Diversity of Shiga toxin-producing Escherichia coli in sheep flocks of Paraná State,southern Brazil

Sheep constitute an important source of zoonotic pathogens as Shiga toxin-producing Escherichia coli (STEC). In this study, the prevalence, serotypes and virulence profiles of STEC were investigated among 130 healthy sheep from small and medium farms in southern Brazil. STEC was isolated from 65 (50%) of the tested animals and detected in all flocks. A total of 70 STEC isolates were characterized, and belonged to 23 different O:H serotypes, many of which associated with human disease, including hemolytic-uremic syndrome (HUS). Among the serotypes identified, O76:H19 and O65:H– were the most common, and O75:H14 and O169:H7 have not been previously reported in STEC strains. Most of the STEC isolates harbored only stx1, whereas the Stx2b subtype was the most common among those carrying stx2. Enterohemolysin (ehxA) and intimin (eae) genes were detected in 61 (87.1%) and four (5.7%) isolates, respectively. Genes encoding putative adhesins (saa, iha, lpf_O113) and toxins (subAB and cdtV) were also observed. The majority of the isolates displayed virulence features related to pathogenesis of STEC, such as adherence to epithelial cells, high cytotoxicity and enterohemolytic activity. Ovine STEC isolates belonged mostly to phylogenetic group B1. PFGE revealed particular clones distributed in some farms, as well as variations in the degree of genetic similarity within serotypes examined. In conclusion, STEC are widely distributed in southern Brazilian sheep, and belonged mainly to serotypes that are not commonly reported in other regions, such as O76:H19 and O65:H–. A geographical variation in the distribution of STEC serotypes seems to occur in sheep.     

Recovery and Genetic Diversity of Escherichia coli Isolates from Deep Litter,Shallow Litter,and Surgical Shoe Covers

Litter is a common source of infectious agents in poultry production environments. Typical sampling methods only examine bacteria on the surface; however, bacteria recovered with these methods may not be representative of the population in the litter. To test this hypothesis, both shallow (top 2 in.) and deep (bottom 2 in.) litter samples were taken and compared with a surface sampling method (i.e., surgical shoe covers). Escherichia coli isolates were recovered from all 3 sample types and examined using an automated ribotyping system to determine the genetic diversity of the isolates. Twenty-six unique E. coli strains were recovered from the 3 different sampling methods. There was no correlation among strains between visit age, house, flock, or farm and ribogroups. Based upon the patterns of E. coli recovery among the different sample types, our results suggest that surface sampling methods are equally capable of recovering common isolates from the litter. Surgical shoe covers were easy to use, provided the same core population of isolates, and were comparable to shallow litter in the number of strains recovered.     

Antibiotic Resistance in Salmonella Isolated from Tegus (Tupinambis spp.)

In recent years, an increase in human clinical cases of reptile-associated salmonellosis has been identified, and it has been attributed to the increased popularity of these animals as pets. Limited information is available regarding the distribution of Salmonella spp. serotypes in different reptile species and the antimicrobial resistance patterns of Salmonella spp. isolated from pet reptiles. This article describes the prevalence of Salmonella spp., distribution of serotypes, and antibiotic susceptibility patterns from isolates cultured from cloacal swabs obtained from 14 tegu lizards (Tupinambis spp.). Eighteen strains of Salmonella belonging to different serotypes were obtained from the 14 tegu lizards. Of the 18 Salmonella spp. isolates, 8 (44.4%) were from Salmonella subspecies I, with a majority of isolates belonging to the Eastbourne serotype (3 strains), Nottingham serotype (2 strains), and Brancaster serotype (2 strains), and only 1 belonging to the Apapa serotype. Less common serotypes were detected in 5 isolates, including 2 each belonging to Salmonella subspecies II and IIIb, respectively, and 1 to Salmonella subspecies IIIa. The serotype of 5 other Salmonella isolates could not be determined. All 18 isolates were resistant to at least 6 of the antimicrobial drugs tested. These results confirm the potential zoonotic risk from handling reptiles, suggesting that measures to educate the reptile-owning public are necessary.     

