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PubMed1964

DEMONSTRATION OF INDUCED SYNERGISTIC HEMOLYSIS BY "NONHEMOLYTIC" STAPHYLOCOCCUS SPECIES.

Smith, Doyle C. (Kansas State University, Manhattan), V. D. Foltz, and T. H. Lord. Demonstration of induced synergistic hemolysis by "non-hemolytic" Staphylococcus species. J. Bacteriol. 87:188-195. 1964.-The synergistic hemolysis of "normally nonhemolytic" staphylococci on blood-agar incubated in, or adjacent to, a secondary zone of certain hemolytic staphylococci is described. The synergism apparently results from the combining of two factors, Z, produced by hemolytic staphylococci that elaborate a secondary zone, and T, produced by "nonhemolytic" staphylococci. The production of the two factors is substantiated by (i) the absence of clear hemolysis around nonhemolytic colonies and in the secondary zone of hemolytic colonies, and by (ii) evidence of clear hemolysis when the secondary zone reaches to within a few millimeters, and finally surrounds, the nonhemolytic colonies. It was possible to show on a blood-agar plate containing a mixture of both hemolytic and nonhemolytic cultures that all colonies exhibited a zone of clear hemolysis. When 400 colonies were selected from such a blood plate, 280 were nonhemolytic on subsequent studies. The remaining 120 colonies were hemolytic. Thus, there is danger of confusing non-hemolytic Staphylococcus colonies, showing induced hemolysis, with truly hemolytic colonies. If a similar situation occurred on a diagnostic blood plate, it could very likely result in failure to identify a possibly pathogenic staphylococcus.

Agar↗

PubMed2004

New quadriplex PCR assay for detection of methicillin and mupirocin resistance and simultaneous discrimination of Staphylococcus aureus from coagulase-negative staphylococci.

Major challenges in diagnostic molecular microbiology are to develop a simple assay to distinguish Staphylococcus aureus from the less virulent but clinically important coagulase-negative staphylococci (CoNS) and to simultaneously determine their antibiotic resistance profiles. Multiplex PCR assays have been developed for the detection of methicillin- and mupirocin-resistant S. aureus and CoNS but not for the simultaneous discrimination of S. aureus from CoNS. We designed a new set of Staphylococcus genus-specific primers and developed a novel quadriplex PCR assay targeting the 16S rRNA (Staphylococcus genus specific), nuc (S. aureus species specific), mecA (a determinant of methicillin resistance), and mupA (a determinant of mupirocin resistance) genes to identify most staphylococci, to discriminate S. aureus from CoNS and other bacteria, and to simultaneously detect methicillin and mupirocin resistance. Validation of the assay with 96 ATCC control strains and 323 previously characterized clinical isolates, including methicillin- and mupirocin-sensitive and -resistant S. aureus and CoNS isolates and other bacteria, demonstrated 100% sensitivity, specificity, and accuracy. This assay represents a simple, rapid, accurate, and reliable approach for the detection of methicillin- and mupirocin-resistant staphylococci and offers the hope of preventing their widespread dissemination through early and reliable detection.

Anti-Bacterial Agents↗

PubMed2004

Simultaneous detection of Staphylococcus aureus and coagulase-negative staphylococci in positive blood cultures by real-time PCR with two fluorescence resonance energy transfer probe sets.

A real-time PCR assay that uses two fluorescence resonance energy transfer probe sets and targets the tuf gene of staphylococci is described here. One probe set detects the Staphylococcus genus, whereas the other probe set is specific for Staphylococcus aureus. One hundred thirty-eight cultured isolates, which contained 41 isolates of staphylococci representing at least nine species, and 100 positive blood cultures that contained gram-positive cocci in clusters were tested. This assay was 100% sensitive and 100% specific for the detection of the Staphylococcus genus and of S. aureus.

Blood↗

PubMed1991

Comparison of the pathogenicity of three species of coagulase-negative Staphylococcus in a mouse model with and without a foreign body.

Staphylococcus schleiferi, Staphylococcus lugdunensis, and Staphylococcus epidermidis produce a high incidence of abscesses in a mouse model with an implanted foreign body. We investigated the significance of the foreign body in this process. Fourteen strains of S. schleiferi, S. epidermidis, and S. lugdunensis were tested in our model. A preadhered foreign body was implanted into one mouse group, followed by injection of a test strain. Another group received injection without implant. Abscesses were assessed at 7 days; foreign bodies and infected tissues were cultured. The percent of samples that developed abscesses or were culture positive was compared for each strain. Nearly all mice infected with S. schleiferi developed abscesses and were culture positive. The foreign body made no difference in abscess formation for three of four S. schleiferi but increased the incidence of both organism recovery and abscess for three of five S. epidermidis. The foreign body enhanced abscess formation for four of five S. lugdunensis, with all five strains yielding significantly more culture recovery. Although the pathogenicity of nine strains was increased by the foreign body, five strains yielded high abscess and culture recovery rates that were not enhanced by its presence.

