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Showing 1 - 200 of 3175 Journals sorted alphabetically
A Practical Logic of Cognitive Systems     Full-text available via subscription   (Followers: 8)
AASRI Procedia     Open Access   (Followers: 14)
Academic Pediatrics     Hybrid Journal   (Followers: 28, SJR: 1.402, h-index: 51)
Academic Radiology     Hybrid Journal   (Followers: 22, SJR: 1.008, h-index: 75)
Accident Analysis & Prevention     Partially Free   (Followers: 90, SJR: 1.109, h-index: 94)
Accounting Forum     Hybrid Journal   (Followers: 25, SJR: 0.612, h-index: 27)
Accounting, Organizations and Society     Hybrid Journal   (Followers: 33, SJR: 2.515, h-index: 90)
Achievements in the Life Sciences     Open Access   (Followers: 5)
Acta Anaesthesiologica Taiwanica     Open Access   (Followers: 6, SJR: 0.338, h-index: 19)
Acta Astronautica     Hybrid Journal   (Followers: 376, SJR: 0.726, h-index: 43)
Acta Automatica Sinica     Full-text available via subscription   (Followers: 2)
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Acta de Investigación Psicológica     Open Access   (Followers: 3)
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Acta Histochemica     Hybrid Journal   (Followers: 3, SJR: 0.604, h-index: 38)
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Acta Oecologica     Hybrid Journal   (Followers: 10, SJR: 0.915, h-index: 53)
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Acta Pharmaceutica Sinica B     Open Access   (Followers: 1)
Acta Poética     Open Access   (Followers: 4)
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Ad Hoc Networks     Hybrid Journal   (Followers: 11, SJR: 0.967, h-index: 57)
Addictive Behaviors     Hybrid Journal   (Followers: 15, SJR: 1.514, h-index: 92)
Addictive Behaviors Reports     Open Access   (Followers: 7)
Additive Manufacturing     Hybrid Journal   (Followers: 9, SJR: 1.039, h-index: 5)
Additives for Polymers     Full-text available via subscription   (Followers: 22)
Advanced Cement Based Materials     Full-text available via subscription   (Followers: 3)
Advanced Drug Delivery Reviews     Hybrid Journal   (Followers: 128, SJR: 5.2, h-index: 222)
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Advances in Child Development and Behavior     Full-text available via subscription   (Followers: 10, SJR: 0.665, h-index: 29)
Advances in Chronic Kidney Disease     Full-text available via subscription   (Followers: 10, SJR: 1.268, h-index: 45)
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Advances in Engineering Software     Hybrid Journal   (Followers: 27, SJR: 0.486, h-index: 10)
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Advances in Experimental Social Psychology     Full-text available via subscription   (Followers: 42, SJR: 5.465, h-index: 64)
Advances in Exploration Geophysics     Full-text available via subscription   (Followers: 1)
Advances in Fluorine Science     Full-text available via subscription   (Followers: 7)
Advances in Food and Nutrition Research     Full-text available via subscription   (Followers: 54, SJR: 0.674, h-index: 38)
Advances in Fuel Cells     Full-text available via subscription   (Followers: 15)
Advances in Genetics     Full-text available via subscription   (Followers: 14, SJR: 2.558, h-index: 54)
Advances in Genome Biology     Full-text available via subscription   (Followers: 7)
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Advances in Heat Transfer     Full-text available via subscription   (Followers: 21, SJR: 0.906, h-index: 24)
Advances in Heterocyclic Chemistry     Full-text available via subscription   (Followers: 9, SJR: 0.497, h-index: 31)
Advances in Human Factors/Ergonomics     Full-text available via subscription   (Followers: 23)
Advances in Imaging and Electron Physics     Full-text available via subscription   (Followers: 1, SJR: 0.396, h-index: 27)
Advances in Immunology     Full-text available via subscription   (Followers: 36, SJR: 4.152, h-index: 85)
Advances in Inorganic Chemistry     Full-text available via subscription   (Followers: 8, SJR: 1.132, h-index: 42)
Advances in Insect Physiology     Full-text available via subscription   (Followers: 2, SJR: 1.274, h-index: 27)
Advances in Integrative Medicine     Hybrid Journal   (Followers: 6)
Advances in Intl. Accounting     Full-text available via subscription   (Followers: 3)
Advances in Life Course Research     Hybrid Journal   (Followers: 8, SJR: 0.764, h-index: 15)
Advances in Lipobiology     Full-text available via subscription   (Followers: 1)
Advances in Magnetic and Optical Resonance     Full-text available via subscription   (Followers: 9)
Advances in Marine Biology     Full-text available via subscription   (Followers: 14, SJR: 1.645, h-index: 45)
Advances in Mathematics     Full-text available via subscription   (Followers: 10, SJR: 3.261, h-index: 65)
Advances in Medical Sciences     Hybrid Journal   (Followers: 6, SJR: 0.489, h-index: 25)
Advances in Medicinal Chemistry     Full-text available via subscription   (Followers: 5)
Advances in Microbial Physiology     Full-text available via subscription   (Followers: 4, SJR: 1.44, h-index: 51)
Advances in Molecular and Cell Biology     Full-text available via subscription   (Followers: 21)
Advances in Molecular and Cellular Endocrinology     Full-text available via subscription   (Followers: 8)
Advances in Molecular Toxicology     Full-text available via subscription   (Followers: 7, SJR: 0.324, h-index: 8)
Advances in Nanoporous Materials     Full-text available via subscription   (Followers: 3)
Advances in Oncobiology     Full-text available via subscription   (Followers: 1)
Advances in Organ Biology     Full-text available via subscription   (Followers: 1)
Advances in Organometallic Chemistry     Full-text available via subscription   (Followers: 15, SJR: 2.885, h-index: 45)
Advances in Parallel Computing     Full-text available via subscription   (Followers: 6, SJR: 0.148, h-index: 11)
Advances in Parasitology     Full-text available via subscription   (Followers: 5, SJR: 2.37, h-index: 73)
Advances in Pediatrics     Full-text available via subscription   (Followers: 24, SJR: 0.4, h-index: 28)
Advances in Pharmaceutical Sciences     Full-text available via subscription   (Followers: 10)
Advances in Pharmacology     Full-text available via subscription   (Followers: 15, SJR: 1.718, h-index: 58)
Advances in Physical Organic Chemistry     Full-text available via subscription   (Followers: 8, SJR: 0.384, h-index: 26)
Advances in Phytomedicine     Full-text available via subscription  
Advances in Planar Lipid Bilayers and Liposomes     Full-text available via subscription   (Followers: 3, SJR: 0.248, h-index: 11)
Advances in Plant Biochemistry and Molecular Biology     Full-text available via subscription   (Followers: 7)
Advances in Plant Pathology     Full-text available via subscription   (Followers: 5)
Advances in Porous Media     Full-text available via subscription   (Followers: 5)
Advances in Protein Chemistry     Full-text available via subscription   (Followers: 17)
Advances in Protein Chemistry and Structural Biology     Full-text available via subscription   (Followers: 18, SJR: 1.5, h-index: 62)
Advances in Psychology     Full-text available via subscription   (Followers: 59)
Advances in Quantum Chemistry     Full-text available via subscription   (Followers: 6, SJR: 0.478, h-index: 32)
Advances in Radiation Oncology     Open Access  
Advances in Small Animal Medicine and Surgery     Hybrid Journal   (Followers: 3, SJR: 0.1, h-index: 2)
Advances in Space Biology and Medicine     Full-text available via subscription   (Followers: 5)
Advances in Space Research     Full-text available via subscription   (Followers: 375, SJR: 0.606, h-index: 65)
Advances in Structural Biology     Full-text available via subscription   (Followers: 5)
Advances in Surgery     Full-text available via subscription   (Followers: 9, SJR: 0.823, h-index: 27)
Advances in the Study of Behavior     Full-text available via subscription   (Followers: 29, SJR: 1.321, h-index: 56)
Advances in Veterinary Medicine     Full-text available via subscription   (Followers: 17)
Advances in Veterinary Science and Comparative Medicine     Full-text available via subscription   (Followers: 13)
Advances in Virus Research     Full-text available via subscription   (Followers: 5, SJR: 1.878, h-index: 68)
Advances in Water Resources     Hybrid Journal   (Followers: 46, SJR: 2.408, h-index: 94)
Aeolian Research     Hybrid Journal   (Followers: 6, SJR: 0.973, h-index: 22)
Aerospace Science and Technology     Hybrid Journal   (Followers: 333, SJR: 0.816, h-index: 49)
AEU - Intl. J. of Electronics and Communications     Hybrid Journal   (Followers: 8, SJR: 0.318, h-index: 36)
African J. of Emergency Medicine     Open Access   (Followers: 6, SJR: 0.344, h-index: 6)
Ageing Research Reviews     Hybrid Journal   (Followers: 9, SJR: 3.289, h-index: 78)
Aggression and Violent Behavior     Hybrid Journal   (Followers: 429, SJR: 1.385, h-index: 72)
Agri Gene     Hybrid Journal  
Agricultural and Forest Meteorology     Hybrid Journal   (Followers: 15, SJR: 2.18, h-index: 116)
Agricultural Systems     Hybrid Journal   (Followers: 31, SJR: 1.275, h-index: 74)
Agricultural Water Management     Hybrid Journal   (Followers: 43, SJR: 1.546, h-index: 79)
Agriculture and Agricultural Science Procedia     Open Access   (Followers: 1)
Agriculture and Natural Resources     Open Access   (Followers: 2)
Agriculture, Ecosystems & Environment     Hybrid Journal   (Followers: 56, SJR: 1.879, h-index: 120)
Ain Shams Engineering J.     Open Access   (Followers: 5, SJR: 0.434, h-index: 14)
Air Medical J.     Hybrid Journal   (Followers: 5, SJR: 0.234, h-index: 18)
AKCE Intl. J. of Graphs and Combinatorics     Open Access   (SJR: 0.285, h-index: 3)
Alcohol     Hybrid Journal   (Followers: 11, SJR: 0.922, h-index: 66)
Alcoholism and Drug Addiction     Open Access   (Followers: 9)
Alergologia Polska : Polish J. of Allergology     Full-text available via subscription   (Followers: 1)
Alexandria Engineering J.     Open Access   (Followers: 1, SJR: 0.436, h-index: 12)
Alexandria J. of Medicine     Open Access   (Followers: 1)
Algal Research     Partially Free   (Followers: 9, SJR: 2.05, h-index: 20)
Alkaloids: Chemical and Biological Perspectives     Full-text available via subscription   (Followers: 2)
Allergologia et Immunopathologia     Full-text available via subscription   (Followers: 1, SJR: 0.46, h-index: 29)
Allergology Intl.     Open Access   (Followers: 5, SJR: 0.776, h-index: 35)
Alpha Omegan     Full-text available via subscription   (SJR: 0.121, h-index: 9)
ALTER - European J. of Disability Research / Revue Européenne de Recherche sur le Handicap     Full-text available via subscription   (Followers: 9, SJR: 0.158, h-index: 9)
Alzheimer's & Dementia     Hybrid Journal   (Followers: 48, SJR: 4.289, h-index: 64)
Alzheimer's & Dementia: Diagnosis, Assessment & Disease Monitoring     Open Access   (Followers: 4)
Alzheimer's & Dementia: Translational Research & Clinical Interventions     Open Access   (Followers: 4)
Ambulatory Pediatrics     Hybrid Journal   (Followers: 6)
American Heart J.     Hybrid Journal   (Followers: 50, SJR: 3.157, h-index: 153)
American J. of Cardiology     Hybrid Journal   (Followers: 50, SJR: 2.063, h-index: 186)
American J. of Emergency Medicine     Hybrid Journal   (Followers: 42, SJR: 0.574, h-index: 65)
American J. of Geriatric Pharmacotherapy     Full-text available via subscription   (Followers: 10, SJR: 1.091, h-index: 45)
American J. of Geriatric Psychiatry     Hybrid Journal   (Followers: 14, SJR: 1.653, h-index: 93)
American J. of Human Genetics     Hybrid Journal   (Followers: 31, SJR: 8.769, h-index: 256)
American J. of Infection Control     Hybrid Journal   (Followers: 26, SJR: 1.259, h-index: 81)
American J. of Kidney Diseases     Hybrid Journal   (Followers: 32, SJR: 2.313, h-index: 172)
American J. of Medicine     Hybrid Journal   (Followers: 42, SJR: 2.023, h-index: 189)
American J. of Medicine Supplements     Full-text available via subscription   (Followers: 3)
American J. of Obstetrics and Gynecology     Hybrid Journal   (Followers: 189, SJR: 2.255, h-index: 171)
American J. of Ophthalmology     Hybrid Journal   (Followers: 62, SJR: 2.803, h-index: 148)
American J. of Ophthalmology Case Reports     Open Access   (Followers: 6)
American J. of Orthodontics and Dentofacial Orthopedics     Full-text available via subscription   (Followers: 6, SJR: 1.249, h-index: 88)
American J. of Otolaryngology     Hybrid Journal   (Followers: 25, SJR: 0.59, h-index: 45)
American J. of Pathology     Hybrid Journal   (Followers: 27, SJR: 2.653, h-index: 228)
American J. of Preventive Medicine     Hybrid Journal   (Followers: 27, SJR: 2.764, h-index: 154)
American J. of Surgery     Hybrid Journal   (Followers: 37, SJR: 1.286, h-index: 125)
American J. of the Medical Sciences     Hybrid Journal   (Followers: 12, SJR: 0.653, h-index: 70)
Ampersand : An Intl. J. of General and Applied Linguistics     Open Access   (Followers: 6)
Anaerobe     Hybrid Journal   (Followers: 4, SJR: 1.066, h-index: 51)
Anaesthesia & Intensive Care Medicine     Full-text available via subscription   (Followers: 61, SJR: 0.124, h-index: 9)
Anaesthesia Critical Care & Pain Medicine     Full-text available via subscription   (Followers: 14)
Anales de Cirugia Vascular     Full-text available via subscription  
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Anales de Pediatría (English Edition)     Full-text available via subscription  
Anales de Pediatría Continuada     Full-text available via subscription   (SJR: 0.104, h-index: 3)
Analytic Methods in Accident Research     Hybrid Journal   (Followers: 4, SJR: 2.577, h-index: 7)
Analytica Chimica Acta     Hybrid Journal   (Followers: 39, SJR: 1.548, h-index: 152)
Analytical Biochemistry     Hybrid Journal   (Followers: 165, SJR: 0.725, h-index: 154)
Analytical Chemistry Research     Open Access   (Followers: 10, SJR: 0.18, h-index: 2)
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Anesthésie & Réanimation     Full-text available via subscription   (Followers: 1)
Anesthesiology Clinics     Full-text available via subscription   (Followers: 22, SJR: 0.421, h-index: 40)
Angiología     Full-text available via subscription   (SJR: 0.124, h-index: 9)
Angiologia e Cirurgia Vascular     Open Access   (Followers: 1)

