the pathogenicity of ChPV in SPF chicks and detect and quantify ChPV by qPCR in several tissues and chicks of different ages. The experimental challenge was performed at one day of age, and daily and weekly observations were performed and five birds from each experimental group (mock and infected birds) were euthanized to perform the different analysis. ChPV genome copies were detected and quantified by qPCR in gut, spleen, thymus, kidney, pancreas, proventriculus and bursa. Clinically, the infected group presented with diarrhea 24 h post-infection, which persisted until 42 days of age. The small intestine was distended, and its contents were aqueous and foamy. Enteritis and dilated crypts with cyst shapes were observed in intestinal segments. Acute pancreatitis associated with lymphocytic nodules, infiltrating lymphocytes and plasma cells between the pancreatic acinus was observed. Koch’s postulate was demonstrated and the genetic characterization of the VP1 gene showed that the Brazilian ChPV isolate belongs to the ChPV II group.
      Citation: Pathogens
      PubDate: 2020-07-25
      DOI: 10.3390/pathogens9080606
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 607: MCMV Dissemination from Latently-Infected
           Allografts Following Transplantation into Pre-Tolerized Recipients

    • Authors: Sahil Shah, Matthew DeBerge, Andre Iovane, Shixian Yan, Longhui Qiu, Jiao-Jing Wang, Yashpal S. Kanwar, Mary Hummel, Zheng J. Zhang, Michael M. Abecassis, Xunrong Luo, Edward B. Thorp
      First page: 607
      Abstract: Transplantation tolerance is achieved when recipients are unresponsive to donor alloantigen yet mobilize against third-party antigens, including virus. After transplantation, cytomegalovirus (CMV) reactivation in latently-infected transplants reduces allograft viability. To determine if pre-tolerized recipients are resistant to viral dissemination in this setting, we transfused chemically-fixed donor splenocytes (1-ethyl-3- (3′-dimethyl-aminopropyl)-carbo-diimide (ECDI)-treated splenocytes (ECDIsp)) to induce donor antigen tolerance without immunosuppression. In parallel, we implanted donor islet cells to validate operational tolerance. These pre-tolerized recipients were implanted with murine CMV (MCMV) latently-infected donor kidneys (a validated model of CMV latency) to monitor graft inflammation and viral dissemination. Our results indicate that tolerance to donor islets was sustained in recipients after implantation of donor kidneys. In addition, kidney allografts implanted after ECDIsp and islet implantation exhibited low levels of fibrosis and tubulitis. In contrast, kidney cellular and innate immune infiltrates trended higher in the CMV group and exhibited increased markers of CD8+ T cell activation. Tolerance induction was unable to prevent increases in MCMV-specific CD8+ T cells or dissemination of viral IE-1 DNA. Our data suggest that latently-infected allografts are inherently more susceptible to inflammation that is associated with viral dissemination in pre-tolerized recipients. Thus, CMV latently-infected allografts require enhanced strategies to protect allograft integrity and viral spread.
      Citation: Pathogens
      PubDate: 2020-07-26
      DOI: 10.3390/pathogens9080607
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 608: Protection of Piglets with Maternally
           Derived Antibodies from Sows Inoculated with an Attenuated Live Marker
           Classical Swine Fever Vaccine (Flc-LOM-BErns)

    • Authors: SeEun Choe, Jihye Shin, Ki-Sun Kim, Sok Song, Ra Mi Cha, Byung-Il Jung, Bang-Hun Hyun, Bong-Kyun Park, Dong-Jun An
      First page: 608
      Abstract: Here, we investigated the protective efficacy provided by passive immunity induced by a classical swine fever (Flc-LOM-BErns) vaccine with the newly developed DIVA (Differentiating Infected from Vaccinated Animals) function. Ten pigs (aged 40–60 days) with maternally derived antibodies (MDAs) obtained from sows inoculated with the Flc-LOM-BErns vaccine were challenged with virulent classical swine fever virus (CSFV). Pigs with an MDA titer of 6 log2 induced by the Flc-LOM-BErns vaccine were fully protected against virulent CSFV challenge but not the pigs with an MDA titer under 5 log2. In addition, Flc-LOM-BErns vaccine-derived MDAs successfully differentiated vaccinated pigs by bovine viral diarrhea virus (BVDV) Erns/CSFV Erns antibody detection, functioning as a DIVA.
      Citation: Pathogens
      PubDate: 2020-07-27
      DOI: 10.3390/pathogens9080608
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 609: Zebrafish as a Model for Fish Diseases in

    • Authors: Louise von Gersdorff Jørgensen
      First page: 609
      Abstract: The use of zebrafish as a model for human conditions is widely recognized. Within the last couple of decades, the zebrafish has furthermore increasingly been utilized as a model for diseases in aquacultured fish species. The unique tools available in zebrafish present advantages compared to other animal models and unprecedented in vivo imaging and the use of transgenic zebrafish lines have contributed with novel knowledge to this field. In this review, investigations conducted in zebrafish on economically important diseases in aquacultured fish species are included. Studies are summarized on bacterial, viral and parasitic diseases and described in relation to prophylactic approaches, immunology and infection biology. Considerable attention has been assigned to innate and adaptive immunological responses. Finally, advantages and drawbacks of using the zebrafish as a model for aquacultured fish species are discussed.
      Citation: Pathogens
      PubDate: 2020-07-27
      DOI: 10.3390/pathogens9080609
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 610: Mycobacterium pseudoshottsii in
           Mediterranean Fish Farms: New Trouble for European Aquaculture'

    • Authors: Davide Mugetti, Katia Varello, Andrea Gustinelli, Paolo Pastorino, Vasco Menconi, Daniela Florio, Maria Letizia Fioravanti, Elena Bozzetta, Simona Zoppi, Alessandro Dondo, Marino Prearo
      First page: 610
      Abstract: Mycobacterium pseudoshottsii, a slow-growing mycobacterium closely related to M. marinum, has been isolated only in wild fish in the United States and in Japanese fish farms to date. Here, we report cases of mortality in three farmed fish species (Dicentrarchus labrax, Sparus aurata, and Sciaenops ocellatus) caused by M. pseudoshottsii in Italy. Samples underwent necropsy, histology, and culture with pathogen identification based on PCR and sequencing of housekeeping genes (16S rRNA, hsp65, rpoB). Multifocal to coalescing granulomatous and necrotizing inflammation with acid-fast bacilli were observed in the parenchymatous organs, from which M. pseudoshottsii was isolated and identified. Phylogenetic analysis confirmed the results of gene sequencing and allowed subdivision of the isolates into three distinct groups. M. pseudoshottsii poses a potential threat for Mediterranean aquaculture. Its origin in the area under study needs to be clarified, as well as the threat to the farmed fish species.
      Citation: Pathogens
      PubDate: 2020-07-27
      DOI: 10.3390/pathogens9080610
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 611: Specific Detection of Yersinia pestis Based
           on Receptor Binding Proteins of Phages

    • Authors: Friederike Born, Peter Braun, Holger C. Scholz, Gregor Grass
      First page: 611
      Abstract: The highly pathogenic bacterium Yersinia pestis is the causative agent of plague, a notorious infectious zoonotic disease. When transmitted from person to person as pneumonic plague via droplets, Y. pestis is highly contagious and in most cases is fatal if left untreated. Thus, when plague is suspected, rapid diagnosis is crucial, as a serious course of the infection is only averted by early antibiotic therapy. The bacterium is easy to cultivate, accessible and has a high potential for nefarious use such as bioterrorism. Highly specific, rapid and easy-to-use confirmatory diagnostic methods are required to reliably identify the pathogen independently from PCR-based methods or F1 antigen-based immunological detection. Yersinia pestis specific phages such as L-413C and ΦA1122 are already used for detection of Y. pestis in bacterial plaque or biosensor assays. Here, we made use of the host specificities conferred by phage receptor binding (or tail fiber/spike) proteins (RBP) for developing a specific, fast and simple fluorescence-microscopy-based detection method for Y. pestis. Genes of putative RBP of phages L-413C (gpH) and ΦA1122 (gp17) were fused with those of fluorescent proteins and recombinant receptor-reporter fusion proteins were produced heterologously in Escherichia coli. When first tested on attenuated Y. pestis strain EV76, RBP-reporters bound to the bacterial cell surface. This assay could be completed within a few minutes using live or formaldehyde-inactivated cells. Specificity tests using cultures of closely related Yersinia species and several inactivated fully virulent Y. pestis strains exhibited high specificities of the RBP-reporters against Y. pestis. The L-413C RBP proved to be especially specific, as it only detected Y. pestis at all temperatures tested, whereas the RBP of ΦA1122 also bound to Y. pseudotuberculosis strains at 37 °C (but not at 28, 20 or 6 °C). Finally, the Y. pestis-specific capsule, produced when grown at 37 °C, significantly reduced binding of phage ΦA1122 RBP, whereas the capsule only slightly diminished binding of L-413C RBP.
      Citation: Pathogens
      PubDate: 2020-07-27
      DOI: 10.3390/pathogens9080611
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 612: Dolphins Stranded along the Tuscan Coastline
           (Central Italy) of the “Pelagos Sanctuary”: A Parasitological

    • Authors: Giuliana Terracciano, Gianluca Fichi, Antonia Comentale, Enrica Ricci, Cecilia Mancusi, Stefania Perrucci
      First page: 612
      Abstract: Parasite monitoring is considered a necessary step for cetacean management and conservation. Between February 2013 and July 2015, 26 dolphins (15 Stenella coeruleoalba, 10 Tursiops truncatus, and one Grampus griseus) stranded along the Tuscan coastline of the protected marine area “Pelagos Sanctuary”, were examined. Organs, tissues, and faecal and blood samples taken from all animals were analysed by parasitological, immunological, and molecular techniques. Twenty-one out of 26 dolphins (80.77%) tested positive for at least one parasite species, and 13/15 (86.7%) S. coeruleoalba, 7/10 (70%) T. truncatus, and the single G. griseus were found positive. Identified parasites included the nematodes Skrjabinalius guevarai (7.69%, 2/26), Halocercus lagenorhynchi (3.85%, 1/26), Halocercus delphini (7.69%, 2/26), Stenurus ovatus (7.69%, 2/26), Crassicauda spp. (7.69%, 2/26); the trematodes Pholeter gastrophilus (26.92%, 7/26), Campula palliata (3.85%, 1/26); the cestodes Phyllobothrium delphini (42.31%, 11/26), Monorygma grimaldii (23.08%, 6/26), Tetrabothrium forsteri (7.69%, 2/26), Strobilocephalus triangularis (7.69%, 2/26), and the acanthocephalan Bolbosoma vasculosum (7.69%, 2/26). Moreover, 6/26 (23%) animals scored positive to Toxoplasma gondii at serology, but PCR confirmed the infection (T. gondii Type II genotype) in a single animal. In examined dolphins, obtained results showed a high prevalence of endoparasites, which included species considered as a cause of severe debilitation or death.
      Citation: Pathogens
      PubDate: 2020-07-27
      DOI: 10.3390/pathogens9080612
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 613: Effectiveness of Dunaliella salina Extracts
           against Bacillus subtilis and Bacterial Plant Pathogens

    • Authors: Alfredo Ambrico, Mario Trupo, Rosaria Magarelli, Roberto Balducchi, Angelo Ferraro, Evangelos Hristoforou, Tiziana Marino, Dino Musmarra, Patrizia Casella, Antonio Molino
      First page: 613
      Abstract: Several bacteria pathogens are responsible for plant diseases causing significant economic losses. The antibacterial activity of Dunaliella salina microalgae extracts were investigated in vitro and in vivo. First, biomass composition was chemically characterized and subjected to extraction using polar/non-polar solvents. The highest extraction yield was obtained using chloroform:methanol (1:1 v/v) equal to 170 mg g−1 followed by ethanol (88 mg g−1) and hexane (61 mg g−1). In vitro examination of hexane extracts of Dunaliella salina demonstrated antibacterial activity against all tested bacteria. The hexane extract showed the highest amount of β-carotene with respect to the others, so it was selected for subsequent analyses. In vivo studies were also carried out using hexane extracts of D. salina against Pseudomonas syringae pv. tomato and Pectobacterium carotovorum subsp. carotovorum on young tomato plants and fruits of tomato and zucchini, respectively. The treated young tomato plants exhibited a reduction of 65.7% incidence and 77.0% severity of bacterial speck spot disease. Similarly, a reduction of soft rot symptoms was observed in treated tomato and zucchini fruits with a disease incidence of 5.3% and 12.6% with respect to 90.6% and 100%, respectively, for the positive control.
      Citation: Pathogens
      PubDate: 2020-07-28
      DOI: 10.3390/pathogens9080613
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 614: Modulation of Host Lipid Pathways by
           Pathogenic Intracellular Bacteria

    • Authors: Paige E. Allen, Juan J. Martinez
      First page: 614
      Abstract: Lipids are a broad group of molecules required for cell maintenance and homeostasis. Various intracellular pathogens have developed mechanisms of modulating and sequestering host lipid processes for a large array of functions for both bacterial and host cell survival. Among the host cell lipid functions that intracellular bacteria exploit for infection are the modulation of host plasma membrane microdomains (lipid rafts) required for efficient bacterial entry; the recruitment of specific lipids for membrane integrity of intracellular vacuoles; and the utilization of host lipid droplets for the regulation of immune responses and for energy production through fatty acid β-oxidation and oxidative phosphorylation. The majority of published studies on the utilization of these host lipid pathways during infection have focused on intracellular bacterial pathogens that reside within a vacuole during infection and, thus, have vastly different requirements for host lipid metabolites when compared to those intracellular pathogens that are released into the host cytosol upon infection. Here we summarize the mechanisms by which intracellular bacteria sequester host lipid species and compare the modulation of host lipid pathways and metabolites during host cell infection by intracellular pathogens residing in either a vacuole or within the cytosol of infected mammalian cells. This review will also highlight common and unique host pathways necessary for intracellular bacterial growth that could potentially be targeted for therapeutic intervention.
      Citation: Pathogens
      PubDate: 2020-07-28
      DOI: 10.3390/pathogens9080614
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 615: Flash-Like Albuminuria in Acute Kidney
           Injury Caused by Puumala Hantavirus Infection

    • Authors: Paula Mantula, Johanna Tietäväinen, Jan Clement, Onni Niemelä, Ilkka Pörsti, Antti Vaheri, Jukka Mustonen, Satu Mäkelä, Tuula Outinen
      First page: 615
      Abstract: Transient proteinuria and acute kidney injury (AKI) are characteristics of Puumala virus (PUUV) infection. Albuminuria peaks around the fifth day and associates with AKI severity. To evaluate albuminuria disappearance rate, we quantified albumin excretion at different time points after the fever onset. The study included 141 consecutive patients hospitalized due to acute PUUV infection in Tampere University Hospital, Finland. Timed overnight albumin excretion (cU-Alb) was measured during the acute phase in 133 patients, once or twice during the convalescent phase within three months in 94 patients, and at six months in 36 patients. During hospitalization, 30% of the patients had moderately increased albuminuria (cU-Alb 20–200 μg/min), while 57% presented with severely increased albuminuria (cU-Alb >200 μg/min). Median cU-Alb was 311 μg/min (range 2.2–6460) ≤7 days after fever onset, 235 μg/min (range 6.8–5479) at 8–13 days and 2.8 μg/min (range 0.5–18.2) at 14–20 days. After that, only one of the measurements showed albuminuria (35.4 μg/min at day 44). At six months, the median cU-Alb was 2.0 μg/min (range 0.6–14.5). Albuminuria makes a flash-like appearance in PUUV infection and returns rapidly to normal levels within 2–3 weeks after fever onset. In the case of AKI, this is a unique phenomenon.
      Citation: Pathogens
      PubDate: 2020-07-28
      DOI: 10.3390/pathogens9080615
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 616: Ticks and Tick-Borne Diseases in Cuba, Half
           a Century of Scientific Research

    • Authors: Dasiel Obregón Alvarez, Belkis Corona-González, Alina Rodríguez-Mallón, Islay Rodríguez Gonzalez, Pastor Alfonso, Angel A. Noda Ramos, Adrian A. Díaz-Sánchez, Maylin González Navarrete, Rafmary Rodríguez Fernández, Luis Méndez Mellor, Helen N. Catanese, Manuel Peláez, Yousmel Alemán Gainza, Roxana Marrero-Perera, Lisset Roblejo-Arias, Evelyn Lobo-Rivero, Claudia B. Silva, Adivaldo H. Fonseca, Eugenio Roque López, Alejandro Cabezas-Cruz
      First page: 616
      Abstract: Ticks and the vast array of pathogens they transmit, including bacteria, viruses, protozoa, and helminths, constitute a growing burden for human and animal health worldwide. In Cuba, the major tropical island in the Caribbean, ticks are an important cause of vector-borne diseases affecting livestock production, pet animal health and, to a lesser extent, human health. The higher number of tick species in the country belong to the Argasidae family and, probably less known, is the presence of an autochthonous tick species in the island, Ixodes capromydis. Herein, we provide a comprehensive review of the ticks and tick-borne pathogens (TBPs) affecting animal and human health in Cuba. The review covers research results including ecophysiology of ticks, the epidemiology of TBPs, and the diagnostic tools used currently in the country for the surveillance of TBPs. We also introduce the programs implemented in the country for tick control and the biotechnology research applied to the development of anti-tick vaccines.
      Citation: Pathogens
      PubDate: 2020-07-28
      DOI: 10.3390/pathogens9080616
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 617: Ivermectin Treatment Response in Onchocerca
           Volvulus Infected Persons with Epilepsy: A Three-Country Short Cohort

