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Consumption of shell eggs has been associated with Salmonella Enteritidis (SE) infections in humans in the United States. Because of this, the Pennsylvania Egg Quality Assurance Program (PEQAP) was developed and implemented in 1994. The PEQAP involves periodic flock testing and management practices to minimize SE contamination of shell eggs. Subsequently, the U.S. Food and Drug Administration (FDA) introduced a mandatory federal program in 2010 and 2012 for shell egg producers modeled closely after PEQAP to reduce the incidence and prevalence of SE during production, storage, and transport nationwide. In this study, a retrospective epidemiologic analysis was conducted by characterizing SE isolated from commercial layer environment samples and shell eggs submitted to the Animal Diagnostic Laboratory at The Pennsylvania State University using phage typing and pulsed-field gel electrophoresis (PFGE). The objective of this study was to determine the relatedness of SE isolates from hen house environments and shell eggs and to optimize the existing protocols of egg quality assurance programs by identifying the best layer-house environmental sampling time points in order to minimize SE contamination of shell eggs. A total of 94 SE isolates from 65 hen flocks on 35 premises in Pennsylvania recovered during 2007 to 2015 were used in this study. The SE phage type 8 and PFGE fingerprint type JEGX01.0004 most commonly associated with human SE infection was also the predominant type present in layer-house environments and shell eggs. This reconfirms hen house environmental monitoring is an effective method to identify SE-infected flocks. Further, the PEQAP program allowed SE detection of infected flocks earlier than the FDA program as it included an additional environmental test at 29-31 wk of age, enabling the earlier prevention of SE-contaminated shell eggs going to the market. Therefore, it is recommended to refine the sampling time points of the current FDA Egg Rule by adding hen house environmental testing at 29-31 wk of age.

The shedding patterns of Salmonella spp. and MLVA profiles of Salmonella enterica subspecies enterica (I) serotype 1,4,[5],12:i:- were monitored in a 12-month longitudinal observational study of five pig herds to inform management; provide indications of potential hazard load at slaughter; and assist evaluation of MLVA for use by animal and public health practitioners. Twenty pooled faecal samples, stratified by age group, were collected quarterly. When Salmonella was cultured, multiple colonies were characterized by serotyping and where S. Typhimurium-like serovars were confirmed, isolates were further characterized by phage typing and multiple locus variable number tandem repeat analysis (MLVA). Salmonella was detected in 43% of samples. Salmonella 1,4,[5],12:i- was one of several serovars that persisted within the herds and was found among colonies from each production stage. Virtually all Salmonella 1,4,[5],12:i:- isolates were phage type 193, but exhibited 12 different, closely-related MLVA profiles. Salmonella 1,4,[5],12:i:- diversity within herds was low and MLVA profiles were stable indicating colonization throughout the herds and suggesting each farm had an endemic strain. High prevalence of S. 1,4,[5],12:i:- specific shedding among terminal animals indicated high hazard load at slaughter, suggesting that primary production may be an important pathway of S. 1,4,[5],12:i:- into the human food chain, this has implications for on-farm management and the application and targeting control measures and further evidence of the need for effective process control procedures to be in place during slaughter and in pork boning rooms. These findings have implications for animal health and food safety risk mitigation and risk management., (Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.)

The objective of this study was to investigate the hypothesis that some sporadic Salmonella infections in domesticated animals may be associated with Salmonella infections originating from garden birds. Phage type and antimicrobial resistance details of isolates of S. Typhimurium obtained from wild birds were comparable with those from S. Typhimurium infections from domesticated animals or livestock between 2002 and 2010. A small panel of S. Typhimurium isolates (n=37) were characterised by multilocus variable number of tandem repeats analysis (MLVA), pulsed field gel electrophoresis (PFGE) and phage type. The MLVA-PFGE data clustered the strains according to phage type (DT40 or DT56). Within each group there were strains from wild birds and domesticated animals or livestock with MLVA profiles having up to 100% similarity. The results from this study therefore lend support to the hypothesis that Salmonella infections in domesticated animals could be caused by infections carried by wild birds., (Crown Copyright © 2013. Published by Elsevier India Pvt Ltd. All rights reserved.)

A retrospective study analysing Salmonella serovars and antimicrobial resistance data from pigs in Great Britain between 1994 and 2010 was performed. Data were obtained through scanning surveillance and compared with prevalence data from other livestock species as well as human data. During the study period, two serovars, S. Typhimurium and S. Derby, predominated in British pigs. The total number of Salmonella incidents has decreased steadily over the study period, from 360 incidents per year in 1994 to 172 incidents in 2010. S. Typhimurium has been the most common serovar every year, but the relative percentage went down over the past few years. During the same time period, monophasic strains of S. Typhimurium have increased in numbers and were accounting for as much as 25% of incidents in 2010, representing the second most common serovar in British pigs in 2010. Antimicrobial resistance data from S. Typhimurium isolates show that the percentage of isolates which are resistant to six or more antimicrobials has increased from 27.2% in 1994 to 58.3% in 2010 and that the percentage of isolates fully sensitive to the panel of antimicrobials tested was only 3.3% in 2010. S. Typhimurium isolates belonged mainly to phagetypes DT193 and U288 in 2010, and an increase can be seen in the number of isolates belonging to phage type DT193. DT104, which was the predominant phagetype during the 1990s, has gone down considerably and represented less than 5% of all S. Typhimurium isolates in 2010. Monophasic strains of S. Typhimurium belonged mainly to phagetype 193 with less than 20% belonging to phagetype 120. The overall trend shows that the number of Salmonella incidents reported from British pigs has gone down considerably over the 17-year study period, but that the relative percentage of monophasic strains of S. Typhimurium has increased significantly. This trend seems to be in line with observations from other European countries, where an increase of this serovar can be seen both in livestock and in human patients. The increasing percentage of Salmonella-isolates which show multi-resistance is a matter of concern and has to be monitored carefully in order to assess any potential risks this may pose to human patients., (Crown Copyright © 2013. Published by Elsevier B.V. All rights reserved.)

Previous reports have indicated that a small proportion of cattle shedding high levels of Escherichia coli O157:H7 is the main source for transmission of this organism between animals. Cattle achieving a fecal shedding status of 10(4) CFU of E. coli O157:H7/gram or greater are now referred to as supershedders. The aim of this study was to investigate the contribution of E. coli O157:H7 strain type to supershedding and to determine if supershedding was restricted to a specific set of E. coli O157:H7 strains. Fecal swabs (n = 5,086) were collected from cattle at feedlots or during harvest. Supershedders constituted 2.0% of the bovine population tested. Supershedder isolates were characterized by pulsed-field gel electrophoresis (PFGE), phage typing, lineage-specific polymorphism assay (LSPA), Stx-associated bacteriophage insertion (SBI) site determination, and variant analysis of Shiga toxin, tir, and antiterminator Q genes. Isolates representing 52 unique PFGE patterns, 19 phage types, and 12 SBI clusters were obtained from supershedding cattle, indicating that there is no clustering to E. coli O157:H7 genotypes responsible for supershedding. While being isolated directly from cattle, this strain set tended to have higher frequencies of traits associated with human clinical isolates than previously collected bovine isolates with respect to lineage and tir allele, but not for SBI cluster and Q type. We conclude that no exclusive genotype was identified that was common to all supershedder isolates.

