Evidence of sheep and abattoir environment as important reservoirs of multidrug resistant Salmonella and extended-spectrum beta-lactamase Escherichia coli.

The increase in antimicrobial-resistant (AMR) foodborne pathogens, including E. coli and Salmonella in animals, humans, and the environment, is a growing public health concern. Among animals, cattle, pigs, and chicken are reservoirs of these pathogens worldwide. There is a knowledge gap on the prevalence and AMR of foodborne pathogens in small ruminants (i.e., sheep and goats). This study investigates the prevalence and antimicrobial resistance of extended-spectrum beta-lactamase (ESBL) E. coli and Salmonella from sheep and their abattoir environment in North Carolina. We conducted a year-round serial cross-sectional study and collected a total of 1128 samples from sheep (n = 780) and their abattoir environment (n = 348). Sheep samples consisted of feces, cecal contents, carcass swabs, and abattoir resting area feces. Environmental samples consisted of soil samples, lairage swab, animal feed, and drinking water for animals. We used CHROMAgar EEC with 4 µg/ml of Cefotaxime for isolating ESBL E. coli, and ESBL production was confirmed by double-disk diffusion test. Salmonella was isolated and confirmed using standard methods. All of the confirmed isolates were tested against a panel of 14 antimicrobials to elucidate susceptibility profiles. The prevalence of ESBL E. coli and Salmonella was significantly higher in environmental samples (47.7% and 65.5%) compared to the sheep samples (19.5% and 17.9%), respectively (P < 0.0001). We recovered 318 ESBL E. coli and 368 Salmonella isolates from sheep and environmental samples. More than 97% (310/318) of ESBL E. coli were multidrug-resistant (MDR; resistant to ≥3 classes of antimicrobials). Most Salmonella isolates (77.2%, 284/368) were pansusceptible, and 10.1% (37/368) were MDR. We identified a total of 24 different Salmonella serotypes by whole genome sequencing (WGS). The most common serotypes were Agona (19.8%), Typhimurium (16.2%), Cannstatt (13.2%), Reading (13.2%), and Anatum (9.6%). Prevalence and percent resistance of ESBL E. coli and Salmonella isolates varied significantly by season and sample type (P < 0.0001). The co-existence of ESBL E. coli in the same sample was associated with increased percent resistance of Salmonella to Ampicillin, Chloramphenicol, Sulfisoxazole, Streptomycin, and Tetracycline. We presumed that the abattoir environment might have played a great role in the persistence and dissemination of resistant bacteria to sheep as they arrive at the abattoir. In conclusion, our study reaffirms that sheep and their abattoir environment act as important reservoirs of AMR ESBL E. coli and MDR Salmonella in the U.S. Further studies are required to determine associated public health risks.

Changes in the prevalence of Salmonella serovars associated swine production and correlations of avian, bovine and swine-associated serovars with human-associated serovars in the United States (1997-2015).

As Salmonella enterica is an important pathogen of food animals, surveillance programmes for S. enterica serovars have existed for many years in the United States. Surveillance programmes serve many purposes, one of which is to evaluate alterations in the prevalence of serovars that may signal changes in the ecology of the target organism. The primary aim of this study was to evaluate changes in the proportion of S. enterica serovars isolated from swine over a near 20-year observation period (1997-2015) using four longitudinal data sets from different food animal species. The secondary aim was to evaluate correlations between changes in S. enterica serovars frequently recovered from food animals and changes in S. enterica serovars associated with disease in humans. We found decreasing proportions of S. enterica serovar Typhimurium, serovar Derby and serovar Heidelberg and increasing proportions of S. enterica serovar 4,[5],12:i:-, serovar Infantis and serovar Johannesburg in swine over time. We also found positive correlations for the yearly changes in S. enterica serovar 4,[5],12:i:-, serovar Anatum and serovar Johannesburg between swine and human data; in S. enterica Worthington between avian and human data; and in S. enterica serovar 4,[5],12:i:- between bovine and human data. We found negative correlations for the yearly changes in S. enterica serovar 4,[5],12:i:- and serovar Johannesburg between avian and human data.

Antimicrobial susceptibility of Clostridium difficile isolated from food animals on farms.