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Journal Cover Animal Reproduction Science
  [SJR: 0.711]   [H-I: 78]   [6 followers]  Follow
   Hybrid Journal Hybrid journal (It can contain Open Access articles)
   ISSN (Print) 0378-4320
   Published by Elsevier Homepage  [3175 journals]
  • Synchronization of cyclic and acyclic embryo recipient mares with donor
    • Authors: Ivan V. Oliveira Neto; Igor F. Canisso; Lorenzo G. Segabinazzi; Camila P.F. Dell’Aqua; Marco A. Alvarenga; Frederico O. Papa; Jose A. Dell’Aqua
      Pages: 1 - 9
      Abstract: Publication date: March 2018
      Source:Animal Reproduction Science, Volume 190
      Author(s): Ivan V. Oliveira Neto, Igor F. Canisso, Lorenzo G. Segabinazzi, Camila P.F. Dell’Aqua, Marco A. Alvarenga, Frederico O. Papa, Jose A. Dell’Aqua
      This study compared hormone treatments given to mares during anestrus, spring transition, and different stages of the estrous cycle, by assessing uterine features and pregnancy rates after embryo transfer (ET). Embryo recipient mares (n = 160) were equally arranged as follows: G1-spontaneous ovulation (control), G2-anestrus, G3-spring transition, G4-early estrus, G5-estrus, G6-diestrus, G7-early diestrus treated with a dose of dinoprost, and G8-early diestrus treated with two doses of dinoprost. At treatment initiation (Day-4), G2-7 were given dinoprost and estradiol-17β, thereafter, estradiol-17β was repeated on Days-3,-2, and -1. On Day0, mares received long-acting altrenogest. Then, each mare had one ET performed from Day + 3 to Day + 8 after altrenogest. Immediately before the ET, mares received a boost of altrenogest and had uterine features assessed. Pregnant mares on each of the checks (by 7, 30, 60, and 120d after ET) were maintained on weekly injections of LA-P4 until 120d. G8 received similar management, but dinoprost was repeated on Day-3. G1-G6 and G8 displayed uterine edema and satisfactory pregnancy rates ≥65%. Repeating dinoprost to G8 likely ensured proper luteolysis and response to estrogen as determined by higher uterine edema scores and pregnancy rates than G7 (p < .05). Our results were consistent with previous studies and other successful commercial ET programs (except G7), thus, demonstrating the usefulness of the hormone treatments described herein to synchronize embryo recipient mares with donor mares. Thus, we foresee that other groups may use the strategies described herein for the management of embryo recipient mares.

      PubDate: 2018-02-26T02:57:53Z
      DOI: 10.1016/j.anireprosci.2017.12.016
      Issue No: Vol. 190 (2018)
  • Culture of somatic cells isolated from frozen-thawed equine semen using
           fluorescence-assisted cell sorting
    • Authors: Joao Gatto Brom-de-Luna; Heloísa Siqueira Canesin; Gus Wright; Katrin Hinrichs
      Pages: 10 - 17
      Abstract: Publication date: March 2018
      Source:Animal Reproduction Science, Volume 190
      Author(s): Joao Gatto Brom-de-Luna, Heloísa Siqueira Canesin, Gus Wright, Katrin Hinrichs
      Nuclear transfer using somatic cells from frozen semen (FzSC) would allow cloning of animals for which no other genetic material is available. Horses are one of the few species for which cloning is commercially feasible; despite this, there is no information available on the culture of equine FzSC. After preliminary trials on equine FzSC, recovered by density-gradient centrifugation, resulted in no growth, we hypothesized that sperm in the culture system negatively affected cell proliferation. Therefore, we evaluated culture of FzSC isolated using fluorescence-assisted cell sorting. In Exp. 1, sperm were labeled using antibodies to a sperm-specific antigen, SP17, and unlabeled cells were collected. This resulted in high sperm contamination. In Exp. 2, FzSC were labeled using an anti-MHC class I antibody. This resulted in an essentially pure population of FzSC, 13–25% of which were nucleated. Culture yielded no proliferation in any of nine replicates. In Exp. 3, 5 × 103 viable fresh, cultured horse fibroblasts were added to the frozen-thawed, washed semen, then this suspension was labeled and sorted as for Exp. 2. The enriched population had a mean of five sperm per recovered somatic cell; culture yielded formation of monolayers. In conclusion, an essentially pure population of equine FzSC could be obtained using sorting for presence of MHC class I antigens. No equine FzSC grew in culture; however, the proliferation of fibroblasts subjected to the same processing demonstrated that the labeling and sorting methods, and the presence of few sperm in culture, were compatible with cell viability.

      PubDate: 2018-02-26T02:57:53Z
      DOI: 10.1016/j.anireprosci.2017.12.017
      Issue No: Vol. 190 (2018)
  • Stage-specific expression of DDX4 and c-kit at different developmental
           stages of the porcine testis
    • Authors: Ran Lee; Won-Young Lee; Hyun-Jung Park; Woo-Tae Ha; Jae-Seok Woo; Hak-Jae Chung; Ji-Heon Lee; Kwonho Hong; Hyuk Song
      Pages: 18 - 26
      Abstract: Publication date: March 2018
      Source:Animal Reproduction Science, Volume 190
      Author(s): Ran Lee, Won-Young Lee, Hyun-Jung Park, Woo-Tae Ha, Jae-Seok Woo, Hak-Jae Chung, Ji-Heon Lee, Kwonho Hong, Hyuk Song
      Spermatogenesis begins with spermatogonial stem cells (SSCs), which are located in the basement membrane of the adult testes. Previous studies have described specific biomarkers for undifferentiated porcine spermatogonia or SSCs; however, these markers are not sufficient to understand spermatogenesis at different developmental stages. The objective of this study was characterize the expression of DEAD-Box polypeptide 4 (DDX4, also known as VASA) and tyrosine-protein kinase kit (c-kit), as potential markers of male germ cells in the porcine testis. In porcine testis tissue at prepubertal stages (5, 30, and 60 days), DDX4 and c-kit protein expression was detected in the most undifferentiated spermatogonia, which also express protein gene product 9.5 (PGP9.5). However, in porcine testis tissues from pubertal and postpubertal stages (90, 120, and 150 days), DDX4 and c-kit were not detected in PGP9.5-positive undifferentiated spermatogonia. The DDX4 expression pattern was similar to that of c-kit in the porcine testis. In adult porcine testes, DDX4-expressing cells were located on the lumenal side, compared to synaptonemal complex protein 3-positive primary spermatocytes, but DDX-4 was not co-expressed with acrosin, a known acrosome marker. In addition, DDX4 was detected in PGP9.5-expressing porcine SSCs in culture. Based on our results, we suggest that DDX4 and c-kit are putative markers of undifferentiated spermatogonia in the prepubertal porcine testis. While in the postpubertal porcine testis, they are markers of differentiated spermatocytes. These findings may facilitate future studies of porcine spermatogenesis.

      PubDate: 2018-02-26T02:57:53Z
      DOI: 10.1016/j.anireprosci.2017.12.020
      Issue No: Vol. 190 (2018)
  • Age-associated expression of vitamin D receptor and vitamin D-metabolizing
           enzymes in the male reproductive tract and sperm of Hu sheep
    • Authors: Xiaolei Yao; M.A. EI-Samahy; Hua Yang; Xu Feng; Fengzhe Li; Fanxing Meng; Haitao Nie; Feng Wang
      Pages: 27 - 38
      Abstract: Publication date: Available online 10 January 2018
      Source:Animal Reproduction Science
      Author(s): Xiaolei Yao, M.A. EI-Samahy, Hua Yang, Xu Feng, Fengzhe Li, Fanxing Meng, Haitao Nie, Feng Wang
      The cellular response to 1,25-dihydroxyvitamin D3 (Vit D3; biologically active form of Vitamin D) is complex and depends not only on Vitamin D receptor (VDR) expression but also on cellular uptake of circulating Vit D3 and the presence and activity of Vitamin D-metabolizing enzyme. This study evaluated the expression of VDR and Vitamin D-metabolizing enzymes in the ram reproductive tract at different developmental stages and in spermatozoa. Nearly all cell types in the testes and epithelial cells of the caput, corpus, and cauda expressed VDR, CYP27B1, and CYP24A1 proteins. The mRNA and protein expression of CYP2R1, CYP27A1, and CYP27B1 in the testes and cauda increased significantly with increasing age (P < 0.05). However, epididymal VDR mRNA and protein expression showed no significant difference (P < 0.05) between adult (9- and 24-month-old) and prepubertal (3-month-old) rams. Furthermore, VDR and CYP24A1 were mainly concentrated in the mid-piece of ejaculated or cauda epididymis spermatozoa or both. Additionally, VDR and CYP27B1 mRNA and protein expression levels were significantly higher in ejaculated spermatozoa than in cauda epididymal spermatozoa (P < 0.05). Moreover, VDR and CYP24A1 expression was significantly higher in high-motility than in low-motility spermatozoa (P < 0.05). The diverse expression patterns of VDR and Vitamin D-metabolizing enzymes in the ram reproductive tract at different developmental stages and spermatozoa suggest it plays a potential role in spermatogenesis.

      PubDate: 2018-02-04T19:48:52Z
      DOI: 10.1016/j.anireprosci.2018.01.003
      Issue No: Vol. 190 (2018)
  • Regulation of conceptus interferon-tau gene subtypes expressed in the
           uterus during the peri-implantation period of cattle
    • Authors: Min-Su Kim; Kwan-Sik Min; Hwan-Hoo Seong; Chan-Lan Kim; Ik Soo Jeon; Sung Woo Kim; Kazuhiko Imakawa
      Pages: 39 - 46
      Abstract: Publication date: Available online 3 February 2018
      Source:Animal Reproduction Science
      Author(s): Min-Su Kim, Kwan-Sik Min, Hwan-Hoo Seong, Chan-Lan Kim, Ik Soo Jeon, Sung Woo Kim, Kazuhiko Imakawa
      Conceptus interferon tau (IFNT), produced by the embryonic trophectoderm, is known as a major signaling protein essential for the process of maternal recognition of pregnancy in ruminants. Similar to other IFN gene families such as IFNA and IFNB, multiple IFNT genes exist. The number of IFNT genes actively transcribed and regulated in conceptuses of cattle has, however, not been well characterized. In this study, IFNT transcripts in utero were studied through the use of next generation sequencer. Among 38 IFN genes registered and eight annotated as IFNT, only two transcripts, IFNT1 and IFNTc1, were found in conceptuses in utero. Relative abundance of transcription factor(s) involved in the regulation of IFNT genes were investigated by real-time PCR. Transcriptional activity of IFNT1 and IFNTc1 were investigated using bovine non-trophoblast ear fibroblast (EF) cells, which were co-transfected with luciferase reporter constructs with upstream (−631 to −51) promoter regions of IFNT1 or IFNTc1 and various transcription factor expression plasmids, CDX2, AP1 (JUN), ETS2 and/or CREBBP. CDX2 with AP1 and ETS2 was found to increase luciferase activity of IFNT1 and IFNTc1 approximately 14- and 11-fold, respectively, in EF cells, which do not express the CDX2 gene. These results indicated that two isoforms of conceptus IFNT genes of cattle could be regulated differently in utero. Furthermore, IFNT1 and IFNTc1 were found to have similar antiviral activity, suggesting that both IFNT genes could function to increase conceptus signaling to the uterine endometrium for the process of maternal recognition of pregnancy during the pre-implantation period.

      PubDate: 2018-02-04T19:48:52Z
      DOI: 10.1016/j.anireprosci.2018.01.005
      Issue No: Vol. 190 (2018)
  • Effect of GnRH analogue administration on Day 7 after natural mating on
           formation accessory corpus luteum, progesterone concentration and
           conception rate in llamas (Lama glama)
    • Authors: Marcos C. Abalos; Francisco Acuña; Andrea K. Cancino; Juan F. Aller
      Pages: 47 - 52
      Abstract: Publication date: Available online 31 January 2018
      Source:Animal Reproduction Science
      Author(s): Marcos C. Abalos, Francisco Acuña, Andrea K. Cancino, Juan F. Aller
      The objectives of the present study were to determine the effects of exogenous GnRH administered 7 days after breeding on the formation of an accessory corpus luteum (ACL), plasma progesterone (P4) concentrations and pregnancy rates. Adult females (n = 71) having a follicle ≥ 7 mm in diameter in the ovary were naturally mated (Day 0). On Day 7, ultrasonic examination was performed to confirm the occurrence of ovulation as evidenced by presence of an induced corpus luteum (ICL). Females with an ICL plus a dominant follicle ≥ 7 mm (n = 56) were treated with saline solution (SS, n = 29) or GnRH analogue (n = 27). On Day 14, the formation of an ACL was observed by ultrasonography. Blood samples were collected on Days 7 and 14 to quantify plasma P4 concentrations. On Day 14, 21 of 27 (77.8%) females in the GnRH group developed an ACL, whereas females in the SS group did not. Progesterone concentrations on Day 7 and 14 in those llamas diagnosed as pregnant on Day 30 were not different (P > 0.05) between groups. In addition, P4 concentration was similar for GnRH-treated females having two CL to those with a single CL. Pregnancy rates were similar (P > 0.05) between SS and GnRH groups (55.2% compared with 74.1% respectively) and the pregnancy rate for the GnRH group was not affected (P > 0.05) by the number of CL observed at Day 14 (66.6% and 75.6% for females with one and two CL respectively). In conclusion, GnRH administration on Day 7 after breeding leads to ACL formation; however, neither the plasma P4 concentration nor pregnancy rate was affected by having an ACL.