Prevalence and antimicrobial susceptibility of Salmonella isolates in Pekin ducks from South Korea

An investigation was carried out to determine the prevalence and antibiotic resistance of Salmonella serotypes at South Korean duck farms. A total of 7119 samples collected from 72 duck farms in five provinces were examined from 2011 to 2012. The overall prevalence of Salmonella serotypes was 43.4% (69/159) in duck flocks from 65.2% (47/72) of the duck farms. Eighty-five strains were isolated from 69 duck flocks. Three serotypes of Salmonella enterica were identified such as S. Typhimurium (39/85), S. Enteritidis (44/85), and S. London (2/85). The prevalence of Salmonella infection decreased significantly in 3-week-old ducks compared to that in 1-week-old ducks (P < 0.05). All isolates except one were resistant to at least one antimicrobial and 27% of the isolates were resistant to 5–16 antimicrobials. Our findings provide baseline information on the degree of Salmonella infection and distribution of Salmonella serotypes in ducks and indicate that ducks should be considered an important source of foodborne pathogens.     

Diversity of Mannheimia haemolytica and Pasteurella trehalosi Serotypes from Apparently Healthy Sheep and Abattoir Specimens in the Highlands of Wollo,North East Ethiopia

The prevalence and serotypic diversity of Mannheimia [Pasteurella] haemolytica and Pasteurella trehalosi from nasal swabs, sera and abattoir specimens from sheep in the highlands of Wollo, North East Ethiopia was investigated. Prevalence rates of 83% and 75% of these microorganisms were found in the serum samples and nasal swabs, respectively, from apparently healthy sheep. In a local abattoir, 205 lungs were investigated, 34% of which showed pneumonia, from which samples were collected from 51 lungs and the same number of corresponding tonsils. Mannheimia and Pasteurella species were isolated from 59% of these pneumonic lungs and 69% of the respective tonsils. M. haemolytica serotypes accounted for 41 (59%) and P. trehalosi for 11 (32%) of the isolates from the abattoir specimens. The majority (67%) of isolates from nasal swabs were P. trehalosi, M. haemolytica being isolated f rom 4 (13%) of the swabs. M. glucosida was isolated only from the tonsils. The predominant serotypes of the isolates from both the nasal swabs and the abattoir specimens were M. haemolytica A1 (17%) and P. trehalosi T4 (16%) and T3 (13%). P. trehalosi T15 was less commonly encountered, while M. haemolytica A9 and A13 were not isolated. Studies on sera from 100 sheep indicated that antibodies against M. haemolytica serotype A1 (14%) were most common, followed by A5 and A8 (each 10%) and A9 and P. trehalosi T3 (each 9%) and T4 (8%). Antibodies against M. glucosida or serotype A11 occurred in 2% of the sera. Multiple serotypes were common in all types of samples. The importance of including in vaccines the most prevalent serotypes involved in the pneumonia of sheep in the area is discussed.     

Effect of subtherapeutic antimicrobials on genetic diversity of Enterococcus faecium from chickens

The effect of growth promotants (bacitracin, virginiamycin, and flavomycin) on the genetic population of Enterococcusfaecium isolated from a commercially integrated poultry farm was examined. A total of 551 E. faecium were isolated from chick boxliners (n=16), litter (n=334), feed (n=67), and carcass rinse (n=134) samples from four chicken houses. Two houses on the farm were control houses and did not use any antimicrobials while two other houses on the farm used flavomycin, virginiamycin, and bacitracin during six different chicken grow outs. BOX-PCR and pulsed-field gel electrophoresis (PFGE) results indicated that E. faecium strains had a high degree of genetic diversity as overall clustering was independent of source, house, or grow out. Similarity of > or =60% for the majority of BOX-PCR genogroups and > or =80% for the majority of PFGE genogroups was observed for a subset of carcass rinse samples (n=45) examined. Seventy-nine percent (19/24) of isolates in BOX-PCR genogroup 2 also clustered in PFGE genogroup 2, although no association between the isolates and house or grow out was observed. These results suggest that E. faecium from chicken are genetically diverse and that growth-promoting antimicrobials do not affect the genetic population of E. faecium.     

