Antagonistic action of Pseudomonas aeruginosa against Staphylococci, coliform bacilli and various types of Proteus

Antagonistic activity of 2 fresh isolates and 3 collection strains of Pseudomonas aeruginosa against 177 microbial strains was determined with the method of late antagonism. Among the microbial strains there were 56 staphylococcal strains isolated from patents and carriers. 38 nontypable colon bacilli isolated from healthy persons, 59 enteropathogenic colon bacilli of various serogroups, 12 strains of Proteus and 12 colon bacilli, carriers of multiple drug resistance factors (R factors). All the cultures were sensitive to the antagonistic action of 5 or at least 3 strains of Pseudomonas used in the study. The most active antagonists were the fresh isolates of Pseudomonas as compared to the collection strains. Among the staphylococci S. aureus proved to be the most resistant to the antagonistic action of Pseudomonas as compared to S. epidermidis, the same as the strains isolated from carriers as compared to the strains isolated from patients. As for the enteric bacilli the most resistant were the strains of Proteus. Acquiring of transmissive R factors by the colon bacilli markedly increased their sensitivity to the antagonistic action of Pseudomonas.     

Antagonistic action of corynebacteria and bacilli of a skin ecotype on staphylococci

582 strain of corynebacteria and 235 cultures of bacilli isolated from the skin surface of mammary glands of 120 nursing women were studied for their antagonistic action on staphylococci. It is established that bacilli, especially Bacillus subtilis, B. firmus and B. alvei exert more pronounced inhibitory effect on staphylococci. Representatives of the family Corynebacteriaceae possess a weak antagonistic action both on coagulase-positive and on coagulase-negative species. The ability of coryneform bacteria to retain the colonization resistance is supposed to be due to other factors. Antagonistic properties of B. subtilis are shown expedient to be used for elimination of hospital strains of Staphylococcus aureus from the skin of mammary glands.     

Diagnostic value of persistent characteristics of staphylococci during their carriage

Comparative assessment of information value of significant persistent characteristics of staphylococci in state of carriage was made, and the usefulness of these characteristics for creating diagnostic models was defined. One hundred and fifty strains of Staphylococcus aureus and S. epidermidis isolated from healthy persons with subsequent differentiation of carriage types were used in the study. It has been established that strains isolated from resident carriers more often have more markedly expressed factors of persistence. Ranking the factors of persistence of staphylococci according to their information value during carriage allowed to identifythe leading sign among them--anti-carnosine activity. Using factor analysis, obtained signs were united in the single complex defining bacterial carriage, which allows to build a mathematical model permitting to diagnose staphylococcal carriage with 95% probability.     

Direct detection of human Staphylococcus aureus carriage in the nose using the Lightcycler Staphylococcus kit

The Lightcycler Staphylococcus kit is a diagnostic tool for direct real-time detection of Staphylococcus aureus in clinical materials. We show here that detection of S. aureus nasal carriage using this test is hampered by competition of DNA from coagulase-negative staphylococci. However the test is well suited for species identification after culture and the identification of high-load S. aureus carriers.     

金黄色葡萄球菌拮抗菌株的筛选鉴定及其抗菌物质分析

【背景】植物内生菌的次生代谢产物是新型天然活性物质的重要来源。【目的】从芍药内生细菌中筛选对金黄色葡萄球菌有抑菌活性的菌株和次生代谢产物。【方法】采用平板对峙法筛选拮抗菌株,根据形态学特征和分子生物学的方法鉴定菌株,PCR扩增检测合成脂肽类物质的功能基因;运用牛津杯法依次测定内生细菌发酵液和脂肽类粗提物的抑菌活性,利用Sephadex LH-20凝胶层析分离脂肽类物质,利用基质辅助激光解吸电离飞行时间质谱分析具有抑菌作用的分离组分。【结果】共筛选出13株对金黄色葡萄球菌具有不同程度抑制作用的内生菌株,其中菌株SY11的抑菌作用最为显著,其发酵液和脂肽类粗提物均具有较强的抑制作用。结合形态学鉴定以及16S r RNA基因序列分析,鉴定其为解淀粉芽孢杆菌(Bacillusamyloliquefaciens)。PCR扩增检测表明菌株SY11含有3个合成脂肽类物质的功能基因fenA、ituD和srfkn,推测该菌株可能具有合成脂肽类物质的能力。根据具有抑菌活性分离组分的质谱分析结果,推测其有效物质的主要成分为Bacillomycin D。【结论】解淀粉芽孢杆菌SY11对金黄色葡萄球菌有良好抑制效果,其脂肽类粗提物也具有较强的体外抑菌活性。本研究为芍药内生细菌的开发应用奠定了基础。     

