Herd-level diagnosis for Salmonella enterica subsp. enterica serovar Dublin infection in bovine dairy herds

Herd-level sensitivities of bacteriological and serological methods were compared in 79 bovine dairy herds, recently infected with Salmonella enterica subsp. enterica serovar Dublin. All farms experienced clinical signs of salmonellosis for the first time and had no history of vaccination against salmonellosis. At the start of the study, infection with serovar Dublin was confirmed with at least one positive bacteriologic culture for serovar Dublin from a clinical case (gold standard for herd infection).Bacteriological culture was done on samples of dung-pits, drinking water, bulk-milk filters, and faeces of animals with current or earlier clinical signs of salmonellosis. Blood samples of all animals and bulk-milk samples were tested using an ELISA.Herd-level sensitivity (HSe) of culture of dung-pits, drinking water, bulk-milk filters, and faeces of animals with current or earlier signs of salmonellosis was 45, 5, 7, and 38%, respectively. HSe for serology of all animals was 100%. If blood samples of all calves 4-6 months old were examined, at least one calf was seropositive on 91% of the infected farms. If serology was performed on samples of animals with current or earlier signs of salmonellosis, at least one animal was seropositive on 80% of the infected farms. HSe for bulk-milk samples was 54%. However, if clinical signs of salmonellosis were observed only in lactating animals, sensitivity of bulk-milk serology was 79%.Interesting combinations of methods were the combination of serology of bulk milk with either serology of animals with current or earlier signs of salmonellosis (HSe=91%), or serology of all calves of 4-6 months old (HSe=99%).     

Salmonella enterica infections in Spanish swine fattening units

The present study is the first conducted in Spain to estimate the bacteriological herd prevalence of Salmonella enterica in fattening units and to describe the Salmonella serovar diversity on these farms using a sample representative of the entire swine population. For this purpose, 10 faecal samples were collected from 10 different pens containing pigs close to market weight in a total of 232 fattening units. Total sample size was proportionally distributed according to the fattener census in each of the regions of the country and all the samples were examined by culture of 25 g of faecal material. One hundred (43.1%) farms had at least one Salmonella-positive sample (95% CI: 37-49.1%). Salmonella enterica was detected in 290 (12.5%) pooled faecal floor samples (95% CI: 11.2-13.8%). The apparent herd prevalence of salmonellosis was similar among multi-site, finishing and farrow to finish farms. Overall, 24 different serovars were identified, with S. Typhimurium, S. Rissen and S. Derby being the most common both at herd and sample level. Results of phage typing were available for the 91 isolates of S. Typhimurium. A total number of 10 different phage types were identified, with DT 193 being the most frequent. Phage types DT 104, DT 104b and DT U302, which have been associated with several multi-resistant patterns, accounted for 23% and 29% of the Typhimurium total isolates or Typhimurium infected farms respectively.     

Risk factors for clinical salmonellosis in Virginia, USA cattle herds

Veterinarians working in several Virginia, USA counties noticed an increase in clinical salmonellosis in cattle herds during 1994. We conducted a case–control study to identify potential risk factors for salmonellosis in cattle herds in this region. The potential impact of exposure to poultry-related factors was of particular interest because of the close proximity of poultry and cattle operations in this part of the state. Information was collected from 23 case farms and 23 control farms matched by herd veterinarian, type of enterprise (beef vs. dairy), and location. Faecal and environmental samples were collected during herd visits for bacteriologic culture.

Salmonella was isolated from 4.7% of 531 faecal, feed, water, and environmental samples collected. Salmonella Typhimurium was the serotype found most frequently in clinical cases in participating herds and from samples collected during study visits. Case herds had a higher percentage of study samples positive for Salmonella. Potential risk factors were screened using methods for pair-matched data and then analyzed in individual conditional logistic-regression models to estimate odds ratios with 95% profile likelihood confidence intervals. Significant factors included: the number of mature cows in the herd (OR=1.01; CI 1.00, 1.03), percent change in the number of mature cows during 1994 (OR=0.68; CI 0.11, 0.96), having calves usually born in a building rather than outdoors (OR=0.17; CI 0.01, 0.98), poultry manure spread on bordering property (OR=4.00; CI 1.00, 26.50), signs of rodents in cattle-housing or feed-storage areas (OR=2.75; CI 0.94, 9.92), and contact of wild geese with cattle or feed (OR=4.5; 1.16, 29.51).     

