- Effect of Dietary N-Carbamylglutamate Levels on Reproductive Performance
- Authors: B Zhang; LQ Che, Y Lin, Y Zhuo, ZF Fang, SY Xu, J Song, YS Wang, Y Liu, P Wang, D Wu
Pages: n/a - n/a
Abstract: The effect of dietary N-carbamylglutamate (NCG) supplementation during the entire gestation on reproductive performance of gilts was determined. At the initial day of gestation, forty-five cross-bred (Landrace × Large white) gilts were randomly assigned to five groups receiving a basal diet (control group) and basal diet supplemented with 0.05%, 0.10%, 0.15% and 0.20% NCG until parturition, respectively. At parturition, total litter size, live litter size and rate of stillbirth were not markedly affected by NCG supplementation. However, gilts in 0.05% NCG-supplemented group had more pigs born alive than gilts in control group (+1.11 pigs, p = 0.12), and live litter weight was increased (+12.13–19.17%, p
- Expression Pattern of Vascular Endothelial Growth Factor in Canine
Folliculogenesis and its Effect on the Growth and Development of Follicles
after Ovarian Organ Culture
- Authors: MA Abdel-Ghani; T Shimizu, H Suzuki
Pages: n/a - n/a
Abstract: In this study, the expressions of VEGF in dog follicles were detected by immunohistochemistry and the effects of VEGF treatment on the primordial to primary follicle transition and on subsequent follicle progression were examined using a dog ovary organ culture system. The frozen-thawed canine ovarian follicles within slices of ovarian cortical tissue were cultured for 7 and 14 days in presence or absence of VEGF. After culture, the ovaries were fixed, sectioned, stained and counted for morphologic analysis. The results showed that VEGF was expressed in the theca cells of antral follicles and in the granulosa cells nearest the oocyte in preantral follicle but not in granulosa cells of primordial and primary follicles; however, the VEGF protein was expressed in CL. After in vitro culture, VEGF caused a decrease in the number of primordial follicles and concomitant increase in the number of primary follicles that showed growth initiation and reached the secondary and preantral stages of development after 7 and 14 days. Follicular viability was also improved in the presence of VEGF after 7 and 14 days in culture. In conclusion, treatment with VEGF was found to promote the activation of primordial follicle development that could provide an alternative approach to stimulate early follicle development in dogs.
- Impact of Food Restriction on Ovarian Development, RFamide-Related
Peptide-3 and the Hypothalamic–Pituitary–Ovarian Axis in
- Authors: H Li; H Song, M Huang, H Nie, Z Wang, F Wang
Pages: n/a - n/a
Abstract: RFamide-related peptide-3 (RFRP-3), the mammalian ortholog of gonadotropin-inhibiting hormone, has been implicated as a mediator between reproduction and energy balance. This study aimed to investigate the physiological effects of RFRP-3 on the process of ovarian development in food-restricted pre-pubertal ewes. The results showed that food restriction significantly inhibited the ovarian development and follicular growth. The data of qPCR in the hypothalamic–pituitary–ovarian (HPO) axis showed that food restriction not only upregulated RFRP-3 mRNA expression but also downregulated the mRNA expression of gonadotropin-releasing-hormone receptor, follicle-stimulating hormone receptor and luteinizing hormone receptor (LHR). Immunohistochemistry of RFRP-3 in the ovaries suggested that RFRP-3 may regulate the follicular development. These results suggested that the changes of RFRP-3 in response to food restriction might influence the HPO axis and inhibit ovarian development.
- Seminal Leucocytary Profile in Beef Bulls
- Authors: AL Zart; VCL Jurgielewicz, CE Fernandes
Pages: n/a - n/a
Abstract: Despite evidences that seminal leucocytes could affect semen quality, references for the frequency and concentration of these cells in bulls are not available. The aim of this study was to determine the normal concentration of leucocytes in beef bulls and to correlate this characteristic with semen quality. First, 57 bulls from artificial insemination centres were evaluated to obtain the normal leucocytary profile values. Next, 382 bulls were subjected to breeding soundness evaluation. The average concentration of leucocytes in bovine semen was 4.73 × 106/ml. Unsatisfactory bulls showed a higher number of leucocytes/field than that in the satisfactory ones. Logistic regression analysis revealed that the unsatisfactory bulls showed 6.5-fold more chances of having higher leucocyte counts than satisfactory ones. Values of up to 1 leucocyte/field in the bull ejaculate are considered physiologically normal, whereas >5 leucocytes/field is associated with poor semen quality.
- Determination of Feeder Cell-Based Cellular Niches Supporting the
Colonization and Maintenance of Spermatogonial Stem Cells from Prepubertal
Domestic Cat Testes
- Authors: NR Han; YH Park, JI Yun, HJ Park, MH Park, MS Kim, JH Choi, E Lee, SP Gong, JM Lim, ST Lee
Pages: n/a - n/a
Abstract: Recently, isolation and in vitro culture of putative spermatogonial stem cells (SSCs) in the domestic cat have been conducted. However, the cellular niche conditions that facilitate the establishment and long-term maintenance of feline SSCs (FSSCs) have not been described. Therefore, we investigated the type of feeder cells used to stimulate colony formation and growth of FSSCs among the various factors in the FSSC niche. Spermatogonial stem cells isolated from feline testes were cultured on mitotically inactivated testicular stromal cells (TSCs) derived from cats, dogs and mice, and mouse embryonic fibroblasts (MEFs). The formation and growth of colonies derived from SSCs cultured on each type of feeder cell were identified at passage 0, and the morphology, alkaline phosphatase (AP) activity and expression of SSC-specific genes in surviving colonies were investigated at passage 4. Among these diverse feeder cells, TSCs from cat showed the greatest colony formation, growth and maintenance of FSSCs, and SSC colonies cultured by passage 4 showed a typical dome-shaped morphology, AP activity and expression of SSC-specific genes (NANOG, OCT4, SOX2 and CD9). Accordingly, these results demonstrate that feline TSCs could be used as feeder cells to support the establishment and maintenance of SSCs from domestic cats.