Poultry products have consistently been identified as important sources of Salmonella infection in humans, because vertical transfer of infection from breeding hens to progeny is an important aspect of the epidemiology of Salmonella spp. infection within the poultry industry. The aim of this study was to estimate the prevalence of Salmonella contamination in poultry products from 15 different located geographical areas from among the 50 poultry slaughterhouses authorized to operate in Korea and to characterize all the isolates by genotyping, phage typing and antibiotic resistance pattern. Salmonella was isolated from 10 (66.7%) of the first and 5 (33.3%) of the last chilling waters and from 32 (42.7%) carcasses originating from 9 slaughterhouses. The major prevalent serotypes of Salmonella originating from 2 duck slaughterhouses and 13 chicken slaughterhouses tested were S. Typhimurium and S. Enteritidis, respectively. Regarding the characteristics of their antibiotic resistance, 8 of the 11 ampicillin resistant (AmR) isolates carried blaTEM only, two carried blaTEM and blaCTX-M-14 and one carried blaCTX-M-3 and only one AmR isolate with the blaCTX-M-3 β-lactamase gene was an ESBL-producing Salmonella strain. Twenty-seven Salmonella isolates showed nalidixic acid resistance with a mutation at amino acid codon Asp87 in gyrA and no mutation in the parC gene. In all the phenotypic and genotypic properties of the 18 S. Enteritidis and 8 S. Typhimurium based on PFGE, phage types and antibiotic resistance pattern, the predominant patterns were XEI/BEI-PT32a-NaR (n = 5) and XTI/BTI-RNDC-no resistant antibiotics (n = 6), respectively.

The aim of this study was to give some insights into the prevalence, serovars, phage types, and antibiotic resistances of Salmonella from animal origin in the United Arab Emirates. Data on diagnostic samples from animals (n = 20,871) examined for Salmonella between 1996 and 2009 were extracted from the databases of the Central Veterinary Research Laboratory in Dubai and from typed strains (n = 1052) from the Robert Koch Institute, Wernigerode Branch in Germany and analyzed for general and animal-specific trends. Salmonella was isolated from 1,928 (9 %) of the 20,871 samples examined. Among the 1,052 typed strains, most were from camels (n = 232), falcons (n = 166), bustards (n = 101), antelopes (n = 66), and horses (n = 63). The predominant serovars were Salmonella Typhimurium (25 %), Salmonella Kentucky (8 %), followed by Salmonella Frintrop (7 %), and Salmonella Hindmarsh (5 %). When analyzed by animal species, the most frequent serovars in camels were Salmonella Frintrop (28 %) and Salmonella Hindmarsh (21 %), in falcons Salmonella Typhimurium (32 %), in bustards Salmonella Kentucky (19 %), in antelopes Salmonella Typhimurium (9 %), and in horses Salmonella Typhimurium (17 %) and S. Kentucky (16 %). Resistance of all typed Salmonella strains (n = 1052) was most often seen to tetracycline (23 %), streptomycin (22 %), nalidixic acid (18 %), and ampicillin (15 %). These data show trends in the epidemiology of Salmonella in different animal species which can be used as a base for future prevention, control, and therapy strategies.

We report on a coinfection of Mycobacterium avium subsp. avium and Salmonella enterica serotype Typhimurium var. Copenhagen phage type DT2 in pigeons from one flock, from which squabs were occasionally consumed by humans. Triplex quantitative real-time PCR and culture methods were used for M. a. avium detection in livers and culture method was used for the detection of Salmonella sp. in samples of liver and caecum of 33 examined birds. M. a. avium was detected in a total of 31 (93.9%) and Salmonella Typhimurium in a total of 11 (33.3%) pigeons. Coinfection with both pathogens was found in 10 (30.3%), infection with Salmonella Typhimurium alone in 1 (3.0%), and infection with M. a. avium alone in 21 (63.7%) pigeons. Neither pathogen was detected in one pigeon. There was no difference in clinical symptoms exhibited by pigeons infected by M. a. avium and/or Salmonella Typhimurium. All Salmonella Typhimurium isolates were sensitive to all 15 antimicrobials tested. According to these results we emphasize good heat treatment of consumed squabs.

Background: There is interest in applying bacteriophages to control Salmonella in pig production and pork processing. The following reports on the prevalence of Salmonella infecting bacteriophages within Ontario pig farms and associated with the holding area of a pork slaughterhouse., Results: Salmonella infecting bacteriophages were present in 30 and 28 of the effluent manure samples collected from 36 farms using S. Typhimurium DT104 or S. Heidelberg as host cell respectively. Bacteriophages were recovered in 95-100% of the 48 samples taken from holding pens within a high capacity slaughterhouse over a 12 month period. Bacteriophages isolated from farms exhibited similar host ranges which differed to that of slaughterhouse isolates. Salmonella (n = 21) from the slaughterhouse were susceptible to the endogenous bacteriophages. Despite being susceptible to the resident phages, the Salmonella populations were found to be genetically stable with the same genotypes being recovered over successive visits. Salmonella isolated from the farms were frequently resistant to the endogenous phages., Conclusions: Bacteriophages are prevalent in the pig slaughterhouse environment although they do not have a significant impact on the genetic structure of Salmonella populations. However, there was evidence that the Salmonella population structure on farms is influenced by the presence of infecting phages., (Copyright © 2010 Society of Chemical Industry.)

Salmonellosis was diagnosed in garden birds from 198 incidents in Scotland between September 1995 and August 2008. Salmonellosis was essentially a disease of finches in the north of Scotland, but in the south of Scotland it was also a problem in house sparrows. Almost all of the incidents were caused by Salmonella Typhimurium phage types 40 or 56/variant, but regional variation in phage types was observed. In the north of Scotland, one phage type (DT 40) predominated, but in the south of Scotland two phage types were commonly isolated (DTs 40 and 56/variant, with the latter the more common of the two phage types). This regional difference was statistically significant for salmonellosis in greenfinches, chaffinches and 'other garden birds', but not for house sparrows. Different temporal patterns for different species of bird and different phage types were also observed within regions. These findings suggest that the epidemiology of salmonellosis in garden birds varies depending on the phage type of Salmonella and the species of garden bird, with additional regional differences depending on the wild bird populations and the phage types of Salmonella in circulation. An awareness of these differences will help when formulating guidelines aimed at reducing the impact of salmonellosis in garden birds.