Clostridium difficile is commonly associated with a spectrum of disease in humans referred to as C. difficile-associated disease (CDAD) and use of antimicrobials is considered a risk factor for development of disease in humans. C. difficile can also inhabit healthy food animals and transmission to humans is possible. As a result of the complexity and cost of testing, C. difficile is rarely tested for antimicrobial susceptibility. A total of 376 C. difficile strains (94 each from swine and dairy feces, and 188 from beef cattle feces) were isolated from healthy food animals on farms during studies conducted by the National Animal Health Monitoring System. Using the Etest (AB Biodisk, Solna, Sweden), samples were tested for susceptibility to nine antimicrobials implicated as risk factors for CDAD (linezolid, amoxicillin-clavulanic acid, ampicillin, clindamycin, erythromycin, levofloxacin, metronidazole, rifampicin, and vancomycin). Vancomycin was active against all isolates of C. difficile (MIC90=3.0µg/ml) while almost all isolates (n=369; 98.1%) were resistant to levofloxacin. With the exception of vancomycin, resistance varied by animal species as follows: linezolid (8.5% resistance among swine versus 2.1 and 1.1% resistance among dairy and beef, respectively), clindamycin (56.4% resistance among swine versus 80% and 90.9% resistance among dairy and beef, respectively), and rifampicin (2.1% and 0% resistance among swine and dairy cattle isolates, respectively versus 14.3% resistance among beef isolates). Regardless of species, multiple drug resistance was observed most often to combinations of clindamycin and levofloxacin (n=195; 51.9%) and ampicillin, clindamycin and levofloxacin (n=41; 10.9%). The reason for the variability of resistance between animal species is unknown and requires further research.

Influence of commercial laying hen housing systems on the incidence and identification of Salmonella and Campylobacter.

The housing of laying hens is important for social, industrial, and regulatory aspects. Many studies have compared hen housing systems on the research farm, but few have fully examined commercial housing systems and management strategies. The current study compared hens housed in commercial cage-free aviary, conventional cage, and enriched colony cage systems. Environmental and eggshell pool samples were collected from selected cages/segments of the housing systems throughout the production cycle and monitored for Salmonella and Campylobacter prevalence. At 77 wk of age, 120 hens per housing system were examined for Salmonella and Campylobacter colonization in the: adrenal glands, spleen, ceca, follicles, and upper reproductive tract. All isolates detected from environmental swabs, eggshell pools, and tissues were identified for serotype. Two predominant Salmonella were detected in all samples:S.Braenderup andS.Kentucky.Campylobacter coli and C. jejuni were the only Campylobacter detected in the flocks. Across all housing systems, approximately 7% of hens were colonized with Salmonella, whereas >90% were colonized with Campylobacter Salmonella Braenderup was the isolate most frequently detected in environmental swabs (P<0.0001) and housing system impacted Salmonella spp. shedding (P<0.0001).Campylobacter jejuni was the isolate most frequently found in environmental swabs (P<0.01), while housing system impacted the prevalence of C. coli and jejuniin ceca (P<0.0001). The results of this study provide a greater understanding of the impact of hen housing systems on hen health and product safety. Additionally, producers and academia can utilize the findings to make informed decisions on hen housing and management strategies to enhance hen health and food safety.

Attribution of Salmonella enterica serotype Hadar infections using antimicrobial resistance data from two points in the food supply system.

A challenge to the development of foodborne illness prevention measures is determining the sources of enteric illness. Microbial subtyping source-attribution models attribute illnesses to various sources, requiring data characterizing bacterial isolate subtypes collected from human and food sources. We evaluated the use of antimicrobial resistance data on isolates of Salmonella enterica serotype Hadar, collected from ill humans, food animals, and from retail meats, in two microbial subtyping attribution models. We also compared model results when either antimicrobial resistance or pulsed-field gel electrophoresis (PFGE) patterns were used to subtype isolates. Depending on the subtyping model used, 68-96% of the human infections were attributed to meat and poultry food products. All models yielded similar outcomes, with 86% [95% confidence interval (CI) 80-91] to 91% (95% CI 88-96) of the attributable infections attributed to turkey, and 6% (95% CI 2-10) to 14% (95% CI 8-20) to chicken. Few illnesses (<3%) were attributed to cattle or swine. Results were similar whether the isolates were obtained from food animals during processing or from retail meat products. Our results support the view that microbial subtyping models are a flexible and robust approach for attributing Salmonella Hadar.

