Antimicrobial susceptibility and invasive ability of <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> isolates from mastitis from dairy backyard systems

Fifteen (15) backyard farms were investigated to determine the antimicrobial susceptibility and invasion ability of S. aureus isolates from cows with subclinical mastitis in México. A total of 106 cows were sampled and 31 S. aureus isolates were recovered. S. aureus isolates were resistant to penicillin class antibiotics and susceptible to gentamicin and cetyltrimethylammonium bromide. STA9 and STA13 isolates were resistant to erythromycin (MIC > 25 microg/ml) and lincomycin (STA13, MIC > 25 microg/ml; STA9, MIC > 100 microg/ml). STA9 isolate harbors the erm(B) and msr(A) genes, whereas STA13 isolate harbors the erm(C) gene. STA9 and STA13 isolates contains the lnu(A) gene. Only 5 isolates (STA11, STA13, STA14, STA15 and STA21) were able to internalize in bovine mammary epithelial cells. These results indicate that S. aureus isolates from dairy backyard farms showed differences in the antimicrobial susceptibility patterns and invasion ability in bovine mammary epithelial cells. This kind of evaluations should be performed in different dairy regions, since resistance patterns and isolate diversity vary on a per-region basis.     

<Emphasis Type="Italic">In Vitro</Emphasis> activity of telithromycin and quinupristin/dalfopristin against methicillin-resistant coagulase-negative staphylococci with defined resistance genotypes

We determined the activities of new antibiotics telithromycin (ketolide) and quinupristin/dalfopristin (streptogramins) against 88 macrolide and/or lincosamide resistant coagulase-negative staphylococci (CoNS) isolates with defined resistance gene status. Telithromycin susceptibility was determined only in erythromycin-sensitive isolates (15) indicating the same mechanisms of resistance. In contrast, all erythromycin-resistant isolates (73) were either constitutively resistant to telithromycin (13 isolates with constitutive erm genes) or demonstrated telithromycin D-shaped zone (60 isolates with inducible msr(A) and/or erm). However, the level of inducible resistance conferred by msr(A) (35 isolates) was borderline even after induction by erythromycin. No quinupristin/dalfopristin resistant isolate was observed if tested by disk-diffusion method (DDM) but 18 isolates were intermediate (MIC = 1-3 mg/L) and two isolates resistant (MIC = 8 mg/L) if tested by E-test. All these isolates were resistant to streptogramin A and harbored vga(A) gene (1 isolate) or vga(A)LC gene (19 isolates). MICs for quinupristin/dalfopristin were higher for isolates with combination of streptogramin A resistance and constitutive MLSB resistance (MIC = 3-8 mg/L in 4 isolates) than for streptogramin A-resistant isolates susceptible to streptogramin B (MIC = 0.5-2 mg/L in 16 isolates). In addition to S. haemolyticus, vga(A)LC was newly identified in S. epidermidis and S. warnerii indicating its widespread occurrence in CoNS. Misidentification of low-level resistant isolates by DDM may contribute to dissemination of streptogramin A resistance.     

Staphylococcus aureus nosocomial infections: the role of a rapid and low-cost characterization for the establishment of a surveillance system

Continuous surveillance on resistance patterns and characterization of Staphylococcus aureus represent simple and low-cost techniques to understand and evaluate the effectiveness of infection control and antimicrobial prescribing measures. In this study we analyzed the antibiotic susceptibility and trends for S. aureus strains collected from bacteraemia cases in a five year period. Between 2004 and 2008 we noted a progressive decrease in the number of S. aureus isolates compared to all pathogens from clinical specimens and S. aureus bloodstream infections (BSI) reflected a similar trend. In particular we analyzed 185 isolates from blood cultures: 89 isolates were MSSA and 96 isolates were MRSA. Molecular SCCmec typing of these strains showed an absolute prevalence of types I and II, whereas five spa types from 96 isolates were obtained. Resistance pattern analysis allowed us to place MRSA strains into 12 antibiotypes and the major antibiotype was resistant to penicillin, gentamicin, erythromycin, clindamycin and ciprofloxacin. The predominant antibiotype among the MSSA isolates was resistant only to penicillin. In addition, 19.1% of MSSA are susceptible to all antibiotics tested. We also found a close association between antibiotyping 1 and genotyping t002/SCCmecI of MRSA strains, suggesting a nosocomial scenario dominated by a few particular clones.     

