The objective of this study was to evaluate the effect of varying levels of hempseed meal supplementation on antibody and cell-mediated immune responses, as well as the expression of some of the important immunoregulatory cytokines. Treatments consisted of hempseed meal supplementation at 0 (control), 10, 20, and 30% of the total diet. Goats were randomly assigned to one of the four treatments n = 10. Cell-mediated immune response was evaluated on day 59 of the feeding period by measuring skinfold thickness at 24 h following intradermal injection of phytohemagglutinin. A significant increase in skinfold thickness was observed with increasing levels of supplementation as compared to that of the control group. Serum antibody titers to chicken ovalbumin were not significantly different between treatment groups. Cytokine concentrations of IL-6 increased linearly with increasing level of supplementation (p < 0.05), contrarily to the linear decrease that was observed for TNF-α (p < 0.05). Although IL-2 tended to increase with the 10 and 30% levels of supplementation (p < 0.07), the result was not significant, and no significant differences were obtained with respect to IL-4 concentrations. Cytokine gene expression values measured by RT-PCR, however, demonstrated some significant differences. HSM supplementation had no significant effect on the expression of IL-2 or IL-6. However, significant differences were observed with the 30% supplementation for IL-4 and TNF-α as compared to that of the control group (p < 0.05). IL-4 was down regulated for the 10 and 20% treatment groups but was upregulated for the 30% treatment group. TNF-α was downregulated in the 10% but upregulated for the 20 and 30% treatment groups. No significant differences were observed for the serum cortisol concentration or white blood cell counts. These results suggested that hempseed meal supplementation may improve cell-mediated immune response while having no effect on antibody-mediated immune response. However, more research needs to be conducted to determine the most efficacious inclusion rate.

The current study was designed to determine the prevalence of antimicrobial resistance and virulence genes in Escherichia coli isolated from canine diarrheal cases. Fecal samples were collected from 77 dogs with clinical diarrhea and 10 non-diarrheic dogs. Breed, age and sex, and clinical manifestations were recorded. Samples were plated on MacConkey and EMB agars, followed by standard isolation procedures and identification of E. coli. An antimicrobial susceptibility test using the disc diffusion method was performed against Ampicillin, Tetracycline, Trimethoprim, Gentamicin, and Streptomycin. PCR was used to determine if the isolates carry virulence intimin adherence protein (eaeA) and antimicrobial resistance genes. Body temperature, respiratory, and heart rates in dogs with diarrhea were significantly higher than in non-diarrheic dogs. PCR detected the eaeA virulence gene in 44(69.8%) of 63 isolates from diarrheic dogs. All isolates were resistant to ampicillin, and 55(87.3%), 47(74.6%), 29(46.0%), and 19(30.2%) of these were resistant to tetracycline, trimethoprim, gentamicin, and streptomycin, respectively. The frequency of antimicrobial resistance genes in the 63 isolates was 81.0, 52.4, 41.3, 33.3, 23.8 and 9.5% for CITM, tet (B), dfrA1, aac (3)-IV, aadA1 and tet (A), respectively. Overall, 6(9.4%), 16(25.4%), and 41(65.1%) were positive for one, two, three, or four resistance genes, respectively. In conclusion, the high prevalence of virulence (69.8%) and resistance 9.5-81.0% genes in E. coli isolates could be responsible for the diarrhea episodes, which may have posed therapeutic implications in affected dogs.