Salmonella infections in ninety Alberta swine finishing farms: serological prevalence, correlation between culture and serology, and risk factors for infection.

The aim of this study was to determine serological prevalence for Salmonella in 90 Alberta finishing swine farms over a 5-month period; to evaluate the correlation between the detection of Salmonella by bacteriological culture and serology; and to identify risk factors for Salmonella seroprevalence. Participating farms were visited 3 times. A total of 30 blood and 15 fecal samples were collected from finishing pigs on each farm. VetScreen Salmonella covalent mix-ELISA (Svanovir) and conventional culture were performed. The apparent Salmonella seroprevalences at the sample and farm level were 13.2% (95% confidence interval [CI], 10.5-15.5%) and 83.3% (95% CI, 74-90.4%), respectively. Most of the farms had within-farm seroprevalence of

Seroprevalence of and agroecological risk factors for Mycobacterium avium subspecies paratuberculosis and neospora caninum infection among adult beef cattle in cow-calf herds in Alberta, Canada.

A province-wide cross-sectional seroprevalence and agroecological risk factor study of Mycobacterium avium subspecies paratuberculosis (MAP) and Neospora caninum (NC) infection among cattle in 100 cow-calf herds in Alberta was conducted. The seroprevalence of MAP in adult cattle was 1.5% across all herds. Using a widely accepted herd test cutpoint of 2 or more seropositive cows out of 30 animals tested, 7.9% of herds were estimated to be infected (95% confidence interval (CI): 2.3-23.4%). Seroprevalence of MAP differed by agroecological region; specifically, cattle and herds in areas with high soil pH (> 7.0), southern latitudes, and arid climates had a moderately reduced risk of infection (P < 0.10). Seroprevalence of NC infection was 9.7% among adult beef cattle province-wide--these levels also varied by agroecological region--with 91.0% of herds infected overall.

Seroprevalence of Mycobacterium avium subspecies paratuberculosis, Neospora caninum, Bovine leukemia virus, and Bovine viral diarrhea virus infection among dairy cattle and herds in Alberta and agroecological risk factors associated with seropositivity.

A province-wide, cross-sectional seroprevalence and agroecological risk factor study of Mycobacterium avium subspecies paratuberculosis (MAP), Neospora caninum (NC), Bovine leukemia virus (BLV), and Bovine viral diarrhea virus (BVDv) genotypes 1 and 2 (BVDv1 and BVDv2) infection in dairy cattle herds in Alberta was conducted. Among adults, the seroprevalence of MAP, NC, and BLV was 9.1%, 18.5%, and 26.9%, respectively. For MAP, based on a herd test cutpoint of 2 or more seropositive cows, 58.8% of herds were infected. Herd-level seroprevalence for NC and BLV was 98.7% and 86.7%, respectively, based on a herd-test cutpoint of 1 seropositive cow. Among unvaccinated dairy heifers, the seroprevalence for BVDv1 and BVDv2 infection was 28.4% and 8.9%, respectively, while herd-level infection was 53.4% and 19.7%. Seroprevalence for MAP varied moderately by agroecological region, whereas that for NC, BLV, and BVDv1 and BVDv2 did not. For MAP, aridity and soil pH (correlated features of the region) were also important.

Longitudinal study of Salmonella species in 90 Alberta swine finishing farms.