- Effect of Three pFSH Doses on Superovulation and Embryo Quality in Goats
During Two Breeding Seasons in North-eastern Mexico
- Authors: F Sánchez-Dávila; RA Ledezma-Torres, G Padilla-Rivas, AS Bosque-González, A González Gómez, H Bernal-Barragán
Pages: e40 - e43
Abstract: The aim of this research was to evaluate the effectiveness of three pFSH doses (80 mg; 145 mg and 215 mg) on ovarian response and on quantity and quality of transferable embryos of goats during the breeding and the non-breeding seasons. Ovary structures were exposed (laparatomy under general anaesthesia) and numbers of follicles and corpora lutea were registered. Surgical embryo flushing was conducted to count and classify embryos. There were more follicles (3.4 ± 1.1) in does administered 80 mg of pFSH (p
- Comparison of Cryoprotective Effects of Lycopene and Cysteamine in
Different Cryoprotectants on Bull Semen and Fertility Results
- Authors: PB Tuncer; S Büyükleblebici, A Eken, U Taşdemir, E Durmaz, O Büyükleblebici, E Çoşkun
Abstract: The objectives of this study were to compare glycerol and ethylene glycol at different concentrations as cryoprotectants and lycopene or cysteamine (with/without) as antioxidants in Tris extender for bull semen. Twenty-four ejaculates were obtained from three bulls. Each ejaculate was split into four equal aliquots and diluted using both of the Tris extenders with glycerol (5% or 7%) or ethylene glycol (3% or 5%). After that, each extenders were split into three equal aliquots and added using both of the cysteamine 5 mm or lycopene 500 μg/ml, and control (without additives). The addition of 7% glycerol with cysteamine, 5% ethylene glycol with cysteamine and 3% ethylene glycol with cysteamine groups gave the lowest CASA motility than the other groups. However, 7% glycerol and 7% glycerol with lycopene resulted in a better rate of CASA progressive motility compared with that of other groups. Generally, all the lycopene groups signed better protective effects on acrosome and total morphology than the other groups. Glycerol 7% and 3% ethylene glycol with lycopene groups yielded to slight higher percentages of membrane integrity assessed by HOST than that of the other groups, but 7% glycerol with cysteamine and 3% ethylene glycol with cysteamine showed the worst percentages of membrane integrity. Glycerol 7% and 5% glycerol with lycopene gave rise to a higher value of VAP, VSL and VCL compared with that of the other groups. On the contrary, adding to 5% glycerol with cysteamine showed negative effect for VAP, VSL, VCL and ALH values. All cryoprotectant groups with lycopene decreased chromatin damage than the other groups. Ethylene glycol 3% led to lower non-return rates of inseminated cows. However, this result was not considered to be statistically important.
- Impact of Probing the Reproductive Tract During Early Pregnancy on
Fertility of Beef Cows
- Authors: G Pugliesi; SC Scolari, FS Mesquita, M Maturana Filho, ER Araújo, D Cardoso, JN Sales, I Martin, M Sá Filho, CM Bertan, M Binelli
Abstract: This short communication reports the impact of endometrial biopsies, uterine flushings and follicular fluid aspiration procedures at day 6 post artificial insemination (AI) on pregnancy rates. In Experiment 1, cows were timed AI (TAI) and assigned to the following treatment groups: control (n = 37), uterine flushing (n = 35) and endometrial biopsy (n = 38). On day 30 post AI, pregnancy rates were 40.5%, 33% and 28.5%, respectively (p > 0.1). Pregnancy rate on day 60 was lower (p 0.1). In Experiment 3, cows were, 6 days after TAI, randomly assigned to the following treatments: control (n = 84) or aspiration of the largest follicle (n = 73). Pregnancy rates on day 30 post AI were 63.5% for the control group and 53% for the aspirated group (p > 0.1). In conclusion, uterine flushing and endometrial biopsy negatively affect pregnancy rates, but neither procedure can be considered to be incompatible with pregnancy maintenance. Follicular aspiration during pregnancy does not interact with pregnancy success. The amount and quality of samples obtained are compatible with the use of cellular and molecular analysis of uterine variables from cows that failed or succeeded on maintaining pregnancy.
- Does Coxiella burnetii Affect Reproduction in Cattle? A Clinical
- Authors: I Garcia-Ispierto; J Tutusaus, F López-Gatius
Pages: 529 - 535
Abstract: Q fever is a zoonosis produced by Coxiella burnetii, a bacterium that is widely distributed worldwide. Domestic ruminants are the most important source of C. burnetii for human infection. In sheep and goats, abortion is the main clinical consequence of infection, yet the symptoms described in cattle have so far been inconsistent. Q fever has been also scarcely reported in cattle, most likely because of its difficult diagnosis at the farm level and because of the many existing responsible C. burnetii strains. In this report, the effects of C. burnetii infection or Q fever disease on the reproductive behaviour of dairy cattle are reviewed, with special emphasis placed on the scarcity of data available and possible control actions discussed.
- Are Pharmacological Interventions Between Conception and Birth Effective
in Improving Reproductive Outcomes in North American Swine'
- Authors: JM Wessels; K Khalaj, RT Kridli, AK Edwards, M Bidarimath, C Tayade
First page: 536
Abstract: The objective of this review is to evaluate the effectiveness of using pharmacological compounds on reproductive outcomes, particularly litter size, in North American swine. While the opportunity to improve reproduction in North American pigs exists, numerous hurdles need to be overcome in order to achieve measureable results. In the swine industry, the majority of piglet losses are incurred during pregnancy and around farrowing. Over the last 20 years, a reduction in losses has been achieved through genetic selection and nutritional management; however, these topics are the focus of other reviews. This review will evaluate attempts to improve litter size by reducing losses at various stages of the reproductive process, from the time of conception to the time of farrowing, using pharmacological compounds. Generally, these compounds are used to either alter physiological processes related to fertilization, embryonic attachment or uterine capacity, etc., or to facilitate management aspects of the breeding females such as inducing parturition. Although some of the pharmacological agents reviewed here show some positive effects on improving reproductive parameters, the inconsistent results and associated risks usually outweigh the benefits gained. Thus, at the present time, the use of pharmacological agents to enhance reproduction in North American swine may only be recommended for herds with low fertility and presents an avenue of research that could be further explored.
- Apoptosis-Like Events and In Vitro Fertilization Capacity of Sex-Sorted
- Authors: X-M Zhao; J-J Ren, S-J Zhao, L-S Cui, H-S Hao, H-Y Wang, W-H Du, T Qin, Y Liu, D Wang, H-B Zhu
First page: 543
Abstract: This study utilized three staining assays (Annexin V, mitochondrial membrane potential (JC-1) and TUNEL) for flow cytometric analysis of apoptosis in sex-sorted sperm from four different bulls (A, B, C and D). Correlations between sperm quality and IVF efficiency were then assessed to determine which assay provided the best prediction of IVF efficiency. The results of the Annexin V assays, as well as measures of viable sperm, early apoptosis, necrotic sperm and mitochondrial membrane potential (∆ψm) showed that the sex-sorted sperm collected from bull A significantly differed from those of the other three bulls (p
- The Endometrium of Cycling Cows Contains Populations of Putative
Mesenchymal Progenitor Cells
- Authors: J Cabezas; E Lara, P Pacha, D Rojas, D Veraguas, F Saravia, L Rodríguez-Alvarez, FO Castro
Pages: 550 - 559
Abstract: Endometrial stem cells have been identified in humans, mice and pigs. This study was designed to determine whether the uterine endometrium of cycling cows contains such cells, to identify markers of stemness and ultimately to isolate putative stem/progenitor cell and evaluate their capability to differentiate into mesodermal derivatives. Uteri from healthy cows in the early (days 1–5) and late luteal phases (days 13–18) of the oestrous cycle were collected. Total RNA and proteins were isolated and searched for gene markers of embryonic (OCT4, NANOG, SOX2) and mesenchymal (CD44, STAT3, CD-117) stem cells and for protein markers (Oct4, Sox2, Cd44) in Western blots or immunostaining of paraffin-embedded tissue. Primary cell cultures were isolated; characterized in terms of morphology, colony formation and gene/protein expression; and induced osteogenic and chondrogenic differentiation. We identified expression of embryonic (OCT4 and SOX2, but not NANOG) and mesenchymal (STAT3, CD44 and c-KIT) gene markers in the endometrium of cycling cows and the encoded proteins (Oct4, Sox2 and Cd44) in both stages of the oestrous cycle. Derived cell lines displayed essentially the same gene expression pattern; however, at the protein level, Oct4 was not detected. No clear influence of the stage of the oestrous cycle was found. Cell lines from late luteal phase displayed osteogenic and chondrogenic differentiation potential upon chemical stimulation. In this research, we demonstrated the presence of mesenchymal progenitor cell populations of apparently mesenchymal origin in the endometrium of cycling cows, in both the early and late phases of the oestrous cycle. The cells isolated from the late luteal phase were more acquiescent to differentiate into mesodermal derivatives than cells in the early luteal phase. Our findings might have implications for the understanding of uterine stem cell biology in cows and other farm animal species.
