The Case of Atypical Sexual Attractiveness in a Male Domestic Dog—A Case Study

During the ovarian cycle in domestic dogs, females do not accept males during the first days of estrus but become attractive to males from the beginning of proestrus, with this attractiveness persisting until the end of the estrus phase. It is believed that increased estradiol is responsible for the female attractiveness to the males. In this paper we describe the case of strong, but atypical attractiveness of a castrated male to various, adult, intact males, influenced by the emitted semiochemical signals. Any significant changes in the level of hormones typically involved in the process connected with estrus and responsible for sexual arousal in the males were assessed. The case animal was a 4 year old castrated male Border Collie that was extremely attractive to various males, which presented high levels of sexual arousal, with intensive sniffing and licking of the preputial area, specific vocalization, increased salivation and, finally, mating attempts. Clinical examination of the castrated male revealed a lack of testes in the scrotum and abdominal cavity confirmed by USG. Laboratory tests indicated basal levels of estradiol, testosterone, and progesterone (15.23 pg/mL, <0.05 ng/mL, 0.25 ng/mL), and sex was confirmed via cytogenetic and molecular analysis. Chemical analysis (HS-SPME) of the urine indicated a huge similarity to the profile obtained from a bitch in estrus, with an elevated level of acetophenone, which has been previously postulated in the literature as being a characteristic of the estrus phase in female domestic dogs. This case presented very atypical sexual attractiveness, particularly when taking into account the basal levels of hormones which, according to current knowledge, are responsible for the creation of attractiveness. As a hypothesis requiring verification, we propose the idea of involvement of other hormones in the creation of incidental attractiveness or increased production of compounds responsible for attractiveness (sex pheromones) resulting from metabolic events unrelated to reproductive processes. To our knowledge it is the first described case presenting this phenomenon, which, with more detailed study, could shed new light on the process of creation of sexual attraction in the domestic dog.

Urinary Metabolomics Revealed the Biological Characteristics Associated with Early Pregnancy in Pigs

Background Accurate early detection of pregnancy status is a prerequisite for effective monitoring of fertility in pig. In the early phase of pregnancy, because the embryo is small and in a free state, it is difficult to determine whether it is pregnant based on B-ultrasound examination. This calls for development of novel tools to accurately diagnose early pregnancy. Metabolomics reveal the metabolic status of cells, tissues and organisms. Results In this study, we investigated urinary metabolites in sows during early pregnancy. A total of 32 samples from 8 sows were collected at estrus and each phase of early pregnancy (days 9, 12, and 15 of gestation). Metabolites in urine samples from different sows obtained from gestation and estrus phases were analyzed via ultra-high performance liquid chromatography/mass spectrometry. A total of 530 metabolites were identified with high confidence in all samples. Compared with samples collected during the estrus phase, 269 differential metabolites were found in samples obtained during early pregnancy. Conclusions These metabolites included lipids and lipid-like molecules, organic acids and their derivatives, organic oxygen compounds, organoheterocyclic compounds, benzenoids, among others. These metabolites, such as choline and pregnanediol-3-glucuronide, play a very important role in pregnancy. They also regulate pregnancy in other animals. Our results provide novel insights into the metabolic changes in the urine of sows in the early pregnancy phase. The level of different metabolites in urine can be used to diagnosis pregnancy in sows. Understanding these metabolic changes is helpful for better management of pregnant sows.

Effect of Intrauterine Infusion of Equine Fresh Platelets-Rich Plasma (PRP) or Lyophilized PRP (L-GFequina) on Ovarian Activity and Pregnancy Rate in Repeat Breeder Purebred Arabian Mares

