Infertile Animals Research Articles

Preclinical examination of complex antimicrobial preparation for treating endometritis in cows

According to a statistical study an average of 40 % of infertile animals have obstetric and gynecological pathologies. Among obstetric and gynecological pathologies, the most common is an acute postpartum endometritis, which make up from 37.8 to 41 %. Therapy and Pharmacology department of the Kuban State Agrarian University has developed a drug for the cows postpartum endometritis treatment. Conducted preclinical tests prove its harmlessness, as a result of which it belongs to hazard class 4. It should be noted that the biochemical blood parameters of rats in the experiment do not go beyond the reference values. The drug has a pronounced antimicrobial effect. MBC is from 7.8 mg to 31.2 mg while MIC is from 31.2 to 62.5 mg for such strains as S. aureus, E. coli, St. xylosus, Ps. aeruginosa, P. vulgaris. The developed drug does not have an irritating and sensitizing effect and exhibits a regenerative effect. Conducted experiments on the wound healing effect of the drug prove that this drug has a high regenerative effect. The average wound healing area was 20.4 % per day.

Clinical and laboratory prognostic indicators for fertility in sheep

With the use of technology of artificial insemination in sheep farms are not yet defined criteria for assessing the full value of the preparation of females for insemination, and hence – the possibility of prediction and correction of their fertilization, which prevents the rational use of cryopreserved semen and ensuring the maximum reception of the offspring. We have proved the prognostic importance of fertilization of the morphofunctional state of the vulva and the vagina and the quality of mucus in sheep breeds during estrus. The material of the study was 327 sheep of ascanian breeding, which during the sexual intercourse before insemination examined the vulva, vagina and evaluated the estrus slime (number, color, consistency, presence of impurities, elasticity, electrical resistance, type of crystallization, protein content). The results of ultrasound diagnosis of pregnancy were determined by the fertility of sheep with different integral compositions of clinical and laboratory parameters. It has been established that in sheep, in which fertility in the first sexual cycle reached 65.1%, most often found a pink, moderately edematous vulva with clear mucus. In females with pale mucous membrane and insignificant amount of mucus, the effectiveness of inseminates declined to 53.3-58.0%. In a significant amount of estral mucus, the proportion of infertile animals increased twofold. At the same time, the selection of liquid, but cloudy, or thick mucus is a sign of an unfavorable prognosis, in which fertility decreases by 1.8-2.1 times (p <0.001). Dense, white, paste-like isolates were observed in a small number of sheep, mostly bright at the beginning of the anestral season. Low fertility in the first sexual hunting (35.5%) and a high multiplicity of repeated inseminations (29.0%) indicate that sheep with thick estral slime are only beginning to enter the sexual season, and this quality of secrecy indicates an inadequate estrogenization of the body. It was also found that in the infertile sheep during sexual hunting, the protein content of cervical mucus was 4.8 times higher, and the elasticity of mucus was reduced by 2.9 times. The most prevalent was the prognosis of average fertilization (53.3-58.0%), which was recorded in 62.9% of experimental sheep. The prognosis of high fertilization, which resulted in 62.5-65.1% of oseminins, was found in 27.8% of females. At the same time, the number of females with a fertility forecast at 40% was only 3.1%, and the prevalence of an unfavorable prognosis, in which fertility was the smallest (30.0-35.5%), reached 6.2%. Key words: sheep, askanian breeding, estrus, fertility prognosis, estral mucus, vulva, vagina, artificial insemination.

Nutritional Infertility and Ameliorative Measures in Dairy Animals of Middle Gujarat

Fertility improvement camps were organized in different villages of middle Gujarat covering 1481 breedable cattle and buffaloes with the history of impaired fertility. The reproductive status of animals was ascertained through gynaeco-clinical examination per rectum. Blood sampling (n=305) was done from representative groups of animals to know the nutritional status. All non-infectious infertile animals were dewormed using either Inj. Ivermectin 100 mg s/c or bolus Fenbendazole + Ivermectin 3.0 g orally. Anestrus animals (n=265) were then nutritionally supplemented either with chelated area specific mineral mixture (ASMM, 50-60 g/d/h) or multi-minerals boli - one on alternate day, with and without bypass fat (75-80 g/d/h) for one month along with an advice to improve feeding level, estrus detection and timely breeding. Revisits were made after 2.5-3.0 months to check the reproductive status. The mean blood glucose concentrations were significantly lower (p less than 0.05), while total cholesterol and zinc levels were higher (p less than 0.05) in anestrus than in subestrus and/or repeat breeding cows and buffaloes. Moreover, the plasma total cholesterol levels in buffaloes were just one half to those of cows. Further, the plasma total protein, calcium, phosphorus, magnesium, iron, copper and cobalt concentrations were found to be nonsignificantly higher in repeat breeders than in anestrus or subestrus cows and buffaloes. Calcium was deficient in some 20% animals and calcium: phosphorus ratio varied from 1:1 to 1.8:1 in animals under different areas of study indicating its role in causing infertility in dairy animals. Supplementation of ASMM, multi-mineral boli alone or in combination with bypass fat in pubertal /postpartum anestrus animals resulted in 34-56% estrus induction response and 33-50% conception rate, both being better with mineral plus bypass fat supplementation than minerals alone.