      PubDate: 2018-02-04T19:48:52Z
      DOI: 10.1016/j.anireprosci.2018.01.006
      Issue No: Vol. 190 (2018)
  • Comparison of 3 anesthetic protocols for the elective cesarean-section in
           the dog: Effects on the bitch and the newborn puppies
    • Authors: José M. Vilar; M. Batista; R. Pérez; A. Zagorskaia; E. Jouanisson; L. Díaz-Bertrana; S. Rosales
      Pages: 53 - 62
      Abstract: Publication date: Available online 1 February 2018
      Source:Animal Reproduction Science
      Author(s): José M. Vilar, M. Batista, R. Pérez, A. Zagorskaia, E. Jouanisson, L. Díaz-Bertrana, S. Rosales
      This study assessed the influence of 3 different anesthetic protocols based on the quality of anesthesia induction and maintenance in four dog breeds (French Bulldog, n = 13; Yorkshire terrier, n = 12; Chihuahua, n = 10; Bull Terrier, n = 10) subjected to cesarean section. Neonatal mortality, birth defects and newborn viability were assessed. All females were pre-medicated with morphine (IM), and then were assigned to three different anesthetic protocols: group P (n = 17), anesthesia was induced with propofol (IV) and then also maintained with propofol until the complete delivery of puppies and then anesthesia was maintained afterwards with sevoflurane; group PS (n = 14), anesthesia was induced with IV propofol, and maintenance of the anesthesic plan was performed with sevoflurane; group PES (n = 14) the females were induced by propofol and an epidural anesthesia was then performed, anesthesia was then maintained with propofol until the complete extraction of all puppies and then anesthesia was maintained afterwards with sevoflurane. Throughout the surgery, group PES required a lower concentration of sevoflurane (p < 0.05), and extra doses of propofol or fentanyl during inhalatory anesthesia were not required. Mean values of heart rate (p < 0.01) were higher in females from groups P and PS. Mean values of blood pressure values were lower (p < 0.01) in group PES as compared with the other two groups. Birth defects were detected in 3.1% (5/162) of the neonates, with a significantly higher incidence (p < 0.05) in French bulldog puppies. Neonatal viability was assessed using a modified Apgar score model; Apgar score was defined immediately after delivery (Apgar0) and a second score was assessed 60 min after delivery (Apgar60). Apgar0 scores were significantly different between the groups, showing neonates of group PES the highest values (p < 0.05). In Apgar60, more than 94% of puppies were already classified as normal viability neonates (7–10 score) and no differences were observed between groups. This study confirmed that females of group PES showed a higher quality of anesthesia during surgery and a vitality of puppies immediately after delivery. Regardless of the anesthetic protocol used, French bulldog females and puppies required more clinical care than other breeds.

      PubDate: 2018-02-04T19:48:52Z
      DOI: 10.1016/j.anireprosci.2018.01.007
      Issue No: Vol. 190 (2018)
  • Changes in external osmolality and ionic composition affect Megaleporinus
           obtusidens sperm motility
    • Authors: Jurandir Joaquim Bernardes Júnior; Jhon Edison Jimenez; Robie Allan Bombardelli; Alex Pires de Oliveira Nuñer
      Pages: 63 - 74
      Abstract: Publication date: Available online 1 February 2018
      Source:Animal Reproduction Science
      Author(s): Jurandir Joaquim Bernardes Júnior, Jhon Edison Jimenez, Robie Allan Bombardelli, Alex Pires de Oliveira Nuñer
      Understanding the effects of environmental factors on sperm motility characteristics can increase artificial reproduction efficiency in species that do not spawn naturally in captivity, such as Megaleporinus obtusidens. This study evaluated the effects of the osmolality (25, 85, 145, 205, 265, and 325 mOsm kg−1) and composition of activating solutions (NaCl, KCl, or fructose) on the percentage of motile sperm, and the swimming velocity and path straightness of M. obtusidens spermatozoa. The concentrations of major ions in the seminal fluid were also assessed and Na+ (74.46 mmol L−1), K+ (37.24 mmol L−1), and Cl− (114.29 mmol L−1) were the most abundant. When the activating solution was hypertonic (325 mOsm kg−1) compared to the seminal fluid (293 mOsm kg−1), sperm motility was completely inhibited. A wide range of osmolalities that initiated sperm motility were identified for all three solutions. Both, the percentage of motile sperm and the motility duration were reduced (P < .05) at extreme osmolalities. At 145 mOsm kg−1, the percentage of motile sperm remained high (>50%) up to 40 s after activation and the motile phase lasted for >50 s, regardless of the activating solution composition. Over the postactivation time, the curvilinear velocity and straightness were similar (P > .05) for fructose and NaCl solutions, whereas KCl solutions induced a higher (P < .05) curvilinear velocity, lower (P < .05) straight-line velocity, and a circular swimming motion in spermatozoa. Our results suggest that a reduction in osmolality, using both non-electrolyte and electrolyte solutions, is the main trigger for the onset of spermatic movement in M. obtusidens sperm.

      PubDate: 2018-02-04T19:48:52Z
      DOI: 10.1016/j.anireprosci.2018.01.008
      Issue No: Vol. 190 (2018)
  • Reproductive responses to sexually active buck of does treated with
           melatonin when body weight/body condition is increasing or decreasing
    • Authors: L. Gallego-Calvo; M.C. Gatica; J.L. Guzmán; L.A. Zarazaga
      Pages: 75 - 84
      Abstract: Publication date: Available online 1 February 2018
      Source:Animal Reproduction Science
      Author(s): L. Gallego-Calvo, M.C. Gatica, J.L. Guzmán, L.A. Zarazaga
      When the sexual activity of bucks is minimal, there is a minimal male effect on does regardless of their body weight (BW)/body condition (BC) and whether does are treated with melatonin or not. The study examines whether sexually active bucks can induce an adequate male effect in does with an increasing or decreasing trajectory of change in BW/BC when does are or not treated with melatonin. During natural anoestrus, 46 Blanca Andaluza does were assigned to two groups: 1) low BW/low BC group in which does were fed 1.9 times maintenance requirements for dietary energy for gaining BW/BC (LLg group; n = 23); or 2) a high BW/high BC group in which the does were fed 0.4 times maintenance requirements for dietary energy that resulted in a loss of BW/BC (HHl group; n = 23). There were similar numbers of does in each group that were treated or not treated with melatonin (MEL). Following 48 days of isolation from bucks, four sexually active individuals fitted with marking harnesses were transferred to the paddock containing the does of each group. Blood samples were collected by jugular venipuncture (before the distribution of concentrate) twice per week. The effect of the treatments (increasing or decreasing BW/BC and melatonin) on the different variables that were assessed were analysed using an ANOVA or the Fisher-Freeman-Halton exact probability test as necessary. During the 35 days after treatments were applied, the percentage of females expressing oestrous and having an ovulation were greater in the LLg + MEL than HHl-MEL subgroup (P < 0.05). The interaction of nutrition × melatonin treatment had a significant effect on reproduction of does (P < 0.05). This could be explained by the greater plasma glucose and IGF-1 and lesser plasma non-esterified fatty acid concentrations in does with increasing BW/BC (P < 0.01), and the greater IGF-1 concentrations of MEL-treated females (P < 0.01). The LH concentration and pulsatile release of this hormone from the pituitary were also modified by the presence of the males (P < 0.01). Furthermore, the LLg + MEL-treated does were responsive to the presence of bucks (P < 0.05). The present results indicate sexually active males cannot induce an adequate reproductive response in females with decreasing BW/BC even when does are being treated with melatonin. The presence of bucks enhanced the doe reproductive response when does were treated with melatonin and a pattern of increasing BW/BC.

      PubDate: 2018-02-04T19:48:52Z
      DOI: 10.1016/j.anireprosci.2018.01.009
      Issue No: Vol. 190 (2018)
  • Freeze-dried spermatozoa: an alternative biobanking option for endangered
    • Authors: Debora Agata Anzalone; Luca Palazzese; Domenico Iuso; Giuseppe Martino; Pasqualino Loi
      Pages: 85 - 93
      Abstract: Publication date: Available online 31 January 2018
      Source:Animal Reproduction Science
      Author(s): Debora Agata Anzalone, Luca Palazzese, Domenico Iuso, Giuseppe Martino, Pasqualino Loi
      In addition to the iconic wild species, such as the pandas and Siberian tigers, an ever-increasing number of domestic species are also threatened with extinction. Biobanking of spermatozoa could preserve genetic heritages of extinct species, and maintain biodiversity of existing species. Because lyophilized spermatozoa retain fertilizing capacity, the aim was to assess whether freeze-dried spermatozoa are an alternative option to save endangered sheep breeds. To achieve this objective, semen was collected from an Italian endangered sheep breed (Pagliarola), and a biobank of cryopreserved and freeze-dried spermatozoa was established, and evaluated using IVF (for frozen spermatozoa) and ICSI procedures (for frozen and freeze-dried spermatozoa). As expected, the fertilizing capacity of cryopreserved Pagliarola’s spermatozoa was comparable to commercial semen stocks. To evaluate the activating capability of freeze-dried spermatozoa, 108 MII sheep oocytes were subjected to ICSI, and allocated to two groups: 56 oocytes were activated by incubation with ionomycin (ICSI-FDSa) and 52 were not activated (ICSI-FDSna). Pronuclear formation (2PN) was investigated at 14 to 16 hours after ICSI in fixed presumptive zygotes. Only artificially activated oocytes developed into blastocysts after ICSI. In the present study, freeze-dried ram spermatozoa induced blastocyst development following ICSI at a relatively high proportion, providing evidence that sperm lyophilization is an alternative, low cost storage option for biodiversity preservation of domestic species.

      PubDate: 2018-02-04T19:48:52Z
      DOI: 10.1016/j.anireprosci.2018.01.010
      Issue No: Vol. 190 (2018)
  • Effect of production management on semen quality during long-term storage
           in different European boar studs
    • Authors: M. Schulze; C. Kuster; J. Schäfer; M. Jung; R. Grossfeld
      Pages: 94 - 101
      Abstract: Publication date: Available online 31 January 2018
      Source:Animal Reproduction Science
      Author(s): M. Schulze, C. Kuster, J. Schäfer, M. Jung, R. Grossfeld
      The processing of ejaculates is a fundamental step for the fertilizing capacity of boar spermatozoa. The aim of the present study was to identify factors that affect quality of boar semen doses. The production process during 1 day of semen processing in 26 European boar studs was monitored. In each boar stud, nine to 19 randomly selected ejaculates from 372 Pietrain boars were analyzed for sperm motility, acrosome and plasma membrane integrity, mitochondrial activity and thermo-resistance (TRT). Each ejaculate was monitored for production time and temperature for each step in semen processing using the special programmed software SEQU (version 1.7, Minitüb, Tiefenbach, Germany). The dilution of ejaculates with a short-term extender was completed in one step in 10 AI centers (n = 135 ejaculates), in two steps in 11 AI centers (n = 158 ejaculates) and in three steps in five AI centers (n = 79 ejaculates). Results indicated there was a greater semen quality with one-step isothermal dilution compared with the multi-step dilution of AI semen doses (total motility TRT d7: 71.1 ± 19.2%, 64.6 ± 20.0%, 47.1 ± 27.1%; one-step compared with two-step compared with the three-step dilution; P < .05). There was a marked advantage when using the one-step isothermal dilution regarding time management, preservation suitability, stability and stress resistance. One-step dilution caused significant lower holding times of raw ejaculates and reduced the possible risk of making mistakes due to a lower number of processing steps. These results lead to refined recommendations for boar semen processing.

      PubDate: 2018-02-04T19:48:52Z
      DOI: 10.1016/j.anireprosci.2018.01.011
      Issue No: Vol. 190 (2018)
  • Effect of luteinizing hormone on goat theca cell apoptosis and
           steroidogenesis through activation of the PI3K/AKT pathway
    • Authors: Xiaomei Wang; Pengda Zou; Yuanyuan He; Kai Meng; Fusheng Quan; Yong Zhang
      Pages: 108 - 118
      Abstract: Publication date: Available online 1 February 2018
      Source:Animal Reproduction Science
      Author(s): Xiaomei Wang, Pengda Zou, Yuanyuan He, Kai Meng, Fusheng Quan, Yong Zhang
      Luteinizing hormone (LH) is a glycoprotein that regulates the function of ovarian follicular cells. Theca cells (TCs) also have a key role in follicular growth and atresia. The effects and intracellular signaling mechanisms were investigated of LH on apoptosis and steroidogenesis in goat gonadotropin-independent follicular (1.0–4.0 mm) TCs. The results indicated that LH increased androstenedione secretion and relative abundance of CYP17A1 and BCL2 mRNA in the TCs, whereas LH in combination with LY294002, a PI3K/AKT inhibitor, decreased LH-induced function. The apoptosis ratio and expression of the BAX gene in TCs were less with LH treatment, and the extent of this inhibition was decreased by suppressing the PI3K/AKT pathway. In conclusion, results of the present study indicate LH regulates apoptosis and steroidogenesis in goat TCs by activating the PI3K/AKT pathway.

      PubDate: 2018-02-04T19:48:52Z
      DOI: 10.1016/j.anireprosci.2018.01.014
      Issue No: Vol. 190 (2018)
  • Endometrial blood perfusion as assessed using a novel laser Doppler
           technique in Angus cows
    • Authors: M.P.T. Owen; K.J. McCarty; C.G. Hart; C.S. Steadman; C.O. Lemley
      Pages: 119 - 126
      Abstract: Publication date: Available online 1 February 2018
      Source:Animal Reproduction Science
      Author(s): M.P.T. Owen, K.J. McCarty, C.G. Hart, C.S. Steadman, C.O. Lemley
      Previous studies have characterized ovarian steroid synthesis which directly affects uterine environment and blood flow. Clearance of steroids occurs primarily in hepatic tissues, however, it was discovered that there is an abundant activity of the phase II steroid metabolizing enzyme UDP-glucuronosyltransferase (UGT) in uterine biopsies. No minimally invasive techniques for collecting endometrial perfusion, which is affected by steroids and indicative of reproductive health, have been developed for livestock. The objective of the present study was to characterize UGT activity and endometrial blood perfusion during a normal estrous cycle of cattle. It was hypothesized that there would be increased steroid metabolism during the luteal phase of the estrous cycle and in the uterine horn ipsilateral to the corpus luteum (CL). During the first synchronized estrous cycle, progesterone and UGT activity increased on Day 6 compared with 0 and 3, with the first day of estrus being considered Day 0 of the study. Endometrial perfusion was greater ipsilateral to the CL compared with contralateral on Day 12, and was less ipsilateral to the CL compared with contralateral on Day 18. Similar to perfusion results, nitric oxide metabolites (nitrites) were greatest in the endometrium ipsilateral as compared with contralateral to the CL. Moreover, there was a positive correlation (r = 0.28; P = 0.04) between endometrial perfusion and nitrite concentration. It is concluded that activity of UGT within the endometrium is affected by the contralateral or ipsilateral location of the CL, and collection of endometrial perfusion data using a laser Doppler probe could be a viable measurement technique as indicated by associated nitrite concentrations in the present study.

