Canine Fetal Peritonitis

Background: The occurrence of neonatal diseases and neonatal deaths in canines are considered higher in the first days of life of individuals and bacterial diseases are considered the second most common cause. Inflammation of the mesothelial lining of the abdominal cavity is referred peritonitis, which can be classified as primary or secondary and there is no description on the occurrence of canine fetal peritonitis in the veterinary literature. The objective of this study is to describe a case of canine fetus peritonitis, presenting abnormal sonographic findings of the fetus and abnormalities identified after birth that were not yet described in veterinary literature.Case: A female Poodle, six years of age, considered healthy and without clinical complications during the gestational period, was referred for sonographic evaluation at the end of the pregnancy, in order to evaluate of the fetal viability and estimate the gestational period. By ultrasound examination, two canine fetuses were identified and based on its morphogenesis and biparietal diameter, the estimated gestational age was approximately 58 days. On the individual examination of each of the fetus by ultrasound, it was found that one of them had normal sonographic findings and on the second canine fetus it was found an increase of the intra-abdominal echogenicity of tissues, loss of the sonographic definition of the intestinal loops and presence of laminar anechoic areas between the interfaces of organs, consistent with abdominal effusion. Parturition occurred the following day of the ultrasound examination no complications and with the birth of two canine fetuses. About six hours after birth, one of the neonates showed dyspnea, hypothermia, areas of ecchymosis, suffusion in the ventral abdominal region, icteric and pale mucous membrane and the presence of abdominal fluid and came to death about 30 min after. Cytological analysis of the sample revealed exuberant amount of erythrocytes, fibrin and leukocytes. Culture revealed infection by Staphylococccus aureus and Enterococcus sp. Presence of bloody fluid in the abdominal cavity was observed at necropsy, with yellowish parietal surface of the thoracic and abdominal cavities and petechiae on the lungs, heart, stomach surfaces and intestinal mucous membrane. Microscopically, the lungs showed extensive areas of parenchymal consolidation, vascular congestion and intense inflammatory infiltrate in the alveolar wall; the liver was characterized by vascular congestion, areas of degeneration of hepatocytes and presence of calcified foci, necrosis and inflammatory infiltrates. Associating the results and findings of the exams was possible to conclude the diagnostic of fetal peritonitis.Discussion: The visualization of abnormal findings in affected fetus by a gestational ultrasound exam makes this report inedited and relevant for veterinary obstetrics. This demonstrates the importance of ultrasound exam in pregnant bitches for early detection of maternal fetal diseases. Presence of abdominal fluid and increased mesenteric ochogenicity associated with loss of the intestinal loop definition corroborates with the clinical suspicion of peritonitis in fetus. Findings on the anatomopathological examination, cytology and culture allowed conclusion of the cause of death. Bacterial forms in the cytological smear supported the diagnostic suspicion of peritonitis. Gestational ultrasound exam performed in this report was essential for proper conduct of the pregnant bitch and viability of the fetuses, suggesting that the detected fetal abnormalities showed predictive value in the viability of diseased individual. This case report contributes with the description of canine fetal peritonitis

Prevalence of Clostridioides difficile in Canine Feces and Its Association with Intestinal Dysbiosis

The role of Clostridioides (C.) difficile as an enteropathogen in dogs is controversial. In humans, intestinal bile acid-dysmetabolism is associated with C. difficile prevalence. The relationship between fecal qPCR-based dysbiosis index (DI) and especially the abundance of bile acid-converting Clostridium hiranonis with the presence of C. difficile in dogs was explored across the following 4 cohorts: 358 fecal samples submitted for routine diagnostic work-up, 33 dogs with chronic enteropathy, 14 dogs with acute diarrhea, and 116 healthy dogs. Dogs that tested positive for C. difficile had significantly higher DI (median, 4.4 (range from 0.4 to 8.6)) and lower C. hiranonis (median, 0.1 (range from 0.0 to 7.5) logDNA/g) than dogs that tested negative for C. difficile (median DI, −1 (range from −7.2 to 8.9); median C. hiranonis abundance, 6.2 (range from 0.1 to 7.5) logDNA/g; p < 0.0001, respectively). In 33 dogs with CE and 14 dogs with acute diarrhea, the treatment response did not differ between C. difficile-positive and -negative dogs. In the group of clinically healthy dogs, 9/116 tested positive for C. difficile, and 6/9 of these had also an abnormal DI. In conclusion, C. difficile is strongly linked to intestinal dysbiosis and lower C. hiranonis levels in dogs, but its presence does not necessitate targeted treatment

