Use of plasmid profiles to detect changes in strains of Staphylococcus aureus during poultry processing

The plasmid profiles of Staphylococcus aureus strains isolated at different stages in three poultry processing plants have been examined. Changes in profiles were seen in two plants after the plucking stage and the appearance of these new profiles correlated with the presence of an endemic strain, as suggested previously by increases in bacterial counts and changes in biotypes at the same stage. A third plant in which such changes did not occur showed no change in profiles. Plasmid profiles are therefore a rapid and sensitive method for distinguishing endemic strains within a plant from the flora of the incoming birds. Certain profiles also appeared to correspond with particular biotypes and certain phage types.     

Chlorine resistance of strains of Staphylococcus aureus isolated from poultry processing plants

Strains of Staphylococcus aureus which colonize defeathering machinery and become endemic within poultry processing plants appear to be resistant to the normal cleaning and disinfection processes. The resistance of endemic strains to chlorine was compared with that of isolates from the natural skin flora of poultry. Endemic strains were almost eight times more resistant and this was due primarily to their ability to grow in macroclumps but also to the production of an extracellular slime layer.     

Plasmid profiles as indicators of the source of contamination of Staphylococcus aureus endemic within poultry processing plants

A total of 530 strains of Staphylococcus aureus were isolated from the defeathering machinery of a chicken processing plant and from neck skin samples of carcasses at different stages of processing in two visits 4 weeks apart. Eleven different plasmid profiles were detected in the isolates, eight being common to both visits. The plasmid profiles of the strains forming the majority of the population on the freshly slaughtered birds were rarely present in the strains isolated from the pluckers (except at the entry to the first plucker) and were present in only a small proportion of the strains isolated from carcasses after plucking. However, the profiles from the strains isolated from the pluckers on both visits were different from those forming the majority of the population on the incoming birds but formed the major part of the carcass flora after plucking, suggesting that such strains were endemic. These strains were found as a small proportion of the isolates made from the incoming birds, suggesting that this was the route by which the endemic strains were introduced into the plant. Such endemic strains exhibited a clumping growth, even in liquid shake culture, which may have made it easier for them to become established on the pluckers and to resist cleaning and disinfection. This clumping phenotype was correlated with the presence of a 7.5-megadalton plasmid.     

Differentiation of Staphylococcus aureus from freshly slaughtered poultry and strains 'endemic' to processing plants by biochemical and physiological tests

A comparison was made of 27 'endemic' strains of Staphylococcus aureus and 35 strains from freshly slaughtered birds, isolated at five commercial slaughterhouses processing chickens or turkeys. Of 112 biochemical and physiological tests used, 74 gave results which differed among the strains. Cluster analysis revealed several distinct groupings which were influenced by strain type, processing plant and bird origin; these included a single group at the 72% level of similarity containing most of the 'endemic' strains. In comparison with strains from freshly slaughtered birds, a higher proportion of 'endemic' strains produced fibrinolysin, alpha-glucosidase and urease and were beta-haemolytic on sheep-blood agar. The 'endemic' type also showed a greater tendency to coagulate human but not bovine plasma, and to produce mucoid growth and clumping. The last two properties, relevant to colonization of processing equipment, were less evident in heart infusion broth than in richer media or process water collected during defeathering of the birds.     

Plasmid profiles as indicators of the source of contamination of Staphylococcus aureus endemic within poultry processing plants.

A total of 530 strains of Staphylococcus aureus were isolated from the defeathering machinery of a chicken processing plant and from neck skin samples of carcasses at different stages of processing in two visits 4 weeks apart. Eleven different plasmid profiles were detected in the isolates, eight being common to both visits. The plasmid profiles of the strains forming the majority of the population on the freshly slaughtered birds were rarely present in the strains isolated from the pluckers (except at the entry to the first plucker) and were present in only a small proportion of the strains isolated from carcasses after plucking. However, the profiles from the strains isolated from the pluckers on both visits were different from those forming the majority of the population on the incoming birds but formed the major part of the carcass flora after plucking, suggesting that such strains were endemic. These strains were found as a small proportion of the isolates made from the incoming birds, suggesting that this was the route by which the endemic strains were introduced into the plant. Such endemic strains exhibited a clumping growth, even in liquid shake culture, which may have made it easier for them to become established on the pluckers and to resist cleaning and disinfection. This clumping phenotype was correlated with the presence of a 7.5-megadalton plasmid.     

