Investigation of procalcitonin and beta-defensin2 in the serum and feces of dogs with acute diarrhea.

BACKGROUND: Acute diarrhea is a common clinical condition where clinical parameters are used to assess disease severity, course, and prognosis. OBJECTIVES: The aim of this study was to investigate procalcitonin (PCT) and beta-defensin2 (Bdef2) as biomarkers for disease severity, course, and prognosis of dogs with acute diarrhea. METHODS: Dogs with acute diarrhea (enteritis group [EG], n = 35) were compared with 30 healthy controls. The dogs in the EG were scored using the Canine Acute Diarrhea Severity (CADS) index and grouped by bacterial fecal culture results. Procalcitonin and Bdef2 were analyzed in serum and feces. RESULTS: Dogs with acute diarrhea showed higher serum PCT concentrations (P < 0.0001) and lower fecal Bdef2 concentrations (P = 0.0001) than unaffected dogs. Serum PCT was moderately and positively related to the extent of disease classified by the CADS score. Dogs with Clostridium perfringens or hemolyzing Escherichia coli as predominant pathogen had increased serum Bdef2 concentrations (P < 0.01). Differentiation between uncomplicated (≤3 days) and complicated (>3 days) disease courses, determined by receiver operating characteristic (ROC) curves, resulted in a sensitivity of 0.74 and a specificity of 0.69 for serum PCT at a cutoff of 3.9 ng/mL. The serum PCT and fecal Bdef2 quotient resulted in a sensitivity of 0.80 and a specificity of 0.92, with a cutoff of 80.5. CONCLUSIONS: The results of the present study indicate that PCT and Bdef2 are potential biomarkers that can provide information on the severity, course, and prognosis of acute diarrhea in dogs.

Christmas disease in a Hovawart family resembling human hemophilia B Leyden is caused by a single nucleotide deletion in a highly conserved transcription factor binding site of the F9 gene promoter.

Hemophilia B is a classical monogenic, X-chromosomal, recessively transmitted bleeding disorder caused by genetic variants within the coagulation factor IX gene (F9). Although hemophilia B has been described in dogs, it has not yet been reported in the Hovawart breed. Here we describe the identification of a Hovawart family transmitting typical signs of an X-linked bleeding disorder. Five males were reported to suffer from recurrent hemorrhagic episodes. A blood sample from one of these males with only 2% of the normal concentration of plasma factor IX together with samples from seven relatives were provided. Next-generation sequencing of the mother and grandmother revealed a single nucleotide deletion in the F9 promoter. Genotyping of the deletion in 1,298 dog specimens including 720 Hovawarts revealed that the mutant allele was only present in the aforementioned Hovawart family. The deletion is located 73 bp upstream of the F9 start codon in the conserved overlapping DNA binding sites of hepatocyte nuclear factor 4α (HNF-4α) and androgen receptor (AR). The deletion only abolished binding of HNF-4α, while AR binding was unaffected as demonstrated by electrophoretic mobility shift assay using human HNF-4α and AR with double-stranded DNA probes encompassing the mutant promoter region. Luciferase reporter assays using wildtype and mutated promoter fragment constructs transfected into Hep G2 cells showed a significant reduction in expression from the mutant promoter. The data provide evidence that the deletion in the Hovawart family caused a rare type of hemophilia B resembling human hemophilia B Leyden.