Metagenomic analysis unravels novel taxonomic differences in the uterine microbiome between healthy mares and mares with endometritis.

BACKGROUND: The application of high throughput technologies has enabled unravelling of unique differences between healthy mares and mares with endometritis at transcriptomic and proteomic levels. However, differences in the uterine microbiome are yet to be investigated. OBJECTIVES: The present study was aimed at evaluating the differences in uterine microbiome between healthy mares and mares with endometritis. METHODS: Low-volume lavage (LVL) samples were collected from the uterus of 30 mares classified into healthy (n = 15) and endometritis (n = 15) based on their reproductive history, intrauterine fluid accumulation, gross appearance of LVL samples, endometrial cytology and bacterial culture. The samples were subjected to 16S rRNA sequencing. RESULTS: Notable differences in the uterine microbiome were observed between healthy mares and mares with endometritis at various taxonomic levels. In healthy mares, the most abundant phylum, class, order and family were Firmicutes, Bacilli, Bacillales and Paenibacillaceae, respectively. In contrast, the most abundant corresponding taxonomic levels in mares with endometritis were Proteobacteria, Gammaproteobacteria, Enterobacterales and Enterobacteriaceae, respectively. At the genus level, Brevibacillus and Paenibacillus were more abundant in healthy mares, whereas Escherichia, Salmonella and Klebsiella were more abundant in mares with endometritis. In healthy mares, Brevibacillus brevis was the most abundant species, followed by Brevibacillus choshinensis and Paenibacillus sp JDR-2. However, in mares with endometritis, Escherichia coli was the most abundant species, followed by Salmonella enterica and Klebsiella pneumoniae. CONCLUSIONS: These results confirmed the previously reported presence of a uterine microbiome in healthy mares and helped unravel some alterations that occur in mares with endometritis. The findings can potentially help formulate new approaches to prevent or treat equine endometritis.

Correlation of testicular ultrasonography, testicular biometry, serum testosterone levels and seminal attributes in pre and post-pubertal age for breeding soundness evaluation in Osmanabadi bucks.