Microbiological impact of three commercial laying hen housing systems.

Hen housing for commercial egg production continues to be a societal and regulatory concern. Controlled studies have examined various aspects of egg safety, but a comprehensive assessment of commercial hen housing systems in the US has not been conducted. The current study is part of a holistic, multidisciplinary comparison of the diverse aspects of commercial conventional cage, enriched colony cage, and cage-free aviary housing systems and focuses on environmental and egg microbiology. Environmental swabs and eggshell pools were collected from all housing systems during 4 production periods. Total aerobes and coliforms were enumerated, and the prevalence of Salmonella and Campylobacter spp. was determined. Environmental aerobic and coliform counts were highest for aviary drag swabs (7.5 and 4.0 log cfu/mL, respectively) and enriched colony cage scratch pad swabs (6.8 and 3.8 log cfu/mL, respectively). Aviary floor and system wire shell pools had the greatest levels of aerobic contamination for all eggshell pools (4.9 and 4.1 log cfu/mL, respectively). Hens from all housing systems were shedding Salmonella spp. (89-100% of manure belt scraper blade swabs). The dry belt litter removal processes for all housing systems appear to affect Campylobacter spp. detection (0-41% of manure belt scraper blade swabs) considering detection of Campylobacter spp. was much higher for other environmental samples. Aviary forage area drag swabs were 100% contaminated with Campylobacter spp., whereas enriched colony cage scratch pads had a 93% positive rate. There were no differences in pathogen detection in the shell pools from the 3 housing systems. Results indicate egg safety is enhanced when hens in alternative housing systems use nest boxes. Additionally, current outcomes indicate the use of scratch pads in hen housing systems needs to be more thoroughly investigated for effects on hen health and egg safety.

Carriage of methicillin-resistant staphylococci by healthy companion animals in the US.

UNLABELLED: Antimicrobial-resistant staphylococci have been associated with wounded or ill companion animals, but little is known about the prevalence of resistant staphylococci among healthy animals. In this study, 276 healthy dogs and cats from veterinary clinics were tested for the presence of antimicrobial-resistant Staphylococcus spp. Isolates were tested for antimicrobial susceptibility and the presence of select resistance genes, and typed using Pulsed-Field Gel Electrophoresis (PFGE). Staphylococcus aureus and Staphylococcus pseudintermedius were also characterized using multilocus sequence typing (MLST), spa typing and SCCmec typing. Approximately 5% (14/276) of the animals were positive by enrichment for five species of staphylococci [Staph. aureus (n = 11), Staph. pseudintermedius (n = 4), Staphylococcus sciuri (n = 6), Staphylococcus simulans (n = 1) and Staphylococcus warneri (n = 1)]. Seventy-eight per cent (18/23) of staphylococci were resistant to oxacillin and also multidrug resistant (resistance to ≥ 2 antimicrobials). All Staph. aureus isolates were mecA+ and blaZ+, SCCmec type II, spa type t002, ST5 and clonal using PFGE. Staphylococcus pseudintermedius were SCCmec type IV or V, spa type t06 and ST170; two of the isolates were pvl(+) . These results suggest that healthy companion animals may be a reservoir of multidrug-resistant staphylococci, which may be transferred to owners and others who handle companion animals. SIGNIFICANCE AND IMPACT OF THE STUDY: In this study, antimicrobial-resistant coagulase-negative and coagulase-positive staphylococci were isolated from various body sites on healthy dogs and cats. Resistance to 14 antimicrobials was observed including resistance to oxacillin; the majority of staphylococci were also multidrug resistant. Results from this study suggest that healthy dogs and cats may act as reservoirs of antimicrobial-resistant bacteria that may be transferred to people by simple interaction with the animals. Such carriage poses an underlying risk of infection, which should be considered during handling of healthy dogs and cats by pet owners and veterinary personnel.

