Biochemical Characteristics and Enterotoxigenicity of Staphylococcus aureus Strains Isolated from Poultry

About half (49%) of strains of Staphylococcus aureus isolated from poultry were non-typable with the international human set of phages, and 55% were biotype B according to the biochemical identification scheme of Hájek & Maršálek (1971, 1973). A furthest neighbour clustering strategy and principal coordinate analysis based on 17 biochemical tests made clear distinctions between biotype B strains and a group of biotype A and intermediate strains. Overall 62% of strains were enterotoxigenic, the majority producing enterotoxin A. Significantly fewer intermediate strains than biotype A or B strains were enterotoxigenic. Starch gel zymograms of intracellular esterases showed a general correlation with the biotyping and phage typing results.     

Dialysis Culture for the Production of Extracellular Protein A from Staphylococcus aureus A676

Different nutrient media have been tested for the production of extracellular protein A from Staphylococcus aureus A676. High yields were obtained in trypticase soy broth with glucose as the main energy source. In aerated fermenter cultures the amount of protein A obtained was 0·2–0·3 gl⁻¹, as compared to reported yields of 0·03 gl⁻¹. Yields were further increased in dialysis cultures, where protein A concentrations of 2 gl⁻¹. were achieved.     

The Incidence of Enterotoxin Production in Strains of Staphylococcus aureus Isolated from Foods

S ummary . The production of staphylococcal enterotoxins A, B, C, D and E among 200 strains of Staphylococcus aureus was surveyed using a double diffusion immunoprecipitation technique. Enterotoxin A was found to occur most frequently and enterotoxin B least frequently. The distribution of enterotoxigenicity in strains isolated from meat products differed from that of strains isolated from dairy products. The correlations of porcine plasma coagulation and of bacteriophage pattern with enterotoxigenicity were determined.     

A Study of 5920 Strains of Psychrotrophic Bacteria Isolated from Chickens

The psychrotrotrophic flora of chicken carcases in 3 different industrial processing plants has been studied. The micro-organisms, 5920 strains, were placed in 4 categories (arranged in decreasing order of the number of isolates): (1), Pseudomonas and Achromobacter (Acineto-bacter) ; (2) Flavobacterium, Corynebacterium and yeasts; (3), Aeromonas and Entero-bacteriaceae; (4), lactic acid bacteria, Micrococcaceae and Bacillus . Different types of strains belonging to these groups are described and the importance of each is discussed with reference to the observations of other authors. The proteolytic action of 1717 of the 5920 isolates was studied.     

Extracellular protein A from a methicillin-resistant strain of Staphylococcus aureus

Protein A has been isolated from a methicillin-resistant strain of Staphylococcus aureus, A676, which produces only extracellular protein A. The material is homogeneous after a one-step separation and has a molecular weight of 41000. Its physicochemical and immunochemical properties have been studied.     

A Comparative Study of Staphylococcus aureus Strains Isolated from Bovine Subclinical Mastitis During 1952–1956 and 1992

Fiftytwo strains of S. aureus isolated from cases of bovine subclinical mastitis in 52 different dairy herds in Denmark, in the periods 1952 to 1956 and 1992, were compared with regard to their phage- and EcoRI ribo-types. Furthermore, susceptibility to penicillin and production of fibrinolysin were used as additional phenotypic markers. Fortynine strains (94%) could be separated into 12 phage types. Ribotyping assigned the 52 strains to 21 different types. Both methods showed that 57% of the 1950’s strains and between 38–45% of the 1992 strains belonged to 3 dominating types. The remaining strains were placed by ribotyping in 8 types occurring among the 1952–1956 strains and 10 types occurring among the 1992 strains. In 87% of the strains the results of the 2 typing methods were in accordance. However, 7 strains gave different results by the 2 methods including 2 strains with major differences. Penicillin resistance only occurred in a single genotype from the 1950’s compared to 6 different genotypes among the 1992 strains.     

