Pathotypic and molecular characterization of a fowl adenovirus associated with inclusion body hepatitis in Saskatchewan chickens

Inclusion body hepatitis (IBH) is one of the major global disease problems, causing significant economic losses to poultry industry of the United States and Canada. The disease is characterized by its sudden onset and high mortalities. Amongst different serotypes of fowl adenoviruses (FAdVs) associated with IBH, serotype 8 of group I FAdV has been isolated from majority of IBH cases. In present studies, we isolated a FAdV from morbid liver of a 17-day-old broiler from a Saskatchewan broiler farm. This newly isolated virus was designated as IBHV(SK). However, based on the sequence analysis of the L1 region of the hexon gene, the IBHV(SK) may be classified as FAdV 8b strain 764. These studies describe for the first time the complete hexon gene sequence of FAdV serotype 8b. Experimental infection of 2-day-old (n = 48) and 2-wk-old (n = 56) chicks caused 83% and 43% mortalities, respectively. Determination of the complete hexon gene sequence of IBHV(SK) with establishment of a disease model in chickens will facilitate the development of type-specific diagnostic reagents and assays for the evaluation of potential experimental vaccines against pathogenic FAdV infections.     

Antimicrobial susceptibility of hazard analysis critical control point Escherichia coli isolates from federally inspected beef processing plants in Alberta, Saskatchewan, and Ontario

A survey to estimate the prevalence of antimicrobial resistance in Escherichia coli was conducted in 7 Canadian federally inspected processing plants during 2001. Escherichia coli isolates were recovered during routine hazard analysis critical control point sampling from beef carcasses and trim and subsequently tested for their antimicrobial susceptibility by using susceptibility panels. Of the 2653 isolates analyzed, 68% were sensitive to all 18 antimicrobials tested. For 14 of the 18 antimicrobials evaluated, the percentage of resistant isolates was < or = 1. Twenty-five percent of the isolates were resistant to tetracycline, 9% to sulfamethoxazole, 7% to streptomycin, and 3% to ampicillin. Multiple resistance was found in 12% of the isolates, with 7% showing resistance to 2 antimicrobials, 2% to 3 antimicrobials, 2% to 4 antimicrobials, and 1% to 5 or more antimicrobials. Forty-five different antimicrobial resistance patterns were observed. The reasons for the development of the antimicrobial resistance were not investigated in this study. This study was useful as a pilot to help to develop a national antimicrobial resistance surveillance program in Canada. This study indicates that laboratory standardization is possible for consistent results across the country and that the indicator organism, E. coli, is fairly easy to obtain for surveillance but Salmonella are not, due to their low prevalence in beef.     

Histopathologic and bacteriologic evaluations of cellulitis detected in legs and caudal abdominal regions of turkeys

The objective of this study was to identify the causative agent of cellulitis in turkeys. Eighteen flocks from nine producers were sampled at the local processing plant, and 37 birds with cellulitis on legs or caudal thoracic area were obtained. None of the 37 birds with cellulitis had lesions in other organs. On gross examination, lesions were categorized into two groups: cellulitis with unopened skin lesions (type a) and cellulitis with opened skin lesions (type b). Histopathologically, cellulitis with unopened skin lesions had dermal necrosis with underlying fibrin and inflammatory exudate but cellulitis with open skin lesions had chronic granulomatous/granulation tissue-type reaction associated with foreign material. A complete bacteriologic study was conducted on 25 of 37 birds. Bacteria were isolated from 12 of the 25 birds with cellulitis lesions. No aerobic, microaerophilic, or anaerobic bacteria were isolated from the remaining 13 birds with cellulitis lesions. Escherichia coli was isolated in low numbers in mixed cultures with Proteus mirabilis, Lactobacillus spp., Klebsiella spp., and Staphylococcus spp. in 9 of 12 lesions. The remaining few cases yielded P. mirabilis in pure culture or in mixed culture with Pseudomonas aeruginosa. Types a and b cellulitis lesions in turkeys could be associated with primary contact dermatitis and skin abrasions, respectively. Their occurrence is likely associated with different management practices.     