The role of anti-lactoferrin activity of bacteria in their persistence

The role of anti-lactoferrin activity (ALfA) of bacteria in the persistence phenomenon, i.e. long survival of a causative agent in a host body, was studied in experimental staphylococcal infection. In experimental animals infected with isogenic clones of Staphylococcus aureus with different level of ALfA, longer terms were noted in bacterial recovery of staphylococci with this ability. Analysis on the occurrence and level of ALfA in clinical isolates of bacteria, isolated from different forms of infectious process, confirmed the significance of this sign in the persistence of a causative agent. High values of penetrance and expression of ALfA in bacteria were revealed in strains of microorganisms, isolated from chronic inflammatory diseases and from bacterial carriers.     

Antibiotic sensitivity of the microflora isolated in ENT diseases]

Sensitivity of beta-hemolytic streptococci of group A, Streptococcus viridans, Staphylococcus aureus, epidermal staphylococci, pneumococci, Proteus and Ps. aeruginosa isolated in 1975-1978 from patients with tonsillitis, otitis, sinusitis and other otorhinolaryngological diseases was studied with respect to 19 antibiotics. Data on comparison of the antibiotic sensitivity of the microflora isolated from the patients with otorhinolaryngological diseases in 1964-1974 and 1975-1978 are presented. It was shown that beta-hemolytic streptococci were highly sensitive to all the antibiotics tested except tetracycline. Among Streptococcus viridans the strains resistant to many antibiotics were more frequent than among beta-hemolytic streptococci. Most of the Staphylococcus aureus strains were sensitive to gentamycin, cephaloridin, oxacillin and resistant to the other antibiotics. The epidermal staphylococci were characterized by approximately the same antibiotic sensitivity as Staphylococcus aureus. Resistance of the predominating majority of the Pneumococcus strains to tetracycline was noted. Proteus and Ps. aeruginosa were resistant to all the antibiotics except aminoglycosides. The microflora isolated from the cases with otorhinolaryngological diseases in 1975-1978 were mainly characterized by lower antibiotic sensitivity than that isolated from the cases with the same diseases in 1964-1974. It is possible to suppose that the microorganisms isolated from the patients with otorhinolaringological diseases had no significant differences with respect to their antibiotic sensitivity from those isolated from the patients with other pathological processes.     

Simultaneous PCR detection of ica cluster and methicillin and mupirocin resistance genes in catheter-isolated Staphylococcus.

Recent data show that more than 50% of catheter-associated bloodstream infections are caused by staphylococci. Staphylococcal infections produced by intercellular-adhesion cluster (ica) carriers can be even more problematic due to the presence of methicillin and mupirocin resistance genes. In the present study, a multiplex PCR protocol that allows the simultaneous identification of staphylococci and detection of both the ica and methicillin and/or mupirocin resistance genes was developed. Furthermore, the method allows differential detection of the ica locus from Staphylococcus aureus and Staphylococcus epidermidis.     

Detection of the capsule of staphylococci isolated in urological diseases]

In studying the cultures isolated from urological patients the seed material was found to yield a high proportion (40%) of coagulase-negative, white staphylococci. Capsule-forming organisms constituted 58.8% of the isolated Staphylococcus aureus. As the presence of capsules is indicative of pathogenicity of staphylococci, in urological diseases the presence of capsulary strains in urine should be taken into consideration.     

Characteristics of staphylococci isolated from man, poultry and some other animals

Of 281 strains of staphylococci isolated from man and animals 36 (12.8%) were coagulase-positive and 245 (87.2%) were coagulase-negative. Staphylococcus aureus and Staph. intermedius were the commonest coagulase-positive staphylococci isolated from the hosts examined. Of the 20 strains that remained unclassifiable, 14 were isolated from sheep and goats.     

Desoxyribonuclease and lecithinase activity in antibiotic-resistant and -sensitive staphylococci]

Staphylococcus aureus, a laboratory strain 209-P and strain I isolated freshly from infected wounds, as well as lincomycin hydrochloride, ampicillin, oxacillin and methicillin manufactured in the USSR and cephaloridin manufactured by "PLIVA" in Yugoslavia were used. Various activity levels of desoxyribonuclease and lecitinase of the staphylococci depending on sensitivity or resistance of the test-microbe to the antibiotics were shown. The activity of the above microbial enzymes characterizing the pathogenic properties decreased with development of the antibiotic resistance, sometimes to complete inactivation of the enzymes synthesized by the staphylococci. In spite of closeness of their modes of action the semisynthetic penicillins had a differentiating effect on the above enzymes.     