Sampling naturally contaminated broiler carcasses for Salmonella by three different methods.

Postchill neck skin maceration (NSM) and whole-carcass rinsing (WCR) are frequently used methods to detect salmonellae from processed broilers. These are practical, nondestructive methods, but they are insensitive and may result in false negatives (20 to 40%). Neck skin samples comprise only 4% of the skin from the broiler carcass by weight, while WCR will not detect firmly attached Salmonella organisms and only 7.5% of the rinsate is utilized. Whole-carcass enrichment (WCE) involves incubation of the whole carcass overnight in a preenrichment broth and can recover as few as 8 inoculated Salmonella cells per carcass. The objective of this study was to use NSM, WCR, and WCE sampling to detect naturally occurring Salmonella from the same commercially processed broiler either prechill or postchill. Ten carcasses were obtained prechill and another 10 postchill on each of two replicate days from each of two commercial processing plants. From each carcass, 8.3 g of neck skin was sampled, and then the carcass was rinsed with 400 ml of 1% buffered peptone water. Thirty milliliters was removed and incubated (WCR), and the remaining 370 ml of broth and the carcass were incubated at 37°C for 24 h (WCE). Overall, Salmonella organisms were detected on 21, 24, and 32 of 40 prechill carcasses by NSM, WCR, and WCE, respectively, while 2, 2, and 19 of 40 postchill carcasses were positive by the respective methods. Prechill carcasses were 64% (77 of 120) positive for Salmonella, while postchill carcasses were 19% (23 of 120) positive. Commercial processing reduced the positive-sample prevalence by 45%. Salmonella organisms were detected on 20% (24 of 120) of the samples from plant 1 and 63% (76 of 120) of the carcasses from plant 2. This study demonstrates significant differences in the results for Salmonella prevalence among sampling methods both before and after immersion chilling, as well as between processing plants on days that samples were taken.

Evidence for horizontal and vertical transmission in Campylobacter passage from hen to her progeny.

Campylobacter is an important human pathogen, and consumption of undercooked poultry has been linked to significant human illnesses. To reduce human illness, intervention strategies targeting Campylobacter reduction in poultry are in development. For more than a decade, there has been an ongoing national and international controversy about whether Campylobacter can pass from one generation of poultry to the next via the fertile egg. We recognize that there are numerous sources of Campylobacter entry into flocks of commercial poultry (including egg transmission), yet the environment is often cited as the only source. There has been an abundance of published research globally that refutes this contention, and this article lists and discusses many of them, along with other studies that support environment as the sole or primary source. One must remember that egg passage can mean more than vertical, transovarian transmission. Fecal bacteria, including Campylobacter, can contaminate the shell, shell membranes, and albumen of freshly laid fertile eggs. This contamination is drawn through the shell by temperature differential, aided by the presence of moisture (the "sweating" of the egg); then, when the chick emerges from the egg, it can ingest bacteria such as Campylobacter, become colonized, and spread this contamination to flock mates in the grow house. Improvements in cultural laboratory methods continue to advance our knowledge of the ecology of Campylobacter, and in the not-so-distant future, egg passage will not be a subject continuously debated but will be embraced, thus allowing the development and implementation of more effective intervention strategies.

Comparison of individual, pooled, and composite fecal sampling methods for detection of Salmonella on U.S. dairy operations.