Prevalence and antimicrobial susceptibility of Salmonella spp. in small Indian mongooses (Herpestes auropunctatus) in Grenada,West Indies

Intestinal samples from 156 small Indian mongooses (Herpestes auropunctatus) collected island-wide in Grenada from April 2011 to March 2013 were examined for the presence of Salmonella enterica spp. Nineteen (12%) mongooses were culture-positive for S. enterica spp. of which five serotypes were identified. Salmonella javiana and S. Montevideo were the most commonly isolated serotypes. The other serotypes isolated were S. Rubislaw, S. Panama and S. Arechavaleta. All isolates were susceptible to amoxicillin–clavulanic acid, ampicillin, cefotaxime, ceftazidime, ciprofloxacin, enrofloxacin, gentamicin, nalidixic acid, imipenem and trimethoprim–sulfamethoxazole. One isolate (S. Montevideo) showed resistance to tetracycline and intermediate resistance to streptomycin. The five isolated Salmonella serotypes are potential human pathogens suggesting that the mongoose may play a role in the epidemiology of human salmonellosis in Grenada.     

Distribution of oxytetracycline resistance genes in E. coli isolated from pigeon faecal samples

Tetracycline- resistant bacteria have emerged due to the selective pressure of antimicrobial use. The aim of this study was to determine the prevalence of oxytetracycline resistance of Escherichia coli from pigeon faecal samples. All strains were examined for the presence and types of oxytetracycline resistance determinants using disc diffusion and polymerase chain reaction methods. Of 50 faecal E. coli isolates, 30 (60%) were resistant to oxytetracycline. Polymerase chain reaction analyses indicated that the most common genes found in these isolates were tetB (43.3%) and tetA (30%). Only 10% and 3.3% of the isolates contained otrA and otrB, respectively. In conclusion, our findings suggest that oxytetracycline-resistant strains of E. coli are disseminated in pigeons.     

Reptile‐Associated Salmonellosis in Minnesota, 1996–2011

Reptile‐associated salmonellosis (RAS) occurs when Salmonella is transmitted from a reptile to a human. This study describes the epidemiology of RAS in Minnesota during 1996–2011. All Minnesotans with confirmed Salmonella infections are reported to the Minnesota Department of Health (MDH). Case patients are interviewed about illness characteristics and risk factors, including foods eaten, drinking and recreational water exposures, contact with ill people, and animal contact. Willing RAS case patients can submit stool from the reptile for culture. Serotype and pulsed‐field gel electrophoresis (PFGE) subtype of Salmonella isolates from reptiles and case patients are compared. Of 8389 sporadic (not associated with an outbreak) non‐typhoidal salmonellosis case patients in Minnesotans during 1996–2011, 290 (3.5%) reported reptile exposure. The median age of case patients with reptile exposure was 11 years, 31% were under the age of 5 years and 67% were under the age of 20 years; 50% were female. The median illness duration was 8 days; 23% required hospitalization. The most commonly reported reptile exposures were lizard (47%), snake (20%), turtle (19%) and a combination of reptile types (14%). Eighty‐four per cent of isolates from case patients who reported reptile exposure were Salmonella enterica subspecies I. The three most common serotypes were Typhimurium (15%), Enteritidis (7%) and subspecies IV serotypes (7%). Of 60 reptiles testing positive for Salmonella, 36 (60%) yielded the same Salmonella serotype as the human isolate. Twenty‐six of 27 reptile isolates that were subtyped by PFGE were indistinguishable from the human isolate. Of these, 88% were subspecies I; the most common serotypes were Enteritidis (12%), Typhimurium (8%), and Bareilly (8%). RAS accounts for approximately 3.5% of salmonellosis cases in Minnesota, primarily affecting children. The majority of isolates from case patients and reptiles belonged to Salmonella subspecies I, suggesting that reptiles are a source of human infection with serotypes not traditionally considered to be reptile‐associated.     

