Rapid Salmonella detection in experimentally inoculated equine faecal and veterinary hospital environmental samples using commercially available lateral flow immunoassays.

REASONS FOR PERFORMING STUDY: Salmonella enterica is the most commonly reported cause of outbreaks of nosocomial infections in large animal veterinary teaching hospitals and the closure of equine hospitals. Rapid detection may facilitate effective control practices in equine populations. Shipping and laboratory testing typically require ≥48 h to obtain results. Lateral flow immunoassays developed for use in food-safety microbiology provide an alternative that has not been evaluated for use with faeces or environmental samples. OBJECTIVES: We aimed to identify enrichment methods that would allow commercially available rapid Salmonella detection systems (lateral flow immunoassays) to be used in clinical practice with equine faecal and environmental samples, providing test results in 18-24 h. STUDY DESIGN: In vitro experiment. METHODS: Equine faecal and environmental samples were inoculated with known quantities of S. enterica serotype Typhimurium and cultured using 2 different enrichment techniques for faeces and 4 enrichment techniques for environmental samples. Samples were tested blindly using 2 different lateral flow immunoassays and plated on agar media for confirmatory testing. RESULTS: In general, commercial lateral flow immunoassays resulted in fewer false-negative test results with enrichment of 1 g faecal samples in tetrathionate for 18 h, while all environmental sample enrichment techniques resulted in similar detection rates. The limit of detection from spiked samples, ∼4 colony-forming units/g, was similar for all methods evaluated. CONCLUSIONS: The lateral flow immunoassays evaluated could reliably detect S. enterica within 18 h, indicating that they may be useful for rapid point-of-care testing in equine practice applications. Additional evaluation is needed using samples from naturally infected cases and the environment to gain an accurate estimate of test sensitivity and specificity and to substantiate further the true value of these tests in clinical practice.

Detection of different serotypes of Salmonella enterica in experimentally inoculated equine fecal samples by commercially available rapid tests.

BACKGROUND: Salmonella enterica can significantly impact management of animal facilities. Comprehensive screening is essential for effective control in high-risk populations. Availability of reliable point-of-care diagnostic tests would facilitate these efforts. HYPOTHESIS/OBJECTIVES: Compare the ability of commercially available rapid diagnostic assays (2 lateral flow immunoassays [LFIs], DNA hybridization [DNAH], real-time PCR [qPCR]), and culture to detect common serotypes of S. enterica in feces. ANIMALS: n/a. METHODS: In an experimental study, 112 S. enterica isolates were randomly selected from the 10 most common serotypes recovered at a veterinary hospital. Archived isolates were amplified in broth and standardized inocula (100 colony forming units) were incubated with equine feces in tetrathionate broth (TET). Cultures were tested in a blinded fashion by using LFIs, DNAH, qPCR, and culture. RESULTS: The LFIs detected 84% and 67% of isolates, respectively, but reactivity varied among serotypes. Both reacted poorly with serotype Cerro (Group K) isolates, and 1 LFI did not react with any serotype Mbandaka (Group C1) or Montevideo (Group C1) isolates. DNAH detected 94% of isolates, whereas culture and qPCR most reliably detected all serotypes. False-positive results were obtained for 4 negative controls by using DNAH and 1 negative control by using qPCR, but LFIs and culture had no false-positive results. CONCLUSIONS AND CLINICAL IMPORTANCE: Culture, qPCR, and DNAH were effective in detecting most Salmonella isolates, but have limited application at point-of-care settings. LFIs are appealing as point-of-care tests because of low cost and ease of use, but limited detection of some serotypes needs to be evaluated with samples obtained from naturally infected animals.

Environmental Salmonella in agricultural fair poultry exhibits in Colorado.