      PubDate: 2018-02-04T19:48:52Z
      DOI: 10.1016/j.anireprosci.2018.01.015
      Issue No: Vol. 190 (2018)
  • Purification, structural and biophysical characterisation of the major
           seminal plasma protein from Texel rams
    • Authors: Manoel Augusto Klempovus Villela Condessa; Arethusa Lobo Pimentel; Flávio Augusto Vicente Seixas; Antonio Campanha Martinez
      Pages: 11 - 18
      Abstract: Publication date: February 2018
      Source:Animal Reproduction Science, Volume 189
      Author(s): Manoel Augusto Klempovus Villela Condessa, Arethusa Lobo Pimentel, Flávio Augusto Vicente Seixas, Antonio Campanha Martinez
      Spermadhesins are a group of low molecular weight proteins present in seminal plasma. In Texel rams, they represent more than 70% of the seminal plasma proteins. Although their functions have not yet been fully clarified, there is much discussion about the role of these proteins in maintaining sperm viability during and after the semen freezing process. This work sought to isolate the major component of the seminal plasma from rams of the Texel breed (O. aries SPD2) and to evaluate its structural and biophysical characteristics in order to better understand its role in spermatic viability. The protein was isolated by centrifugation and ion exchange chromatography and its biophysical properties were evaluated by circular dichroism spectrometry. Molecular dynamics simulations of the modelled protein compared to the homologous bovine protein were also carried out. The results showed that O. aries SPD2 has a transition temperature (Tm ) of 65 °C and a ΔHm of 322.5 kJ mol−1, similar to the results for other spermadhesins described in the literature. The estimated composition of the secondary structure elements for the native protein is in agreement with that observed for the theoretical model. Its structural characteristics were preserved in simulations at temperatures of 27 °C and 40 °C, as was the case for bull spermadhesin. Taken together, these results suggest that the major component of the spermadhesins of Texel rams (O. aries SPD2) may play an important role in maintaining the viability of spermatozoa in fresh semen as well as after thawing.

      PubDate: 2018-02-04T19:48:52Z
      DOI: 10.1016/j.anireprosci.2017.10.013
      Issue No: Vol. 189 (2018)
  • Effects of coenzyme Q10 on semen cryopreservation of stallions classified
           as having good or bad semen freezing ability
    • Authors: Joao A.M. Carneiro; Igor F. Canisso; R.S. Bandeira; V.F.C. Scheeren; Camila P. Freitas-Dell’Aqua; Marco A. Alvarenga; Frederico O. Papa; Jose A. Dell’Aqua
      Abstract: Publication date: Available online 24 February 2018
      Source:Animal Reproduction Science
      Author(s): Joao A.M. Carneiro, Igor F. Canisso, R.S. Bandeira, V.F.C. Scheeren, Camila P. Freitas-Dell’Aqua, Marco A. Alvarenga, Frederico O. Papa, Jose A. Dell’Aqua
      This study aimed to evaluate the antioxidant properties of coenzyme Q10 (CoQ10) during cryopreservation of semen obtained from stallions having good and bad semen freezing ability (GFA vs. BFA, respectively). Forty ejaculates (n=20 stallions) were split into five centrifugation and five freezing extenders containing different concentrations of CoQ10 (0, 25, 50, 75 and 100 µmols/L). If CoQ10 was added to the centrifugation extender, the freezing extender had no CoQ10 added; similarly, if CoQ10 was added to the freezing extender, the centrifugation extender had no CoQ10. Semen cryopreserved on extenders containing no CoQ10 served as the control. After post-thaw total sperm motility (TM) assessments, the stallions were classified as GFA (i.e., decrease of ≤25% in TM, n=7) or BFA (i.e., decrease of ≥ 40% in TM, n=5). Stallions not fitting (n=8) this enrollment criteria had samples discarded. After that, two straws for each extender were thawed at 37ºC for 30s; one straw was immediately used for evaluation of sperm kinetics, plasma membrane integrity, non-capacitated spermatozoa, reactive oxygen species production, mitochondrial activity and lipid peroxidation. The second straw was kept at 37ºC for 30 min and subjected to the same assessments. Expectedly, sperm motility parameters were significantly lower for stallions with BFA. There were no effects of CoQ10 concentration or time for all parameters evaluated in the group with GFA when compared with the control extender (p>0.05), except lipid peroxidation (p <0.05). However, stallions with BFA had improved sperm parameters for samples processed with extenders containing CoQ10 (particularly 75 µmols/L) (p <0.05), except for the reactive oxygen species production and mitochondrial potential (T0) in which there were no differences between the groups (p> 0.05). In summary, 75 µmols/L appears to be the optimal dose of Co-Q10, particularly, when added to the centrifugation extender.

      PubDate: 2018-02-26T02:57:53Z
      DOI: 10.1016/j.anireprosci.2018.02.020
  • Seminal plasma affects the survival rate and motility pattern of raw llama
    • Authors: F.G. Fumuso; S.M. Giuliano; M.G. Chaves; D.M. Neild; M.H. Miragaya; M.C. Gambarotta; M.I. Carretero
      Abstract: Publication date: Available online 24 February 2018
      Source:Animal Reproduction Science
      Author(s): F.G. Fumuso, S.M. Giuliano, M.G. Chaves, D.M. Neild, M.H. Miragaya, M.C. Gambarotta, M.I. Carretero
      The objectives of this study were to evaluate the effect over time of different percentages of seminal plasma (SP) on llama sperm characteristics in raw semen and correlate the techniques routinely used to evaluate sperm viability and acrosome status with the Fluorescein Isothiocyanate –Arachis hypogea agglutinin/Propidium Iodide (FITC-PNA/PI). Eighteen ejaculates, obtained from 6 male llamas using electroejaculation, were incubated in 0.1% collagenase in HEPES-TALP (HT), centrifuged and resuspended with SP and HT: 0, 10, 50 and 100% SP. Samples were incubated (37 °C) until evaluation at 0; 1.5 and 3 h. Split plot and factorial designs were used to analyze sperm motility, viability, membrane function and acrosome status and Spearman’s test was used for correlation. At 0 h, samples with 100% SP showed oscillatory motility; whereas in samples with 0 and 10% SP, progressive motility was predominant. Viability, membrane function and total motility decreased significantly at 3 h of incubation in samples with 100% SP. Sperm with intact acrosomes were fewer in 0% SP media at all times. FITC-PNA/PI correlated with 6-Carboxyfluorescein Diacetate and Propidium Iodide (CFDA/PI) and with Coomassie Blue (CB) stains (r=0.8; p=0.0 and r=0.5; p=0.0 respectively). Conclusions: the motility pattern of llama sperm is influenced by the concentration of SP. The use of SP as the only medium is not able to maintain sperm motility, viability and membrane function for 3 h. A certain percentage of SP is necessary in the medium to avoid spontaneous acrosome reactions. The correlations observed could help to shorten evaluation times and reduce costs in sperm laboratories.

      PubDate: 2018-02-26T02:57:53Z
      DOI: 10.1016/j.anireprosci.2018.02.019
  • Melatonin mitigates bisphenol A-induced estradiol production and
           proliferation by porcine ovarian granulosa cells in vitro
    • Authors: Guoyun Wu; Dan Song; Quanwei Wei; Jun Xing; Xiaoli Shi; Fangxiong Shi
      Abstract: Publication date: Available online 24 February 2018
      Source:Animal Reproduction Science
      Author(s): Guoyun Wu, Dan Song, Quanwei Wei, Jun Xing, Xiaoli Shi, Fangxiong Shi
      Melatonin plays a crucial role in the amelioration of reproductive toxicity induced by endocrine-disrupting chemicals. However, very few studies have investigated the mitigating effects of melatonin on BPA-induced dysfunction in porcine granulosa cells. In the present study, primary granulosa cells were cultured in serum-low conditions with bisphenol A (BPA) (10 μM) with or without melatonin (100 μM), followed by evaluation of estradiol synthesis and cell proliferation. Our results showed that BPA significantly increased estradiol concentration and granulosa cell proliferation. Interestingly, melatonin co-incubation reduced the high levels of estradiol in porcine ovarian granulosa cells induced by BPA stimulation. Furthermore, melatonin co-incubation also attenuated BPA-induced proliferation as shown by a decline in the Ki67-positive cell ratio and PCNA expression level. However, treatment with melatonin-alone did not dramatically reduce estradiol levels or expression of proliferative regulatory protein markers (Ki67, PCNA). We hypothesize that the regulation by melatonin of estradiol biosynthesis and cellular proliferation is highly correlated with BPA stimulation. In conclusion, this study first showed that melatonin mitigated BPA-induced estradiol increase and proliferation in porcine ovarian granulosa cells in vitro. Our results suggest that melatonin may be a promising pharmacologic agent for preventing the potential reproductive toxicity caused by endocrine-disrupting chemicals.

      PubDate: 2018-02-26T02:57:53Z
      DOI: 10.1016/j.anireprosci.2018.02.018
  • In utero heat stress causes reduced testicular area at puberty, reduced
           total sperm production, and increased sperm abnormalities in boars
    • Authors: Drew W. Lugar; Jarret A. Proctor; Timothy J. Safranski; Matthew C. Lucy; Kara R. Stewart
      Abstract: Publication date: Available online 24 February 2018
      Source:Animal Reproduction Science
      Author(s): Drew W. Lugar, Jarret A. Proctor, Timothy J. Safranski, Matthew C. Lucy, Kara R. Stewart
      In utero stress has been shown to negatively affect intact male rats and mice, though very little research has been conducted in boars. The objectives of the present studies were to determine the effects of in utero heat stress (IUHS) on postnatal development and the response to postnatal heat stress of boars. Ten boars were selected at weaning from litters subjected to IUHS or in utero thermoneutral (IUTN) during 30–60 days of gestation. The boars were evaluated for reproductive performance from birth through 57 weeks of age (WOA). Testicular area tended to be smaller for IUHS boars compared to IUTN boars at 24 WOA (P = 0.080). Libido did not differ for IUHS or IUTN (P = 0.823). Total sperm production was reduced in IUHS boars compared to IUTN boars (P ≤ 0.040). Semen volume and semen concentration did not differ (P ≥ 0.447 and P ≥ 0.586, respectively). Total motility and progressive motility did not differ for IUHS and IUTN boars (P ≥ 0.359 and P ≥ 0.461, respectively). In utero heat stressed boars had a greater incidence of sperm with tail abnormalities than IUTN (P ≤ 0.029). In utero heat stressed boars had a lower incidence of sperm with proximal droplets following mild, postnatal heat stress compared with IUTN (P = 0.005). In utero heat stress resulted in significant reductions in sperm production and increased sperm abnormalities in boars. The IUHS boars may be slightly more tolerant to postnatal heat stress, though more research is needed.

      PubDate: 2018-02-26T02:57:53Z
      DOI: 10.1016/j.anireprosci.2018.02.022
  • Reproductive parameters of donkey jacks undergoing puberty
    • Authors: Alessandra Rota; Battista Puddu; Chiara Sabatini; Duccio Panzani; Anne-Lyse Lainé; Francesco Camillo
      Abstract: Publication date: Available online 24 February 2018
      Source:Animal Reproduction Science
      Author(s): Alessandra Rota, Battista Puddu, Chiara Sabatini, Duccio Panzani, Anne-Lyse Lainé, Francesco Camillo
      In male donkeys, puberty and the related events have been poorly characterized. The aim of this study was to evaluate the age at which male donkeys reach puberty, and characterize age associated changes in testicular size, testicular blood flow, serum testosterone concentration and semen quality. Every two months, starting at 6 months and finishing at 24 months of age, five male donkeys born in May to July were subjected to B-mode ultrasound examination to assess testicular size and scrotum content and blood serum sampling for testosterone concentration. From the age of 8 months, pulsed Doppler was employed to evaluate blood flow in the testicular artery. Testosterone serum concentration was evaluated via RIA. From the age of 12 months, monthly semen collections were attempted and semen was evaluated for sperm number, motility and morphology. Onset of puberty was defined as the first ejaculate containing ≥50x106 spermatozoa with ≥10% total motility. One of the donkeys was excluded from the statistical analyses due to a hydrocele presented during the study. Testes width was affected by age (P<0.0001) and after an initial plateau increased linearly from 10 months of age. Pulsatility and resistivity indexes were also affected by age (P<0.01), being significantly higher at 14 months than at 24 months. Testosterone serum concentration was affected by age (P<0.0001) and was significantly lower at 6 months (0.1 ng/ml) compared to 22-24 months (≥0.8 ng/ml). Spermatozoa appeared in the ejaculate at a mean age of 18.7 months and puberty was attained between 19 and 20 months of age (mean:19.5 months), between January and February. In conclusion, late spring born Amiata donkey colts reached puberty at 19-20 months of age. Puberty was accompanied by changes in testicular size, testicular blood flow and serum testosterone concentration.