The objectives of this study were to estimate the prevalence of Salmonella for individual, pooled, and composite fecal samples and to compare culture results from each sample type for determining herd Salmonella infection status and identifying Salmonella serovar(s). During the U.S. Department of Agriculture National Animal Health Monitoring System Dairy 2007 study, data and samples were collected from dairy operations in 17 major dairy states. As part of the study, composite fecal samples (six per operation) were collected from cow areas, such as holding pens, alleyways, and lagoons, where manure accumulates. Fecal samples also were collected from individual cows (35 per operation), and fecal sample pools were created by combining samples from 5 cows (7 per operation). A total of 1,541 composite fecal samples were collected from 260 operations in 17 states, and 406 (26.3%) of these samples were culture positive for Salmonella. Among the 116 operations for which all three sample types were obtained, 41.4% (48 operations) were Salmonella culture positive based on individual samples, 39.7% (46 operations) were positive based on pooled samples, and 49.1% (57 operations) were positive based on composite fecal samples. Relative to individual samples, the sensitivity of composite fecal samples for determining herd infection status was 85.4% and the sensitivity of pooled fecal samples was 91.7%. On 33.6% of operations (39 of 116), Salmonella was cultured from all three fecal sample types (individual, pooled, and composite), and 20 (51.3%) of these operations had exactly the same serovar in all three sample types. Use of composite fecal samples is less costly and time-consuming than use of individual or pooled samples and provides similar results for detecting the presence and identifying serovars of Salmonella in dairy herds. Therefore, composite sampling may be an appropriate alternative to culture of individual samples when assessing Salmonella status in dairy herds.

Characterization of extended-spectrum cephalosporin-resistant Salmonella enterica serovar Heidelberg isolated from food animals, retail meat, and humans in the United States 2009.

Salmonella enterica is one of the most common causes of foodborne illness in the United States. Although salmonellosis is usually self-limiting, severe infections typically require antimicrobial treatment, and ceftriaxone, an extended-spectrum cephalosporin (ESC), is commonly used in both adults and children. Surveillance conducted by the National Antimicrobial Resistance Monitoring System (NARMS) has shown a recent increase in ESC resistance among Salmonella Heidelberg isolated from food animals at slaughter, retail meat, and humans. ESC resistance among Salmonella in the United States is usually mediated by a plasmid-encoded bla(CMY) ß-lactamase. In 2009, we identified 47 ESC-resistant bla(CMY)-positive Heidelberg isolates from humans (n=18), food animals at slaughter (n=16), and retail meats (n=13) associated with a spike in the prevalence of this serovar. Almost 90% (26/29) of the animal and meat isolates were isolated from chicken carcasses or retail chicken meat. We screened NARMS isolates for the presence of bla(CMY), determined whether the gene was plasmid-encoded, examined pulsed-field gel electrophoresis patterns to assess the genetic diversities of the isolates, and categorized the bla(CMY) plasmids by plasmid incompatibility groups and plasmid multi-locus sequence typing (pMLST). All 47 bla(CMY) genes were found to be plasmid encoded. Incompatibility/replicon typing demonstrated that 41 were IncI1 plasmids, 40 of which only conferred bla(CMY)-associated resistance. Six were IncA/C plasmids that carried additional resistance genes. pMLST of the IncI1-bla(CMY) plasmids showed that 27 (65.8%) were sequence type (ST) 12, the most common ST among bla(CMY)-IncI1 plasmids from Heidelberg isolated from humans. Ten plasmids had a new ST profile, ST66, a type very similar to ST12. This work showed that the 2009 increase in ESC resistance among Salmonella Heidelberg was caused mainly by the dissemination of bla(CMY) on IncI1 and IncA/C plasmids in a variety of genetic backgrounds, and is likely not the result of clonal expansion.

Salmonella prevalence and antimicrobial susceptibility from the National Animal Health Monitoring System Swine 2000 and 2006 studies.

Concern about Salmonella contamination of food is compounded by fear that antimicrobials traditionally used to combat the infection will become useless due to rising antibiotic resistance. Livestock, in particular swine, often are blamed for illnesses caused by Salmonella and for increasing antibiotic resistance due to use of antibiotics in pigs. As part of the National Animal Health Monitoring System Swine 2000 and 2006 studies, swine fecal samples were cultured for Salmonella. These samples were collected from 123 operations in 17 states in 2000 and from 135 operations in 17 states in 2006. At each operation, 50 and 60 fecal samples were collected from late finisher pig pens in 2000 and 2006, respectively. Salmonella isolates were characterized to determine serogroup and serotype and were tested for susceptibility to a panel of 17 and 15 antimicrobial drugs in 2000 and 2006, respectively. A total of 5,470 and 7,788 samples were cultured for Salmonella in 2000 and 2006, respectively. Overall, 6.2% of the samples and 34.2% of the farms were positive for Salmonella in 2000. In 2006, 7.2% of the samples and 52.6% of the farms were positive. Salmonella Derby, Salmonella Typhimurium var. 5- (formerly Salmonella Typhimurium var. Copenhagen), and Salmonella Agona were the three serotypes most often recovered in both study years. The most common antimicrobial resistance pattern for Salmonella Derby in the two study years was resistance to streptomycin, sulfisoxazole, and tetracycline. Most isolates were resistant to tetracycline, sulfisoxazole, and streptomycin in both study years. The proportion of Salmonella isolates that were susceptible to all antimicrobials (pansusceptible) was 38.1% in 2000 and 20.4% in 2006. The proportion of Salmonella isolates that were resistant to three or more antimicrobials (multidrug resistant) was similar in 2000 and in 2006 (52.8 and 57.7%, respectively).

