Local production of 17ß-oestradiol in the endometrium during the implantation window: a pilot study.

Abstract: Sex steroids are converted to bioactive metabolites and vice versa by endometrial steroid-metabolising enzymes. Studies indicate that alterations in this metabolism might affect endometrial receptivity. This pilot study determined whether the endometrial formation and inactivation of 17ß-oestradiol differed between the supposedly embryo-receptive endometrium and non-receptive endometrium of women undergoing IVF/intracytoplasmic sperm injection (ICSI). Endometrial biopsies were obtained from IVF/ICSI patients 5-8 days after ovulation in a natural cycle, prior to their second IVF/ICSI cycle with fresh embryo transfer (ET). Endometrial biopsies from patients who achieved clinical pregnancy after fresh ET (n = 15) were compared with endometrial biopsies from patients that did not conceive after fresh ET (n = 15). Formation of 17ß-oestradiol (oxidative 17ß-hydroxysteroid dehydrogenases (HSDs)), oestrone (reductive HSD17Bs) and inhibition of HSD17B1 activity were determined by high-performance liquid chromatography. The endometrial transcriptome was profiled using RNA sequencing followed by principal component analysis and differentially expressed gene analysis. The false discovery rate-adjusted P < 0.05 and log fold change >0.5 were selected as the screening threshold. Formation and inactivation of 17ß-oestradiol resulted similar between groups. Inhibition of HSD17B1 activity was significantly higher in the non-pregnant group when only primary infertile women (n = 12) were considered (27.1%, n = 5 vs 16.2%, n = 7, P = 0.04). Gene expression analysis confirmed the presence of HSD17B1 (encoding HSD17B1), HSD17B2 (encoding HSD17B2) and 33 of 46 analysed steroid metabolising enzymes in the endometrium. In the primary infertile subgroup (n = 10) 12 DEGs were found including LINC02349 which has been linked to implantation. However, the exact relationship between steroid-metabolising enzyme activity, expression and implantation outcome requires further investigation in larger, well-defined patient groups. Lay summary: Sex hormones are produced and broken down by enzymes that can be found in the endometrium (the inner lining of the womb). This enzyme activity might influence the chances of becoming pregnant. We compared (i) enzyme activity in the endometrium of 15 women who did and 15 women who did not become pregnant in their second in vitro fertilisation attempt, (ii) how enzyme activity can be blocked by an inhibitor, and (iii) differences in gene expression (the process by which instructions in our DNA are converted into a product). Enzyme activity was similar between groups. We found that in women who have never been pregnant in the past, inhibition of enzyme activity was higher and found differences in a gene that has been linked to the implantation of the embryo, but future studies should be performed in larger, well-defined patient groups to confirm these findings.

Comparative studies on ovary and serum hormone levels in silver and blue foxes and their hybrids.

The silver and blue fox appear closely related despite their belonging to different genera, and the male and female fox hybrids are completely sterile in mating inter se or back to either of the parental groups. In the present study, we compared ovarian morphology and serum hormone concentrations among silver foxes, blue foxes, HSBs (silver fox male × blue fox female hybrids) and HBSs (blue fox male × silver fox female hybrids) during the pre-breeding and the breeding period. We found that either the fertile blue and silver foxes or the infertile hybrid foxes showed typical characteristics of ovary, with primordial, primary, secondary, antral and Graafian follicles. The diameters and quantities of different follicle classes in fertile foxes were greater than that in hybrids during the two period, while the thickness of zona pellucida (ZP) represented an opposite trend (P < 0.05). The relative weight of bilateral ovaries in fertile foxes was greater than that in hybrids during the pre-breeding period, while mean serum FSH represented an opposite trend during the breeding period (P < 0.05). The mean serum estradiol in fertile foxes was greater than that in HBS foxes during the pre-breeding period, and greater than hybrids during the breeding period (P < 0.05). These findings indicated that in the infertile hybrid foxes, lower estradiol levels produced by lesser and smaller antral follicles, thicker ZP during the two period, higher FSH levels during the breeding period, could be the basis of interpretations in cases of female fox hybrid infertility.

Maternal Kdm2a-mediated PI3K/Akt signaling and E-cadherin stimulate the morula-to-blastocyst transition revealing crucial roles in early embryonic development.