The use of new technologies in horse selection

Received: 2018-09-12 | Accepted: 2018-11-27 | Available online: 2018-12-31 https://doi.org/10.15414/afz.2018.21.04.186-189 Most successful selection strategies are not in use in horse selection. This study describes the use of current strategies of selection in horse and tries to answers why well known, scientifically proven selection strategies are still not implemented. In the era of genomic selection (GS) in animal breeding, decision making in horse selection, even classical values (BV), are not fully taken advantage of. Breeding organizations (BO) which implement BVs as selection criteria in their programs show great success in genetic gain. However, a horse's achievements and the popularity of its relatives still plays an important role in selection level. Genomic based selection tools in horse have limitations in quantitative traits since it is difficult to establish reference population. Mendelian traits have been studied for several years, especially those related to horse health and functional longevity (long sport or race careers), yet there are still only a small number of validated mendelian traits offered for horses. The important benefit of GS is the prevention of related mating based on genomic data, in addition to pedigree data. The specialty in horse is the use of cloning. From the point of view of genetic variability, cloning is useful in enabling geldings or non-genetically infertile animals to carry genes to next generations. In sport and race BOs, stallions from foreign BOs are frequently used. In such cases a need for comparable BVs exists. These factors were the motivation behind the establishment of Interstallion 20 years ago. Due to difficulties in BO collaboration, Interstallion has not been as successful as planned. On the whole, it is expected that the sector of horse selection will need to change considerably in the future. Keywords: value, cloning selection, genomic selection, inbreeding, strategies References Arnason, T. (1984) Genetic studies on conformation and performance of Icelandic toelter horses: IV Best linear unbiased prediction of ten correlated traits by use of an animal model. Acta Agric. Scand. 34, 450–462. Broere Jazz (2018) Jazz – one in a million. [Online] Alblasserdam. Available at: http://www.broerejazz.com/en/sperma-bestellen/ [Accessed 12 August 2018]. FMITCHELL07 (2009) Inbreeding to broodmares: an in-depth statistical analysis of the Rasmussen Factor, Part 1 [Online] https://fmitchell07.wordpress.com/2009/07/29/inbreeding-to-broodmares-an-in-depth-statistical-analysis-of-the-rasmussen-factor-part-1/ [Accessed 12 August 2018]. Henderson, C. R. (1953) Estimation of variance and covariance components. Biometrics, 9, 226–252. Jorjani, H., Jakobsen, J., Hjerpe, E., Palucci, V., Durr, J. (2012) Status of genomic evaluation in the Brown Swiss populations. Interbull Bull., 46, pp. 46–54. Koenen, E. (2005) Publication of values: Interstallion guidelines. Workshop Use of genetic evaluations in sport horse breeding Warendorf, Germany. [Online] In: Leuven: KU Leuven. Available at: https://www.biw.kuleuven.be/Genlog/livgen/research/interstallion/workshop_docs/koenen1.pdf [Accessed 12 August 2018]. Koenen, E., Aldridge, L.I., Philipsson, J. (2004) An overview of objectives for warmblood sport horses. Livestock Production Science, 88(1-2), pp. 77-84. Liljenstolpe, C. (2009) Horses in Europe. [Online] Maarslet: WBFSH. Available at: http://www.wbfsh.org/files/EU%20Equus%202009.pdf [Accessed 16 August 2018]. Mark, T. (2013) Genomic selection for performance and health http://www.wbfsh.org/files/Thomas_Mark_Genomic_Selection_in_Horse_Breeding_Oct._7th_2013_Warsaw.pdf [Accessed 12 August 2018]. OMIA (2018) Online Mendelian Inheritance in Animals (OMIA). [Online] Maarslet: WBFSH. Available at: http://omia.org/home/ [Accessed 6 September 2018]. Philipsson, J. (2009) Mace for horse evaluations. Interstallion: Workshop Le Pin au Haras, 26th March 2009. [Online. Available at: ] https://www.biw.kuleuven.be/Genlog/livgen/research/interstallion/workshop_LePin/I_MACE%20for%20horse%20evaluations.pdf [Accessed 12 August 2018]. Reed, T. (2018) Johan Knaap on genomic selection – an interview. [Online]. Available at: http://www.irish-warmblood.com/pdf/Johan_Knaap_on_Genomic_Selection.pdf [Accessed 6 September 2018]. Reis, A.P., Palmer, E., Nakhla, M. (2012) Biotechnologies of reproduction in the horse: what has changed? In: Book of Abstracts of the 63 rd Annual Meeting of the European Federation of Animal Science Wageningen. Wageningen: Wageningen Academic Publishers. Shelbourne Farm. (2018) Cruising Clones. [Online]. Available at: http://www.shelbournefarm.com/stallions/cruising-clones/ [Accessed 6 September 2018]. Stock, K., Quinn Brady, K., Christiansen, K., Viklund, A., Cervantes, I., Ricard, A., Ducro, B., Janssens, S. (2015). Breeding objectives and practices of sport horse studbooks : Results of a worldwide inventory. In: Book of Abstracts of the 66 th Annual Meeting of the European Federation of Animal Science Wageningen.Wageningen: Wageningen Academic Publishers. Stock, K., Viklund, A., Cervantes, I., Ricard, A., Christiansen, K., Vangen, O., Janssens, S. (2018). The role of Interstallion in supporting international sport horse by improved transparency. In: Book of Abstracts of the 69 th Annual Meeting of the European Federation of Animal Science Wageningen. Wageningen: Wageningen Academic Publishers. Thoren Hellsten, E. (2008) International Sport Horse Data for Genetic Evaluation. Doctoral Thesis. Uppsala: Swedish University of Agricultural Sciences. Waikato (2018) Waikato Stud offers a truly world-class stallion line up. [Online]. Available at: https://waikatostud.com/stallions/ [Accessed 6 September 2018]. WBFSH (2018) World federation for sport horses: Breeder and Studbook rankings. [Online]. Available at: http://www.wbfsh.org/GB/Rankings/Breeder%20and%20Studbook%20rankings.aspx [Accessed 12 September 2018].