- Kinematics and Subpopulations’ Structure Definition of Blue Fox
(Alopex lagopus) Sperm Motility using the ISAS® V1 CASA System
- Authors: C Soler; A García, J Contell, J Segervall, M Sancho
First page: 560
Abstract: Over recent years, technological advances have brought innovation in assisted reproduction to the agriculture. Fox species are of great economical interest in some countries, but their semen characteristics have not been studied enough. To advance the knowledge of function of fox spermatozoa, five samples were obtained by masturbation, in the breeding season. Kinetic analysis was performed using ISAS® v1 system. Usual kinematic parameters (VCL, VSL, VAP, LIN, STR, WOB, ALH and BCF) were considered. To establish the standardization for the analysis of samples, the minimum number of cells to analyse and the minimum number of fields to capture were defined. In the second step, the presence of subpopulations in blue fox semen was analysed. The minimum number of cells to test was 30, because kinematic parameters remained constant along the groups of analysis. Also, the effectiveness of ISAS® D4C20 counting chamber was studied, showing that the first five squares presented equivalent results, while in the squares six and seven, the kinematic parameters showed a reduction in all of them, but not in the concentration or motility percentage. Kinematic variables were grouped into two principal components (PC). A linear movement characterized PC1, while PC2 showed an oscillatory movement. Three subpopulations were found, varying in structure among different animals.
- The Effect of the Intracervical Application of Follicle-Stimulating
Hormone or Luteinizing Hormone on the Pattern of Expression of
Gonadotrophin Receptors in the Cervix of Non-Pregnant Ewes
- Authors: S Leethongdee; M Khalid, RJ Scaramuzzi
Pages: 568 - 575
Abstract: During the periovulatory period, the cervix relaxes in response to changes in circulating concentrations of reproductive hormones. The present study investigated the role of gonadotrophins in cervical function by examining the expression of follicle-stimulating hormone receptor (FSHR) and luteinizing hormone receptor (LHR) and their mRNAs following intracervical treatment with either FSH or LH. Eighteen ewes were assigned to four groups, and they were then treated with progestagen sponges and PMSG to synchronize their oestrous cycles. Intracervical treatments were given 24 h after sponge removal as follows: Group 1: FSH 2 mg; Group 2: LH 2 mg; Group 3: Vehicle and Group 4: Control. Cervices were collected 54 h after sponge removal and then divided into three regions. The expression of FSHR and LHR was determined by immunohistochemistry and FSHR mRNA and LH mRNA by in situ hybridization. The expression of LHR, FSHR and their respective mRNAs was compared in six tissue layers (luminal epithelium, subepithelial stroma, circular, longitudinal and transverse muscle and serosa) and in three cervical regions (vaginal, mid and uterine). The results showed that FSH increased transcription of the FSHR gene and the levels of its receptor, but only in subepithelial stroma of the cervix. FSH also increased the levels of LHR in the cervix, but only in the muscle layers. LH had no effect on the levels of FSHR despite the fact that it did increase the level of transcription of the FSHR gene and LH also increased the levels of its own receptor in the cervix, but only in the muscle layers, and this action was independent of increased levels of transcription of the LHR gene. These findings suggest multiple levels of regulation of cervical LH and FSH receptors and that the gonadotrophins may have a role in relaxation of the cervix during oestrus by regulating their own receptors.
- The Influence of Morphology, Follicle Size and Bcl-2 and Bax Transcripts
on the Developmental Competence of Bovine Oocytes
- Authors: THC De Bem; PR Adona, FF Bressan, LG Mesquita, MR Chiaratti, FV Meirelles, CLV Leal
First page: 576
Abstract: This study analysed two non-invasive oocyte selection methods in relation to in vitro embryo development capacity and expression of apoptosis-related genes. Selection was based on morphological quality of oocytes or follicle diameter. Oocytes were classified as grade I (GI ≥3 layers compact cumulus cells and homogeneous cytoplasm; grade II (GII ≤3 layers compact cells and homogeneous cytoplasm;, and grade III (GIII ≥3 layers, but cells with slight expansion and slightly granulated cytoplasm). Blastocyst development was lower for GII (28.5%) than for GIII (47.7%, p 0.05). Relative expression of Bcl-2 gene was lower in the GI (1.0, p 0.05). There was no difference (p > 0.05) between GI (1.0), GII (0.92) and GIII (0.93) regarding the Bax transcript. However, the Bax and Bcl-2 transcript ratios in GII (Bax; 0.92 and Bcl-2; 1.8) and GIII (Bax; 0.93 and Bcl-2; 2.2) were different (p 0.05). Regarding Bax and Bcl-2 expression, the oocytes were similar (p > 0.05) for 1–3 mm (Bax; 1.0 and Bcl-2; 1.0), 3–6 mm (Bax; 1.0 and Bcl-2; 0.93) and 6–9 mm (Bax; 0.92 and Bcl-2; 0.91). In conclusion, oocyte selection based on morphological appearance does not guarantee the success of embryonic development. Additionally, the absence of apoptosis is not necessarily a benefit for the development of oocytes. Bovine COCs with initial signs of atresia may be used for the in vitro production of embryos, and COCs taken from follicles>3 mm in diameter are better suited to in vitro embryo development.
- Genetic Parameters and Breeding Values for Semen Characteristics in
- Authors: D Labitzke; H Sieme, G Martinsson, O Distl
First page: 584
Abstract: The objectives of this study were to show whether semen traits of 30 Hanoverian stallions regularly used in AI may be useful for breeding purposes. Semen characteristics were studied using 15 149 ejaculates from 30 Hanoverian stallions of the State Stud Celle of Lower Saxony. Semen samples were collected between 2005 and 2009. Traits analysed were gel-free volume, sperm concentration, total and motile sperm number and progressive motility. A linear multivariate animal model was employed to estimate heritabilities and permanent environmental variances for stallions. The same model was used to predict breeding values for all traits simultaneously. Heritabilities were high for gel-free volume (h2 = 0.43) and moderate for total number of sperm (h2 = 0.29) and progressive motility (h2 = 0.20). Gel-free volume, sperm concentration and total number of sperm were genetically negatively correlated with progressive motility. The effect of the permanent environment for stallions accounted for 9–55% of the trait variance. The total variance among stallions explained 37–69% of the trait variance. The average reliabilities of the breeding values were 0.43–0.76 for the 30 Hanoverian stallions. In conclusion, the study could demonstrate large effects of stallions, routinely employed in a breeding programme, on semen characteristics analysed here. We could demonstrate that estimated breeding values (EBV) with sufficient high reliabilities can be predicted using data from these stallions and these EBV are useful in horse breeding programmes to achieve genetic improvement in semen quality.