Histone methylation plays an essential role in oocyte growth and preimplantation embryonic development. The modification relies on histone methyl-transferases and demethylases, and one of these, lysine-specific demethylase 2a (Kdm2a), is responsible for modulating histone methylation during oocyte and early embryonic development. The mechanism of how Kdm2a deficiency disrupts early embryonic development and fertility remains elusive. To determine if maternally deposited Kdm2a is required for preimplantation embryonic development, the expression profile of Kdm2a during early embryos was detected via immunofluorescence staining and RT-qPCR. The Kdm2a gene in oocytes was specifically deleted with the Zp3-Cre/LoxP system and the effects of maternal Kdm2a loss were studied through a comprehensive range of female reproductive parameters including fertilization, embryo development, and the number of births. RNA transcriptome sequencing was performed to determine differential mRNA expression, and the interaction between Kdm2a and the PI3K/Akt pathway was studied with a specific inhibitor and activator. Our results revealed that Kdm2a was continuously expressed in preimplantation embryos and loss of maternal Kdm2a suppressed the morula-to-blastocyst transition, which may have been responsible for female subfertility. After the deletion of Kdm2a, the global H3K36me2 methylation in mutant embryos was markedly increased, but the expression of E-cadherin decreased significantly in morula embryos compared to controls. Mechanistically, RNA-seq analysis revealed that deficiency of maternal Kdm2a altered the mRNA expression profile, especially in the PI3K/Akt signaling pathway. Interestingly, the addition of a PI3K/Akt inhibitor (LY294002) to the culture medium blocked embryo development at the stage of morula; however, the developmental block caused by maternal Kdm2a loss was partially rescued with a PI3K/Akt activator (SC79). In summary, our results indicate that loss of Kdm2a influences the transcriptome profile and disrupts the PI3K/Akt signaling pathway during the development of preimplantation embryo. This can result in embryo block at the morula stage and female subfertility, which suggests that maternal Kdm2a is a potential partial redundancy with other genes encoding enzymes in the dynamics of early embryonic development. Our results provide further insight into the role of histone modification, especially on Kdm2a, in preimplantation embryonic development in mice.

First detection and phylogenetic analysis of porcine circovirus 3 in female donkeys with reproductive disorders.

BACKGROUND: PCV3 is a pathogen associated with porcine dermatitis and nephropathy syndrome (PDNS)-like clinical signs, reproductive failure, and cardiac and multiorgan inflammation, which was newly identified in 2016 in sows in USA. Recently, PCV3 has also been identified from several non-porcine species like (cattle, dog, wild boar, deer, mice and ticks). However, PCV3 infection in donkey is not well established. Since 2019, 300 blood samples were collected from female donkey, which was characterized by abortion and sterility, in Liaocheng city of China. RESULTS: In the present study, an investigation of PCV3 in donkey blood samples was undertaken employing by real time PCR. Positive rates of PCV3 in donkeys reach to 21.0 %. In addition, one full-length PCV3 genome sequence was obtained, and it had a highest identity with porcine circovirus 3 PCV3/CN/Nanjing2017 strain and is clustered to PCV3a genotype based on ORF2 sequences. CONCLUSIONS: This is the first report of detection of PCV3 from female donkeys presenting reproductive failure in large-scale donkey farms, China. In addition, the PCV3 strain identified in this study shared the closest relationship with those from porcine, suggesting that PCV3 may be transmitted from pigs to donkeys. Totally, PCV3 infection in donkey should be concerned although the association between it and reproductive failure are not better understood.

Use of Hysteroscopy for Diagnosing Causes of Infertility in Camelids.

Hysteroscopy in alpacas and llamas allows for the identification of abnormalities on the surface or within the endometrium that cannot be identified with other methods. Hysteroscopy also allows for site-directed endometrial cytology, culture, and biopsy to achieve a definitive diagnosis. Even when no cause for infertility can be found, previously infertile females tend to become pregnant and maintain their pregnancies to term following the hysteroscopic procedure. This therapeutic effect may be a response to pre-hysteroscopy estrogen treatment, dilation of the uterine horns during hysteroscopy, and/or posttreatment uterine lavages. Complications following hysteroscopy have not been reported in camelids.

Characterization of immunological, biochemical and inflammatory response of clinical and subclinical endometritis in ewes in the subtropics.