Clostridium difficile from healthy food animals: optimized isolation and prevalence.

Two isolation methods were compared for isolation of Clostridium difficile from food animal feces. The single alcohol shock method (SS) used selective enrichment in cycloserine-cefoxitin fructose broth supplemented with 0.1% sodium taurocholate, followed by alcohol shock and isolation on tryptic soy agar supplemented with 5% sheep blood, and cycloserine-cefoxitin fructose agar. The double alcohol shock method (DS) used alcohol shock prior to and after selective enrichment in cycloserine-cefoxitin fructose broth supplemented with 0.1% sodium taurocholate, followed by isolation on tryptic soy agar supplemented with 5% sheep blood and cycloserine-cefoxitin fructose agar. A total of 55 (15.9%, n = 345) swine fecal samples, 32 (2.4%, n = 1,325) dairy cattle fecal samples, and 188 (6.3%, n = 2,965) beef cattle fecal samples were positive for C. difficile by either method. However, the DS was significantly better than the SS for the recovery of C. difficile from swine feces, while the SS was significantly better than the DS for the recovery of C. difficile from beef cattle feces. There was no significant difference between methods for the recovery of C. difficile from dairy cattle feces. This study suggests that food animals might harbor C. difficile and it provides critical information that isolation methods might not have universal application across animal species.

Anatomical distribution and genetic relatedness of antimicrobial-resistant Escherichia coli from healthy companion animals.

AIMS: Escherichia coli have been targeted for studying antimicrobial resistance in companion animals because of opportunistic infections and as a surrogate for resistance patterns in zoonotic organisms. The aim of our study is to examine antimicrobial resistance in E. coli isolated from various anatomical sites on healthy dogs and cats and identify genetic relatedness. METHODS AND RESULTS: From May to August, 2007, healthy companion animals (155 dogs and 121 cats) from three veterinary clinics in the Athens, GA, USA, were sampled. Escherichia coli was isolated from swabs of nasal, oral, rectal, abdomen and hindquarter areas. Antimicrobial susceptibility testing against 16 antimicrobials was performed using broth microdilution with the Sensititre™ system. Clonal types were determined by a standardized pulsed-field gel electrophoresis protocol. Although rectal swabs yielded the most E. coli (165/317; 52%) from dogs and cats, the organism was distributed evenly among the other body sites sampled. Escherichia coli isolates from both dogs and cats exhibited resistance to all antimicrobials tested with the exception of amikacin, cephalothin and kanamycin. Resistance to ampicillin was the most prevalent resistance phenotype detected (dogs, 33/199; 17%; and cats, 27/118; 23%). Among the resistant isolates, 21 resistance patterns were observed, where 18 patterns represented multidrug resistance (MDR; resistance ≥ 2 antimicrobial classes). Also among the resistant isolates, 33 unique clonal types were detected, where each clonal type contained isolates from various sampling sites. Similar resistance phenotypes were exhibited among clonal types, and three clonal types were from both dogs and cats. CONCLUSIONS: Healthy companion animals can harbour antimicrobial-resistant E. coli on body sites that routinely come in contact with human handlers. SIGNIFICANCE AND IMPACT OF THE STUDY: This study is the first report that demonstrates a diverse antimicrobial-resistant E. coli population distributed over various sites of a companion animal's body, thereby suggesting potential transfer of resistant microflora to human hosts during contact.