Cattle Egrets have a worldwide distribution, feed in proximity to cattle and other domestic animals, and often nest in large colonies in urban woodlots. Over a 3-yr period, nestlings from five Cattle Egret colonies from Central Texas, USA, were surveyed for salmonellosis. Prevalence of infection ranged from 29% to 95%. Seventeen Salmonella enterica subsp. enterica serotypes were isolated, of which the 4,5,12:i-monophasic serotype predominated in cultures of both the digestive tract and pooled spleen and liver. Of 11 4,5,12:i-monophasic isolates phage typed, eight were determinate type 193. The 4,5,12:i-monophasic isolates were susceptible to all antibiotics tested and were highly invasive in the day-old chick infection model. Microscopic lesions were found in the livers of Cattle Egrets with systemic infections with the 4,5,12:i-monophasic serotype, suggesting that infections with this serotype may often be fatal. Twenty-nine serotypes were identified in 179 S. enterica subsp. enterica isolates from horses admitted to the Texas A&M University Veterinary Teaching Hospital in 2 yr following the Cattle Egret study. The 4,5,12:i-monophasic serotype was not isolated from horses, but 12 serotypes were isolated from both horses and Cattle Egrets. The temporal distribution of the horse cases suggested that Cattle Egrets and horses may be exposed to similar sources of Salmonella, but provided no evidence of transmission between these two species. Similar conclusions were drawn when Cattle Egret isolates were compared to isolates from feedlot and dairy cattle from Texas and surrounding states. Given that the Cattle Egret 4,5,12:i-monophasic serotype was highly invasive and other isolates of this serotype have been associated with food poisoning, it is likely that Cattle Egret colonies pose a health risk to humans living near them.

On a dairy farm the herdsman got a pustular dermatitis and in the herd in a year several abortions occurred, the somatic cell count of the bulk milk increased and the fertility results decreased. The cause appeared to be an infection in both the herdsman and the herd with Staphylococcus aureus type T543.

Salmonella enterica serovar Typhimurium (S. Typhimurium) isolated and identified from swine were subjected for the analysis of antibiotic resistance pattern and clinically important class 1 and 2 integrons. In addition, S. Typhimurium isolates exhibiting ampicillin, chloramphenicol, streptomycin, sulfamethoxazole, tetracycline and florfenicol (ACSSuTF) resistance pattern as described in most Salmonella enterica serotype Typhimurium definitive type 104 (DT104) were characterized by polymerase chain reaction. All the isolates were resistant to more than four antibiotics and showed the highest resistance to streptomycin (94.1%), followed by tetracycline (90.1%), ampicillin (64.7%), chloramphenicol (56.8%) and gentamicin (54.9%). MIC value for the ten isolates ranged between 0.125-2 mug/ml for ciprofloxacin. Among the beta-lactams used, only one of the isolate exhibited resistance to ceftiofur (MIC 8 microg/ml). Sixty eight percent of these multi drug resistance (MDR) S. Typhimurium isolates carried clinically important class 1 integron with 1kb (aadA) and/or 2kb (dhfrXII-orfF-aadA2) resistance gene cassettes. This study reports the increasing trend of multi drug resistance (MDR) S. Typhimurium with clinically important class 1 integron in pigs. In addition, emergence of the ACSSuTF-type resistance in S. Typhimurium PT other than DT104 may limit the use of resistance gene markers in its detection methods by PCR.

The objectives of the study were to investigate the phenotypic and genotypic characterization of the persistent Salmonella Enteritidis (S. Enteritidis) isolates in two integrated broiler chicken operations, with attention focused mainly on the epidemiological approach. In the distribution of virulence genes, Salmonella enterotoxin (stn), invading host cell (invA), and Salmonella plasmid virulence (spvC) genes were widely distributed among the S. Enteritidis irrespective of their source of isolation, and Salmonella fimbrial (sefC) and plasmid encoded fimbrial (pef) genes were present in 28 and 20 S. Enteritidis strains, respectively. A total of 5 different XbaI-PFGE types were obtained from 31 S. Enteritidis isolates. Twenty-one types were divided on the basis their PFGE pattern, phage type and antimicrobial resistance pattern determined. There was a significant difference in phenotypic and genotypic characterization by two integrated broiler operations. Also, 8 isolates shown susceptible to all antimicrobials and 11 isolates with resistance to nalidixic acid were partly classified by XbaI PFGE pattern and by the phage type.

Surveillance data for Salmonella enterica serovar Enteritidis incidents and isolations from food animals in Great Britain from 1990 to 2005 were analysed to detect any trends and provide the basis for a comparison between phage types (pt) and antimicrobial sensitivity patterns in human beings and animals. During 2001 to 2005 there was a decrease in incidents involving most species except ducks. Only the numbers of incidents involving pts 6, 6a, 9b and 14b (in ducks) and pts 6a and 13a (in mammals) increased significantly during this period, whereas there were 93 per cent fewer incidents involving pt 4 than in 1990 to 2000. After adjustment for pt, the isolates from ducks were more resistant to nalidixic acid, tetracyclines and sulfonamides, and were more likely to be multiresistant than isolates from chickens. Isolates from turkeys tended to be more resistant to sulfonamides than isolates from chickens. pts 1, 5a, 6, 6a and 35 had the highest level of resistance after adjusting for species. During 2001 to 2005 there was an increase in resistance among pts 1, 6 and 7, in most cases involving nalidixic acid.

The purpose of this study was to investigate the biological and genetic characterization of persistent Salmonella isolates in an integrated broiler chicken operation, in an attempt to elucidate the source of contamination. From the breeder farm, the hatchery, the broiler farm and the chicken slaughter house of an integrated broiler chicken operation, a total of 6 serotypes were observed. Although S. Heidelberg was not detected in the broiler farm, it was consistently found in the breeder farm, the hatchery and the chicken slaughter house. Also, S. Enteritidis and S. Senftenberg were found in the hatchery and the chicken slaughter house, and the hatchery and the broiler farm, respectively. S. Gallinarum and S. Blockley were found only in the broiler farm, and S. Virchow was only recovered in the chicken slaughter house. Isolated S. Heidelberg, S. Enteritidis and S. Senftenberg strains were divided into 3, 5 and 7 types, respectively, on the basis of all properties. Especially, S. Senftenberg isolates, divided into four types by their antimicrobial resistance patterns, were all obviously the XbaI PFGE pattern. Also, four S. Enteritidis isolates resistant to nalidixic acid showed a difference in phage type and PFGE pattern. Such a different pattern was shown despite Salmonella isolates originating from an integrated broiler operation, suggesting that further epidemiological studies on many integrated chicken companies in Korea are needed.

US Department of Agriculture Food Safety and Inspection Service (FSIS) data on Salmonella enterica serotype Enteritidis in broiler chicken carcass rinses collected from 2000 through 2005 showed the annual number of isolates increased >4-fold and the proportion of establishments with Salmonella Enteritidis-positive rinses increased nearly 3-fold (test for trend, p<0.0001). The number of states with Salmonella Enteritidis in broiler rinses increased from 14 to 24. The predominant phage types (PT) were PT 13 and PT 8, 2 strains that a recent Foodborne Diseases Active Surveillance Network (FoodNet) case-control study associated with eating chicken. FSIS is directing more sampling resources toward plants with marginal Salmonella control to reduce prevalence in products including broilers. The policy targets establishments with common Salmonella serotypes of human illness, including Salmonella Enteritidis. Voluntary interventions should be implemented by industry.

A case of fatal salmonellosis in a Eurasian eagle-owl (Bubo bubo) from Bursa Province (northwestern Turkey) is described. The organs of the bird were examined histopathologically and microbiologically. Macroscopic and microscopic findings were consistent with a Salmonella infection. Salmonella enterica subspecies enterica serovar Enteritidis (S. Enteritidis) phage type (PT) 21b was isolated from the liver and spleen in pure culture and from the intestine. The isolate was susceptible to amoxycillin/clavulanic acid, ampicillin, chloramphenicol, gentamicin, kanamycin, tetracycline, and trimethoprim/sulphamethoxazole. This is the first report of an isolation of salmonellae from a wild bird species from Turkey and the first time S. Enteritidis PT21b has been reported from Turkey.

