S. Corti

Detection of Mycobacterium avium subspecies paratuberculosis specific IS900 insertion sequences in bulk-tank milk samples obtained from different regions throughout Switzerland

BackgroundSince Mycobacterium avium subspecies paratuberculosis (MAP) was isolated from intestinal tissue of a human patient suffering Crohns disease, a controversial discussion exists whether MAP have a role in the etiology of Crohns disease or not. Raw milk may be a potential vehicle for the transmission of MAP to human population. In a previous paper, we have demonstrated that MAP are found in raw milk samples obtained from a defined region in Switzerland. The aim of this work is to collect data about the prevalence of MAP specific IS900 insertion sequence in bulk-tank milk samples in different regions of Switzerland. Furthermore, we examined eventual correlation between the presence of MAP and the somatic cell counts, the total colony counts and the presence of Enterobacteriaceae.Results273 (19.7%) of the 1384 examined bulk-tank milk samples tested IS900 PCR-positive. The prevalence, however, in the different regions of Switzerland shows significant differences and ranged from 1.7% to 49.2%. Furthermore, there were no statistically significant (p >> 0.05) differences between the somatic cell counts and the total colony counts of PCR-positive and PCR-negative milk samples. Enterobacteriaceae occur as often in IS900 PCR-positive as in PCR-negative milk samples.ConclusionThis is the first study, which investigates the prevalence of MAP in bulk-tank milk samples all over Switzerland and infers the herd-level prevalence of MAP infection in dairy herds. The prevalence of 19.7% IS900 PCR-positive bulk-milk samples shows a wide distribution of subclinical MAP-infections in dairy stock in Switzerland. MAP can therefore often be transmitted to humans by raw milk consumption.

Bacteriological Survey of Ready-to-Eat Lettuce, Fresh-Cut Fruit, and Sprouts Collected from the Swiss Market

The consumption of ready-to-eat fresh vegetables has increased significantly in the recent decades. So far, no data are available on the bacteriological burden and the prevalence of foodborne pathogens in ready-to-eat lettuce, fresh-cut fruit, and sprouts on the Swiss market. This study was based on investigations carried out during 2 months of the summer season in 2011. Samples of 142 salads, 64 fresh-cut fruit, and 27 sprouts were included in this study. Escherichia coli, an indicator microorganism for fecal contamination, was only found in 5 lettuce samples, with amounts ranging between 2 and 3 log CFU/g. No Salmonella spp. were detected from any of the 233 samples analyzed in this study, and a low occurrence was found for contamination with L. monocytogenes, Shiga toxin-producing E. coli, enteropathogenic E. coli, and Cronobacter. From the results of the present study, we conclude that even in a country where the use of chlorine solutions to sanitize fruits and vegetables in the fresh-cut industry is not allowed, it is possible to produce ready-to-eat lettuce, fresh-cut fruit, and sprouts with high microbiological standards. Strict maintenance of good practices of hygiene at preharvest, harvest, and postharvest levels is of central importance to ensure both public health protection and product quality.

Characteristics of shiga toxin-producing Escherichia coli isolated from Swiss raw milk cheese within a 3-year monitoring program.

Food is an important vehicle for transmission of Shiga toxin-producing Escherichia coli (STEC). To assess the potential public health impact of STEC in Swiss raw milk cheese produced from cows, goats, and ewes milk, 1,422 samples from semihard or hard cheese and 80 samples from soft cheese were examined for STEC, and isolated strains were further characterized. By PCR, STEC was detected after enrichment in 5.7% of the 1,502 raw milk cheese samples collected at the producer level. STEC-positive samples comprised 76 semihard, 8 soft, and 1 hard cheese. By colony hybridization, 29 STEC strains were isolated from 24 semihard and 5 soft cheeses. Thirteen of the 24 strains typeable with O antisera belonged to the serogroups O2, O22, and O91. More than half (58.6%) of the 29 strains belonged to O:H serotypes previously isolated from humans, and STEC O22:H8, O91:H10, O91:H21, and O174:H21 have also been identified as agents of hemolytic uremic syndrome. Typing of Shiga toxin genes showed that stx(1) was only found in 2 strains, whereas 27 strains carried genes encoding for the Stx(2) group, mainly stx(2) and stx(2vh-a/b). Production of Stx(2) and Stx(2vh-a/b) subtypes might be an indicator for a severe outcome in patients. Nine strains harbored hlyA (enterohemorrhagic E. coli hemolysin), whereas none tested positive for eae (intimin). Consequently, semihard and hard raw milk cheese may be a potential source of STEC, and a notable proportion of the isolated non-O157 STEC strains belonged to serotypes or harbored Shiga toxin gene variants associated with human infections.