雪貂葡萄球菌的分离与鉴定

目的对雪貂分离菌株进行鉴定。方法通过菌落、菌体形态、生化特征等细菌学检测方法和药敏实验对所分离菌株进行鉴定。结果根据生物学特性和药敏实验确定所分离菌株为金黄色葡萄球菌和中间葡萄球菌。结论分离菌株为金黄色葡萄球菌和中间葡萄球菌。     

Protein A Mutants of Staphylococcus aureus

Staphylococcus aureus Cowan I was exposed to nitrosoguanidine or ethyl-methanesulfonate, and survivors were screened on nutrient agar plates containing rabbit anti-protein A serum for loss of protein A production. More than half of all protein A-deficient mutants also lacked nuclease, coagulase, alpha hemolysin, fibrinolysin, mannitol utilization, and the phage-type pattern. Mutants with a spectrum of these properties were also isolated. Induced or spontaneous reversions of the mutants were observed. The properties of the protein A-deficient mutants suggest that synthesis or release (or both) of a number of extracellular products of S. aureus is controlled by a common regulatory mechanism.     

Extracellular Fibrinogen-binding Protein (Efb) from Staphylococcus aureus Inhibits the Formation of Platelet-Leukocyte Complexes

Extracellular fibrinogen-binding protein (Efb) from Staphylococcus aureus inhibits platelet activation, although its mechanism of action has not been established. In this study, we discovered that the N-terminal region of Efb (Efb-N) promotes platelet binding of fibrinogen and that Efb-N binding to platelets proceeds via two independent mechanisms: fibrinogen-mediated and fibrinogen-independent. By proteomic analysis of Efb-interacting proteins within platelets and confirmation by pulldown assays followed by immunoblotting, we identified P-selectin and multimerin-1 as novel Efb interaction partners. The interaction of both P-selectin and multimerin-1 with Efb is independent of fibrinogen. We focused on Efb interaction with P-selectin. Excess of P-selectin extracellular domain significantly impaired Efb binding by activated platelets, suggesting that P-selectin is the main receptor for Efb on the surface of activated platelets. Efb-N interaction with P-selectin inhibited P-selectin binding to its physiological ligand, P-selectin glycoprotein ligand-1 (PSGL-1), both in cell lysates and in cell-free assays. Because of the importance of P-selectin-PSGL-1 binding in the interaction between platelets and leukocytes, we tested human whole blood and found that Efb abolishes the formation of platelet-monocyte and platelet-granulocyte complexes. In summary, we present evidence that in addition to its documented antithrombotic activity, Efb can play an immunoregulatory role via inhibition of P-selectin-PSGL-1-dependent formation of platelet-leukocyte complexes.     

Comparison of Purified Alpha-Toxins from Various Strains of Staphylococcus aureus

Alpha-toxin from five strains of Staphylococcus aureus, including Wood 46, was purified by isoelectric focusing. The alpha-toxins obtained from different strains were similar. The isoelectric point of the purified toxins was 8.65 +/- 0.15. Sharp concentration peaks were not always obtained. In the ultracentrifuge the alpha-toxins migrated usually as three peaks which could be dissociated with propionic acid to yield one peak. A single line of identity was obtained in immunoelectrophoresis when a heterologous antiserum was reacted with the five purified toxins. It was concluded that the widespread use of the Wood 46 strain for the production of alpha-toxin is justified.     

Staphylococcus aureus Manganese Transport Protein C (MntC) Is an Extracellular Matrix- and Plasminogen-Binding Protein

Infections caused by Staphylococcus aureus – particularly nosocomial infections - represent a great concern. Usually, the early stage of pathogenesis consists on asymptomatic nasopharynx colonization, which could result in dissemination to other mucosal niches or invasion of sterile sites, such as blood. This pathogenic route depends on scavenging of nutrients as well as binding to and disrupting extracellular matrix (ECM). Manganese transport protein C (MntC), a conserved manganese-binding protein, takes part in this infectious scenario as an ion-scavenging factor and surprisingly as an ECM and coagulation cascade binding protein, as revealed in this work. This study showed a marked ability of MntC to bind to several ECM and coagulation cascade components, including laminin, collagen type IV, cellular and plasma fibronectin, plasminogen and fibrinogen by ELISA. The MntC binding to plasminogen appears to be related to the presence of surface-exposed lysines, since previous incubation with an analogue of lysine residue, ε-aminocaproic acid, or increasing ionic strength affected the interaction between MntC and plasminogen. MntC-bound plasminogen was converted to active plasmin in the presence of urokinase plasminogen activator (uPA). The newly released plasmin, in turn, acted in the cleavage of the α and β chains of fibrinogen. In conclusion, we describe a novel function for MntC that may help staphylococcal mucosal colonization and establishment of invasive disease, through the interaction with ECM and coagulation cascade host proteins. These data suggest that this potential virulence factor could be an adequate candidate to compose an anti-staphylococcal human vaccine formulation.     