Differentiation of Staphylococcus aureus from freshly slaughtered poultry and strains 'endemic'to processing plants by biochemical and physiological tests

A comparison was made of 27 'endemic' strains of Staphylococcus aureus and 35 strains from freshly slaughtered birds, isolated at five commercial slaughterhouses processing chickens or turkeys. Of 112 biochemical and physiological tests used, 74 gave results which differed among the strains. Cluster analysis revealed several distinct groupings which were influenced by strain type, processing plant and bird origin; these included a single group at the 72% level of similarity containing most of the 'endemic' strains. In comparison with strains from freshly slaughtered birds, a higher proportion of 'endemic' strains produced fibrinolysin, aP-glucosidase and urease and were β-haemolytic on sheep-blood agar. The 'endemic' type also showed a greater tendency to coagulate human but not bovine plasma, and to produce mucoid growth and clumping. The last two properties, relevant to colonization of processing equipment, were less evident in heart infusion broth than in richer media or process water collected during defeathering of the birds.     

Destruction of Staphylococcus aureus during frankfurter processing.

We studied the thermal resistance of Staphylococcus aureus during frankfurter processing in respect to whether staphylococci are killed by the heating step of the process and whether heat injury interferes with the quantitative estimation of the survivors. With S. aureus 198E, heat injury could be demonstrated only when large numbers of cells (10(8)/g) were present and at a product temperature of 140 degrees F (60 degrees C). On tryptic soy agar and tryptic soy agar plus 7% NaCl media, at temperatures less than 140 degrees F, the counts were virtually identical; above 140 degrees F, the counts converged, with the organisms dying so rapidly that heat injury was not demonstrable. Heat injury was thus judged not to interfere with the quantitative estimation of staphylococci surviving the normal commercial heating given frankfurters. By using a combination of direct plating on tryptic soy agar and a most-probable-number technique, we detected no viable cells (less than 0.3/g) of several strains of S. aureus in frankfurters heated to 160 degrees F (71.1 degrees C). This temperature is compatible with the normal final temperature to which federally inspected processors heat their frankfurters and with the temperature needed to destroy salmonellae.     

Enterotoxigenicity of Staphylococcus aureus strains isolated from poultry: raw poultry carcases as a potential food-poisoning hazard

Staphylococcus aureus strains were isolated from end-of-lay poultry carcases obtained from a plant at two different stages of processing before and after storage at different temperatures. These strains were supplemented with Staph. aureus strains isolated from poultry from a wide range of sources and biotyped, phage typed, and tested for production of enterotoxins A-E. The isolates were found to consist of poultry and human specific strains and each of these groups contained strains able to produce enterotoxin. Poultry strains produced only enterotoxin D whereas human strains produced enterotoxins A, C and D. The hen carcases used in storage experiments were found to be naturally contaminated with enterotoxin D producing staphylococci. No enterotoxin D could be detected on any of the carcases even after storage at temperatures which allowed multiplication of the organisms to occur (final Staph. aureus counts ranged from 10² to 10⁷/16 cm² of breast skin).     

Decreased resistance to antibiotics and plasmid loss in plasmid-carrying strains of Staphylococcus aureus treated with ascorbic acid

The effect of ascorbic acid on plasmid-coded antibiotic resistance in Staphylococcus aureus was investigated. Several strains of S. aureus were cultured in the presence of 1 mM ascorbate for 6 h. This treatment induced an increased loss of resistance markers in 4 of 6 strains tested, and agarose gel electrophoresis showed this disappearance of plasmid DNA in ascorbate-induced susceptible colonies. The presence of ascorbate induced a 50-75% decrease in minimal inhibitory concentrations of different antibiotics for resistant strains. When ascorbate is added, formerly subinhibitory concentrations of penicillin or tetracycline have an increased inhibitory effect on resistant strains and even induced the death of 25-93% of the initial population. These results suggest that ascorbate can induce the loss of several plasmids of S. aureus, and that the levels of antibiotic resistance are also affected by the presence of this compound.     

Prophage-dependent plasmid integration in Staphylococcus aureus.