Attenuated cytokine responses in porcine lymph node cells stimulated with CpG DNA are associated with low frequency of IFNalpha-producing cells and TLR9 mRNA expression

The immune stimulatory effects of synthetic CpG DNA, on porcine peripheral blood mononuclear cells (PBMC) have been reported, but little is known about CpG-induced responses in other lymphoid tissues of pigs. We investigated innate immune responses induced by CpG DNA in cells from blood, lymph nodes (LN) and spleens of pigs. Porcine PBMC and lymph node cells (LNC) were stimulated in vitro with three classes (A-, B- and C-class) of CpG oligodeoxynucleotides (ODNs), and a non-CpG control ODN. All three classes of CpG ODNs induced significant production of IFNalpha, TNFalpha, IL-1, IL-6 and IL-12 in PBMC. In contrast, in LNC, only IL-12 was stimulated by all three classes of CpG ODNs, while IFNalpha, and IL-6 were induced by A- and C-class ODNs. No TNFalpha was induced in LNC by any of the ODNs. Significant lymphocyte proliferation was induced in PBMC by all three classes of CpG ODNs and non-CpG control. However, in LNC, B- and C-class ODNs induced significant proliferation, while no proliferation was seen with A-class and non-CpG control ODN. All three classes of ODNs induced NK-like cytotoxicity in PBMC and spleen cells, but were less effective in inducing NK cytotoxicity in LNC. We then investigated the reasons for the relatively poor CpG-induced responses in LNC. Our investigations revealed that LNC had a lower frequency of IFNalpha-secreting cells and expressed low levels of TLR9 mRNA compared to PBMC. We conclude that the lower number of IFNalpha-secreting cells and receptor expression may contribute to the attenuated responses in LNC following stimulation with CpG ODN.     

Adaptation to a Fat-Supplemented Diet by Cutting Horses

In a crossover experiment, eight mature Quarter Horses were exercised by simulated cutting-horse training and fed either a control concentrate (C) or a 10% fat-supplemented concentrate (F) with Bermuda grass hay in a 60:40 ratio. The experiment consisted of two 28-day experimental periods with a 28-day standardization period between treatments. Digestion trials and standardized exercise tests (SET) were conducted every 7 days during experimental periods. Dietary fat had no detrimental effect on digestion of diet constituents, and all digestion coefficients were within expected ranges. Likewise, there was no significant difference in heart rates between treatments or SET days. Plasma lactic acid concentrations did not differ (P > .05) between treatments, however, there was a decline (P < .05) with advancing SET day. When horses were fed fat, there was an increase (P < .05) in muscle glycogen storage from days 0 to 28, but there was no increase (P > .05) in muscle glycogen over time when horses were fed C. On days 7, 14, and 28, muscle glycogen utilization increased (P < .05) from day 0 when horses were fed fat but not C. Pre-exercise data were normalized such that day 0 values equaled zero, due to higher (P < .05) glycogen concentrations for C than F on day 0. Subsequently, muscle glycogen concentrations on day 28 before exercise were higher (P < .05) when horses were fed F than C. The results of this study indicate that Quarter Horses in simulated cutting-horse training can adapt to digestion of a fat-supplemented diet by day 14. However, it may take up to 28 days of adaptation to a fat-supplemented diet to observe increases in glycogen storage and use.     

Evaluating the Technique of Using Nitrogen Retention as a Response Criterion for Amino Acid Studies in the Horse

Six Quarter Horse yearling fillies were used in a duplicated 3 × 3 Latin square experiment to evaluate the technique of using nitrogen retention as a response criterion for amino acid studies in the horse. Yearlings were paired by age and randomly assigned to one of three concentrates fed with medium-quality Coastal Bermudagrass hay. Diet A, a soybean meal–based concentrate, was amino acid sufficient. Diet B, a cottonseed hull–based concentrate containing 2% urea, was amino acid deficient. Horses fed diet C, identical to diet B, were orally dosed with synthetic essential amino acids and cysteine, immediately before eating, to match amino acid levels in diet A. Nitrogen retention was not different between diets A and B. There was a trend for horses fed diet C to have higher nitrogen retention, as a result of higher nitrogen intake. Nitrogen retained as a percent absorbed was lower (P < .05) for diet B than for diet A, whereas values for diet C were not different from diets A or B. Nitrogen retention was not an effective response criterion in this study. Differences in nitrogen retained as a percentage absorbed were observed between amino acid–sufficient and deficient diets. However, horses fed amino acid–deficient diets and dosed with synthetic amino acids likely require some modified dosage level to achieve the same or higher values as those for amino acid–sufficient diets.