      PubDate: 2018-02-26T02:57:53Z
      DOI: 10.1016/j.anireprosci.2018.02.021
  • Semen evaluation and in vivo fertility in a Northern Italian pig farm: can
           advanced statistical approaches compensate for low sample size' An
           observational study
    • Authors: Alberto Elmi; Federico Banchelli; Francesca Barone; Paolo Fantinati; Domenico Ventrella; Monica Forni; Maria Laura Bacci
      Abstract: Publication date: Available online 23 February 2018
      Source:Animal Reproduction Science
      Author(s): Alberto Elmi, Federico Banchelli, Francesca Barone, Paolo Fantinati, Domenico Ventrella, Monica Forni, Maria Laura Bacci
      The evaluation of sperm functionality and morphology allows discerning between high and low quality ejaculates, but does not give detailed predictive information regarding in vivo fertility. The current developments in statistical modeling have helped in carrying out reproductive studies, but their biggest limitation is in the size of the dataset to be used. The aim of the present observational study was to evaluate whether advanced statistical approaches, such as mixed effects regression models and bootstrap resampling, can help in assessing the predictive ability of semen parameters in terms of in vivo fertility (farrowing rate and litter size), on a small/medium farm with a limited number of animals. Data regarding 33 ejaculates, including viability, subjective motility and acrosome reaction, were collected. Two hundred and thirty-five sows were inseminated with an outcome of 167 deliveries and 1734 newborn piglets. In order to evaluate the relationships among the parameters measured and fertility, mixed effects regression statistical models were used. Once the covariates to be included in the final models were identified, non-parametric bootstrapping was used. The results showed that the farrowing rate was highly associated with the total number of spermatozoa and subjective motility, while litter size was associated with percentage of acrosome reaction. In conclusion, the proposed statistical approach seemed to be suitable for studies regarding reproduction and fertility, even for relatively small sample sizes. Nonetheless, larger data sets are still preferable and required in order to achieve higher reliability.

      PubDate: 2018-02-26T02:57:53Z
      DOI: 10.1016/j.anireprosci.2018.02.014
  • Comparative effects of zinc oxide, zinc oxide nanoparticle and
           zinc-methionine on hatchability and reproductive variables in male
           Japanese quail
    • Authors: Zeinab Khoobbakht; Mehrdad Mohammadi; Mohammad Roostaei- Ali Mehr; Fahimeh Mohammadghasemi; Mohammad Mehdi Sohani
      Abstract: Publication date: Available online 23 February 2018
      Source:Animal Reproduction Science
      Author(s): Zeinab Khoobbakht, Mehrdad Mohammadi, Mohammad Roostaei- Ali Mehr, Fahimeh Mohammadghasemi, Mohammad Mehdi Sohani
      The objective was to examine the effect of different dietary zinc sources on reproduction of male Japanese quail. A total of 512 quail chicks (day-old) were divided into four groups with four replications for a period of 42 days. After this period, excess chicks were removed to attain the ratio of one male to three females and 16 quail in each subgroup. At 52 to 60 d of age, the eggs were collected and incubated. The basal diet (control) contained no zinc and the other three experimental diets were supplemented with 25 and 50 mg/kg zinc from zinc oxide (ZnO), zinc oxide nanoparticles (ZnONP) and zinc-methionine (Zn-Met) for 1 to 35 and 36 to 60 days, respectively. On day 42, two males from each replicate were euthanized. Males from the ZnO and Zn-Met treatments had an increase (P < 0.05) in seminiferous tubule diameters (STD) and germinal epithelium thickness (GET) compared with the control and ZnONP treatments. Cloacal gland index (CGI) was greatest (P < 0.05) for the Zn-Met compared with the other groups. Testosterone concentration was greater (P < 0.05) in the ZnO and Zn-Met compared with the other groups. Addition of Zn-Met to the diet enhanced (P < 0.05) fertility, hatchability and hatched chick weight compared with the other groups. Early and late embryonic death was greater (P < 0.05) in the control and ZnONP groups, respectively, compared with the other groups. This study indicated that supplementing diets with the Zn-Met source improves male Japanese quail reproductive performance and hatchability traits while zinc oxide nanoparticles have detrimental effects on male Japanese quail reproduction and reduces hatchability.

      PubDate: 2018-02-26T02:57:53Z
      DOI: 10.1016/j.anireprosci.2018.02.017
  • Stallion sperm freezing with sucrose extenders: A strategy to avoid
           permeable cryoprotectants
    • Authors: C. Consuegra; F. Crespo; M. Bottrel; I. Ortiz; J. Dorado; M. Diaz-Jimenez; B. Pereira; M. Hidalgo
      Abstract: Publication date: Available online 21 February 2018
      Source:Animal Reproduction Science
      Author(s): C. Consuegra, F. Crespo, M. Bottrel, I. Ortiz, J. Dorado, M. Diaz-Jimenez, B. Pereira, M. Hidalgo
      The aim of this study was to assess different concentrations of sucrose-based extenders combined with bovine serum albumin (BSA) as an alternative to stallion sperm cryopreservation with permeable cryoprotectants. Semen samples (n = 16) were collected from six stallions. Sperm was cooled, filled in 0.5 mL straws and frozen in nitrogen vapor. Post-thaw sperm kinetic parameters, plasma and acrosome membrane integrity were statistically compared among treatments. In Experiment 1, extenders containing 1% of BSA and different concentrations of sucrose (mmol/L, M): 0, 50, 100, 250, 350 and 450 mM were compared. Use of sucrose [100 mM (S2)] resulted in greater values for most of the sperm kinetic parameters assessed (P < 0.001). There were no differences for plasma membrane integrity, except for when sucrose was used at 50 and 250 mM concentrations, and plasma membrane integrity was less (P < 0.05) when these concentrations were used than with the other sucrose concentrations. In Experiment 2, the selected sucrose extender (S2) was compared to an extender containing glycerol as permeable cryoprotectant. Use of the S2 extender resulted in a lesser proportion of sperm with denuded-acrosomes (P < 0.05) in comparison to use of glycerol and values for several kinetic parameters were also greater (P < 0.05) with use of S2. There were no significant differences for the other parameters assessed in this study. In conclusion, stallion sperm can be frozen in the absence of permeable cryoprotectants, using a combination of sucrose 100 mM with BSA-1% as alternative agents.

      PubDate: 2018-02-26T02:57:53Z
      DOI: 10.1016/j.anireprosci.2018.02.013
  • Climatic factors affecting quantity and quality grade of in vivo derived
           embryos of cattle
    • Authors: Josué Chinchilla-Vargas; Marianna M. Jahnke; Tyler M. Dohlman; Max F. Rothschild; Patrick J. Gunn
      Abstract: Publication date: Available online 17 February 2018
      Source:Animal Reproduction Science
      Author(s): Josué Chinchilla-Vargas, Marianna M. Jahnke, Tyler M. Dohlman, Max F. Rothschild, Patrick J. Gunn
      The present study investigated the effects of climatic variables on the quality grade and quantity of in vivo derived cattle embryos in the Midwestern United States. Climatic information included greatest and least daily temperature, average daily wind speed and average temperature-humidity index for each of the 765 records. The response variables included the number of ovarian structures, viable embryos, quality grade 1 embryos, quality grade 2 embryos, quality grade 3 embryos, freezable embryos (sum of quality grade 1 and quality grade 2 embryos), transferrable embryos (sum of quality grade 1, 2, and 3 embryos), degenerate embryos and unfertilized ova. Measures for variables among the breeds of donors and sires grouped by geographical origin were compared. A negative effect of greater temperatures during the early embryonic development stage tended (P < 0.10) to be associated with a decrease the quality of embryos recovered. Interestingly, the greater the Temperature-Humidity Index (THI) during the early ovarian antral follicular development stage 40 to 45 days prior to ovulation was associated with a tendency for greater numbers of total number of freezable and transferrable embryos recovered per uterine flushing (P < 0.10). Increased wind speed at the early antral follicular phase 40 to 45 days prior to ovulation was associated with an increase in the percentage of quality grade 1 embryos recovered (P < 0.05). Wind speed during the estrous synchronization period was also associated with a lesser number of embryos recovered (P < 0.05). This retrospective study confirms that climatic variables have significant effects on the in vivo production of cattle embryos and that wind speed should be considered in future analyses of factors affecting embryo quality.

      PubDate: 2018-02-26T02:57:53Z
      DOI: 10.1016/j.anireprosci.2018.02.012
  • Comparison of plant- and egg yolk-based semen diluents on in vitro sperm
           kinematics and in vivo fertility of frozen-thawed bull semen
    • Authors: E.M. Murphy; C. O’Meara; B. Eivers; P. Lonergan; S. Fair
      Abstract: Publication date: Available online 15 February 2018
      Source:Animal Reproduction Science
      Author(s): E.M. Murphy, C. O’Meara, B. Eivers, P. Lonergan, S. Fair
      Diluents using components of plant origin have been developed as an alternative to animal based extenders for the dilution of bull semen, however, it is unclear if use of these diluents results in in vivo fertility rates similar to those that occur with use of traditional egg yolk-based diluents. The aim of this study was to assess the effect of semen diluent on 60-day non-return rate (NRR) following artificial insemination (AI) with frozen-thawed bull semen. The effect of semen dilution in one of three different commercial diluents (BullXcell – egg yolk-based, OptiXcell – plant-based or AndroMed – plant-based) on post-thaw total and progressive motility as well as kinematic parameters (Experiment 1) and field fertility (Experiment 2, n = 1,480 inseminations) was assessed. Semen stored in OptiXcell had greater post-thaw total and progressive motility than AndroMed (P < 0.05) but did not differ from BullXcell. Semen stored in BullXcell had a greater beat cross frequency and straight line velocity compared to semen stored in AndroMed (P < 0.05) but did not differ when compared with use of OptiXcell; while values for these variables when using OptiXcell and AndroMed did not differ from each other (P > 0.05). There was no difference in any other sperm kinematic parameters (P > 0.05). There was no effect of diluent on 60-day NRR (71.5%, 67.8% and 70.6% for BullXcell, OptiXcell and AndroMed, respectively). In conclusion, while diluent significantly affected post-thaw sperm motility and kinematics, no effect on 60-day NRR was observed. Given that OptiXcell and AndroMed are animal protein-free media these diluents may be a suitable alternative to BullXcell for the storage of frozen-thawed bull semen.

      PubDate: 2018-02-26T02:57:53Z
      DOI: 10.1016/j.anireprosci.2018.02.010
  • Epididymal sperm from Spix’s yellow-toothed cavies sperm successfully
           cryopreserved in Tris extender with 6% glycerol and 20% egg yolk
    • Authors: Andréia M. Silva; Erica C.G. Praxedes; Lívia B. Campos; Luana G.P. Bezerra; Samara S.J. Moreira; Keilla M. Maia; Ana L.P. Souza; Alexandre R. Silva
      Abstract: Publication date: Available online 15 February 2018
      Source:Animal Reproduction Science
      Author(s): Andréia M. Silva, Erica C.G. Praxedes, Lívia B. Campos, Luana G.P. Bezerra, Samara S.J. Moreira, Keilla M. Maia, Ana L.P. Souza, Alexandre R. Silva
      As a non-threatened hystricognath rodent species, Spix’s yellow-toothed cavies can be used as a model for the development of assisted reproductive techniques for the conservation of closely related species. The objective was to establish a functional protocol for cryopreservation of epididymal sperm from these cavies. Twelve sexually mature males, ∼2 y old and weighing ∼300 g, were euthanized. Sperm were recovered by retrograde flushing of the vas deferens and cauda epididymis with Tris extender. Thereafter, sperm were extended in Tris plus 20% egg yolk, with 3%, 6% or 9% glycerol or dimethyl sulfoxide (DMSO), placed in 0.25 mL straws and cryopreserved in liquid nitrogen. Sperm concentration, motility (using computer-assisted sperm analysis; CASA), plasma membrane integrity, osmotic response, morphology and sperm binding-ability were determined in fresh and frozen-thawed sperm. For most sperm endpoints, glycerol was a more desirable cryoprotectant than DMSO. Data (mean ± SEM) were similar with use of 3%, 6%, and 9% glycerol (P > 0.05) in osmotic response (40.66 ± 6.3%, 42.5 ± 7.1%, and 39.5 ± 5.0% respectably), and membrane integrity (55.17 ± 5.5%, 68.4 ± 4.1%, and 59.1 ± 4.9% respectably). Among concentrations assessed, the use of 6% glycerol resulted in the greatest (P < 0.05) post-thaw values for total motility (60.9 ± 4.4%), rapid subpopulation motility (27.7 ± 3.1%) and sperm-binding capability (227.0 ± 20.2). In conclusion, epididymal sperm from the Spix’s yellow-toothed cavies (G. spixii) are optimally cryopreserved in Tris extender with 6% glycerol and 20% egg yolk.

      PubDate: 2018-02-26T02:57:53Z
      DOI: 10.1016/j.anireprosci.2018.02.009
  • Effects of L-glutamine on boar sperm quality during liquid storage at
    • Authors: Shunwei Wang; Meng Sun; Na Wang; Kang Yang; Haitao Guo; Jingran Wang; Yuting Zhang; Shunli Yue; Jiabo Zhou
      Abstract: Publication date: Available online 15 February 2018
      Source:Animal Reproduction Science
      Author(s): Shunwei Wang, Meng Sun, Na Wang, Kang Yang, Haitao Guo, Jingran Wang, Yuting Zhang, Shunli Yue, Jiabo Zhou
      The quality of boar spermatozoa is affected by oxidative stress during preservation in vitro. It has been demonstrated that L-Glutamine (Gln) can effectively protect cells from oxidative stress-induced injury. There are, however, no reports to date evaluating the effects of Gln on boar semen liquid preservation at 17 °C. The aims of the present study were to elucidate whether the addition of Gln to the extender BTS could improve the quality of boar spermatozoa when stored at 17 °C and to determine the mechanism underlying Gln protection of spermatozoa against preservation-induced damage. Boar semen samples were collected and diluted with Beltsville Thawing Solution (BTS) containing different concentrations (0, 10, 20, 40 or 80 mM) of Gln. The results indicated the addition of 20 mM Gln to the BTS improved (P < 0.05) the motility, acrosome integrity and membrane integrity of boar sperm during liquid preservation. Interestingly, treatment of spermatozoa with Gln addition to the extender resulted in ROS quenching, while enhancing γ-glutamyl cysteine synthetase (γ-GCS) activity, and glutathione (GSH) content of spermatozoa. These results suggest that BTS supplemented with Gln can provide greater protective capacity to boar sperm against oxidative stress by enhancing GSH synthesis during liquid preservation.