Serological characterization and antimicrobial susceptibility of Streptococcus suis isolates from diagnostic samples in Denmark during 1995 and 1996

At the Danish Veterinary Laboratory Streptococcus suis infections in pigs were diagnosed in 114 cases in 1995 and in 151 cases in 1996. Isolates were serotyped using specific antisera against type 1 through 28 and a total of 67 cases from 1995 and 113 cases in 1996 were tested for resistance to 11 antimicrobial agents. The majority of cases were lung diseases (57%), followed by septicaemia (16%), meningitis (15%) and endocarditis (8%). Almost 96% of the isolates could be typed using the 28 antisera. The most common serotype was serotype 2 (29%), followed by serotype 7 (17%), and serotypes 3, 4 and 8 (9–10%). The remaining serotypes were observed in frequencies of less than 5%. Serotype 7 was more commonly isolated from septicaemia than the other serotypes. Serotype 2 was more commonly isolated from pigs older than 4 weeks compared to the other serotypes. Most isolates were susceptible to amoxycillin+clavulanate, ampicillin, ceftiofur, enrofloxacin, penicillin, spectinomycin, tiamulin and trimethoprim+sulphadiazine. A high frequency (>30%) of resistance to tetracycline was observed. Among isolates of serotype 2, 9.7% were resistant to lincomycin and 12.9% to spiramycin. Among other serotypes 56.8% were resistant to lincomycin and spiramycin. The differences in susceptibility between isolates of serotype 2 and the other serotypes were statistically significant. Compared to a previous Danish study the distribution of serotypes of S. suis causing infections among pigs in Denmark has changed during the last 15 years.     

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.     

A National Surveillance of Shiga Toxin‐Producing Escherichia coli in Food‐Producing Animals in Japan

To assess the public health risk, the prevalence and anti‐microbial resistance of Shiga toxin‐producing Escherichia coli (STEC) among food‐producing animals were studied throughout Japan. Faecal samples were collected from healthy animals of 272 cattle, 179 pigs, and 158 broilers on 596 farms in all 47 Japanese prefectures. STEC were isolated from 62 (23%) cattle and 32 (14%) pig samples but from no chicken samples. Of the bovine isolates, 19 belonged to serotypes frequently implicated in human disease (O157:H7/non‐motile (NM)/H not typeable, O26:NM/H11/H21/H not typeable, O113:H21, and O145:NM). The eae genes were observed in 37% of bovine isolates; among them one O145:NM and all four O157 isolates possessed eae‐γ1, and one O145:NM, one O103:H11, and all five O26 isolates possessed eae‐β1 gene. Among the swine isolates, stx2e were dominant, and serotypes frequently implicated in human diseases or eae‐positive isolates were not observed. Bovine isolates showed less anti‐microbial resistance, but six isolates of 26:NM/H11 and O145:NM were multi‐resistant and may need careful monitoring. Swine isolates showed various resistance patterns; chloramphenicol resistance patterns were more common than in bovine isolates. This first national study of STEC in the Japanese veterinary field should aid our understanding of Japan's STEC status.     

Evaluation of Salmonella serotype distributions from commercial broiler hatcheries and grower houses

By conventional trayliner (hatcheries) and drag swab assembly (broiler houses) culture methods, the isolation distribution of Salmonella serotypes from five commercial broiler hatcheries (three sample times) and 13 broiler farms (eight sample times) was evaluated. A total of 11 different Salmonella serotypes were isolated from hatcheries, with Salmonella heidelberg (9/30) and Salmonella kentucky (6/30) accounting for 50% of the total isolations. Of 700 chick paperpad trayliners sampled, regardless of lot (breeder flock source) or hatchery, 12% were positive for Salmonella. When 10 individual trayliners were cultured from individual lots (same breeder flock source), Salmonella was detected in 24/57 lots (42%). Multiple serotypes were simultaneously isolated from the same lot on three occasions (6%). Of the 21 lots that were serially sampled, the Salmonella serotype detected was different within lots eight times (38%) on at least one occasion of two or more sampling times. Of the 196 individual broiler houses sampled, 44 were positive for Salmonella (42%). Twelve different serotypes were isolated from broiler houses during this study. The serotypes isolated most frequently were S. heidelberg (34/94) and S. kentucky (22/94). These two serotypes accounted for 59.6% (56/94) of the total broiler house isolations. Of the 38 houses that were serially sampled, two or more serotypes were detected in the same broiler house on 20 occasions (53%). Of the 38 serially sampled houses (four or more times), a consistent Salmonella serotype was detected in five houses (13%). In only 5 of the 38 (13%) serially sampled houses did we fail to detect Salmonella on four or more samplings. No significant difference in Salmonella isolation frequency was observed between poultry houses using new or used litter. These data support previous findings indicating that paratyphoid Salmonella serotypes are prevalent in some broiler hatcheries and houses. Further, the observation of multiple serotypes simultaneously and serially isolated from the same breeder hatchery lots suggests that breeder flocks may be infected with more than one serotype, possibly providing a source for multiple serotype infections in progeny grower flocks.     