- Induction of Ram Bone Marrow Mesenchymal Stem Cells into Germ Cell Lineage
using Transforming Growth Factor-β Superfamily Growth Factors
- Authors: M Ghasemzadeh-Hasankolaei; MA Sedighi-Gilani, MB Eslaminejad
Pages: 588 - 598
Abstract: Several studies have proposed that in vitro generation of germ cells (GCs) from stem cells can be considered a future option for infertility treatment. Mesenchymal stem cells (MSCs) have the capability to differentiate into male GCs with the use of inducers such as retinoic acid. Transforming growth factor-beta 1 (TGFb1) has been shown to play important roles in male fertility and spermatogenesis. Bone morphogenic protein 4 (BMP4) and BMP8b are also involved in the derivation of primordial GCs (PGCs) from epiblast cells. Therefore, this study aims to determine whether TGFb1, BMP4 and BMP8b can initiate transdifferentiation of MSCs into GCs in vitro and to determine the type of changes that occur in the expression of GC-specific markers. In this study, we have divided passage-3 ram bone marrow (BM)-MSCs into three main groups (BMP4, BMP8b and TGFb1) which were separately treated with 10 ng/ml TGFb1, 100 ng/ml BMP4 and 100 ng/ml BMP8b for a period of 21 days. We have evaluated the ability of these groups to differentiate into GCs by assessing expressions of GC-specific markers with reverse transcription PCR (RT-PCR), quantitative RT-PCR (qRT-PCR), immunocytochemistry, morphological changes and alkaline phosphatase (ALP) activity. Our results showed that BMP4 and BMP8b induced PGCs properties in some BM-MSCs and TGFb1 formed spermatogonial stem cells (SSCs) and spermatogonia-like cells in BM-MSCs culture. The important results of this study provide the basis for additional studies to determine the exact mechanism of GCs differentiation and possibly solve the problem of infertility.
- Improvement in In Vitro Fertilization Rate, Decrease in Reactive Oxygen
Species and Spermatozoa Death Incidence in Rams by Dietary Fish Oil
- Authors: A Matini Behzad; B Ebrahimi, AR Alizadeh, V Esmaeili, A Dalman, L Rashki, AH Shahverdi
Pages: 599 - 605
Abstract: Our aim was to evaluate the effects of fish oil feeding on sperm classical parameters, level of reactive oxygen spices (ROS), spermatozoa death incidence and in vitro fertilization (IVF) rate in rams. We randomly assigned nine rams, into two experimental groups (isoenergetic and isonitrogenous rations with constant level of vitamin E supplement): control (CTR; n = 5) and fish oil (FO; n = 4, 35 g/day/ram). Diets were fed for 70 days during the physiological breeding season. After a 21-day dietary adaptation period, semen was collected weekly from each ram by an artificial vagina. Sperm classical parameters were determined by the computer-assisted sperm analyzer system (CASA), and it was prepared for IVF process by swim-up technique. These evaluations were performed during the first and last weeks of sampling. Intracellular ROS level and spermatozoa death incidence were detected by flow cytometry on a weekly basis after adaptation. Data were analysed with SPSS 15. The volume, concentration (3.6 and 2.7 × 109/ml) and sperm progressive motility (60 and 48%) were significantly improved in the FO group compared with the CTR (p
- Coexistence of Tuberculosis and Mammary Carcinoma in a Goat
- Authors: H Quintas; N Alegria, A Mendonça, A Botelho, A Alves, I Pires
First page: 606
Abstract: Synchronic occurrence of tuberculosis mastitis and mammary cancer is rare in humans and, to the best of our knowledge, not reported in domestic animals. Here, we present a case of a female adult goat of Serrana breed with simultaneous occurrence of a granulomatous mastitis, due to Mycobacterium caprae, and a mammary carcinoma. Both pathological conditions are rare in goats and should be included in differential diagnosis of mammary lesions.
- The Effects of Isoflavones on Androgens and Glucocorticoids During Puberty
on Male Wistar Rats
- Authors: S Caceres; G Silvan, L Martinez-Fernandez, MJ Illera, P Millan, B Monsalve, L Peña, JC Illera
First page: 611
Abstract: Isoflavones are the most common phytoestrogens found in human diets. However, it is still not clear whether isoflavones have effects on the reproductive and the endocrine systems under normal dietary intake and overdose. The aim of this study was to determine how the most important isoflavones, genistein and daidzein, affect androgen and glucorticoid levels on male prepuberal rats. A hundred and seventy-five 30-day-old male Wistar rats were dosed orally by stomach tube every day for 35 days, with saline solution, low and high doses of genistein, daidzein and a mixture of both. Serum samples were analysed by an enzyme immunoassay for hormone determinations. In control group, there was a peak of testosterone (T) and dihydrotestosterone levels associated to the onset of puberty, at the third week. However, in low-dose groups, the same peak was found at the fourth week (p
- Pregnancy-Associated Glycoprotein (PAG) Profile of Holstein–Friesian
Cows as Compared to Dual-Purpose and Beef Cows
- Authors: M Shahin; M Friedrich, M Gauly, W Holtz
First page: 618
Abstract: Pregnancy-associated glycoproteins (PAGs) are produced by mono- and binucleate trophoblast cells in the ruminant placenta. PAG appears in maternal blood and, from approximately 4 weeks after fertilization onward, may serve as a reliable means of diagnosing pregnancy. A range of factors are said to affect plasma PAG concentrations, such as number and sex of foetus, mass of calf and placenta, level of milk production and genetic constitution. In this study, PAG pregnancy profiles of a dual-purpose (Simmental) and two beef breeds (Uckermark and Aubrac) are compared with the profile of the specialized dairy breed Holstein–Friesian. Holstein–Friesian cows were sampled weekly; the levels of the other breeds were presented at 3-week intervals. The overall significant breed difference (p = 0.013) was founded on deviations during the initial 3 weeks of pregnancy and from 23 weeks onward. During the period critical for the detection of pregnancy, between four and 22 weeks, agreement between PAG levels of various breeds was close (p > 0.05). No significant effect of body mass of cow or calf (relative to mass of dam) was detected. These findings imply that the PAG pregnancy test may be executed uniformly irrespective of breed or type of cow, affirming the suitability of the test as a valuable asset for the cattle industry.
