Prevalence of antibodies to Leptospira interrogans serovar hardjoin bulk tank milk from unvaccinated irish dairy herds

: Bulk tank milk samples, collected from 347 herds throughout the Republic of Ireland using a sampling frame based on seven milk-recording organisations, were tested by ELISA for antibodies to Leptospira interrogans serovar hardjo. These herds, which had not been vaccinated against leptospirosis within the previous five years, were categorised according to their province, milk-recording organisation and size. Two-hundred-and-seventy-three herds (79%) had a positive ELISA titre. Both the probability of a herd being seropositive and the antibody level in the herd milk sample were affected by the province (P < 0.05 and P < 0.01, respectively) and the herd size category (P < 0.05 and P < 0.01, respectively). Larger herds were significantly more likely to have positive reactions and higher mean concentrations of antibody. It was concluded that a high proportion of unvaccinated Irish dairy herds have been exposed to infection with Leptospira hardjo.     

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.     

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%).     

Risk factors for clinical Salmonella enterica subsp. enterica serovar Typhimurium infection on Dutch dairy farms

Risk factors for outbreaks in 1999 of clinical Salmonella enterica subsp. enterica serovar Typhimurium infection on dairy farms were studied in a matched case-control study with 47 case farms and 47 control farms. All 47 case farms experienced a clinical outbreak of salmonellosis which was confirmed with a positive bacteriologic culture for serovar Typhimurium in one or more samples. Serovar Typhimurium phage type 401 and 506 (definitive type 104, DT104) were the most frequently isolated phage types (13 isolates). On most farms (66%), clinical signs were seen only among adult cows. The most frequently reported clinical signs were diarrhoea (in 92% of the farms) and depression (in 79% of the farms).Control farms were matched on region and had no history of salmonellosis. A questionnaire was used to collect data on case and control farms. The relationship between serovar Typhimurium status of the farm and possible risk factors was tested using conditional logistic regression. Significant factors in the final model were presence of cats on the farm (OR=0.06), purchase of manure (OR=21.5), feeding colostrum only from own dam (OR=0.08), a non-seasonal calving pattern (OR=25), unrestricted grazing of lactating cows (OR=0.07), and a high mean mowing percentage of pasture (OR=1.02).     

Nationwide survey of antibodies to bovine coronavirus in bulk milk from Swedish dairy herds.

Bulk milk samples from 2236 dairy herds randomly selected throughout Sweden in proportion to region and herd size were analysed for antibodies to bovine coronavirus (BCV) in an ELISA. The results were expressed as optical density (OD) values and an OD > 0.04 was considered positive. Eighty-nine per cent of the samples were antibody-positive and 52 per cent had high levels of antibodies to BCV (an OD > 0.70). There were significantly higher OD values (P < 0.001) and fewer antibody-negative samples (P < 0.001) from larger herds than from smaller herds. There were also significantly higher OD values and fewer antibody-negative samples from herds in southern Sweden than from herds in northern Sweden (P < 0.001 and P < 0.001, respectively). These results indicate a higher frequency of BCV infections in larger herds and in herds in southern Sweden.     

Comparison of Salmonella enterica subspecies enterica serovar Typhimurium isolates from dairy cattle and humans using in vitro assays of virulence

Salmonella enterica subspecies enterica serovar Typhimurium (Salmonella typhimurium) can infect and cause disease in a wide range of host species however there have been suggestions that this serovar may have genes involved with host range or specificity [Tsolis, R.M., Townsend, S.M., Miao, E.A., Miller, S.I., Ficht, T.A., Adams, L.G., Baumler, A.J., 1999. Identification of a putative S. enterica serotype Typhimurium host range factor with homology to IpaH and YopM by signature-tagged mutagenesis. Infect. Immun. 67 (12), 6385-6393]. Our goal in this study was to determine if in vitro virulence assays would support this suggestion. Twelve human and 10 bovine isolates of S. typhimurium from a single county in California were evaluated using in vitro virulence assays of adhesion and invasion. The resulting data was combined with results from previously reported genotypic and phenotypic testing of the isolates and statistical analysis performed using multivariate general linear models. Human isolates had higher adhesion values in each of the statistical models tested (p<0.05) but no statistical differences were found in the invasion values of human and bovine source isolates. Both adhesion and invasion values differed between the two largest groups of isolates segregated on the basis of pulsed-field gel patterns. The findings suggest there may be genetically defined in vitro virulence attributes in S. typhimurium that are associated with host species.     