Pluriparus Ossimi (n = 50) ewes were used to investigate the immune profile of the affected ewes to accurately diagnose clinical and subclinical endometritis and associations with biochemical variables. Ewes were slaughtered and animals were classified into control (no fertility problems), subclinical endometritis (SCE) and clinical endometritis (CE) groups based on pre-slaughter determinations of conception failure. Serum was collected from ewes to estimate concentrations of pro-inflammatory cytokines including interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and interleukin-8 (IL-8) as well as nitric oxide (NO) concentration. The results from immunological evaluations indicated there were greater (P < 0.001) serum concentrations of IL-6, IL-8, TNF-α and NO in ewes classified with SCE and CE as compared to ewes of the control group. Furthermore, values for concentrations of TNF-α were positively correlated with IL-6 and IL-8 concentrations in ewes of the SCE and CE groups. In ewes classified with CE and SCE there were greater (P < 0.01) concentrations of blood glucose, ALT, AST, urea and creatinine than in ewes of the control group. It is concluded that serum pro-inflammatory cytokines IL-6, IL-8, and TNF-α are diagnostic markers for CE and SCE in ewes and serve as a criterion for different inflammatory complications in ewes classified as having CE or SCE.

The Freemartin Cattle and Clinical Transplantation: From the Ancients to Modern Day.

This historical retrospective explores the study of the freemartin condition and its impact on the discovery of immunologic tolerance and the field of transplant surgery-from the ancient Romans, to early modern anatomists Valsalva, Scarpa, and Hunter, to contemporary immunologists Owen, Medawar, and Billingham, and to legendary transplant surgeon Joseph Murray. The legacy of freemartin cattle in the understanding of acquired tolerance and transplant immunology represents generations of scientific inquiry guided by careful observation and occasional serendipity, and the present-day immunologists and surgeons exploring immune transplant tolerance owe much to the history of the freemartin, several millennia in the making.

Genital abnormalities associated to lack of uterine adenogenesis or endometrial gland dysgenesis of female dromedary camels (Camelus dromedarius).

Background: The developmental disruption of the müllerian duct and the endometrial dynamic can generate genital lesions that could contribute to infertility. Aim: This paper discusses two cases of genital conditions associated to endometrial gland pathologies in nulliparous female camels. Methods: Macroscopic examinations and histopathological description were performed on congenital and acquired genital abnormalities with endometrial gland anomalies. Results: The first case is endometrial gland agenesis associated to unilateral uterine aplasia, and the second case is endometrial gland dysgenesis associated to metritis. The prevalence of each case is estimated to be 0.6%. The most specific microscopic features associated to the endometrial gland agenesis were the presence of endometrial stromal proliferation and homogenous hyalinization of the myometrium. The acute metritis was associated to endometrial-activated stroma with focal infiltration with inflammatory cells on the endometrium and myometrium and the spontaneous endometrial gland dysgenesis. Conclusion: This study reveals the importance of congenital abnormalities during the routine reproductive examination of peripubertal animals, as well as the association of histopathological complementary examination for the research functional and inflammatory anomalies of the uterus. Genetic screening of breeders would be very important in the search for genetic risk factors associated with these congenital pathologies, which can be disseminated by reproductive biotechnologies.

Weighted Gene Correlation Network Meta-Analysis Reveals Functional Candidate Genes Associated with High- and Sub-Fertile Reproductive Performance in Beef Cattle.

Improved reproductive efficiency could lead to economic benefits for the beef industry, once the intensive selection pressure has led to a decreased fertility. However, several factors limit our understanding of fertility traits, including genetic differences between populations and statistical limitations. In the present study, the RNA-sequencing data from uterine samples of high-fertile (HF) and sub-fertile (SF) animals was integrated using co-expression network meta-analysis, weighted gene correlation network analysis, identification of upstream regulators, variant calling, and network topology approaches. Using this pipeline, top hub-genes harboring fixed variants (HF × SF) were identified in differentially co-expressed gene modules (DcoExp). The functional prioritization analysis identified the genes with highest potential to be key-regulators of the DcoExp modules between HF and SF animals. Consequently, 32 functional candidate genes (10 upstream regulators and 22 top hub-genes of DcoExp modules) were identified. These genes were associated with the regulation of relevant biological processes for fertility, such as embryonic development, germ cell proliferation, and ovarian hormone regulation. Additionally, 100 candidate variants (single nucleotide polymorphisms (SNPs) and insertions and deletions (INDELs)) were identified within those genes. In the long-term, the results obtained here may help to reduce the frequency of subfertility in beef herds, reducing the associated economic losses caused by this condition.