    • Authors: Alfred Dusabimana, Dan Bhwana, Stephen Raimon, Bruno P. Mmbando, An Hotterbeekx, Floribert Tepage, Michel Mandro, Joseph N. Siewe Fodjo, Steven Abrams, Robert Colebunders
      First page: 617
      Abstract: Despite a long history of community-directed treatment with ivermectin (CDTI), a high ongoing Onchocerca volvulus transmission is observed in certain onchocerciasis-endemic regions in Africa with a high prevalence of epilepsy. We investigated factors associated with higher microfilarial (mf) density after ivermectin treatment. Skin snips were obtained from O. volvulus-infected persons with epilepsy before, and 3 to 5 months after ivermectin treatment. Participants were enrolled from 4 study sites: Maridi (South Sudan); Logo and Aketi (Democratic Republic of Congo); and Mahenge (Tanzania). Of the 329 participants, 105 (31.9%) had a post-treatment mf density >20% of the pre-treatment value. The percentage reduction in the geometric mean mf density ranged from 69.0% (5 months after treatment) to 89.4% (3 months after treatment). A higher pre-treatment mf density was associated with increased probability of a positive skin snip after ivermectin treatment (p = 0.016). For participants with persistent microfiladermia during follow-up, a higher number of previous CDTI rounds increased the odds of having a post-treatment mf density >20% of the pre-treatment value (p = 0.006). In conclusion, the high onchocerciasis transmission in the study sites may be due to initially high infection intensity in some individuals. Whether the decreasing effect of ivermectin with increasing years of CDTI results from sub-optimal response mechanisms warrants further research.
      Citation: Pathogens
      PubDate: 2020-07-29
      DOI: 10.3390/pathogens9080617
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 618: Immune Control of Herpesvirus Infection in

    • Authors: Jacinta R Agius, Serge Corbeil, Karla J Helbig
      First page: 618
      Abstract: Molluscan herpesviruses that are capable of infecting economically important species of abalone and oysters have caused significant losses in production due to the high mortality rate of infected animals. Current methods in preventing and controlling herpesviruses in the aquacultural industry are based around biosecurity measures which are impractical and do not contain the virus as farms source their water from oceans. Due to the lack of an adaptive immune system in molluscs, vaccine related therapies are not a viable option; therefore, a novel preventative strategy known as immune priming was recently explored. Immune priming has been shown to provide direct protection in oysters from Ostreid herpesvirus-1, as well as to their progeny through trans-generational immune priming. The mechanisms of these processes are not completely understood, however advancements in the characterisation of the oyster immune response has assisted in formulating potential hypotheses. Limited literature has explored the immune response of abalone infected with Haliotid herpesvirus as well as the potential for immune priming in these species, therefore, more research is required in this area to determine whether this is a practical solution for control of molluscan herpesviruses in an aquaculture setting.
      Citation: Pathogens
      PubDate: 2020-07-29
      DOI: 10.3390/pathogens9080618
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 619: Newcastle Disease Virus as a Vaccine Vector
           for SARS-CoV-2

    • Authors: Edris Shirvani, Siba K. Samal
      First page: 619
      Abstract: The emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has resulted in more than 16 million infections and more than 600,000 deaths worldwide. There is an urgent need to develop a safe and effective vaccine against SARS-CoV-2. Currently, several strategies are being pursued to develop a safe and effective SARS-CoV-2 vaccine. However, each vaccine strategy has distinct advantages and disadvantages. Therefore, it is important to evaluate multiple vaccine platforms to select the most efficient vaccine platform for SARS-CoV-2. In this regard, Newcastle disease virus (NDV), an avian virus, has several well-suited properties for development of a vector vaccine against SARS-CoV-2. Here, we elaborate on the idea of considering NDV as a vaccine vector for SARS-CoV-2.
      Citation: Pathogens
      PubDate: 2020-07-29
      DOI: 10.3390/pathogens9080619
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 620: Bovine Polyomavirus 2 is a Probable Cause of
           Non-Suppurative Encephalitis in Cattle

    • Authors: Melanie M. Hierweger, Michel C. Koch, Torsten Seuberlich
      First page: 620
      Abstract: Tissues from two cows with neurological signs that were admitted to the Vetsuisse Faculty under suspicion of rabies and bovine spongiform encephalopathy (BSE), respectively, were further analyzed for this case report. After histopathological examination and exclusion of BSE and rabies, the animals were diagnosed with etiologically unresolved disseminated non-suppurative encephalitis. Using next-generation sequencing, we detected the full genome of bovine polyomavirus 2 (BoPyV2) in brain samples from both animals. This virus has been identified in beef samples in three independent studies conducted in the United States and Germany, but has not been linked to any disease. Analysis of the two new BoPyV2 genome sequences revealed close phylogenetic relationships to one another and to BoPyV2 isolates detected in beef samples. In situ hybridization demonstrated the presence of viral nucleic acid in all investigated brain areas and in areas with signs of inflammation in both animals. Thus, we provide the first evidence that BoPyV2 is a probable cause of non-suppurative encephalitis in cattle, and encourage further molecular and serological testing to elucidate the disease’s epidemiology, as well as experimental transmission studies to prove causality between the infection and disease.
      Citation: Pathogens
      PubDate: 2020-07-29
      DOI: 10.3390/pathogens9080620
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 621: Prohibitin 2 is Involved in Parkin-Mediated
           Mitophagy in Urothelial Cells of Cattle Infected with Bovine

    • Authors: Francesca De Falco, Ivan Gentile, Pellegrino Cerino, Anna Cutarelli, Cornel Catoi, Sante Roperto
      First page: 621
      Abstract: Prohibitin 2 (PHB2), an inner mitochondrial membrane (IMM) protein, has recently been identified as a novel receptor involved in parkin-mediated mitophagy. In the field of veterinary medicine, the role of PHB2 in parkin-mediated mitophagy was described, for the first time, in urothelial cells of cattle, naturally infected with bovine papillomavirus (BPV). The BPV2 and BPV13 E5 oncoprotein, responsible for abortive infections in urothelial cells, was detected by RT-PCR. Severe ultrastructural abnormalities of the inner mitochondrial membrane were detected using transmission electron microscopy. PHB2 formed a functional complex with PHB1. PHB2 was significantly overexpressed in mitochondrial fractions from urothelial mucosa samples taken from cattle harbouring BPV infection. PHB2 overexpression could be attributed to mitochondrial dysfunction, as its expression levels in the cytosolic, microsomal, and nuclear fractions were seen to be unmodified. Immunoprecipitation studies revealed the interaction between PHB2 and phosphorylated forms of both PINK1 and parkin. Furthermore, PHB2 interacted with LC3-II, a marker of autophagosomal membranes and autophagy receptors, such as p62 and optineurin. PHB2 was shown to interact with transcription factor EB (TFEB), which is activated following parkin-mediated mitophagy, and embryonic stem cell-expressed Ras (ERAS), a constitutive protein coded by ERas. Western blot analysis revealed a significant overexpression of unphosphorylated TFEB in mitochondrial and nuclear fractions from urothelial mucosa samples from cattle suffering from BPV infection. Finally, PHB2 interacted with ERAS, believed to be involved in mitophagosome maturation. Taken together, the molecular and ultrastructural findings of this study suggested that BPV infection is responsible for parkin-dependent mitophagy, in the pathway of which PHB2 plays a crucial role.
      Citation: Pathogens
      PubDate: 2020-07-29
      DOI: 10.3390/pathogens9080621
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 622: Investigation of Macrolide Resistance
           Genotypes in Mycoplasma bovis Isolates from Canadian Feedlot Cattle

    • Authors: Andrea Kinnear, Tim A. McAllister, Rahat Zaheer, Matthew Waldner, Antonio C. Ruzzini, Sara Andrés-Lasheras, Sarah Parker, Janet E. Hill, Murray D. Jelinski
      First page: 622
      Abstract: Mycoplasma bovis is associated with bovine respiratory disease (BRD) and chronic pneumonia and polyarthritis syndrome (CPPS) in feedlot cattle. No efficacious vaccines for M. bovis exist; hence, macrolides are commonly used to control mycoplasmosis. Whole genome sequences of 126 M. bovis isolates, derived from 96 feedlot cattle over 12 production years, were determined. Antimicrobial susceptibility testing (AST) of five macrolides (gamithromycin, tildipirosin, tilmicosin, tulathromycin, tylosin) was conducted using a microbroth dilution method. The AST phenotypes were compared to the genotypes generated for 23S rRNA and the L4 and L22 ribosomal proteins. Mutations in domains II (nucleotide 748; E. coli numbering) and V (nucleotide 2059 and 2060) of the 23S rRNA (rrl) gene alleles were associated with resistance. All isolates with a single mutation at Δ748 were susceptible to tulathromycin, but resistant to tilmicosin and tildipirosin. Isolates with mutations in both domain II and V (Δ748Δ2059 or Δ748Δ2060) were resistant to all five macrolides. However, >99% of isolates were resistant to tildipirosin and tilmicosin, regardless of the number and positions of the mutations. Isolates with a Δ748 mutation in the 23S rRNA gene and mutations in L4 and L22 were resistant to all macrolides except for tulathromycin.
      Citation: Pathogens
      PubDate: 2020-07-30
      DOI: 10.3390/pathogens9080622
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 623: Detection of Epstein-Barr Virus DNA in
           Gastric Biopsies of Pediatric Patients with Dyspepsia

    • Authors: Abigail Morales-Sánchez, Javier Torres, María G. Cardenas-Mondragón, Carolina Romo-González, Margarita Camorlinga-Ponce, Lourdes Flores-Luna, Ezequiel M. Fuentes-Pananá
      First page: 623
      Abstract: In this study, we assessed the presence of Epstein-Barr virus (EBV) in gastric samples derived from pediatric patients with dyspeptic symptoms, aiming to understand whether EBV participates in the development of early gastric lesions influencing chronic inflammation, in conjunction with the Helicobacter pylori (Hp) bacterium. We analyzed EBV load in 236 gastric biopsies derived from 186 pediatric patients with chronic dyspepsia and compared it with EBV serology, Hp load and serology, and with immune cell infiltration. We found that 7.5% of patients were positive for EBV load, ranging from 240 to 29,685 genomic copies/μg of DNA. Hp genomic sequences were found in 24.7% of patients. EBV positive samples did not correlate with Hp status and were characterized by absent to moderate immune cell infiltration. To our knowledge, this is the first study addressing EBV load in the stomach in a large cohort of pediatric patients with dyspeptic symptoms, providing evidence of EBV localization in the gastric mucosa in early inflammatory lesions. The lack of correlation between EBV and both Hp infection and inflammation is perhaps explained by independent pathogenic mechanisms or because of the randomness of the gastritis sampling. This is also supported by a moderate association between EBV load and serology.
      Citation: Pathogens
      PubDate: 2020-07-30
      DOI: 10.3390/pathogens9080623
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 624: A Sensitive, One-Way Sequential Sieving
           Method to Isolate Helminths’ Eggs and Protozoal Oocysts from Lettuce for
           Genetic Identification

    • Authors: Annina R. Guggisberg, Cristian A. Alvarez Rojas, Philipp A. Kronenberg, Nadia Miranda, Peter Deplazes
      First page: 624
      Abstract: Different helminths and protozoa are transmitted to humans by oral uptake of environmentally resistant parasite stages after hand-to-mouth contact or by contaminated food and water. The aim of this study was to develop and validate a method for the simultaneous detection of parasite stages from fresh produce (lettuce) by a one-way isolation test kit followed by genetic identification (PCR, sequencing). Three sentinel zoonotic agents (eggs of Toxocara canis, Echinococcus multilocularis and oocysts of Toxoplasma gondii) were used to investigate the practicability and sensitivity of the method. The detection limits (100% positive results) in the recovery experiments were four Toxocara eggs, two E. multilocularis eggs and 18 T. gondii oocysts (in 4/5 replicates). In a field study, helminth DNA was detected in 14 of 157 lettuce samples including Hydatigera taeniaeformis (Syn. Taenia taeniaeformis) (four samples), T. polyacantha (three), T. martis (one), E. multilocularis (two) and Toxocara cati (four). Toxoplasma gondii was detected in six of 100 samples. In vivo testing in mice resulted in metacestode growth in all animals injected with 40–60 E. multilocularis eggs, while infection rates were 20–40% with 2–20 eggs. The developed diagnostic strategy is highly sensitive for the isolation and genetic characterisation of a broad range of parasite stages from lettuce, whereas the sensitivity of the viability tests needs further improvement.
      Citation: Pathogens
      PubDate: 2020-07-31
      DOI: 10.3390/pathogens9080624
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 625: The Prevalence of Chlamydia trachomatis and
           Three Other Non-Viral Sexually Transmitted Infections among Pregnant Women
           in Pemba Island Tanzania

    • Authors: Naomi C.A. Juliana, Saikat Deb, Sander Ouburg, Aishwarya Chauhan, Jolein Pleijster, Said M. Ali, Servaas A. Morré, Sunil Sazawal, Elena Ambrosino
      First page: 625
      Abstract: Efforts to map the burden of infections globally have shown a high prevalence of genital infections, including Chlamydia trachomatis, in sub-Saharan Africa. This retrospective study aimed to investigate the prevalence of selected non-viral genital infections among pregnant women in Pemba Island, Tanzania. Vaginal swabs were collected during pregnancy and stored in eNAT buffer. Detection of C. trachomatis, Neisseria gonorrheae, Trichomonas vaginalis, and Mycoplasma genitalium pathogens was performed by PCR using validated detection kits. Vaginal samples of 439 pregnant women between 16 and 48 years were tested. In fifty-five (12.5%) of them, at least one genital pathogen was detected. The most prevalent pathogen was T. vaginalis (7.1%), followed by C. trachomatis (4.6%) and M. genitalium (2.1%). None of the vaginal samples tested positive for N. gonorrheae. Consequently, among positive samples, 7.3% were for C. trachomatis and at least one other genital pathogen. This study provides insights on the burden of the four studied genital infections, and on the coinfections among pregnant women in Pemba Island, Tanzania. These results offer a starting point that can be useful to design further research in the field of maternal and child health in Pemba Island.
      Citation: Pathogens
      PubDate: 2020-07-31
      DOI: 10.3390/pathogens9080625
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 626: New Molecular Data on Filaria and its
           Wolbachia from Red Howler Monkeys (Alouatta macconnelli) in French
           Guiana—A Preliminary Study

    • Authors: Younes Laidoudi, Hacène Medkour, Anthony Levasseur, Bernard Davoust, Oleg Mediannikov
      First page: 626
      Abstract: Previous studies have reported filarial parasites of the genus Dipetalonema and Mansonella from French Guiana monkeys, based on morphological taxonomy. In this study, we screened blood samples from nine howler monkeys (Alouatta macconnelli) for the presence of filaria and Wolbachia DNA. The infection rates were 88.9% for filaria and 55.6% for wolbachiae. The molecular characterization, based on the 18S gene of filariids, revealed that A. macconnelli are infected with at least three species (Mansonella sp., Brugia sp. and an unidentified Onchocercidae species.). Since the 18S and cox1 generic primers are not very effective at resolving co-infections, we developed ITS genus-specific PCRs for Mansonella and Brugia genus. The results revealed coinfections in 75% of positives. The presence of Mansonella sp. and Brugia sp. was also confirmed by the 16S phylogenetic analysis of their associated Wolbachia. Mansonella sp., which close to the species from the subgenus Tetrapetalonema encountered in New World Monkeys, while Brugia sp. was identical to the strain circulating in French Guiana dogs. We propose a novel ITS1Brugia genus-specific qPCR. We applied it to screen for Brugia infection in howler monkeys and 66.7% were found to be positive. Our finding highlights the need for further studies to clarify the species diversity of neotropics monkeys by combining molecular and morphological features. The novel Brugia genus-specific qPCR assays could be an effective tool for the surveillance and characterization of this potential zoonosis.
      Citation: Pathogens
      PubDate: 2020-07-31
      DOI: 10.3390/pathogens9080626
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 627: Objective Olfactory Findings in Hospitalized
           Severe COVID-19 Patients

    • Authors: Jerome R. Lechien, Morgane Ducarme, Sammy Place, Carlos M. Chiesa-Estomba, Mohamad Khalife, Giacomo De Riu, Luigi Angelo Vaira, Christophe de Terwangne, Shahram Machayekhi, Arnaud Marchant, Fabrice Journe, Sven Saussez
      First page: 627
      Abstract: Objective: We investigate the prevalence of the self-reported and objective sudden loss of smell (SLS) in patients with severe coronavirus disease 2019 (COVID-19). Methods: Severe COVID-19 patients with self-reported SLS were recruited at hospitalization discharge. Epidemiological and clinical data were collected. The Sino-nasal Outcome Test-22 (SNOT-22) was used to evaluate rhinological complaints. Subjective olfactory and gustatory functions were assessed with the National Health and Nutrition Examination Survey (NHNES). Objective SLS was evaluated using psychophysical tests. Potential associations between olfactory evaluation and the clinical outcomes (duration of hospitalization; admission biology; one month serology (IgG), and chest computed tomography findings) were studied. Results: Forty-seven patients completed the study (25 females). Subjectively, eighteen (38.3%) individuals self-reported subjective partial or total SLS. Among them, only three and four were anosmic and hyposmic, respectively (38.9%). Considering the objective evaluation in the entire cohort, the prevalence of SLS was 21.3%. Elderly patients and those with diabetes had lower objective olfactory evaluation results than young and non-diabetic individuals. Conclusion: The prevalence of SLS in severe COVID-19 patients appears to be lower than previously estimated in mild-to-moderate COVID-19 forms. Future comparative studies are needed to explore the predictive value of SLS for COVID-19 severity.
      Citation: Pathogens
      PubDate: 2020-07-31
      DOI: 10.3390/pathogens9080627
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 628: Comparative Analysis of the Bacterial and
           Fungal Communities in the Gut and the Crop of Aedes albopictus Mosquitoes:
           A Preliminary Study

    • Authors: Morgane Guégan, Edwige Martin, Claire Valiente Moro
      First page: 628
      Abstract: The Asian tiger mosquito Aedes albopictus is a major pathogen vector and one of the world’s most invasive species. In recent years, the study of mosquito-associated microbiota has received growing interest for reducing transmission of mosquito-borne pathogens. Most of studies on mosquito microbiota mainly focused on the gut bacteria. However, microorganisms can also colonize other organs and are not restricted to bacteria. In mosquitoes, the crop is the primary storage organ for sugars from the nectar feeding before it is transferred into the midgut for digestion. No study has yet investigated whether this organ can harbor microorganisms in Ae. albopictus. By using high-throughput sequencing, this study is the first to describe the microbiota including both bacteria and fungi in sugar-fed Ae. albopictus males and females. The results showed the presence of diverse and rich bacterial and fungal communities in the crop of both sexes that did not strongly differ from the community composition and structure found in the gut. Altogether, our results provide a thorough description of the crop-associated microbiota in Ae. albopictus which can open new avenues for further studies on trophic interactions between the mosquito and its microbiota.
      Citation: Pathogens
      PubDate: 2020-08-01
      DOI: 10.3390/pathogens9080628
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 629: Primary Amebic Meningoencephalitis Related
           to Groundwater in Costa Rica: Diagnostic Confirmation of Three Cases and
           Environmental Investigation

