Short communication: use of the BetaStar Plus assay for detection of ceftiofur antimicrobial residues in milk from individual cows following intramammary treatment for mastitis.

Development and use of on-farm assays to detect antimicrobial residues in milk is important to reduce the risk of violative residues in marketed milk. The objective of this study was to evaluate the effectiveness of a lateral-flow immunodiagnostic assay (BetaStar Plus, Neogen Corp., Lansing, MI) in detecting ceftiofur residues in milk from individual cows treated for mastitis. This assay is currently approved by the US Federal Drug Administration (FDA) for detecting ß-lactam residues in commingled milk. Forty-five dairy cows with clinical mastitis from 4 dairy farms were enrolled and treated intramammary with 125 mg of ceftiofur hydrochloride (Spectramast LC, Zoetis, Madison, NJ) according to the manufacturer's label recommendation. Composite milk samples were collected (A) before first intramammary antimicrobial treatment, (B) before the last intramammary antimicrobial treatment, (C) the last milking of the product-labeled milk withhold, (D) the first milking after the product-labeled milk withhold had been met, and (E) 72 h after the product-labeled milk withhold had been met. Samples were tested using the BetaStar Plus assay within 48 h of collection. Parallel samples were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and for somatic cell count and milk components. The BetaStar Plus assay identified 6.7, 60.0, 46.7, 22.2, and 6.7% positive samples at each of the respective time points. The assay had sensitivity and specificity of 100 and 84.7%, respectively, compared with liquid chromatography-tandem mass spectrometry analysis using FDA published residue tolerance levels for ceftiofur (or ceftiofur metabolites) as a threshold. The BetaStar Plus assay could be useful for detecting ceftiofur residues in milk from individual cows following intramammary treatment for mastitis before the milk is shipped for processing.

Prevalence of antimicrobial resistance among Salmonella on midwest and northeast USA dairy farms.

The objective of this study was to determine the prevalence of antimicrobial resistance among Salmonella isolated from dairy herds in New York, Minnesota, Michigan, and Wisconsin, USA. Serogroup and antimicrobial susceptibility characteristics were determined for Salmonella from cattle and environmental samples collected during August 2000-October 2001 as part of a longitudinal study where 129 herds were visited at 2-month intervals. Salmonella isolates were tested (using a broth microdilution method) for susceptibility to amoxicillin/clavulanic acid, ampicillin, ceftiofur, ceftriaxone, cephalothin, chloramphenicol, ciprofloxacin, gentamicin, kanamycin, nalidixic acid, streptomycin, sulfamethoxazole, tetracycline, and trimethoprim/sulfamethoxazole. Of the 1506 isolates tested for minimum inhibitory concentrations to these 14 antimicrobial agents, 81.2% were pan-susceptible and for most herds (81.6%) the predominant antimicrobial resistance pattern was pan-susceptible. At least 1 Salmonella isolate resistant to 5 or more antimicrobial agents was found on 23.6% of herds. This resistance phenotype was most common among serogroups B and E1 and among samples from calves and farmer-designated sick cows. Resistant samples most frequently exhibited resistance to tetracycline, streptomycin, and/or ampicillin. No samples were resistant to ceftriaxone (though 13 were in the intermediate range), and very few samples were resistant to ciprofloxacin (n=1), nalidixic acid (n=5), or trimethoprim/sulfamethoxazole (n=7).

Antimicrobial susceptibility of Salmonella from organic and conventional dairy farms.

The objective of this study was to compare antimicrobial susceptibility of Salmonella isolated from conventional and organic dairy farms in the Midwest and Northeast United States. Environmental and fecal samples were collected from organic (n = 26) and conventional (n = 69) farms in Michigan, Minnesota, New York, and Wisconsin every 2 mo from August 2000 to October 2001. Salmonella isolates (n = 1,243) were tested using a broth microdilution method for susceptibility to amoxicillin-clavulanic acid, ampicillin, ceftiofur, ceftriaxone, cephalothin, chloramphenicol, ciprofloxacin, gentamicin, kanamycin, nalidixic acid, streptomycin, sulfamethoxazole, tetracycline, and trimethoprim-sulfamethoxazole. Herd-level logistic regression and logistic proportional hazards multivariable models were used to examine the association between farm management type and susceptibility to antimicrobial agents. For most antimicrobial agents tested, susceptibility of Salmonella isolates was similar on organic and conventional herds when controlling for herd size and state. Conventional farms were more likely to have at least one Salmonella isolate resistant to streptomycin using logistic regression (odds ratio = 7.5; 95% confidence interval = 1.7-5.4). Conventional farms were more likely to have Salmonella isolates with greater resistance to streptomycin (odds ratio = 5.4; 95% confidence interval = 1.5-19.0) and sulfamethoxazole (odds ratio = 4.2; 95% confidence interval = 1.2-14.1) using logistic proportional hazards models. Although not statistically significant, conventional farms tended to be more likely to have at least one Salmonella isolate resistant to 5 or more antimicrobial agents when compared with organic farms.