Effect of platelet-rich plasma (PRP) on ovarian structures in cyclophosphamide-induced ovarian failure in female rats: a stereological study

Background: Ovarian failure is diagnosed by ovarian destruction and reducing sex hormonal levels. Platelet-rich plasma (PRP) contains several growth factors that induce tissue repair and may induce folliculogenesis.Objective: This study evaluates the effect of PRP on ovarian structures and function in cyclophosphamide (Cy)-induced ovarian failure in female rats by a stereological method.Methods: Thirty-two adult female rats were divided into four groups (Control, Cy, Cy + PRP, and PRP). Female infertility was induced by Cy (75 mg/kg, single dose, intraperitoneally). Animals were treated by PRP which was obtained from age-matched male rats (200 μL, single dose, intraperitoneally). Blood samples were collected for measurement of hormones. The animals were dissected and the right ovaries were removed, fixed, sectioned, and stained by H&E. Stereological methods were used to estimate cortex and medulla volume, and the number and diameter of follicles, follicular cell, and oocyte using light microscopy.Results: Cyclophosphamide had the maximum effect in decreasing on cortex volume, the pre-antral follicles number, a diameter of follicular cells and oocyte diameter in the antral follicle and the reduction of estradiol and progesterone levels compared with the control group. PRP had a dominant positive effect on the ovarian cortex volume, pre-antral follicles number and antral follicle diameter relative to the control group. PRP also decreased oocyte diameter in pre-antral follicle in infertile animals (p < 0.001).Conclusion: It seems that PRP has a protective effect on ovarian failure in the infertile female rat model.

Polymorphisms of the Bovine Bone Morphogenetic Protein 7 Gene (BMP 7) and its Association with Infertility in Egyptian Water Buffaloes

BMP 7 is a member of transforming growth factor-β (TGF- β) superfamily which plays an important role in the growth and follicular development in mammalian ovary. The present study was designed to amplify and detect polymorphism in exon 6 and exon7 of BMP 7 gene and to investigate the association with infertility in Egyptian buffalo cows. PCR amplification of DNA isolated from blood of 35 Egyptian water buffalo cows (5 normal fertile, 16 repeat breeders and 14 anestrum) revealed the expected size (259 bp) for exon 6 and (360 bp) for (Part of intron 6 plus part of exon 7). Polymorphisms in these two loci were studied by using direct sequencing and RFLP techniques. No SNPs were detected in exon 6, a monomorphic restriction banding pattern observed in normal cyclic and infertile animals. Non- synonomous SNP (G/A) at base 82 from the start of exon 7 was discovered in BMP7 gene. G to A transition revealed two genotypes GG and GA in the investigated Egyptian water buffaloes. The GG genotype was predominant with a genotype frequency of 0.94. GA genotype was found in some infertile animal with frequency of 0.06. SNP (G82A) of BMP7 exon7 gene might be useful in marker-assisted selection of breeding programs for the Egyptian water buffalo.