    • Authors: Lissette Retana Moreira, Leidy Zamora Rojas, Muriel Grijalba Murillo, Silvia Elena Molina Castro, Elizabeth Abrahams Sandí
      First page: 629
      Abstract: During the first trimester of 2020, the Ministry of Health of Costa Rica reported the first three cases of primary amebic meningoencephalitis (PAM). In two cases, laboratory personnel of the hospitals preliminarily identified amoeboid forms in cerebrospinal fluid (CSF) samples. For the molecular confirmation of species, CSF samples were sent to our laboratory. We carried out microscopic analyses and exflagellation assays. Besides, samples were cultured in 2% casein hydrolysate medium and in non-nutrient agar plates supplemented with Escherichia coli. Finally, PCR and sequencing were employed for the molecular diagnosis and species identification. In all cases, the presence of Naegleria fowleri was confirmed. An environmental investigation to identify the possible infection sources was also performed. Water samples from hot springs and groundwater from an artisan well were collected and after filtration and culture in non-nutrient agar plates supplemented with E. coli, thermotolerance and exflagellation assays were carried out. For the positive samples, PCR and sequencing were performed, confirming the presence of N. fowleri in several water samples. The report of these cases and the possible association with hot springs has had a significant impact on the population and health authorities of Costa Rica.
      Citation: Pathogens
      PubDate: 2020-08-01
      DOI: 10.3390/pathogens9080629
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 630: Biofilm-Formation in Clonally Unrelated
           Multidrug-Resistant Acinetobacter baumannii Isolates

    • Authors: Aisha M. Alamri, Afnan A. Alsultan, Mohammad A. Ansari, Amani M. Alnimr
      First page: 630
      Abstract: This study analyzed the genotype, antibiotic resistance, and biofilm formation of Acinetobacter baumannii strains and assessed the correlation between biofilm formation, antibiotic resistance, and biofilm-related risk factors. A total of 207 non-replicate multi-drug-resistant A. baumannii strains were prospectively isolated. Phenotypic identification and antimicrobial susceptibility testing were carried out. Isolate biofilm formation ability was evaluated using the tissue culture plate (TCP), Congo red agar, and tube methods. Clonal relatedness between the strains was assessed by enterobacterial repetitive intergenic consensus-PCR genotyping. Of the 207 isolates, 52.5% originated from an intensive care unit setting, and pan resistance was observed against ceftazidime and cefepime, with elevated resistance (99–94%) to piperacillin/tazobactam, imipenem, levofloxacin, and ciprofloxacin. alongside high susceptibility to tigecycline (97.8%). The Tissue culture plate, Tube method, and Congo red agar methods revealed that 53.6%, 20.8%, and 2.7% of the strains were strong biofilm producers, respectively, while a significant correlation was observed between biofilm formation and device-originating respiratory isolates (p = 0.0009) and between biofilm formation in colonized vs. true infection isolates (p = 0.0001). No correlation was detected between antibiotic resistance and biofilm formation capacity, and the majority of isolates were clonally unrelated. These findings highlight the urgent need for implementing strict infection control measures in clinical settings.
      Citation: Pathogens
      PubDate: 2020-08-02
      DOI: 10.3390/pathogens9080630
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 631: Rickettsia africae an Agent of African Tick
           Bite Fever in Ticks Collected from Domestic Animals in Eastern Cape, South

    • Authors: Iweriebor, Nqoro, Obi
      First page: 631
      Abstract: Background: Ticks transmit a plethora of pathogens of zoonotic implications. Their distribution, diversity and the pathogens they transmit differ from one ecological location to another. Rickettsia africae is the agent of African tick bite fever found in South Africa, a zoonotic infection that is frequently reported among travelers who have visited many sub-Saharan African countries where the pathogen is prevalent. Methods: Ticks were collected from domestic animals in Raymond Nkandla Municipality, Eastern Cape, South Africa. The ticks were identified morphologically prior to DNA extraction followed by molecular identification of randomly selected ticks from the morphologically delineated groups. To assess for the presence of tick-borne pathogens belonging to Rickettsia spp. by PCR (polymerase chain reaction), we used specific primer pairs targeting the gltA, ompA and ompB genes. The selected amplified ticks, all positive ompB and forty three ompA amplicons were sequenced in a commercial sequencing facility. The obtained nucleotide sequences were edited and subjected to BLASTn for homology search and phylogenetic analyses were performed with MEGA 7 Version for genetic relationships with curated reference sequences in GenBank. Results: A total of 953 ticks collected in the study were delineated into three genera consisting of Amblyomma, Rhipicephalus and Hyalomma in decreasing order of abundance. The presence of rickettsial DNA was detected in 60/953 (6.3%) from the three genera of ticks screened. Genetic analyses of the DNA sequences obtained showed that they have phylogenetic relationship to members of the spotted fever group rickettsiae with R. africae, being the predominant SFGR (spotted fever group rickettsiae) detected in the screened ticks. Conclusion: This report shows that R. africae is the predominant spotted fever group rickettsiae in ticks collected from domestic animals in the study area and the human health impacts are not known.
      Citation: Pathogens
      PubDate: 2020-08-02
      DOI: 10.3390/pathogens9080631
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 632: The Clinical Infection with Pigeon
           Circovirus (PiCV) Leads to Lymphocyte B Apoptosis But Has No Effect on
           Lymphocyte T Subpopulation

    • Authors: Tomasz Stenzel, Daria Dziewulska, Bartłomiej Tykałowski, Andrzej Koncicki
      First page: 632
      Abstract: The pathology of pigeon circovirus (PiCV) is still unknown, but it is regarded as an immunosuppressant. This study aimed to find a correlation between PiCV natural infection and immunosuppression. The study was conducted with 56 pigeons divided into the following groups: PiCV-positive but showing (group S) or not (group I) non-specific clinical symptoms and asymptomatic pigeons negative for PiCV (group H). The percentage and apoptosis of T CD3+ and B IgM+ splenocytes; the expression of CD4, CD8, and IFN-γ genes in splenic mononuclear cells; the number of PiCV viral loads in the bursa of Fabricius; and the level of anti-PiCV antibodies were analyzed. The results showed that the percentage of B IgM+ cells was almost two-fold lower in group S than in group H, and that ca. 20% of the lymphocytes were apoptotic. No increased apoptosis was detected in TCD3+ subpopulation. The PiCV viral loads were approximately one thousand and ten thousand times higher in group S than in groups I and H, respectively. Our results indicate a possible correlation between the number of PiCV viral loads and severity of PiCV infection and confirm that PiCV infection leads to the suppression of humoral immunity by inducing B lymphocyte apoptosis.
      Citation: Pathogens
      PubDate: 2020-08-03
      DOI: 10.3390/pathogens9080632
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 633: Comparative Pathology of Pseudorabies in
           Different Naturally and Experimentally Infected Species—A Review

    • Authors: Julia Sehl, Jens Peter Teifke
      First page: 633
      Abstract: The pseudorabies virus (PRV) is an alphaherpesvirus and the causative agent of Aujeszky’s disease (AD). PRV infects a wide range of animal species including swine as the natural host as well as ruminants, carnivores, rodents and lagomorphs. In these species, except for the pig, PRV infection causes acute, severe disease, characterized by insatiable itching, and is always lethal. Horses, chickens and non-human primates have been shown to be largely resistant to PRV infection, while disease in humans is still controversial. PRV is a pantropic virus, which preferably invades neural tissue, but also infects epithelia of various organs, whereupon multisystemic lesions may result. Although AD is mainly associated with severe pruritus, also known as “mad itch”, there are notable differences regarding infection route, clinical signs, viral distribution and lesion patterns in different animal species. In this comprehensive review, we will present clinico-pathologic findings from different species, which have been either shown to be susceptible to PRV infection or have been tested experimentally.
      Citation: Pathogens
      PubDate: 2020-08-04
      DOI: 10.3390/pathogens9080633
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 634: Combining Computed Tomography and Histology
           Leads to an Evolutionary Concept of Hepatic Alveolar Echinococcosis

    • Authors: Johannes Grimm, Annika Beck, Juliane Nell, Julian Schmidberger, Andreas Hillenbrand, Ambros J. Beer, Balázs Dezsényi, Rong Shi, Meinrad Beer, Peter Kern, Doris Henne-Bruns, Wolfgang Kratzer, Peter Moller, Thomas FE Barth, Beate Gruener, Tilmann Graeter
      First page: 634
      Abstract: Alveolar echinococcosis (AE) is caused by the intermediate stage of Echinococcus multilocularis. We aimed to correlate computed tomography (CT) data with histology to identify distinct characteristics for different lesion types. We classified 45 samples into five types with the Echinococcus multilocularis Ulm Classification for Computed Tomography (EMUC-CT). The various CT lesions exhibited significantly different histological parameters, which led us to propose a progression model. The initial lesion fit the CT type IV classification, which comprises a single necrotic area with the central located laminated layer, a larger distance between laminated layer and border zone, a small fibrotic peripheral zone, and few small particles of Echinococcus multilocularis (spems). Lesions could progress through CT types I, II, and III, characterized by shorter distances between laminated layer and border zone, more spems inside and surrounding the lesion, and a pronounced fibrotic rim (mostly in type III). Alternatively, lesions could converge to a highly calcified, regressive state (type V). Our results suggest that the CT types mark sequential stages of the infection, which progress over time. These distinct histological patterns advance the understanding of interactions between AE and human host; moreover, they might become prognostically and therapeutically relevant.
      Citation: Pathogens
      PubDate: 2020-08-04
      DOI: 10.3390/pathogens9080634
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 635: Increased Risk of Acquisition of New Delhi
           Metallo-Beta-Lactamase-Producing Carbapenem-Resistant Enterobacterales
           (NDM-CRE) among a Cohort of COVID-19 Patients in a Teaching Hospital in
           Tuscany, Italy

    • Authors: Andrea Davide Porretta, Angelo Baggiani, Guglielmo Arzilli, Virginia Casigliani, Tommaso Mariotti, Francesco Mariottini, Giuditta Scardina, Daniele Sironi, Michele Totaro, Simona Barnini, Gaetano Pierpaolo Privitera
      First page: 635
      Abstract: We describe the epidemiology of New Delhi Metallo-Beta-Lactamase-Producing Carbapenem-Resistant Enterobacterales (NDM-CRE) colonization/infection in a cohort of COVID-19 patients in an Italian teaching hospital. These patients had an increased risk of NDM-CRE acquisition versus the usual patients (75.9 vs. 25.3 cases/10,000 patient days). The co-infection significantly increased the duration of hospital stay (32.9 vs. 15.8 days).
      Citation: Pathogens
      PubDate: 2020-08-05
      DOI: 10.3390/pathogens9080635
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 636: Development of an IPM Strategy for Thrips
           and Tomato spotted wilt virus in Processing Tomatoes in the Central Valley
           of California

    • Authors: Ozgur Batuman, Thomas A. Turini, Michelle LeStrange, Scott Stoddard, Gene Miyao, Brenna J. Aegerter, Li-Fang Chen, Neil McRoberts, Diane E. Ullman, Robert L. Gilbertson
      First page: 636
      Abstract: Tomato spotted wilt virus (TSWV; species Tomato spotted wilt orthotospovirus; genus Orthotospovirus; family Tospoviridae) is a thrips-transmitted virus that can cause substantial economic losses to many crops, including tomato (Solanum lycopersicum). Since 2005, TSWV emerged as an economically important virus of processing tomatoes in the Central Valley of California, in part due to increased populations of the primary thrips vector, western flower thrips (WFT; Frankliniella occidentalis). To develop an understanding of the epidemiology of TSWV in this region, population densities of WFT and incidence of TSWV were monitored in California’s processing tomato transplant-producing greenhouses and associated open fields from 2007 to 2013. Thrips were monitored with yellow sticky cards and in tomato flowers, whereas TSWV incidence was assessed with indicator plants and field surveys for virus symptoms. All thrips identified from processing tomato fields were WFT, and females were three-fold more abundant on sticky cards than males. Symptoms of TSWV infection were observed in all monitored processing tomato fields. Incidences of TSWV ranged from 1 to 20%, with highest incidence found in late-planted fields. There was no single primary inoculum source, and inoculum sources for thrips/TSWV varied depending on the production region. These results allowed us to develop a model for TSWV infection of processing tomatoes in the Central Valley of California. The model predicts that low levels of primary TSWV inoculum are amplified in early-planted tomatoes and other susceptible crops leading to highest levels of infection in later-planted fields, especially those with high thrips populations. Based upon these findings, an integrated pest management (IPM) strategy for TSWV in processing tomatoes in California was devised. This IPM strategy focuses on strategic field placement (identification of high-risk situations), planting TSWV- and thrips-free transplants, planting resistant varieties, monitoring for TSWV symptoms and thrips, roguing infected plants, thrips management targeting early generations, extensive sanitation after harvest, and strategic cropping to avoid overlap with winter bridge crops.
      Citation: Pathogens
      PubDate: 2020-08-05
      DOI: 10.3390/pathogens9080636
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 637: Field Experience of Antibody Testing against
           Mycoplasma bovis in Adult Cows in Commercial Danish Dairy Cattle Herds

    • Authors: Mette Bisgaard Petersen, Lars Pedersen, Lone Møller Pedersen, Liza Rosenbaum Nielsen
      First page: 637
      Abstract: Mycoplasma bovis in cattle is difficult to diagnose. Recently, the ID screen® mycoplasma bovis indirect ELISA (ID screen) was commercially released by IDVet. The objectives of this study were to: (1) gain and share experience of using the ID screen in adult dairy cows under field conditions; (2) determine the correlation between antibody levels in milk and serum and (3) compare the ID screen results with those of the Bio K 302 (BioX 302) ELISA from BioX Diagnostics. Paired serum and milk samples were collected from 270 cows from 12 Danish dairy herds with three categories of M. bovis disease history. The ID screen tested nearly all cows positive in all, but the three non-infected herds, while the BioX 302 tested very few cows positive. The ID screen is therefore a much more sensitive test than the BioX 302. However, cows in five exposed herds without signs of ongoing infection and two herds with no history of M. bovis infection also tested ID screen positive. Therefore, the performance and interpretation of the test must be investigated under field conditions in best practice test evaluation setups. A concordance correlation coefficient of 0.66 (95% CI: 0.59–0.72) between the ID screen serum and milk results indicates that milk samples can replace serum samples for the ID screen diagnosis of M. bovis in adult cows.
      Citation: Pathogens
      PubDate: 2020-08-06
      DOI: 10.3390/pathogens9080637
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 638: Characterization of Cutaneous Bacterial
           Microbiota from Superficial Pyoderma Forms in Atopic Dogs

    • Authors: Caitlin E. Older, Aline Rodrigues Hoffmann, Kathleen Hoover, Frane Banovic
      First page: 638
      Abstract: Although Staphylococcus pseudintermedius is considered the major pathogen associated with superficial canine pyoderma, no study has investigated the entire bacterial community in these lesions with molecular techniques. The objectives of this study were to characterize the bacterial microbiota in two forms of superficial canine pyoderma lesions, superficial bacterial folliculitis (SBF) and epidermal collarette (EC), especially in terms of the staphylococcal community. Swabs from 12 SBF and 9 EC lesions were obtained from eight and six atopic dogs, respectively. Eight samples from the axilla and groin of four healthy dogs served as controls. DNA was extracted for 16S rRNA gene sequencing and quantitative polymerase chain reaction of Staphylococcus spp. and S. pseudintermedius. Healthy skin samples harbored significantly more diverse bacterial communities than pyoderma samples. Healthy samples had communities that were more similar to each other, and were distinct from pyoderma samples. Staphylococcus spp. abundance was increased in pyoderma samples, especially those from EC samples. Although determining species-level identities of staphylococcal sequences revealed many species, S. pseudintermedius was the primary staphylococcal species found in all sample types. As expected, there are many differences in the microbiota when comparing healthy and canine pyoderma lesions samples. These lesions do not seem to be associated with a change in the relative abundance of specific Staphylococcus species, but simply an overall increase in Staphylococcus spp. abundance. The results of this study provide a starting point for future studies investigating how antimicrobial treatments may further change the microbiota associated with these lesions.
      Citation: Pathogens
      PubDate: 2020-08-06
      DOI: 10.3390/pathogens9080638
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 639: Factoring in the Complexity of the Cystic
           Fibrosis Lung to Understand Aspergillus fumigatus and Pseudomonas
           aeruginosa Interactions

    • Authors: Beswick, Amich, Gago
      First page: 639
      Abstract: Pseudomonas aeruginosa has long been established as the most prevalent respiratory pathogen in Cystic Fibrosis (CF) patients, with opportunistic infection causing profound morbidity and mortality. Recently, Aspergillus fumigatus has also been recognised as a key contributor to CF lung deterioration, being consistently associated with decreased lung function and worsened prognosis in these patients. As clinical evidence for the common occurrence of combined infection with these two pathogens increases, research into the mechanism and consequences of their interaction is becoming more relevant. Clinical evidence suggests a synergistic effect of combined infection, which translates into a poorer prognosis for the patients. In vitro results from the laboratory have identified a variety of possible synergistic and antagonistic interactions between A. fumigatus and P. aeruginosa. Here, we present a comprehensive overview of the complex environment of the CF lung and discuss how it needs to be considered to determine the exact molecular interactions that A. fumigatus and P. aeruginosa undergo during combined infection and their effects on the host.
      Citation: Pathogens
      PubDate: 2020-08-06
      DOI: 10.3390/pathogens9080639
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 640: Mycoplasma bovis
           Infections—Occurrence, Diagnosis and Control

    • Authors: Katarzyna Dudek, Robin A. J. Nicholas, Ewelina Szacawa, Dariusz Bednarek
      First page: 640
      Abstract: Mycoplasma bovis is a cause of bronchopneumonia, mastitis and arthritis but may also affect other main organs in cattle such us the eye, ear or brain. Despite its non-zoonotic character, M. bovis infections are responsible for substantial economic health and welfare problems worldwide. M. bovis has spread worldwide, including to countries for a long time considered free of the pathogen. Control of M. bovis infections is hampered by a lack of effective vaccines and treatments due to increasing trends in antimicrobial resistance. This review summarizes the latest data on the epizootic situation of M. bovis infections and new sources/routes of transmission of the infection, and discusses the progress in diagnostics. The review includes various recommendations and suggestions which could be applied to infection control programs.
      Citation: Pathogens
      PubDate: 2020-08-06
      DOI: 10.3390/pathogens9080640
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 641: Preclinical Models of Nontuberculous
           Mycobacteria Infection for Early Drug Discovery and Vaccine Research