      PubDate: 2018-02-26T02:57:53Z
      DOI: 10.1016/j.anireprosci.2018.02.011
  • Correlation between ubiquitination and defects of bull spermatozoa and
           removal of defective spermatozoa using anti-ubiquitin antibody-coated
           magnetized beads
    • Authors: Jian Zhang; Jie Su; Shuxiang Hu; Jindun Zhang; Rui Ding; Jitong Guo; Guifang Cao; Rongfeng Li; Qing-Yuan Sun; Xihe Li
      Abstract: Publication date: Available online 12 February 2018
      Source:Animal Reproduction Science
      Author(s): Jian Zhang, Jie Su, Shuxiang Hu, Jindun Zhang, Rui Ding, Jitong Guo, Guifang Cao, Rongfeng Li, Qing-Yuan Sun, Xihe Li
      Ubiquitination is an important cellular process in spermatogenesis and involves the regulation of spermatid differentiation and spermiogenesis. In the current study, the correlation between bull sperm ubiquitination and sperm defects was analyzed, and the feasibility using anti-ubiquitin specific antibody immobilized magnetic beads to remove the spermatozoa with defects was assessed. A total of nine bulls were examined, and the amount of sperm ubiquitination ranged from 55 to 151. Correspondingly, the percentage of sperm deformity ranged from 9.3% to 28.1%. The coefficient of correlation was r = 0.92, indicating a significant correlation between the percentage of sperm deformity and the amount of ubiquitination (P < 0.05). The results from use of fluorescence staining and single-channel flow cytometry indicated there was a significant correlation between the sperm deformity and amount of ubiquitination (r = 0.86, P < 0.05). Results gained by use of the TUNEL and ubiquitination assays by double-channel flow cytometry indicated that the proportion of genetically defective spermatozoa with ubiquitination in Q3 and Q2 quartiles was markedly greater than that of spermatozoa with ubiquitination in Q1 and Q4 quartiles (82.1% compared with 17.9%). All these results confirmed that sperm ubiquitination is associated with genetic DNA defects (P < 0.01). Furthermore, nine semen samples with sperm motility of less than 50% (minimal motility), 50% to 70% (moderate motility) and greater than 70% (greatest motility) were selected for sorting defective spermatozoa using anti-ubiquitin specific antibody-coated magnetic beads. Strikingly, the percentage of sperm deformity significantly decreased from 18.8%, 19.0% and 17.1% to 11.7%, 11.0% and 11.0%, respectively (P < 0.05), suggesting that this method might be a feasible technology to improve the productivity via removal of the defective spermatozoa from bull semen.

      PubDate: 2018-02-26T02:57:53Z
      DOI: 10.1016/j.anireprosci.2018.01.018
  • Colloid centrifugation of fresh semen improves post-thaw quality of
           cryopreserved dromedary camel spermatozoa
    • Authors: Clara Malo; Elizabeth G. Crichton; Jane M. Morrell; Budhan S. Pukazhenthi; Anders Johannisson; Rebecca Splan; Julian A. Skidmore
      Abstract: Publication date: Available online 12 February 2018
      Source:Animal Reproduction Science
      Author(s): Clara Malo, Elizabeth G. Crichton, Jane M. Morrell, Budhan S. Pukazhenthi, Anders Johannisson, Rebecca Splan, Julian A. Skidmore
      Colloids have been successfully used in a number of species to improve sperm populations for IVF and for cryopreservation The usefulness of Single Layer Centrifugation (SLC) for freezing dromedary camel spermatozoa in two different extenders was evaluated by examining the motility, viability, acrosome status, DNA integrity, and ability of cryopreserved sperm to penetrate oocytes in vitro in a heterologus IVF system. Two ejaculates from each of five males were divided into four aliquots: two were processed by SLC (selected) while two were centrifuged without colloid (control). Pellets were cryopreserved in Green Buffer or INRA-96® containing 3% glycerol and evaluated at 0 and 1 h post thawed. The SLC improved post-thaw total and progressive motility at 0 (both P < 0.0001) and 1 (P < 0.001; P < 0.01, respectively) h, and STR (both P < 0.05) and BCF (both P < 0.001) at 0 h. Sperm viability and acrosome integrity (both P < 0.001) were improved at both time points. Sperm frozen in Green Buffer had greater total and progressive motilities at 0 (both P < 0.001) and 1 (both P < 0.001) h than INRA-96® samples. Spermatozoa in Green Buffer also had a greater VAP, VCL and VSL at 0 h and improved viability and acrosome integrity at 0 h (P < 0.05; P = 0.001, respectively) and 1 h (P < 0.05; P < 0.001, respectively). Viability of SLC spermatozoa was improved in Green Buffer at 1 h (P < 0.05). Oocyte penetration (P < 0.05) and pronuclear formation (P < 0.01) were greater with SLC-selected spermatozoa than non-selected spermatozoa, regardless of extender. No difference was observed between treatments or extenders in the mean number of spermatozoa per oocyte penetrated. The SLC spermatozoa had less (P < 0.01) DNA fragmentation compared to controls. The DNA fragmentation was moderately and negatively correlated with penetration (r = -0.4162; P = 0.02) and pronuclear formation (r = -0.3390; P < 0.01). In conclusion, colloid centrifugation of spermatozoa and cryopreservation in Green Buffer improves post thaw motility variables and IVF performance of dromedary camel spermatozoa.

      PubDate: 2018-02-26T02:57:53Z
      DOI: 10.1016/j.anireprosci.2018.02.005
  • Cryopreservation and storage of cat epididymal sperm using ‒75 °C
           freezer vs liquid nitrogen
    • Authors: K. Buranaamnuay
      Abstract: Publication date: Available online 12 February 2018
      Source:Animal Reproduction Science
      Author(s): K. Buranaamnuay
      The quality of cat epididymal sperm cryopreserved and stored by four methods was assessed. Epididymal sperm were suspended in Tris-glucose-citrate egg yolk extender, loaded in 0.25 mL straws and then cryopreserved. The samples in a standard protocol (LN) were cryopreserved and stored in liquid nitrogen (LN2). The sperm straws in the LN-Fr-LN group were cryopreserved in LN2 and stored in a −75 °C freezer; the straws were returned to LN2 prior to thawing. The loaded straws in the Fr group were transferred directly from 4 °C to the freezer and maintained in the freezer until thawing. The Fr-LN samples were cryopreserved and stored in the freezer and were introduced into LN2 before thawing. The sperm thawing was conducted on days 30, 60, 90 and 120 of cryopreservation. The sperm motility, viability, membrane integrity and acrosome integrity were evaluated at 15 and 180 min after thawing. The quality of post-thaw sperm in all three modified protocols was comparable (P > 0.05) and did not differ from that in the standard protocol except the membrane integrity of the 60 days stored samples evaluated at 15 min after thawing, which was significantly higher for the LN-Fr-LN than the Fr-LN groups (P = 0.04). The length of cryopreservation time had no effect (P > 0.05) on the sperm parameters assessed at 15 min after thawing. The sperm motility was significantly greater (P = 0.01 to P = 0.02) for the 15 min than the 180 min incubation. In conclusion, cat epididymal sperm could alternatively be cryopreserved and/or stored by using the −75 °C freezer for 120 days. To use, the cryopreserved sperm in the freezer could be thawed immediately or after being transferred to LN2. This was useful for the application of the −75 °C cryopreserved sperm in remote areas.

      PubDate: 2018-02-26T02:57:53Z
      DOI: 10.1016/j.anireprosci.2018.02.008
  • Identification of genomic variants causing sperm abnormalities and reduced
           male fertility
    • Authors: Jeremy F. Taylor; Robert D. Schnabel; Peter Sutovsky
      Abstract: Publication date: Available online 10 February 2018
      Source:Animal Reproduction Science
      Author(s): Jeremy F. Taylor, Robert D. Schnabel, Peter Sutovsky
      Whole genome sequencing has identified millions of bovine genetic variants; however, there is currently little understanding about which variants affect male fertility. It is imperative that we begin to link detrimental genetic variants to sperm phenotypes via the analysis of semen samples and measurement of fertility for bulls with alternate genotypes. Artificial insemination (AI) bulls provide a useful model system because of extensive fertility records, measured as sire conception rates (SCR). Genetic variants with moderate to large effects on fertility can be identified by sequencing the genomes of fertile and subfertile or infertile sires identified with high or low SCR as adult AI bulls or yearling bulls that failed Breeding Soundness Evaluation. Variants enriched in frequency in the sequences of subfertile/infertile bulls, particularly those likely to result in the loss of protein function or predicted to be severely deleterious to genes involved in sperm protein structure and function, semen quality or sperm morphology can be designed onto genotyping assays for validation of their effects on fertility. High throughput conventional and image-based flow cytometry, proteomics and cell imaging can be used to establish the functional effects of variants on sperm phenotypes. Integrating the genetic, fertility and sperm phenotype data will accelerate biomarker discovery and validation, improve routine semen testing in bull studs and identify new targets for cost-efficient AI dose optimization approaches such as semen nanopurification. This will maximize semen output from genetically superior sires and will increase the fertility of cattle. Better understanding of the relationships between male genotype and sperm phenotype may also yield new diagnostic tools and treatments for human male and idiopathic infertility.

      PubDate: 2018-02-26T02:57:53Z
      DOI: 10.1016/j.anireprosci.2018.02.007
  • Impact of light irradiation on preservation and function of mammalian
    • Authors: Marc Yeste; Miriam Castillo-Martín; Sergi Bonet; Joan E. Rodríguez-Gil
      Abstract: Publication date: Available online 9 February 2018
      Source:Animal Reproduction Science
      Author(s): Marc Yeste, Miriam Castillo-Martín, Sergi Bonet, Joan E. Rodríguez-Gil
      Light irradiation has been demonstrated to exert positive effects on gametes, and particularly on sperm. In effect, a high number of studies conducted in several species, including humans, mice, pigs, cattle and sheep, and using different light sources (such as lasers and light-emitting diodes) have demonstrated that photo-stimulation increases sperm motility. In addition, other works have shown that sperm fertilizing ability both in vitro and in vivo can be increased following light irradiation; there are also some evidences pointing out to an extend of lifespan of preserved semen. Notwithstanding, no study has reported a detrimental effect of visible light on DNA integrity. The mechanisms through which light exerts its effects are not completely elucidated, but mounting evidence gives cell photosensitizers, especially those present in the mitochondria, a vital role. Stimulating these molecules turns into an increase in the production of ATP and Ca2+ influx, which contributes to explain the effects of light upon spermatozoa. Additionally, the presence of opsins in spermatozoa as well as the potential influence of light on the conformation of other proteins may also be involved in the sperm response to light. However, there are still a significant number of points that need to be addressed and their elucidation may contribute to increase the utilization of light irradiation for sperm preservation and ART.

      PubDate: 2018-02-26T02:57:53Z
      DOI: 10.1016/j.anireprosci.2018.02.004
  • Factors affecting differences between birth weight of littermates (BWTD)
           and the effects of BWTD on lamb performance
    • Authors: Jennifer L. Juengel; George H. Davis; Roger Wheeler; Ken G. Dodds; Peter D. Johnstone
      Abstract: Publication date: Available online 7 February 2018
      Source:Animal Reproduction Science
      Author(s): Jennifer L. Juengel, George H. Davis, Roger Wheeler, Ken G. Dodds, Peter D. Johnstone
      The objectives of this study were to determine factors affecting lamb birth weight (BWT) and differences between BWT of the largest and smallest littermates (BWTD) and to assess the relative importance of BWT and BWTD on lamb survival and growth to weaning. Records from twin (n = 5369) or triplet (n = 1664) litters born on pasture were utilised. Breeds included Coopworth, Romney, Perendale, composite and Texel. Ewe and sire breed, age and weight of the dam, sex, and number of lambs born (NLB) affected BWT. Neither embryo loss nor ovulation pattern (i.e. all ova released from one ovary or some from each) were associated with BWT. Dam weight and NLB affected BWTD but dam age, sire or dam breed did not. Loss of an embryo reduced BWTD, and BWTD was lower when the ewe ovulated from both ovaries versus only one ovary for twins, but not triplets. Whereas BWT was moderately heritable (h2 = 0.20), BWTD was not heritable (h2 = 0.003). Lamb survival was positively associated with BWT. Lambs with BWTD > 1.3 kg were less likely to survive (73.3%) than those from litters of smaller BWTD (range 82.8–85.7% survival). Growth rate of twin and triplet lambs was positively associated with BWT. Surprisingly, lambs from litters with moderately high BWTD had slightly greater (approximately 3%) growth rate than those from lambs of low to intermediate BWTD. Thus, while large BWTD were negatively associated with survival, BWT itself, which was moderately heritable, appeared to be a stronger driver of lamb survival and average daily gain.

      PubDate: 2018-02-26T02:57:53Z
      DOI: 10.1016/j.anireprosci.2018.02.002
  • Selected serum acute-phase proteins in peripartum sows and evaluation of
           their diagnostic usefulness
    • Authors: Karol Wierzchosławski; Krzysztof Kwit; Zygmunt Pejsak; Małgorzata Pomorska-Mól
      Abstract: Publication date: Available online 7 February 2018
      Source:Animal Reproduction Science
      Author(s): Karol Wierzchosławski, Krzysztof Kwit, Zygmunt Pejsak, Małgorzata Pomorska-Mól
      Lactation impairment in sows is a frequent and significant clinical problem. Due to a complex aetiopathogenesis, early diagnosis of postpartum dysgalactia syndrome (PDS) is difficult and so far has usually been based on physical examination performed in the first days after farrowing. To date no data have been provided on the diagnostic usefulness of acute phase proteins (APP) in early diagnosis of peripartum disorders, including lactation disorders in sows. This study aimed at measuring the serum concentration of selected APP (C-reactive protein (CRP), haptoglobin (Hp), serum amyloid A (SAA) and pig major acute phase protein (Pig-MAP)) in sows with physiological and pathological course of the peripartum period and at evaluating the possibility of utilising the studied markers in the early diagnosis of lactation disorders. Also, the correlation between the studied APP serum concentration and production parameters was assessed. To the best of the authors’ knowledge, the present study is the first such performed on sows. The experiment was conducted on 139 sows divided into three experimental groups based on the course of peripartum period: HEALTHY (n = 58) – clinically healthy sows, PDS (n = 45) – sows with milk production disorders, and OTHERS (n = 36) – sows which had experienced difficult parturitions, inflammations not connected with mammary glands (abscesses, hooves infections), or lameness. Thirteen serum samples from each sow were analysed, samples being taken on days -28 (-30 to -25), -14 (-16 to -11), -7 (-8 to -6), -5, -3, -1, 0 (parturition day), +1, +3, +5, +7, +14 and +28 (prior to or post farrowing). In order to measure the level of serum APP, commercial, quantitative ELISA tests were used. The results of the study indicate that the diagnosis made on the basis of the assessment of SAA levels on day 7 before the farrowing was not statistically different from the diagnosis made on the basis of the physical examination in the first days after the farrowing, that is the so-called “gold standard”. The achieved results indicate that SAA may be a useful early marker of lactation impairments in sows, which allows detection of which sows are susceptible to lactation disorders with high probability even as early as one week before parturition.