- Evaluation of Oestrous Detection in Dairy Cattle Comparing an Automated
Activity Monitoring System to Visual Observation
- Authors: I Michaelis; O Burfeind, W Heuwieser
First page: 621
Abstract: The objective of the study was to evaluate an automated activity monitoring (AAM) system for oestrous detection in dairy cows. Specifically, we set out to determine the oestrous detection efficiency and accuracy of the AAM system and to compare reproductive performance of cows detected either by AAM, by visual observation (VO) or by at least 1 of the 2 methods (EOM). A total of 1004 potential cow-periods from 348 cows were analysed. Oestrous detection rates (EDR) were calculated for 21 day (VWP+21), 42 day (VWP+42), and 63 day (VWP+63) after voluntary waiting period (VWP). Oestrous detection rate did not differ between AAM (42.1%) and VO (37.3%) during VWP+21 (p > 0.05) but was significantly higher in EOM (56.3%, p
- Effect of Exposure to Heatwave During Blastocyst Formation on Reproductive
Performance of Female Rabbits
- Authors: F Marco-Jiménez; C Naturil-Alfonso, DS Peñaranda, E Jiménez-Trigos, FJ García-Diego, JS Vicente
First page: 629
Abstract: We examined the effect of female exposure to heatwave during blastocyst formation on their reproductive performance and its effect on transcriptome in blastocyst and endometrial tissue. In this study, a total of 72 rabbit does were artificially inseminated and divided into two environmental groups 2 days later: does under conventional conditions (maintained between 14–22°C, n = 29) and does heat stressed in a climatic chamber (maintained between 32–37°C, n = 43). The heat-stressed group were kept under these conditions for 3 days and returned to conventional conditions thereafter. Five days post-insemination, 48 does were slaughtered to collect blastocyst and endometrium samples. mRNA transcripts from OCT-4, VEGF, erbB3, Ifn-ɣ, HSP70 and HSP90 were analysed by qRT-PCR. At day 12 of gestation, 24 females were examined by laparoscopy to evaluate implanted embryos and at birth the total kits born and individual weights were recorded. Results revealed no gene expression changes in blastocyst and endometrial tissue under heatwave exposure. Moreover, our results demonstrated that rabbit embryos developed from 8–16 cells to blastocyst during a heatwave did not affect implantation rates, total number of kits born and foetal losses. In summary, these results demonstrate that heatwave period is not a critical point in the reproductive performance of rabbits during blastocyst formation.
- ATP Content, Oxidative Stress and Motility of Beluga (Huso huso) Semen:
Effect of Short-Term Storage
- Authors: MS Aramli
First page: 636
Abstract: An effective technique for short-term storage of semen is essential when processing multiple sperm samples and when semen must be transported from collection sites to hatcheries for the fertilization of ova or to laboratories for cryopreservation. In this experiment, beluga (Huso huso) sperm were used to evaluate the effects of short-term storage on several quality parameters (i.e. motility, adenosine triphosphate (ATP) content and oxidative stress indices). Sperm cells exhibited> 50% motility during 3 days of storage with an average total duration of sperm motility varying from 13.33 ± 5.77 to 278.33 ± 25.65 s, and no motile spermatozoa were recorded after 9 days of storage. The levels of oxidative stress indices (thiobarbituric acid reactive substances and carbonyl derivatives of proteins) and antioxidant activity (superoxide dismutase) increased significantly after 3 days of storage. The ATP content also decreased significantly after 2 days of storage. The results of this study can be used to develop effective reproduction management and cryopreservation protocols for this endangered fish.
- Measurement of C-reactive protein and Prostaglandin F2α Metabolite
Concentrations in Differentiation of Canine Pyometra and Cystic
- Authors: SO Enginler; A Ateş, B Diren Sığırcı, BH Sontaş, K Sönmez, E Karaçam, H Ekici, G Evkuran Dal, A Gürel
First page: 641
Abstract: Canine pyometra is a dioestrus period disease in which systemic inflammatory response syndrome (SIRS) is a common outcome due to the response of the body to the bacterial infection. The purpose of this study was i) to differentiate canine pyometra and cystic endometrial hyperplasia (CEH)/mucometra by measuring serum C-reactive protein (CRP) and prostaglandin F2α metabolite (PGFM) concentrations in blood and ii) to compare serum concentrations of CRP and PGFM in bitches with a pathological uterus (pyometra or CEH/mucometra) to concentrations in bitches with a healthy uterus. Mean CRP concentrations were found significantly higher (p
- Systemic Immunosuppression by Methylprednisolone and Pregnancy Rates in
Goats Undergoing the Transfer of Cloned Embryos
- Authors: C Feltrin; CA Cooper, N Mohamad-Fauzi, VHV Rodrigues, LH Aguiar, S Gaudencio-Neto, LT Martins, CEM Calderón, AS Morais, IS Carneiro, TM Almeida, ING Silva, JL Rodrigues, EA Maga, JD Murray, AB Libório, LR Bertolini, M Bertolini
First page: 648
Abstract: The presence of the zona pellucida has been perceived as a requirement for the oviductal transfer of cloned embryos at early stages of development while protecting the embryo from an immune system response. We hypothesized that steroid hormone therapy could reduce a potential cellular immune response after the transfer of zona-free cloned embryos into the oviduct of recipient female goats. In Experiment 1, seven does were used to study the systemic immunosuppressant effect of the methylprednisolone administration (for 3 days) on blood cell counts. Whole blood was collected prior to treatment with methyprednisolone and then on Days 1, 2, 3, 4, 7, 14, 21 and 28 after the first dose of methylprednisolone for the analysis of haematological parameters. Methylprednisolone treatment significantly reduced circulating white blood cells and neutrophils in comparison with pre-treatment levels, demonstrating a systemic immunosuppressant effect. In Experiment 2, a group of 58 does were used as recipient females to study the effect of administration of methylprednisolone for 3 days on the establishment of pregnancies after the transfer of zona-free cloned embryos into the oviducts. No effects on pregnancy rates on Day 30 were observed regarding the distinct treatment groups (control vs. methylprednisolone), the source of oocytes (in vivo- vs in vitro-matured) or the presence or absence of the zona pellucida in embryos. In summary, methylprednisolone was effective at inducing a systemic immunosuppressed state in goats, but the treatment prior to embryo transfer did not affect pregnancy rates. Moreover, pregnancy rates were similar between zona-free and zona-intact goat cloned embryos.
- Caspase Activation, Hydrogen Peroxide Production and Akt Dephosphorylation
Occur During Stallion Sperm Senescence
- Authors: JM Gallardo Bolaños; C Balao da Silva, P Martín Muñoz, M Plaza Dávila, J Ezquerra, IM Aparicio, JA Tapia, C Ortega Ferrusola, FJ Peña
First page: 657
Abstract: To investigate the mechanisms inducing sperm death after ejaculation, stallion ejaculates were incubated in BWW media during 6 h at 37°C. At the beginning of the incubation period and after 1, 2, 4 and 6 h sperm motility and kinematics (CASA), mitochondrial membrane potential and membrane permeability and integrity were evaluated (flow cytometry). Also, at the same time intervals, active caspase 3, hydrogen peroxide, superoxide anion (flow cytometry) and Akt phosphorylation (flow cytometry) were evaluated. Major decreases in sperm function occurred after 6 h of incubation, although after 1 h decrease in the percentages of motile and progressive motile sperm occurred. The decrease observed in sperm functionality after 6 h of incubation was accompanied by a significant increase in the production of hydrogen peroxide and the greatest increase in caspase 3 activity. Additionally, the percentage of phosphorylated Akt reached a minimum after 6 h of incubation. These results provide evidences that sperm death during in vitro incubation is largely an apoptotic phenomena, probably stimulated by endogenous production of hydrogen peroxide and the lack of prosurvival factors maintaining Akt in a phosphorylated status. Disclosing molecular mechanisms leading to sperm death may help to develop new strategies for stallion sperm conservation.