Herd-level factors associated with isolation of Salmonella in a multi-state study of conventional and organic dairy farms II. Salmonella shedding in calves.

The objective of this study was to evaluate associations between herd-level factors and the isolation of Salmonella in calves from dairy farms in Minnesota, Wisconsin, Michigan and New York. Study farms were 129 conventional and organic farms enrolled without regard to previous history of Salmonella infection. Herds were sampled at 2-month intervals over a 1-year period. Salmonella was isolated in fecal samples from 176 (3.8%) of 4673 preweaned calves with 40 (31.0%) of 129 farms having at least one positive calf sample over the course of the study. Multivariable logistic regression using the generalized estimating equations approach was used to evaluate risk factors for Salmonella shedding after adjustment for effects of herd size, season, state of origin and the multiple sampling occasions per herd. Factors retained in the final model that were associated with an increased odds for Salmonella shedding were lack of routine feeding of milk replacer containing antimicrobials to preweaned calves (OR=2.8, 95% CI: 1.4, 5.8), use of maternity housing as a hospital area for sick cows more than once a month (OR=2.1, 95% CI: 1.1, 4.0), and cow prevalence level by visit, categorized into the following four-levels: > or =20% (OR=11.6, 95% CI: 5.7, 23.7), 10-19.9% (OR=4.7, 95% CI: 2.0, 11.5), 0.1-9.9% (OR=3.6, 95% CI: 1.5, 8.7) and 0% (reference level). Herd size was not associated with Salmonella shedding in the final multivariable model.

Herd-level factors associated with isolation of Salmonella in a multi-state study of conventional and organic dairy farms I. Salmonella shedding in cows.

The objective of this study was to evaluate associations between herd characteristics and the isolation of Salmonella from dairy cows in Minnesota, Wisconsin, Michigan, and New York. Study farms were 129 conventional and organic farms enrolled without regard to previous history of Salmonella infection. Herds were sampled at 2-month intervals over a 1-year period. This is the largest study to date on Salmonella shedding in dairy cows and the only study evaluating herd-level risk factors using longitudinal sampling to characterize Salmonella shedding on dairy farms. Salmonella was isolated in fecal samples from 1026 (4.9%) of 20,089 cows. Over the course of the study, 113 (87.6%) of 129 farms had at least one positive cow sample. Multi-variable logistic regression using the generalized estimating equations approach was used to test the association between herd-level risk factors and the dependent variable of within-herd prevalence by visit (number of Salmonella-positive cows/number of cows sampled) after adjustment for effects of herd size, season, state of origin, and the multiple sampling occasions per herd. Factors retained in the final model included lack of use of tiestall or stanchion facilities to house lactating cows (OR=1.9; 95% CI: 1.1-3.3), not storing all purchased concentrate or protein feeds in an enclosed building (OR=2.5; 95% CI: 1.3-4.9), not using monensin in weaned calf or bred heifer diets (OR=3.2; 95% CI: 2.0-5.4), access of lactating or dry cows to surface water (e.g., lake, pond, river, or stream) (OR=2.3; 95% CI: 1.3-3.9), disposal of manure in liquid form (slurry or irrigation, as opposed to disposal of manure by broadcast/solid spreader only) on owned or rented land (OR=1.8; 95% CI: 1.3-3.9), and cows eating or grazing of roughage from fields where manure was applied in solid or liquid form and not plowed under during the same growing season (OR=1.8; 95% CI: 1.0-3.0). A seasonal association was also present as cows were more likely to be Salmonella-positive in summer, spring, and fall compared to winter. Herd size was not associated with Salmonella shedding in the final multi-variable model. The herd-level risk factors identified in this study could potentially be implemented in Salmonella control programs on dairy farms.

Cattle and environmental sample-level factors associated with the presence of Salmonella in a multi-state study of conventional and organic dairy farms.