Salmonella enterica subsp. enterica (S.) serovar Agona plays an important role in Brazil as causative agent of salmonellosis in food-producing animals - in particular, pigs and poultry - as well as in humans. A total of 45 S. Agona isolates collected from slaughter pigs at three different slaughterhouses in Southern Brazil was investigated in this study for their phenotypic and genotypic relatedness. For this, the antimicrobial susceptibility patterns and the phage types were determined. Molecular analysis included the determination of plasmid profiles as well as the analysis of XbaI- and BlnI-generated macro-restriction patterns. Moreover, a novel typing method called subtracted restriction fingerprinting (SRF) was successfully applied to the S. Agona isolates. Based on all properties determined, a dominant clonal group comprising 33 of the 45 isolates was identified. Members of this group were susceptible to all antimicrobials tested, did not carry plasmids, shared the same phage type and were closely related or even indistinguishable by their EcoRI-PauI SRF patterns as well as their XbaI and BlnI macro-restriction patterns. Members of this clonal group were identified at all 3 slaughterhouses at variable frequencies and originated from pig herds raised in 15 different cities in Southern Brazil which were located up to 450 km apart from each other. Since the S. Agona-carrying slaughter pigs were from various integrated production lines, the results of this study suggest that a specific clonal group of S. Agona had entered numerous pig production lines. This observation supports the requirement for the establishment of monitoring and control programmes in Brazil which should also include molecular techniques to better trace the dissemination of S. Agona and other Salmonella serovars in pigs and other food-producing animals.

A study of randomly selected dairy farms in England and Wales was made between October 1999 and February 2001 to estimate the prevalence and incidence of Salmonella serovars. The farms were enrolled through five milk-buying companies, which represented 63 per cent of the dairy farms in England and Wales, and they were sampled on up to four occasions (449 farms at visit 1, 272 farms at visit 2, 251 farms at visit 3 and 243 farms at visit 4). In total, 19,296 samples of pooled faecal pats and slurry were collected. The farm-specific prevalence of all serovars of Salmonella ranged from 12.1 per cent (95 per cent confidence interval [CI] 8.2 to 16.0 per cent) to 24.7 per cent (95 per cent CI 19.4 to 30.1 per cent) at each visit. The most common serovars identified were Salmonella Dublin (3.7 to 6.6 per cent farm-specific prevalence at each visit), Salmonella Agama (1.8 to 7.6 per cent) and Salmonella Typhimurium (2.6 to 4.1 per cent) The prevalence varied by region and month of sampling and increased in late summer. The incidence rate of all serovars of Salmonella was 0.43 (95 per cent CI 0.34 to 0.54) cases per farm-year at risk. There was no significant difference between the incidence rates of the common serovars S Typhimurium (0.07), S Dublin (0.06) and S Agama (0.13). A total of 29 Salmonella serovars were isolated. Few of the isolates were resistant to the 16 antimicrobial agents tested, except the isolates of S Typhimurium dt104, of which 67.9 per cent were resistant to at least five of them.

Biofilm production by 221 Staphylococcus aureus isolates from 45 dairy herds was evaluated. Isolates were from composite milk of 117 cows, from teat skin of 70 cows, and from 34 milking machine unit liners. Of S. aureus from milk samples, 41.4% were biofilm producers, as compared to 24.7 and 14.7% of the isolates collected from skin and liners. Pulsed field gel electrophoresis (PFGE) best categorized S. aureus biofilm producers as compared to phage typing and binary typing. PFGE types that were significantly associated with isolation from milk as opposed to teat skin or liners, had isolates that were more likely to produce biofilm than PFGE types that were isolated from milk, skin and liners at similar frequencies. By contrast, PFGE type A was significantly associated with isolation from teat skin and had few biofilm producers. PFGE type Q, which is exclusively a milk, isolate produced more biofilm as evidenced by absorbance values. Given S. aureus that are associated with milk are more likely to produce biofilm as compared to extramammary sources (teat skin and milking unit liners), suggests that biofilm production is a risk factor for infection.

An outbreak of salmonellosis in a population of hospitalized horses resulted in the closure of a teaching hospital for a period of 10 weeks. Fecal samples were collected from suspected cases and cultured for Salmonella. Salmonella isolates were characterized using antimicrobial susceptibility testing, pulsed-field gel electrophoresis (PFGE) and phage typing. Thirty-three cases of infection by a multidrug-resistant strain of S. typhimurium were detected. The index case was admitted on 26 August 1999. Fifteen (45%) cases occurred between April and June 2000. PFGE results suggested that this strain of S. typhimurium might have been introduced into the hospital environment by a foal presenting with diarrhea. The hospital was closed on June 13, and intensive environmental cleaning and disinfection were completed. Enforcement of infectious disease control protocols in hospitals and environmental and patient surveillance is needed to prevent outbreaks of salmonellosis.

A total of 444 samples of raw chicken meat (thighs, breasts, wings, livers, gizzards, hearts and ovaries) that retailed at 145 different supermarkets in 47 prefectures in Japan were examined for contamination with Staphylococcus aureus in association with its enterotoxigenicity. S. aureus was isolated from 292 (65.8%) of the samples, and from 131 of the 145 supermarkets. There was no significant difference in the detection rate of S. aureus according to the type of meat examined. About 80% of 714 isolates belonged to the poultry (57.1%) and human biotypes (22.1%). Seventy-eight (21.7%) of 360 isolates were enterotoxigenic and isolated from 78 samples in 53 supermarkets in 31 prefectures. Staphylococcal enterotoxins (SEs) produced were SEB (50 isolates), SEA (14), SEC (8), SED (2), SEA+SEB (2), and SEA+SEC (2). Most of the enterotoxigenic isolates belonged to the human and poultry biotypes, coagulase type VII, VIII or IV, and were lysed by phages of group III. Identical SE types, biotypes, coagulase types and pulsed-field gel electrophoresis (PFGE) patterns were shown in isolates from different types of meat at the same supermarket and from samples taken from different supermarkets in the same prefectures or in isolates from samples obtained from several different prefectures. Among the 50 SEB-producing isolates, 27 yielded three similar PFGE patterns that differed by only a few fragments, suggesting that they were closely related genetically. The three patterns were found in isolates of samples that retailed at 17 supermarkets in 11 prefectures, indicating that they may be disseminated among raw chicken meat in Japan.

Staphylococcus intermedius isolates from diseased and healthy dogs were examined for production of extracellular enzymes and toxins, and phage patterns. There were no significant differences between the two groups of isolates in the production rates of DNase, protease, lipase, gelatinase, hyaluronidase, hemolysins, protein A, and TSST-1, or in phage patterns. But the production rate of enterotoxins in isolates from diseased dogs was significantly higher than that in isolates from healthy dogs. PFGE analysis was performed with isolates from different body sites in individual dogs. In 3 of 6 healthy dogs, identical PFGE patterns were seen in isolates from the nares, external auditory meatus or skin. The remaining 3 dogs yielded isolates of different patterns. In 4 of 6 diseased dogs, identical patterns were seen in isolates from lesions as well as from the other normal sites.