金黄色葡萄球菌所致肺部感染的耐药特点及其Panton—Valentine杀白细胞素基因的携带状况分析

目的分析金黄色葡萄球菌所致肺部感染的耐药性特点及其Panton—Valentine杀白细胞素基因的携带状况。方法回顾性调查了温州医学院第一附属医院2005年1月至2006年1月医院感染的金黄色葡萄球菌所致肺部感染患者132例,对其体外药敏试验进行分析;并利用多重PCR检测其PVL基因,应用多位点基因序列分型（multilocus sequence typing,MLST）技术对PVL基因阳性的菌株进行序列分型。耐甲氧西林金黄色葡萄球菌（methicillin-resistant Staphylococcus aureus,MRSA）的SCCmec基因分型采用多重聚合酶链反应。结果致肺部感染的132株金黄色葡萄球菌的耐药现象较为严重,仅对万古霉素、呋喃妥因及复方新诺明等药物的敏感率较高;其中经多重PCR筛选出10株携带PVL基因的金葡菌,全部为MRSA菌株,3株为ST239-SCCⅢ,2株为ST398-SCCmecⅢ,2株为ST398-SCCmecⅣ,ST25-SCCmecⅢ、ST59-SCCmecⅠ和ST88-SCCmecⅢ各1株。结论肺部感染的金黄色葡萄球菌对多种抗生素耐药,呈多重耐药性;其携带PVL基因占一定比例。     

Cephapirin: In Vitro Antibacterial Spectrum

Cephapirin, a new semisynthetic cephalosporin derivative, was found to have an antibacterial spectrum similar to that of cephalothin. Staphylococcus aureus was inhibited by cephapirin concentrations of 0.09 to 12.5 mug/ml. S. epidermidis, S. viridans, S. pyogenes, and Diplococcus pneumonia isolates were inhibited by less than 1 mug/ml. The Enterococcus required a concentration of 25 mug of antibiotic per ml for inhibition. Approximately 65% of Escherichia coli, and all Klebsiella, indole-negative Proteus, and Salmonella strains tested were inhibited by the drug. Serratia, Pseudomonas, indole-positive Proteus, and Erwinia strains were highly resistant. Inoculum size was not an important factor in determining the level of sensitivity of S. aureus to cephapirin. The antibiotic does not appear to be significantly bound to serum protein. In vitro development of resistance to the drug was demonstrated with two isolates of S. aureus.     

Antibiotic resistance dynamics and isolation rate of staphylococci and enterococci from patients of reconstructive surgery units

The dynamics of isolation of staphylococci and enterococci from clinical material of patients and their antibiotic susceptibility within a 5-year period (2005-2009) was analysed. 5990 isolates were tested: 1250 isolates of Staphylococcus aureus, 3268 isolates of S. epidermidis, 1005 isolates of Enterococcus faecalis and 467 isolates of E. faecium. Grampositive infections were shown to be prevailing within the last 2-3 years, the nosocomial epidermal staphylococci more and more replacing S. aureus (the ratio of S. epidermidis and S. aureus in 2009 was 3.3). The isolation rate of E. faecalis significantly increased (by 3.5 times) and the ratio of E. faecalis and E. faecium in 2009 was 4.3. The microflora composition with respect to the isolation source was analysed and its clinical significance was estimated. The study of the antibiotic susceptibility showed that oxacillin had its own specific niche, while antibiotics active against resistant grampositive cocci, such as rifampicin, fusidic acid, fluoroquinolones (moxifloxacin), cefoxitin, as well as amoxicillin/clavulane in infections due to E. faecalis, might be considered as the drugs of choice. In the treatment of nosocomial infections, when the etiological role of MRSA or VRE is suspected or confirmed, the complex therapy should obligatory include the most active antibiotics (vancomycin or linezolid among them).     