Inflammatory response in repeat breeder buffaloes.

The aim of this study was to investigate the repeat breeding condition in Italian Mediterranean buffaloes that failed to conceive after at least 300 days in milk. The trial was carried out on 40 pluriparous Italian Mediterranean buffaloes with more than 300 days in milk. All the animals underwent ultrasound examination to assess endometrial thickness and oestrus synchronization by the Ovsynch-TAI Program. On the day of oestrus, blood samples were collected for the haemocytometric cell count and biochemical assay, and the animals were slaughtered in a local abattoir. A post-mortem uterine flushing was performed using sterile saline for microbiological analyses. Furthermore, uterine biopsies were carried out for histopathological assessment. Finally, endometrial samples were used for real-time PCR (RT-PCR) analysis to evaluate the expression of genes involved in innate immune recognition of pathogens and the inflammatory response, such as Toll-like receptor (TLR)1, TLR8, interleukin (IL)-1ß, IL-6, IL-8, COL4A2, connective tissue growth factor (CTGF), and cysteine-rich angiogenic inducer 61 (CYR61). Statistical analysis was performed by one-way ANOVA. Based on the infiltration of lymphocytes and plasma cells or endometrial gland, lymphatic, and blood vessel ectasia recorded in the histopathological examination, the animals were classified into three groups: healthy (H Group; n = 5), moderate endometritis (M Group; n = 25), and severe endometritis (S Group; n = 10). A significantly greater (P < 0.01) endometrial thickness was recorded in the S Group compared to that in the H and M Group (1.07 ± 0.03 vs. 0.70 ± 0.07 and 0.81 ± 0.04 cm in the S, H, and M Group, respectively). The white blood cell count was lower in the H Group compared to that in the M and S Group (6.3 ± 0.6 vs. 9.3 ± 0.4 and 10.5 ± 0.5 in the H, M, and S Group, respectively). To perform RT-PCR analysis, five animals from groups M and S were randomly selected in order to have balanced results. A higher (P < 0.01) expression of TLR1, together with a lower expression of COL4A2, IL-1ß, IL-6, IL-8, and CYR61, was recorded in the H Group, compared to both the M and S Groups. In conclusion, about 90% of repeat breeder buffaloes show moderate or severe endometritis, associated with an altered histopathological endometrial profile and altered mRNA expression of pro-inflammatory and fibrotic factors.

Analysis of the uterine lumen in fertility-classified heifers: II. Proteins and metabolites†.

Survival and growth of the bovine conceptus is dependent on endometrial secretions or histotroph. Previously, serial blastocyst transfer was used to classify heifers as high fertile (HF), subfertile (SF), or infertile (IF). Here, we investigated specific histotroph components (proteins and metabolites) in the uterine lumen of day 17 fertility-classified heifers. Interferon tau (IFNT) was more abundant in uterine lumenal fluid (ULF) of pregnant HF than SF animals as the conceptus was longer in HF heifers. However, no differences in endometrial expression of selected classical and nonclassical interferon-stimulated genes (ISGs) were observed, suggesting that IFNT signaling in the endometrium of pregnant HF and SF heifers was similar. Pregnancy significantly increased the abundance of several proteins in ULF. Based on functional annotation, the abundance of a number of proteins involved in energy metabolism, oxidative stress, amino acid metabolism, and cell proliferation and differentiation were greater in the ULF of pregnant HF than SF heifers. Metabolomics analysis found that pregnancy only changed the metabolome composition of ULF from HF heifers. The majority of the metabolites that increased in the ULF of pregnant HF as compared to SF heifers were associated with energy and amino acid metabolism. The observed differences in ULF proteome and metabolome are hypothesized to influence uterine receptivity with consequences on conceptus development and survival in fertility-classified heifers.

Premature centromere division (PCD) identified in a hucul mare with reproductive difficulties.

A hucul mare with reproductive abnormalities was examined during karyotype analysis. The karyotype was analysed based on evaluation of 860 metaphase plates in chromosome preparations. The use of fluorescence in situ hybridization (FISH) with an X chromosome painting probe showed premature X chromosome separation in 9.5% cases of examined chromosome spreads. In this report, we present the first identify premature centromere division (PCD) as a possible cause of abnormal X chromosome morphology in horses and as a probable cause of reproductive difficulties.

Pregnancy associated glycoproteins (PAGs) and pregnancy loss in high vs sub fertility heifers.