An increase in the prevalence of Salmonella enterica serotype Typhimurium DT104 has been reported worldwide. This study examined the prevalence of this microorganism in poultry environmental samples from commercial layer flocks and pullet environments as well as the sensitivity and specificity of a PCR-based method, and multiple antibiotic resistance profile of Salmonella serogroup B isolates in relation to the serotype and phagetype reference method for the identification of Salmonella Typhimurium DT104. A total of 435 Salmonella isolates were obtained from poultry house environmental samples tested during a 20-month period representing a prevalence of 5.5%. Of these, 313 (72%) isolates were identified as Salmonella serogroup B isolates. These isolates were tested by a PCR-based assay, and for resistance to five antibiotics: ampicillin, chloramphenicol, streptomycin, sulfonamides, and tetracycline (ACSSuT) for the rapid identification of Salmonella Typhimurium DT104. Upon comparing the antibiotic resistance and PCR results with serotype and phage type data, the sensitivity and specificity for the identification of Salmonella Typhimurium DT104 of both methods were found to be 100%, and 99.6%, respectively. Both methods can be completed within 24 h after obtaining an isolate, while serotyping and phagetyping required more than 5 days to complete., (Copyright 2004 Elsevier B.V.)

Grazing-fed cattle were previously demonstrated to be reservoir of non-O157 Shigatoxigenic Escherichia coli (STEC) serotypes in Argentina. The acid-resistance of some STEC strains makes it reasonable to assume the presence in feedlot of particular STEC serotypes. Fifty-nine animals were sampled every 2 weeks during 6 months by rectal swabs. Twenty-seven of 59 animals (45.8%) were shown to be Stx2(+); 3/59 (5.1%) carried Stx1(+) and 7/59 (11.9%) were Stx1(+) Stx2(+). Among 44 STEC isolates, 31 isolates were associated to 10 O serogroups (O2, O15, O25, O103, O145, O146, O157, O171, O174, O175) and 13 were considered non-typable (NT). Six H antigens (H2, H7, H8, H19, H21, H25) were distributed in 21 isolates whereas 23 were non-mobile (H-). Seventeen of 44 strains (38.6%) were eaeA(+) and 14 (31.8%) harbored the 60MDa plasmid. The megaplasmid (Mp) and eaeA gene were simultaneously found in a limited number of serotypes belonging to the enterohaemorrhagic E. coli (EHEC). E. coli O157:H7 strains, isolated from four (6.8%) animals, corresponded to the Stx2(+), eaeA(+), Mp(+) pattern. Three O157:H7 strains belonged to phage type 4 and the other strain was atypical. Many serotypes isolated from grain-fed cattle (O2:H25, O15:H21, O25:H19, O145:H-, O146:H-, O146:H21, O157:H7, O175:H8) also differed from those isolated by us previously from grazing animals. The serotypes O15:H21, O25:H19 and O175:H8 had not been identified at present as belonging to STEC. This work provides new data for the understanding of the ecology of STEC in grain-fed cattle and confirms that cattle are an important reservoir of STEC.

A total of 292 bovine Staphylococcus aureus isolates obtained from the 1950s (86 isolates), 1992 (107 isolates), and 2000 (99 isolates) were examined for antimicrobial susceptibility and phage typing. The same types of S. aureus (80, 52, 3A, 3A/3C, 42E, 77) were found among the isolates from all three time periods, representing 43.3% of the typeable isolates. This indicates that the Danish S. aureus population related to bovine mastitis has remained relatively unchanged over the last 50 years. The occurrence of antimicrobial resistance has remained low in Denmark in comparison to other countries in Europe.

During the decade to 1999, the incidence of human infections with the zoonotic pathogen verocytotoxin-producing Escherichia coli O157 (VTEC O157) increased in England and Wales. This paper describes the results of a survey of 75 farms to determine the prevalence of faecal excretion of VTEC O157 by cattle, its primary reservoir host, in England and Wales. Faecal samples were collected from 4663 cattle between June and December 1999. The prevalence of excretion by individual cattle was 4.2 per cent (95 per cent confidence interval [CI] 2.0 to 6.4) and 10.3 per cent (95 per cent CI 5.8 to 14.8) among animals in infected herds. The within-herd prevalence on positive farms ranged from 1.1 to 51.4 per cent. At least one positive animal was identified on 29 (38.7 per cent; 95 per cent CI 28.1 to 50.4) of the farms, including dairy, suckler and fattening herds. The prevalence of excretion was least in the calves under two months of age, peaked in the calves aged between two and six months and declined thereafter. The phage types identified most widely were 4, 34 and 2, which were each found on six of the 29 positive farms.

This study was conducted to investigate the diversity of phage types and associations between penicillin resistance and phage types among 815 Staphylococcus aureus isolates from bovine mastitis in nine European countries and USA. All isolates were examined for susceptibility to antimicrobial agents and characterised by phage typing. Penicillin resistance was found among strains from all countries with an average occurrence of 32.4% (2-71.4%). A total of 76% of isolates were identifiable by phage typing and 144 different phage types were observed. The most predominant types were phage type 29 (11% of the 815 isolates), phage type 52 (5%), and phage type 80 (5%). Phage type 95 and 29/52/52A/80 were both distributed within seven countries. In the countries with the highest occurrence of penicillin resistance a reduced diversity of phage types and phage groups was observed. Phage group III was significantly associated with penicillin resistance in contrast to phage group I (P=0.0023) and phage complex-80 (P=0.0066). This study confirms that a large number of phage types of S. aureus cause bovine mastitis, but that some types predominate. In addition, these findings could indicate that the use of penicillin in the bovine environment has selected for specific types of S. aureus in countries with a high frequency of resistance.

Problem Addressed: Shiga toxin-producing Escherichia coli (STEC), have emerged as food poisoning pathogens which can cause severe diseases in humans., Objective: The aim of this study was to determinate the serotypes and virulence genes of STEC strains isolated from sheep in Spain, with the purpose of determining whether sheep represent a potential source of STEC pathogenic for humans., Methods and Approach: Faecal swabs obtained from 697 healthy lambs on 35 flocks in Spain during the years 2000 and 2001 were examined for STEC using phenotypic (Vero cells) and genotypic (PCR) methods., Results: STEC O157:H7 strains were isolated from seven (1%) animals in six flocks, whereas non-O157 STEC strains were isolated from 246 (35%) lambs in 33 flocks. A total of 253 ovine STEC strains were identified in this study. PCR showed that 110 (43%) strains carried stx(1) genes, 10 (4%) possessed stx(2) genes and 133 (53%) both stx(1) and stx(2). Enterohaemolysin (ehxA) and intimin (eae) virulence genes were detected in 120 (47%) and in 9 (4%) of the STEC strains. STEC strains belonged to 22 O serogroups and 44 O:H serotypes. However, 70% were of one of these six serogroups (O6, O91, O117, O128, O146, O166) and 71% belonged to only nine serotypes (O6:H10, O76:H19, O91:H-, O117:H-, O128:H-, O128:H2, O146:H21, O157:H7, O166:H28). A total of 10 new O:H serotypes not previously reported in STEC strains were found in this study. Seven strains of serotype O157:H7 possessed intimin type gamma1, and two strains of serotype O156:H- had the new intimin zeta. STEC O157:H7 strains were phage types 54 (four strains), 34 (two strains) and 14 (one strain)., Conclusions: This study confirms that healthy sheep are a major reservoir of STEC pathogenic for humans. However, because the eae gene is present only in a very small proportion of ovine non-O157 STEC, most ovine strains may be less pathogenic.