      PubDate: 2018-02-26T02:57:53Z
      DOI: 10.1016/j.anireprosci.2018.02.003
  • Determinants of gestation length in Thoroughbred mares on German stud
    • Authors: Mareike Ewert; Imke Lüders; Jozsef Böröcz; Hubert Uphaus; Ottmar Distl; Harald Sieme
      Abstract: Publication date: Available online 7 February 2018
      Source:Animal Reproduction Science
      Author(s): Mareike Ewert, Imke Lüders, Jozsef Böröcz, Hubert Uphaus, Ottmar Distl, Harald Sieme
      The aim of the present study was to analyze the effects of stallion and mare, their ages, and maternal lineage on the gestation length (GL) in Thoroughbreds. In addition, additive genetic effects of the dam, stallion and fetus were analyzed. Data were taken from 1993 through 2009, and included 16,226 pregnancies from 5959 Th oroughbred mares mated with 290 different stallions. All analyses were performed using linear mixed models. The GL ranged from 306 to 390 days, with a mean length of 347.0 ± 14.4 days. Mating of mares with stallions aged 17 years and older resulted in a significantly longer GL compared to younger stallions. Furthermore, the GL significantly increased with the increasing age of the mares, and the GL was longer with male foals. The month and year of breeding, as well as the mare´s breeding history (parity and reproductive status) also affected GL. The mare and stallion themselves explained 18% and 4% of the variance in GL. Coefficients of inbreeding of mares and foals had no significant effect on GL. The heritability for the GL was 0.17 for the dam and 0.006 for the fetus, whereas an additive genetic paternal effect was not estimable. The relative proportions among the additive genetic and permanent environmental contributions of the dam were 76.5% and 23.5%. A maternal lineage effect was not obvious.

      PubDate: 2018-02-26T02:57:53Z
      DOI: 10.1016/j.anireprosci.2018.02.001
  • Seminal plasma differentially alters the resistance of dog, ram and boar
           spermatozoa to hypotonic stress
    • Authors: Guillaume Tsikis; Karine Reynaud; Stéphane Ferchaud; Xavier Druart
      Abstract: Publication date: Available online 2 February 2018
      Source:Animal Reproduction Science
      Author(s): Guillaume Tsikis, Karine Reynaud, Stéphane Ferchaud, Xavier Druart
      During ejaculation and the deposition in the female genital tract, spermatozoa undergo hypo-osmotic stress and need to withstand it for optimal fertility. Resistance to hypo-osmotic stress may be affected by the interaction of the spermatozoa with seminal fluid components. The hypo-osmotic resistance of epididymal and ejaculated spermatozoa from dogs, rams and boars was assessed by flow cytometric measurement of sperm viability after incubation in NaCl solutions with osmolalities ranging from 0 to 300 mmol/kg. The hypotonic resistance of epididymal spermatozoa was greater than those of ejaculated spermatozoa in all three species. Among species comparison revealed that ejaculated spermatozoa from dogs were much more resistant than those from rams and boars as 80.4 ± 5.3%, 56.7 ± 4.7 and 9.6 ± 3.6% of live spermatozoa were observed following exposure to an osmolality of 90 mmol/kg in dogs, rams and boars respectively. This can be explained by the fact that dog, ram and boar differ markedly in composition of the seminal plasma owing to the presence (ram, boar) or absence (dog) of seminal vesicles. Hypotonic resistance of epididymal and ejaculated dog spermatozoa was similar whereas ram and boar spermatozoa showed a marked drop in resistance after ejaculation. The in vitro incubation of boar epididymal spermatozoa with raw seminal plasma or the seminal plasma protein fraction induced a similar loss of resistance, suggesting that seminal proteins are involved in the lack of resistance to hypotonic stress of boar ejaculated spermatozoa.

      PubDate: 2018-02-04T19:48:52Z
      DOI: 10.1016/j.anireprosci.2018.01.012
  • Effects of parity on productive, reproductive, metabolic and hormonal
           responses of Holstein cows
    • Authors: Jéssica Tatiana Morales Piñeyrúa; Santiago Rafael Fariña; Alejandro Mendoza
      Abstract: Publication date: Available online 2 February 2018
      Source:Animal Reproduction Science
      Author(s): Jéssica Tatiana Morales Piñeyrúa, Santiago Rafael Fariña, Alejandro Mendoza
      The objective of this study was to determine the effects that parity may have on production, reproduction and the metabolic status of Holstein cows managed in a production system based on total mixed ration and pasture. Primiparous (n= 22) and multiparous (n= 24) cows from a dairy farm research station in Uruguay were used in a completely randomized design. Body weight (BW), body condition score (BCS) and backfat thickness (BFT) were recorded weekly from −30 to 70 days postpartum. Milk production was measured daily, and milk composition was determined weekly. Resumption of postpartum ovarian activity and progesterone profiles were measured three times a week based on milk progesterone. Blood was collected to determine the levels of glucose, insulin-like growth factor I (IGF-1), insulin, non-esterified fatty acids (NEFA), β-hydroxybutyrate (BHB), albumin, total protein and cholesterol. Milk production and components were lower for primiparous cows (p< 0.01) than multiparous cows. Body weights were also lower in primiparous cows than in multiparous cows (p< 0.05); however, BCS and BFT were greater (p< 0.01) in primiparous cows than in multiparous cows. Primiparous cows had greater levels of glucose, insulin and IGF-1 and lower concentrations of NEFA and BHB (p<0.01) than multiparous cows. The intervals from calving to first ovulation were not affected by parity; however, primiparous cows showed less abnormal cycles (27.2%) than multiparous cows (50.0%) (p< 0.01). The present study found that a feeding system based on TMR and pasture was sufficient to produce over 25 L of milk per day without extending their calving to first ovulation interval in primiparous and multiparous cows. However, multiparous cows showed a greater imbalance in metabolic and hormonal profiles than primiparous cows, causing abnormal ovarian activity.

      PubDate: 2018-02-04T19:48:52Z
      DOI: 10.1016/j.anireprosci.2018.01.017
  • One year daily changes in fecal sexual steroids of two captive female
           cheetahs (acinonyx jubatus) in italy
    • Authors: Valentina Vernocchi; Maria Giorgia Morselli; Massimo Faustini; Gianfranco Gabai; Laura Da Dalt; Gaia Cecilia Luvoni
      Abstract: Publication date: Available online 2 February 2018
      Source:Animal Reproduction Science
      Author(s): Valentina Vernocchi, Maria Giorgia Morselli, Massimo Faustini, Gianfranco Gabai, Laura Da Dalt, Gaia Cecilia Luvoni
      The present study evaluated changes of fecal sexual steroids in two female cheetahs (Geijsha and Duchessa) in Northern Italy throughout one year. Wet feces were collected daily from two sibling animals of the same age, housed with conspecific males and managed in the same conditions, and estrogens and progestogens concentrations were analyzed by radioimmunoassay (RIA). Evidence of ovarian activity based on regular fluctuation in estrogen excretion was demonstrated in both females. None of the animals was continuously cycling, as follicular activity was interrupted by anestrous periods, during the spring and early winter. No significant increases of progestogens were recorded after the estrogen peaks, indicating that induced or spontaneous ovulations did not occur during the observation period. The wavelet decomposition evidenced the temporal pattern of ovarian activity in the two females, underlying throughout the year a more pronounced rhythmical ovarian estrogenic activity in Geijsha than in Duchessa. However, this statistical approach had a smoothing effect in depicting the hormonal patterns and the number of follicular phases might be lower than that revealed by the iterative method. In this study, RIA on wet feces performed very well to determine sexual steroid concentrations, and an ovarian activity interrupted by anestrous periods along the year in captive cheetahs co-housed in a small group was demonstrated. More information on estrous behavior of captive cheetahs were obtained in this study, but the effects of husbandry and management conditions on natural reproductive physiology of this species remain to elucidate.

      PubDate: 2018-02-04T19:48:52Z
      DOI: 10.1016/j.anireprosci.2018.01.016
  • Anti-Müllerian Hormone (AMH), antral follicle count (AFC), external
           morphometrics and fertility in Tabapuã cows
    • Authors: Renata Maculan; Tássia Louregiani Carvalho Pinto; Gabriel Miranda Moreira; Gisvani Lopes de Vasconcelos; Jesus Afonso Sanches; Ricardo Garcia Rosa; Rafael Ribeiro Bonfim; Tarcisio de Moraes Gonçalves; José Camisão de Souza
      Abstract: Publication date: Available online 24 December 2017
      Source:Animal Reproduction Science
      Author(s): Renata Maculan, Tássia Louregiani Carvalho Pinto, Gabriel Miranda Moreira, Gisvani Lopes de Vasconcelos, Jesus Afonso Sanches, Ricardo Garcia Rosa, Rafael Ribeiro Bonfim, Tarcisio de Moraes Gonçalves, José Camisão de Souza
      The intense selection of characteristics related to animal performance may affect the reproductive efficiency of beef cows negatively. Selection for fertility along with production traits is, therefore, readily justifiable. The objective of the present study was to evaluate possible relationships among AFC, serum AMH concentrations and external morphometrics with reproductive efficiency in Tabapuã (a Brazilian Bos indicus beef breed) females. Antral follicle counts and external morphometrics were measured in nulliparous (n = 162), primiparous (n = 80) and multiparous (n = 351) cows, from four farms. Age at first parturition (AFP), parturition interval (PI), maternal ability (MA), precocity, rusticity and survival (PRS) and overall reproductive efficiency (RE) were evaluated according to the Brazilian Association of Zebu Breeders (ABCZ) data bank indexes. A single blood sample per animal was collected at random stages of the reproductive cycle from a subset of animals (nulliparous = 84, primiparous and multiparous = 136) to determine serum AMH concentrations. The AFC classes were defined as lesser (<28), intermediate (28–50) and greater (>50). Correlations between all variables and the effects of parity and AFC on reproductive traits, AMH and external genitalia size were analyzed by the PROCORR and by the PROCGENMOD procedures of SAS® (SAS, Cary, NC, USA), respectively. Antral follicle count did not differ (P = 0.71) among nulliparous (38.6 ± 23.96) primiparous (47.54 ± 26.16) and multiparous (41.08 ± 25) cows and was negatively correlated with pregnancy interval (PI), such that, as PI decreased (r = −0.28; P < 0.005), AFC increased. Vulva width was not affected (P = 0.08) by parity and was larger (P < 0.05) for females in the greater AFC class (8.81 ± 0.12 cm) compared with the intermediate (8.42 ± 0.11 cm) and lesser (8.38 ± 0.13 cm) classes. As vulva width increased, parturition interval decreased (r = −0.15; P < 0.005) and overall reproductive efficiency increased (r = 0.17, p < 0.005). Thoracic depth was associated with greater AFCs (r = 0.10; P < 0.001). Anti-Müllerian hormone concentrations were greater (P < 0.05) for animals in the greater AFC class (1.15 ± 0.09 ng/mL) compared with the lesser (0.44 ± 0.02 ng/mL) and intermediate (0.73 ± 0.05 ng/mL) classes. Vulva width, AFC, external morphometrics and AMH concentrations were moderately associated, considering the low heritability of fertility traits, and should be studied further to be considered in the selection for fertility in Bos indicus cattle.

      PubDate: 2017-12-27T09:20:57Z
      DOI: 10.1016/j.anireprosci.2017.12.011
    • Authors: Anne Kemmer Souz; Luiz Guilherme Corsi Trautwein; Cristiane Sella Paranzini; Felipe Montanhero Perencin; Guilherme Schiess Cardoso; Maria Isabel Mello Martins
      Abstract: Publication date: Available online 22 December 2017
      Source:Animal Reproduction Science
      Author(s): Anne Kemmer Souz, Luiz Guilherme Corsi Trautwein, Cristiane Sella Paranzini, Felipe Montanhero Perencin, Guilherme Schiess Cardoso, Maria Isabel Mello Martins
      The objective of this study was to identify and compare domestic feline sperm subpopulations, chilled at −1 °C for 24 and 48 hours, as well as to analyze the sperm frequency in different subpopulations. Ten adult cats were used. Sperm collection was performed using electroejaculation (EEJ). Spermatic kinetics were evaluated using a computerized system at three moments: fresh, 24 and 48 hours after refrigeration. The ejaculates were divided into a group refrigerated at −1 °C (n = 5,) and a group refrigerated at 4 °C (n = 5),. A total of 1560 spermatozoa were analyzed individually, and the sperm subpopulations were identified using multivariate statistics. Three spermatic subpopulations were defined using prior analysis of the hierarchical dendrogram. A principal components analysis (PCA) identified the existence of three groups with higher iterations at the three moments: PC1 (VAP, VCL, VSL, ALH, SVI), PC2 (STR, LIN, WOB and SMI) and PC3 (BCF). Subpopulation 1, after 48 hours of refrigeration at −1 °C, and subpopulation 3, after 24 hours of refrigeration at 4 °C, maintained their sperm quality, which allowed us to characterize the groups of spermatozoa that were resistant to cryopreservation. The present study identified three well defined ejaculate spermatozoa subpopulations, with proportional distributions between the groups and two refrigeration resistant subpopulations.