Correlation between Targeted qPCR Assays and Untargeted DNA Shotgun Metagenomic Sequencing for Assessing the Fecal Microbiota in Dogs

DNA shotgun sequencing is an untargeted approach for identifying changes in relative abundances, while qPCR allows reproducible quantification of specific bacteria. The canine dysbiosis index (DI) assesses the canine fecal microbiota by using a mathematical algorithm based on qPCR results. We evaluated the correlation between qPCR and shotgun sequencing using fecal samples from 296 dogs with different clinical phenotypes. While significant correlations were found between qPCR and sequencing, certain taxa were only detectable by qPCR and not by sequencing. Based on sequencing, less than 2% of bacterial species (17/1190) were consistently present in all healthy dogs (n = 76). Dogs with an abnormal DI had lower alpha-diversity compared to dogs with normal DI. Increases in the DI correctly predicted the gradual shifts in microbiota observed by sequencing: minor changes (R = 0.19, DI 2, DI > 5, and DI > 8, respectively), compared to dogs with a normal DI (DI < 0, all targets within the RI), as higher R-values indicated larger dissimilarities. In conclusion, the qPCR-based DI is an effective indicator of overall microbiota shifts observed by shotgun sequencing in dogs

Intestinal Permeability in Dogs with Chronic Enteropathy

Dogs with chronic clinical signs of gastrointestinal disease such as diarrhea, vomiting, weight loss, and/or loss of appetite are often diagnosed with chronic enteropathy after infectious, parasitic, neoplasia, and extra-intestinal diseases have been excluded. Chronic enteropathy is a multifactorial disease for which the exact pathophysiology is not well elucidated, but it is believed that dysfunction of the intestinal barrier is involved, leading to increased intestinal permeability. The study aims were to, measure serum iohexol concentrations after oral ingestion of iohexol, measure serum lipopolysaccharide concentrations, evaluate the expression of apical junction complex proteins: zonula occludens-1, e-cadherin, claudin-1, and occludin in small intestine tissue by immunohistochemistry, and perform untargeted small RNA-sequencing of fecal samples of healthy dogs and dogs with chronic enteropathy. Serum iohexol was increased in dogs with chronic enteropathy, more specifically in a subset of dogs with immunosuppressive-responsive enteropathy, classified based on treatment-response. Serum iohexol concentrations correlated with the canine chronic enteropathy activity index. E-cadherin immunostaining intensity in the small intestine was significantly increased in dogs with chronic enteropathy and correlated with serum iohexol concentrations. Occludin, claudin-1, and ZO-1 immunostaining intensity was not different between groups in this study population. Untargeted fecal small RNA-sequencing revealed 10 novel differentially expressed microRNAs in dogs with chronic enteropathy compared to healthy dogs. Among these, miR-21, miR-182, miR-184, miR- 200b, miR-221, and miR-320 were previously reported to correlate with gastrointestinal disorders or neoplasia in humans. miR-29a, which was upregulated in dogs with chronic enteropathy in this study, was previously reported to be upregulated in both humans and in pigs and described as an intestinal permeability regulator. Lastly, miR-Let7b, miR-8807, and miR-8842 are novel miRNAs that have not previously been studied in the human or veterinary fields. Interestingly, miR-8807 expression was increased in dogs with immunosuppressive-responsive enteropathy when compared to dogs with food-responsive enteropathy or healthy control dogs. These results demonstrate that a subset of dogs with chronic gastrointestinal disease have altered intestinal permeability. Additional studies are needed to elucidate whether this dysfunction of the intestinal barrier plays an active role or is another sequalae to chronic gastrointestinal disease in dogs

Erratum to: Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition) (Autophagy, 12, 1, 1-222, 10.1080/15548627.2015.1100356

non present