Reproductive inefficiency and infertility are major financial burdens to domestic livestock. Variables associated with these reproductive losses during early gestation include contributions from the oocyte, uterus, sperm, embryo and placenta. Bovine pregnancy associated glycoproteins (PAG) are produced by the binucleate cells of the ruminant placenta and can be used to diagnose pregnancy. Increased circulating concentrations of PAG early in gestation have been correlated with pregnancy success and decreased concentrations are predictive of impending embryonic mortality in both beef and dairy cattle. The objectives of the current study were to determine whether: 1) heifer fertility status is associated with circulating concentrations of PAG and pregnancy loss; and 2) PAG concentrations within the same animal are repeatable across multiple pregnancies. We hypothesized maternal PAG concentrations would be increased in high fertility compared to subfertile heifers but not repeatable across subsequent pregnancies in the same heifer. Serial embryo transfer (ET; n = 4 rounds) was used to classify predominately Angus heifers (n = 92) as highly fertile (HF = 30; 100% pregnancy success) or subfertile (SF = 62; average = 33%; range = 25-75% pregnancy success) based on day 28 ultrasound diagnosis. Blood samples were collected at both day 28 and 44 for quantification of circulating PAG concentrations by an in house PAG ELISA with antibodies raised against early secreted PAGs. Pregnancy was terminated at day 44 of gestation and heifers were allowed 30 days recovery before synchronization for the next ET. Only heifers that were diagnosed pregnant by ultrasound were used in this study (HF: n = 30, SF: n = 62). Serum concentrations of PAGs were not different between HF (5.90 ±â€¯0.27 ng/mL) and SF (5.56 ±â€¯0.31 ng/mL; P = 0.16) heifers at day 28 of gestation nor was there a difference at day 44 of gestation (P = 0.32). Subfertile heifers had increased pregnancy loss between days 28 and 44 of gestation. Based on odds ratio analysis, SF heifers had a 2.41 times chance to undergo pregnancy loss between day 28-44 compared to HF heifers (P < 0.05). There was no correlation (P > 0.05) in maternal circulating concentrations of PAG between pregnancies on day 28 or 44 of gestation in samples obtained from HF heifers. In summary, circulating concentrations of PAG are not different between HF and SF heifers; however, HF classified heifers have decreased pregnancy loss between days 28 and 44 of gestation.

Successful Implantation and Live Birth Following Autologous Platelet-rich Plasma Treatment for a Patient with Recurrent Implantation Failure and Chronic Endometritis.

BACKGROUND/AIM: Patients diagnosed with chronic endometritis (CE) may fail to respond to standard antibiotic treatment. The driver behind the approach reported here was the imperative need for alternative therapeutic solutions. CASE REPORT: This case report presents a woman with CE and premature ovarian insufficiency having experienced repeated implantation failures following donated embryo transfers. The patient was diagnosed with CE through hysteroscopy, microbiological analysis and scanning electron microscopy. Following the suggested antibiotic treatment, she underwent a new embryo transfer, but with subsequent pregnancy loss. Following a second antibiotic scheme, all diagnostic procedures certified the persistence of CE. The patient underwent autologous, intrauterine platelet-rich plasma treatment and a subsequent embryo transfer. The diagnostic procedures indicated no signs of CE, while the embryo transfer resulted in a twin pregnancy and birth. CONCLUSION: Platelet-rich plasma may be employed as a first-line CE treatment, especially for patients who fail to respond to conventional antibiotic schemes.

Symposium review: Mechanisms of disruption of fertility by infectious diseases of the reproductive tract.