Detecting the Enterotoxigenicity of Staphylococcus aureus Strains

An optimal sensitivity plate method for examining large number of staphylococcal strains for production of the known enterotoxins (A-E) is presented. Small volumes of relatively concentrated enterotoxin are produced by the semi-solid agar, cellophane-over-agar, or sac culture techniques. Detection of the enterotoxin in the supernatant fluid is accomplished with the optimal sensitivity plate method. In this method small plastic petri dishes (50 mm) were used for a modified Ouchterlony of high sensitivity.     

Apparent Spontaneous Induction of Staphylococcus aureus Isolated from Clinical Sources

Apparent spontaneous induction in staphylococcal strains from two clinical specimens was described. One of the two phages associated with these strains was found useful in typing otherwise untypable strains.     

Molecular Epidemiology of Methicillin-Resistant Staphylococcus aureus Isolated from Australian Veterinarians

This work investigated the molecular epidemiology and antimicrobial resistance of methicillin-resistant Staphylococcus aureus (MRSA) isolated from veterinarians in Australia in 2009. The collection (n = 44) was subjected to extensive molecular typing (MLST, spa, SCCmec, dru, PFGE, virulence and antimicrobial resistance genotyping) and antimicrobial resistance phenotyping by disk diffusion. MRSA was isolated from Australian veterinarians representing various occupational emphases. The isolate collection was dominated by MRSA strains belonging to clonal complex (CC) 8 and multilocus sequence type (ST) 22. CC8 MRSA (ST8-IV [2B], spa t064; and ST612-IV [2B], spa variable,) were strongly associated with equine practice veterinarians (OR = 17.5, 95% CI = 3.3–92.5, P < 0.001) and were often resistant to gentamicin and rifampicin. ST22-IV [2B], spa variable, were strongly associated with companion animal practice veterinarians (OR = 52.5, 95% CI = 5.2–532.7, P < 0.001) and were resistant to ciprofloxacin. A single pig practice veterinarian carried ST398-V [5C2], spa t1451. Equine practice and companion animal practice veterinarians frequently carried multiresistant-CC8 and ST22 MRSA, respectively, whereas only a single swine specialist carried MRSA ST398. The presence of these strains in veterinarians may be associated with specific antimicrobial administration practices in each animal species.     

Genetic Variation among Staphylococcus aureus Strains from Norwegian Bulk Milk

Strains of Staphylococcus aureus obtained from bovine (n = 117) and caprine (n = 114) bulk milk were characterized and compared with S. aureus strains from raw-milk products (n = 27), bovine mastitis specimens (n = 9), and human blood cultures (n = 39). All isolates were typed by pulsed-field gel electrophoresis (PFGE). In addition, subsets of isolates were characterized using multilocus sequence typing (MLST), multiplex PCR (m-PCR) for genes encoding nine of the staphylococcal enterotoxins (SE), and the cloverleaf method for penicillin resistance. A variety of genotypes were observed, and greater genetic diversity was found among bovine than caprine bulk milk isolates. Certain genotypes, with a wide geographic distribution, were common to bovine and caprine bulk milk and may represent ruminant-specialized S. aureus. Isolates with genotypes indistinguishable from those of strains from ruminant mastitis were frequently found in bulk milk, and strains with genotypes indistinguishable from those from bulk milk were observed in raw-milk products. This indicates that S. aureus from infected udders may contaminate bulk milk and, subsequently, raw-milk products. Human blood culture isolates were diverse and differed from isolates from other sources. Genotyping by PFGE, MLST, and m-PCR for SE genes largely corresponded. In general, isolates with indistinguishable PFGE banding patterns had the same SE gene profile and isolates with identical SE gene profiles were placed together in PFGE clusters. Phylogenetic analyses agreed with the division of MLST sequence types into clonal complexes, and isolates within the same clonal complex had the same SE gene profile. Furthermore, isolates within PFGE clusters generally belonged to the same clonal complex.