Biofilm formation of Staphylococcus aureus dairy isolates representing different genotypes.

The objective of this study was to compare the biofilm-forming capabilities of different genotypes of Staphylococcus aureus dairy isolates from Switzerland and northern Italy, including Staph. aureus genotype B (GTB) and methicillin-resistant Staph.aureus (MRSA). We hypothesized that biofilm formation might be more pronounced in the contagious GTB isolates compared with other genotypes affecting individual animals. Twenty-four dairy isolates, including 9 MRSA, were further characterized by genotyping by using ribosomal spacer PCR, spa typing, biofilm formation under static and dynamic conditions, and scanning electron microscopy. The GTB isolates (n = 6) were more able to form biofilms than other genotypes at 37°C and at 20°C after 48 and 72 h of incubation in the static assay using polystyrene microtiter plates. This result was supported by scanning electron micrographs showing a GTB isolate producing strong biofilm with extracellular matrix in contrast to a genotype C isolate. Furthermore, none of the MRSA isolates formed strong biofilms in the static assay. However, some MRSA produced low or moderate amounts of biofilm depending on the applied conditions. Under dynamic conditions, a much more diverse situation was observed. The ability of GTB isolates to be strong biofilm formers was not observed in all cases, emphasizing the importance of growth conditions for the expression of biofilm-related genes. No specific genotype, spa type, or MRSA isolate could be categorized significantly into one level of biofilm formation. Nineteen percent of isolates behaved similarly under static and dynamic conditions. The results of this study expand our knowledge of different dairy-related Staph. aureus subtypes and indicate the benefit of genotyping when biofilms are studied.

Antimicrobial resistance pattern of methicillin-resistant Staphylococcus aureus in the food industry.

There is increasing concern about the impact on public health of methicillin-resistant Staphylococcus aureus (MRSA) associated with animal food products. MRSA remains a serious problem because of the high incidence and multidrug resistance of the strains, even for strains isolated from foods, food environments and food handlers. The objectives of this study are: (i) to evaluate the susceptibility of S. aureus strains isolated from food, food handlers and food-processing environments to 14 antibiotics currently used in veterinary and human therapy; (ii) to assess the presence of the mecA gene. A total of 1007 samples were collected from food, food handlers, and environments and were analyzed for the presence of S. aureus. S. aureus was present in 165 of the 1007 samples. A total of 157 isolates were methicillin-susceptible S. aureus (MSSA) and 8 isolates were MRSA. In particular, out of 8 MRSA strains detected, 4 strains harboured the mecA gene. All MRSA strains were resistant to at least one of the tested antibiotics and 6 strains demonstrated multi-resistance. Considering the high level of resistances in S. aureus and the isolation of MRSA strains, the surveillance of antimicrobial resistance and the spreading of this pathogen is of crucial importance in the food production chain. These data are useful in improving background data on antimicrobial resistance of S. aureus isolated from food, processing environments and food handlers, supporting the prudent use of antibiotics and the development of international control programs.

Distribution of aminogenic activity among potential autochthonous starter cultures for dry fermented sausages.

Any bacterial strain to be used as starter culture should have suitable characteristics, including a lack of amino acid decarboxylase activity. In this study, the decarboxylase activity of 76 bacterial strains, including lactic acid bacteria and gram-positive, catalase-positive cocci, was investigated. These strains were previously isolated from European traditional fermented sausages to develop autochthonous starter cultures. Of all the strains tested, 48% of the lactic acid bacteria strains and 13% of gram-positive, catalase-positive cocci decarboxylated one or more amino acids. Aminogenic potential was strain dependent, although some species had a higher proportion of aminogenic strains than did others. Thus, all Lactobacillus curvatus strains and 70% of Lactobacillus brevis strains had the capacity to produce tyramine and beta-phenylethylamine. Some strains also produced other aromatic amines, such as tryptamine and the diamines putrescine and cadaverine. All the enterococcal strains tested were decarboxylase positive, producing high amounts of tyramine and considerable amounts of beta-phenylethylamine. None of the staphylococcal strains had tyrosine-decarboxylase activity, but some produced other amines. From the aminogenic point of view, Lactobacillus plantarum, Lactobacillus sakei, and Staphylococcus xylosus strains would be the most suitable for use as autochthonous starter cultures for traditional fermented sausages.

Influence of temperature on biofilm formation by Listeria monocytogenes on various food-contact surfaces: relationship with motility and cell surface hydrophobicity.

AIMS: To assess the ability of Listeria monocytogenes to form biofilm on different food-contact surfaces with regard to different temperatures, cellular hydrophobicity and motility. METHODS AND RESULTS: Forty-four L. monocytogenes strains from food and food environment were tested for biofilm formation by crystal violet staining. Biofilm levels were significantly higher on glass at 4, 12 and 22 degrees C, as compared with polystyrene and stainless steel. At 37 degrees C, L. monocytogenes produced biofilm at significantly higher levels on glass and stainless steel, as compared with polystyrene. Hydrophobicity was significantly (P < 0.05) higher at 37 degrees C than at 4, 12 and 22 degrees C. Thirty (68.2%) of 44 strains tested showed swimming at 22 degrees C and 4 (9.1%) of those were also motile at 12 degrees C. No correlation was observed between swimming and biofilm production. CONCLUSIONS: L. monocytogenes can adhere to and form biofilms on food-processing surfaces. Biofilm formation is significantly influenced by temperature, probably modifying cell surface hydrophobicity. SIGNIFICANCE AND IMPACTS OF THE STUDY: Biofilm formation creates major problems in the food industry because it may represent an important source of food contamination. Our results are therefore important in finding ways to prevent contamination because they contribute to a better understanding on how L. monocytogenes can establish biofilms in food industry and therefore survive in the processing environment.