Four hundred and twenty-two calves were examined for intestinal carriage of Shiga toxin-producing Escherichia coli O157:H7 using conventional plating. Two (0.5%) E. coli O157 were recovered. They were compared with 96 Argentine strains of different origin by pulsed-field gel electrophoresis, phage typing and PCR-RFLP of stx2 genes. One strain isolated from a calf, was closely related with 18 strains of clinical origin.

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

In the autumn of 1995 the first outbreaks of enterohemorrhagic Escherichia coli O157:H7 including ca 100 human cases were reported in Sweden. From outbreaks in other countries it is known that cattle may carry these bacteria and in many cases is the source of infection. Therefore, the present study was performed to survey the Swedish bovine population for the presence of verotoxin-producing E. coli (VTEC) of serotype O157:H7. Individual faecal samples were collected at the 16 main Swedish abattoirs from April 1996 to August 1997. Of 3071 faecal samples, VTEC O157 were found in 37 samples indicating a prevalence of 1.2% (CI95% 0.8-1.6). All 37 isolates carried genes encoding for verotoxin (VTI and/or VT2), intimin, EHEC-haemolysin and flagellin H7 as determined by PCR. Another 3 strains were of serotype O157:H7 but did not produce verotoxins. The 37 VTEC O157:H7 strains were further characterised by phage typing and pulsed-field gel electrophoresis. The results clearly show that VTEC O157:H7 is established in the Swedish bovine population and indicate that the prevalence of cattle carrying VTEC O157:H7 is correlated to the overall geographical distribution of cattle in Sweden. Results of this study have formed the basis for specific measures recommended to Swedish cattle farmers, and furthermore, a permanent monitoring programme was launched for VTEC O157:H7 in Swedish cattle at slaughter.

The National Salmonella Reference Laboratory (NRL-Salm) of the Federal Institute for Health Protection of Consumers and Veterinary Medicine receives putative Salmonella isolates originating from animals, food, feed and the environment for typing. This report summarises the results of the sero- and phagetyping studies. In livestock S. Typhimurium DT 104 with 24% and S. Enteritidis PT 4 with 9% of all isolations predominate like in the human isolates. These sero- and phagetypes are frequently isolated from food as well and consequently can reach the consumer via the food chain.

We studied 232 Salmonella strains from horses with salmonellosis in The Netherlands, isolated in the period from 1993 to 2000 in order to provide insight in the dynamics of sero-, phage types (pt) and antibiotic susceptibilities over time. The strains were tested for susceptibility to seven antimicrobial agents using the agar diffusion method. In addition, the isolates were sero typed and Salmonella enterica subspecies enterica Typhimurium and Enteritidis strains were further phage typed. S. Typhimurium strains of phage type 506 and 401 (both classified as DT 104 in the English phage typing system) were additionally tested for their susceptibility to chloramphenicol (C), streptomycin (S) and sulfonamides (Su). Resistance was common against tetracycline and ampicillin. Most strains were susceptible to enrofloxacin (Enr) and ceftiofur (Cef). Resistance to tetracycline (T), kanamycin (K), ampicillin (A) and trimethoprim/sulfonamide (Sxt) combinations decreased from 1993 to 2000, whereas the resistance to gentamicin (G), ceftiofur and enrofloxacin was stable over time. S. Typhimurium was the predominant serovar and showed more (multiple) resistance compared to other Salmonella serovars. Sixteen different resistance patterns were found, with resistance to T alone and the combination of ACSSuT and AKSxtT being the most common. The multiresistant S. typhimurium phage type 506 (DT 104) was the most common phage type isolated from horses and most of these strains showed the pentadrug resistance pattern ACSSuT. The S. Typhimurium phage type 401 (DT 104) was also found frequently with an ASSuT resistance pattern. The most common S. Typhimurium phage types in horses corresponded with those found in humans, pigs and cattle in the same period in The Netherlands.

The prevalence of Salmonella serovars and their antimicrobial resistance patterns were investigated among Danish turkeys between 1995 and 2000, by sampling the flocks approximately 14 days before they were slaughtered. Within the flocks, the prevalence of salmonella varied from 7.1 per cent to 25 per cent, and 24 different serovars were detected. The five most prevalent, which accounted for 58.5 per cent of the isolates were Salmonella Heidelberg (16.2 per cent of the isolates), Salmonella Agona (15.8 per cent), Salmonella Derby (12.4 per cent), Salmonella Muenster (7.3 per cent) and Salmonella Anatum (6.8 per cent). In addition, a few rough isolates and isolates belonging to the antigenically incomplete formulae 6,7:-:- and 4,12:b:- were found. The level of antimicrobial resistance was low; the highest resistance was recorded to ampicillin (13.7 per cent) and streptomycin (9.0 per cent) followed by tetracycline (8.5 per cent), sulphonamides (7.7 per cent) and spectinomycin (4.7 per cent). Resistance to quinolones was very low: four isolates were resistant to nalidixic acid, and only one was resistant to enrofloxacin. No resistance was recorded to colistin, apramycin, ceftiofur, florfenicol, or amoxycillin with clavulanic acid. Only 24 isolates were resistant to two or more compounds in various combinations of up to six compounds; one Salmonella Havana isolate was resistant to six compounds. Six isolates were serovar Typhimurium, but none of them belonged to phage type DT104.

Staphylococcus intermedius from pigeons, dogs, foxes, mink, and horses, was characterized by pulsed-field gel electrophoresis (PFGE) to evaluate the use of this typing method for discriminating among strains. SmaI cut the chromosomal DNA into 7-13 fragments ranging from approximately 48 kb to 655 kb, with most of the detectable fragments being smaller than 172 kb. S. intermedius from various animals had a high degree of restriction fragment length polymorphism. Pigeon strains have a similar genotype, despite the difference in their isolation area. Phage typing indicated that the dog, fox, and mink strains belong to the canine I or canine II type. The PFGE method further differentiated the mink strains from the dog and fox strains with regard to three fragments between 256 kb and 570 kb. As such, genomic DNA fingerprinting by PFGE appears to be an effective technique for discriminating S. intermedius strains from various animals. A combination of PFGE typing and phage typing would provide more detailed information than the single method for ecological investigations of S. intermedius.