Incidence of Mycobacterium avium subspecies paratuberculosis in bulk-tank milk samples from different regions in Switzerland.

many animals, including primates, and is considered to be implicated in Crohns disease in human beings (HermonTaylor and others 2000). Crohns disease is a chronic granulomatous ileocolitis of humans, with significant similarity to paratuberculosis. High herd prevalences of NMAP have been found in dairy cattle in particular (Thorne and Hardin 1997). A recent USDA survey indicated that 41 per cent of dairy herds

Comparison of genomic and antimicrobial resistance features of latex agglutination test-positive and latex agglutination test-negative Staphylococcus aureus isolates causing bovine mastitis

The dairy industry suffers massive economic losses due to staphylococcal mastitis in cattle. The Staphaureux latex agglutination test (Oxoid, Basel, Switzerland) was reported to lead to negative results in 54% of bovine Staphylococcus aureus strains, and latex-negative strains are thought to be less virulent than Staphaurex latex-positive strains. However, comparative information on virulence and resistance profiles of these 2 groups of Staph. aureus is scarce. Our objective was to associate the latex agglutination phenotype of Staph. aureus strains isolated from bovine mastitis milk with data on clonal complexes, virulence genes, and antibiotic resistance to (1) determine the virulence profiles of the Staphaureux test positive and Staphaurex test negative groups, and (2) provide data needed to improve treatment of bovine mastitis and to identify potential vaccine targets. Seventy-eight Staph. aureus strains isolated from 78 cows on 57 Swiss farms were characterized. Latex agglutination was tested by Staphaureux kit, and resistance profiles were generated by disk diffusion. A DNA microarray was used to assign clonal complexes (CC) and to determine virulence and resistance gene profiles. By the Staphaureux test, 49% of the isolates were latex-positive and 51% were latex-negative. All latex-negative strains were assigned to CC151, whereas latex-positive strains were assigned to various clonal complexes, including CC97 (n=16), CC8 (n=10), CC479 (n=5), CC20 (n=4), CC7 (n=1), CC9 (n=1), and CC45 (n=1). Although the latex-negative isolates were susceptible to all antimicrobial agents tested, 24% of latex-positive isolates were classified as intermediate with regard to cefalexin-kanamycin and 13% were resistant to both ampicillin and penicillin. Microarray profiles of latex-negative isolates were highly similar, but differed largely from those of latex-positive isolates. Although the latex-negative group lacked several enterotoxin genes and sak, it exhibited significantly higher prevalence rates of genes encoding enterotoxin C, toxic shock syndrome toxin, and leukocidins (lukM/lukF-P83, lukD). Our findings suggest that latex-negative isolates represent a group of closely related strains with specific resistance and virulence gene patterns.

Clinical findings in 56 cows with toxic mastitis

The clinical, haematological, biochemical and bacteriological findings in 56 cows with toxic mastitis are described. In addition to acute mastitis, the most frequent clinical findings were tachycardia in 40 of the cows, anorexia or reduced appetite in 50, reduced ruminal and intestinal motility in 52 and no ruminal and intestinal motility in 39. The most important haematological and biochemical findings were a high haematocrit in 26 of the cows, leucopenia in 23 and a high concentration of urea in 31. In 27 of the cows the secretion of the affected mammary gland did not resemble milk. The most commonly isolated bacteria were Escherichia coli from 26 of the cows and Staphylococcus aureus from 11. Seven of the cows were investigated by an exploratory laparotomy because the principal sign was intestinal in origin, but no abnormalities were found. Seventeen of the cows were euthanased immediately after the diagnostic work, and 14 during the course of their treatment.

Arcanobacterium pluranimalium leading to a bovine mastitis: species identification by a newly developed pla gene based PCR.

We are describing a clinical case of bovine mastitis due to Arcanobacterium pluranimalium in a Holstein-Friesian heifer, delivering bloody milk on the left hindquarter. Moreover, we report on the development and evaluation of PCR primers based on the pluranimaliumlysin (pla) gene for the identification of this species. With the primer pair PlaF/PlaR the A. pluranimalium type strain as well as the mastitis isolate 704 revealed a correctly sized amplification product (458 bp), whereas no amplification product was obtained for all non-target strains. The established PCR provides a new and convenient tool for the mastitis diagnostic to differentiate between A. pluranimalium and Trueperella pyogenes.

Resistance profiles and genetic diversity of Escherichia coli strains isolated from acute bovine mastitis.