The aim of this study was to determine the farm prevalence of Salmonella in 90 Alberta finishing swine farms over a 5-month period, to evaluate Salmonella distribution in the farm environment and to describe Salmonella serovar diversity on these farms. Ten veterinary practitioners selected 90 Alberta swine farms based on an annual production of > or =2000 market pigs per farm and the willingness of the producers to participate in the study. Between May and September 2000, twenty samples were collected from finishing swine and the environment of each farm. The annual production of selected farms represented approximately 25% of the market swine production in Alberta. Participating farms were geographically representative of major swine production areas in Alberta. Sixty (66.7%) farms had at least one Salmonella-positive sample, with confidence interval (CI) of 57.1-77.2%. Salmonella were detected in 14.3% of fecal and 20.1% of environmental samples. The number of Salmonella-positive samples per farm ranged from 1 to 19. Among environmental samples, Salmonella were most frequently recovered from boots (38.6%) and the main drain (31.8%). Twenty-two serovars were detected on the 60 Salmonella-positive farms. Serovars Typhimurium (78 isolates), Derby (71 isolates) and Infantis (47 isolates) were the most common. A single serovar was detected on 58 farms, while 2, 3 and >3 serovars were detected on 15, 10 and 7 farms, respectively. The Salmonella farm status changed frequently over the 5-month period indicating the dynamic nature of Salmonella infections on these farms.

Antimicrobial resistance of Salmonella isolated from finishing swine and the environment of 60 Alberta swine farms.

The study objective was to describe and evaluate antimicrobial resistance profiles in Salmonella isolated from Alberta swine finishing farms. Salmonella isolates (n = 322) were obtained from 192 fecal and 84 environmental samples of the 60 Salmonella-positive swine finishing farms. Isolates were classified susceptible, intermediate or resistant based on NCCLS guidelines. More than half of the isolates (53.4%) were susceptible to all of the 18 antimicrobials in the testing panel. No resistance was observed to amikacin, amoxicillin/clavulanic acid, cefoxitin, ceftiofur, ceftriaxone, cephalothin, ciprofloxacin, imipenem or nalidixic acid. Less than 1% of isolates were resistant to apramycin, gentamicin and trimethoprim/sulfamethoxazole. Higher frequencies of resistance were observed for chloramphenicol (4.7%), ampicillin (7.8%), kanamycin (11.8%), sulfamethoxazole (21.1%), streptomycin (25.5%) and tetracycline (38.8%). Eleven Salmonella serovars had isolates with resistance to > or =3 antimicrobials. The most frequently resistant serovar was Salmonella Derby, with 27 (38.0%) isolates resistant to > or =3 antimicrobials, including resistance to five and six antimicrobials. An absence of resistance to cephalosporins and fluoroquniolones and a low proportion of isolates resistant to amikacin, amoxicillin/clavulanic acid, apramycin, gentamicin and trimethoprim/sulfamethoxazole are encouraging findings from public health and animal health perspectives. Frequent resistance observed for ampicillin, kanamycin, sulfamethoxazole, streptomycin and tetracycline, antimicrobials commonly used in veterinary medicine for decades, indicates an urgent need to utilize these antimicrobials more prudently if their benefits are to be preserved.

Evaluation of a covalent mix-enzyme linked immunosorbent assay for screening of Salmonella antibodies in pig serum.

In this study, a commercial Salmonella covalent mix-enzyme linked immunosorbent assay (ELISA) for serological detection of Salmonella infection in swine was evaluated by comparing it with the conventional fecal culture method and inter-laboratory proficiency testing, using a panel of sera tested in 5 laboratories from Europe and North America. Comparison with culture results showed that 88.5% of 26 culture-positive animals were ELISA positive, as were 55% of 60 animals from 2 culture-positive pig herds. Of 90 animals from 2 high health farms with no clinical symptoms of salmonellosis, 98.9% tested negative. The interlaboratory comparison study found a kappa value of 0.9 between our laboratory (using an automated system) and the manufacturer laboratory (using the manual method). Comparison of ELISA results from all 5 participating laboratories showed very good to excellent agreement, between 85% and 97.5%. We found this assay to be useful for the screening of antibodies against Salmonella present in swine serum. It agrees well with bacterial cultures, is reproducible, sensitive, specific, repeatable, and suitable for automation.

Evaluation of four DNA extraction methods for the detection of Mycobacterium avium subsp. paratuberculosis by polymerase chain reaction.