- Ghrelin Accelerates In Vitro Maturation of Bovine Oocytes
- Authors: E Dovolou; IE Messinis, E Periquesta, K Dafopoulos, A Gutierrez-Adan, GS Amiridis
First page: 665
Abstract: Ghrelin, apart from its metabolic role, is nowadays considered as a basic regulator of reproductive functions of mammals, acting at central and gonadal levels. Here, we investigated for possible direct actions of ghrelin on in vitro maturation of bovine oocytes and for its effects on blastocyst yield and quality. In experiment 1, cumulus oocyte complexes (COCs) were matured in the presence of four different concentrations of ghrelin (0, 200, 800 and 2000 pg/ml). In vitro fertilization and embryo culture were carried out in the absence of ghrelin, and blastocyst formation rates were examined on days 7, 8 and 9. In experiment 2, only the 800 pg/ml dose of ghrelin was used. Four groups of COCs were matured for 18 or 24 h (C18, Ghr18, C24 and Ghr24), and subsequently, they were examined for oocyte nuclear maturation and cumulus layer expansion; blastocysts were produced as in experiment 1. The relative mRNA abundance of various genes related to metabolism, oxidation, developmental competence and apoptosis was examined in snap-frozen cumulus cells, oocytes and day-7 blastocysts. In experiment 1, ghrelin significantly suppressed blastocyst formation rates. In experiment 2, more ghrelin-treated oocytes matured for 18 h reached MII compared with controls, while no difference was observed when maturation lasted for 24 h. At 18 and 24 h, the cumulus layer was more expanded in ghrelin-treated COCs than in the controls. The blastocyst formation rate was higher in Ghr18 (27.7 ± 2.4%) compared with Ghr24 (17.5 ± 2.4%). Differences were detected in various genes’ expression, indicating that in the presence of ghrelin, incubation of COCs for 24 h caused over-maturation (induced ageing) of oocytes, but formed blastocysts had a higher hatching rate compared with the controls. We infer that ghrelin exerts a specific and direct role on the oocyte, accelerating its maturational process.
- Husbandry Factors and the Resumption of Luteal Activity in Open and
Zero-Grazed Dairy Cows in Urban and Peri-Urban Kampala, Uganda
- Authors: BM Kanyima; R Båge, DO Owiny, T Ntallaris, J Lindahl, U Magnusson, MG Nassuna-Musoke
First page: 673
Abstract: The study investigated the influence of selected husbandry factors on interval to resumption of post-partum cyclicity among dairy cows in urban and peri-urban Kampala. A prospective study of 85 day post-partum period of 59 dairy cows in open (n = 38) and zero grazing (n = 21) systems was conducted on 24 farms. Cows of parity 1–6 were recruited starting 15–30 days post-partum. Progesterone (P4) content in milk taken at 10–12 day intervals was analysed using ELISA. The cow P4 profiles were classified into ‘normal’ ( 56 days), ‘ceased’ or ‘prolonged’ (if started
- An Immunohistochemical Observations on the Oviduct of the Goat
- Authors: E Marettová; M Maretta
First page: 679
Abstract: The oviduct has an important role in regulating transport of gametes and fertilization. The main role in these functions has a smooth muscle cells and ciliated epithelium lining the oviduct. All functions are under the influence of hormonal and nervous system. The objective of this study was immunohistochemically to examine the following structures: lining epithelium, smooth muscle cells, elastic fibres and nerve fibres. For this purpose, the following antibodies were used: cytokeratin 18, S-100 protein, acetylated α-tubulin, smooth muscle actin, desmin and elastin. Ciliary and secretory cells of the lining epithelium were positive for cytokeratin 18 and S-100 protein. Cilia and the basal body-associated structures of ciliary cells were positive to acetylated α-tubulin. Smooth muscle cells (SMC) in mucosa and of the muscular layer were positive for α-smooth muscle actin (SMA) and desmin. High density of nerve fibres positively reacted to acetylated α-tubulin and S100 protein was present in the mucosa, muscular layer and serosa. Elastic fibres positive for elastin form a dense network at the base of the mucosal folds and in the muscle layer. A dense network of these fibres is accompanying the blood vessels. It is supposed that together with smooth muscle cells they are involved in the transport of ovum and in blood flow regulation.
- Forced Collapse of the Blastocoel Cavity Improves Developmental Potential
in Cryopreserved Bovine Blastocysts by Slow-Rate Freezing and
- Authors: S-H Min; J-W Kim, Y-H Lee, S-Y Park, P-S Jeong, J-Y Yeon, H Park, K-T Chang, D-B Koo
First page: 684
Abstract: This study was conducted to evaluate the effectiveness of forced collapse of the blastocoel before slow-rate freezing and vitrification of bovine blastocysts. Cryopreservation of bovine blastocysts has been proposed as a tool to improve the feasibility of cattle production using the embryo transfer technique. However, the low efficiency of frozen–thawed embryos survival and further development is a crucial problem. In this study, bovine in vitro and in vivo blastocysts were slow-rate frozen and vitrified after forced blastocoele collapse (FBC) of the blastocyst cavity by puncturing the blastocoele with a pulled Pasteur pipet. Differences in the developmental potential of frozen–thawed blastocysts derived from FBC and non-FBC groups were found in both slow-rate freezing and vitrification. Furthermore, we found that the total cell number of blastocysts in FBC groups was increased and the index of apoptosis in FBC groups was decreased. Consistent with these results, real-time RT-PCR analysis data showed that expression of the anti-apoptotic Bcl-XL gene was significantly increased by FBC groups, whereas expression of the pro-apoptotic Bax gene was significantly decreased by FBC groups. Our results also showed that pregnancy outcomes in both slow-rate frozen and vitrified bovine in vivo blastocysts could be improved by reducing the fluid content after FBC of the blastocyst cavity. Therefore, we suggest that FBC of the blastocyst cavity with a pulled Pasteur pipet is an effective pre-treatment technique for both slow-rate freezing and vitrification of bovine blastocysts.