Studies on ruminant brucellosis in El Salam canal area, Egypt

The current study is aimed to provide some epidemiological data about brucellosis and its impact among ruminants in the area around El-Salam canal, North Sinai. In addition to determination of the sensitivity, specificity; positive & negative predictive values and diagnostic efficacy of Brucella Antibody Test Kits as a rapid field assay for the diagnosis of brucellosis. Our results revealed that, the overall prevalence rate of brucellosis was 7.1% (12/168) and 10.1% (17/168) using ELISA and Brucella Antibody Test Kits respectively. Out of 168 serum samples; 9 were seropositive and 148 were sero-negative for brucellosis by both ELISA and Brucella Antibody Test Kits. Another 3 and 8 serum samples were only positive with ELISA and Brucella Antibody Test Kits respectively, revealing 75% sensitivity, 94.8% specificity, 47% positive predictive value, 53.5% negative predictive value and 93.4% diagnostic efficacy for this test. Brucellosis was found to be one of the causes of abortion in the study area as 61.9% (13/21) of aborted animals were seropositive for brucellosis. While to a little extent, it can be considered one of infertility causes as 4.7% (7/147) of infertile animals were seropositive for brucellosis. On the other hand, there were 38% (8/21) and 95.2% (140/147) of aborted and infertile animals respectively were sero-negative for brucellosis revealing that, there were another causes of abortion and infertility in the area of study rather than brucellosis and there was a necessity for their further investigations in order to improve the fertility and productivity of the animals in this targeted area.

Artificial Induction of Lactation in Bovines: Scope and Limitations

Estrogen, progesterone, prolactin and glucocorticoids work synergistically at the onset of lactogenesis. Estrogen stimulates mammary ductal growth, and the combination of estrogen and progesterone regulates lobular-alveolar development of the mammary gland. This development does not occur in infertile animal or animals suffering from reproductive disorders due to relative absence of sex steroid hormones. Artificial induction of lactation can be induced in such animals by the combination of steroid hormones estrogen and progesterone. Additionally reserpine (tranquilizer- increases prolactin level) and dexamethasone (a synthetic glucocorticoid) can be used in induction protocols to increase the success rate. Artificial induction reduces the involuntary culling rate as well as the genetic losses. To save such genetically superior bovines, induction of lactation is the most appropriate technology used under Indian conditions. The treatment is more effective in well-fed animals with good body condition score and having normal teat and udder. Hormonal induction of lactation should be practice only in infertile animals; it should not be followed as a routine practice, as an alternate to normal lactation. Estrogen and progesterone in 1:2.5 ratios is effective in exotic and cross bred while 1:1 ratio is effective in indigenous cows and buffaloes for artificial induction because of short luteal phase. These methods of artificial induction of lactation yield 60 to 70% milk of previous lactation.

333 GENERATION OF OOCYTE-LIKE STRUCTURE FROM OVARIAN SURFACE EPITHELIAL STEM CELLS OF GOAT

Stem cells have potential for therapeutic application. Continuous repair of ovarian surface epithelium following folliculogenesis and ovarian carcinoma suggests the presence of stem cells in ovarian epithelial cells. In vitro gametogenesis in livestock will result in large numbers of oocytes production from a single ovary, resulting in faster multiplication of superior germplasm of livestock species, treatment of infertile animals, and conservation of endangered species. The present study was conducted with the objective of in vitro differentiation of putative ovarian surface epithelial stem cells into oocyte-like structures in goat model. Ovary samples of 1- to 2-year-old goats were collected from slaughterhouse. The surface of the ovary was gently scraped using sterile blunt scraper to isolate ovarian surface epithelial stem cells. These scraped cells were cultured in DMEM/F12 supplemented with 20% FBS for 3 weeks in 5% CO2 at 37°C with maximum humidity. The cultured stem cells were characterised for stemness by RT-PCR and immunostaining for Oct4, Sox2, and Nanog genes after 3 weeks. These putative stem cells were in vitro differentiated spontaneously to oocyte-like structures in DMEM/F12 medium and characterised for premeiotic markers by RT-PCR and immunostaining for VASA, DAZL, and STELLA genes. Results of this study provide evidence for the presence of putative stem cells with pluripotent characteristics in the ovarian surface epithelium. The cultured cells were found to be round in shape, with a high nucleus to cytoplasm ratio under inverted microscope, and found positive for stem cell markers of Oct4, Sox2, and Nanog genes. A total of 66 oocyte-like structures were produced from 12 ovaries. These oocyte-like structures were nearly similar to oocytes produced in vivo, both morphologically and in molecular gene expression. The oocyte-like structures were also found positive for premeiotic markers of VASA, DAZL, and STELLA genes by RT-PCR and immunostaining. From this study, we concluded that the ovarian surface epithelial cells have putative stem cells which can be in vitro differentiated into oocyte-like structures in goat. These oocyte-like structures need further characterisation of their surface membrane, more molecular markers, and following their developmental potential. These oocytes can help for multiplication of elite germplasm, curing infertile animals, and saving endangered species.