Sero types, phage types and antibiotic susceptibilities of Salmonella strains isolated from horses in The Netherlands from 1993 to 2000

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.     

Molecular characterization, spread and evolution of multidrug resistance in Salmonella enterica typhimurium DT104.

Multidrug-resistant Salmonella enterica serovar Typhimurium phage type DT104 has emerged during the last decade as a global health problem because of its involvement in diseases in animals and humans. Multidrug-resistant DT104 strains are mostly resistant to ampicillin, chloramphenicol, streptomycin, sulfonamides and tetracyclines (ACSSuT resistance type). The genes coding for such resistances are clustered on the chromosome. This paper reviews new developments in the characterization of S. enterica Typhimurium DT104, its chromosomal antibiotic resistance genes and their spread among other S. enterica Typhimurium phage types and other S. enterica serovars, the development of specific detection methods, virulence characteristics, and the evolution of multidrug-resistance with regard to the emergence of quinolone resistance.     

Effect of previous antimicrobial treatment on fecal shedding of Salmonella enterica subsp. enterica serogroup B in New York dairy herds with recent clinical salmonellosis

The association of herd- and sample-level factors with the isolation of Salmonella group B from cattle fecal samples was analyzed. Study farms were 65 dairy herds with a recent history of laboratory-confirmed clinical salmonella infections. Herds were visited once per month for three months to collect data and samples for bacteriological culture. Herd size varied widely from 34 to 3700 total cattle on the farm (median=370). Salmonella serogroup B was isolated from 270 of 2726 samples tested. The predominant serotypes identified were S. Typhimurium and S. Typhimurium var. Copenhagen. Logistic regression was used to analyze the relationship between potential risk factors and isolating Salmonella serogroup B. The only herd-level factor which was significantly associated with fecal shedding was total herd size (hundreds of cattle OR=1.09; 95% confidence interval (CI): 1.05, 1.14). The probability of a positive sample decreased substantially for longer intervals between the initial clinical case and sampling (interval in months OR=0.5; 95% CI: 0.3, 0.6). The presence of diarrhea increased the risk of shedding (OR=2.1; 95% CI: 1.4, 3.0). The effect of recent treatment with antimicrobial agents depended on age group. For heifers and cows, recent antimicrobial treatment increased the probability of isolating Salmonella (heifers OR=11.8; 95% CI: 2.9, 48.8; cows OR=4.1; 95% CI: 2.0, 8.4), but this effect was not statistically significant for calves before weaning. Among animals without recent antimicrobial treatment, preweaned calves were more likely to have positive samples than cows (OR=3.5; 95% CI: 1.8, 6.9; heifers OR=4.7; 95% CI: 2.3, 9.6).     

Phenotypic and genetic characterization of Salmonella enterica subsp. enterica serovar Typhimurium isolated from pigs in Rio Grande do Sul, Brazil

This study aimed to investigate the relatedness of porcine Salmonella enterica subsp. enterica (S.) serovar Typhimurium strains isolated in Southern Brazil. Sixty-six isolates from pigs belonging to three commercial companies were submitted to phage typing, XbaI-macrorestriction (PFGE), IS200 hybridization, rep-PCR, antimicrobial susceptibility testing, and PCR assay targeting the spvR region. All strains presented a unique rep-PCR pattern and 63 strains had a common IS200 profile. One pulse-type (XA) was the most prevalent (39/66 strains) and included strains of phage types DT177, DT192, DT194 and RDNC. The spvR region was detected in three strains, which harboured plasmids of 90 kb. High rates of tetracycline, sulfonamide and streptomycin resistance were found. Isolates from farms located in different geographic regions but associated to the same commercial companies clustered together and presented a common resistance profile. Results suggested that clonal groups of S. Typhimurium are present in pig commercial companies in Southern Brazil.     