- Evaluation of Cheetah and Leopard Spermatozoa Developmental Capability
after Interspecific ICSI with Domestic Cat Oocytes
- Authors: LN Moro; AJ Sestelo, DF Salamone
Pages: 693 - 700
Abstract: The ICSI procedure is potentially of great value for felids, and it has not been extensively studied in these species. The objectives of this work were to determine the best conditions for ICSI in the domestic cat (DC) to generate interspecific embryos by injecting cheetah (Ch) and leopard (Leo) spermatozoa. Firstly, DC oocytes were matured with insulin–transferrin–selenium (ITS) or without it (MM) and cultured using atmospheric (21%) or low (5%) oxygen tension after ICSI. The group ITS-5%O2 showed the highest blastocyst rate (p
- Quantity of IL-2, IL-4, IL-10, INF-γ, TNF-α and KC-Like
Cytokines in Serum of Bitches With Pyometra in Different Stages of
Oestrous Cycle and Pregnancy
- Authors: GS Maciel; RR Uscategui, VT Almeida, MEF Oliveira, MAR Feliciano, WRR Vicente
Pages: 701 - 704
Abstract: The occurrence of the pyometra is most common in the first half of the dioestrus when there is decreased cellular immunity associated with increased serum concentration of progesterone in females. The aim of this study was to determine the immunological profile of bitches with pyometra, studying serum levels of IL-2, IL-4, IL-10, IFN-γ, KC-like and TNF-α and comparing them with those of healthy bitches in anoestrus, dioestrus and pregnant. Forty females were divided into four experimental groups: group 1 (G1): with pyometra (n = 10); group 2 (G2): bitches in the second week of gestation (n = 10); group 3 (G3): in anoestrus (n = 10); and group 4 (G4): in dioestrus (n = 10). The serum levels for IL-2, KC-like, INF-γ and TNF-α were similar for all experimental groups. The values obtained for IL-10 were found increased (p
- Neuroendocrine, Metabolic and Genomic Cues Signalling the Onset of Puberty
- Authors: CA Meza-Herrera; A Gonzalez-Bulnes, RT Kridli, M Mellado, CF Arechiga-Flores, H Salinas, JM Luginbuhl
Pages: no - no
Abstract: Puberty is the result of a dynamic interaction between genetic factors and environmental cues, all of which lead to the attainment of reproductive capacity. Thus, significant changes in hormone secretion occur from the pre-pubertal to the pubertal stage. The objective of this review is to provide an update of some endocrine, physiological, metabolic and genetic concepts involved in the establishment of the hypothalamic-hypophyseal-gonadal axis function promoting the onset of the reproductive function during puberty. To achieve this purpose, basic aspects of the function of the hypothalamic-hypophyseal-gonadal axis, the control of the axis by neurotransmitters and the interaction between reproductive function and metabolic status will be considered. Finally, the role of the novel kisspeptin system and the GPR54 receptor as modulators of puberty will be considered, in addition to the hierarchical expression of the main genes acting as regulators of the onset of puberty.
- Comparison of Two Different Programmes of Ovulation Synchronization in the
Treatment of Ovarian Cysts in Dairy Cows
- Authors: N Gundling; S Drews, M Hoedemaker
Pages: no - no
Abstract: It was the aim of this field study to evaluate two different protocols of ovulation synchronization for the treatment of ovarian cysts and their effect on reproductive performance in dairy cows. In addition, factors with a possible influence on treatment success and pregnancy outcome as well as costs per pregnancy were analysed. The study was performed with 130 German Holsteins with ovarian cysts diagnosed on days 55 to 60 postpartum. Cows belonging to group 1 (n = 65) received a modified ovsynch protocol [day 0: 0.15 mg cloprostenol (PGF) + 0.02 mg buserelin acetate (GnRH); day 14: PGF; day 16: GnRH]. Group 2 (n = 65) was treated with the conventional ovsynch protocol (day 0: GnRH; day 7: PGF; day 9: GnRH). Timed artificial insemination was performed 20 to 24 h later. Cows without ovarian cysts served as controls. Treatment success (disappearance of the ovarian cyst) after the first ovsynch cycle was higher in group 1 (66.2%) than in group 2 (23.1%, p
- Luteal Characteristics and Progesterone Production on Day 5 of the Bovine
- Authors: MP Green; GE Mann, MG Hunter
Pages: no - no
Abstract: In this study we have examined luteal function in non-lactating and late lactation dairy cows on day 5 of the cycle, during the period of the post-ovulatory progesterone rise. Comparison of luteal progesterone content and in vitro synthetic capacity with circulating plasma progesterone demonstrated that circulating progesterone concentration is a function of total luteal activity rather than the activity of individual units of tissue. Incubation of luteal tissue in vitro demonstrated stimulatory activity of LH and IGF-I, and to a greater degree IGF-II, on luteal progesterone synthesis. Finally the study showed no effect of double ovulation on luteal function. Occurrence of double ovulation in 35% of animals was not associated with any difference in luteal function or plasma progesterone concentrations.
- Successful Low Dose Insemination of Flow Cytometrically Sorted Ram
Spermatozoa in Sheep
- Authors: SP De Graaf; G Evans, WMC Maxwell, JA Downing, JK O'Brien
Pages: no - no
Abstract: The fertility of ram spermatozoa that had undergone flow cytometric sorting (MoFlo® SX) and cryopreservation was assessed after low-dose insemination of synchronized Merino ewes. Oestrus was synchronized with progestagen-impregnated pessaries, PMSG and GnRH treatment. Ewes (n = 360) were inseminated with 1 × 106, 5 × 106 or 15 × 106 motile sorted frozen-thawed (S1, S5, or S15 respectively) or non-sorted frozen-thawed (C1, C5 or C15 respectively) spermatozoa from three rams. An additional group of ewes were inseminated with 50 × 106 motile non-sorted frozen-thawed spermatozoa (C50) to provide a commercial dose control. The percentage of ewes lambing after insemination was similar for C50 (24/38, 63.2%), C15 (37/54, 68.5%), S15 (38/57, 66.7%), S5 (37/56, 66.1%) and S1 (32/52, 61.5%) groups (p > 0.05), but lower for C5 (19/48, 39.6%) and C1 (19/55, 34.5%) treatments (p
- Bovine Endothelial Cells Interact with Fully-luteinized, but Not
Luteinizing, Granulosa Cells in the mRNA Expression of Endothelin-1 System
in Response to Prostaglandin F2α
- Authors: K Shirasuna; S Watanabe, D Yamamoto, M Hayashi, K Nagai, A Miyamoto
Pages: no - no
Abstract: The corpus luteum (CL) undergoes regression by prostaglandin (PG)F2α from uterus and endothelin-1 (ET-1) plays an important role during luteolysis as a local mediator of PGF2α in the cow. Endothelial cells (EC) and luteal cells are main cell types making up the CL and their interactions are vital for CL function. We aimed to examine the relevance of interactions between EC and luteal cells on stimulation of genes which involved ET-1 synthesis by PGF2α. We further focused the impact of maturity of luteal cells on the stimulation of the genes. To make a microenvironment which resembles the CL, we used bovine aortic endothelial cells (BAEC) and luteinizing or fully-luteinized granulosa cells (GC) and evaluated the effect of PGF2α on the expression for mRNA of ET-1 system by using real-time RT-PCR. PGF2α stimulated the expression of preproET-1 and endothelin converting enzyme-1 mRNA only in the co-cultures of BAEC with fully-luteinized GC, but not with luteinizing GC. The data suggest that interactions between BAEC and fully-luteinized GC enhance the capability of BAEC to produce ET-1 in response to PGF2α. This mechanism may contribute to the local induction of luteolytic action of PGF2α which is dependent on the age/maturation of the CL.
- Effect of Time for Mating and Gestation Length on Reproductive Efficiency
- Authors: Y Shimatsu; H Yuzawa, K Aruga, M. Nakura
Pages: no - no
Abstract: The purpose of this study was to examine the effect of time for mating and gestation length on reproductive efficiency in dogs. Groups of eight, six and six beagle bitches were mated with a total of three sires on days 3, 5 and 7, respectively, after the luteinizing hormone (LH) surge. All the bitches whelped successfully. The gestation lengths (the intervals from the LH surge to the whelping) were 65.1 ± 1.9, 65.5 ± 1.9 and 68.0 ± 1.8 days, respectively. This length of mating 7 days after the LH surge was significantly longer than that of mating 3 and 5 days after the LH surge (p
- Ovum Pick-up in Dairy Heifers: Does it Affect Animal Well-being?