Diseases of postpartum dairy cows impair reproductive processes, resulting in prolonged anestrus, reduced conception, and increased pregnancy attrition, regardless of whether the initial disease precedes insemination (even by many weeks), occurs close to insemination, or follows fertilization. Bacteria and their products activate pattern recognition receptors that respond to pathogen-associated molecular patterns (PAMP). These receptors include toll-like receptors (TLR), nucleotide-binding oligomerization domain (NOD)-like receptors and others, and their activation culminates in upregulation of proinflammatory cytokines such as IL-1ß, IL-18, and tumor necrosis factor-α. These may have direct effects on the uterus and conceptus. Importantly, however, these inflammatory mediators, as well as the bacterial products, make their way to the ovary via the general circulation (even from distant sites) or possibly by using the countercurrent vascular mechanism that normally transports endometrial prostaglandin to the ipsilateral ovary. Endotoxin reaches concentrations in follicular fluid that exceed those found in the circulation or even in the infected uterus. Ovarian follicular cells also express TLR and can respond directly to bacterial products including endotoxin, impairing their function. Inflammation is accompanied by increased oxidative stress. The process of oocyte development from activation of primordial oocytes to potential ovulation spans 4 mo. Competence by an oocyte encompasses the ability to undergo not only fertilization but also a complex cytoplasmic maturation that lays the foundation for completion of meiosis at the appropriate time, the transition to mitosis in the zygote, and further development of the conceptus. Oocyte maturation relies on intimate association between cumulus cells and the oocyte, characterized by gap junctions through which molecules of various sizes pass. Signaling also occurs in the oocyte-to-cumulus cell direction. Because both granulosa and theca interna cells are capable of responding to inflammatory mediators, with observed alterations in some functions, it seems likely that disturbed ovarian follicular function may contribute to failure of oocytes to become fully competent, even if the insult occurs well before ovulation. Therefore, interruption of normal fertility by uterine infections may be mediated at the level of the uterine environment but the effect on the ovary and oocyte is likely to be even more important.

Reproductive disorders in does experimentally infected by Toxoplasma gondii / Distúrbios reprodutivos em cabras experimentalmente infectadas por Toxoplasma gondii

The aim of this study was to describe the reproductive disorders related to experimental infection by artificial insemination with semen contaminated with Toxoplasma gondii of four goats in the chronic phase of the infection. In the end of the study, the does were submitted to necropsy, and PCR and histopathological evaluations were performed. Among infected does that exhibited embryonic loss, two were in anestrus and two exhibited repeated estrus. One of the latter animals exhibited clinical signs of estrus at seven-day intervals, whereas the other had a 21-day estrous cycle. However, both does were naturally mated on subsequent natural estrous and were not able to get pregnant until the end of the experiment (90 d). Two of the goats exhibited abnormalities in the ultrasound examinations, one of which was an ovarian cyst, while the other was a hydrosalpinx, both of which were confirmed in the post-mortem examination. The main microscopic injuries in this group were neutrophilic infiltration of the lungs, interstitial glomerulonephritis and neutrophilic infiltration of the liver. T. gondii DNA was found in the organs (heart and brain) of three does. In conclusion, does infected with Toxoplasma gondii in semen at the time of artificial insemination display reproductive disorders in the chronic phase of infection that might be associated with toxoplasmosis.

Objetivou-se descrever os distúrbios reprodutivos associados à infecção experimental por Toxoplasma gondii através da inseminação artificial com sêmen contaminado em quatro cabras no estágio crônico da infecção. As características do trato reprodutor foram avaliadas através de ultrassonografia transretal, visando o diagnóstico gestacional ou de desordens reprodutivas, após a infecção experimental. Ao final do experimento, os animais foram necropsiados e avaliações histopatológicas e PCR foram realizados. Dentre os animais infectados que exibiram mortalidade embrionária, duas apresentaram anestro e duas apresentaram repetição de estro, sendo que destas uma apresentou intervalos entre estros reduzido (sete dias) e outra em intervalo regular (21 dias). Todavia, ambas foram submetidas a monta natural durante os estros naturais subsequentese não foi confirmada gestação até o final do experimento (90 dias). Duas cabras exibiram alterações nos exames de ultrassonografia, sendo identificadas um cisto ovariano, e uma hidrossalpinge, ambas confirmadas no exame post-mortem. As principais lesões microscópicas nesse grupo foram infiltração neutrofílica dos pulmões, glomerulonefrite intersticial e infiltração neutrofílica do fígado. O DNA de T. gondii foi encontrado nos órgãos (coração e cérebro) de três cabras. Em conclusão, cabras infectadas comsêmen contendoT. gondii no momento da inseminação artificial apresentam distúrbios reprodutivos na fase crônica da infecção que podem estar associados à toxoplasmose.

Impairment of the antifibrotic prostaglandin E2 pathway may influence neutrophil extracellular traps-induced fibrosis in the mare endometrium.