Abscess↗

PubMed2005

Pathogenesis of Staphylococcus in the rabbit anterior chamber.

PURPOSE: To investigate the host defense against Staphylococcus in the rabbit anterior chamber. METHODS: The bactericidal activity of rabbit aqueous humor was investigated in vitro. Rabbit anterior chambers were injected with viable Staphylococcus aureus or Staphylococcus epidermidis (1,000 or 500,000 colony-forming units [CFU]), killed bacteria, culture supernatants of either organism, or purified S. aureus alpha-toxin. CFU as well as phospholipase (PLA(2)) and myeloperoxidase (MPO) activities of aqueous humor were determined up to 25 hours postinfection (PI). RESULTS: The number of viable S. aureus or S. epidermidis was significantly reduced when incubated with aqueous humor for 30 minutes (P </= 0.0001). Rabbits challenged with either S. aureus or S. epidermidis demonstrated a significant reduction in CFU in aqueous humor by 1 hour PI (P </= 0.0044). Eyes infected with either S. aureus or S. epidermidis demonstrated a significant increase in MPO activity beginning at 1 hour PI (P </= 0.0455), but only S. aureus caused an increase in PLA(2) activity at 20 and 25 hours PI (P </= 0.0002). No significant increases in PLA(2) activity were observed after injection of killed bacteria into the aqueous humor at any time point; however, injection of S. aureus supernatant or alpha-toxin into the anterior chamber significantly increased PLA(2) activity (P </= 0.0210). Injection of alpha-toxin also resulted in significant increases in MPO activity beginning at 10 hours after injection (P </= 0.001). CONCLUSIONS: This study demonstrates that aqueous humor has a potent host defense capability and that S. aureus, but not S. epidermidis, triggers a PLA(2) response in the rabbit anterior chamber that appears to be due to alpha-toxin.

Animals↗

PubMed2024

Genomic characterization of methicillin-resistant Staphylococcus aureus isolated from patients attending regional referral hospitals in Tanzania.

BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) colonization increases the risk of subsequent infection by MRSA strain complex interlinking between hospital and community-acquired MRSA which increases the chance of drug resistance and severity of the disease. OBJECTIVE: Genomic characterization of Staphylococcus aures strains isolated from patients attending regional referral hospitals in Tanzania. METHODOLOGY: A laboratory-based cross-sectional study using short read-based sequencing technology, (Nextseq550,Illumina, Inc. San diego, California, USA). The samples used were collected from patients attending selected regional referral hospitals in Tanzania under the SeqAfrica project. Sequences were analyzed using tools available in the center for genomic and epidemiology server, and visualization of the phylogenetic tree was performed in ITOL 6.0. SPSS 28.0 was used for statistical analysis. RESULTS: Among 103 sequences of S. aureus, 48.5% (50/103) carry the mecA gene for MRSA. High proportions of MRSA were observed among participants aged between 18 and 34 years (52.4%), in females (54.3%), and among outpatients (60.5%). The majority of observed MRSA carried plasmids rep5a (92.0%), rep16 (90.0%), rep7c (90.0%), rep15 (82.0%), rep19 (80.0%) and rep10 (72.0%). Among all plasmids observed rep5a, rep16, rep20, and repUS70 carried the blaZ gene, rep10 carried the erm(C) gene and rep7a carried the tet(K) gene. MLST and phylogeny analysis reveal high diversity among MRSA. Six different clones were observed circulating at selected regional hospitals and MRSA with ST8 was dominant. CONCLUSION: The study reveals a significant presence of MRSA in Staphylococcus aureus strains from Tanzanian regional hospitals, with nearly half carrying the mecA gene. MRSA is notably prevalent among young adults, females, and outpatients, showing high genetic diversity and dominance of ST8. Various plasmids carrying resistance genes indicate a complex resistance profile, highlighting the need for targeted interventions to manage MRSA infections in Tanzania.

Humans↗

PubMed1975

[Factors influencing transduction of genetic determinants of penicillinase activity and pathogenicity in staphylococcus aureua. II. Antiphage activity of acridine derivatives].