    • Authors: Elisa Rampacci, Valentina Stefanetti, Fabrizio Passamonti, Marcela Henao-Tamayo
      First page: 641
      Abstract: Nontuberculous mycobacteria (NTM) represent an increasingly prevalent etiology of soft tissue infections in animals and humans. NTM are widely distributed in the environment and while, for the most part, they behave as saprophytic organisms, in certain situations, they can be pathogenic, so much so that the incidence of NTM infections has surpassed that of Mycobacterium tuberculosis in developed countries. As a result, a growing body of the literature has focused attention on the critical role that drug susceptibility tests and infection models play in the design of appropriate therapeutic strategies against NTM diseases. This paper is an overview of the in vitro and in vivo models of NTM infection employed in the preclinical phase for early drug discovery and vaccine development. It summarizes alternative methods, not fully explored, for the characterization of anti-mycobacterial compounds.
      Citation: Pathogens
      PubDate: 2020-08-06
      DOI: 10.3390/pathogens9080641
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 642: Development and Validation of a New TaqMan
           Real-Time PCR for Detection of ‘Candidatus Phytoplasma pruni’

    • Authors: Zala Kogej, Marina Dermastia, Nataša Mehle
      First page: 642
      Abstract: Phytoplasmas of the 16SrIII group are wide spread, and have a broad plant host range. Among these, ‘Candidatus phytoplasma pruni’ (‘Ca. P. pruni’; phytoplasmas of 16SrIII subgroup A) can cause serious diseases in Prunus species and ‘Ca. P. pruni’-related strains can infect other plant species, including grapevines. In this study, a new real-time PCR detection system was developed for ‘Ca. P. pruni’ using TaqMan chemistry. This test was designed to detect ‘Ca. P. pruni’, by amplifying the species-specific secY gene. In addition, a test to amplify the group-specific 16S rRNA gene region was also developed. The performances of both tests were evaluated. The test that amplifies the secY gene provided reliable and quick detection of ‘Ca. P. pruni’. Using the newly developed and validated test, ‘Ca. P. pruni’ was not found in any of the 434 field samples collected from different plants species grown in different regions of Slovenia.
      Citation: Pathogens
      PubDate: 2020-08-07
      DOI: 10.3390/pathogens9080642
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 643: Antimicrobial Resistance and Biofilm
           Formation Capacity of Salmonella enterica Serovar Enteritidis Strains
           Isolated from Poultry and Humans in Poland

    • Authors: Katarzyna Ćwiek, Kamila Korzekwa, Aleksandra Tabiś, Jacek Bania, Gabriela Bugla-Płoskońska, Alina Wieliczko
      First page: 643
      Abstract: Salmonella enterica ser. Enteritidis (S. enterica ser. Enteritidis) is the most frequently detected serovar in human salmonellosis, and its ability to produce a biofilm and the risk of transmission from animals and food of animal origin to humans are significant. The main aim of the present work was to compare S. enterica ser. Enteritidis strains isolated from poultry and human feces in terms of resistance profiles, prevalence of selected resistance genes, and their potential for biofilm formation, by assessing their biofilm growth intensity, the prevalence and expression of selected genes associated with this phenomenon, and the correlation between increased antimicrobial resistance and biofilm formation ability of the two tested groups of S. enterica ser. Enteritidis. This study showed a difference in antimicrobial resistance (minimal inhibitory concentration value) between S. enterica ser. Enteritidis groups; however, the majority of multidrug-resistant (MDR) strains were isolated from poultry (environmental samples from chicken broilers, turkey broilers, and laying hens). Differences in the prevalence of resistance genes were observed; the most common gene among poultry strains was floR, and that among strains from humans was blaTEM. S. enterica ser. Enteritidis strains isolated from poultry under the tested incubation conditions exhibited better biofilm growth than strains isolated from humans. A higher level of gene expression associated with the production of cellulose was only detected in the S48 strain isolated from poultry. On the other hand, increased expression of genes associated with quorum sensing was observed in two strains isolated from poultry farms and one strain isolated from human feces.
      Citation: Pathogens
      PubDate: 2020-08-07
      DOI: 10.3390/pathogens9080643
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 644: Analyses of the Root-Knot Nematode
           (Meloidogyne graminicola) Transcriptome during Host Infection Highlight
           Specific Gene Expression Profiling in Resistant Rice Plants

    • Authors: Anne-Sophie Petitot, Alexis Dereeper, Corinne Da Silva, Julie Guy, Diana Fernandez
      First page: 644
      Abstract: The plant-parasitic nematode Meloidogyne graminicola causes considerable damages to rice (Oryza sativa) culture. Resistance to M. graminicola in the related species Oryza glaberrima reduces root penetration by juveniles and stops further nematode development. M. graminicola genes expressed during O. sativa infection were previously characterized but no information is available about the molecular dialogue established with a resistant plant. We compared the M. graminicola transcriptomes of stage-two juveniles (J2s) before and during infection of susceptible or resistant rice. Among 36,121 M. graminicola genes surveyed, 367 were differentially expressed during infection of resistant or susceptible plants. Genes encoding cell wall-degrading enzymes, peptidases and neuropeptides were expressed for a longer time in resistant plants compared to susceptible plants. Conversely, genes related to nematode development were not activated in the resistant host. The majority of M. graminicola effector genes had similar expression patterns, whatever the host genotype. However, two venom allergen-like protein (VAP)-encoding genes were specifically induced in resistant plants and Mg-VAP1 silencing in J2s reduced their ability to colonize roots. This study highlighted that M. graminicola adapts its gene expression to the host susceptibility. Further investigation is required to assess the role of Mg-VAPs in the rice–nematode interaction.
      Citation: Pathogens
      PubDate: 2020-08-08
      DOI: 10.3390/pathogens9080644
      Issue No: Vol. 9, No. 8 (2020)
  • Pathogens, Vol. 9, Pages 545: Mycoplasma bovis in Spanish Cattle Herds:
           Two Groups of Multiresistant Isolates Predominate, with One Remaining
           Susceptible to Fluoroquinolones

    • Authors: Ana García-Galán, Laurent-Xavier Nouvel, Eric Baranowski, Ángel Gómez-Martín, Antonio Sánchez, Christine Citti, Christian de la Fe
      First page: 545
      Abstract: Mycoplasma bovis is an important bovine pathogen causing pneumonia, mastitis, and arthritis and is responsible for major economic losses worldwide. In the absence of an efficient vaccine, control of M. bovis infections mainly relies on antimicrobial treatments, but resistance is reported in an increasing number of countries. To address the situation in Spain, M. bovis was searched in 436 samples collected from beef and dairy cattle (2016–2019) and 28% were positive. Single-locus typing using polC sequences further revealed that two subtypes ST2 and ST3, circulate in Spain both in beef and dairy cattle, regardless of the regions or the clinical signs. Monitoring of ST2 and ST3 isolates in a minimum inhibitory concentration (MIC) to a panel of antimicrobials revealed one major difference when using fluoroquinolones (FQL): ST2 is more susceptible than ST3. Accordingly, whole-genome sequencing (WGS) further identified mutations in the gyrA and parC regions, encoding quinolone resistance-determining regions (QRDR) only in ST3 isolates. This situation shows the capacity of ST3 to accumulate mutations in QRDR and might reflect the selective pressure imposed by the extensive use of these antimicrobials. MIC values and detection of mutations by WGS also showed that most Spanish isolates are resistant to macrolides, lincosamides, and tetracyclines. Valnemulin was the only one effective, at least in vitro, against both STs.
      Citation: Pathogens
      PubDate: 2020-07-07
      DOI: 10.3390/pathogens9070545
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 546: SARS-CoV-2, ACE2, and Hydroxychloroquine:
           Cardiovascular Complications, Therapeutics, and Clinical Readouts in the
           Current Settings

    • Authors: Rajkumar Singh Kalra, Dhanendra Tomar, Avtar Singh Meena, Ramesh Kandimalla
      First page: 546
      Abstract: The rapidly evolving coronavirus disease 2019 (COVID-19, caused by severe acute respiratory syndrome coronavirus 2- SARS-CoV-2), has greatly burdened the global healthcare system and led it into crisis in several countries. Lack of targeted therapeutics led to the idea of repurposing broad-spectrum drugs for viral intervention. In vitro analyses of hydroxychloroquine (HCQ)’s anecdotal benefits prompted its widespread clinical repurposing globally. Reports of emerging cardiovascular complications due to its clinical prescription are revealing the crucial role of angiotensin-converting enzyme 2 (ACE2), which serves as a target receptor for SARS-CoV-2. In the present settings, a clear understanding of these targets, their functional aspects and physiological impact on cardiovascular function are critical. In an up-to-date format, we shed light on HCQ’s anecdotal function in stalling SARS-CoV-2 replication and immunomodulatory activities. While starting with the crucial role of ACE2, we here discuss the impact of HCQ on systemic cardiovascular function, its associated risks, and the scope of HCQ-based regimes in current clinical settings. Citing the extent of HCQ efficacy, the key considerations and recommendations for the use of HCQ in clinics are further discussed. Taken together, this review provides crucial insights into the role of ACE2 in SARS-CoV-2-led cardiovascular activity, and concurrently assesses the efficacy of HCQ in contemporary clinical settings.
      Citation: Pathogens
      PubDate: 2020-07-07
      DOI: 10.3390/pathogens9070546
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 547: Efficacy of Erymicin 200 Injections for
           Reducing Renibacterium salmoninarum and Controlling Vertical Transmission
           in an Inland Rainbow Trout Brood Stock

    • Authors: Eric R. Fetherman, Brad Neuschwanger, Tracy Davis, Colby L. Wells, April Kraft
      First page: 547
      Abstract: Bacterial Kidney Disease, caused by Renibacterium salmoninarum (Rs), is widespread and can cause significant mortality at most life stages in infected salmonids. Rs is commonly found in inland trout, which can be carriers of the bacterium. Lethal spawns can be used to control vertical transmission to progeny through the culling of eggs from infected parents, but can be costly, time-consuming, and can negatively impact important and rare brood stocks. Erymicin 200 is an Investigational New Animal Drug (INAD) intended to reduce Rs levels in hatchery brood stocks and control vertical transmission to progeny. We tested the efficacy of Erymicin 200 injections in a positive, hatchery-resident rainbow trout (Oncorhynchus mykiss) brood stock in Colorado, USA. Brood fish age two and three were injected with 25 mg per kg of body weight Erymicin 200 three times prior to spawning. Erymicin 200 was effective in reducing Rs to below detectable levels in treated fish. However, both negative treated and control brood fish produced positive progeny, suggesting that Erymicin 200 did not prevent the vertical transmission of Rs.
      Citation: Pathogens
      PubDate: 2020-07-07
      DOI: 10.3390/pathogens9070547
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 548: Spatial and Temporal Evolutionary Patterns
           in Puumala Orthohantavirus (PUUV) S Segment

    • Authors: Florian Binder, René Ryll, Stephan Drewes, Sandra Jagdmann, Daniela Reil, Melanie Hiltbrunner, Ulrike M. Rosenfeld, Christian Imholt, Jens Jacob, Gerald Heckel, Rainer G. Ulrich
      First page: 548
      Abstract: The S segment of bank vole (Clethrionomys glareolus)-associated Puumala orthohantavirus (PUUV) contains two overlapping open reading frames coding for the nucleocapsid (N) and a non-structural (NSs) protein. To identify the influence of bank vole population dynamics on PUUV S segment sequence evolution and test for spillover infections in sympatric rodent species, during 2010–2014, 883 bank voles, 357 yellow-necked mice (Apodemus flavicollis), 62 wood mice (A. sylvaticus), 149 common voles (Microtus arvalis) and 8 field voles (M. agrestis) were collected in Baden-Wuerttemberg and North Rhine-Westphalia, Germany. In total, 27.9% and 22.3% of bank voles were positive for PUUV-reactive antibodies and PUUV-specific RNA, respectively. One of eight field voles was PUUV RNA-positive, indicating a spillover infection, but none of the other species showed evidence of PUUV infection. Phylogenetic and isolation-by-distance analyses demonstrated a spatial clustering of PUUV S segment sequences. In the hantavirus outbreak years 2010 and 2012, PUUV RNA prevalence was higher in our study regions compared to non-outbreak years 2011, 2013 and 2014. NSs amino acid and nucleotide sequence types showed temporal and/or local variation, whereas the N protein was highly conserved in the NSs overlapping region and, to a lower rate, in the N alone coding part.
      Citation: Pathogens
      PubDate: 2020-07-08
      DOI: 10.3390/pathogens9070548
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 549: Methicillin-Resistant Staphylococcus aureus
           Diabetic Foot Crossed Infection: A Case Report

    • Authors: María Reina-Bueno, Inmaculada C. Palomo-Toucedo, Aurora Castro-Méndez, Gabriel Domínguez-Maldonado, María del Carmen Vázquez-Bautista
      First page: 549
      Abstract: This work presents a protocol to prevent the transmission of multidrug-resistant infections. We focus on the Diabetic Foot Unit Podiatry Clinic Area attached to the University of Seville in particular. The most common complication for patients with diabetes is leg ulcers. Together with neuropathy, vasculopathy, and immunological response disorder, these individuals have a high predisposition to developing infections. Staphylococcus aureus is a highly prevalent microorganism in humans which, at times, may act as a pathogen. Due mainly to indiscriminate abuse of antibiotics, the methicillin-resistant strain known by its initials as MRSA is the most extended nosocomial infection globally and is a severe community and hospital healthcare problem. This paper describes compliance with new general recommendations on cleaning, hygiene, and decontamination, in addition to implementation of this specific protocol, after detection of cross infection (healthcare-related infection) in the studied unit in two patients with MRSA-infected ulcers. After an in-depth bibliographical review, strict hand hygiene measures and use of non-sterile gloves were used when treating all patients with a diabetic foot. Finally, we reflect on the need to educate healthcare personnel to guarantee correct prescription of selected antibiotics. The role of the podiatrist in the multidisciplinary team is highlighted not only in terms of management and treatment of lesions in diabetic patients, but also as a healthcare agent for the detection and prevention of MRSA together with other multidrug-resistant infections.
      Citation: Pathogens
      PubDate: 2020-07-08
      DOI: 10.3390/pathogens9070549
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 550: Natural Compounds from the Marine Brown Alga
           Caulocystis cephalornithos with Potent In Vitro-Activity against the
           Parasitic Nematode Haemonchus contortus

    • Authors: Aya C. Taki, Robert Brkljača, Tao Wang, Anson V. Koehler, Guangxu Ma, Jill Danne, Sarah Ellis, Andreas Hofmann, Bill C. H. Chang, Abdul Jabbar, Sylvia Urban, Robin B. Gasser
      First page: 550
      Abstract: Eight secondary metabolites (1 to 8) were isolated from a marine sponge, a marine alga and three terrestrial plants collected in Australia and subsequently chemically characterised. Here, these natural product-derived compounds were screened for in vitro-anthelmintic activity against the larvae and adult stages of Haemonchus contortus (barber’s pole worm)—a highly pathogenic parasitic nematode of ruminants. Using an optimised, whole-organism screening system, compounds were tested on exsheathed third-stage larvae (xL3s) and fourth-stage larvae (L4s). Anthelmintic activity was initially evaluated on these stages based on the inhibition of motility, development and/or changes in morphology (phenotype). We identified two compounds, 6-undecylsalicylic acid (3) and 6-tridecylsalicylic acid (4) isolated from the marine brown alga, Caulocystis cephalornithos, with inhibitory effects on xL3 and L4 motility and larval development, and the induction of a “skinny-straight” phenotype. Subsequent testing showed that these two compounds had an acute nematocidal effect (within 1–12 h) on adult males and females of H. contortus. Ultrastructural analysis of adult worms treated with compound 4 revealed significant damage to subcuticular musculature and associated tissues and cellular organelles including mitochondria. In conclusion, the present study has discovered two algal compounds possessing acute anthelmintic effects and with potential for hit-to-lead progression. Future work should focus on undertaking a structure-activity relationship study and on elucidating the mode(s) of action of optimised compounds.
      Citation: Pathogens
      PubDate: 2020-07-09
      DOI: 10.3390/pathogens9070550
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 551: Allelic Variation and Selection in Effector
           Genes of Phytophthora infestans (Mont.) de Bary

    • Authors: Juan G. Morales, Astrid E. Gaviria, Elizabeth Gilchrist
      First page: 551
      Abstract: Phytophthora infestans is a devastating plant pathogen in several crops such as potato (Solanum tuberosum), tomato (Solanum lycopersicum) and Andean fruits such as tree tomato (Solanum betaceum), lulo (Solanum quitoense), uchuva (Physalis peruviana) and wild species in the genus Solanum sp. Despite intense research performed around the world, P. infestans populations from Colombia, South America, are poorly understood. Of particular importance is knowledge about pathogen effector proteins, which are responsible for virulence. The present work was performed with the objective to analyze gene sequences coding for effector proteins of P. infestans from isolates collected from different hosts and geographical regions. Several genetic parameters, phylogenetic analyses and neutrality tests for non-synonymous and synonymous substitutions were calculated. Non-synonymous substitutions were identified for all genes that exhibited polymorphisms at the DNA level. Significant negative selection values were found for two genes (PITG_08994 and PITG_12737) suggesting active coevolution with the corresponding host resistance proteins. Implications for pathogen virulence mechanisms and disease management are discussed.
      Citation: Pathogens
      PubDate: 2020-07-09
      DOI: 10.3390/pathogens9070551
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 552: Cyclodepsipeptide Biosynthesis in
           Hypocreales Fungi and Sequence Divergence of The Non-Ribosomal Peptide
           Synthase Genes