      PubDate: 2017-12-27T09:20:57Z
      DOI: 10.1016/j.anireprosci.2017.12.015
  • Factors affecting staining to discriminate between bull sperm with greater
           and lesser mitochondrial membrane potential
    • Authors: Alessia Gloria; Laura Wegher; Augusto Carluccio; Claudio Valorz; Domenico Robbe; Alberto Contri
      Abstract: Publication date: Available online 21 December 2017
      Source:Animal Reproduction Science
      Author(s): Alessia Gloria, Laura Wegher, Augusto Carluccio, Claudio Valorz, Domenico Robbe, Alberto Contri
      The sperm mitochondrial membrane potential (MMP) is usually evaluated using the JC-1 dye. This study aimed to verify the effect of incubation temperature (25 °C or 38 °C), incubation time (10, 30, and 45 min), JC-1 stain concentration (0.2 μM, 2 μM, 8 μM, 12 μM), and the presence of glycerol (6.6% compared with 0%), on the capacity of the stain to discriminate between sperm with high mitochondrial membrane potential (hMMP) and low mitochondrial membrane potential (lMMP) in fresh and frozen bull sample by both flow cytometry and epifluorescence microscopy. The temperature (38 °C for 10 min) and the dye concentration (8 μM and 12 μM) resulted in a greater proportion of hMMP (P < .05). The incubation for 45 min at 38 °C resulted in a significant reduction of hMMP in samples stained with JC-1 dye at 8 μM and 12 μM (P < .01). A longer incubation time (45 min) and greater dye concentration (8 μM and 12 μM) resulted in an increased proportion of hMMP sperm in cryopreserved samples. Fresh sperm incubated with glycerol had a hMMP (P < .05). Data for the present study indicate that the optimal incubation temperature was 38 °C, with an incubation time differing between fresh (10–30 min) and cryopreserved sperm (at least 45 min). Furthermore, the JC-1 dye concentration used that could reliably detect the proportion of hMMP sperm was 2 μM in fresh samples, and at least 8 μM in cryopreserved sperm.

      PubDate: 2017-12-27T09:20:57Z
      DOI: 10.1016/j.anireprosci.2017.12.007
  • Proteomic analyses of ram (Ovis aries) testis during different
           developmental stages
    • Authors: Zengkui Lu; Youji Ma; Quanwei Zhang; Xingxu Zhao; Yong Zhang; Liping Zhang
      Abstract: Publication date: Available online 21 December 2017
      Source:Animal Reproduction Science
      Author(s): Zengkui Lu, Youji Ma, Quanwei Zhang, Xingxu Zhao, Yong Zhang, Liping Zhang
      Male reproductive capacity is essential for animal breeding and reproduction. In males, the testes produce sperm and secrete androgen, processes which require precise regulation by multiple proteins. The composition of proteins in the ram testes has not yet been studied systematically, thus, the application of proteomics to explore differential protein regulation during ram testes development is of great significance. In the present study, ram testes were studied at five different developmental phases to assess postnatal differences in protein regulation. Two dimensional electrophoresis (2-DE) was used to separate ram testes proteins at each developmental phase, yielding 45 different proteins, 37 of which were identified by Matrix Assisted Laser Desorption Ionization-Time of Flight-Time of Flight-Mass Spectrometry (MALDI-TOF/TOF-MS). Gene Ontology (GO) was used to specifically annotate the biological process, cellular composition, and molecular function of each identified protein. Most of the identified proteins were involved in structural formation, development, reproduction, and apoptosis of the testicular spermatogenic tissue and spermatozoa. Quantitative real time PCR (qRT-PCR), western blot and immunohistochemical methods were used to verify the proteins, and the results were consistent with that of 2-DE. The proteins that were different in abundance that were identified in this study can be used as biomarkers in future studies of ram reproduction.

      PubDate: 2017-12-27T09:20:57Z
      DOI: 10.1016/j.anireprosci.2017.12.012
  • Incubation of spermatozoa with Anandamide prior to cryopreservation
           reduces cryocapacitation and improves post-thaw sperm quality in the water
           buffalo (Bubalus bubalis)
    • Authors: Puneeth Kumar; Tushar Kumar Mohanty; Arumugam Kumaresan; Pradeep Nag; Kaustubh Kishor Saraf; Vimlesh Kumar; Sreela Lathika; Samiksha Nayak; Mukesh Bhakat
      Abstract: Publication date: Available online 21 December 2017
      Source:Animal Reproduction Science
      Author(s): Puneeth Kumar, Tushar Kumar Mohanty, Arumugam Kumaresan, Pradeep Nag, Kaustubh Kishor Saraf, Vimlesh Kumar, Sreela Lathika, Samiksha Nayak, Mukesh Bhakat
      Anandamide (AEA), an endocannabinoid, has been shown to reduce capacitation and acrosomal exocytosis in human spermatozoa. Because buffalo spermatozoa are highly susceptible to cryopreservation induced damage, AEA was assessed as to whether it could protect spermatozoa from cryo-damage. Six ejaculates from six Murrah buffalo bulls (total 36 ejaculates) were utilized for the study. Each ejaculate was divided into four aliquots; spermatozoa in Aliquot 1 were extended in Tris-Citrate-Egg Yolk and frozen as per the standard protocol. Spermatozoa in Aliquots 2, 3 and 4 were incubated with AEA at 1 nM, 1 μM and 10 μM, respectively in Tris-Citrate extender for 15 min at 37 °C before cryopreservation. Cryopreserved spermatozoa were thawed at 37 °C for 30 s before assessment of sperm motility, membrane integrity, capacitation, acrosome reaction, mitochondrial membrane potential (MMP) and lipid peroxidation status. The proportion of motile and membrane intact spermatozoa were greater (P < 0.05) with use of 1 μM AEA incorporated group compared with other groups. The proportion of un-capacitated and acrosome intact spermatozoa was greater (P < 0.05) with use of 1 or 10 μM of AEA compared with the other groups. When compared to the control group, use of 1 μM AEA resulted in a greater proportion of spermatozoa with high MMP (P < 0.05). There was no significant difference in the lipid peroxidation status of spermatozoa among any of the four groups. It was inferred that the protective role of AEA during cryopreservation of buffalo spermatozoa was dose dependent and incubation of spermatozoa with AEA at 1 μM concentration prior to cryopreservation reduced cryo-capacitation and improved post-thaw sperm quality in buffalo.

      PubDate: 2017-12-27T09:20:57Z
      DOI: 10.1016/j.anireprosci.2017.12.010
  • The genetic polymorphisms of TGFβ superfamily genes are associated with
           litter size in a Chinese indigenous sheep breed (Hu sheep)
    • Authors: Weimin Wang; Yongfu La; Xiang Zhou; Xiaoxue Zhang; Fadi Li; Bang Liu
      Abstract: Publication date: Available online 16 December 2017
      Source:Animal Reproduction Science
      Author(s): Weimin Wang, Yongfu La, Xiang Zhou, Xiaoxue Zhang, Fadi Li, Bang Liu
      Litter size, which is a complex economic trait in sheep, is affected by polygenes. Hu sheep are a prolific native breed in China and also an ideal resource for studying the genetic mechanism that controls litter size in sheep. In this study, we investigated the genetic effects of candidate genes on litter size in a large experimental population (n = 2021) of Hu sheep. A total of 23 single nucleotide polymorphisms (SNPs) including six reported major mutations (FecB, FecXI , FecXB , FecXH , FecGI , and FecGH ) and 17 novel SNPs in 10 candidate genes involved in reproduction were genotyped using KASPar technology. Genotyping showed that Hu sheep carry the FecB mutation, but not the FecXI , FecXB , FecXH , FecGH , and FecGI mutations. Among the remaining 18 SNPs, 16 tagged SNPs were selected based on the HAPLOVIEW program. Analysis of single marker association indicated a significant association between litter size in Hu sheep and three mutations in the TGFβ superfamily (FecB, GDF9 and TGFBR2 genes). Quantitative trait modes (QTMs) analysis revealed that the FecB and GDF9 mutations exerted an additive effect, while the mutation located within TGFBR2 gene was dominant. Linear regression analysis of the association between multiple markers and litter size indicated a correlation between homozygous ewes with the GG/AA/TT genotype and larger litter size than any of the other genotypes. In conclusion, the FecB, GDF9, and TGFBR2 polymorphisms were implicated as genetic markers of potential importance in marker-assisted selection (MAS) strategies to improve litter size in Hu sheep.

      PubDate: 2017-12-27T09:20:57Z
      DOI: 10.1016/j.anireprosci.2017.12.003
  • A simple flow cytometry protocol to determine simultaneously live, dead
           and apoptotic stallion spermatozoa in fresh and frozen thawed samples
    • Authors: M.C. Gil; C. Ortega Ferrusola; L. Anel-López; J.M. Ortiz-Rodriguez; M. Alvarez; P. de Paz; L. Anel; F.J. Peña
      Abstract: Publication date: Available online 16 December 2017
      Source:Animal Reproduction Science
      Author(s): M.C. Gil, C. Ortega Ferrusola, L. Anel-López, J.M. Ortiz-Rodriguez, M. Alvarez, P. de Paz, L. Anel, F.J. Peña
      Spermatozoa undergo apoptotic changes during the cryopreservation process. These changes, recently termed spermptosis, resemble the cryopreservation induced delayed onset of cell death observed after thawing of somatic cells. Due to its importance in cryobiology, methods to easily identify spermptotic cells are warranted. In this study, a well-validated method for identification of spermatozoa with caspase 3 activity was compared with use of the combination of Hoechst 33342 (H-42) and ethidium homodimer (Eth-1). Live, dead and apoptotic spermatozoa assessed with each method were compared using descriptive statistics and method agreement analysis. No differences were observed in the percentages of spermatozoa in each of the categories investigated with each method. Moreover the method agreement analysis indicated there were consistent findings using both methods The combination H-42/Eth-1 can be successfully used to determine apoptosis in addition to dead and live spermatozoa. Moreover the intensity of H-42 fluorescence (bright and dim populations) allows for distinguishing of live and dead sperm cells.

      PubDate: 2017-12-27T09:20:57Z
      DOI: 10.1016/j.anireprosci.2017.12.009
  • Partial deoxygenation of extender improves sperm quality, reduces lipid
           peroxidation and reactive oxygen species during cryopreservation of
           buffalo (Bubalus bubalis) semen
    • Authors: B Balamurugan; SK Ghosh; SA Lone; JK Prasad; GK Das; R Katiyar; Abdul Rahman Mustapha; Ajay Kumar; MR Verma
      Abstract: Publication date: Available online 16 December 2017
      Source:Animal Reproduction Science
      Author(s): B Balamurugan, SK Ghosh, SA Lone, JK Prasad, GK Das, R Katiyar, Abdul Rahman Mustapha, Ajay Kumar, MR Verma
      The present study was designed to investigate the effect of partial deoxygenation of extender on sperm quality, lipid peroxidation (LPO) and reactive oxygen species (ROS) in buffalo (Bubalus bubalis) during cryopreservation of semen. Semen extender was prepared freshly and split into three sub-extenders [Extender I: control (non-deoxygenated), Extender II (partially deoxygenated by using LN2 flushing) and Extender III (partially deoxygenated mechanically by vacuum pump)]. Amounts of dissolved oxygen (DO) were determined in all the three extenders and also in post-thaw semen. Ejaculates with mass motility of  ≥3+ and individual progressive motility of 70% or greater were collected from Murrah buffalo bulls and utilized in the study. Each semen sample was divided into Groups I (diluted with Extender I), II (diluted with Extender II) and III (diluted Extender III) with a maximum of 60 × 106 sperm/mL. French mini straws (0.25 mL) were filled with the extended semen samples, sealed with polyvinyl alcohol powder, kept for 3 h at 5 °C for equilibration and then stored in an automatic programmable freezer until the temperature of straws reached −145 °C followed by plunging the straws into liquid nitrogen (–196 °C). Semen samples were evaluated at pre-freeze and post-thaw stages for various variables [sperm motility, live sperm count, acrosomal integrity, hypo-osmotic swelling (HOS) response, LPO and ROS concentrations]. The mean DO was less (P < 0.05) in Extender II as compared to I and III. The DO was less (P < 0.05) in Group II (semen extended with Extender II) as compared with III (semen extended with Extender III) and I (semen extended with Extender I). The percentages for sperm motility, viability and intact acrosomes (PIA) were greater (P < 0.05) in Groups II and III as compared to the control group at the pre-freeze stage, while at the post-thaw stage, percentages of sperm motility, viability, PIA and HOS response were greater (P < 0.05) in Group II as compared with the control group and Group III. Pre-freeze HOS response (%) was greater (P < 0.05) in Group II as compared with the control and Group III. At the pre-freeze stage, sperm LPO and ROS were less (P < 0.05) in Groups II and III as compared with the control and at post-thaw stage, spermatic LPO and ROS concentrations were less (P < 0.05) in Group II than in the control group and Group III. In conclusion, partial deoxygenation of extender improves sperm quality, reduces sperm LPO and ROS concentrations in buffalo during cryopreservation. Partial deoxygenation of the extender with LN2 flushing may be one of the ways for improving quality and fertility of frozen-thawed buffalo sperm.

      PubDate: 2017-12-27T09:20:57Z
      DOI: 10.1016/j.anireprosci.2017.12.008
  • Ultrastructure of spermatozoa in cobia, Rachycentron canadum (Linnaeus,
    • Authors: Dhanasekar Krishnamoorthy; Selvakumar Narasimman; Munuswamy Natesan
      Abstract: Publication date: Available online 11 December 2017
      Source:Animal Reproduction Science
      Author(s): Dhanasekar Krishnamoorthy, Selvakumar Narasimman, Munuswamy Natesan
      Ultrastructure and development of spermatozoa in cobia, Rachycentron canadum are described. Sections through the testis show different developmental stages viz, Spermatocytes, spermatids and sperm. Spermatozoa of R. canadum exhibit the configuration of uniflagellated, anacrosomal Type I aquasperm, typical for externally fertilizing fish. Mature spermatozoon is seen with a prominent head and long cylindrical flagellum. Ultrastructure of sperm shows invaginated ‘U’ shaped nucleus and other organelles. The mitochondrial matrix is electron-dense with irregular arrangement of the cristae. The nucleus reveals a deep invagination (nuclear fossa) in which the centriolar complex is located. The centriolar complex lies inside the nuclear fossa and is composed of a proximal and a distal centriole. The two centrioles are placed perpendicular to each other. The flagellum has a typical eukaryotic organization (microtubule doublets 9+2 pattern) and measures around 36.21±0.42μm in length. This study for the first time provides a comprehensive detail on the ultrastructure and developmental process of sperm in cobia, R. canadum.

      PubDate: 2017-12-13T05:10:33Z
      DOI: 10.1016/j.anireprosci.2017.12.005
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