Canine brucellosis in a Saskatchewan kennel

Canine brucellosis is rare in Canada. This report describes an outbreak of Brucella canis infection within a kennel, emphasizing diagnostic and pathologic findings. Gender differences are described. The progestational, nongravid uterus, female spleen, and prostate gland are consistent sites of bacterial isolation.     

Contribution of AIDA-I to the pathogenicity of a porcine diarrheagenic Escherichia coli and to intestinal colonization through biofilm formation in pigs

In order to evaluate the role of the AIDA-I of porcine diarrheagenic Escherichia coli strain PD20 serogroup O143 (AIDA-I⁺, STb⁺), a mutant strain PD20M (AIDA-I⁻, STb⁺) was generated from strain PD20 by an allelic exchange procedure. In addition, the full-length aidA gene was reintroduced into strain PD20M to generate the complemented strain PD20C (pTaidA, AIDA-I⁺, STb⁺). A non-pathogenic E. coli strain PD71 was used as negative control. Each strain was inoculated to newborn pigs via stomach tube. Severity of diarrhea was evaluated clinically and intestinal colonization was assessed by histology, immunohistochemistry (IHC), and transmission electron microscopy (TEM) including immunogold electron microscopy (IGEM). The adhesion pattern to HeLa cells, bacterial auto-aggregation and biofilm formation were evaluated in vitro. Pigs infected with strains PD20 or PD20C developed diarrhea 16 and 28 h after inoculation, respectively, in contrast to pigs infected with strains PD20M or PD71. Histology, IHC, TEM and IGEM examinations showed heavy bacterial colonization with biofilm formation in the large intestine, and marked in vivo expression of AIDA-I protein in pigs infected with strains PD20 or PD20C in contrast to pigs infected with strains PD20M or PD71. The in vitro assays showed marked diffuse adherence to HeLa cells, enhanced bacterial auto-aggregation and significant biofilm formation (p < 0.05) by the AIDA-I⁺ strains, when compared to AIDA-I⁻ strains. These results demonstrate that expression of AIDA-I is essential for intestinal colonization and in vitro bacterial autoaggregation and biofilm formation. Thus, AIDA-I may be considered a significant virulence determinant in development of diarrhea caused by porcine diarrheagenic AIDA-I⁺ E. coli PD20 in piglets.     

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.     

Mastitis caused by Bacillus anthracis in a beef cow

A mixed-breed beef cow was presented with swelling of the front and hind left quarters of the mammary gland and mild depression. Direct examination and culture of the serosanguinous-like milk samples collected from these quarters were consistent with Bacillus anthracis infection.     

Systemic infection with Mortierella wolfii following abortion in a cow

Severe meningoencephalitis and endometritis associated with necrotizing vasculitis, thrombosis, and infarction were found at necropsy of a 4-year-old Aberdeen Angus cow with a history of abortion and neurological signs. Focal pyogranulomatous pneumonia and nephritis were also present. Fungal hyphae typical of zygomycetes were abundant within lesions, and Mortierella wolfii was cultured from multiple tissues. This is believed to be the first report of systemic mortierellosis following abortion in North America, and the second reported instance of encephalitis caused by M. wolfii in a cow.     

In vivo and in vitro colonization patterns of AIDA-I-positive Escherichia coli isolates from piglets with diarrhea.

Transmission electron microscopy (TEM) was used to characterize the colonization patterns of 3 pathogenic Escherichia coli strains: PD58 and PD149 of the AIDA-I/STb/EAST1 pathotype, serogroup O: ND (not determined), and PD31 of the LT/STb/EAST1 pathotype, serogroup O149. These strains were isolated from diseased piglets and caused diarrhea in experimentally inoculated, newborn, colostrum-deprived pigs. In this study, intestinal tissues from newborn pigs experimentally infected with a high inoculum (20 ml containing 10(10) cfu) were harvested and examined for bacterial colonization using light microscopy. A nonaqueous perfluorocarbon fixation method was used to preserve the glycocalyx of the microvillus border in tissues collected for TEM. Transmission electron micrographs revealed that E. coli strain PD149 displayed long flexible fimbria-like structures that intimately attached the bacteria both to the microvillus border of the upper colon and to adjacent bacteria. In vitro, this strain demonstrated the localized adherence pattern to HEp-2 cells characteristic of enteroaggregative E. coli (EAggEC). Both PD58 and PD31 strains colonized the upper colon through the formation of a biofilm, also characteristic of EAggEC. Strains PD58 and PD31 adhered poorly to HEp-2 cells in vitro, although these demonstrated a colonization pattern suggestive of diffuse and aggregative adherence, respectively. These findings suggest that strains PD58 and PD149, expressing the AIDA-I, factor and strain PD31 represents hybrid pathotypes of diarrheagenic E. coli and that they probably cause diarrhea in piglets through differing mechanisms.     