Factors related to the incidence of clinical encephalomyocarditis virus (EMCV) infection on Belgian pig farms

We set up a matched case-control study of potential risk factors for clinical encephalomyocarditis virus (EMCV) in 58 pig farms in West Flanders (Belgium). In total, 29 farms experienced a clinical outbreak of EMCV confirmed by EMC virus isolation. Mortality was seen only among suckling piglets (18 case farms), in piglets and other age-groups (4 case farms), or only among fattening pigs (7 case farms). Five farms had reproductive problems among the sows. Control farms were matched geographically on farm size and farm type and were selected on the absence of clinical signs. A questionnaire on potential risk factors for EMCV was developed to collect data at both case and control farms. The exploration of the data used clusters of factors associated with clinical EMCV infection: (a) rodents, (b) general farm set up and (c) general hygiene. The multivariable relationships between clinical appearance of EMCV and potential risk factors were tested with conditional logistic regression. The final model on all farms contained presence of mice (OR=8.3) as a risk factor for clinical EMCV infection while the flow of manure up through the slatted floor (OR=0.11) and movement of manure between manure pits in the pig stable (OR=0.14) were protective.     

Oral administration of Salmonella enterica serovar Typhimurium expressing swine interleukin-18 induces Th1-biased protective immunity against inactivated vaccine of pseudorabies virus

Enhancing and/or modulating innate and adaptive immunity by cytokines appears to be greatly useful to provide effective protective immunity against infectious diseases. However, an effective delivery system for mass administration in livestock industry is needed because of limitations such as cost, labor, time, and protein stability. Here the immunomodulatory functions of swine interleukine-18 (swIL-18), known as IFN-γ-inducing factor (IGIF), were evaluated in a vaccination model of pseudorabies virus (PrV) using attenuated Salmonella enterica serovar Typhimurium as the oral delivery system. The oral administration of S. enterica serovar Typhimurium expressing swIL-18 prior to vaccination with inactivated PrV vaccine induced enhanced levels of serum PrV-specific IgG and its IgG2 isotype, compared to administration of S. enterica serovar Typhimurium harboring the empty vector. Furthermore, S. enterica serovar Typhimurium expressing swIL-18 mounted Th1-biased cellular immune responses against PrV antigen, as evaluated by the production of IFN-γ and IL-4 from peripheral blood mononuclear cells of piglets. Subsequently, Th1-biased immunity induced by S. enterica serovar Typhimurium expressing swIL-18 showed rapid response and rendered piglets displayed more alleviated clinical signs following the virulent PrV challenge. Also, this alleviation of clinical signs was further confirmed by the reduction of nasal excretion of PrV after challenge. The present study demonstrates the extended use of immunomodulatory functions of swIL-18 orally delivered by attenuated S. enterica serovar Typhimurium.     

The attenuated sopB mutant of Salmonella enterica serovar Typhimurium has the same tissue distribution and host chemokine response as the wild type in bovine Peyer's patches

Salmonella enterica serovar Typhimurium is an important cause of enteric infections in farm animals and it is one of the most frequent food borne infections worldwide. Serovar Typhimurium lacking the sopB gene is attenuated for induction of host inflammatory response and fluid accumulation into the intestinal lumen, which correlates with clinical diarrhea. SopB is an inositol phosphate phosphatase, but its exact role in the pathogenesis of salmonellosis is still unclear. We employed the bovine ileal ligated loop model to compare the tissue distribution of a sopB mutant and its wild type parent serovar Typhimurium. Sections of the Peyer's patches were histologically processed and immuno-stained for detection of serovar Typhimurium. In addition, samples were processed for transmission electron microscopy, and the profile of expression of host chemokine and cytokine responses was assessed. Ultrastructurally both strains had the same ability to invade intestinal epithelial cells. No differences were detected in the tissue distribution of the sopB mutant and the wild type organism and both strains elicited the same profile of chemokines and pro-inflammatory cytokines. In conclusion, our results indicate that the attenuation of the sopB mutant is associated with pathogenic mechanisms other than invasion and distribution in host intestinal tissues.     