This study was designed to examine the effect of the intrauterine infusion of platelet-rich plasma (PRP) or equine lyophilized growth factor (L-GFequina) on the follicular growth, endometrial thickness, estrus cycle length, and pregnancy rate in purebred Arabian mares. A total of 73 purebred Arabian mares who experienced repeat breeding for three successive cycles were randomly divided into the following three groups: control group, without treatment; second Group (PRP group), in which mares were intrauterine infused with 20 mL of fresh PRP on the second day after the end of physic estrus phase; and the third group (L-GFequina Group), consisting of mares that were intrauterine infused with 20 mL of reconstituted lyophilized horse platelets growth factors (L-GFequina) on the second day after the estrus phase. The results showed no significant difference between control and treated groups in the diameter of the preovulatory follicles during the post treatment cycle. The endometrium thickness increased significantly in the L-GFequina and PRP groups ahead of the non-treated group. Intrauterine L-GFequina or PRP administration shortened the estrus cycle length. A higher pregnancy rate was recorded in the L-GFequina and PRP treated mares. In conclusion, the intrauterine infusion of L-GFequina or PRP increased the endometrial thickness and pregnancy rate and could be used to improve fertility in Arabian purebred mares who experienced from repeat breeding.

The Study of Estrous Cycle Phases Female Rats With the Use of N-Acetylcysteine in Ovarian Graft

Ovarian graft may be the target of the biochemical effects of oxidative stress caused at the time of transplantation. In order to evaluate the effect of N-acetylcysteine on the ovarian graft, regarding the estrous cycle preservation , 50 female and virgin EPM-1 Wistar rats, weighing up to 250g, originating from CEDEME of UNIFESP, were kept in adequate sanitary conditions. , receiving their own food and water. Daily vaginal smears were performed to identify the estrous phase for 8 days. The animals were randomly distributed into 05 groups: 1st Group (GTx), saline was administered subcutaneously, 2nd (NAC 150mgKg), 3rd (NAC 300mg / Kg), 4th (NAC 600mg / Kg) and 5th (NAC 1200mg / Kg) , that were administered NAC subcutaneously on the abdominal face, 60 minutes before left unilateral ovarian transplantation in retroperitoneum and contralateral oophorectomy for purposes of histomorphological analysis, with colpocytological evaluation. Euthanasia was performed by means of anesthetic lethal dose in half of the animals on the 4th postoperative day, with a single vaginal smear collection and euthanasia on the rest of the animals, between the 14th and 16th days, after the material was collected in order to define the estrus phase. It was evaluated in the graft that the animals exhibited in all groups return of estrous cycle in the later phase of the post-transplant, with better definition of regular cycle in the highest dosages of N-acetylcysteine. N-acetylcysteine induced the return of the estrous cycle in the rats’ ovarian graft , mainly in the highest dosage, proving its effectiveness in revascularization of the tissue after ischemia and reperfusion. Keywords: Acetylcysteine. Reperfusion. Histocompatibility Antigens. Menstrual Cycle. ResumoO enxerto ovariano pode ser alvo dos efeitos bioquímicos do stress oxidativo causado no momento do transplante. Com o objetivo de avaliar o efeito da N-acetilcisteína no enxerto ovariano, quanto à preservação do ciclo estral, foram utilizados 50 ratos EPM-1 Wistar, fêmeas e virgens, pesando até 250g, originários do CEDEME da UNIFESP, mantidos em adequadas condições sanitárias, recebendo ração própria e água. Realizados esfregaços vaginais diários para identificação da fase estral durante 08 dias. Os animais foram distribuídos aleatoriamente em 05 grupos: 1º Grupo (GTx), administrada solução salina via subcutânea, 2º (NAC 150mgKg), 3º (NAC 300mg/Kg), 4º (NAC 600mg/Kg) e 5º (NAC 1200mg/Kg), aos quais foi administrada NAC por via subcutânea em face abdominal, 60 minutos antes do transplante unilateral esquerdo do ovário em retroperitônio e à ooforectomia contra-lateral para fins de análise histomorfológica, com avaliação colpocitológica. A eutanásia foi realizada por meio da dose letal do anestésico em metade dos animais no 4º dia de pós-operatório, realizado única coleta de esfregaço vaginal e a eutanásia no restante dos animais, entre o 14 º e 16º dia, após a coleta do material para definição da fase estro. Foi avaliado no enxerto que os animais apresentaram em todos os grupos retorno de ciclo estral na fase mais tardia do pós-transplante, com melhor definição de ciclo regular nas dosagens mais elevadas de N-acetilcisteína. A N-acetilcisteína induziu o retorno do ciclo estral no enxerto ovariano de ratas, principalmente na maior dosagem comprovando sua eficácia na revascularização do tecido após isquemia e reperfusão. Palavras-chave: Acetilcisteína. Reperfusão. Transplante. Ciclo Ovariano.