Five hundred sixty-nine Salmonella were isolated out of 4745 samples from poultry products, poultry, and poultry environment in 1999 and 2000 from the Pacific northwest. These Salmonella were identified to their exact source, and some were serogrouped, serotyped, phage typed, and tested for antibiotic sensitivity. Food product samples tested included rinse water of spent hens and broilers and chicken ground meat. Poultry environment samples were hatchery fluff from the hatcheries where eggs of grandparent broiler breeders or parent broiler breeder eggs were hatched and drag swabs from poultry houses. Diagnostic samples were of liver or yolk sac contents collected at necropsy from the young chicks received in the laboratory. Of these samples tested, 569 were Salmonella positive (11.99%). Ninety-two Salmonella were serogrouped with polyvalent somatic antisera A-I and the polymerase chain reaction. Somatic serogroups B and C comprised 95.25% of all the Salmonella. Out of a total of 569 positive samples, 97 isolates of Salmonella were serotyped. A total of 16 serotypes and an unnamed Salmonella belonging to serogroup C1 were identified. The Salmonella serotypes were heidelberg (25.77%); kentucky (21.64%); montevideo (11.34%); hadar and enteritidis (5.15% each); infantis, typhimurium, ohio, and thompson (4.12% each); mbandaka and cerro (3.09% each); senftenberg (2.06%); berta, istanbul, indiana, and saintpaul (1.03% each); and an unnamed monomorphic Salmonella (2.06%). Ninety-two Salmonella were tested for drug sensitivity with nine different antimicrobials. All of the 92 Salmonella were resistant to erythromycin, lincomycin, and penicillin except one sample (S. berta), which was moderately sensitive to penicillin. All of the tested Salmonella were susceptible to sarafloxacin and ceftiofur. The percentages of Salmonella susceptible to sulfamethoxazole-trimethoprim, gentamicin, triple sulfa, and tetracycline were 97.83%, 92.39%, 86.96%, and 82.61%, respectively.

The Paul Ehrlich Institute, Langen, in Germany has been licensed different live vaccines of S. Typhimurium and S. Enteritidis for use in the veterinary medicine since I the 90s. The Robert Koch Institute has established a lab-based surveillance system for these live vaccine strains for an evaluation of recent public health safety. Since 2000 all strains of S. Typhimurium and S. Enteritidis from humans were investigated in respect to their phage types and other vaccine markers. 3676 S. Typhimurium strains and 4489 S. Enteritidis strains mainly from Salmonellose patients were investigated after phage typing according to their auxotrophic or antibiotic resistance markers. The live vaccine strains of Zoosaloral, TAD Salmonella vacT or TAD Salmonella vacE and Salmovac SE could not be found from infections in humans.

One hundred and twenty 1-day-old broiler chickens were divided into four groups: group I unmedicated and orally challenged with 1.5 x 10(8) cfu of Salmonella enteritidis phage type 4; group F infected and treated with 300 ppm of fosfomycin in their drinking water; group CF uninfected and treated, and group C maintained as a control group. Their performance, clinical signs, S. enteritidis PT4 reisolation and biochemistry variables were compared. Group F showed fewer symptoms and gross lesions than those from group 1. Fosfomycin treatment at 300 ppm improved body weight at 7 days of age by 42.3%. S. enteritidis PT4 reisolation in group I was higher than in the treated group, but total decontamination of challenged birds was not achieved. There was an increase in the levels of total protein and globulins in group I but not in the treated group. Fosfomycin caused no adverse effects on chickens from group CF, assessed by performance and biochemical variables. The results indicate that fosfomycin could be used in the treatment of S. enteritidis PT4 experimental infection.

Salmonella enteritidis (SE) is an important cause of egg-associated outbreaks in both Europe and the United States. Phage typing has become an important epidemiologic tool in identifying the source of outbreaks. Limitations of phage typing have become apparent with wholesale egg distributors that have multiple suppliers in an area where a particular phage type is endemic. Several different molecular typing methods were evaluated for their discriminatory power to identify genetic differences among different SE phage types isolated in Europe and the United States. Pulsed-field gel electrophoresis (PFGE) identified a single DNA pattern among the different SE phage types. Comparison of the nucleotide sequence for several Salmonella virulence genes failed to identify a single nucleotide change in the gene sequences from most SE isolates, regardless of phage type. On the basis of these results, the different SE phage types appear to be genetically related or clonal. However, with primers 1283 and Opa4, it was possible to differentiate not only SE isolates from different geographic locations but those within a specific geographic locale as well by random amplified polymorphic DNA polymerase chain reaction. Any chance for discerning genetic differences among isolates will need to rely on molecular techniques other than PFGE.

A study on the prevalence of the faecal carriage of Verocytotoxin (VT)-producing Escherichia coli (VTEC) O157 and on the rate of carcass contamination was carried out on feedlot cattle and dairy cows at slaughter in northern Italy. Between April 1998 and January 1999, 12 sampling visits were performed on different days in seven different slaughterhouses. At each visit, 5-12 animals consecutively slaughtered were selected. From each animal, faeces were collected from the rectum immediately after slaughter and surface swabs were taken from the leg region and the diaphragmatic insertion of the carcass. All samples were examined for the presence of VTEC O157 using an immunomagnetic separation technique. A total of 100 animals coming from 60 different farms were examined. In total, VTEC O157 was isolated from the intestinal content of 17, and from the carcasses of 12 of the 100 animals examined. In particular, VTEC O157 was recovered from six (35.3%) out of the 17 carcasses from which the organism had previously been isolated from rectal content and from six (7.3%) of the 82 carcasses of the stool-negative cattle. In seven carcasses, VTEC O157 was isolated from the leg area, in two carcasses from the diaphragmatic area, and in three carcasses from both areas. Major differences in the prevalence of VTEC O157 were observed in the different groups of cattle sampled. In 7 of the 12 sampling visits, all the specimens examined were negative, while 16 of the 17 positive stool samples and 11 of the 12 positive carcass swabs were collected during three of the visits, performed in June in three different abattoirs. In these three visits, the ratios between the percentage of animals carrying VTEC O157 in the stools and the percentage of contaminated carcasses were 0.33, 0.57, and 1.66, respectively; thus, confirming that slaughter practices can largely influence the rate of carcass contamination. Phage typing and PFGE analysis of VTEC O157 isolated from samples collected at the same visit suggested that both auto- and cross-contamination occurred.

Escherichia coli O157 isolates from cattle in Japan (n = 91) and in the USA (n = 415) were compared by pulsed-field gel electrophoresis of endonuclease-cleaved genomic DNA, location of the stx genes and bacteriophage typing. Three isolates from cattle in Japan with high similarity to isolates from cattle in the USA were found. Isolates from cattle farms in Japan and the USA may share a common source.

The clonal relationship among Salmonella enterica serotype Typhimurium isolates from selected pig production units in Denmark was investigated by the pulsed field gel electrophoresis (PFGE) typing method to determine environmental survival and spread of Salmonella in different herds. Thirty-four Typhimurium isolated during 1996-1998 from porcine faeces and environmental samples from three pig farms designated 1, 3 and 5 were characterised by PFGE using two restriction enzymes. Farm 5 supplied piglets to farm 1 and the herds were located close to each other. Results of PFGE analysis showed both intra- and inter-relationships, i.e. identical PFGE patterns among the faecal and environmental isolates from farm 1 and farm 5. All the isolates from farm 3 irrespective of the source showed identical PFGE patterns, but were different from samples from farms 1 and 5. This study indicates spread between farms and survival of a farm-specific clone. Furthermore, identical PFGE patterns of isolates from piglet supplier and finisher herds indicate that the farrow-to-grower herd of farm 5 was sub-clinically infected prior to delivery to farm 1 and thereby caused the transmission of Salmonella.