Traditional dry fermented sausages produced in small-scale processing units in Mediterranean countries and Slovakia. 1: Microbial ecosystems of processing environments.

Microbial ecosystems were surveyed in 314 environmental samples from 54 Southern and Eastern European small-scale processing units (PUs) manufacturing traditional dry fermented sausages. The residual microflora contaminating the surfaces and the equipment were analysed after cleaning and disinfection procedures. All the PU environments were colonised at various levels by spoilage and technological microflora with excessive contamination levels in some of the PUs. Sporadic contamination by pathogenic microflora was recorded. Salmonella and Listeria monocytogenes were detected in 4.8% and 6.7% of the samples, respectively, and Staphylococcus aureus was enumerated in 6.1% of the samples. Several critical points were identified, such as the machines for S. aureus and the tables and the knives for L. monocytogenes; this knowledge is crucial for the improvement of hygiene control systems in small and traditional meat processing industries. The variability of the residual contamination emphasized the different cleaning, disinfecting and manufacturing practices routinely followed by these small-scale processing units.

Antimicrobial susceptibility of environmental Staphylococcus aureus strains isolated from a pigeon slaughterhouse in Italy.

No information is available concerning the antimicrobial susceptibility of Staphylococcus aureus isolated from pigeon slaughterhouses. In the present study, 59 staphylococcal strains isolated from a pigeon slaughterhouse in central Italy were compared according to their antibiotic resistance. On the basis of cultural and biochemical properties, all isolates could be identified as S. aureus. The strains were checked for the productions of enterotoxins A, B, C, D by reversed passive latex agglutination. Resistance to 26 antibiotics was also determined paying particular attention to resistance to those antimicrobial agents frequently used in human medicine and in poultry breeding. Only one strain was positive for the production of enterotoxins type C and D. It was isolated from the evisceration tube after slaughtering. Enterotoxin B was produced by 2 strains isolated from the eyebrows and conjunctivas of the worker operating the crop rinsing tube. As to the susceptibility to antibiotics, all strains were sensitive to amoxicillin/clavulanic acid, bacitracin, cephalothin, fusidic acid, gentamicin, kanamycin, linezolid, oxacillin, quinupristin/dalfopristin, rifampicin, tobramycin, trimethoprim-sulfamethoxazole, vancomycin. Some (15.2%) of the strains were resistant to ampicillin and to penicillin G; 6.8% were resistant to chloramphenicol, 20.3% to enrofloxacin, 16.9% to erythromycin and to ciprofloxacin, 8.5% to clindamycin, and 11.9% to lincomycin. The highest percentages of strains were resistant to tetracycline and oleandomicin (37.3 and 25.4% respectively). Methicillin-resistant staphylococci were also found (3.4%). Only one strain had a multiple antibiotic resistance index > 0.30. The results were statistically analyzed and clustered in 6 groups. This work provides the antibiotic resistance pattern of S. aureus strains isolated from a pigeon slaughtering plant and represents a study on a quite unknown field in meat production.

The challenge of education for the veterinary inspector.

The aim of this review is to provide an insight into the current state of, and future changes in, veterinary education. At present, Veterinary Inspectors need to have the appropriate education, relevant experience and the ability to carry out their duties in the context of inspections. They should respond to the changes in legislation related to food and to the official control systems and must take into account the Community rules. In effect, there is still a wide diversity among schools in Italy and in Europe, showing that there is an urgent need for harmonization of the veterinary education, in order to have a common basis of knowledge. Moreover, it is important to maintain and update all knowledge by means of the so-called lifelong learning. Along the educational route of the veterinary inspector, many gaps still exist: the entire system of learning should be revised, with the aim of having a well-integrated education system in cooperation with all the figures involved. In this way Veterinary Inspectors will be able to face the new issues posed by society and the globalization of food consumption and will be able to maintain their position in the assurance of food safety.

Relationship between immune system and gram-negative bacteria, binding of Salmonella pullorum-gallinarum to chicken lymphocytes.

Salmonella pullorum-gallinarum binding to chicken lymphocytes has been evaluated in individuals of different age (1-20 wks). Bacterial adherence has been studied on lymphoid cells recovered from thymus, spleen, bursa and peripheral blood. Binding was age-dependent and neuraminidase treatment of lymphoid cells led to an increase of adherence. In addition, by using two monoclonal antibodies, the CT-1, which defines thymocytes and the M-4 which identifies surface IgM and an avian homologue of mammalian IgD on B lymphocytes, distribution of Salmonella pullorum-gallinarum binding to T and B lymphocytes has been analyzed. Results show that either T lymphocytes or B lymphocytes have the capacity to form rosettes with Salmonella. In particular, B cells from bursa and spleen display the highest capacity of adherence to bacteria, this supporting the defence function which has been attributed to bursa of Fabricius.