Staphylococcal and other Bacterial Species Associated with Intramammary Infections in Danish Dairy Herds

Four thousand six hundred forty– five quarter milk samples from 1179 cows from 20 commercial dairy herds were examined in order to determine the prevalence of bacterial species. A total of 859 isolates from 839 (18.1%) culture positive samples could be assigned to 34 different species and subspecies. Diagnostics of staphylococcal species was based on conventional procedures able to differentiate between all 36 species and subspecies presently acknowledged. Staphylococcus aureus was found in 10.2% of the samples and was the most common species isolated. Streptococcus dysgalactiae (1.6%) and Streptococcus uberis (1.4%) were the second and third most common species isolated. Seventeen different coagulase negative staphylococcal species (CNS) were found in 4.1% of the samples. The most frequently isolated CNS were S. epidermidis (1.3%), S. chromogenes (1.0%) and S. simulans (0.7%). Isolates of S. aureus were phage typed, and isolates of S. epidermidis were investigated by phage typing, antibiogram typing, and biotyping. A total of 378 (79.9%) isolates of S. aureus could be typed by phages, assigning them to 18 different phage types. However, 6 phage types accounted for 92.1% of the typable isolates. One to 2 phage types predominated within each herd. Eleven (18%) isolates of S. epidermidis could be typed by phages, assigning the isolates to 3 different types. Biotyping of S. epidermidis produced a total of 8 different types, the most common accounting for 29.5% of the isolates. A total of 6 different antibiogram types were observed among all isolates of S. epidermidis. Resistance towards penicillin (36.1%), tetracycline (9.8%) and streptomycin (9.8%), were recorded in the isolates of S. epidermidis. However, 35 (57.4%) of the isolates were susceptible to all 12 antibiotics tested.     

Prevalence of capsular serotypes among Staphylococcus aureus isolates from cows with mastitis in the United States

Development of an appropriate Staphylococcus aureus vaccine for bovine mastitis has eluded researchers for decades. The ability of S. aureus to form a protective exopolysaccharide capsule has posed a major obstacle because of the multiple serotypes and the poor immune response elicited by exopolysaccharides. This study characterized S. aureus serotypes isolated from cases of bovine mastitis obtained from veterinary diagnostic laboratories that service 44% of the dairy cattle in the United States. Major milk producing areas of the northeast, north central, Pacific coast and southwest were proportionately represented. Sub-samples of mastitic milk that contained S. aureus were frozen and sent to our laboratory for strain serotyping. The only other regional serotyping of S. aureus from bovine mastitis to date was done in France. The primary serotypes found were types 5 (51%) and 8 (18%) and 31% were non-typeable. In the current study, serotype 5 accounted for 18% of the isolates and serotype 8 for 23%. More importantly 59% of the isolates were not typeable with either type 5 or 8 antisera. These data indicate that S. aureus vaccines employing serotypes 5 and 8 would only be marginally effective in the United States. These data also suggest that development of a S. aureus vaccine for bovine mastitis should take into account regional variation in S. aureus serotypes.     