    • Authors: Urbaniak, Waśkiewicz, Trzebny, Koczyk, Stępień
      First page: 552
      Abstract: Fungi from the Hypocreales order synthesize a range of toxic non-ribosomal cyclic peptides with antimicrobial, insecticidal and cytotoxic activities. Entomopathogenic Beauveria, Isaria and Cordyceps as well as phytopathogenic Fusarium spp. are known producers of beauvericins (BEAs), beauvenniatins (BEAEs) or enniatins (ENNs). The compounds are synthesized by beauvericin/enniatin synthase (BEAS/ESYN1), which shows significant sequence divergence among Hypocreales members. We investigated ENN, BEA and BEAE production among entomopathogenic (Beauveria, Cordyceps, Isaria) and phytopathogenic (Fusarium) fungi; BEA and ENNs were quantified using an LC-MS/MS method. Phylogenetic analysis of partial sequences of putative BEAS/ESYN1 amplicons was also made. Nineteen fungal strains were identified based on sequence analysis of amplified ITS and tef-1α regions. BEA was produced by all investigated fungi, with F. proliferatum and F. concentricum being the most efficient producers. ENNs were synthesized mostly by F. acuminatum, F. avenaceum and C. confragosa. The phylogeny reconstruction suggests that ancestral BEA biosynthesis independently diverged into biosynthesis of other compounds. The divergent positioning of three Fusarium isolates raises the possibility of parallel acquisition of cyclic depsipeptide synthases in ancient complexes within Fusarium genus. Different fungi have independently evolved NRPS genes involved in depsipeptide biosynthesis, with functional adaptation towards biosynthesis of overlapping yet diversified metabolite profiles.
      Citation: Pathogens
      PubDate: 2020-07-09
      DOI: 10.3390/pathogens9070552
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 553: Non-Tuberculous Mycobacterial Diseases in

    • Authors: Aniello Meoli, Michela Deolmi, Rosanna Iannarella, Susanna Esposito
      First page: 553
      Abstract: Non-tuberculous mycobacteria (NTMs) are ubiquitous and opportunistic emerging bacteria with the potential to colonize and eventually infect either immunocompromised or immunocompetent individuals. In the last three decades, the prevalence of disease caused by NTMs has increased in several countries. The increased prevalence of NTM infection can be explained by an ageing population with rising comorbidities, HIV infection, the common use of immunosuppressive drugs, and improved diagnostic methods. The aim of this review is to demonstrate the clinical relevance of NTMs in children, describing their features and manifestations, diagnostic tools, and therapeutic approaches. We collected data from the literature about NTM infections in young patients over the past five years (2014–2019) using the keywords “non-tuberculous”, “mycobacteria”, “paediatric”, “NTM”, “cystic fibrosis”, and “children”. Recent literature points out that NTMs are ubiquitous, with several species including both those that are pathogens for humans and those that are not. This means that, if a mycobacterium is isolated from a patient’s specimen, we have to distinguish between a simple colonization and an NTM-related disease. The start of treatment depends on many factors that are necessary to consider, such as clinical and imaging features, patient comorbidity and immunocompetence, drug adverse effects, and compliance with a very long therapy that can last many months. Due to the increasing prevalence and clinical relevance of NTMs, guidelines for their optimal management, especially in the presence of chronic underlying disease, are urgently needed.
      Citation: Pathogens
      PubDate: 2020-07-09
      DOI: 10.3390/pathogens9070553
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 554: Clinical and Immunological Features of Human
           Leishmania (L.) infantum-Infection, Novel Insights Honduras, Central

    • Authors: Wilfredo Sosa-Ochoa, Concepción Zúniga, Luis Fernando Chaves, Gabriela Venicia Araujo Flores, Carmen Maria Sandoval Pacheco, Vania Lúcia Ribeiro da Matta, Carlos Eduardo Pereira Corbett, Fernando Tobias Silveira, Marcia Dalastra Laurenti
      First page: 554
      Abstract: Leishmania (Leishmania) infantum is the etiological agent of both American visceral leishmaniasis (AVL) and non-ulcerated cutaneous leishmaniasis (NUCL) in Honduras. Although AVL is the most severe clinical form of infection, recent studies have shown that human immune response to parasite infection can result in a clinical-immunological spectrum. The overall prevalence rate of infection and clinical-immunological profiles of the L. (L.) infantum infection in Amapala municipality, South Honduras was determined. We examined 576 individuals with diagnosis based on combined ELISA (IgG/IgM) and DTH assays. We also used genus-specific kDNA PCR and Hsp70 PCR-RFLP for NUCL cases. Clinical evaluation found 82% asymptomatic and 18% symptomatic individuals. All symptomatic cases (n = 104) showing NUCL were positive for parasites. We identified L. (L.) infantum species in 100% of the skin lesion scrapings and in 90% of the blood samples from NUCL cases studied. A total of 320 asymptomatic individuals were exposed (ELISA+ and/or DTH+), providing an overall L. (L.) infantum prevalence of 73.6%. Clinical, parasitological, and immunological evaluations suggest seven infection profiles, three asymptomatic and four symptomatic. This represents the first report on clinical and immunological features of human L. (L.) infantum-infection in Amapala municipality, Honduras.
      Citation: Pathogens
      PubDate: 2020-07-10
      DOI: 10.3390/pathogens9070554
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 555: An Investigation of the Effect of
           Catecholamines and Glucocorticoids on the Growth and Pathogenicity of
           Campylobacter jejuni

    • Authors: Brendha Truccollo, Paul Whyte, Declan J. Bolton
      First page: 555
      Abstract: Campylobacter spp. are major causes of foodborne illness globally, and are mostly transmitted through the consumption and handling of poultry. Campylobacter infections have widely variable outcomes, ranging from mild enteritis to severe illness, which are attributed to host interactions and the virulence of the infecting strain. In this study, in order to investigate the effect of host stress on the growth and pathogenicity of C. jejuni, three strains associated with human infection and two strains from broilers were subject to growth, motility, adhesion and invasion assays, in response to exposure to catecholamines; epinephrine, norepinephrine and the glucocorticoid neuroendocrine hormones corticosterone, cortisol and cortisone which are associated with stress in humans and broilers. Catecholamines resulted in significantly increased growth, adhesion and invasion of Caco-2 cells. Corticosterone promoted growth in one of five strains, and cortisone resulted in a significant increase in motility in two out of five strains, while no significant differences were observed with the addition of cortisol. It was concluded that stress-associated hormones, especially catecholamines, may promote growth and virulence in Campylobacter.
      Citation: Pathogens
      PubDate: 2020-07-10
      DOI: 10.3390/pathogens9070555
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 556: Prevalence and Risk Factors of Intestinal
           Parasite Infections in Greek Swine Farrow-To-Finish Farms

    • Authors: Isaia Symeonidou, Panagiotis Tassis, Athanasios Ι. Gelasakis, Eleni D. Tzika, Elias Papadopoulos
      First page: 556
      Abstract: Intestinal parasites, helminths, and protozoa challenge health and welfare of pigs and deteriorate the sustainability of swine farms leading to monetary losses. A multicentric survey was conducted for approximately one year. Overall, 1150 fecal samples were collected from eight intensive farms in Greece at regular intervals and examined by flotation and Ziehl-Neelsen techniques. Age, season, and time of last recorded antiparasitic treatment were assessed as possible risk factors using binary regression models. The overall prevalence of intestinal parasitism in pigs was 44.7%. The most frequently detected parasites in the studied population were the protozoa Balantidium coli (37.8%), followed by Entamoeba spp. (8.3%), Cystoisospora suis (6.0%), and the nematodes Ascaris suum (3.7%), Trichuris suis (2.5%), and Oesophagostomum spp. (1.4%). Distribution of intestinal parasites in different age groups was as expected. In autumn, the prevalence of Balantidium coli infection enhanced whereas the prevalence of Entamoeba spp. and Cystoisospora suis infections increased in spring. Time of last recorded antiparasitic treatment influenced Balantidium coli and Trichuris suis infection levels. Our results demonstrated that swine intestinal parasitism in intensive farms of Greece seems to be relatively common and highlighted the importance of proper laboratory examinations, as well as the need for tailored made control programs.
      Citation: Pathogens
      PubDate: 2020-07-10
      DOI: 10.3390/pathogens9070556
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 557: Immunogenicity of Non-Living Anthrax Vaccine
           Candidates in Cattle and Protective Efficacy of Immune Sera in A/J Mouse
           Model Compared to the Sterne Live Spore Vaccine

    • Authors: Jauro, Ndumnego, Ellis, Buys, Beyer, Heerden
      First page: 557
      Abstract: The Sterne live spore vaccine (SLSV, Bacillus anthracis strain 34F2) is the veterinary vaccine of choice against anthrax though contra-indicated for use with antimicrobials. However, the use of non-living anthrax vaccine (NLAV) candidates can overcome the SLSV limitation. In this study, cattle were vaccinated with either of the NLAV (purified recombinant PA (PrPA) or crude rPA (CrPA) and formaldehyde-inactivated spores (FIS of B. anthracis strain 34F2) and emulsigen-D®/alhydrogel® adjuvants) or SLSV. The immunogenicity of the NLAV and SLSV was assessed and the protective efficacies evaluated using a passive immunization mouse model. Polyclonal IgG (including the IgG1 subset) and IgM responses increased significantly across all vaccination groups after the first vaccination. Individual IgG subsets titres peaked significantly with all vaccines used after the second vaccination at week 5 and remained significant at week 12 when compared to week 0. The toxin neutralization (TNA) titres of the NLAV vaccinated cattle groups showed similar trends to those observed with the ELISA titres, except that the former were lower, but still significant, when compared to week 0. The opsonophagocytic assay indicated good antibody opsonizing responses with 75% (PrPA+FIS), 66% (CrPA+FIS) and 80% (SLSV) phagocytosis following spores opsonization. In the passive protection test, A/J mice transfused with purified IgG from cattle vaccinated with PrPA+FIS+Emulsigen-D®/Alhydrogel® and SLSV had 73% and 75% protection from challenge with B. anthracis strain 34F2 spores, respectively, whereas IgG from cattle vaccinated with CrPA+FIS+Emulsigen-D®/Alhydrogel® offered insignificant protection of 20%. There was no difference in protective immune response in cattle vaccinated twice with either the PrPA+FIS or SLSV. Moreover, PrPA+FIS did not show any residual side effects in vaccinated cattle. These results suggest that the immunogenicity and protective efficacy induced by the NLAV (PrPA+FIS) in the cattle and passive mouse protection test, respectively, are comparable to that induced by the standard SLSV.
      Citation: Pathogens
      PubDate: 2020-07-10
      DOI: 10.3390/pathogens9070557
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 558: Synthesis and Evaluation of Novel
           Ellipticines and Derivatives as Inhibitors of Phytophthora infestans

    • Authors: McKee, Zheng, O’Sullivan, Kehoe, Doyle Prestwich, Mackrill, McCarthy
      First page: 558
      Abstract: The pathogen Phytophthora infestans is responsible for worldwide catastrophic crop damage and discovery of new inhibitors of this organism is of paramount agricultural and industrial importance. Current strategies for crop treatment are inadequate with limitations of efficacy and market alternatives. Ellipticines have recently been reported to have fungicidal properties and have been assessed against P. infestans growth with promising results. We hereby report a probe of the ellipticine framework to investigate the alkyl subunit and screen a set ellipticines and derivatives to identify new lead compounds to act against P. infestans. A series of ellipticinium salt derivatives have been identified with exceptional growth inhibitory activity and apparent lack of toxicity towards a human cell-line surpassing the effect of known and marketed fungicides. This report identifies the potential of this natural product derivative as a novel fungicide.
      Citation: Pathogens
      PubDate: 2020-07-10
      DOI: 10.3390/pathogens9070558
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 559: Identification of A Putative T6SS Immunity
           Islet in Salmonella Typhi

    • Authors: Luke Barretto, Casey Fowler
      First page: 559
      Abstract: Typhoid fever is a major global health problem and is the result of systemic infections caused by the human-adapted bacterial pathogen Salmonella enterica serovar Typhi (S. Typhi). The pathology underlying S. Typhi infections significantly differ from infections caused by broad host range serovars of the same species, which are a common cause of gastroenteritis. Accordingly, identifying S. Typhi genetic factors that impart functionality absent from broad host range serovars offers insights into its unique biology. Here, we used an in-silico approach to explore the function of an uncharacterized 14-gene S. Typhi genomic islet. Our results indicated that this islet was specific to the S. enterica species, where it was encoded by the Typhi and Paratyphi A serovars, but was generally absent from non-typhoidal serovars. Evidence was gathered using comparative genomics and sequence analysis tools, and indicated that this islet was comprised of Type VI secretion system (T6SS) and contact-dependent growth inhibition (CDI) genes, the majority of which appeared to encode orphan immunity proteins that protected against the activities of effectors and toxins absent from the S. Typhi genome. We herein propose that this islet represents an immune system that protects S. Typhi against competing bacteria within the human gut.
      Citation: Pathogens
      PubDate: 2020-07-11
      DOI: 10.3390/pathogens9070559
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 560: Inflammatory Immune Responses and Gut
           Microbiota Changes Following Campylobacter coli Infection of IL-10-/- Mice
           with Chronic Colitis

    • Authors: Markus M. Heimesaat, Claudia Genger, Nina Biesemeier, Sigri Klove, Dennis Weschka, Soraya Mousavi, Stefan Bereswill
      First page: 560
      Abstract: Human infections with the food-borne enteropathogens Campylobacter are progressively rising. Recent evidence revealed that pre-existing intestinal inflammation facilitates enteropathogenic infection subsequently exacerbating the underlying disease. Given that only little is known about C. coli–host interactions and particularly during intestinal inflammation, the aim of the present study was to survey gastrointestinal colonization properties, gut microbiota changes and pro-inflammatory sequelae upon peroral C. coli-infection of IL-10-/- mice with chronic colitis. C. coli colonized the gastrointestinal tract of mice with varying efficiencies until day 28 post-infection and induced macroscopic and microscopic inflammatory changes as indicated by shorter colonic lengths, more distinct histopathological changes in the colonic mucosa and higher numbers of apoptotic colonic epithelial cells when compared to mock-infected controls. Furthermore, not only colonic innate and adaptive immune cell responses, but also enhanced systemic TNF-α secretion could be observed following C. coli as opposed to mock challenge. Notably, C. coli induced intestinal inflammatory sequelae were accompanied with gut microbiota shifts towards higher commensal enterobacterial loads in the infected gut lumen. Moreover, the pathogen translocated from the intestinal tract to extra-intestinal tissue sites in some cases, but never to systemic compartments. Hence, C. coli accelerates inflammatory immune responses in IL-10-/- mice with chronic colitis.
      Citation: Pathogens
      PubDate: 2020-07-11
      DOI: 10.3390/pathogens9070560
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 561: Parasitic Infections in African Humans and
           Non-Human Primates

    • Authors: Hacène Medkour, Inestin Amona, Younes Laidoudi, Bernard Davoust, Idir Bitam, Anthony Levasseur, Jean Akiana, Georges Diatta, Liliana Pacheco, Slim Gorsane, Cheikh Sokhna, Raquel Adriana Hernandez-Aguilar, Amanda Barciela, Florence Fenollar, Didier Raoult, Oleg Mediannikov
      First page: 561
      Abstract: Different protozoa and metazoa have been detected in great apes, monkeys and humans with possible interspecies exchanges. Some are either nonpathogenic or their detrimental effects on the host are not yet known. Others lead to serious diseases that can even be fatal. Their survey remains of great importance for public health and animal conservation. Fecal samples from gorillas (Gorilla gorilla) and humans living in same area in the Republic of Congo, chimpanzees (Pan troglodytes) from Senegal and one other from the Republic of Congo, Guinea baboons (Papio papio) from Senegal, hamadryas baboons (Papio hamadryas) from Djibouti and Barbary macaques (Macaca sylvanus) from Algeria, were collected. DNA was extracted and screened using specific qPCR assays for the presence of a large number of helminths and protozoa. Positive samples were then amplified in standard PCRs and sequenced when possible. Overall, infection rate was 36.5% in all non-human primates (NHPs) and 31.6% in humans. Great apes were more often infected (63.6%) than monkeys (7.3%). At least twelve parasite species, including ten nematodes and two protozoa were discovered in NHPs and five species, including four nematodes and a protozoan in humans. The prevalences of Giarida lamblia, Necator americanus, Enterobius vermicularis, Strongyloides stercoralis were similar between gorillas and human community co-habiting the same forest ecosystem in the Republic of Congo. In addition, human specific Mansonella perstans (5.1%) and other Mansonella spp. (5.1%) detected in these gorillas suggest a possible cross-species exchange. Low prevalence (2%) of Ascaris lumbricoides, Enterobius vermicularis, Strongyloides stercoralis were observed in chimpanzees, as well as a high prevalence of Abbreviata caucasica (57.1%), which should be considered carefully as this parasite can affect other NHPs, animals and humans. The Barbary macaques were less infected (7.2%) and Oesophagostomum muntiacum was the main parasite detected (5.8%). Finally, we report the presence of Pelodera sp. and an environmental Nematoda DNAs in chimpanzee feces, Nematoda sp. and Bodo sp. in gorillas, as well as DNA of uncharacterized Nematoda in apes and humans, but with a relatively lower prevalence in humans. Prevalence of extraintestinal parasites remains underestimated since feces are not the suitable sampling methods. Using non-invasive sampling (feces) we provide important information on helminths and protozoa that can infect African NHPs and human communities living around them. Public health and animal conservation authorities need to be aware of these infections, as parasites detected in African NHPs could affect both human and other animals’ health.
      Citation: Pathogens
      PubDate: 2020-07-11
      DOI: 10.3390/pathogens9070561
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 562: Molecular Characterization of Echinococcus
           granulosus sensu lato from Humans in Slovenia

    • Authors: Barbara Šoba, Špela Gašperšič, Darja Keše, Tadeja Kotar
      First page: 562
      Abstract: The larval form of tapeworms of the Echinococcus granulosus sensu lato species cluster cause an important zoonotic infection, cystic echinococcosis (CE). Molecular characterization of the cluster’s isolates from different hosts greatly contributes to a better understanding of its transmission dynamics. To date, no genetic information is available on CE in Slovenia. In this work, we characterized isolates from human CE cases. Parasite samples from 18 patients were collected, together with the patients’ demographic and clinical data. Genomic DNA was analyzed by conventional PCR and sequencing at four mitochondrial loci (cytochrome c oxidase subunit 1, cox1; NADH dehydrogenase subunit 1, nad1; NADH dehydrogenase subunit 5, nad5; and small ribosomal RNA, rrnS). Thirteen isolates were successfully amplified and sequenced. Seven (58.8%) patients were infected with E. granulosus sensu stricto (s.s.) G1, five (38.5%) with E. canadensis G7 and one (7.7%) with E. granulosus s.s. G3. Echinococcus canadensis G7, the pig genotype, was identified exclusively in autochthonous Slovenes, while the patients originating from the Western Balkans were all infected with E. granulosus s.s. Our findings suggest that pigs are important intermediate hosts for human CE in Slovenia.
      Citation: Pathogens
      PubDate: 2020-07-12
      DOI: 10.3390/pathogens9070562
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 563: Aujeszky’s Disease and the Development of
           the Marker/DIVA Vaccination Concept