The prevalence of resistance to florfenicol, a phenicol drug newly introduced in veterinary therapy, was determined in 86 chloramphenicol-resistant Salmonella Typhimurium isolates from cattle collected during 1985-1995. All were highly resistant to chloramphenicol (MICs > or = 128 mg/L) and 38 were simultaneously resistant to florfenicol (MICs >16 mg/L) and to beta-lactam agents, spectinomycin, streptomycin, sulphonamides and tetracyclines. The isolates susceptible to florfenicol harboured the chloramphenicol acetyl transferase gene, cat of type I. All the florfenicol-resistant isolates harboured the floR resistance gene and the characteristic multiple resistance genetic locus, previously characterised in a S. Typhimurium DT104 strain and identified by a multiplex PCR. Plasmid profiles and ribotype patterns were determined for all the isolates. The florfenicol-resistant isolates were grouped into the same ribotyping pattern and presented similar plasmid profiles, whereas the florfenicol-susceptible isolates showed a wider genetic diversity that is usual for S. Typhimurium. Thus, the florfenicol-resistant isolates could represent a clonal cluster, closely related to, if not of DT104 phage type, which appeared in 1989 and is now predominant within chloramphenicol-resistant S. Typhimurium. The multiplex PCR provided a useful tool to survey further evolution of multiresistant S. Typhimurium strains.

Different subtypes of Listeria monocytogenes were isolated from various animal and environmental samples during an episode of increased mortality on a fallow deer (Dama dama) farm. During a 4-wk period, six fallow deer died, including four does, one fawn, and one adult buck. Prior to death, one of the does had exhibited central nervous system signs characteristic of listeriosis. Postmortem examination of the six deer showed no histologic changes typical of listeriosis, although inflammatory changes were present in several organs. Different subtypes of L. monocytogenes were isolated from brain samples from six deer, from fodder and soil from the deer feeding area, and from faces of some healthy animals on the farm. Listeria monocytogenes, which was frequently isolated in the environment of the farm, was considered the probable major cause of mortality in these fallow deer.

The authors describe the source and prevalence of pathogenic Salmonella serovars among poultry farms in Saudi Arabia. A total of 1,052 (4%) Salmonella isolates were recovered from 25,759 samples of poultry (broilers, layers, broiler breeders and layer breeders) and poultry environments (box liner, litter, drag swab, droppings, mice and feed) were examined bacteriologically between 1988 and 1997 at the Poultry Disease Laboratory at the National Agriculture and Water Research Center in Riyadh. Eleven Salmonella serogroups representing 38 different Salmonella serovars were identified by means of antigenic analysis. The majority of the 276 isolates (26.2%) of Salmonella typed, were recovered from liver, heart and intestines of the broilers and layers. The most prominent Salmonella serogroups isolated were as follows: serogroup C1 (392 isolates, 37.26%), B (289 isolates, 27.47%) and D1 (269 isolates, 25.69%). However, untypable and multiple serogroups were also encountered, the most frequent isolates serotyped belonged to groups C1 (97 isolates, 24.7%), D1 (86 isolates, 31.9%), and B (71 isolates, 24.6%). Salmonella Enteritidis (85 isolates, 98.8%), Salmonella Virchow (48 isolates, 57.8%), Salmonella Paratyphi B var. Java (41 isolates, 57.7%) and Salmonella Infantis (30 isolates, 20.6%) were distributed the most widely as all were encountered in poultry and in poultry environments. S. Enteritidis phage type 4 (30 isolates, 35.3%), was the phage type most frequently detected among group D1 phage types, while 39 (45.8%) of the isolates of S. Enteritidis could not be phage typed.

A 6-month longitudinal study was conducted on 30 dairy cows in early lactation and their human handlers on six farms across Trinidad. Weekly samples of bulk milk, composite milk and anterior nares and hand swabs from human handlers were collected and cultured for Staphylococcus aureus on Baird-Parker agar (BPA). The susceptibility of S. aureus strains to bacteriophages and the relatedness of strains isolated over the study period were determined. Sixty-three (51.2%) of 123 strains of S. aureus from bulk milk were typable compared with 111 (57.3%) of 194 and 82 (61.7%) of 133 strains isolated from composite milk and human handlers, respectively. The differences were not statistically significant (P > 0.05; chi 2). Bovine phage 42D lysed 3.3% (4 of 123), 16.5% (32 of 194) and 12.0% (16 of 133) of S. aureus strains isolated from bulk milk, composite milk and human handlers, respectively. The differences were statistically significant (P < 0.001; chi 2). Amongst bulk milk isolates of S. aureus, 35 (31.8%) of 110 exhibited relatedness in 11 groups based on their phage patterns and groups. The mean maximum interval between the first and last detection of related S. aureus strains in a group was 11.5 +/- 7.3 weeks. Amongst composite milk strains of S. aureus, 23 (46.0%) of 50, 25 (62.5%) of 40 and 22 (53.7%) of 41 exhibited relatedness on farms IB 2, IB 27 and IC 23, respectively, but the differences were not statistically significant (P > 0.05; chi 2). On farm IB 2, five groups of related strains of S. aureus were detected with a mean maximum interval of detection of 18.2 +/- 8.5 weeks compared to farm IB 27 where five groups of related strains were also observed but with an interval of 13.8 +/- 8.2 weeks. On farm IC 23, a total of seven groups of related S. aureus strains were detected with a mean interval of 8.0 +/- 5.5 weeks. For human strains of S. aureus from farm IB 2, nine (56.3%) of 16 strains isolated from anterior nares exhibited relatedness in three groups with a mean maximum interval of 13.3 +/- 4.7 weeks compared to four (25.0%) of 16 hand swab isolates which exhibited relatedness in two groups with mean interval of detection of 11.0 +/- 1.4 weeks. The differences were not statistically significant (P > 0.05; chi 2). On farm IB 27, for anterior nares isolates, eight (72.7%) of 11 exhibited relatedness in two groups with a mean maximum interval of detection of 20.5 +/- 2.1 weeks compared to hand swab isolates, with six (50.0%) of 12 showing relatedness in two groups and a mean interval of 10.5 +/- 2.1 weeks. It was concluded that dairy cows and their human handlers carried particular strains of S. aureus at various sites for extended periods, which served as continuous sources of contamination of milk and may play a significant role in the occurrence of subclinical mastitis, with an obvious economic impact.

The recent isolation of phage type 4 Salmonella enteritidis strains from poultry and humans in the United States has generated considerable concern because this phage type is predominant in both animals and humans in many other nations. Understanding whether the presence of these strains in poultry flocks poses an elevated threat to public health is a critical issue for developing effective disease control programs. The present study evaluated whether S. enteritidis strains of various phage types found in poultry in the United States (phage types 4, 8, and 13a) differed in their potential for horizontal transmission from experimentally infected chicks to uninoculated chicks housed in the same isolator units. Five days after two seeder chicks in each group of 12 were inoculated with oral doses of approximately 10(3) S. enteritidis cells at 8 days of age, ceca and livers were sampled from seeder chicks and from their contact-exposed penmates. On the basis of the detection of S. enteritidis in cecal samples, phage type 4 strains were transmitted horizontally at a significantly lower frequency than were strains of other phage types. Nevertheless, two of three phage type 4 strains evaluated were very highly invasive.