Analysis of Expression Profiles of mRNA and lncRNA in Oviduct During Estrus Phase of Sheep (Ovis Aries) with Two Fecundity (FecB Gene) Genotypes

Background:In sheep, FecB is the essential biomarker of the fertility, previous researches have provided a detailed insight on the regulation involved estrus phase and FecB in the reproductive-related tissues including hypothalamus, pituitary, and ovary. However, as the host of embryo development and connection between the ovary and the uterus, little is known about the interaction between mRNAs and lncRNAs in sheep oviduct. In the present study, RNA-Seq was performed to identify the transcriptomic profiles of mRNAs and lncRNAs in oviduct during estrus phase of sheep with FecBBB/++ genotypes.Results:In total, 21,863 lncRNAs and 43,674 mRNAs were identified, 57 DE lncRNAs and 637 DE mRNAs were revealed in the comparisons between follicular phase and luteal phase, 26 DE lncRNAs and 421 DE lncRNAs were revealed in the comparisons between FecB BB genotype and FecB ++ genotype. Functional enrichment analysis suggested that GO and KEGG terms related to reproduction such as SAGA complex, ATP-binding cassette (ABC), Nestin, and Hippo signalling pathway. DE-interaction network suggested that LNC_018420 maybe the key regulators related to embryo development in sheep oviduct.Conclusion:This was the first study to reveal the transcriptomic profiles of mRNAs and lncRNAs in the oviduct of FecB BB/++ sheep at estrus phase using RNA-Seq. Our findings can provide new understanding on the molecular mechanisms of mRNAs and lncRNAs underlying sheep embryo development and also opening new lines of investigation in sheep reproduction.

Relationship between the value of the estrous detector measurement result and serum progesterone level in Etawa crossbred goats after estrous synchronization

This study was conducted to determine the relationship between the estrous detector's reading value (Draminski®, Dramiński S.A, Poland) and serum progesterone levels on Etawa crossbred does. Eighteen healthynon-prehnant Etawa crossbreds does weighing 20-25 kg, in their first parity were used in this study. The does were divided equally into two groups. In the two groups, the does were synchronized their estrus by means of intramuscular and intravulvar injection of 7.5 mg prostaglandin Fɑ (PGF2ɑ) twice in 11-day intervals. After the second injection, the estrous detector value measurement, visual observation of the estrus sign, and blood sampling were conducted for five days. The blood sample was processed into the serum to examine for progesterone levels by ELISA. The data analysis was using the simple correlation method on SPSS type 23 for windows. The result showed that a positive relationship (p <0.05) with coefficient correlation (r) was 0.609 between the values of the estrous detector measurement with serum progesterone levels. It could be concluded that the estrous detector's low value indicates that the Etawa crossbred was in the estrus phase and followed by the decreasing of progesterone level but not reaching the basal level.

Localization (and profiles) of tyrosinephosphorylated proteins in female reproductive organs of adult rats

Objective: Tyrosine phosphorylation is an essential process in many biological systems, including the male reproductive system. The presence of tyrosine-phosphorylated (TyrPho) proteins has been well documented in male reproductive organs, but research in fertile females is still limited.Methods: The ovary, oviduct, and uterus of adult female Sprague-Dawley rats in the estrus phase were used to localize TyrPho proteins using an immunohistochemical technique. These proteins were separated and their expression patterns were examined by sodium dodecyl sulfatepolyacrylamide gel electrophoresis and Western blot analysis, respectively.Results: TyrPho proteins were localized in the cytoplasm of the oocyte except the antral fluid; in the granulosa cells, theca cells, and stromal cells of the ovary; at the apical surface of oviductal epithelial cells; and in the basal epithelium and submucosa of the uterine wall. Moreover, we found that 72-, 43-, and 28-kDa TyrPho proteins were localized in the ovary, while 170-, 55-, and 43-kDa proteins were localized in the oviduct. In the uterus, we detected four major bands, corresponding to 61-, 55-, 54-, and 43-kDa TyrPho proteins.Conclusion: Given that these TyrPho proteins were found in major reproductive organs in the estrus phase, these proteins may play important roles in female fertility.