Acridine dyes examined earlier (acrichine, acridine orange, proflavine and rivanol) and newly-synthesized preparations (acridines No. No. 37--40) were studied in the capacity of nonspecific agents influencing the lytic cycle in development of staphylococcus phages. Acrichine and acridine No. 37 failed to prevent lysis of the indicator staphylococcus cultures (strains 16/160 and 8325) by bacteriophages; proflavine, rivanol, acridines No. No. 39--40 produced a marked inhibitory effect; acridine orange and acridine No. 38 inhibited the staphylococcus lysis completely. Some preparations could be used to investigate the transduction phenomenon.

Acridines↗

PubMed1988

Inactivation of lincosaminide antibiotics in Staphylococcus. Identification of lincosaminide O-nucleotidyltransferases and comparison of the corresponding resistance genes.

Resistance to lincomycin by inactivation has been detected in numerous clinical isolates of Staphylococcus; in crude extracts of Staphylococcus haemolyticus BM4610 and Staphylococcus aureus BM4611, inactivation of lincomycin and clindamycin requires the presence of a nucleoside 5'-triphosphate (ATP, GTP, CTP, or UTP) as nucleotidyl donor and Mg2+ as cofactor. The biochemical mechanism of lincosaminide inactivation was elucidated by determination of the structure of inactivated lincomycin and clindamycin by physicochemical techniques, including UV absorption spectrophotometry, 31P and 1H nuclear magnetic resonance, and periodate oxidation. In the two strains, inactivation of lincomycin gave rise to lincomycin 3-(5'-adenylate), whereas clindamycin was inactivated through its conversion to clindamycin 4-(5'-adenylate). The gene linA' encoding the 3-lincomycin, 4-clindamycin O-nucleotidyltransferase in S. aureus BM4611 has been sequenced and displays 93% homology with the gene linA encoding the 3-lincomycin, 4-clindamycin O-nucleotidyltransferase found in S. haemolyticus BM4610. The two enzymes are 161 amino acids long and differ by 14 amino acid substitutions.

Amino Acid Sequence↗

PubMed1986

Frequency and antimicrobial susceptibility of Staphylococcus species isolated from canine pyodermas.

Specimens obtained from pyogenic skin lesions of 210 dogs were culturally examined for staphylococci. A total of 215 isolates of staphylococci were biotyped, using the biochemical tests contained in a commercial staphylococcal identification system. Of 201 coagulase-positive isolates, 197 were identified as Staphylococcus intermedius, 3 as S aureus, and 1 as S hyicus. Of 14 coagulase-negative isolates, 5 were identified as S epidermidis, 5 as S xylosus, 3 as S simulans, and 1 as S hominis. Antimicrobial susceptibility tests were done on all staphylococcal isolates, using the standard disk-diffusion method. Staphylococcus intermedius isolates were susceptible to cephalothin, methicillin, and gentamicin. Resistance to ampicillin, penicillin G, and tetracycline was frequent. Antibiotic resistance was not associated with the depth of skin infection. Resistance to ampicillin, penicillin, tetracycline, and trimethoprim-sulfamethoxazole was not associated with previous antibiotic use. Increased resistance to chloramphenicol, clindamycin, and erythromycin was associated with previous antibiotic therapy. Antimicrobial susceptibilities of the other Staphylococcus species isolated are reported, but the small numbers of these species precluded making meaningful comparison with S intermedius.

Animals↗

PubMed1983

Biotypes, serotypes and phage types of caprine strains of Staphylococcus aureus.

Nine of 11 caprine strains of Staphylococcus aureus examined belong to biotype C. They were typable serologically using antisera developed for typing human strains of Staphylococcus aureus. H2 was found to be the dominant thermostable antigen whilst 1 and 0 were the most common thermolabile antigens. Some strains possessed protein A and poly A beta (beta-glucosaminyl ribitol teichoic acid). Phage typing of the strains was achieved with an international set of phages for typing human strains of Staphylococcus aureus and they belong to phage groups II, and the mixed group.

Animals↗

PubMed1994

Detection of methicillin-resistant staphylococcus aureus in human saliva and on denture surfaces.

The prevalence of Staphylococcus species and methicillin-resistant Staphylococcus aureus (MRSA) in human saliva and on denture surfaces was examined with selective media. A total of 166 saliva samples from dental students and individuals using dentures and 39 swab samples from denture surfaces were subjected in this study. The detected percentage of MRSA from saliva samples was 2.3%. Five out of 39 swab samples of denture surface contained MRSA or methicillin-resistant Staphylococcus epidermidis (MRSE). The isolated MRSA and MRSE were resistant to beta-lactam antibiotics.

Adult↗

PubMed1984

Staphylococcus epidermidis septicemia in children with leukemia and lymphoma.