СВОБОДНОРАДИКАЛЬНОЕ ОКИСЛЕНИЕ ЛИПИДОВ И РЕПРОДУКТИВНОЕ ЗДОРОВЬЕ КОРОВ

Free radical lipid oxidation is currently considered as one of the dominant metabolic processes of physiological system functional activity. When it goes beyond regulated limits it is considered as an inductor of free radical pathology oxidative stress. In the conditions of a big dairy cattle breeding farm (Voronezh Province), specialized in Red-and-White breed, the functioning peculiarities of the peroxidation system lipid-oxidant defense in cows at the normal course of gestation and at gestosis, at the normal course of postpartum period and at puerperal endometritis, uterus subinvolution and ovarian dysfunction were studied. The state of lipid peroxidation processes and antioxidant system were evaluated by determining total lipid content, cholesterol, triglycerides, malonic dialdehyde (MDA), nitrogen oxides (NO ·), E and C vitamins concentrations, glutathione peroxidase (GPO), glutathione reductase (GR), catalase, superoxide dismustase (SOD) activity in blood. It is demonstrated that the high activity of lipid peroxidation reactions and the system of nitrogen oxide at decrease of antioxidant defense non-enzymic link capacity is the basis of gestosis and acute postnatal complications. Thus, аn increase of MDA concentration by 42.3-43.0 %, NO · by 31.9-38.0 % was observed in cows with gestosis. At mild gestosis the glutathione peroxidase activity, catalase activity and vitamin C concentration increased by 11.0 %, 14.3 %, 38.0 %, and 24.1 %, respectively, while vitamin Е concentration decreased by 11.7 % due to more consumption for neutralization of the toxic peroxidation products. At more sever pathology, the glutathione peroxidase activity and catalase activity in blood increased by 26.0 % and 17.3 %, respectively, when compared to the healthy animals, while vitamins E and C concentrations decreased by 33.3 % (р < 0,01) and 17.2 %, respectively. As free radical oxidation intensified, an anaerobic degradation of carbohydrates was activated to supply the tissues of developing fetus with energy under oxygen deficit occurred because of violation of the blood circulation. Postpartum inflammation in the genital organs in cows developed against the background of increase of MDA concentration by 76.0 %, GPO and GR activity by 65.8 % and 14.6 %, respectively, SOD by 46.0 %, catalase by 45.7 %, 2.9 times increase of NO · concentration and reduction of vitamin E content by 35.5 %. Infertile animals with ovarian dysfunction were characterized by high activity of lipid peroxidation processes and by low level of nitrogen oxide generation. This is indicated by an increased concentration of MDA by 57.0 %, activity of GPO by 27.6 %, GR by 10.5 %, SOD by 31.9 %, catalase by 24.3 %, with a reduced content of NO · and vitamin E by 56.9 % and 31.6 %, respectively, in comparison with healthy animals. A decrease in NO · concentration in blood could result from a sharp depression of hormone synthesizing function in ovaries, and low NO · production could disturb functions of the gonads. The high level of peroxidation is peculiar to animals with chronic uterus pathology, however, it is less expressed than in cows with acute course of the pathological process.

FREE RADICAL LIPID OXIDATION AND REPRODUCTIVE HEALTH OF COWS

Free radical lipid oxidation is currently considered as one of the dominant metabolic processes of physiological system functional activity. When it goes beyond regulated limits it is considered as an inductor of free radical pathology oxidative stress. In the conditions of a big dairy cattle breeding farm (Voronezh Province), specialized in Red-and-White breed, the functioning peculiarities of the peroxidation system lipid-oxidant defense in cows at the normal course of gestation and at gestosis, at the normal course of postpartum period and at puerperal endometritis, uterus subinvolution and ovarian dysfunction were studied. The state of lipid peroxidation processes and antioxidant system were evaluated by determining total lipid content, cholesterol, triglycerides, malonic dialdehyde (MDA), nitrogen oxides (NO • ), E and C vitamins concentrations, glutathione peroxidase (GPO), glutathione reductase (GR), catalase, superoxide dismustase (SOD) activity in blood. It is demonstrated that the high activity of lipid peroxidation reactions and the system of nitrogen oxide at decrease of antioxidant defense non-enzymic link capacity is the basis of gestosis and acute postnatal complications. Thus, an increase of MDA concentration by 42.3-43.0 %, NO • by 31.9-38.0 % was observed in cows with gestosis. At mild gestosis the glutathione peroxidase activity, catalase activity and vitamin C concentration increased by 11.0 %, 14.3 %, 38.0 %, and 24.1 %, respectively, while vitamin A concentration decreased by 11.7 % due to more consumption for neutralization of the toxic peroxidation products. At more sever pathology, the glutathione peroxidase activity and catalase activity in blood increased by 26.0 % and 17.3 %, respectively, when compared to the healthy animals, while vitamins E and C concentrations decreased by 33.3 % (ð < 0,01) and 17.2 %, respectively. As free radical oxidation intensified, an anaerobic degradation of carbohydrates was activated to supply the tissues of developing fetus with energy under oxygen deficit occurred because of violation of the blood circulation. Postpartum inflammation in the genital organs in cows developed against the background of increase of MDA concentration by 76.0 %, GPO and GR activity by 65.8 % and 14.6 %, respectively, SOD by 46.0 %, catalase by 45.7 %, 2.9 times increase of NO • concentration and reduction of vitamin E content by 35.5 %. Infertile animals with ovarian dysfunction were characterized by high activity of lipid peroxidation processes and by low level of nitrogen oxide generation. This is indicated by an increased concentration of MDA by 57.0 %, activity of GPO by 27.6 %, GR by 10.5 %, SOD by 31.9 %, catalase by 24.3 %, with a reduced content of NO • • •

Evaluation of the breeding soundness of male camels (Camelus dromedarius) via clinical examination, semen analysis, ultrasonography and testicular biopsy: a summary of 80 clinical cases.