Prevalence, species distribution and antimicrobial resistance of enterococci isolated from US dairy cattle.

AIMS: To estimate prevalence and antimicrobial susceptibility of enterococci in faeces collected in 2007 from U.S. dairy cattle. METHODS AND RESULTS: A total of 718 faecal samples from 122 dairy cattle operations from 17 US States were collected and cultured for the presence of enterococci. One hundred and eighteen of the 122 operations (96·7%) had at least one dairy cow positive for enterococci and 88·7% (637 of 718) of the faecal samples were positive. At least ten different enterococcal species were found on the dairy operations and 90·7% (107 of 118) of the operations were positive for Enterococcus hirae followed by E. faecalis (40·7%; 48 of 118) and E. faecium (39%; 46 of 118). The highest percentage of resistant isolates were to lincomycin (92·3%; 587 of 636), flavomycin (71·9%; 457 of 636) and tetracycline (24·5%; 156 of 636). Multi-drug resistance (MDR) (resistance ≥ 2 antimicrobials) was observed to as many as seven antimicrobials regardless of class. CONCLUSION: In contrast to previous studies, faecal shedding of enterococci in dairy cattle occurred in almost 90% of cows sampled and represented a variety of enterococcal species. SIGNIFICANCE AND IMPACT OF STUDY: Although this study demonstrated a high prevalence of antimicrobial-resistant enterococci from dairy cattle faeces in the United States, the contribution of dairy cattle as a source of antimicrobial-resistant enterococci that can be transmitted to humans remains unclear.

Comparison of neck skin excision and whole carcass rinse sampling methods for microbiological evaluation of broiler carcasses before and after immersion chilling.

Sampling protocols for detecting Salmonella on poultry differ among various countries. In the United States, the U.S. Department of Agriculture Food Safety and Inspection Service dictates that whole broiler carcasses should be rinsed with 400 ml of 1% buffered peptone water, whereas in the European Union 25-g samples composed of neck skin from three carcasses are evaluated. The purpose of this study was to evaluate a whole carcass rinse (WCR) and a neck skin excision (NS) procedure for Salmonella and Escherichia coli isolation from the same broiler carcass. Carcasses were obtained from three broiler processing plants. The skin around the neck area was aseptically removed and bagged separately from the carcass, and microbiological analysis was performed. The corresponding carcass was bagged and a WCR sample was evaluated. No significant difference (alpha

Mechanisms of antimicrobial resistance and genetic relatedness among enterococci isolated from dogs and cats in the United States.

AIMS: In this study, mechanisms of antimicrobial resistance and genetic relatedness among resistant enterococci from dogs and cats in the United States were determined. METHODS AND RESULTS: Enterococci resistant to chloramphenicol, ciprofloxacin, erythromycin, gentamicin, kanamycin, streptomycin, lincomycin, quinupristin/dalfopristin and tetracycline were screened for the presence of 15 antimicrobial resistance genes. Five tetracycline resistance genes [tet(M), tet(O), tet(L), tet(S) and tet(U)] were detected with tet(M) accounting for approx. 60% (130/216) of tetracycline resistance; erm(B) was also widely distributed among 96% (43/45) of the erythromycin-resistant enterococci. Five aminoglycoside resistance genes were also detected among the kanamycin-resistant isolates with the majority of isolates (25/36; 69%) containing aph(3')-IIIa. The bifunctional aminoglycoside resistance gene, aac(6')-Ie-aph(2'')-Ia, was detected in gentamicin-resistant isolates and ant(6)-Ia in streptomycin-resistant isolates. The most common gene combination among enterococci from dogs (n = 11) was erm(B), aac(6')-Ie-aph(2'')-Ia, aph(3')-IIIa, tet(M), while tet(O), tet(L) were most common among cats (n = 18). Using pulsed-field gel electrophoresis (PFGE), isolates clustered according to enterococcal species, source and antimicrobial gene content and indistinguishable patterns were observed for some isolates from dogs and cats. CONCLUSION: Enterococci from dogs and cats may be a source of antimicrobial resistance genes. SIGNIFICANCE AND IMPACT OF THE STUDY: Dogs and cats may act as reservoirs of antimicrobial resistance genes that can be transferred from pets to people. Although host-specific ecovars of enterococcal species have been described, identical PFGE patterns suggest that enterococcal strains may be exchanged between these two animal species.