    • Authors: Thomas C. Mettenleiter
      First page: 563
      Abstract: Aujeszky’s disease or pseudorabies is an infection of animals caused by Suid alphaherpesvirus 1, also designated as pseudorabies virus (PrV). Whereas many mammals are susceptible to PrV, only pigs are able to survive productive infection. Early reports on this disease originate from cattle and companion animals with the hallmark sign of “mad itch”, meaning development of pruritus. Although first reports date back to the early 19th century, it was Aladár Aujeszky who in 1902 described this disease, which has since been named after him, as a separate entity. AD expanded in the 20th century, despite efforts to control this infection in the growing pig farming industry. Live-attenuated vaccines were developed in the early 1960s, which assisted early eradication efforts. A major breakthrough in animal vaccinology occurred in the mid-1980s, when it was found that several live-attenuated PrV vaccine strains lacked a significant portion of the genome, including the gene encoding a major immunogenic viral envelope glycoprotein. Upon the development of a suitable serological assay, the first marker vaccine/DIVA concept (differentiating infected from vaccinated animals) was developed. Moreover, the first genetically modified live vaccines emanated from molecular work on PrV. Thus, AD serves as a hallmark for the history of veterinary virology as well as for pioneering novel strategies for controlling animal infectious diseases.
      Citation: Pathogens
      PubDate: 2020-07-12
      DOI: 10.3390/pathogens9070563
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 564: Mosquito Mycobiota: An Overview of
           Non-Entomopathogenic Fungal Interactions

    • Authors: Simon Malassigné, Claire Valiente Moro, Patricia Luis
      First page: 564
      Abstract: The growing expansion of mosquito vectors leads to the emergence of vector-borne diseases in new geographic areas and causes major public health concerns. In the absence of effective preventive treatments against most pathogens transmitted, vector control remains one of the most suitable strategies to prevent mosquito-borne diseases. Insecticide overuse raises mosquito resistance and deleterious impacts on the environment and non-target species. Growing knowledge of mosquito biology has allowed the development of alternative control methods. Following the concept of holobiont, mosquito-microbiota interactions play an important role in mosquito biology. Associated microbiota is known to influence many aspects of mosquito biology such as development, survival, immunity or even vector competence. Mosquito-associated microbiota is composed of bacteria, fungi, protists, viruses and nematodes. While an increasing number of studies have focused on bacteria, other microbial partners like fungi have been largely neglected despite their huge diversity. A better knowledge of mosquito-mycobiota interactions offers new opportunities to develop innovative mosquito control strategies. Here, we review the recent advances concerning the impact of mosquito-associated fungi, and particularly nonpathogenic fungi, on life-history traits (development, survival, reproduction), vector competence and behavior of mosquitoes by focusing on Culex, Aedes and Anopheles species.
      Citation: Pathogens
      PubDate: 2020-07-12
      DOI: 10.3390/pathogens9070564
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 565: Mutational Frequencies of SARS-CoV-2 Genome
           during the Beginning Months of the Outbreak in USA

    • Authors: Neha Kaushal, Yogita Gupta, Mehendi Goyal, Svetlana F. Khaiboullina, Manoj Baranwal, Subhash C. Verma
      First page: 565
      Abstract: SARS-CoV-2 has spread very quickly from its first reported case on 19 January 2020 in the United Stated of America, leading WHO to declare pandemic by 11 March 2020. RNA viruses accumulate mutations following replication and passage in human population, which prompted us to determine the rate and the regions (hotspots) of the viral genome with high rates of mutation. We analyzed the rate of mutation accumulation over a period of 11 weeks (submitted between 19th January to 15 April 2020) in USA SARS-CoV-2 genome. Our analysis identified that majority of the viral genes accumulated mutations, although with varying rates and these included NSP2, NSP3, RdRp, helicase, Spike, ORF3a, ORF8, and Nucleocapsid protein. Sixteen mutations accumulated in Spike protein in which four mutations are located in the receptor binding domain. Intriguingly, we identified a fair number of viral proteins (NSP7, NSP9, NSP10, NSP11, Envelop, ORF6, and ORF7b proteins), which did not accumulate any mutation. Limited changes in these proteins may suggest that they have conserved functions, which are essential for virus propagation. This provides a basis for a better understanding of the genetic variation in SARS-CoV-2 circulating in the US, which could help in identifying potential therapeutic targets for controlling COVID-19.
      Citation: Pathogens
      PubDate: 2020-07-13
      DOI: 10.3390/pathogens9070565
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 566: Biotic Factors Influence Microbiota of Nymph
           Ticks from Vegetation in Sydney, Australia

    • Authors: Shona Chandra, Jan Šlapeta
      First page: 566
      Abstract: Ticks are haematophagous ectoparasites of medical and veterinary significance due to their excellent vector capacity. Modern sequencing techniques enabled the rapid sequencing of bacterial pathogens and symbionts. This study’s aims were two-fold; to determine the nymph diversity in Sydney, and to determine whether external biotic factors affect the microbiota. Tick DNA was isolated, and the molecular identity was determined for nymphs at the cox1 level. The tick DNA was subjected to high throughput DNA sequencing to determine the bacterial profile and the impact of biotic factors on the microbiota. Four nymph tick species were recovered from Sydney, NSW: Haemaphysalis bancrofti, Ixodes holocyclus, Ixodes trichosuri and Ixodes tasmani. Biotic factors, notably tick species and geography, were found to have a significance influence on the microbiota. The microbial analyses revealed that Sydney ticks display a core microbiota. The dominating endosymbionts among all tick species were Candidatus Midichloria sp. Ixholo1 and Candidatus Midichloria sp. Ixholo2. A novel Candidatus Midichloria sp. OTU_2090 was only found in I. holocyclus ticks (nymph: 96.3%, adult: 75.6%). Candidatus Neoehrlichia australis and Candidatus Neoehrlichia arcana was recovered from I. holocyclus and one I. trichosuri nymph ticks. Borrelia spp. was absent from all ticks. This study has shown that nymph and adult ticks carry different bacteria, and a tick bite in Sydney, Australia will result in different bacterial transfer depending on tick life stage, tick species and geography.
      Citation: Pathogens
      PubDate: 2020-07-13
      DOI: 10.3390/pathogens9070566
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 567: Legionella Diversity and Spatiotemporal
           Variation in the Occurrence of Opportunistic Pathogens within a Large
           Building Water System

    • Authors: Helen Y. Buse, Brian J. Morris, Vicente Gomez-Alvarez, Jeffrey G. Szabo, John S. Hall
      First page: 567
      Abstract: Understanding Legionella survival mechanisms within building water systems (BWSs) is challenging due to varying engineering, operational, and water quality characteristics unique to each system. This study aimed to evaluate Legionella, mycobacteria, and free-living amoebae occurrence within a BWS over 18–28 months at six locations differing in plumbing material and potable water age, quality, and usage. A total of 114 bulk water and 57 biofilm samples were analyzed. Legionella culturability fluctuated seasonally with most culture-positive samples being collected during the winter compared to the spring, summer, and fall months. Positive and negative correlations between Legionella and L. pneumophila occurrence and other physiochemical and microbial water quality parameters varied between location and sample types. Whole genome sequencing of 19 presumptive Legionella isolates, from four locations across three time points, identified nine isolates as L. pneumophila serogroup (sg) 1 sequence-type (ST) 1; three as L. pneumophila sg5 ST1950 and ST2037; six as L. feeleii; and one as Ochrobactrum. Results showed the presence of a diverse Legionella population with consistent and sporadic occurrence at four and two locations, respectively. Viewed collectively with similar studies, this information will enable a better understanding of the engineering, operational, and water quality parameters supporting Legionella growth within BWSs.
      Citation: Pathogens
      PubDate: 2020-07-13
      DOI: 10.3390/pathogens9070567
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 568: Seroprevalence of Hepatitis E Virus in
           Forestry Workers from Trentino-Alto Adige Region (Northern Italy)

    • Authors: Marina Monini, Fabio Ostanello, Alessandra Dominicis, Valentina Tagliapietra, Gabriele Vaccari, Annapaola Rizzoli, Claudia M. Trombetta, Emanuele Montomoli, Ilaria Di Bartolo
      First page: 568
      Abstract: People with some occupational or recreational activities, such as hunters and veterinarians, may have increased risk to be infected by the hepatitis E virus (HEV). The aim of the present study was to establish whether forestry workers could be considered at a higher risk of HEV infection than a control group. One hundred and fifty sera from forestry workers and a control group of 85 sera were analysed by anti-HEV IgG antibodies detection using a commercial ELISA kit. The anti-HEV IgG seroprevalence was 14% for forestry workers and 9.4% for the control group. Comparing the risk of HEV infection in the two groups, there was no difference in the odds ratio. However, the seroprevalence in older subjects was higher in the forestry workers than in the control group. Two sera from forestry workers were also positive for anti-HEV IgM, and, in one of them, HEV-RNA was detected. Our findings showed an increase of seroprevalence with age, which is likely to reflect cumulative exposure to HEV over time. The occupation of forestry workers did not seem to be associated with a higher risk of HEV infection. The study provided new insights into the risk of acquiring HEV in occupational exposure workers with open-air activities.
      Citation: Pathogens
      PubDate: 2020-07-14
      DOI: 10.3390/pathogens9070568
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 569: Microbial Etiology and Prevention of Dental
           Caries: Exploiting Natural Products to Inhibit Cariogenic Biofilms

    • Authors: Xiuqin Chen, Eric Banan-Mwine Daliri, Namhyeon Kim, Jong-Rae Kim, Daesang Yoo, Deog-Hwan Oh
      First page: 569
      Abstract: Dental caries is one of the most common microbe-mediated oral diseases in human beings. At present, the accepted etiology of caries is based on a four-factor theory that includes oral microorganisms, oral environment, host, and time. Excessive exposure to dietary carbohydrates leads to the accumulation of acid-producing and acid-resistant microorganisms in the mouth. Dental caries is driven by dysbiosis of the dental biofilm adherent to the enamel surface. Effective preventive methods include inhibiting the cariogenic microorganisms, treatment with an anti-biofilm agent, and sugar intake control. The goal is to reduce the total amount of biofilm or the levels of specific pathogens. Natural products could be recommended for preventing dental caries, since they may possess fewer side effects in comparison with synthetic antimicrobials. Herein, the mechanisms of oral microbial community development and functional specialization are discussed. We highlight the application of widely explored natural products in the last five years for their ability to inhibit cariogenic microorganisms.
      Citation: Pathogens
      PubDate: 2020-07-14
      DOI: 10.3390/pathogens9070569
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 570: Phylogenetic Analyses of Rotavirus A from
           Cattle in Uruguay Reveal the Circulation of Common and Uncommon Genotypes
           and Suggest Interspecies Transmission

    • Authors: Matías Castells, Rubén Darío Caffarena, María Laura Casaux, Carlos Schild, Samuel Miño, Felipe Castells, Daniel Castells, Matías Victoria, Franklin Riet-Correa, Federico Giannitti, Viviana Parreño, Rodney Colina
      First page: 570
      Abstract: Uruguay is one of the main exporters of beef and dairy products, and cattle production is one of the main economic sectors in this country. Rotavirus A (RVA) is the main pathogen associated with neonatal calf diarrhea (NCD), a syndrome that leads to significant economic losses to the livestock industry. The aims of this study are to determine the frequency of RVA infections, and to analyze the genetic diversity of RVA strains in calves in Uruguay. A total of 833 samples from dairy and beef calves were analyzed through RT-qPCR and sequencing. RVA was detected in 57.0% of the samples. The frequency of detection was significantly higher in dairy (59.5%) than beef (28.4%) calves (p < 0.001), while it did not differ significantly among calves born in herds that were vaccinated (64.0%) or not vaccinated (66.7%) against NCD. The frequency of RVA detection and the viral load were significantly higher in samples from diarrheic (72.1%, 7.99 log10 genome copies/mL of feces) than non-diarrheic (59.9%, 7.35 log10 genome copies/mL of feces) calves (p < 0.005 and p = 0.007, respectively). The observed G-types (VP7) were G6 (77.6%), G10 (20.7%), and G24 (1.7%), while the P-types were P[5] (28.4%), P[11] (70.7%), and P[33] (0.9%). The G-type and P-type combinations were G6P[11] (40.4%), G6P[5] (38.6%), G10P[11] (19.3%), and the uncommon genotype G24P[33] (1.8%). VP6 and NSP1-5 genotyping were performed to better characterize some strains. The phylogenetic analyses suggested interspecies transmission, including transmission between animals and humans.
      Citation: Pathogens
      PubDate: 2020-07-14
      DOI: 10.3390/pathogens9070570
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 571: Intracellular Growth and Cell Cycle
           Progression are Dependent on (p)ppGpp Synthetase/Hydrolase in Brucella

    • Authors: Mathilde Van der Henst, Elodie Carlier, Xavier De Bolle
      First page: 571
      Abstract: Brucella abortus is a pathogenic bacterium able to proliferate inside host cells. During the first steps of its trafficking, it is able to block the progression of its cell cycle, remaining at the G1 stage for several hours, before it reaches its replication niche. We hypothesized that starvation mediated by guanosine tetra- or penta-phosphate, (p)ppGpp, could be involved in the cell cycle arrest. Rsh is the (p)ppGpp synthetase/hydrolase. A B. abortus ∆rsh mutant is unable to grow in minimal medium, it is unable to survive in stationary phase in rich medium and it is unable to proliferate inside RAW 264.7 macrophages. A strain producing the heterologous constitutive (p)ppGpp hydrolase Mesh1b is also unable to proliferate inside these macrophages. Altogether, these data suggest that (p)ppGpp is necessary to allow B. abortus to adapt to its intracellular growth conditions. The deletion of dksA, proposed to mediate a part of the effect of (p)ppGpp on transcription, does not affect B. abortus growth in culture or inside macrophages. Expression of a gene coding for a constitutively active (p)ppGpp synthetase slows down growth in rich medium and inside macrophages. Using an mCherry–ParB fusion able to bind to the replication origin of the main chromosome of B. abortus, we observed that expression of the constitutive (p)ppGpp synthetase gene generates an accumulation of bacteria at the G1 phase. We thus propose that (p)ppGpp accumulation could be one of the factors contributing to the G1 arrest observed for B. abortus in RAW 264.7 macrophages.
      Citation: Pathogens
      PubDate: 2020-07-14
      DOI: 10.3390/pathogens9070571
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 572: Frequent Recombination Events in Leishmania
           donovani: Mining Population Data

    • Authors: Igor B. Rogozin, Arzuv Charyyeva, Ivan A. Sidorenko, Vladimir N. Babenko, Vyacheslav Yurchenko
      First page: 572
      Abstract: The Leishmania donovani species complex consists of all L. donovani and L. infantum strains mainly responsible for visceral leishmaniasis (VL). It was suggested that genome rearrangements in Leishmania spp. occur very often, thus enabling parasites to adapt to the different environmental conditions. Some of these rearrangements may be directly linked to the virulence or explain the reduced efficacy of antimonial drugs in some isolates. In the current study, we focused on a large-scale analysis of putative gene conversion events using publicly available datasets. Previous population study of L. donovani suggested that population variability of L. donovani is relatively low, however the authors used masking procedures and strict read selection criteria. We decided to re-analyze DNA-seq data without masking sequences, because we were interested in the most dynamic fraction of the genome. The majority of samples have an excess of putative gene conversion/recombination events in the noncoding regions, however we found an overall excess of putative intrachromosomal gene conversion/recombination in the protein coding genes, compared to putative interchromosomal gene conversion/recombination events.
      Citation: Pathogens
      PubDate: 2020-07-15
      DOI: 10.3390/pathogens9070572
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 573: Leptospira Infections in Domestic and Wild

    • Authors: Giovanni Cilia, Fabrizio Bertelloni, Filippo Fratini
      First page: 573
      Abstract: Leptospirosis is a worldwide-distributed, re-emerging zoonosis due to the large variety of wild and domestic animal species that can play the role of natural or accidental host. Currently, specific animal species play an important role as the reservoir for particular Leptospira serovars, although recent investigations have highlighted new host–pathogen interactions involved in Leptospira epidemiology. Furthermore, the constant modification of ecosystems and wildlife habitats and the constantly increasing number of animal species moving towards urban or peri-urban areas are increasing the possibility of direct or indirect contacts between wildlife and domestic animals; furthermore, the constant modification of animal leptospirosis also causes problems for human health. The studies published in this Special Issue have evidenced and confirmed the hidden role of a large variety of animal species, domestic and wild, in the leptospirosis epidemiology. They highlighted the necessity for continuous monitoring and large-scale surveillance studies to better understand this neglected and re-emerging zoonosis.
      Citation: Pathogens
      PubDate: 2020-07-15
      DOI: 10.3390/pathogens9070573
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 574: Erratum: Fukuyama, K., et al. Evaluation of
           the Immunomodulatory Ability of Lactic Acid Bacteria Isolated from Feedlot
           Cattle Against Mastitis Using a Bovine Mammary Epithelial Cells In Vitro
           Assay. Pathogens 2020, 9, 410

    • Authors: Kohtaro Fukuyama, Md. Aminul Islam, Michihiro Takagi, Wakako Ikeda-Ohtsubo, Shoichiro Kurata, Hisashi Aso, Maria Elena Fatima Nader-Macías, Graciela Vignolo, Julio Villena, Haruki Kitazawa
      First page: 574
      Abstract: The authors would like to make the following corrections about the published paper [...]
      Citation: Pathogens
      PubDate: 2020-07-16
      DOI: 10.3390/pathogens9070574
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 575: Vector Competence of Aedes aegypti, Aedes
           albopictus and Culex quinquefasciatus from Brazil and New Caledonia for
           Three Zika Virus Lineages