Action of egg lysozyme on representatives of the family Micrococcaceae. The action on staphylococci

The results of the study on the effect of various concentrations of lysozyme on staphylococci of three species, i.e. Staphylococcus aureus, Staphylococcus epidermidis and Staphylococcus saprophyticus are presented. The study was performed with the use of the turbidimetric method. It was found that all the staphylococci had a common tendency for changing the optic density (OD) of the suspensions under the effect of lysozyme which was different from that in Micrococcus luteus. However, the dynamics of decreasing the OD in Staph. saprophyticus was characterized by some properties which provided a supposition that the structures of the cell walls in Staph. aureus and Staph. epidermidis were close, while the cell walls in Staph. saprophyticus had some unknown characteristics.     

Characteristics of staphylococci isolated from man, poultry and some other animals

Of 281 strains of staphylococci isolated from man and animals 36 (12–8%) were coagulase-positive and 245 (87–2%) were coagulase-negative. Staphylococcus aureus and Staph. intermedius were the commonest coagulase-positive staphylococci isolated from the hosts examined. Of the 20 strains that remained unclassifiable, 14 were isolated from sheep and goats.     

Identification of staphylococci from bovine udders: evaluation of the API 20GP system

The API 20GP system (Analytab Products, Plainview, NY) correctly identified 56.1% of staphylococci isolated from the bovine mammary gland. The system identified 90.2% of Staphylococcus aureus strains, but failed to recognize strains of Staphylcoccus hyicus and others. Poor performance was attributed to the limited number of veterinary strains contained in the profile index data base. The API 20GP was determined to be an unacceptable method for identification of staphylococci isolated from the bovine mammary gland.     

医院工作人员鼻腔葡萄球菌带菌状况年度推移的调查报告

本文报道1989年从202名医院工作人员鼻腔分离葡萄球菌的带菌状况和药敏性检测结果。并将此结果同作者等于1985年对200名同类人员检测的结果相比较,作了年度推移的调查分析发现1989年度的总带菌率(76.6％)比1985年度(84.5％)略有下降,但其中的金黄色葡萄球菌带菌率却从1985年的7.5％上升为10.4％。从1989年和1985两个年度的调查中,均发现临床科室人员金葡菌带菌率高于其它辅助科室人员,说明医务人员带菌率与接触病人成正相关关系。分离菌株对12种抗菌药物的耐药性检测结果显示,1989年分离菌株对其中9种的敏感性下降,并且从耐药谱显示出对5种抗菌药物耐药的多重耐药菌株明显增多。另从金葡菌的耐药情况,也看出1989年的耐药率高于1985年菌株。还在1989年分离的金葡菌中出现25％的耐甲氧西林菌株。     

Presence of enterotoxin C and toxic shock syndrome toxin--1 (TSST-1) genes in population of Staphylococcus aureus phage type 187

The aim of this study was to examine whether Staphylococcus aureus of phage type 187 possess the genes of enterotoxins and toxic shock syndrom toxin. Sixteen phage type 187 strains were isolated from the hospital patients (12) and the carriers (4) in twelve medical centres in Poland during 1991 and 2005. Biotyping, phage typing, antibiotic susceptibility, detection of the genes of enterotoxins (sea--sed) and toxic shock syndrome toxin (tst) was tested. The results of this study showed that all staphylococci of phage type 187 belonged to the human biotype (A) and appeared to be sensitive to all of the tested antibiotics, including methicillin (MSSA). Almost all of them (93.8%) had the enterotoxin C gene and TSST-1 gene. This fact allows to consider them the strains of potentially high virulence.     