Male camel infertility is a heterogeneous disorder. A variety of factors may adversely affect sperm production and function and impair fertility. This study was designed to evaluate the sensitivity and specificity of ultrasonography and testicular biopsy in the evaluation of the breeding soundness of male dromedaries compared with results obtained by clinical examination and semen analysis. Eighty-four male dromedary camels (5-15years old) were used in this study during the rutting season (November-May). Four sexually mature male camels were used as controls. These animals were apparently healthy and had histories of normal fertility. Eighty infertile male camels were subjected to an algorithmic approach based on information collected during careful examinations of the camels' breeding histories, clinical examinations, testicular evaluations, testicular ultrasonographies, the results of the semen analyses and testicular biopsies to diagnose the camels' infertilities. The differences in the semen parameters between the control and infertile male camels were highly significant (p<0.01). Regarding the diagnoses of male camel infertility, the results of testicular ultrasonographies and biopsies were compared with those from the semen analyses, and the accuracies of these tests were 92.5% and 90%, respectively. Additionally, the results of the testicular ultrasonographies were matched with those of the testicular biopsies of the infertile animals, and this comparison resulted in 85% accuracy. Testicular biopsy is a promising method that, along with a carefully performed history, clinical examination, an appropriate testicular ultrasonography procedure and semen analysis, can afford veterinarians the opportunity for more precise diagnosis and treatment of many dromedary infertility disorders.

Evaluación de la aptitud reproductiva del toro

&lt;p&gt;La evaluación de la aptitud reproductiva del toro es un examen sencillo, práctico y económico que permite identificar animales subfértiles e infértiles que pueden llegar a afectar el éxito de un programa reproductivo; en consecuencia, debería realizarse de manera periódica, sin embargo, su utilización en nuestro país es baja. Esta evaluación involucra tres pasos fundamentales: el examen físico (conformación general, órganos sexuales internos y externos), la evaluación de la libido y la colecta y evaluación de semen. Se realiza una revisión de los aspectos que contemplan la evaluación reproductiva del toro, con el fin de determinar la aptitud reproductiva de los machos destinados para tal fin.&lt;/p&gt;

Cell-cycle-dependent Colonization of Mouse Spermatogonial Stem Cells After Transplantation into Seminiferous Tubules

Spermatogonial stem cells (SSCs) migrate to the niche upon introduction into the seminiferous tubules of the testis of infertile animals. However, only 5–10% of the transplanted cells colonize recipient testes. In this study, we analyzed the impact of cell cycle on spermatogonial transplantation. We used fluorescent ubiquitination-based cell cycle indicator transgenic mice to examine the influence of cell cycle on SSC activity of mouse germline stem (GS) cells, a population of cultured spermatogonia enriched for SSCs. GS cells in the G1 phase are more efficient than those in the S/G2-M phase in colonizing the seminiferous tubules of adult mice. Cells in the G1 phase not only showed higher expression levels of GFRA1, a component of the GDNF self-renewal factor receptor, but also adhered more efficiently to laminin-coated plates. Furthermore, this cell cycle-dependency was not observed when cells were transplanted into immature pup recipients, which do not have the blood-testis barrier (BTB) between Sertoli cells, suggesting that cells in the G1 phase may passage through the BTB more readily than cells in the S/G2-M phase. Thus cell cycle status is an important factor in regulating SSC migration to the niche.

Emerging evidence for the role of genomic instability in male factor infertility

Male infertility is a common and complex pathology affecting0about 7% of men of reproductive age. Given its complexity, the underlying etiology for male infertility is often unknown. A growing amount of evidence suggests genomic instability may be an important factor in some cases of male factor infertility. While some specific manifestations of genomic instability, such as increased sperm aneuploidy rates and increased somatic translocations and inversions in infertile men, are well established, other facets of genomic instability associated with male infertility have not been thoroughly investigated. A limited body of recent work has identified a potential association between microsatellite instability and spermatogenic failure. In addition, mutations in mismatch repair and tumor suppressor genes, which could potentially lead to genomic instability, have been identified in some infertile men and animal models. In addition, results of two epidemiologic studies suggest spermatogenic defects might be just one aspect of a more systemic problem, possibly due to increased genomic instability. In this review we discuss well-established links between genomic instability and male infertility, as well as some of the emerging but less established data to support this relationship. We also propose some important areas of future research toward a more complete understanding of the underlying mechanisms for male infertility.