Staphylococcus epidermidis is emerging as a cause of morbidity and mortality in immunocompromised patients. From January 1980 through June 1982, there were 150 episodes of septicemia in 92 children with leukemia and lymphoma at Memorial Sloan-Kettering Cancer Center, New York. Staphylococcus epidermidis was the fourth most common organism isolated, responsible for 12.7% of all septicemic episodes. Only nine of 53 isolates were sensitive to methicillin; all were sensitive to vancomycin. Staphylococcus epidermidis septicemia was associated with immunosuppressive chemotherapy (94.7%); broad-spectrum antibiotics (79.0%); catheters and drains (73.7%); neutropenia (63.2%); skin or soft-tissue infections (42.1%); prior septicemia (42.1%); concurrent polymicrobial septicemia (21.1%); and prolonged hospitalization (mean, 39 days). Of 19 patients, two died. Increased awareness of the pathogenic potential of S epidermidis in children with hematologic malignancies and prompt alteration of therapy to an effective antimicrobial agent, in most cases vancomycin hydrochloride, is required when the organism is isolated in patients known to be at risk with clinical evidence of septicemia.

Adolescent↗

PubMed1989

Neonatal Staphylococcus epidermidis meningitis with unremarkable CSF examination results.

We identified 10 infants (mean +/- SD birth weight, 1000 +/- 500 g; gestation, 29 +/- 3 weeks; postnatal age, 24 +/- 19 days) who had Staphylococcus epidermidis meningitis despite unremarkable cerebrospinal fluid (CSF) blood cell counts and glucose and protein levels. Staphylococcus epidermidis meningitis was diagnosed if all the following criteria were satisfied: (1) a CSF culture positive for S epidermidis within 48 hours, (2) a blood culture positive for S epidermidis with antibiotic sensitivities identical to those of the CSF isolate, and (3) clinical symptomatology. Lumbar puncture yielded white blood cell counts lower than 10 x 10(6)/L in 8 infants. Two subjects had CSF white blood cell counts of 11 x 10(6)/L and 14 x 10(6)/L. Cerebrospinal fluid glucose (2.8 +/- 0.9 mmol/L) and protein (1.15 +/- 0.32 g/L) concentrations were also unremarkable. Infants were treated with parenteral antibiotics for 19 +/- 5 days. There was no mortality or short-term morbidity. Staphylococcus epidermidis is a recognized cause of nosocomial meningitis in low-birth-weight infants and frequently occurs without CSF abnormalities.

Cerebrospinal Fluid Proteins↗

PubMed1983

Wound healing accelerated by Staphylococcus aureus.

While comparing the effects on wound healing of a heated scalpel with those of the cold scalpel, we discovered that inoculation of rat skin incisions with a strain of Staphylococcus aureus dramatically accelerated the gain in wound strength. The accelerating effect was evident four days postoperatively, was maximal at seven to ten days, and was still present at 28 days. The accelerating effect was correlated with the number of S aureus organisms introduced into the wound, and was found in conventional rats and rats germ free up to the time of monocontamination with S aureus. There was no evidence of infection on gross examination; on histologic examination an occasional microabscess was seen in some rats. There may be both local and systemic mechanisms underlying the S aureus accelerating effect. Seven strains of S aureus with varying characteristics demonstrated the wound-healing accelerating effect. In sharp contrast, Staphylococcus epidermidis (three strains), Staphylococcus hominis (one strain), and Pseudomonas aeruginosa (two strains) did not show this effect. The increases in wound healing due to S aureus were substantially greater than reported previously for any nutritional supplement, drug, or other chemical or physical agent.

Animals↗

PubMed2005

Effects of controlled fibronectin surface orientation on subsequent Staphylococcus epidermidis adhesion.

Several bacterial species, including Staphylococcus aureus and Staphylococcus epidermidis (SE) are known to express cell receptors that bind specifically to surface immobilized or extracellular matrix ligands, such as the protein fibronectin (FN). Yet, few existing studies have examined the effect of protein surface orientation on bacterial adhesion. We report here a substratum modification protocol that allows for the specific orientation of FN molecules on a surface at known levels of surface coverage. Monoclonal antibodies (Mabs), specific to either the COOH-terminus or NH3-terminus of FN, are conjugated to biotin, then immobilized to streptavidin-coated glass substrata. Specific orientation of the bound FN molecules is verified using the same Mabs in an ELISA. Bacterial adhesion of Staphylococcus epidermidis (SE) to FN bound by either its C-terminus or its NH3-terminus was quantified in batch static adhesion assays. Results indicate an increase in SE adhesion to FN-coated surfaces when the FN is bound by its C-terminus (NH3-terminus free), indicating SE receptor-specific adhesion to the FN NH3-terminus. These studies demonstrate that antifibronectin monoclonal antibodies can be used to specifically bind and orient fibronectin on a surface. In addition, adhesion of SE to these model substrata can be controlled by the orientation of the protein.