Impact of Salmonella Typhimurium DT104 virulence factors invC and sseD on the onset, clinical course, colonization patterns and immune response of porcine salmonellosis

The present study was conducted to study the impact of the virulence factors invC and sseD of the two type III secretion systems of Salmonella enterica serovar Typhimurium (S. Typhimurium) on the pathogenesis of the porcine S. Typhimurium DT104 infection. For this purpose, two S. Typhimurium mutant strains with a disrupted invC gene of the Salmonella pathogenicity island 1 or with a disrupted sseD gene of the Salmonella pathogenicity island 2 have been studied in experimental infection of pigs. Twenty-two 7-week-old male hybrid pigs were either infected with one of the mutants or the wild-type S. Typhimurium DT104 strain. Each group was examined for clinical signs, Salmonella shedding rate and the specific antibody response. Survival and replication were evaluated by qualitative and quantitative determination of the colonization rate. The humoral and cellular immune responses were examined using isotype-specific ELISA and quantitative real-time PCR of IL-2, IL-4, IL-10, IL-12 and IFN-gamma. The results proved that both mutants had a lower virulence (with marked differences between both mutants) than the wild-type and that both virulence factors have importance in porcine salmonellosis. Only pigs infected with the wild-type S. Typhimurium DT104 exhibited typical clinical symptoms of salmonellosis like anorexia, vomiting, disturbed demeanour, fever and diarrhoea. Deletion of the invC gene resulted in a significantly reduced colonization rate. Interestingly, the mRNA expression of both type-1 and type-2 cytokines were significantly decreased in pigs infected with either the invC-mutant and the sseD-mutant strain.     

Distribution of Salmonella serovars and phage types on 80 Ontario swine farms in 2004

The objective of this study was to describe the distribution of Salmonella spp. on Ontario grower-finisher pig farms. Eighty swine farms were visited from January through July 2004. On each farm, fecal samples were collected from 5 pens, 2 rectal samples and 1 pooled sample from fresh manure on the floor per pen. Salmonella was isolated from 91 (11%) of the 800 rectal samples and 73 (18%) of the 397 pooled samples. Overall, Salmonella was recovered from 37 (46%) of the 80 farms. On each positive farm, Salmonella was cultured from 1 to 7 pigs or 1 to 5 pens. Of the 37 farms, 18, 13, 5, and 1 yielded 1, 2, 3, and 4 serovars, respectively. The most common serovars were S. Typhimurium var. Copenhagen, S. Infantis, S. Typhimurium, S. Derby, S. Agona, S. Havana, and S. enterica subsp. I:Rough-O. The 3 most frequent phage types were PT 104, PT 104a, and PT 104b. There was a statistically fair agreement between samples collected directly from pigs and pooled pen samples in determining the Salmonella status at the pen and farm level (kappa = 0.6, P < 0.0001). However, in 62 pens, Salmonella status, serovars, or phage types differed between the pig and pooled pen samples. The distribution of Salmonella on the swine farms in this study indicates that, in developing an intervention strategy, priority should be given to farms positive for S. Typhimurium var. Copenhagen. Also, the variation in Salmonella status between pig and pooled pen samples deserves consideration in a sampling strategy.     

Nationwide surveillance of salmonella in the faeces of pigs in Japan

The prevalence of faecal carriage of salmonella in 5393 pigs reared on 218 pig farms located in 31 of 47 prefectures in Japan over the period July 2003 to June 2005 was investigated. We isolated 172 strains belonging to 20 serovars and one untypable Salmonella enterica from 169 pig faecal samples (3.1%) collected from 48 farms (22.0%). The most prevalent type of S. enterica was untypable O4,12:d:- which lacks phase 2 flagellar antigen, representing 29.1% (50/172) of all isolates. Of 26 S. enterica serovar Typhimurium isolates, 16 strains appeared to be definitive phage type 104 (DT104) by polymerase chain reaction.     