Effects of various types of Triton X on the susceptibilities of methicillin-resistant staphylococci to oxacillin

We examined the effect of six types of the nonionic detergent Triton X on the susceptibilities of methicillin-resistant staphylococci to oxacillin. We used five methicillin-resistant Staphylococcus aureus isolates and 17 methicillin-resistant coagulase-negative staphylococci isolates. All strains of S. aureus, S. epidermidis and S. sciuri had enhanced susceptibility to oxacillin following exposure to the types of Triton X having 7–13 polymerized ethylene oxides. These strains were altered from homogeneously resistant to heterogeneously resistant by Triton X-100. Those types of Triton X that affected the resistance level also promoted the release of lipoteichoic acid. These results and those of previous studies suggest that Triton X might act on factors other than the mecA or femA products.     

Characterization of Staphylococcus aureus strains isolated from raw milk of bovine subclinical mastitis in Tehran and Mashhad

Staphylococcus aureus is considered one of the most important food borne pathogens. A total of 111 isolates of S. aureus were cultured from raw milk samples during January 2009 to June 2009 from Tehran and Mashhad. The coagulase gene polymorphism and the prevalence of classical enterotoxin genes of S. aureus strains were determined by PCR-RFLP (restriction fragment length polymorphism) and Multiplex-PCR. Disk diffusion method was used to determine the susceptibility of isolates to antimicrobial agents as instructed by Clinical and Laboratory Standards Institute. Sixty-seven % of the isolates harboured one or more enterotoxin genes. The most prevalent gene was sec, found in 59 % of the isolates. Approximately 8% of the isolates were positive for sea, seb and sed genes. Only one isolate had see gene. The rate of coexistence of enterotoxin genes was 14%. All S. aureus isolates were susceptible to ciprofloxacin, gentamicin, imipenem, minocycline, oxacillin and vancomycin. They were resistant to ampicillin (64%), penicillin (56%), clindamycin (22%), tetracycline (22%), doxycycline (19%), teicoplanin (13%), rifampin (2%) and trimethoprim-sulfamethoxazole (2%). On the basis of coagulase gene analysis of 111 S. aureus isolates, the PCR products of 56 isolates were digested with Alu I that produced three distinct patterns. These data indicate the high prevalence of enterotoxigenic S. aureus in raw bovine milk in Tehran and Mashhad, and highlight the importance of proper quality control of dairy products for public health.     

长沙地区临床分离金黄色葡萄球菌的耐药监测

目的了解长沙地区临床分离金黄色葡萄球菌（以下简称金葡菌）对常用抗菌药物的耐药现状,探讨金黄色葡萄球菌对甲氧西林的耐药水平。方法收集长沙地区11家医院2009年11月至2010年11月临床分离的非重复金葡菌279株,应用Vitek-2全自动微生物分析系统进行鉴定,K-B法检测金葡菌对24种药物的敏感性,产色头孢菌素试验检测β-内酰胺酶以及D试验检测诱导型克林霉素耐药。应用头孢西丁和苯唑西林纸片扩散法筛查耐甲氧西林的金葡菌（MRSA）,琼脂稀释法检测头孢西丁和苯唑西林的最低抑菌浓度（MIC）。结果在被检测的24种药物中,敏感率〉50%的药物为9种,未发现对万古霉素、替考拉宁和利奈唑胺耐药菌株;耐药率〉50%的抗菌药物有11种,其中以青霉素和氨苄西林的耐药率最高（均为97.1%）。MRSA的分离率达54.5%,且对常用的16种抗菌药物的耐药率均显著高于甲氧西林敏感金黄色葡萄球菌（MSSA）。279株金葡菌中,β-内酰胺酶阳性250株（89.6%）;红霉素耐药而克林霉素敏感或中介的30株中,D试验阳性22株（73.3%）。苯唑西林（OXA）和头孢西丁（FOX）MIC范围分别为0.125～〉256μg/mL和2～〉256μg/mL,苯唑西林的MIC50和MIC90分别为128μg/mL和256μg/mL,头孢西丁的MIC50和MIC90分别为64μg/mL和256μg/mL。结论长沙地区临床分离金葡菌对常用抗菌药物呈多重耐药;MRSA不仅分离率高,而且对甲氧西林呈高水平耐药。     

Methicillin (Oxacillin)-resistant Staphylococcus aureus strains isolated from major food animals and their potential transmission to humans