Serotypes of Salmonella Isolated from Faeces of Patients with Acute Diarrhoea in Gwangju Area,Korea, During 2000–2009

The purpose of this study was to determine the changing pattern of Salmonella serotypes causing acute diarrhoea in humans in Gwangju area, Korea, during 2000–2009. A total of 596 Salmonella isolated from culture of 29 896 faecal samples of patients with acute diarrhoea were included in this study. Faecal samples were collected from local hospitals and clinics in Gwangju area during January 2000–December 2009. The mean annual frequency of isolates for the 10 years was 2.0% (range, 0.9–6.0). The isolates were serologically classified into 43 different serotypes. The 10 most common serotypes were Salmonella Enteritidis (47.9%), S. Typhimurium (20.4%), S. Braenderup (3.2%), S. Montevideo (2.9%), S. Paratyphi B (2.9%), S. London (2.3%), S. Bardo (1.7%), S. Virchow (1.7%), S. Infantis (1.5%) and S. Typhi (1.5%), accounting for 86% of all the isolates. Temporal variations were observed in the distribution of different Salmonella serotypes over the years, and only S. Enteritidis and S. Typhimurium were persistent throughout the study period. Although age specificity varied with serotypes, Salmonella was isolated most frequently from children below 5 years of age (179/596, 30.0%). A seasonal trend was apparent, and the highest rates were found in the summer months. This is the first report of the annual frequency of isolation of Salmonella serotypes, and seasonal and age‐specific patterns of salmonellosis in humans in Gwangju area, Korea, over a decade‐long period.     

Genetic diversity of Streptococcus suis serotypes 2 and 1/2 isolates recovered from carrier pigs in closed herds

The aim of this study was to compare, by randomly amplified polymorphic DNA (RAPD), the diversity of Streptococcus suis serotypes 1/2 and 2 isolates recovered at slaughter houses from the tonsils of clinically healthy pigs. The pigs belonged to herds with or without clinical signs of S. suis disease.

Overall, a low diversity was observed among isolates of serotype 1/2. A representative isolate recovered from a diseased animal presented a relatively high similarity (85%), with most isolates recovered from carrier pigs, from herds either with or without clinical signs of S. suis disease. For serotype 2 isolates, a relatively high degree of heterogeneity was observed in the whole population. Two subpopulations were observed for serotype 2 isolates, which arose from herds with clinical signs. Interestingly, the representative isolate coming from the diseased pig was included in a small closed cluster, with 2 isolates recovered from carrier pigs belonging to the same herd. On the other hand, most of the S. suis serotype 2 isolates originating from herds with no history of S. suis disease, were closely related (90% similarity). Furthermore, they presented different RAPD patterns from those originating from animals from the herd presenting S. suis clinical signs due to this serotype. Results suggest that, in the herds studied, clinical manifestations due to serotype 2 are probably related to the virulence of a specific isolate. Conversely, for the herd affected with serotype 1/2, clinical manifestations of the disease were more likely to be the result of inherent herd factors than the virulence of the specific isolate.

     

Clinically relevant multidrug resistant Salmonella enterica in swine and meat handlers at the abattoir

The presence of multidrug resistant (MDR) Salmonella serotypes in slaughtered swine, carcasses, meat and meat handlers is scarcely evaluated. Recently we demonstrated that diverse Salmonella serotypes are frequently present in swine, pork meat and carcasses, and meat handlers at Portuguese abattoirs. Here we have characterized their antibiotic resistance phenotypes and genotypes, helping elucidate the flow of MDR Salmonella in the food chain. Testing 60 Salmonella isolates from different serotypes, the highest frequencies of resistance were observed for tetracycline (T) [70% (n = 42/60), tet(A)/tet(B)/tet(G)], streptomycin (S) [63% (n = 38/60), aadA2/strA/strB], sulfamethoxazole (Sul) [62% (n = 37/60), sul1/sul2/sul3] and ampicillin (A) [57% (n = 34/60), bla_PSE-1/bla_TEM]. Thirty-seven percent (n = 22/60) carried class 1 integrons and multidrug resistance was frequently observed (63% n = 38/60), including those serotypes common to human infections [S. Typhimurium 78% n = 25/32; S. 4,[5],12:i:- 67% n = 2/3; S. Rissen 75% (n = 3/4); S. London 67% n = 2/3; S. Derby 55%; n = 6/11)]. The emergent S. 4,[5],12:i:- isolates were mostly characterized by ASSuT phenotype [bla_TEM/strA-strB/sul2/tet(B)], typical of the European clone, while for the first time the ST phenotype [strA-strB-tet(A)-tet(B)] was also observed. Moreover, we report a first finding of a MDR phenotype in S. London [ANSSuT; bla_TEM-strA-strB-sul2-tet(A)]. Our findings suggest that the abattoir environment and the slaughter operations seem not only to harbor MDR serotypes that originated in the pig reservoir, but also propagate them through cross-contamination processes, involving meat handlers. The present study suggests a probable relationship between swine and human salmonellosis throughout the food chain, which is of interest for epidemiological, animal health and public health purposes.     