Estrus Synchronization Using Prostaglandin F2α (PGF2α) and Combination of PGF2α and Gonadotropin-Releasing Hormone In Ongole Crossbred

The response of estrus can be enhanced by providing PGF2α and a combination of PGF2αGnRH so that the time of mating using Artificial Insemination (AI) can be held precisely. This study aims to compare the response of estrus synchronization using PGF2α compare to PGF2α-GnRH. Twenty female cattle Ongole Crossbreds, 4-6 years, BCS average of 3-4 were used in this research. Cattle were selected based on their luteal phase at the days of 7 to 18 by rectal examination. Then, all of the cattle were injected by cloprostenol® with a dose of 500 ug intrauterine. Ten of the animals were injected by 2.5 mL of Fertagyl® intramuscularly after 48 hours then continued observing and response estrus detection. The onset of estrus using PGF2α was detected on 75.94 ± 0.78, while 41.57 ± 28.40 hours for PGF2α-GnRH Using statistical, there was a significant difference (P<0.05). Duration of estrus phase using PGF2α was 24.65 ± 0.49 whereas PGF2α-GnRH was 10.88 ± 7.45 hours. Using statistical methods, there was a significant difference (P <0.05). It could be concluded that the use of PGF2α was more effective for the duration and intensity of estrus, and a combination of PGF2α -GnRH is more effective for stimulating estrus.

Comprehensive Analysis of mRNAs and miRNAs in the Ovarian Follicles of Uniparous and Multiple Goats at Estrus Phase

Background Fertility is an important economic trait in production of meat goat, and follicular development plays an important role in fertility. Despite many mRNAs and microRNAs (miRNAs) have been found in playing critical roles in ovarian biological processes, the interactions between mRNAs and miRNAs in follicular development is not yet completely understood. In addition, less attention has been given to the single follicle (dominant or atretic follicle) in goat. The study was aimed to identify mRNAs, miRNAs and signaling pathways as well as their interaction networks in the ovarian follicles (large follicles and small follicles) of multiple and uniparous goats (Chuanzhong Black Goats) at estrus phase by using a deep RNA-sequencing (RNA-seq) method.Results The result showed that there were more large follicles in multiple than in uniparous goats ( P <0.05), while no difference was observed in small follicles between them ( P >0.05). For the small follicles of multiple and uniparous goats at estrus phase, 289 differentially expressed mRNAs (DEmRNAs) and 16 DEmiRNAs were identified; and for the large follicles, 195 DEmRNAs and 7 DEmiRNAs were identified. Ovarian steroidogenesis and steroid hormone biosynthesis were significantly enriched in small follicles, while ABC transporters and steroid hormone biosynthesis in large follicles. The results of qRT-PCR were generally consistent with the RNA-seq data. The mRNA-miRNA interaction network showed that CD36 (miR-122, miR-200a, miR-141), TNFAIP6 (miR-141, miR-200a, miR-182), CYP11A1 (miR-122), SERPINA5 (miR-1, miR-206, miR-133a-3p, miR-133b) and PTGFR (miR-182, miR-122) might be related to fertility, but need further verification.Conclusion This study provides the first identification of the DEmRNAs and DEmiRNAs as well as their interactions in the follicles of multiple and uniparous goats at estrus phase by using RNA-seq technology. These analyses provide new clues to uncover molecular mechanisms and signaling networks of goat reproduction which could be potentially used to increase ovulation rate and kidding rate in goat.