From May 2001 to April 2003, various types of specimens from cattle, pigs, and chickens were collected and examined for the presence of methicillin (oxacillin)-resistant Staphylococcus aureus (MRSA). S. aureus was isolated and positively identified by using Gram staining, colony morphology, tests for coagulase and urease activities, and an API Staph Ident system. Among 1,913 specimens collected from the animals, 421 contained S. aureus; of these, 28 contained S. aureus resistant to concentrations of oxacillin higher than 2 micro g/ml. Isolates from 15 of the 28 specimens were positive by PCR for the mecA gene. Of the 15 mecA-positive MRSA isolates, 12 were from dairy cows and 3 were from chickens. Antimicrobial susceptibility tests of mecA-positive MRSA strains were performed by the disk diffusion method. All isolates were resistant to members of the penicillin family, such as ampicillin, oxacillin, and penicillin. All isolates were also susceptible to amikacin, vancomycin, and trimethoprim-sulfamethoxazole. To determine molecular epidemiological relatedness of these 15 animal MRSA isolates to isolates from humans, random amplified polymorphic DNA (RAPD) patterns were generated by arbitrarily primed PCR. The RAPD patterns of six of the isolates from animals were identical to the patterns of certain isolates from humans. The antibiotypes of the six animal isolates revealed types similar to those of the human isolates. These data suggested that the genomes of the six animal MRSA isolates were very closely related to those of some human MRSA isolates and were a possible source of human infections caused by consuming contaminated food products made from these animals.     

Etiology of severe hospital infections in intensive care units and antibiotic resistance of pathogens]

The leading pathogens of severe infections in intensive care units were the following: respiratory tract infections--bacteria of the famility of Enterobacteriaceae (33.8%), Pseudomonas spp. (24.9%), Acinetobacter spp. (18.1%), Staphylococcus aureus (16.0%), blood flow infections--coagulase negative staphylococci (33.6%), S. aureus (26.1%), Enterobacteriaceae (17.6%), wound infections--Enterobacteriaceae (35.7%), coagulase negative staphyloccocci (17.8%), Pseudomonas spp. (14.3%). As for various species of Enterobacteriaceae, susceptibility was preserved in 91-100% of the isolates to meropenem, in 72-100% to cefoperazone/sulbactam, in 51-65% to cefepime, in 72-86% to amikacin, and in less than 50% to cephalosporins and fluoroquinolones. As for P.aeruginosa, 28% of the isolates was resistant to all the antibacterials, except polymyxin. The highest susceptibility to cefoperazone/sulbactam and meropenem was revealed in the isolates of Acinetobacter baumannii. Oxacillin resistance was detected in 64.9% of the S.aureus isolates. The oxacillin resistance as a rule was associated with resistance to macrolides, aminoglycosides and fluoroquinolones. As for coagulase negative staphylococci, oxacillin resistance was stated in 75.6% of the isolates. All the isolates of the Staphylococcus spp. preserved their susceptibility to vancomycin and linezolid.     

Properties of the strains<Emphasis Type="Italic">Enterococcus haemoperoxidus</Emphasis> and<Emphasis Type="Italic">E. moraviensis</Emphasis>, new species among enterococci

Antibiotic susceptibility or resistance, urease activity, detection of the structural genes for bacteriocin production, bacteriocin activity as well as sensitivity of the isolates to enterocins (Ent) A and M were determined in 23 isolates of new species Enterococcus haemoperoxidus and E. moraviensis. The majority of the strains were antibiotic sensitive and exhibited low urease activity (< 10 nkat/mL). The most frequently detected genes for Ent were entA and entP. However, only the strain 466 of E. haemoperoxidus produced an antibacterial substance with inhibitory activity against 21 G+ indicators. It was partially purified reaching an activity of up to 12 800 AU/mL. This bacteriocin active strain also possessed the genes for EntA and EntP. The other strains did not inhibit the indicator strains. The substance produced by the 466 strain was active even after a 5-months storage at +4 and -20 degrees C. This substance has proteolytic and hydrophilic character, pH optimum of bacteriocin production by this strain being between 4 and 7. While E. moraviensis strains showed sensitivity to EntA (produced by E. faecium EK13) and to EntM (produced by E. faecium AL41), E. haemoperoxidus strains were sensitive to EntA (except strain 382) but less sensitive to the treatment by EntM.     