    • Authors: Rosilainy S. Fernandes, Olivia O’Connor, Maria Ignez L. Bersot, Dominique Girault, Marguerite R. Dokunengo, Nicolas Pocquet, Myrielle Dupont-Rouzeyrol, Ricardo Lourenço-de-Oliveira
      First page: 575
      Abstract: Zika virus (ZIKV) has caused severe epidemics in South America beginning in 2015, following its spread through the Pacific. We comparatively assessed the vector competence of ten populations of Aedes aegypti and Ae. albopictus from Brazil and two of Ae. aegypti and one of Culex quinquefasciatus from New Caledonia to transmit three ZIKV isolates belonging to African, Asian and American lineages. Recently colonized mosquitoes from eight distinct sites from both countries were orally challenged with the same viral load (107 TCID50/mL) and examined after 7, 14 and 21 days. Cx. quinquefasciatus was refractory to infection with all virus strains. In contrast, although competence varied with geographical origin, Brazilian and New Caledonian Ae. aegypti could transmit the three ZIKV lineages, with a strong advantage for the African lineage (the only one reaching saliva one-week after challenge). Brazilian Ae. albopictus populations were less competent than Ae. aegypti populations. Ae. albopictus generally exhibited almost no transmission for Asian and American lineages, but was efficient in transmitting the African ZIKV. Viral surveillance and mosquito control measures must be strengthened to avoid the spread of new ZIKV lineages and minimize the transmission of viruses currently circulating.
      Citation: Pathogens
      PubDate: 2020-07-16
      DOI: 10.3390/pathogens9070575
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 576: Molecular Detection and Genetic Diversity of
           Toxoplasma gondii Oocysts in Cat Faeces from Klang Valley, Malaysia, Using
           B1 and REP Genes in 2018

    • Authors: Mohammed Nasiru Wana, Mohamad Aris Mohd Moklas, Malaika Watanabe, Ngah Zasmy Unyah, Sharif Alhassan Abdullahi, Ashraf Ahmad Issa Alapid, Norshariza Nordin, Rusliza Basir, Roslaini Abd Majid
      First page: 576
      Abstract: The major route for Toxoplasma gondii (T. gondii) infection is through the ingestion of foods contaminated with oocyst from cat faeces. The microscopic detection of T. gondii oocysts in cat faeces is challenging, which contributes to the failure of detecting or differentiating it from other related coccidian parasites. This study aims to detect T. gondii oocysts in cat faeces using two multicopy-target PCR assays and to evaluate their genetic diversity. Cat faecal (200) samples were collected from pet cats (PCs; 100) and free-roaming cats (FRCs; 100) within Klang Valley, Malaysia, and screened for coccidian oocysts by microscopy using Sheather’s sucrose floatation. PCR assays were performed on each faecal sample, targeting a B1 gene and a repetitive element (REP) gene to confirm T. gondii oocysts. Additionally, the PCR amplicons from the REP gene were sequenced to further confirm T. gondii-positive samples for phylogenetic analysis. Microscopy detected 7/200 (3.5%) T. gondii-like oocysts, while both the B1 gene and the REP gene detected 17/200 (8.5%) samples positive for T. gondii. All samples that were microscopically positive for T. gondii-like oocysts were also shown to be positive by both B1 and REP genes. The BLAST results sequenced for 16/200 (8.0%) PCR-positive T. gondii samples revealed homology and genetic heterogeneity with T. gondii strains in the GenBank, except for only one positive sample that did not show a result. There was almost perfect agreement (k = 0.145) between the two PCR assays targeting the B1 gene and the REP gene. This is the first report on microscopic, molecular detection and genetic diversity of T. gondii from cat faecal samples in Malaysia. In addition, the sensitivities of either the B1 gene or REP gene multicopy-target PCR assays are suitable for the accurate detection of T. gondii from cat faeces.
      Citation: Pathogens
      PubDate: 2020-07-16
      DOI: 10.3390/pathogens9070576
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 577: Polycystic Kidney Disease Ryanodine Receptor
           Domain (PKDRR) Proteins in Oomycetes

    • Authors: Limian Zheng, Barbara Doyle Prestwich, Patrick T. Harrison, John J. Mackrill
      First page: 577
      Abstract: In eukaryotes, two sources of Ca2+ are accessed to allow rapid changes in the cytosolic levels of this second messenger: the extracellular medium and intracellular Ca2+ stores, such as the endoplasmic reticulum. One class of channel that permits Ca2+ entry is the transient receptor potential (TRP) superfamily, including the polycystic kidney disease (PKD) proteins, or polycystins. Channels that release Ca2+ from intracellular stores include the inositol 1,4,5-trisphosphate/ryanodine receptor (ITPR/RyR) superfamily. Here, we characterise a family of proteins that are only encoded by oomycete genomes, that we have named PKDRR, since they share domains with both PKD and RyR channels. We provide evidence that these proteins belong to the TRP superfamily and are distinct from the ITPR/RyR superfamily in terms of their evolutionary relationships, protein domain architectures and predicted ion channel structures. We also demonstrate that a hypothetical PKDRR protein from Phytophthora infestans is produced by this organism, is located in the cell-surface membrane and forms multimeric protein complexes. Efforts to functionally characterise this protein in a heterologous expression system were unsuccessful but support a cell-surface localisation. These PKDRR proteins represent potential targets for the development of new “fungicides”, since they are of a distinctive structure that is only found in oomycetes and not in any other cellular organisms.
      Citation: Pathogens
      PubDate: 2020-07-16
      DOI: 10.3390/pathogens9070577
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 578: Prevalence of Infectious Spleen and Kidney
           Necrosis Virus (ISKNV), Nervous Necrosis Virus (NNV) and Ectoparasites in
           Juvenile Epinephelus spp. Farmed in Aceh, Indonesia

    • Authors: Bakhtiar Sah Putra, Paul M. Hick, Evelyn Hall, Richard J. Whittington, Razi Khairul, Evarianti, Nurbariah, Joy A. Becker
      First page: 578
      Abstract: A cross-sectional survey was used to estimate the prevalence of infections with the Infectious spleen and kidney necrosis virus (ISKNV, Megalocytivirus), nervous necrosis virus (NNV, Betanodavirus), and infestations with ectoparasites during the rainy season in juvenile grouper (Epinephelus spp.) farmed in Aceh, Indonesia. The survey was intended to detect aquatic pathogens present at 10% prevalence with 95% confidence, assuming 100% sensitivity and specificity using a sample size of 30 for each diagnostic test. Eight populations of grouper from seven farms were sampled. Additional targeted sampling was conducted for populations experiencing high mortality. Infection with NNV was detected at all farms with seven of the eight populations being positive. The apparent prevalence for NNV ranged from 0% (95% CI: 0–12) to 73% (95% CI: 54–88). All of the fish tested from the targeted samples (Populations 9 and 10) were positive for NNV and all had vacuolation of the brain and retina consistent with viral nervous necrosis (VNN). Coinfections with ISKNV were detected in five populations, with the highest apparent prevalence being 13% (95% CI: 4–31%). Trichodina sp., Cryptocaryon irritans and Gyrodactylus sp. were detected at three farms, with 66% to 100% of fish being infested. Hybrid grouper sourced from a hatchery were 5.4 and 24.9 times more likely to have a NNV infection and a higher parasite load compared to orange-spotted grouper collected from the wild (p < 0.001). This study found that VNN remains a high-impact disease in grouper nurseries in Aceh, Indonesia.
      Citation: Pathogens
      PubDate: 2020-07-16
      DOI: 10.3390/pathogens9070578
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 579: Pichinde Virus Infection of Outbred Hartley
           Guinea Pigs as a Surrogate Animal Model for Human Lassa Fever:
           Histopathological and Immunohistochemical Analyses

    • Authors: Wun-Ju Shieh, Shuiyun Lan, Sherif R. Zaki, Hinh Ly, Yuying Liang
      First page: 579
      Abstract: Lassa virus (LASV) is a mammarenavirus (arenavirus) that causes zoonotic infection in humans that can lead to fatal hemorrhagic Lassa fever (LF) disease. Currently, there are no FDA-approved vaccines or therapeutics against LASV. Development of treatments against LF and other related arenavirus-induced hemorrhagic fevers (AHFs) requires relevant animal models that can recapitulate clinical and pathological features of AHF diseases in humans. Laboratory mice are generally resistant to LASV infection, and non-human primates, while being a good animal model for LF, are limited by their high cost. Here, we describe a small, affordable, and convenient animal model that is based on outbred Hartley guinea pigs infected with Pichinde virus (PICV), a mammarenavirus that is non-pathogenic in humans, for use as a surrogate model of human LF. We conducted a detailed analysis of tissue histopathology and immunohistochemical analysis of different organs of outbred Hartley guinea pigs infected with different PICV strains that show differential disease phenotypes and pathologies. Comparing to infection with the avirulent PICV strain (P2 or rP2), animals infected with the virulent strain (P18 or rP18) show extensive pathological changes in different organs that sustain high levels of virus replication. The similarity of tissue pathology and viral antigen distribution between the virulent PICV–guinea pig model and lethal human LASV infection supports a role of this small animal model as a surrogate model of studying human LF in order to understand its pathogenesis and for evaluating potential preventative and therapeutic options against AHFs.
      Citation: Pathogens
      PubDate: 2020-07-16
      DOI: 10.3390/pathogens9070579
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 580: Genetic Diversity Among SARS-CoV2 Strains in
           South America may Impact Performance of Molecular Detection

    • Authors: Juan David Ramírez, Marina Muñoz, Carolina Hernández, Carolina Flórez, Sergio Gomez, Angelica Rico, Lisseth Pardo, Esther C. Barros, Alberto E. Paniz-Mondolfi
      First page: 580
      Abstract: Since its emergence in Wuhan (China) on December 2019, the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) has rapidly spread worldwide. After its arrival in South America in February 2020, the virus has expanded throughout the region, infecting over 900,000 individuals with approximately 41,000 reported deaths to date. In response to the rapidly growing number of cases, a number of different primer-probe sets have been developed. However, despite being highly specific, most of these primer-probe sets are known to exhibit variable sensitivity. Currently, there are more than 300 SARS-CoV2 whole genome sequences deposited in databases from Brazil, Chile, Ecuador, Colombia, Uruguay, Peru, and Argentina. To test how regional viral diversity may impact oligo binding sites and affect test performance, we reviewed all available primer-probe sets targeting the E, N, and RdRp genes against available South American SARS-CoV-2 genomes checking for nucleotide variations in annealing sites. Results from this in silico analysis showed no nucleotide variations on the E-gene target region, in contrast to the N and RdRp genes which showed massive nucleotide variations within oligo binding sites. In lines with previous data, our results suggest that the E-gene stands as the most conserved and reliable target when considering single-gene target testing for molecular diagnosis of SARS-CoV-2 in South America.
      Citation: Pathogens
      PubDate: 2020-07-17
      DOI: 10.3390/pathogens9070580
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 581: Contributions of Mass Spectrometry-Based
           Proteomics to Understanding Salmonella-Host Interactions

    • Authors: Buyu Zhang, Bohao Liu, Yinglin Zhou, Xinxiang Zhang, Qinghua Zou, Xiaoyun Liu
      First page: 581
      Abstract: As a model pathogen, Salmonella invades both phagocytic and non-phagocytic host cells and adopts an intracellular lifestyle in a membrane-bound compartment during infection. Therefore, a systemic overview of Salmonella adaptations to distinct host cells together with host remodeling will assist us in charting the landscape of host-pathogen interactions. Central to the Salmonella-host interplay are bacterial virulence factors (effectors) that are injected into host cells by type III secretion systems (T3SSs). Despite great progress, functional studies of bacterial effectors have experienced daunting challenges as well. In the last decade, mass spectrometry-based proteomics has evolved into a powerful technological platform that can quantitatively measure thousands of proteins in terms of their expression as well as post-translational modifications. Here, we will review the applications of high-throughput proteomic technologies in understanding the dynamic reprogramming of both Salmonella and host proteomes during the course of infection. Furthermore, we will summarize the progress in utilizing affinity purification-mass spectrometry to screen for host substrates of Salmonella T3SS effectors. Finally, we will critically discuss some limitations/challenges with current proteomic platforms in the context of host-pathogen interactions and highlight some emerging technologies that may offer the promise of tackling these problems.
      Citation: Pathogens
      PubDate: 2020-07-17
      DOI: 10.3390/pathogens9070581
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 582: Molecular Characterization of African Swine
           Fever Virus Isolates in Estonia in 2014–2019

    • Authors: Annika Vilem, Imbi Nurmoja, Tarmo Niine, Taavi Riit, Raquel Nieto, Arvo Viltrop, Carmina Gallardo
      First page: 582
      Abstract: After the extensive spread of the African swine fever virus (ASFV) genotype II in Eastern Europe, the first case of African swine fever (ASF) in Estonia was diagnosed in September 2014. By the end of 2019, 3971 ASFV-positive wild boars were found, and 27 domestic pig outbreaks were reported. A selection of ASFV isolates from wild boar and domestic pigs (during the period of September 2014–2019) was molecularly characterized using standardized genotyping procedures. One of the proven markers to characterize this virus is the central variable region (CVR) within the B602L gene. In summer 2015, a new ASFV genotype II CVR variant 2 (GII-CVR2) was confirmed in Estonia. The results suggest that the GII-CVR2 variant was only confirmed in wild boar from a limited area in southern Estonia in 2015 and 2016. In addition to GII-CVR2, a single nucleotide polymorphism (SNP) that resulted in amino acid change was identified within the genotype II CVR variant 1 (GII-CVR1). The GII-CVR1/SNP1 strain was isolated in Estonia in November 2016. Additional GII-CVR1/SNP1 cases were confirmed in two neighbouring counties, as well as in one outbreak farm in June 2017. Based on the available data, no GII-CVR2 and GII-CVR1/SNP1 have been reported by other affected European countries. The spread of variant strains in Estonia has been limited over time, and restricted to a relatively small area.
      Citation: Pathogens
      PubDate: 2020-07-17
      DOI: 10.3390/pathogens9070582
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 583: Bovine Papillomatosis Hiding a Zoonotic
           Infection: Epitheliotropic Viruses in Bovine Skin Lesions

    • Authors: Gallina, Savini, Canziani, Frasnelli, Lavazza, Scagliarini, Lelli
      First page: 583
      Abstract: We describe two cases of skin co-infections with epitheliotropic viruses, detected in two cattle during lumpy skin disease (LSD) surveillance in northern Italy. A diagnostic protocol including different molecular methods as well as negative staining electron microscopy was applied to detect the most common viral agents belonging to the family Papillomaviridae, Poxviridae and Herpesviridae which cause skin diseases in cattle. Two specimens were collected from cases clinically diagnosed as papillomatosis and pseudo-LSD. Both skin lesions were shown to harbor more than one viral species. This case report shows, for the first time, co-infection of zoonotic parapoxvirus with bovine papillomavirus and herpesvirus in skin lesions of cattle. In particular, the simultaneous presence of virions morphologically referable to parapoxvirus and papillomavirus confirms that the replication of both viruses in the same lesion can happen and the so-called papillomatosis can bear zoonotic viruses.
      Citation: Pathogens
      PubDate: 2020-07-17
      DOI: 10.3390/pathogens9070583
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 584: Polymorphism and Selection Pressure of
           SARS-CoV-2 Vaccine and Diagnostic Antigens: Implications for Immune
           Evasion and Serologic Diagnostic Performance

    • Authors: Eric Dumonteil, Claudia Herrera
      First page: 584
      Abstract: The ongoing SARS-CoV-2 pandemic has triggered multiple efforts for serological tests and vaccine development. Most of these tests and vaccines are based on the Spike glycoprotein (S) or the Nucleocapsid (N) viral protein. Conservation of these antigens among viral strains is critical to ensure optimum diagnostic test performance and broad protective efficacy, respectively. We assessed N and S antigen diversity from 17,853 SARS-CoV-2 genome sequences and evaluated selection pressure. Up to 6–7 incipient phylogenetic clades were identified for both antigens, confirming early variants of the S antigen and identifying new ones. Significant diversifying selection was detected at multiple sites for both antigens. Some sequence variants have already spread in multiple regions, in spite of their low frequency. In conclusion, the N and S antigens of SARS-CoV-2 are well-conserved antigens, but new clades are emerging and may need to be included in future diagnostic and vaccine formulations.
      Citation: Pathogens
      PubDate: 2020-07-17
      DOI: 10.3390/pathogens9070584
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 585: Scientometrics Approach to Research in Ovine
           Mastitis from 1970 to 2019 (with a Complete List of Relevant Literature

    • Authors: Daphne T. Lianou, George C. Fthenakis
      First page: 585
      Abstract: The present study is a scientometrics evaluation of refereed publications on bacterial mastitis in sheep; the objectives were the evaluation of the relevant papers and the presentation of quantitative characteristics regarding their scientific content and bibliometric details. The Web of Science platform was used with search terms: [mastitis OR *mammary infection*] AND [sheep OR ewe* OR ovine] for papers from 1970 tο 2019; only ‘articles’, ‘reviews’, ‘proceedings papers’, or ‘data papers’ were evaluated, whilst documents related solely to contagious agalactia, mammary aspects of lentiviral infections, or infections of the teats and the udder skin were excluded. Finally, 580 papers were considered in detail. The number of published papers increased from 8 during the 1970s to 273 during the 2010s. These papers originated from 43 countries (most from Greece or Spain, n = 87 from each) and 240 institutions (145 universities and 95 other establishments), of which 35 produced ≥ 5 papers each. Most papers present original studies (n = 539) with a few reviews (n = 41). The original papers refer to dairy (n = 428), meat (n = 113), or wool (n = 1) production systems and present field (n = 329), laboratory (n = 163), or experimental (n = 67) work; the papers report aetiology (n = 146), risk factors (n = 100), pathogenesis (n = 92), diagnosis (n = 88), effects (n = 66), treatment (n = 50), control (n = 36), or descriptive epidemiology (n = 32) of the disease. Papers related to dairy production present more field and fewer experimental work than papers related to meat production; also, in papers describing work performed in dairy sheep, studies about aetiology, risk factors, and diagnosis of the disease predominate, whilst in papers performed in meat sheep, studies about aetiology, pathogenesis, and effects/diagnosis are reported more often. The papers were published in 175 scientific journals (most in Small Ruminant Research, n = 90, or Journal of Dairy Science, n = 54). On average, the papers received 16.8 total citations and 1.6 yearly citations (h-index = 47). Most papers were published in Scimago classification Q1 (n = 240) or Q2 (n = 230) journals and received 23.4 or 15.4 total citations, respectively. Reviews received more citations than original papers; among the latter, papers with work referring to dairy production received more yearly citations than papers referring to meat production; no differences in citations were seen according to type of work or mastitis aspect covered. Most citations were received by papers from France. Papers published in Journal of Dairy Science or Small Ruminant Research received the most citations. In total, there were 1558 individual authors of the papers, with 24 authors having co-authored > 10 papers each (max: 73 papers); on average, there were 5.2 co-authors per paper (min–max: 1–25). Average number of co-authors progressively increased from 2.1 in the 1970s to 6.3 in the 2010s, with original papers having a higher number of co-authors than reviews: 5.3 and 3.7, respectively. Papers from France had highers number of co-authors (7.9). The findings of this first ever scientometrics study into ovine mastitis indicate that the disease has not been studied as other sheep diseases and that future studies in it should be directed to its control.
      Citation: Pathogens
      PubDate: 2020-07-17
      DOI: 10.3390/pathogens9070585
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 586: Stenamoeba dejonckheerei sp. nov., a
           Free-Living Amoeba Isolated from a Thermal Spring