A study has been done of reversion to thermostability of thermosensitive, replication-defective (TSR) mutant penicillinase plasmids. All three of the expected classes of reversions were encountered: back mutation, suppression, and integration. The latter class was examined in some detail and it was found that the presence of the phi 11 phophage enhance the frequency of reversion by integration some 103-fold. Prophage-dependent integration resulted in inactivation of plasmid-linked arsenate and arsenite resistance; these revertant strains gave rise to high frequency tranducing lysates where the plasmid was restored upon transduction to its original TSR state including recovery of these resistances. The integrated plasmid-prophage complexes were stable at high temperatures (43 C) but slow growing and unstable at low (32 C); loss of either plasmid or prophage restored normal growth and stability. Sometimes restoration of the plasmid to its autonomous TSR state was observed and molecular studies showed that in most cases the plasmid was essentially the same size as before integration. In some cases an excision complex was recovered that was more than twice the size of the plasmid and could have been a plasmid-phage co-integrate. Integration also took place in the absence of the ? 11 prophage. These integrations retained all plasmid-linked resistances, were stable at all temperatures, and gave rise to low frequency transducing lysates in which the integrated state was retained upon transduction. On the basis of these results it is suggested that the prophage promotes integration at or near its attachment site.     

Molecular diversity and characterization of tetracycline-resistant Staphylococcus aureus isolates from a poultry processing plant

DNA fingerprinting and molecular characterization showed that the tetracycline-resistant Staphylococcus aureus population of a South African poultry processing plant comprised one or possibly several tet(K)-containing endemic clones that contaminated chicken and machinery surfaces at all sampled processing stages. The tet(K) gene was transferable by filter mating to S. aureus recipient 80CR5 and was located on a pT181-like plasmid.     

Antimicrobial resistance and toxin gene profiles of Staphylococcus aureus strains from Holstein milk

Isolation of Staphylococcus aureus (Staph. aureus) from Holstein milk samples with mastitis and nonmastitis was conducted to estimate its prevalence, antimicrobial resistance and toxin genes. A total of 353 milk samples were collected from three Chinese Holstein herds. Fifty‐three Staph. aureus isolates collected from 29 Staph. aureus‐positive samples were characterized via antimicrobial susceptibility, toxin genes and Pulsed‐field Gel Electrophoresis (PFGE) profiles. The prevalence of Staph. aureus was 4·0–9·5% in mastitic and 7·3–11·5% in nonmastitic samples in the analysed herds. Approximately 61·0% of Staph. aureus strains isolated from mastitis cows were resistant to ≥10 antimicrobials compared with 0% of isolates with nonmastitis. The most frequently observed super antigenic toxin gene was pvl (41·5%) followed by seh + pvl (13·2%). We did not find mecA‐positive methicillin‐resistant Staph. aureus (MRSA) strains, while mecA‐negative MRSA strains were identified in the three herds. PFGE results suggested potential transmission of Staph. aureus strains in different farms. These results open new insights into Staph. aureus transmission and antimicrobial resistance of Holstein dairy cows and into developing strategies for udder health improvement of dairy cattle.     

Detection of low-enterotoxin-producing Staphylococcus aureus strains

Culture supernatant fluids from 26 (23.6%) monkey feeding test-positive Staphylococcus aureus strains, negative for enterotoxins by gel diffusion, were positive by enzyme-linked immunosorbent assay for one or more of the identified enterotoxins. Staphylococcal enterotoxin D (SED) was produced by 23 (88.5%) strains, SED and SEA were produced in two strains, and SED and SEC were produced in one strain. One strain produced only SEA, and two strains produced only SEC.     

Nonspecific adhesion of Staphylococcus aureus strains to solid surfaces

Autoagregating strains of bacteria are characterised by high surface hydrophobicity, which determines their ability to adhesion. An assessment was done of non-specific adhesion to solid surfaces of S. aureus strains isolated from blood, pus and nasopharynx of hospitalised people. The method used made possible differentiation of strains, which were studied, on the basis of their surface characteristics. Their properties decide about the abilities of strains to the colonisation of host tissues and at the same time they influence their potential virulence. In the study attention was also paid to the participation of surface proteins in the processes of adhesion cells to glass surfaces.