Developmental Dioxin Exposure Negatively Affects Fertility and Pregnancy Outcomes in Mice for Multiple Generations.

TCDD (2,3,7,8-tetrachlorodibenzo-p-dioxin) is an ubiquitous environmental contaminant and known endocrine disruptor, capable of interfering with multiple aspects of mammalian reproduction. Mammals are most sensitive to toxicant exposure during early development; thus we developed a mouse model of in utero TCDD exposure in order to examine the impact of this toxicant on adult reproductive function. For our study, pregnant C57bl/6 mice were provided a single dose of TCDD (10 μg/kg) by gavage on gestation day 15.5 (E15.5) and the dams were observed until normal delivery. Female offspring (F1) of TCDD exposed dams and several subsequent generations (F2-F4) were examined at sexual maturity for fertility, uterine decidualization capacity and pregnancy outcomes. Our initial studies of F1 animals revealed diminished uterine progesterone receptor (PR) expression and the majority of mice were infertile or exhibited a poor decidualization response leading to early pregnancy failure. Among mice which were able to achieve pregnancy, preterm birth (PTB) was common. Examination of surviving F3/F4 mice also revealed a significant number of infertile animals, and this transgenerational effect of TCDD was also correlated with diminished uterine PR and impaired decidualization. In F3/F4 mice which were able to become pregnant, ancestral toxicant exposure was associated with a significant increased risk of PTB. In order to further evaluate pregnancy failure in toxicant-exposed mice, in a subsequent study, we euthanized both control and F1 mice during late pregnancy (E18.5) for qRT-PCR analysis of selected genes at the maternal-fetal interface. These studies revealed that a history of developmental toxicant exposure was associated with a premature placental loss of PR mRNA expression and a concomitant increase in expression of toll-like receptor-4 mRNA, a mediator of inflammation. Since immunosuppression is a major function of progesterone related to the timing of birth, we speculated that a loss of PR in our toxicant-exposed mice may lead to PTB as a consequence of a heightened inflammatory response. To test this theory, we examined the ability of pregnant F1 and F3 mice to resist an inflammatory challenge mediated by intraperitoneal injection of lipopolysaccharide (LPS). Following the identification of a dose of LPS that did not induce PTB in control mice (200 μg/kg), F1 and F3 mice were subjected to an identical LPS challenge on E15.5 of pregnancy. In contrast to control animals, which were resistant to the LPS challenge, 100% of mice with a history of TCDD exposure (F1) or their descendants (F3) delivered within 16 hrs of treatment. Our data suggest that TCDD exposure during early development affects multiple aspects of reproductive tract function in adult animals. Significantly, following early life TCDD exposure, we observed a multigenerational incidence of PTB associated with increased sensitivity to non-microbial inflammation at the maternal-fetal interface. The rate of PTB in women in industrialized countries continues to increase despite better health care and patient awareness. Our experimental model in mice supports the possibility that ancestral, early life TCDD exposure may lead to heritable epigenetic alterations affecting fertility and birth outcomes. In humans, the risk of PTB may be yet another adult condition with its origins in the fetal environment. Support: NIEHS ES014942, NCCAM AT006245 and The Endometriosis Association (platform)

Genome-wide profiling of gene expression in the epididymis of alpha-chlorohydrin-induced infertile rats using an oligonucleotide microarray

BackgroundAs one of the chlorinated antifertility compounds, alpha-chlorohydrin (ACH) can inhibit glyceraldehyde-3-phosphate dehydrogenase (G3PDH) activity in epididymal sperm and affect sperm energy metabolism, maturation and fertilization, eventually leading to male infertility. Further studies demonstrated that the inhibitory effect of ACH on G3PDH is not only confined to epididymal sperm but also to the epididymis. Moreover, little investigation on gene expression changes in the epididymis after ACH treatment has been conducted. Therefore, gene expression studies may indicate new epididymal targets related to sperm maturation and fertility through the analysis of ACH-treated infertile animals.MethodsRats were treated with ACH for ten consecutive days, and then each male rat copulated with two female rats in proestrus. Then sperm maturation and other fertility parameters were analyzed. Furthermore, we identified epididymal-specific genes that are associated with fertility between control and ACH groups using an Affymetrix Rat 230 2.0 oligo-microarray. Finally, we performed RT-PCR analysis for several differentially expressed genes to validate the alteration in gene expression observed by oligonucleotide microarray.ResultsAmong all the differentially expressed genes, we analyzed and screened the down-regulated genes associated with metabolism processes, which are considered the major targets of ACH action. Simultaneously, the genes that were up-regulated by chlorohydrin were detected. The genes that negatively regulate sperm maturation and fertility include apoptosis and immune-related genes and have not been reported previously. The overall results of PCR analysis for selected genes were consistent with the array data.ConclusionsIn this study, we have described the genome-wide profiles of gene expression in the epididymides of infertile rats induced by ACH, which could become potential epididymal specific targets for male contraception and infertility treatment.