Characterization of Clostridium perfringens in the feces of adult horses and foals with acute enterocolitis

Up to 60% of cases of equine colitis have no known cause. To improve understanding of the causes of acute colitis in horses, we hypothesized that Clostridium perfringens producing enterotoxin (CPE) and/or beta2 toxin (CPB2) are common and important causes of severe colitis in horses and/or that C. perfringens producing an as-yet-undescribed cytotoxin may also cause colitis in horses. Fecal samples from 55 horses (43 adults, 12 foals) with clinical evidence of colitis were evaluated by culture for the presence of Clostridium difficile, C. perfringens, and Salmonella. Feces were also examined by enzyme-linked immunosorbent assay (ELISA) for C. difficile A/B toxins and C. perfringens alpha toxin (CPA), beta2 toxin (CPB2), and enterotoxin (CPE). Five C. perfringens isolates per sample were genotyped for the following genes: cpa, cpb, cpb2 consensus, cpb2 atypical, cpe (enterotoxin), etx (epsilon toxin), itx (iota toxin), netB (necrotic enteritis toxin B), and tpeL (large C. perfringens cytotoxin). The supernatants of these isolates were also evaluated for toxicity for an equine cell line. All fecal samples were negative for Salmonella. Clostridium perfringens and C. difficile were isolated from 40% and 5.4% of samples, respectively. All fecal samples were negative for CPE. Clostridium perfringens CPA and CPB2 toxins were detected in 14.5% and 7.2% of fecal samples, respectively, all of which were culture-positive for C. perfringens. No isolates were cpe, etx, netB, or tpeL gene-positive. Atypical cpb2 and consensus cpb2 genes were identified in 15 (13.6%) and 4 (3.6%) of 110 isolates, respectively. All equine C. perfringens isolates showed far milder cytotoxicity effects than a CPB-producing positive control, although cpb2-positive isolates were slightly but significantly more cytotoxic than negative isolates. Based on this studied population, we were unable to confirm our hypothesis that CPE and CPB2-producing C. perfringens are common in horses with colitis in Ontario and we failed to identify cytotoxic activity in vitro in the type A isolates recovered.     

Comparison of Salmonella isolation rates in different types of egg-layer hen houses in Chiba, Japan

This study investigated the differences in Salmonella isolation rates in environmental samples taken from several types of hen houses in Chiba, Japan. In addition, for the detailed epidemiologic survey, environmental samples, hens, and rodents were collected from Salmonella-contaminated windowless houses on three farms. As a result, Salmonella was isolated from four (80%) of five farms with windowless hen houses; Salmonella enteritidis was isolated from a single windowless house. In contrast, only one serotype of Salmonella was isolated from 1 (6.7%) of 15 farms with open hen houses. In the S. enteritidis-contaminated windowless hen house, the isolation rates of S. enteritidis as compared with the other serotypes were 90.9% of environments, 94.1% of hens, and 86.4% of roof rats (Rattus rattus) that resided in the environments. In reference to the phage type (PT) of these isolates, PT1 was detected in environments and roof rats, and PT9 was detected in both these samples and in hens. Thus, the Salmonella isolation rate in hen houses seems to be associated with whether the premises are windowless or open. Moreover, roof rats appear to be the most important vectors in the spread of S. enteritidis in the windowless hen house because the S. enteritidis PTs coincide with each other.