Fecal Shedding of Clostridioides difficile in Calves in Sao Paulo State, Brazil.

OBJECTIVE: This study aimed to evaluate the fecal shedding of C. difficile in calves on farms in Sao Paulo State, Brazil. MATERIALS AND METHODS: Fecal samples (n=300) were collected from diarrheic (n=78) and nondiarrheic (n=222) calves less than 60 days of age from 20 farms. Fecal samples were inoculated into enrichment broth supplemented with taurocholate and cultured under anaerobic conditions. Colonies suspected to be C. difficile were harvested for DNA extraction and then multiplex PCR for the detection of genes encoding toxins A and B and binary toxins. All toxigenic isolates were ribotyped and tested for antimicrobial susceptibility, and five selected strains were subjected to whole-genome sequencing to determine their sequence type. RESULTS AND DISCUSSION: C. difficile was isolated from 29.3% (88/300) of the samples. All toxigenic isolates (17/88, 19.3%) were classified as ribotypes RT046 (13/17 -79.47%, A+B+ CDT-) and RT126 (4/17=20.53%, A+B+ CDT+). The sequenced strains from RT046 were classified as ST35 (Clade 1), while those from RT126 were classified as ST11 (Clade 5). No associations between the epidemiological factors in any of the groups and C. difficile isolation were observed. Most of the toxigenic isolates (16/17=94.41%) were classified as multidrug-resistant. Calves can be an important source of toxigenic C. difficile strains, including multidrug-resistant isolates from ribotypes commonly observed in humans.

Hereditary myotonia in cats associated with a new homozygous missense variant p.Ala331Pro in the muscle chloride channel ClC-1.

Three-related cats were evaluated for a history of short-strided gait and temporary recumbency after startle. Neurological examination, electromyography (EMG), muscle biopsies, and a chloride voltage-gated channel 1 (CLCN1) molecular study were performed. Clinically, all 3 cats presented myotonia with warm-up phenomenon and myotonic discharges during EMG examination. Muscle biopsies showed normal muscle architecture and variation in the diameter of myofiber size with the presence of numerous hypertrophic fibers. The molecular study revealed a missense variant (c.991G>C, p.Ala331Pro) in exon 9 of the CLCN1 gene, responsible for the first chloride channel extracellular loop. This mutation was screened in 104 control phenotypically normal unrelated cats, and all were wildtype. The alanine at this position is conserved in ClC-1 (chloride channel protein 1) in different species, and 2 mutations at this amino acid position are associated with human myotonia. This is the third CLCN1 mutation described in the literature associated with hereditary myotonia in cats and the first in domestic animals located in an extracellular muscle ClC-1 loop.

Pleural Empyema in Six Horses: A Retrospective Case Series.

The clinical findings of pleural empyema in six horses were retrospectively studied using epidemiological, clinical pathology, microbiological, ultrasound, and post-mortem data. The clinical findings included tachycardia (n = 3/6), tachypnea (n = 6/6), cyanotic mucosa (n = 2/6), hyperthermia (n = 4/6), inspiratory or mixed dyspnea (n = 6/6), presence of fluid and/or pleural rubbing (n = 2/6) and coarse crackling on auscultation (n = 4/6). Horses demonstrated leukocytosis (16.22 × 103/µL) with neutrophilia (12.32 × 103/µL) and hyperfibrinogenemia (633.33 mg/dL) and an increase in urea (69.80 mg/kg) and globulins (5.22 g/dL). The pleural fluid exhibited exudate (n = 5/6). The pathogens isolated from transtracheal wash (TTW) and/or pleural effusion included Aspergillus fumigatus, Enterobacter cloacae, alpha and beta hemolytic Streptococcus, Pseudomonas aeruginosa, Salmonella sp., Streptococcus equi subsp. zooepidemicus, and beta hemolytic Staphylococcus. The in vitro tests of microbial sensitivity of the isolates revealed that ceftiofur (5/6) and penicillin (3/6) were the most effective drugs. The fatality rate was 83% (5/6). The main post-mortem finding was the presence of fibrin in the pleural cavity with adhesion between the parietal and visceral pleura. These results show that pleural empyema is a complex disease pathophysiology that is refractory to conventional treatment.

Sequential Cerebrospinal Fluid Sampling in Horses: Comparison of Sampling Times and Two Different Collection Sites.

Analysis of the cerebrospinal fluid (CSF) is important as a complementary test in horses with neurologic diseases, and sequential analysis may provide information about the treatment response or evolution and quantitative measures of the CSF drug concentration during treatment. The aim of this study was to compare erythrocyte and nucleated cell counts and protein concentration in multiple CSF samples obtained sequentially from two different puncture sites in clinically healthy horses. Eight and 12 horses, with no evidence of neurologic disease, were subjected to CSF collection from the atlanto-occipital (AO) and C1-C2 spaces, respectively. Cytologic and chemical analyses were performed on the CSF obtained at five sampling times (T1, T2, T3, T4, and T5). Repeated measures models were used to compare the mean erythrocyte count, nucleated cell count, and total protein concentration between the AO and C1-C2 groups at each sampling time. C1-C2 CSF had a significantly higher total protein concentration at T1 and T4 than that of AO CSF. All total protein concentration values remained within the reference interval (<90 mg/dL) for all sampling times and groups. No statistical difference was present between results at T2, T3, T4, and T5 and at T1 in both groups for all analyses. In conclusion, five consecutive AO or C1-C2 CSF collections with at least a 7-d interval did not result in alterations in the CSF erythrocyte and nucleated cell counts and total protein concentrations and did not interfere with the CSF analysis results.