Campylobacter species occurrence within internal organs and tissues of commercial caged Leghorn laying hens.

Campylobacter spp. are frequently present in the intestinal tract and internal tissues of broiler breeder and broiler chickens. Campylobacter spp. ecology in commercial Leghorn laying hens has not been extensively studied. The objectives of the current study were to determine 1) Campylobacter spp. presence in the reproductive tract, lymphoid organs, liver-gallbladder, and ceca of commercial Leghorn laying hens; 2) species of Campylobacter present; and 3) antimicrobial resistance pattern of Campylobacter isolates. In study 1, three flocks ranging from 94 to 105 wk of age were sampled from a commercial laying complex. In study 2, two flocks, 82 and 84 wk of age, were sampled from a separate complex. Hens were killed, defeathered, aseptically necropsied, and the spleen, liver-gallbladder, ovarian follicles, and upper (infundibulum, magnum, and isthmus) and lower (shell gland and vagina) reproductive tracts were aseptically removed before the ceca. Samples were packed on ice and transported to the laboratory for evaluation. For speciation, a standard BAX real-time PCR method was used while susceptibility testing was performed using US National Antimicrobial Resistance Monitoring System (NARMS) standards and recommended quality control organisms. Isolates were examined for susceptibility using a semi-automated testing system (Sensititer) to the following 9 antimicrobials: azithromycin, clindamycin, ciprofloxacin, erythromycin, florfenicol, gentamicin, nalidixic acid, telithromycin, and tetracycline. In study 1, the isolation rate was 13, 67, 53, 3, 13, and 57% from the ovarian follicles, lower reproductive tract, upper reproductive tract, spleen, liver-gallbladder, and ceca, respectively. In study 2, the isolation rate was 17, 43, 33, 20, 17, and 73% from the ovarian follicles, lower reproductive tract, upper reproductive tract, spleen, liver-gallbladder, and ceca, respectively. Overall, 50% of isolates were Campylobacter jejuni, 49% Campylobacter coli, and 1% Campylobacter lari. In study 1, all of the isolates were pan-susceptible. In study 2, thirty-seven percent of the isolates were resistant to tetracycline. Commercial table egg laying hens housed in colony cages on wire floors had diverse Campylobacter spp. recovered from different tissues and these isolates were not resistant to a broad range of antimicrobials.

Prevalence, serotype, and antimicrobial resistance of Salmonella on broiler carcasses postpick and postchill in 20 U.S. processing plants.

The objective of this study was to measure the effect of broiler processing on the prevalence, serotype, and antimicrobial resistance profiles of salmonellae. Twenty U.S. commercial processing plants representing eight integrators in 13 states were included in the survey. In each of four replications, 10 carcasses from one flock were collected at rehang and 10 more carcasses were collected at postchill; each carcass was sampled by whole-carcass rinse. Salmonella organisms were isolated from carcass rinses by standard cultural techniques, serotypes were determined, and the resistance to 15 antimicrobials was measured. Overall, Salmonella was detected on 72% of carcasses at rehang (ranging from 35 to 97%) and on 20% of carcasses postchill (ranging from 2.5 to 60%). In every instance, a significant (P < 0.05) decrease in Salmonella prevalence was noted between rehang and postchill. The four most common serotypes, accounting for 64% of all Salmonella isolates, were Kentucky, Heidelberg, Typhimurium, and Typhimurium var. 5-; most isolates of Kentucky (52%), Heidelberg (79%), and Typhimurium (54%) serotypes were susceptible to all antimicrobial drugs tested. However, only 15% of the Typhimurium var. 5- isolates were pansusceptible; more than one-half of the isolates of this serotype were resistant to three or more drugs. No isolate of any serotype exhibited resistance to amikacin, ceftriaxone, ciprofloxacin, or trimethoprim-sulfamethoxazole. These data demonstrate that although processing lessens carcass contamination with Salmonella, antimicrobial-resistant isolates may still be present.