Antibodies, Monoclonal↗

PubMed2006

Continentalic acid from Aralia continentalis shows activity against methicillin-resistant Staphylococcus aureus.

In a continuing search for compounds with antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA), a chloroform extract of roots of Aralia continentalis was found to contain continentalic acid (CA, C(20)H(30)O(2)), a diterpenic acid. This compound exhibited potent activity against standard methicillin-susceptible Staphylococcus aureus (MSSA) as well as clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA). It was determined that continentalic acid had minimum inhibitory concentrations (MICs) of approximately 8-16 microg/mL against S. aureus, including the MSSA and MRSA standard strains. Therefore, the results obtained in this study suggest that continentalic acid might have potential as an adjunct in the treatment of antibiotic-resistant bacteria.

Aralia↗

PubMed1998

Effect of lactoferrin on the phagocytic activity of polymorphonuclear leucocytes isolated from blood of patients with autoimmune diseases and Staphylococcus aureus allergy.

Phagocytic number (PN) and phagocytic index (PI) of neutrophils isolated from blood of patients with autoimmune diseases, allergy to Staphylococcus aureus and from blood of healthy individuals were examined. Our results concerning the influence of lactoferrin (Lf); (6.7 mg/l) on the PI of PMN showed that: 1) Lf enhances reliable PI of PMN at the 30-th minute starting the phagocytic reaction in patients with autoimmune disease in an active stage, in blood donors treated as healthy with the presence of autoantibodies, in patients with autoimmune diseases and proved autoantibodies against tissue, cell antigens and collagen, 2) Lf influences non-significantly PI of PMN in patients with autoimmune collagen diseases in remission, 3) Lf increases PI of PMN with 19% only in 58% from the assessed patients with Staphylococcus aureus, and 4) Lf decreases non-significantly PI of PMN in the healthy controls. Our studies on the effect of Lf on the phagocytic activity of PMN suggest that Lf has stronger effect on the PN compared to the PI: 1) Lf enhances with 86% the PN in patients with Staphylococcus aureus, 2) Lf increases PN of PMN in all of the assessed patients with autoimmune collagen diseases in active stage (mean with 72%), and 3) Lf increases PN of PMN in 4 from the 5 investigated healthy controls (mean with 22%). Our results show a "corrective" effect of Lf on the phagocytic functions in the investigated groups of patients. The possible mechanisms, by which Lf increases PN and PI of neutrophils, is discussed: 1) they may concern the antioxidative properties of Lf to block the iron ions in their catalytic inactive form or to take part as ferric-Lf in an oxidative-reduction processes on the plasma membrane and controlling transmembrane transport systems, 2) Lf decreases the negative surface charge and thus enhances the adherent ability of the PMN. Probably to this stimulated adherent ability dues the increased ingestion of bacteria in the presence of Lf, and 3) The "changed" membrane of PMN may have higher number receptors for Lf to bind more molecules of exogenous Lf. The increase of Lf binding which enhances the adherence and aggregation of neutrophils, facilitates the phagocytosis.

Adolescent↗

PubMed1993

Cloning and characterization of the scrA gene encoding the sucrose-specific Enzyme II of the phosphotransferase system from Staphylococcus xylosus.

By insertional mutagenesis with the staphylococcal transposon Tn551, mutants of Staphylococcus xylosus were isolated that were unable to utilize sucrose. One of these was found to be deficient in sucrose uptake. The genomic region containing this sucrose uptake gene of Staphylococcus xylosus (scrA) was cloned in Staphylococcus carnosus. The scrA gene was further localized to a 4.4 kb DNA fragment by complementation of the sucrose transport-deficient S. xylosus mutant. The DNA sequence analysis of the scrA region revealed three open reading frames, one of which encodes a protein of 480 amino acids (51.335 kDa) with significant similarity to sucrose-specific Enzymes II of phosphoenolpyruvate-dependent carbohydrate phosphotransferase systems (PTS). A protein with an apparent molecular weight of 50 kDa was obtained in Escherichia coli by expression of scrA with the bacteriophage T7 RNA polymerase promoter system. Transcriptional start sites of the scrA gene were localized by primer extension analysis to positions 46 and 49 nucleotides upstream of the scrA start codon. No additional sucrose utilization genes are encoded close to scrA on the S. xylosus chromosome.

Amino Acid Sequence↗