Nasal carriage of methicillin resistant Staphylococcus aureus and their antibiotic susceptibility patterns in children attending day-care centers

Nasal colonization with community acquired methicillin resistant Staphylococcus aureus (CA-MRSA) is being increasingly reported, especially in places where people are in close contact and in reduced hygiene, such as day-care centers. In this study we investigated the frequency of MRSA colonization and their antibiotic susceptibility patterns in 1-6 years old children of day-care centers in Hamadan, West of Iran.Five hundred nasal swabs were collected from children of 27 day-care centers that had no risk factors for colonization by S. aureus. The specimens were cultured for isolation of S. aureus by standard methods. Antimicrobial susceptibility testing was performed according to the Clinical and Laboratory Standards Institute (CLSI) guidelines. For evaluation of the frequency of erythromycin induced clindamycin resistance, disk approximation test (D-test) was applied.Totally, 148 (29.6%) children were colonized by S. aureus. Out of 260 male, 94 (36.2%) and of 240 female, 54 (22.5%) cases were nasal carriers of S. aureus (P value = 0.001). Six (4.1%) of the 148 S. aureus isolated from children were MRSA strains. None of MRSA and methicillin susceptible S. aureus (MSSA) was resistant to vancomycin and clindamycin. Three of the 6 strains of MRSA and 7 (4.9%) of the 142 MSSA strains were resistant to erythromycin, and D-test was positive in all of them.We conclude that the rate of colonization by S. aureus is high in children attending day-care centers but colonization with MRSA is not common in our areas. Clindamycin or trimethoprim-sulfamethoxazol could be used in mild to moderataly severe diseases caused by CA-MRSA. However, if the CA-MRSA isolates are erythromycin resistant, D-test should be carried out for detection of inducible clindamycin resistance.     

Detection of methicillin- and aminoglycoside-resistant genes and simultaneous identification of S. aureus using triplex real-time PCR Taqman assay

In this study we describe a triplex real-time PCR assay that enables the identification of S. aureus and detection of two important antibiotic resistant genes simultaneously using real-time PCR technology in a single assay. In this triplex real-time PCR assay, the mecA (methicillin resistant), femA (species specific S. aureus) and aacA-aphD (aminoglycoside resistant) genes were detected in a single test using dual-labeled Taqman probes. The assay gives simultaneous information for the identification of S. aureus and detection of methicillin and aminoglycoside resistance in staphylococcal isolates. 152 clinical isolates were subjected to this triplex real-time PCR assay. The results of the triplex real-time PCR assay correlated with the results of the phenotypic antibiotic susceptibility testing. The results obtained from triplex real-time PCR assay shows that the primer and probe sets were specific for the identification of S. aureus and were able to detect methicillin- and aminoglycoside-resistant genes. The entire assay can be performed within 3 h which is a very rapid method that can give simultaneous information for the identification of S. aureus and antibiotic resistance pattern of a staphylococcal isolate. The application of this rapid method in microbiology laboratories would be a valuable tool for the rapid identification of the S. aureus isolates and determination of their antibiotic resistance pattern with regards to methicillin and aminoglycosides.     

Characterization of Staphylococcus aureus Strains Associated with Food Poisoning in Shenzhen, China

To characterize isolates of Staphylococcus aureus that were associated with staphylococcal food poisoning between 2006 and 2009 in Shenzhen, Southern China, a total of 52 Staphylococcus aureus isolates from 11 outbreaks were analyzed by using multilocus sequence typing (MLST), spa typing, and pulsed-field gel electrophoresis (PFGE). PCR analysis was used to analyze the staphylococcal enterotoxin (SE) genes sea to sei, and antimicrobial susceptibility testing was also performed. ST6 was the most dominant sequence type (ST), constituting 63.5% (34/52) of all of the isolates in 7 outbreaks. The next most common ST was ST943, which constituted 23.1% (12/52) of the isolates that were collected from 3 outbreaks. t701, t091, and t2360 were the most predominant spa types, constituting 67.3% (35/52) of the isolates that were collected from 11 outbreaks. Three PFGE types, (types A, B, and C) were the most frequently observed types, constituting 84.6% (44/52) of all of the isolates. The enterotoxin gene that we detected most frequently was sea (45/52; 86.5%). Four SE gene profiles were observed, including sea (n = 45), sec-seh (n = 3), seb (n = 2), and seg-sei (n = 2). With respect to antibiotic resistance, penicillin resistance was the most common (96.2%; 50/52), followed by resistance to tetracycline (28.8%; 15/52). Approximately 30.8% (16/52) of the isolates were resistant to at least two antibiotics, and 7.7% (4/52) of the isolates were resistant to three or more drugs. The two predominant S. aureus lineages, (i) PFGE types A and B with ST6 and (ii) PFGE type C with ST943, were identified in the outbreaks.     