Prevalence of Leptospira interrogans serovar hardjo antibodies in milk in Belgian dairy herds

Pooled milk samples were collected from 2000 Belgian dairy herds in the autumn of 1989. Antibodies against Leptospira interrogans serovar hardjo were detected in 9.2% of the herds, with the incidence being higher in the southern part of the country.     

Risk factors for Coxiella burnetii antibodies in bulk tank milk from Danish dairy herds

The aim was to identify risk factors associated with Coxiella burnetii antibody positivity in bulk tank milk (BTM) samples from 100 randomly selected Danish dairy cattle herds. Antibody levels were measured by an enzyme-linked immuno-sorbent assay. Before testing the herds, the farm managers were interviewed about hired labour, biosecurity, housing and herd health during the 12 months prior to the study. Variables considered important for C. burnetii antibody positivity in multivariable logistic regression analysis included the sharing of machines between farms (OR = 3.6), human contacts (OR = 4.2), artificial insemination by other people than artificial insemination technicians (OR = 7.7), routine herd health contract with the veterinarian (OR = 4.3) and hygiene precautions taken by veterinarians (OR = 5). In addition, herd size, hired labour, trading of cattle between farms, quarantine and use of calving and disease pens also showed significant association in univariable analysis. This study demonstrates that strict biosecurity is important for the prevention of infections with C. burnetii.     

Mitsuokella jalaludinii inhibits growth of Salmonella enterica serovar Typhimurium

Salmonella continues to be a significant human health threat, and the objective of this study was to identify microorganisms with the potential to improve porcine food-safety through their antagonism of Salmonella. Anaerobic culture supernatants of 973 bacterial isolates from the gastrointestinal tract and feces of swine were screened for their capacity to inhibit the growth of Salmonella enterica serovar Typhimurium. Growth inhibition of 1000-fold or greater was observed from 16 isolates, and 16S rRNA sequencing identified the isolates as members of the genera Mitsuokella, Escherichia/Shigella, Anaerovibrio, Selenomonas, and Streptococcus. Four isolates were identified as Mitsuokella jalaludinii, and the mechanism of Salmonella Typhimurium growth inhibition by M. jalaludinii was further investigated. M. jalaludinii stationary phase culture supernatants were observed to significantly inhibit growth, and featured the production of lactic, succinic, and acetic acids. Aerobic and anaerobic S. Typhimurium growth was restored when the pH of the culture supernatants (pH 4.6) was increased to pH 6.8. However, S. Typhimurium growth in fermentation acid-free media was the same at pH 4.6 and pH 6.8 - indicating a synergistic effect between fermentation acid production and low pH as the cause of S. Typhimurium growth inhibition. Furthermore, exposure of S. Typhimurium to M. jalaludinii culture supernatants inhibited Salmonella invasion of HEp-2 cells by 10-fold. The results identify M. jalaludinii as a possible inhibitor of Salmonella growth and invasion in swine, and thus a potential probiotic capable of improving food safety.     

Antimicrobial resistance in Salmonella enterica serovar Typhimurium from human and animal sources in Italy

Salmonella Typhimurium strains isolated in Italy in the period 2002-2004 from human and animal sources were examined for their antimicrobial susceptibility. Resistance to tetracycline (T, 73.6%), sulfonamides (Su, 73.3%), ampicillin (A, 67.6%), streptomycin (S, 65.4%) and chloramphenicol (C, 32.3%) were frequently observed. Resistance to ciprofloxacin was only observed in a swine strain, but most human strains resistant to nalidixic acid showed reduced susceptibility to that drug (MIC > or = 0.125 mg/l). Overall, 64% of the strains were resistant to four or more drugs. The most common resistance profiles were ACSSuT, prevalent in strains belonging phage type DT104 and ASSuT, prevalently associated with strains unable to be typed.     