Epidemiology and characterization of animal Salmonella enterica subspecies Enterica serotype typhimurium DT104 in Hungary

Reports on the internationally emerging significance of multiresistant zoonotic Salmonella in animals and man prompted studies to estimate the significance of multiresistant Salmonella enterica subspecies enterica serotype Typhimurium (S. Typhimurium) phage type DT104 of animal origin in Hungary. A collection of 231 strains (primarily of goose, turkey, poultry and porcine origin from the years 1997-1998) was tested for resistance against 7 selected antibiotics (ampicillin, chloramphenicol, enrofloxacin, nalidixic acid, streptomycin, tetracycline and sulphamethoxazole). Strains with resistance against 3 or more were defined as multiresistant. All strains were phage typed using Felix-Callow's S. Typhimurium phage typing system, and 91 of them (suspect DT104) were also typed according to Anderson's definitive typing (DT) system. In this study, 14% of animal strains from 1997-1998 was classified as DT104, for which turkey, pig and duck seemed to be the main carriers, and the multiresistant non-DT104 strains represented a further 6% of this collection. The prevalence of DT104 was highest among strains of turkey origin (50%), followed by strains of pig (29%), chicken (25%), duck (19%), and goose (3%) origin. The other DT104 related phage types (DT12 and U302) were only detected in the case of 4 strains (2 of porcine, and one each of turkey and of goose origin). The DT104 corresponded to the Felix-Callow types 2/3 or 2c/3 in each case, except in the case of 3 turkey strains where they corresponded to type 35/3. Nalidixic acid resistance was detected in all multiresistant turkey strains and in some of other animal origin but none of these strains were resistant to enrofloxacin. A retrospective analysis (based on the above relationship) indicated that S. Typhimurium strains corresponding to DT104 could be present and increase in the Hungarian farm animal population from about 2% to 20% between 1985 and 1990, in a manner similar to the emergence of human DT104, as reported elsewhere (Pászti et al., 2000). The 91 suspect DT104 strains were also tested for plasmid profile and for spvC gene indicating the presence of the large serotype specific plasmid (Ssp). No characteristic plasmid profile could be attributed to S. Typhimurium DT104. The serovar-specific large plasmid was detected by PCR for spvC in 100% of DT104 strains and in 77% of the non-DT104 strains. The virulence of two DT104 strains was tested in orally infected day-old chicks and compared with virulence of 4 non-DT104 strains. Higher colonizing virulence of DT104 strains could be established as compared to the other strains.     

Salmonella spp. in Bavarian liquid pig manure: occurrence and relevance for the distribution of antibiotic resistance

In a representative study, 380 manure samples of pig farms distributed all over Bavaria (Germany) were screened for the presence of Salmonella spp. at the time of manure application to soil using methods adapted to DIN EN ISO 6579. The isolates were tested according to DIN 58940 for their susceptibility towards 26 antimicrobial substances - such substances partly administered in animals, but mainly used in human therapy. Six out of 380 manure samples and, in addition, the only separated liquid manure sample examined, contained Salmonella isolates. Serotypes represented S. Typhimurium (n = 5), S. Derby (n = 1) and S. Infantis/S. Serogroup C1-(6,7:r:-)-form (each n = 1). Within the serovar S. Typhimurium, the most common phage type was DT104 (n = 3). All Salmonella isolates originated from 'big' farms (>30/45 sows on combined/breeding farms or >220 fattening pigs). Four out of six manure samples contained resistant isolates; all resistant Salmonellae were multi-resistant. Two out of three DT104-isolates showed the typical penta-resistance pattern often found in S. Typhimurium DT104, which was, furthermore, supplemented by other resistances. One Escherichia coli isolated together with a phage type DT104 S. Typhimurium also expressed the same penta-resistance pattern.     

Change in antimicrobial resistance pattern in Salmonella enterica serovar Typhimurium isolates detected in a beef cattle farm

Multidrug-resistant Salmonella enterica serovar Typhimurium (S. Typhimurium) isolates with four different antimicrobial resistance patterns obtained from a beef cattle farm were characterized to determine their clonality. Macrorestriction analysis of genomic DNA revealed that these four isolates are closely related to each other and can be classified as a newly emerged pulsed-field gel electrophoresis type among cattle: cluster VII. Three of the four isolates showed resistance to extended-spectrum cephalosporins (ESCs), and this resistance was mediated by AmpC β-lactamase encoded by the bla(CMY-2) gene in a 190-kbp IncA/C plasmid. Results of restriction analysis and IncA/C backbone PCR suggest that the three 190-kbp plasmids are identical and that a 70-kbp IncA/C plasmid of the ESC-susceptible isolate is derived from the 190-kbp plasmid by a deletion event. Three isolates harboured a virulence-resistance plasmid (165 or 180 kbp), and restriction analysis revealed that these plasmids were identical or closely related to each other. These results suggest that the four S. Typhimurium cluster VII isolates originate from a common ancestor that probably invaded the farm prior to the salmonellosis outbreak. Antimicrobial resistance patterns may not necessarily reflect the relationships of the isolates.     