Between March 2011 and February 2012 83 E. coli strains were isolated from mastitis milk samples from 83 different animals (67 farms) and tested for their sensitivity to various antibiotics by means of disk diffusion method and genotyped by determination of the phylogenetic groups as well as by pulsed field gel electrophoresis (PFGE). The antibiotics were chosen on the basis of their licenses for intramammary application in Switzerland. As many as 16.9 % of the isolates were resistant to one or more antimicrobial agents. Amoxicillin-clavulanic acid, gentamicin and third generation cephalosporins proved effective against the majority of these strains. Nevertheless, one blaCTX-M-14 harbouring extended-spectrum-beta-lactamase producing strain was found. Genetic analysis grouped most of the strains (87 %) into phylogenetic groups A and B1. PFGE genotyping demonstrated that E. coli from cows with mastitis even from the same farm were genotypically very diverse.

Milk somatic cell count, lactate dehydrogenase activity, and immunoglobulin G concentration associated with mastitis caused by different pathogens: A field study

INTRODUCTION The aim of this study is to analyze how somatic cell counts (SCC), immunoglobulin G (IgG), and lactate dehydrogenase (LDH) interact dependent on the mastitis causing pathogen. Milk samples from 152 quarters were collected on 2 Swiss dairy farms equipped with automatic milking systems. Bacteriological culturing was performed and SCC, LDH activity and IgG concentrations were measured in each sample. Correlations and regressions among SCC, LHD, and IgG were calculated after grouping by the pathogen type (control, S. aureus, C. bovis, coagulase-negative Staphylococcus and S. uberis). All the mastitis causing pathogens were gram-positive bacteria (except for 3 cases with E. coli). In this study, the SCC and LDH were affected by the pathogen group. However, only in the S. uberis group the IgG concentration was higher than in the controls. All studied variables were positively correlated among each other. SCC and LDH were the highest correlated parameters in the control, S. aureus, C. bovis and coagulase- negative Staphylococcus groups. Only in the S. uberis group the correlation between LDH and IgG was higher than the correlation between SCC and LDH. The regression coefficients for SCC and LDH differed between groups whereas regression coefficients for SCC and IgG, and for LDH and IgG were similar in all groups. Because cases with E. coli infection were so rare, we could not include these cases in the statistical evaluation. Based on these few cases E. coli (n=3) seemed to cause a much higher increase of IgG and LDH than the infection with gram-positive bacteria. This study shows that the suitability of LDH as a marker for IgG transfer is dependent on the pathogen. The use of LDH in combination with SCC may be used as a marker to differentiate between gram-positive and gram-negative bacteria, but does not allow differentiating the immune response between different gram-positive bacteria.

Short communication: Staphylococcus aureus isolated from colostrum of dairy heifers represent a closely related group exhibiting highly homogeneous genomic and antimicrobial resistance features.

In heifers, intramammary infections caused by Staphylococcus aureus affect milk production and udder health in the first and subsequent lactations, and can lead to premature culling. Not much is known about Staph. aureus isolated from heifers and it is also unclear whether or not these strains are readily transmitted between heifers and lactating herd mates. In this study, we compared phenotypic characteristics, spa types, and DNA microarray virulence and resistance gene profiles of Staph. aureus isolates obtained from colostrum samples of dairy heifers with isolates obtained from lactating cows. Our objective was to (1) characterize Staph. aureus strains associated with mastitis in heifers and (2) determine relatedness of Staph. aureus strains from heifers and lactating cows to provide data on transmission. We analyzed colostrum samples of 501 heifers and milk samples of 68 lactating cows within the same herd, isolating 48 and 9 Staph. aureus isolates, respectively. Staphylococcus aureus strains from heifers, lactating herd mates, and an unrelated collection of 78 strains from bovine mastitis milk of mature cows were compared. With 1 exception each, characterization of all strains from heifers and lactating cows in the same herd yielded highly similar phenotypic and genotypic results. The strains were Staphaurex latex agglutination test negative (Oxoid AG, Basel, Switzerland) and belonged to agr type II, CC705, and spa types tbl 2645 and t12926. They were susceptible to all antimicrobial agents tested. In contrast, the strains from mature cows in other herds were spread across different clonal complexes, spa types, and SplitsTree clusters (http://www.splitstree.org/), thus displaying a far higher degree of heterogeneity. We conclude that strains isolated from colostrum of heifers and mastitis milk of lactating cows in the same herd feature highly similar phenotypic and genomic characteristics, suggesting persistence of the organism during the first and potentially subsequent lactations or transmission between heifers and mature herd mates.