濒危植物浮叶慈姑遗传多样性的RAPD分析

采用RAPD分子标记技术,对内蒙古二卡河(EK)和黑龙江库都尔(KDE)的浮叶慈姑居群进行了遗传多样性分析。 从60个随机引物中筛选出14个引物对2个居群共48个样品进行扩增,得到93条清晰的条带,其中70条具有多态性,即物 种水平上的多态位点百分率(PPB)为75.27%。POPGENE分析结果表明:二卡河居群的PPB为66.67%,库都尔居群的PPB 为72.04%。两个居群间基因分化系数(Gst)为0.0628,即遗传变异有很大一部分是来自居群内(93.72%)。结合相关的分析 表明生态学因素可能是浮叶慈姑濒危的主要原因。     

猪源致病性金黄色葡萄球菌的分离鉴定及其耐药性分析

目的鉴定引起猪渗出性皮炎的病原,并分析猪源致病性金黄色葡萄球菌的耐药性,为临床用药提供依据。方法采集患渗出性皮炎的仔猪标本进行细菌分离培养,联合应用形态学检查、生理生化试验和PCR方法鉴定分离菌株,并进行致病性和药物敏感性试验。结果先后从病猪标本中分离鉴定获得PSA1、PSA2、PSA3和PSA4四株金黄色葡萄球菌,其中PSA1和PSA3分离株的致病性较强。药敏试验结果显示PSA1、PSA2和PSA3分离株为MRSA菌株,PSA4分离株为MSSA菌株。MRSA菌株对14种抗菌药物均呈现不同程度的耐药,尤其是对青霉素、链霉素、四环素、强力霉素、环丙沙星和氧氟沙星等6种抗菌药物的耐药率达100%。所有分离株对万古霉素与替考拉宁均敏感。结论合肥地区猪渗出性表皮炎的病原为金黄色葡萄球菌。猪源致病性金黄色葡萄球菌合肥分离株具有多重耐药性,治疗猪渗出性皮炎应建立在体外药敏试验的基础上,有针对性选择抗菌药物。     

Effect of subinhibitory levels of selected antibiotics on susceptibility of Staphylococcus aureus strains to phagocytosis and killing by rabbit granulocytes

The aim of the study was to assess the of chosen antibiotics in subinhibitory concentrations (sub-MIC) on the sensitive of Staphylococcus aureus cells to phagocytosis and killing by rabbit granulocytes. The following antibiotics were used: cloxacillin, cefadroxil, cefuroxim, cefotaxim, gentamicin, netilmicin, lincomicin, doxycycline and riphamicin. A total of 144 S. aureus strains with varied sensitivity to these antibiotics were selected for the study. The experiment used granulocytes isolated from rabbit blood and S. aureus strains incubated for 18 h in TSB broth containing antibiotics in the concentrations of 0.1 MIC, 0.2 MIC and 0.5 MIC, and in the antibiotics-free medium. Phagocytosis was assessed by the method of differential staining with acridine orange and crystal violet, allowing simultaneous determination of phagocytised and killed S. aureus cell counts. The findings revealed that the culture of S. aureus in the presence of all the antibiotics used in subinhibitory concentrations increased significantly the susceptibility of most S. aureus strains to phagocytosis and killing by granulocytes. The above effect usually occurred in the concentrations of 0.1 MIC (54.2%), more seldom in 0.2 MIC (13%) and 0.5 MIC (15% of strains). Each group of S. aureus contained some which showed no change in susceptibility following culture with the chemotherapeutic agents in subinhibitory concentrations (26.3%). Insensitive strains to the subinhibitory effects were equally common among susceptible (27%), intermediate (23%) and resistant (26%) strains of S. aureus to the antibiotics used. No statistically significant reduction was noted in phagocytosis or killing by rabbit granulocytes. No correlation was observed between the susceptibility to the subinhibitory effects of the antibiotics involved and their biochemical mechanisms.     