905株葡萄球菌的种群分布及耐药性分析

目的了解某医院2010年临床分离葡萄球菌的种群分布和耐药性。方法对该院2010年临床分离的905株葡萄球菌的种群、耐药性做回顾性分析。结果905株葡萄球菌包含11个种,其中金黄色葡萄球菌（SAU）455株,占50．28％;凝固酶阴性葡萄球菌（CNS）450株,占49．72％;临床分离葡萄球菌对抗生素耐药率可因标本来源、培养环境、种群结构、产酶与否等因素差异有统计学意义。除利奈唑胺、万古霉素和呋喃妥因敏感及四环素耐药率差异无统计学意义以外,耐甲氧西林葡萄球菌（MRS）高于甲氧西林敏感葡萄球菌（MSS）,x2值在4．30—451．0,P〈0．01或0．05,差异有统计学意义;β-内酰胺酶阳性菌高于β-内酰胺酶阴性菌,凝固酶阴性的葡萄球菌与凝固酶阳性菌对利福平、氨苄西林／舒巴坦、青霉素G、四环素、左氧氟沙星、庆大霉素的耐药率差异存在统计学意义,P〈0．05或P〈0．01;尿菌高于其他标本分离菌。各种葡萄球菌在各临床科室及标本中的分布也不尽相同。结论葡萄球菌耐药性可由多方面因素引起,临床实验室必须加强葡萄球菌分布和耐药性监测,为临床提供各种葡萄球菌诊治的依据。     

Use of resistant mutants to study the interaction of triton X-100 with Staphylococcus aureus.

Staphylococcus aureus mutants resistant to the nonionic detergent Triton X-100, isolated from the wild-type strain H and the autolysin-deficient strain RUS3, could grow and divide in broth containing 5% (vol/vol) Triton X-100, while growth of the parental strains was markedly inhibited above the critical micellar concentration (0.02%) of the detergent. Growth-inhibitory concentrations of Triton X-100 killed wild-type cells without demonstrable cellular lysis. Triton X-100 stimulated autolysin activity of S. aureus cells under nongrowing conditions, and this lytic response was markedly reduced in energy-poisoned cells. In contrast, the detergent had no effect on the activity of autolysins in cell-free systems, and growth in the presence of Triton X-100 did not alter either the cellular autolysin activity or the susceptibility of cell walls to exogenous lytic enzymes. Treatment with either Triton X-100 or penicillin G in the growth medium stimulated release of predominantly acylated intracellular lipoteichoic acid and sensitized staphylococci to Triton X-100-induced autolysis. There was no significant difference in the cell wall and membrane compositions or Triton X-100 binding between the parental strains and the resistant mutants. The resistant mutant TXR1, derived from S. aureus H, had a higher level of L-alpha-glycerophosphate dehydrogenase activity, and its oxygen uptake was more resistant to inhibition by a submicellar concentration (0.008%) of Triton X-100. Growth in the presence of subinhibitory concentrations of Triton X-100 rendered S. aureus H cells phenotypically resistant to the detergent and greatly stimulated the level of oxygen uptake. Membranes isolated from such cells exhibited enhanced activity of the respiratory enzymes succinic dehydrogenase and L-alpha-glycerophosphate dehydrogenase.     

Resistance of different strains of pathogenic staphylococcus to unfavorable environmental factors]

A study was made of the resistance to drying the UV-irradiation, the action of furacillin and chloramine displayed by 60 stains of S. aureus differing by origin (hospital and extrahospital), by the source of discharge (the upper respiratory tracts of carriers and the discharge of the purulent-inflammatory foci of surgical patients), relation to the antibiotics (polyresistant and sensitive) and phage-group reference. It was found that the resistance of staphylococci to the unfavourable factors was not always associated with the listed signs of the strains. In respect to drying a marked resistance was expressed by the hospital strains in comparison with the extrahospital ones, polyresistant in comparison with the sensitive ones, staphylococci of III and I+III phage groups in comparison with the strains of other bacteriophage groups. Strains of the III phage group proved to be the most resistant to the UV-irradiation. Strains isolated from carriers were more resistant to furacillin than staphylococci isolated from the purulent-inflammatory foci. Strains of the III phage group and nontyping had analogous advantages over the cultures of other phage groups.     

Lysis of staphylococcal mastitis pathogens by bacteriophage phi11 endolysin

The Staphylococcus aureus bacteriophage phi11 endolysin has two peptidoglycan hydrolase domains (endopeptidase and amidase) and an SH3b cell wall-binding domain. In turbidity reduction assays, the purified protein can lyse untreated staphylococcal mastitis pathogens, Staphylococcus aureus and coagulase-negative staphylococci (Staphylococcus chronogenes, Staphylococcus epidermidis, Staphylococcus hyicus, Staphylococcus simulans, Staphylococcus warneri and Staphylococcus xylosus), making it a strong candidate protein antimicrobial. This lytic activity is maintained at the pH (6.7), and the "free" calcium concentration (3 mM) of milk. Truncated endolysin-derived proteins containing only the endopeptidase domain also lyse staphylococci in the absence of the SH3b-binding domain.