Polymerase chain reaction (PCR) has been widely used due to its high specificity, sensitivity, and rapid turn-around time. However, inhibitory factors may be co-extracted with the target nucleic acid that will hinder the performance of PCR. In this study, DNA extraction methods for Mycobacterium avium subsp. paratuberculosis were evaluated including rapid lysis, organic extraction, silica-based and magnetic particle-based (MagaZorb) technologies on bacterial cells, and spiked bovine feces. Efficiency of the extraction was determined by PCR end point titration with primers targeting the insertion sequence, IS900. Results of the end point titrations are identical for bacterial cells and spiked feces. Inhibition was observed in PCR with DNA isolated from spiked feces, and a 1/100 dilution was able to alleviate this problem with DNA extracted by MagaZorb. A 1/1000 dilution was required for the other three methods. MagaZorb proved to be more efficient at removing inhibitory factors and required the least labor and completion time. Further evaluation is required for its utilization in other clinical specimens.

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.

Mycobacterium paratuberculosis in dairy herds in Alberta.

Fifty dairy herds in Alberta were tested for the presence of Mycobacterium paratuberculosis by fecal culture and serum enzyme linked immunosorbent assay (ELISA). Individual sera (1500) were tested for antibodies to M. paratuberculosis by ELISA. Fecal samples were combined in pools of 3 (10 pools/herd) for a total of 500 pools that were cultured for M. paratuberculosis. Thirty cultures, including all 10 pools from 1 herd, were not readable due to fungal contamination. The remaining 470 cultures, representing 49 herds, yielded 16 positive pools (3.4% +/- 2.1%) from 10 herds (20.4% +/- 11.3%). The ELISA of each of the 1500 sera detected 105 (7.0% +/- 2.4%) positive sera and 20 (40.0% +/- 13.6%) positive herds, based on 2 or more individual positive sera in the herd. The true herd-level prevalence, as determined by ELISA, was 26.8% +/- 9.6%. The true herd-level prevalence, as determined by M. paratuberculosis fecal culture, ranged from 27.6% +/- 6.5% to 57.1% +/- 8.3%, depending on whether 1, 2, or all 3 individual fecal samples in the positive fecal pool were culture positive.

Prevalence of Salmonella in dairy herds in Alberta.

Fifty dairy herds in Alberta were tested for the presence of Salmonella. Four (8%) dairy herds had at least 1 cow shedding Salmonella. Different isolates were identified by serotyping, phage typing, and antibiotic resistance patterns. Pulsed-field gel electrophoresis patterns were determined for unique isolates.

Salmonella spp. shedding by alberta beef cattle and the detection of Salmonella spp. in ground beef.

Breeder cows, cattle recently arrived at feedlots, and cattle about to be shipped for slaughter were tested for Salmonella spp. No Salmonella spp. were detected in fecal samples from breeding cows. Nineteen of 1,000 (1.9%) fecal samples from recently arrived feedlot cattle were positive for Salmonella spp. compared to only 2 of 1,000 (0.2%) fecal samples taken within 2 weeks of slaughter. The positive fecal samples were collected in 5 of 50 (10%) "recent arrival" pens tested and in 1 of 50 (2%) pens tested within 2 weeks of slaughter. The serotypes isolated were Salmonella Agona, Salmonella Enteritidis, Salmonella Typhimurium DT104, and Salmonella 4,5,12:i:-. Ground beef samples purchased from retail outlets throughout Alberta were processed for Salmonella spp. Thirteen of 1,002 (1.3%) samples were positive for Salmonella spp. The serotypes isolated from ground beef were Salmonella Anatum, Salmonella Heidelberg, Salmonella Montevideo, Salmonella Typhimurium, Salmonella Typhimurium var. Copenhagen, and Salmonella Rough-O:i:1,2. The antibiotic resistance and pulsed-field electrophoresis gel macrorestriction patterns of all isolates were compared.