Abomasitis associated with multiple antibiotic resistant Salmonella enterica serotype Typhimurium phagetype DT104

Salmonella enterica serotype Typhimurium phagetype DT104 is a multiple antibiotic resistant pathogen that has been purported to be more pathogenic than other Salmonella. In this study, we evaluated the possibility that DT104 is the causative agent of veal calf abomasitis observed in four independent outbreaks of salmonellosis. This study was undertaken to determine if the outbreaks might be due to hypervirulent S. enterica serotype Typhimurium phagetype DT104 (DT104) since Salmonella does not usually cause abomasitis. Tissues and fluids from these calves were subjected to bacteriologic culture. Pure Salmonella cultures were then used in bovine challenge experiments. DT104 was identified as the causative agent of abomasitis in calves. Thus, abomasitis is a potential indicator of infection with multiple antibiotic resistant DT104 and adds credence to the apparent hypervirulence of this pathogen.     

Assessment of antibiotic resistance phenotype and integrons in Salmonella enterica serovar Typhimurium isolated from swine

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.     

Characterization of Salmonella enterica serovar Typhimurium strains of veterinary origin by molecular typing methods

Twenty-eight strains of Salmonella enterica serovar Typhimurium were characterized by three PCR-based methods. Ten strains harbored type I integrons and two different integron profiles were detected. Typing by amplified fragment length polymorphism (AFLP) resulted in observation of 10 profiles that differed by one to six bands. Salmonella strains were screened for presence of phage genes using a PCR-phage typing; five genes from P22 phage and genes encoding putative virulence factors from phages Gifsy-1, Gifsy-2 and Fels-1 were selected for testing. This set of genes was sufficient for dividing the strains into eight different PCR-phage profiles. Similar grouping of strains was observed in case of all the employed DNA techniques and they corresponded well with the phage type and antimicrobial resistance of the strains. The highest discriminating power was achieved with use of the AFLP, yet the detection of integrons and PCR-phage typing also proved to be valuable in typing the S. Typhimurium strains.     

Long-term persistence of multi-drug-resistant Salmonella enterica serovar Newport in two dairy herds

OBJECTIVE: To evaluate the association between maintaining joint hospital and maternity pens and persistence of multi-drug-resistant (MDR) Salmonella enterica serovar Newport on 2 dairy farms. DESIGN: Observational study. SAMPLE POPULATION: Feces and environmental samples from 2 dairy herds. PROCEDURE: Herds were monitored for fecal shedding of S enterica Newport after outbreaks of clinical disease. Fecal and environmental samples were collected approximately monthly from pens housing sick cows and calving cows and from pens containing lactating cows. Cattle shedding the organism were tested serially on subsequent visits to determine carrier status. One farm was resampled after initiation of interventional procedures, including separation of hospital and maternity pens. Isolates were characterized via serotyping, determination of antimicrobial resistance phenotype, detection of the CMY-2 gene, and DNA fingerprinting. RESULTS: The prevalence (32.4% and 33.3% on farms A and B, respectively) of isolating Salmonella from samples from joint hospital-maternity pens was significantly higher than the prevalence in samples from pens housing preparturient cows (0.8%, both farms) and postparturient cows on Farm B (8.8%). Multi-drug-resistant Salmonella Newport was isolated in high numbers from bedding material, feed refusals, lagoon slurry, and milk filters. One cow excreted the organism for 190 days. Interventional procedures yielded significant reductions in the prevalences of isolating the organism from fecal and environmental samples. Most isolates were of the C2 serogroup and were resistant to third-generation cephalosporins. CONCLUSIONS AND CLINICAL RELEVANCE: Management practices may be effective at reducing the persistence of MDR Salmonella spp in dairy herds, thus mitigating animal and public health risk.