Culturable Microbial Population From the Upper Respiratory Tract of 1,010 Clinically Healthy Horses in Southern Brazil.

Microbiological diagnosis of equine respiratory infections is essential for disease management. However, reliable diagnosis can be a challenge due to colonization of the upper respiratory tract (URT) by a diverse microbial population, and because there is a lack of studies with samples from healthy animals. Aiming to guide adequate URT culture, this work reports culturable microbial population from the URT of 1,010 apparently healthy horses from 341 farms in Southern Brazil and identifies the putative presence of pathogenic microorganisms. Nasal swabs were cultured in 5% blood agar, and the isolates were phenotypically identified to genus level. A diverse respiratory microbial population was found, mostly composed of Gram-positive bacteria, including Staphylococcus spp., Bacillus spp., Streptococcus spp. and Corynebacterium spp. The microbiological profile from the nasal cavity of 911 horses was described, with the five most common profiles being: (1) Staphylococcus sp. + Gram-negative bacilli (12.67%), (2) Staphylococcus sp. in pure culture (12.47%), (3) Staphylococcus sp. + Bacillus sp. (10.10%), (4) Gram-negative bacilli in pure culture (6.93%), and (5) Staphylococcus sp. + Bacillus sp. + Gram-negative bacilli (6.73%). Streptococcus equi equi and Rhodococcus equi were detected in 34 horses (3.37%), demonstrating the presence of pathogenic bacteria along with commensal microorganisms in apparently healthy animals. The disclosed data may guide and facilitate microbiological diagnosis of URT infection in horses. The significant presence of Gram-negative bacilli was evidenced, as well as the occurrence of relevant pathogens, such as S. equi equi and R. equi, thus helping to improve diagnosis and antimicrobial therapy.

Prevalence of Streptococcus equi subsp. equi in horses and associated risk factors in the State of Rio Grande do Sul, Brazil.

The aim of this study was to estimate the prevalence of equine strangles and to identify associated risk factors for this disease through a cross-sectional study of nasal swabs. Nasal swabs (n=1010) from healthy equines (absence of nasal discharge, lymphadenopathy and cough) from 341 farms were plated on 5% blood agar; of these horses, 24 were identified as positive for Streptococcus equi through isolation, PCR and DNA sequencing. The estimated prevalence for individual animals was 2.3%, and for herds, it was 5.86%. Statistical analysis identified the following as associated risk factors: the number of group events that were attended by the equines (PR: 1.06); the sharing of food containers (PR: 3.74); and at least one previous positive diagnosis of strangles on the farm (PR: 3.20). These results constitute an epidemiological contribution to the horse industry and may support measures for the future control of the disease.

Embryonic development of in vitro matured and in vitro fertilized dog oocytes.

In several studies, early cleavage stage canine embryos have been derived from in vitro fertilized oocytes cultured under various conditions. Despite these results, IVF protocols for canine oocytes have yielded low fertilization rates. In this study, Experiment I compared the effects of tissue culture medium (TCM)-199 supplemented with either (A) 1 microg/ml estradiol or (B) 20 microg/ml estradiol + 1 microg/ml human somatotropin (hST) on the in vitro nuclear maturation rate of canine oocytes. Meiotic progression to the metaphase I and II (MI/MII) stages at 72 hr of in vitro culture (IVC) was 10.2% (11/108) in medium A versus 14.1% (30/142) in medium B (P = 0.802). In Experiment II, cleavage rate was determined among oocytes recovered from ovaries of bitches at different reproductive stages. Oocytes (n = 888) were retrieved from bitches at the follicular, anestrous, and luteal stages and selected for high morphological quality. Oocytes were matured for 48 hr in TCM-199 supplemented with 1 microg/ml hST + 20 microg/ml estradiol. Oocytes were in vitro fertilized with fresh canine spermatozoa that had been isolated on a Percoll gradient, and were cultured in synthetic oviduct fluid (SOF) medium with bovine serum albumin (BSA; 4 mg/ml) up to 5 days in 5% CO(2) in air at 37 degrees C. A proportion of oocytes (30.6%) with identifiable nuclear material had cytoplasm penetrated or fertilized by sperm. The percentage of oocytes developing into early stage embryos was 10.1% (27/267). Although pronuclear development was observed to be higher for oocytes recovered at the follicular phase, the cleavage rate was similar among oocytes recovered from bitches at the follicular, anestrus, and luteal stages. There was no correlation between the proportion of capacitated or acrosome reacted spermatozoa and pronuclei formation and/or percent cleavage. It was concluded that TCM-199 supplemented with 1 microg/ml hST and estradiol (20 microg/ml) supports nuclear and cytoplasmic maturation of canine oocytes. In this study, meiotic competence was verified by the in vitro production (IVP) and development of embryos up to the 8 cell-stage. Furthermore, the results indicate that, under the described conditions and despite the influence of reproductive status of the bitch on the developmental competence of in vitro fertilized oocytes to the pronuclei stage, cleavage was independent of donor's reproductive estrous cycle stage.