Induced ovulation mimics the time-table of natural development in the stripe-faced dunnart, Sminthopsis macroura and results in the birth of fertile young

Induced ovulation maximizes captive breeding success, increasing productivity and facilitating the contribution of otherwise infertile animals to the genetic pool. In marsupials, induced ovulation to produce fertile young is unknown. Here we present an induction protocol efficient in inducing non-cycling and non-reproductive females to cycle, mate, ovulate, and conceive. Ovulation was induced in Sminthopsis macroura using an initial injection of 0.06 IU equine serum gonadotropin (eSG)/g (time 0), followed on day 4 by 0.04 IU eSG/g. Using this induction regime, the timing of follicular and embryonic development mimics natural cycles and results in the birth of viable, fertile young. Response to induction is not significantly affected by animal age, making this protocol an effective conservation tool. We have established a time-table of development following induction, providing a source of precisely timed research material. This is the first induced ovulation protocol in any marsupial to result in demonstrated fertile offspring and to allow the reliable collection of known-age samples during both the follicular phase and the gestation period.

The Effect of Eurycoma longifolia on Sperm Quality of Male Rats

The present study investigated the effects of a standardized methanol extract of E. longifolia Jack containing the major quassinoid constituents of 13alpha(21)-epoxyeurycomanone (1), eurycomanone (2), 13alpha,21-dihydroeurycomanone (3) and eurycomanol (4) on the epididymal spermatozoa profile of normal and Andrographis paniculata induced infertile rats. The standardized MeOH extract at doses of 50, 100 and 200 mg/kg, the EtOAc fraction (70 mg/kg), and standardized MeOH extract at 200 mg/kg co-administered with the EtOAc fraction of A. paniculata at 70 mg/kg were each given orally to male Sprague-Dawley albino rats for 48 consecutive days. The spermatozoa count, morphology, motility, plasma testosterone level and Leydig cell count of the animals were statistically analyzed by ANOVA with a post-hoc Tukey HSD test. The results showed that the sperm count of rats given the standardized MeOH extract alone at doses of 50, 100 and 200 mg/kg were increased by 78.9, 94.3 and 99.2%, respectively when compared with that of control (p < 0.01). The low count, poor motility and abnormal morphology of the spermatozoa induced by the A. paniculata fraction were significantly reversed by the standardized MeOH extract of E. longifolia (p < 0.001). The plasma testosterone level of the rats treated with the standardized MeOH extract at 200 mg/kg was significantly increased (p < 0.01) when compared with that of the control and infertile animals. The spermatocytes in the seminiferous tubules and the Leydig cells appeared normal. Testosterone level was significantly higher in the testes (p < 0.01) than in the plasma after 30 days of oral treatment with the standardized MeOH extract. Interestingly, eurycomanone (2) alone was detected in the rat testis homogenates by HPLC-UV and confirmed by LC/MS, and may have contributed towards the improvement of sperm quality. Thus, the plant may potentially be suitable for the management of male infertility.

Wildlife Contraception, Individuals, and Populations: How Much Fertility Control is Enough?

Author(s): Cowan, David P.; Massei, Giovanna | Abstract: The resolution of conflicts between human and wildlife interests often involves lethal control to reduce problem wildlife populations. However, lethal control has always had its limitations, the acceptable methods are becoming fewer, and public opposition is on the increase. Fertility control offers a potential alternative approach that is widely regarded as being inherently more benign. Furthermore, in some circumstances fertility control may have specific advantages over culling. The development of “single-shot” injectable immunocontraceptive vaccines that inhibit the fertility of individual animals for several years is leading to practical applications that exploit this novel technology. Further advances can be expected to lead to the emergence of a new generation of wildlife management tools. A key issue in this process is predicting what the population consequences will be for a particular species, given a specific level of induced infertility. Here, we use population modelling techniques to explore how much fertility control is enough to achieve different levels of population reduction, how long it will take to realise these reductions, and to understand how these effects are shaped by the population biology of the target species. This offers some generic conclusions, with low levels of infertility having little impact on species with high population turnover rates, while modest levels of infertility may yield useful population reductions for species with low intrinsic rates of increase, although such effects take longer to be realised in long-lived species. We also observed that there is potential for optimising the intensity of induced infertility, in terms of the proportion of breeding animals rendered infertile, and the frequency of application; so, for instance, biennial application could be more efficient than annual application for some forms of fertility control in certain species. There is increasing evidence from field studies that the survival of infertile animals is enhanced, probably because they do not incur the costs of reproduction. Our model predicts that this effect will be of limited importance for short-lived species with high intrinsic rates of increase, but it is more likely to compromise population reduction in long-lived species. We suggest that the generic modelling approach can help develop an evidence-based platform for discussing when fertility control can be regarded as a feasible, desirable, and sustainable option to manage problem wildlife.