The objective of this study was to evaluate associations between cattle-level factors and environmental samples with the isolation of Salmonella from dairy farms in Minnesota, Wisconsin, Michigan, and New York. The study farms included 129 conventional and organic farms enrolled without regard to previous history of Salmonella infection. Herds were sampled at two-month intervals over a one-year period. Cattle groups more likely to be associated with Salmonella shedding (compared to preweaned calves) were cows designated as sick by farm personnel (OR=2.5, 95% CI: 1.7, 3.7), cows within 14 days of calving (OR=1.8, 95% CI: 1.1, 2.8), and cows due for culling within 14 days (OR=1.9, 95% CI: 1.0, 3.4). State of origin was also associated with the presence of Salmonella in samples from cattle and the farm environment; Midwestern states were more likely to have Salmonella-positive samples compared to New York. Cattle treated with antimicrobials within 14 days of sampling were more likely to be Salmonella-negative compared with nontreated cattle (OR=2.0, 95% CI: 1.1, 3.4). Farms with at least 100 cows were more likely to have Salmonella-positive cattle compared with smaller farms (OR=2.6, 95% CI: 1.4, 4.6). Season was associated with Salmonella shedding in cattle, and compared to the winter period, summer had the highest odds for shedding (OR=2.4, 95% CI: 1.5, 3.7), followed by fall (OR=1.9, 95% CI: 1.2, 3.1) and spring (OR=1.8, 95% CI: 1.2, 2.6). Environmental samples significantly more likely to be Salmonella-positive (compared to bulk tank milk) included, in descending order, samples from sick pens (OR=7.4, 95% CI: 3.4, 15.8), manure storage areas (OR=6.4, 95% CI: 3.5, 11.7), maternity pens (OR=4.2, 95% CI: 2.2, 8.1), haircoats of cows due to be culled (OR=3.9, 95% CI: 2.2, 7.7), milk filters (OR=3.3, 95% CI: 1.8, 6.0), cow waterers (OR=2.8, 95% CI: 1.4, 5.7), calf pens (OR=2.7, 95% CI: 1.3, 5.3), and bird droppings from cow housing (OR=2.4, 95% CI: 1.3, 4.4). Parity, stage of lactation, and calf age were not associated with Salmonella shedding.

Management practices and reported antimicrobial usage on conventional and organic dairy farms.

The primary objective was to compare reported antimicrobial usage between conventional and organic dairy farms. A secondary objective was to contrast selected management characteristics of conventional and organic dairy herds. A questionnaire was administered on site to selected dairy farmers located in Michigan, Minnesota, New York, and Wisconsin. Organic herds (n = 32) were smaller and produced less milk than conventional herds (n = 99). Lactating cows in organic dairies were more likely to be housed in tie stalls, whereas most conventional dairies housed cows in free stalls and milked in a parlor. Total mixed rations and purchased feeds were used on more conventional dairy farms compared with organic dairy farms. Conventional dairy producers were more likely to use advice from veterinarians for recommendations of treatment, and organic dairy producers were more likely to rely on advice from other farmers. Based on recall of antibiotic usage in the previous 60 d, 5.1, 84.9, 9.1, and 0.9% of farmers with conventional herds reported treatment of none, 1 to 10%, 11 to 25%, and >25% of milk cows, respectively. Most organic farmers (90.6%) reported no antibiotic treatments of milk cows, whereas 9.4% reported treating 1 to 10% of milk cows. Ceftiofur was the most commonly reported antibiotic for both farm types. Milk replacer containing antibiotics was reportedly used on 49.5% of conventional herds but only on one organic herd (3.1%). Antibiotics were used in heifer calves on 74.7% of conventional herds versus 21.9% of organic herds. Antibiotics to treat mastitis were used on 79.8% of conventional herds but on none of the organic herds. Most organic farms were in compliance with standards in advance of implementation of regulations.

Intramuscular administration of ceftiofur sodium versus intramammary infusion of penicillin/novobiocin for treatment of Streptococcus agalactiae mastitis in dairy cows.

OBJECTIVE: To determine the efficacy of intramuscular administration of ceftiofur sodium as treatment for intramammary infections attributable to Streptococcus agalactiae, compared with that for a standard treatment of intramammary infusion of penicillin/novobiocin. DESIGN: Prospective, randomized, controlled trial. ANIMALS: 72 lactating Holstein cows with intramammary infections caused by S agalactiae from 5 commercial dairies in Michigan. PROCEDURE: In 36 of 72 infected cows, ceftiofur was administered (2.2 mg/kg of body weight, IM, q 24 h) for 5 days; 150 mg of novobiocin and 100,000 U of procaine penicillin G was infused daily into each mammary gland of the other 36 cows for 2 days. Milk samples were collected aseptically at approximately 4 and 8 weeks after initial treatment. If cows were determined to be infected at 4 weeks after initial treatment, the treatment was repeated. RESULTS: The cure rate at 4 weeks (91.7%) and at 8 weeks (96.8%) after initial treatment for the penicillin/novobiocin-treated cows was significantly (P < 0.0001) higher, compared with that of the ceftiofur-treated cows (2.8 and 9.1%, respectively). Somatic cell counts were significantly (P < 0.0001) lower in the penicillin/novobiocin-treated group after treatment. CLINICAL IMPLICATIONS: Intramuscular administration of ceftiofur is not efficacious as a treatment to eliminate intramammary infections caused by S agalactiae and should not be used to reduce the prevalence of this organism in dairy herds.