Testing of bulk tank milk for Salmonella Dublin infection in Danish dairy herds.

The usefulness of enzyme-linked immunosorbent assay (ELISA) was investigated as a simple method to screen for Salmonella Dublin infection in dairy herds, examining bulk tank milk samples for lipopolysaccharide (O:1,9,12) antibodies. The cut-off value for the ELISA on bulk tank milk was established based on individual milk samples (n = 2887) and bulk tank milk from 52 herds. Bulk tank milk samples (n = 5108) were collected from 1464 dairy herds located in 19 different areas. About 10% of the dairy herds in Denmark participated in the study. The percentage of herds changing from test-negative to test-positive in each area was correlated with the incidence of S. Dublin outbreaks in the corresponding county (r = 0.48, n = 19; P < 0.025). The mean level of the OD values obtained in the first and third test rounds was not constant (Pr /t/ = 0.0001). The study demonstrated that the probability of being test-negative in the third test round was 0.926 for a herd with 2 previous test-negative results. It was concluded that the investigated ELISA method was in general accordance with the cases of clinical S. Dublin infection recorded, and that the method has a potential for national screening purposes.     

A survey on Dictyocaulus viviparus antibodies in bulk milk of dairy herds in Northern Germany

Parasitic bronchitis caused by the bovine lungworm, Dictyocaulus viviparus, occurs worldwide in temperate areas. The parasite is found predominantly in calves and heifers, but dairy cattle can suffer from lungworms when they become infected for the first time or if they have lost immunity due to lack of exposure to lungworm larvae during the grazing season. The present study was performed to determine the D. viviparus bulk milk antibody prevalence in dairy herds in the East Frisian region of northwestern Germany, Lower Saxony, by analysing bulk milk samples collected in January (860 samples), September (866 samples) and November (860 samples) 2008, thereby representing 906 dairy farms. These samples were tested for antibodies against D. viviparus by a milk ELISA. This test detects patent infections only since it is based on recombinant major sperm protein as antigen. While in January 12.8% of dairy farms were positive for D. viviparus antibodies, the bulk milk samples collected in September and November revealed 6.9% and 6.6% positive dairy herds. From the 906 dairy farms included in the study, 191 (21.1%) tested positive at least once for antibodies against lungworm. From 810 dairy farms from which bulk milk samples were obtained during all three samplings, 146 (18.0%) farms were positive at one sampling date, 27 (3.3%) at two, and 4 (0.5%) on all three sampling dates. The majority of the farms represented in the study belonged to four districts of East Frisia, which showed no significant difference in the proportion of positive dairy farms.     

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.     

Antimicrobial resistance in Salmonella enterica serovar Dublin isolates from beef and dairy sources

Salmonella enterica serovar Dublin (S. Dublin) is a cattle-adapted Salmonella serovar, so if antimicrobial resistance in S. Dublin arises as a result of antimicrobial use this most likely occurs within the cattle reservoir without impact from antimicrobial use in humans. We tested the antimicrobial resistance of bovine-origin S. Dublin isolates from 1986 through 2004 using a standard disk diffusion method. High proportions of isolates throughout the time period were resistant to one or more antimicrobials, and a marked increase in resistance to ceftazidime occurred between 2000 and 2004. Dairy-origin isolates were more likely to be resistant to several antibiotics than were isolates from beef operations where exposure to antimicrobials is likely to be less frequent. Plasmid analysis of a subset of isolates also supported the hypothesis that antimicrobial resistance traits in the cattle-adapted serovar Dublin were acquired within the bovine host environment.     