    • Authors: Manuel Alejandro Borquez-Román, Luis Fernando Lares-Jiménez, Libia Zulema Rodriguez-Anaya, Jose Reyes Gonzalez-Galaviz, Paul A. Fuerst, José Cuauhtémoc Ibarra-Gámez, Ramón Casillas-Hernández, Fernando Lares-Villa
      First page: 586
      Abstract: Two amoeboid organisms were obtained from water samples taken from a thermal spring, "Agua Caliente", in Northwestern Mexico. The isolates were obtained when samples were cultivated at 37 °C on non-nutrient agar coated with Escherichia coli. The initial identification of the isolates was performed morphologically using light microscopy. The samples were found to have trophozoite morphology consistent with members of the genus Stenamoeba, a genus derived in 2007 from within the abolished polyphyletic genus Platyamoeba. Further analysis was performed by sequencing PCR products obtained using universal eukaryotic primers for the small subunit ribosomal ribonucleic acid (SSU rRNA) gene. Sequencing primers were designed to allow the comparison of the 18S rRNA gene sequences of the new isolates with previous sequences reported for Stenamoeba. Phylogenetic relationships among sequences from Stenamoeba were determined using Maximum Likelihood analysis. The results showed the two "Agua Caliente" sequences to be closely related, while clearly separating them from those of other Stenamoeba taxa. The degrees of sequence differentiation from other taxa were considered sufficient to allow us to propose that the Mexican isolates represent a new species.
      Citation: Pathogens
      PubDate: 2020-07-17
      DOI: 10.3390/pathogens9070586
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 587: Modulation of Leptin and Leptin Receptor
           Expression in Mice Acutely Infected with Neospora caninum

    • Authors: Luzia Teixeira, Alexandra Correia, Bárbara M. Oliveira, Ana Pinto, Paula G. Ferreira, Manuel Vilanova
      First page: 587
      Abstract: Neospora caninum is an apicomplexan parasite that in cattle assumes particular importance, as it is responsible for abortions reported worldwide. Leptin is an adipokine mainly secreted by adipocytes, which beside its role in maintaining metabolic homeostasis also has important effects in both innate and adaptive immunity. In previous work, we showed that mice chronically infected with N. caninum had elevated serum leptin levels. Here, we sought to assess whether acute infection with N. caninum infection influenced the production of this adipokine as well as leptin receptor mRNA levels. Our results show that acute infection with N. caninum led to decreased leptin serum levels and mRNA expression in adipose tissue. A decrease in leptin receptor transcript variant 1 mRNA (long isoform) and leptin receptor transcript variant 3 mRNA (one of the short isoforms) expression was also observed. An increase in the number of cells staining positive for leptin in the liver of infected mice was observed, although this increase was less marked in Interleukin (IL)-12/IL-23 p40-deficient mice. Overall, our results show that N. caninum infection also influences leptin production during acute infection.
      Citation: Pathogens
      PubDate: 2020-07-17
      DOI: 10.3390/pathogens9070587
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 588: Identification of Antimicrobial
           Resistance-Associated Genes through Whole Genome Sequencing of Mycoplasma
           bovis Isolates with Different Antimicrobial Resistances

    • Authors: Lisa Ledger, Jason Eidt, Hugh Y. Cai
      First page: 588
      Abstract: Antimicrobial resistance (AMR) in Mycoplasma bovis has been previously associated with topoisomerase and ribosomal gene mutations rather than specific resistance-conferring genes. Using whole genome sequencing (WGS) to identify potential new AMR mechanisms for M. bovis, it was found that a 2019 clinical isolate with high MIC (2019-043682) for fluoroquinolones, macrolides, lincosamides, pleuromutilins and tetracyclines had a new core genome multilocus sequencing (cgMLST) type (ST10-like) and 91% sequence similarity to the published genome of M. bovis PG45. Closely related to PG45, a 1982 isolate (1982-M6152) shared the same cgMLST type (ST17), 97.2% sequence similarity and low MIC results. Known and potential AMR- associated genetic events were identified through multiple sequence alignment of the three genomes. Isolate 2019-043682 had 507 genes with non-synonymous mutations (NSMs) and 67 genes disrupted. Isolate 1982-M6152 had 81 NSMs and 20 disruptions. Using functional roles and known mechanisms of antimicrobials, a 55 gene subset was assessed for AMR potential. Seventeen were previously identified from other bacteria as sites of AMR mutation, 38 shared similar functions to them, and 11 contained gene-disrupting mutations. This study indicated that M. bovis may obtain high AMR characteristics by mutating or disrupting other functional genes, in addition to topoisomerases and ribosomal genes.
      Citation: Pathogens
      PubDate: 2020-07-19
      DOI: 10.3390/pathogens9070588
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 589: West Nile Virus: An Update on Pathobiology,
           Epidemiology, Diagnostics, Control and “One Health” Implications

    • Authors: Gervais Habarugira, Willy W. Suen, Jody Hobson-Peters, Roy A. Hall, Helle Bielefeldt-Ohmann
      First page: 589
      Abstract: West Nile virus (WNV) is an important zoonotic flavivirus responsible for mild fever to severe, lethal neuroinvasive disease in humans, horses, birds, and other wildlife species. Since its discovery, WNV has caused multiple human and animal disease outbreaks in all continents, except Antarctica. Infections are associated with economic losses, mainly due to the cost of treatment of infected patients, control programmes, and loss of animals and animal products. The pathogenesis of WNV has been extensively investigated in natural hosts as well as in several animal models, including rodents, lagomorphs, birds, and reptiles. However, most of the proposed pathogenesis hypotheses remain contentious, and much remains to be elucidated. At the same time, the unavailability of specific antiviral treatment or effective and safe vaccines contribute to the perpetuation of the disease and regular occurrence of outbreaks in both endemic and non-endemic areas. Moreover, globalisation and climate change are also important drivers of the emergence and re-emergence of the virus and disease. Here, we give an update of the pathobiology, epidemiology, diagnostics, control, and “One Health” implications of WNV infection and disease.
      Citation: Pathogens
      PubDate: 2020-07-19
      DOI: 10.3390/pathogens9070589
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 590: Understanding HCMV Latency Using Unbiased
           Proteomic Analyses

    • Authors: Emma Poole, John Sinclair
      First page: 590
      Abstract: Human cytomegalovirus (HCMV) establishes either a latent (non-productive) or lytic (productive) infection depending upon cell type, cytokine milieu and the differentiation status of the infected cell. Undifferentiated cells, such as precursor cells of the myeloid lineage, support a latent infection whereas terminally differentiated cells, such as monocytes or dendritic cells are an environment conducive to reactivation and support a lytic infection. The mechanisms which regulate HCMV in either a latent or lytic infection have been the focus of intense investigation with a view to developing novel treatments for HCMV-associated disease which can have a heavy clinical burden after reactivation or primary infection in, especially, the immune compromised. To this end, a number of studies have been carried out in an unbiased manner to address global changes occurring within the latently infected cell to address the molecular changes associated with HCMV latency. In this review, we will concentrate on the proteomic analyses which have been carried out in undifferentiated myeloid cells which either stably express specific viral latency associated genes in isolation or on cells which have been latently infected with virus.
      Citation: Pathogens
      PubDate: 2020-07-20
      DOI: 10.3390/pathogens9070590
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 591: Concurrent Infection of Skunk Adenovirus-1,
           Listeria monocytogenes, and a Regionally Specific Clade of Canine
           Distemper Virus in One Gray Fox (Urocyon cinereoargenteus) and Concurrent
           Listeriosis and Canine Distemper in a Second Gray Fox

    • Authors: David B. Needle, Jacqueline L. Marr, Cooper J. Park, Cheryl P. Andam, Annabel G. Wise, Roger K. Maes, Rebecca P. Wilkes, Eman A. Anis, Inga F. Sidor, Dalen Agnew, Julie C. Ellis, Patrick Tate, Abigail Mathewson, Christopher Benton, Robert Gibson
      First page: 591
      Abstract: One free-ranging Gray fox (Urocyon cinereoargenteus) underwent autopsy following neurologic disease, with findings including morbilliviral inclusions and associated lesions in numerous tissues, adenoviral intranuclear inclusions in bronchial epithelial cells, and septic pleuropneumonia, hepatitis, splenitis, and meningoencephalitis. Molecular diagnostics on fresh lung identified a strain within a distinct clade of canine distemper that is currently unique to wildlife in New England, as well as the emerging multi-host viral pathogen skunk adenovirus-1. Bacterial culture of fresh liver resulted in a pure growth of Listeria monocytogenes, with whole genome sequencing indicating that the isolate had a vast array of antimicrobial resistance and virulence-associated genes. One year later, a second fox was euthanized for inappropriate behavior in a residential area, and diagnostic workup revealed canine distemper and septic L. monocytogenes, with the former closely related to the distemper virus found in the previous fox and the latter divergent from the L. monocytogenes from the previous fox.
      Citation: Pathogens
      PubDate: 2020-07-21
      DOI: 10.3390/pathogens9070591
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 592: Mycobacterium bovis Population Structure in
           Cattle and Local Badgers: Co-Localisation and Variation by Farm Type

    • Authors: Georgina Milne, Adrian Allen, Jordon Graham, Raymond Kirke, Carl McCormick, Eleanor Presho, Robin Skuce, Andrew W. Byrne
      First page: 592
      Abstract: Bovine tuberculosis surveillance in Northern Ireland includes Multiple-Locus Variable number tandem repeat Analysis (MLVA) to determine the Mycobacterium bovis genetic type present in both cattle and the predominant wildlife host, the European badger (Meles meles). These data are useful for investigating clusters of infection and understanding the scale at which interspecific transmission may occur. We utilised a comprehensive dataset of routinely sampled isolates from infected cattle and from badgers killed in road-traffic accidents to investigate the spatial co-location of MLVA types in, and between, the badger and cattle populations. Furthermore, we investigated the hypothesis that the type of farming enterprise might explain some variation in this relationship. MLVA types were spatially co-localised in cattle and road-traffic accident (RTA) badger hosts, indicative of a shared epidemic. Dairy herds were more likely to have at least one MLVA type in common with nearby RTA badgers, compared to non-dairy herd types. Marginally more MLVA spatial clustering was observed in non-dairy herds, which may be a consequence of relatively more between-herd movements. For the cattle population, local transmission mechanisms such as infection from contiguous herds, infectious wildlife and short-range between-herd cattle movements appear primarily to drive the epidemic: there appears to be a more limited role for long-range movements. Animal management practices are likely to be the driving force behind this observation, as beef rearing is associated with elevated numbers of animal movements compared to dairy herds.
      Citation: Pathogens
      PubDate: 2020-07-21
      DOI: 10.3390/pathogens9070592
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 593: Monitoring Mycoplasma bovis Diversity and
           Antimicrobial Susceptibility in Calf Feedlots Undergoing a Respiratory
           Disease Outbreak

    • Authors: Claire A.M. Becker, Chloé Ambroset, Anthéa Huleux, Angélique Vialatte, Adélie Colin, Agnès Tricot, Marie-Anne Arcangioli, Florence Tardy
      First page: 593
      Abstract: Bovine respiratory diseases (BRD) are widespread in veal calf feedlots. Several pathogens are implicated, both viruses and bacteria, one of which, Mycoplasma bovis, is under-researched. This worldwide-distributed bacterium has been shown to be highly resistant in vitro to the main antimicrobials used to treat BRD. Our objective was to monitor the relative prevalence of M. bovis during BRD episodes, its diversity, and its resistance phenotype in relation to antimicrobial use. For this purpose, a two-year longitudinal follow-up of 25 feedlots was organized and 537 nasal swabs were collected on 358 veal calves at their arrival in the lot, at the BRD peak and 4 weeks after collective antimicrobial treatments. The presence of M. bovis was assessed by real-time PCR and culture. The clones isolated were then subtyped (polC subtyping and PFGE analysis), and their susceptibility to five antimicrobials was determined. The course of the disease and the antimicrobials used had no influence on the genetic diversity of the M. bovis strains: The subtype distribution was the same throughout the BRD episode and similar to that already described in France, with a major narrowly-variable subtype circulating, st2. The same conclusion holds for antimicrobial resistance (AMR) phenotypes: All the clones were already multiresistant to the main antimicrobials used (except for fluoroquinolones) prior to any treatments. By contrast, changes of AMR phenotypes could be suspected for Pasteurellaceae in two cases in relation to the treatments registered.
      Citation: Pathogens
      PubDate: 2020-07-21
      DOI: 10.3390/pathogens9070593
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 594: The Epstein-Barr Virus-Encoded EBNA1 Protein
           Activates the Bone Morphogenic Protein (BMP) Signalling Pathway to Promote
           Carcinoma Cell Migration

    • Authors: Hannah E. Bridgewater, Kathryn L. Date, John D. O’Neil, Chunfang Hu, John R. Arrand, Christopher W. Dawson, Lawrence S. Young
      First page: 594
      Abstract: The Epstein-Barr virus (EBV)-encoded nuclear antigen 1 (EBNA1) protein is expressed in all virus-associated malignancies, where it performs an essential role in the maintenance, replication and transcription of the EBV genome. In recent years, it has become apparent that EBNA1 can also influence cellular gene transcription. Here, we demonstrate that EBNA1 is able to stimulate the expression of the Transforming growth factor-beta (TGFβ) superfamily member, bone morphogenic protein 2 (BMP2), with consequential activation of the BMP signalling pathway in carcinoma cell lines. We show that BMP pathway activation is associated with an increase in the migratory capacity of carcinoma cells, an effect that can be ablated by the BMP antagonist, Noggin. Gene expression profiling of authentic EBV-positive nasopharyngeal carcinoma (NPC) tumours revealed the consistent presence of BMP ligands, established BMP pathway effectors and putative target genes, constituting a prominent BMP “signature” in this virus-associated cancer. Our findings show that EBNA1 is the major viral-encoded protein responsible for activating the BMP signalling pathway in carcinoma cells and supports a role for this pathway in promoting cell migration and possibly, metastatic spread.
      Citation: Pathogens
      PubDate: 2020-07-21
      DOI: 10.3390/pathogens9070594
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 595: First Report on the Occurrence and Subtypes
           of Blastocystis in Pigs in Poland Using Sequence-Tagged-Site PCR and
           Barcode Region Sequencing

    • Authors: Monika Rudzińska, Beata Kowalewska, Beata Szostakowska, Maciej Grzybek, Katarzyna Sikorska, Agnieszka Świątalska
      First page: 595
      Abstract: Blastocystis is an enteric microorganism commonly found in humans and animals worldwide. Its pathogenic role in humans and transmission patterns has not been fully explained. However, nine subtypes (ST1–8, ST12) are considered as potentially zoonotic. Studies from various regions of the world show that pigs are mainly infected with ST5. Although pigs are important farmed animals in Poland, the question of Blastocystis infection in these animals has not yet been investigated. Herein, 149 pig stool samples from 10 Polish pig farms were analyzed using sequence-tagged-site PCR and barcode region sequencing. The percentage of samples in which Blastocystis was identified using each method separately was similar: 38.25% and 37.58%, respectively. However, the percentage of positive results obtained by combining both methods was 46.97%, which means that, depending on the method used, the number of undetected samples varied between 8.72% and 9.39%. This shows the methodological limitations of up-to-date molecular approaches commonly used in Blastocystis research. A moderate infection rate (44.4–50%) observed in different pig age groups with a vital predominance of ST5 (94.28%) in every age group shows that pigs are a likely natural host of ST5. A small percentage of mixed infections, namely ST5/ST1 (5.26%), ST5/ST3 (1.75%), and ST3/ST1 (1.75%), was observed only in animals of older age, suggesting that ST3 and ST1 can be acquired by pigs during contact with humans. This study provides the first data on the prevalence and Blastocystis subtypes (STs) distribution in pigs in Poland. The results also highlight the need for the development of new methods capable of detecting highly genetically diverse Blastocystis isolates and mixed infections.
      Citation: Pathogens
      PubDate: 2020-07-21
      DOI: 10.3390/pathogens9070595
      Issue No: Vol. 9, No. 7 (2020)
  • Pathogens, Vol. 9, Pages 596: Molecular Characterization of Clistobothrium
           sp. Viable Plerocercoids in Fresh Longfin Inshore Squid (Doryteuthis
           pealeii) and Implications for Cephalopod Inspection

    • Authors: Guardone, Giusti, Bilska-Zajac, Malandra, Różycki, Armani
      First page: 596
      Abstract: Cephalopods, an appreciated seafood product, are common hosts of marine cestodes. The aim of this work is to report visible alive plerocercoids in longfin inshore squid (Doryteuthis pealeii), a cephalopod species commercialized as fresh and whole in Italy. Seventy D. pealeii from the Northwest Atlantic (FAO area 21) were collected and visually inspected. In total, 18 plerocercoid larvae were found in the viscera of 10 host specimens (P: 14.3% 95% CI 7.1–24.7; MI: 1.8, MA: 0.26; range 1–4) and molecularly analyzed targeting the variable D2 region of the large subunit (LSU) rRNA gene and the cytochrome c oxidase subunit I (COI) gene. The molecular characterization allowed to identify all the plerocercoids as Clistobothrium sp., a cestode of the Phyllobothriidae family with Lamnidae sharks as definitive hosts, and cephalopods as second intermediate hosts. These findings represent the first molecular record of Clistobothrium sp. in D. pealeii, thus contributing to elucidate its poorly known life cycle. Even if not affecting consumer’s health, these visible parasites may represent a reason for disgust for consumers. Therefore, the results suggest that Food Business Operators should also check for the presence of these visible parasites during inspection and underline the importance of a correct consumers’ education.
      Citation: Pathogens
      PubDate: 2020-07-21
      DOI: 10.3390/pathogens9070596
      Issue No: Vol. 9, No. 7 (2020)
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