Salmonella enterica is a common zoonotic pathogen in humans. Transmission typically occurs through consumption of contaminated food products or contact with infected animals, including poultry or their environment. The objective of this study was to estimate the frequency of Salmonella contamination in the environment in poultry exhibits at agricultural fairs. Samples were collected from cages, feed, floors and tables in the exhibit and cultured for Salmonella. At least one environmental sample was positive for Salmonella in 10 of 11 fairs (91%), and Salmonella was isolated from 28 of 55 environmental samples (50.9%). Eleven different serotypes were detected. Results of this study demonstrate that environmental surfaces at agricultural fairs can be contaminated with Salmonella and could potentially serve as a route of transmission to bird owners and the general public. Poultry owners and the general public should be educated about the risks of Salmonella infection from the poultry exhibit environment. Agricultural fairs should consider instituting policies and practices to improve hygiene and mitigate the risk of zoonotic salmonellosis.

Comparison of two sampling and culture systems for detection of Salmonella enterica in the environment of a large animal hospital.

REASONS FOR PERFORMING STUDY: Nosocomial salmonellosis is an important problem in veterinary hospitals that treat horses and other large animals. Detection and mitigation of outbreaks and prevention of healthcare-associated infections often require detection of Salmonella enterica in the hospital environment. OBJECTIVES: To compare 2 previously published methods for detecting environmental contamination with S. enterica in a large animal veterinary teaching hospital. STUDY DESIGN: Hospital-based comparison of environmental sampling techniques. METHODS: A total of 100 pairs of environmental samples were collected from stalls used to house large animal cases (horses, cows or New World camelids) that were confirmed to be shedding S. enterica by faecal culture. Stalls were cleaned and disinfected prior to sampling, and the same areas within each stall were sampled for the paired samples. One method of detection used sterile, premoistened sponges that were cultured using thioglycolate enrichment before plating on XLT-4 agar. The other method used electrostatic wipes that were cultured using buffered peptone water, tetrathionate and Rappaport-Vassiliadis R10 broths before plating on XLT-4 agar. RESULTS: Salmonella enterica was recovered from 14% of samples processed using the electrostatic wipe sampling and culture procedure, whereas S. enterica was recovered from only 4% of samples processed using the sponge sampling and culture procedure. There was test agreement for 85 pairs of culture-negative samples and 3 pairs of culture-positive samples. However, the remaining 12 pairs of samples with discordant results created significant disagreement between the 2 detection methods (P<0.01). CONCLUSIONS: Persistence of Salmonella in the environment of veterinary hospitals can occur even with rigorous cleaning and disinfection. Use of sensitive methods for detection of environmental contamination is critical when detecting and mitigating this problem in veterinary hospitals. These results suggest that the electrostatic wipe sampling and culture method was more sensitive than the sponge sampling and culture method.

Efficacy of intranasal administration of a modified live feline herpesvirus 1 and feline calicivirus vaccine against disease caused by Bordetella bronchiseptica after experimental challenge.

BACKGROUND: Studies suggest that intranasal vaccination can stimulate nonspecific immunity against agents not contained within the vaccine, but this effect is not reported for cats. HYPOTHESIS: A modified live feline herpesvirus-1 (FHV-1) and feline calicivirus (FCV) intranasal vaccine will reduce clinical signs of disease caused by experimental infection with Bordetella bronchiseptica. ANIMALS: Twenty specific pathogen-free 12-week-old kittens. METHODS: Experimental study. Cats were randomized into 2 groups of 10 cats each. The vaccinated group was administered a single intranasal dose of a commercially available vaccine containing modified live strains of FHV-1 and FCV, and the control group remained unvaccinated. All 20 cats were administered B. bronchiseptica by nasal inoculation 7 days later and were observed daily for clinical signs of illness for 20 days. RESULTS: In the first 10 days after B. bronchiseptica challenge, vaccinated cats were less likely to be clinically ill than control cats with a median clinical score of 0/180 (range 0-5) versus 2/180 (range 0-8) (P = .01). Nine of 10 control cats and 2 of 10 vaccinated cats were recorded as sneezing during days 1-10 after challenge (P = .006). CONCLUSIONS AND CLINICAL IMPORTANCE: Intranasal vaccination against FHV-1 and FCV decreased signs of illness due to an infectious agent not contained in the vaccine. This nonspecific immunity could be beneficial for protection against organisms for which vaccines are not available and as protection before development of vaccine-induced humoral immunity.