The present research work entitled "Correlation of testicular ultrasonography, testicular biometry, serum testosterone levels and seminal attributes in pre- and post-pubertal age for breeding soundness evaluation in Osmanabadi bucks" was undertaken in 18 healthy Osmanabadi bucks from the Instructional Livestock Farm Complex, Bombay Veterinary College, Mumbai, Maharashtra. The body weight (kg), scrotal circumference (cm) and testicular biometry (cm) of post-weaning 18 Osmanabadi male kids was recorded every 15 days from weaning, i.e., 120 ± 10 days along with serum testosterone (ng/ml) by radioimmunoassay method at monthly intervals for the next 6 months. Semen was collected six times on the seventh month onward during post-pubertal age at 15-day interval from 18 bucks. The semen was evaluated for macroscopic and microscopic tests. The body weight increased from 14.45 ± 0.67 to 19.57 ± 0.70 kg from four to nine and a half months of age. The average daily body weight gain was 31.27 g. Maximum body weight gain was 01.19 ± 0.16 kg from 5 to 6 followed by 01.15 ± 0.16 kg from 4 to 5 months of age. The scrotal circumference increased from 17.22 ± 0.56 to 19.03 ± 0.55 cm from four to nine and a half months of age with maximum increased between 4 and 5 followed by 6 and 7 months of age. The testicular length, width and thickness of right and left testicles were recorded by ultrasonography method. There was increase in mean right and left testicular length, width and thickness from 5.25 ± 0.19 to 5.84 ± 0.18 and 5.49 ± 0.21 to 6.16 ± 0.20; 2.99 ± 0.12 to 3.32 ± 0.12 and 3.10 ± 0.13 to 3.44 ± 0.12 and 2.97 ± 0.12 to 3.16 ± 0.12 and 3.06 ± 0.12 to 3.31 ± 0.11 cm, respectively by ultrasonography, between four to nine and a half months of age. Testicular length, width and thickness gain was at maximum in 5 to 6 months of age. Left testicular length was more than the right testis. Before puberty, there was sudden gain in body weight, testicular length and width. However, scrotal circumference showed significant increase after puberty. Body weight had highest correlation with ultrasonographic left testicular thickness (r = 1) followed by scrotal circumference, ultrasonographic right and left testicular width, left testicular length, right testicular length and thickness and least by right testicular thickness (r = 0.95). The semen was thin to thick in consistency and average semen density was 3.10 ± 0.05. Average semen volume was 0.81 ± 0.02 ml, mass activity, initial motility, live and dead sperm count, abnormal sperm count and sperm concentration were 3.45 ± 0.13, 76.16 ± 1.16 and 75.16 ± 1.28% and 24.84 ± 1.28, 12.30 ± 0.50% and 2631.04 ± 45.74 million/ml, respectively in 18 bucks in six collection at 15 days. There was significant rise in semen volume, mass activity, initial motility and concentration at 8.5 months and live count, density at 9 months of age which indicates the age of sexual maturity is 8.5 to 9 months in Osmanabadi bucks. The body weight had highest positive correlation with mass activity (r = 98) followed by initial motility, live sperm count and total sperm concentration, semen volume (r = 76). The scrotal circumference had highest positive correlation with initial motility (r = 98) followed by live sperm count, total sperm count, mass activity, semen volume (r = 86). On the other hand, body weight and scrotal circumference were negatively correlated with abnormal and dead sperm count. The mean testosterone concentration increased from 0.02 ± 0.004 to 5.75 ± 0.80 ng/ml between four and half to nine and half months of age, respectively. There was significant rise (p < 0.01) up to 1.38 ± 0.28 ng/ ml at 6.5 months, i.e., age of puberty and up to 5.75 ± 0.80 ng/ml at 9.5 months, i.e., age of sexual maturity. Testosterone had highest positive correlation with testicular length followed by testicular width, length, body weight and scrotal circumference, mass activity, live sperm count, initial motility, while it had highest negative correlation with dead and abnormal sperm count. From the present research work, it was concluded that the scrotal circumference, testicular length, width and thickness increased with increasing body weight. Before puberty, there was sudden gain in body weight, testicular length and width. However, scrotal circumference increased significantly at post-pubertal age. So testicular length, body weight, testicular width in pre pubertal age and scrotal circumference post-pubertal age can be used as indicator for selection of Osmanabadi bucks for breeding purpose. On the other hand, the semen parameters should consider only after 8.5 to 9 months of age for selection of Osmanabadi bucks for breeding.

Laparoscopic ovariectomy in dogs in late gestation.

BACKGROUND: Surgical sterilization of stray dogs is the most widely used technique to control stray dog population. Although ovariectomy is an effective technique for elective sterilization of female dogs, most stray dog population control programs generally utilize ovariohysterectomy for spaying female dogs. In the context of stray dog sterilization, laparoscopic ovariectomy has been utilized and reported to be cost-effective compared to programs utilizing open surgical approaches. However, when pregnant stray dogs are encountered either conventional ovariohysterectomy is performed or surgery is deferred altogether. It is reported that ovariectomy at any stage during canine pregnancy results in fetal resorption or abortion, however, clinical outcomes following laparoscopic ovariectomy in dogs in late gestation have not been previously reported. The purpose of this study was to investigate the outcome of laparoscopic ovariectomy with intra-gestational sac injection (IGSI) of potassium chloride (KCl) in dogs in late gestation. RESULTS: Eight client owned dogs in the gestational age range of 40-50 days underwent laparoscopic ovariectomy with IGSI of KCl. Laparoscopic ovariectomy resulted in decreased serum progesterone from 11.6 ± 2.6 ng/ml on day 0 to 1.3 ± 0.4 ng/ml 24 h' post-surgery. IGSI of KCl did not result in immediate fetal death and fetal death temporally closely followed the drop in serum progesterone noted 24 h post-ovariectomy. Viscous brown vulvar discharge preceded fetal expulsion by 12 h and all the fetuses were dead when expelled. Uterine evacuation was documented within 45 ± 20 h (1-3.5 days) in all dogs without any clinically significant complications. CONCLUSION: Laparoscopic ovariectomy enables elective termination of pregnancy and simultaneous neutering of dogs in late gestation and has potential applications in high volume stray dog sterilization programs which utilize laparoscopy. Utility of IGSI of KCl in this regard is unclear.