- Authors: S Petyim; R Båge, A Madej, B Larsson
Pages: no - no
Abstract: The aim of the present study was to assess the effects of the ovum pick-up (OPU) technique on animal well-being. Eight dairy heifers were subjected to 4 months of twice-weekly OPU. The physiological response to OPU was recorded in four heifers at two sessions, at the beginning (time 1) and at the end (time 2) of the 4-month period. Heart rates were measured and blood was analysed for cortisol, vasopressin and PG-metabolite before, during (every 5 and 2 min), and after the OPU sessions. Reactions to each subprocedure of OPU (‘restraint’, ‘epidural’, ‘device in’ and ‘puncture’) were closely observed. In all heifers, reactions to the OPU procedures were also noted throughout the experimental period, and changes in routine behaviour, oestrous behaviour, body temperature, or other clinical traits were recorded. Subsequent to the experiment, the ovaries and tails were carefully inspected. At time 1, there was an insignificant increase in heart rate and cortisol throughout the OPU procedure. At time 2, these two parameters increased significantly, but both parameters declined to pre-OPU levels 10 min after completion of the procedure. No significant changes were seen in vasopressin or PG-metabolite at time 1 and time 2. Behaviourally, the heifers showed the strongest response to epidural anaesthesia, with a tendency for more intense response during the late 4-month sessions. The response to ‘device in’ and ‘puncture’ varied among individuals independently of time. There were no changes in the routine or oestrous behaviour throughout the experiment and no signs of clinical disorders. No major pathological changes were macroscopically seen in the ovaries and tails subsequent to the 4 months of OPU. In conclusion, the heifers showed a response to OPU, mostly to administration of epidural anaesthesia. However, we demonstrated that epidural anaesthesia can be administered in a way causing less discomfort.
- Early Detection of Pregnancy and Embryonic Loss in Dairy Cattle by ELISA
- Authors: G Gábor; F Tóth, L Ózsvári, Zs Abonyi-Tóth, RG Sasser
Pages: no - no
Abstract: Over a 25-month period 8118 blood samples were assayed for the presence of the serum pregnancy specific-protein B (PSPB) and progesteron (P4) concentrations on three Hungarian large-scale dairy farms. Pregnancy (n = 4085) was checked by BioPRYN assay at 30–36 days post-insemination (PI). Samples from all cows that tested not pregnant and from cows with an optical density (OD) reading in the BioPRYN test that was between 0% and 30% above the cutoff OD value were tested for serum P4 concentration. According to serum P4 concentration, cows were assigned to three categories: high (>4 ng/ml), medium (2–4 ng/ml) and low (
- Infertility in a Dog due to Proximal Cytoplasmic Droplets in the
Ejaculate: Investigation of the Significance for Sperm Functionality In
- Authors: AI Peña; M Barrio, JJ Becerra, LA Quintela, PG Herradón
Pages: no - no
Abstract: A 4-year-old Basque Shepherd male dog was presented for breeding soundness evaluation after the dog failed to impregnate the three bitches he had mated. Clinical examination showed no anomaly of the reproductive system. Semen evaluation showed normal sperm count (640 × 106), 80% had progressively motile spermatozoa, and 96% had morphologically abnormal sperm of which 84% had proximal cytoplasmic droplet and 12% had proximal droplet plus other anomaly. A zona pellucida-binding assay, using canine oocytes derived from frozen-thawed ovaries, was performed in order to investigate the zona-binding ability of dog spermatozoa with proximal cytoplasmic droplets. For the zona pellucida-binding assay, ovaries were thawed and minced in phosphate-buffered saline + 0.4% bovine serum albumin, the oocytes recovered were divided into two groups of 35–40 oocytes to be, respectively, used with the infertile dog and with a control fertile dog. Spermatozoa were capacitated in Canine Capacitating Medium (CCM) at 38.5°C and 5% CO2 in air for 2 h before oocyte insemination. Groups of five to six oocytes placed in 45 μl droplets of CCM were incubated for 1 h. Afterwards, 5 μl of CCM containing 25 000 spermatozoa were added to each droplet and co-incubated for 2 h before fixation and evaluation of the complexes. After oocyte insemination, sperm motility and viability were evaluated: the sample from the infertile dog had 85% sperm motility with fast and linear progressive movement, and sperm viability of 92%. The sample from the control dog showed 40% sperm motility with fast and highly curvilinear and erratic movement, high degree of sperm agglutination and sperm viability of 32%. For the infertile dog the mean number of bound spermatozoa/oocyte was 0.33 whereas for the control dog it was 1.80. It was concluded that dog sperm with proximal cytoplasmic droplets seem to lack normal capacitating ability in vitro, and consequently, they may have reduced capacity to bind to the zona pellucida of canine oocytes.
- Distribution of Spermatozoa and Embryos in the Female Reproductive Tract
after Unilateral Deep Intra Uterine Insemination in the Pig
- Authors: P Tummaruk; P Sumransap, M Techakumphu, A Kunavongkrit
Pages: no - no
Abstract: The present study was performed to investigate the number of either the spermatozoa or the embryos in the reproductive tracts of sows after unilateral, deep, intra uterine insemination (DIUI). Two experiments were conducted, 10 sows were used in experiment I and eight sows were used in experiment II. Transrectal ultrasonography was used to examine the time when ovulation took place in relation to oestrus behaviour. The sows were inseminated with a single dose of diluted fresh semen 6–8 h prior to expected ovulation, during the second oestrus after weaning. In experimental I, five sows were inseminated by a conventional artificial insemination (AI) technique using 100 ml of diluted fresh semen, containing 3000 × 106 motile spermatozoa and five sows were inseminated by the DIUI technique with 5 ml of diluted fresh semen, containing 150 × 106 motile spermatozoa. The sows were anesthetized and ovario-hysterectomized approximately 24 h after insemination. The oviducts and the uterine horns on each side of the reproductive tracts were divided into seven segments, namely ampulla, cranial isthmus, caudal isthmus, utero-tubal junction (UTJ), cranial uterine horn, middle uterine horn and caudal uterine horn. Each segment of the reproductive tracts was flushed with Beltsville thawing solution (BTS) through the lumen. The total number of spermatozoa in the flushing from each segment were determined. In experimental II, eight sows were inseminated by the DIUI technique using 5.0 ml diluted fresh semen containing 150 × 106 motile spermatozoa. The sows were anesthetized 61.1 ± 12 h after insemination (48–72 h) and the embryos were flushed from the oviduct through the proximal part of the uterine horn. It was revealed that, in experimental I, the spermatozoa were recovered from both sides of the reproductive tract in the AI-group, and from unilateral side of the reproductive tract in the DIUI-group (three sows from the left and two sows from the right sides). The number of spermatozoa recovered from the reproductive tracts was higher in the AI- than the DIUI-group (p