The structure and antibiotic sensitivity of the causative agents of community-acquired infectious diseases of bacterial origin in children]

Four hundred and forty pediatric patients at the age of 7 days to 15 years with various infections admitted to the Hospital within a month were examined. The biological material was inoculated to blood agar on the first days of the patient admittance to the Hospital and after the growth the organisms were isolated and identified. Antibiotic susceptibility of the isolates was assayed with the disk diffusion method. 479 strains in all were tested. The most frequent cases requiring hospitalization and antibiotic therapy were those of respiratory tract infections (54.09 per cent), urinary tract infections (26.36 per cent), cutaneous and subcutaneous fat diseases, gastrointestinal diseases and others (about 25 per cent of the cases in all). The main pathogens were Streptococcus viridans, S.aureus and S.epidermidis, as well as Enterobacteriaceae (chiefly E.coli) whose frequencies were practically equal (in 25-35 per cent of the cases). The Pneumococcus isolates amounted to 6.3 per cent. Nonfermenting bacteria (Pseudomonas aeruginosa and Acinetobacter) and some representatives of Enterobacteriaceae (Citrobacter, Serratia, Morganella) were isolated from 7 per cent of the patients. The frequency of Klebsiella and Enterobacter was about 11 per cent. The main pathogens were tested for their susceptibility to amoxycillin/clavulanic acid, ampicillin, oxacillin and gentamicin. The least active antibiotic was ampicillin. 88.8 per cent of the E.coli isolates and 100 per cent of the Klebsiella, P.mirabilis, Morganella, Citrobacter, Enterobacter and Serratia isolates were resistant to it. 53.2 per cent of the Streptococcus isolates including 64.5 per cent of the Pneumococcus isolates were as well resistant to ampicillin. 59.5 per cent of the Streptococcus isolates (mainly S.viridans and Enterococcus) was susceptible to oxacillin, 22.2 per cent of them being moderately susceptible. 62.5 per cent of the Pneumococcus isolates and 78.1 per cent of the Staphylococcus isolates were also susceptible to oxacillin. The highest susceptibility of the isolates was that to amoxycillin/clavulanic acid, i.e. 90.1 per cent of the strains, 79.9 per cent of them being highly susceptible. All the isolates of Citrobacter, Serratia and Morganella and some isolates of P.aeruginosa, Acinetobacter, Enterobacter, Klebsiella and E.coli were resistant to amoxycillin/clavulanic acid. As for the latter 5 organisms their susceptibility to amoxycillin/clavulanic acid was comparable with that to gentamicin. The susceptibility of the Streptococcus and Staphylococcus isolates to amoxycillin/clavulanic acid was significantly much higher than that to oxacillin, gentamicin and ampicillin: 93 per cent of the Streptococcus isolates (62.7 per cent of the Pneumococcus isolates) and 90.7 per cent of the Staphylococcus isolates.     

Alterations of cell wall structure and metabolism accompany reduced susceptibility to vancomycin in an isogenic series of clinical isolates of Staphylococcus aureus

A series of isogenic methicillin-resistant Staphylococcus aureus isolates recovered from a bacteremic patient were shown to acquire gradually increasing levels of resistance to vancomycin during chemotherapy with the drug (K. Sieradzki, T. Leski, L. Borio, J. Dick, and A. Tomasz, J. Clin. Microbiol. 41:1687-1693, 2003). We compared properties of the earliest (parental) vancomycin-susceptible isolate, JH1 (MIC, 1 microg/ml), to two late (progeny) isolates, JH9 and JH14 (vancomycin MIC, 8 microg/ml). The resistant isolates produced abnormally thick cell walls and poorly separated cells when grown in antibiotic-free medium. Chemical analysis of the resistant isolates showed decreased cross-linkage of the peptidoglycan and drastically reduced levels of PBP4 as determined by the fluorographic assay. Resistant isolates showed reduced rates of cell wall turnover and autolysis. In vitro hydrolysis of resistant cell walls by autolytic extracts prepared from either susceptible or resistant strains was also slow, and this abnormality could be traced to a quantitative (or qualitative) change in the wall teichoic acid component of resistant isolates. Some change in the structure and/or metabolism of teichoic acids appears to be an important component of the mechanism of decreased susceptibility to vancomycin in S. aureus.