Prostaglandin E2 (PGE2) has contradictory effects in many organs. It may have proinflammatory, anti-inflammatory, or anti-fibrotic roles, depending on the type of receptors to which it binds. By signaling through its receptors EP2 and EP4, PGE2 mediates anti-inflammatory and anti-fibrotic actions. In spite of chronic endometrial fibrosis (endometrosis) being a major cause of mare infertility, its pathogenesis is not fully understood. We have shown that contact of mare endometrium in vitro with neutrophil extracellular traps (NETs) proteases favors endometrial collagen type I production. Therefore, we investigated the involvement of the PGE2 pathway in collagen deposition in mare endometrium, challenged in vitro with proteases present in NETs. Mare endometria (Kenney and Doig categories I/IIA and IIB/III), obtained in the follicular phase (FLP) and mid-luteal phase (MLP), were incubated for 24 h with components found in NETs (elastase, cathepsin-G, and myeloperoxidase). Secretion of PGE2 and transcripts for specific PGE synthase (PGES) and PGE2 receptors (EP2 and EP4) were evaluated. Impaired PGE2 production and low EP2 transcript abundance depended on the endometrial category and estrous cycle phase. Impairment of PGE2 and/or EP2 might play a role in FLP (category IIB/III) and MLP (I/IIA) endometrial fibrogenesis because of the reduction in its antifibrotic capacity. In conclusion, priming of the endometrium with endogenous ovarian steroids might inhibit the antifibrotic PGE2 pathway either in healthy or pathologic tissues with collagen formation after NETs proteases action.

Clinical application of hysteroscopic hydrotubation for unexplained infertility in the mare.

BACKGROUND: Therapeutic techniques for oviductal obstruction in the mare are limited. Nonsurgical and retrograde flushing may be an attractive alternative to current treatment methods for oviductal blockage. OBJECTIVES: To evaluate hysteroscopic selective hydrotubation as a treatment option for presumptive equine oviductal blockage. STUDY DESIGN: Retrospective case series. METHODS: A quantity of 10 mL of saline was flushed through the oviducts in 28 standing sedated mares, which had reproductive histories of unexplained subfertility, by inserting a catheter into the uterotubal junction under endoscopic guidance. All mares in the study had been mated through several cycles (2-20 oestrous cycles) by known fertile stallions prior to treatment, with no evidence of conception. The average number of cycles for each mare prior to treatment was 6.5 ± 4.5. RESULTS: Saline was successfully infused into a total of 50 oviducts. Of 28 mares, 26 conceived after the treatment. The average number of cycles for each mare to become pregnant after treatment was 1.8 ± 0.8. MAIN LIMITATIONS: Diagnosis of blocked oviducts was presumptive, and pretreatment infertility was used as the control. CONCLUSIONS: This study revealed that hysteroscopic hydrotubation using saline improved pregnancy rates in mares in which oviductal blockage was suspected as a cause of unexplained subfertility.

Maternal variant in the upstream of FOXP3 gene on the X chromosome is associated with recurrent infertility in Japanese Black cattle.

BACKGROUND: Repeat breeding, which is defined as cattle failure to conceive after three or more inseminations in the absence of clinical abnormalities, is a substantial problem in cattle breeding. To identify maternal genetic variants of repeat breeding in Japanese Black cattle, we selected 29 repeat-breeding heifers that failed to conceive following embryo transfer (ET) and conducted a genome-wide association study (GWAS) using the traits. RESULTS: We found that a single-nucleotide polymorphism (SNP; g.92,377,635A > G) in the upstream region of the FOXP3 gene on the X chromosome was highly associated with repeat breeding and failure to conceive following ET (P = 1.51 × 10-14). FOXP3 is a master gene for differentiation of regulatory T (Treg) cells that function in pregnancy maintenance. Reporter assay results revealed that the activity of the FOXP3 promoter was lower in reporter constructs with the risk-allele than in those with the non-risk-allele by approximately 0.68 fold. These findings suggest that the variant in the upstream region of FOXP3 with the risk-allele decreased FOXP3 transcription, which in turn, could reduce the number of maternal Treg cells and lead to infertility. The frequency of the risk-allele in repeat-breeding heifers is more than that in cows, suggesting that the risk-allele could be associated with infertility in repeat-breeding heifers. CONCLUSIONS: This GWAS identified a maternal variant in the upstream region of FOXP3 that was associated with infertility in repeat-breeding Japanese Black cattle that failed to conceive using ET. The variant affected the level of FOXP3 mRNA expression. Thus, the results suggest that the risk-allele could serve as a useful marker to reduce and eliminate animals with inferior fertility in Japanese Black cattle.