Isolation and association of Escherichia coli AIDA-I/STb, rather than EAST1 pathotype, with diarrhea in piglets and antibiotic sensitivity of isolates.

To identify emerging Escherichia coli that have the potential to cause diarrhea in pigs, the prevalence of E. coli pathotypes was determined among 170 and 120 isolates from diarrheic and nondiarrheic piglets, respectively. The isolates were tested for F4, F5, F6, F18, and F41 fimbriae, for E. coli attaching and effacing (EAE), porcine attaching and effacing-associated (Paa), and adhesin involved in diffuse adherence (AIDA-I) factors, for LT, STa, STb, and enteroaggregative heat-stable (EAST1) enterotoxins, and for Shiga toxins (Stxl, Stx2, and Stx2e), using DNA hybridization and polymerase chain reaction. All isolates were O-serotyped and tested for antibiotic resistance against 10 drugs. Seventeen different pathotypes, accounting for 40.0% of the isolates, were recovered from diarrheic piglets. The main pathotypes included EAST1 (13.5%), F4/LT/STb/EAST1 (6.5%), AIDA-I/STb/EAST1 (4.1%), F5/STa (2.9%), EAE/EAST1 (2.9%), and AIDA-I/F18 (2.3%). Only 3 pathotypes, EAE (11.7%), EAST1 (10.8%), and EAE/EAST1 (3.3%), were recovered from nondiarrheic piglets. Paa factor was detected in 8.8% and 7.5% of isolates from diarrheic and nondiarrheic piglets, respectively, and always was associated with other virulence determinants. Overall, 22.9% of isolates from diarrheic piglets appeared to be enteropathogens: enterotoxigenic E. coli (11.7%), enteropathogenic E. coli (3.5%), and E. coli isolates (3.0%) for which none of the above adherence factors was detected. Pathotypes AIDA-I/STb/EAST1 and AIDA-I/STb were isolated only from diarrheic piglets and accounted for 4.7% of isolates. Strains of these pathotypes induced diarrhea when inoculated into newborn colostrum-deprived pigs, in contrast to an isolate positive only for EAST1, which did not induce diarrhea. Antibiotic sensitivity test showed that isolates of the AIDA-I/STb/EAST1 and AIDA-I/STb pathotypes were the only strains sensitive to enrofloxacin, gentamicin, neomycin, and trimethoprim-sulfamethoxazole. This study showed that at least 20.5% of isolates from diarrheic piglets appeared to be associated with AIDA-I/STb pathotype and that EAST1 pathotype is probably not an important marker for diarrhea in piglets.     

Retrospective study on Escherichia coli infection in broilers subjected to postmortem examination and antibiotic resistance of isolates in Trinidad

An 8-yr retrospective study was conducted to evaluate the rate of Escherichia coli infection and antibiotic resistance of isolates from diseased broilers submitted for diagnosis in Trinidad from 1990 to 1997. Of a total of 906 cases of diseased birds subjected to postmortem examination, 603 (66.6%) had E. coli infection. The number of cases increased over the years from 16 in 1990 to a peak of 294 in 1996. For every year, at least 50% of all broiler cases had E. coli infection. The rate of infection was significantly higher during the rainy season (74.1 +/- 6.9%) than during the dry season (57.8 +/- 7.0%). Approximately 50% of all E. coli isolates were resistant to 9 out of a total of 11 antimicrobial drugs selected for the study. The isolates showed an increasing trend of resistance to amoxicillin, apramycin, gentamicin, nitrofurantoin, norfloxacin, and sulfamethoxazole-trimethoprim. However, only the trends of resistance to apramycin and norfloxacin were statistically significant. Overall, of the antimicrobial drugs selected, norfloxacin relatively appeared as the best choice for treatment. From this study, we conclude that the high rate of E. coli infection in broilers submitted for diagnosis along with the high resistance of isolates to antimicrobial drugs constitute a threat to the poultry industry on the island.