Phage Types of Salmonella enterica ssp. enterica serovar Typhimurium Isolated from Production Animals and Humans i Denmark

S. Typhimurium is one of the 2 most common salmonella serotypes causing human salmonellosis in Denmark. In order to illustrate the significance of different production animals as a source of infection, 1461 isolates were characterized by phage typing. The isolates originated from human patients and from cattle, pigs and poultry. By phage typing the isolates could be separated in 35 different phage types. Five types (10, 12, 66, 110 and 135) predominated and comprised 78.8% of the isolates. In humans, 57.3% of the isolates were phage type 12. This phage type was also predominant in pig herds and, to a lesser degree, in cattle. Phage types 110, 120, 135 and 193 constituted 86.5% of the poultry isolates while these phage types only made up 12.9% of the human isolates. The investigation showed that pigs are probably a major source of S. Typhimurium infection in humans in Denmark today.     

Pulsed field gel electrophoresis for animal Salmonella enterica serovar Typhimurium isolates in Taiwan

Salmonella enterica subspecies enterica serovar Typhimurium is a common pathogen for humans and animals. In order to trace the clonal relationship and to find the circulating strains between human and animal isolates, chromosomal DNAs from 87 serovar Typhimurium strains isolated from animals (pigs were the majority) were subjected to XbaI and SpeI digestion and pulsed field gel electrophoresis (PFGE). For the 87 animal isolates, 38 PFGE pattern combinations were obtained. As the subtyping results from animal isolates were compared with those from the 45 human isolates, it was found that 14 of the animal isolates and 13 of the human isolates shared a common PFGE pattern combination, i.e., pattern XgSf (or called X5S4). When these human and animal isolates were subjected to antibiotic susceptibility test using 11 antibiotics, it was found that strains of pattern XgSf (X5S4) belong to a common antibiogram pattern which is tetracycline, gentamicin, ampicillin, streptomycin and chloramphenicol resistant. Since most of the animal and human strains in pattern XgSf were originally isolated from various areas over different years, strains of this PFGE pattern may be the most epidemic strains which circulating between human and animal sources.     

Apparent prevalence of antibodies to Coxiella burnetii (Q fever) in bulk tank milk from dairy herds in southern Belgium

In 2006, a cross-sectional survey was carried out in 206 herds in southern Belgium to estimate the proportion of herds with Q fever-seropositive bulk tank milk (BTM), i.e. herds with an intra-herd seroprevalence >10%. Of the tested herds, 119 (57.8%; 95% CI: 51.1-64.4%) were seropositive. Most had a low antibody titre. Of these herds, 50 were randomly subjected to a real time Coxiella burnetii PCR analysis. The proportion of herds excreting Coxiella in BTM was 30.0% with only 2.0% of herds having a high level of shedding. An additional questionnaire and logistical regression identified some factors which increased the risk of BTM seroconversion (in heifers, drinking water from watercourse or well water) and some which decreased it (in heifers, tie- and free-stalling, and animals of all age in the same shed; in heifers and cows, shed disinfection).     

Early epithelial invasion by Salmonella enterica serovar Typhimurium DT104 in the swine ileum

Salmonella enterica serovar Typhimurium is an important intestinal pathogen in swine. This study was performed to document the early cellular invasion of Salmonella serovar Typhimurium in swine ileum. Ileal gut-loops were surgically prepared in ten 4- to 5-week-old mixed-breed pigs and inoculated for 0-60 minutes. Loops were harvested and prepared for both scanning and transmission electron microscopy (SEM and TEM, respectively). Preferential bacterial adherence to microfold cells (M cells) was seen within 5 minutes, and by 10 minutes bacterial invasion of the apical membrane was seen in M cells, goblet cells, and enterocytes. This multicellular invasion was observed throughout the course of infection. In addition, SEM revealed a specific affinity of Salmonella serovar Typhimurium to sites of cell extrusion. Using TEM, bacteria in these areas were focused in the crevices formed by the extruding cell and the adjacent cells and in the cytoplasm immediately beneath the extruding cell. Our results suggest that early cellular invasion by Salmonella serovar Typhimurium is nonspecific and rapid in swine. Furthermore, the combination of SEM and TEM data suggests that Salmonella serovar Typhimurium may use sites of cell extrusion as an additional mechanism for early invasion.