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Frequency and characteristics of selected enteropathogens in fecal and rectal specimens from childhood diarrhea in Trinidad, 1998-2000.

OBJECTIVE: To determine the prevalence and characteristics of selected enteric pathogens in diarrheic children in six counties of the island of Trinidad. METHODS: This cross-sectional study was conducted from April 1998 through March 2000. Fecal or rectal swab specimens from children (<12 years) were collected and then processed, using standard methods, to detect Salmonella spp., Shigella spp., enteropathogenic Escherichia coli (EPEC), Campylobacter spp., Yersinia spp., Cryptosporidium parvum, and parasite ova. The antibiograms of the enteropathogens were determined using the disk diffusion method. RESULTS: A total of 236 samples were processed; 86 samples originated directly from 17 heath centers in two counties (St. George East and St. George West), while 150 samples were obtained from the Trinidad Public Health Laboratory, having been submitted by private practitioners and personnel from six counties in Trinidad. Of the 236 samples, 33 (14.0%) were positive for Shigella, 4 (1.7%) for Salmonella, and 1 (0.4%) for EPEC. Two of the samples (0.8%) were positive for Campylobacter jejuni, while 1 sample (0.4%) was positive for hookworm ova. All the samples were negative for Cryptosporidium parvum and Yersinia spp. With the 86 samples collected directly from the health centers, in St. George East County the frequency of Shigella was 20.0% (12 of 60), compared with 26.9% (7 of 26) for samples from St. George West County, but the difference was not statistically significant (P>0.05 with the chi-square test). For the 150 samples from the six counties that had been submitted directly to the Trinidad Public Health Laboratory, 14 of them (9.3%) were positive for Shigella, a figure statistically significantly lower than that found with the samples sampled directly from the health centers (P<0.05 with the chi-square test). Sh. sonnei was the predominant serotype detected, accounting for 28 of the 33 Shigella isolates (84.8%) recovered from the 236 samples. Overall, the frequency of detection of enteropathogens had no seasonal pattern nor relationship to the county of origin. Of the 37 isolates of Salmonella and Shigella tested for antimicrobial sensitivity, all of them were sensitive to ciprofloxacin, gentamicin, and cefotaxime. In terms of resistance, 3 of the 37 isolates (8.1%) exhibited resistance to ampicillin, 1 (2.7%) to chloramphenicol, and 1 (2.7%) to sulfamethoxazole/ trimethoprim. CONCLUSION: Of the enteropathogens for which assays were done, Shigella sonnei was the most prevalent, and it has the highest probability of being an important etiological agent of childhood diarrhea in Trinidad.

Animals↗

Antimicrobial susceptibilities and plasmid contents of Neisseria gonorrhoeae isolates from commercial sex workers in Dhaka, Bangladesh: emergence of high-level resistance to ciprofloxacin.

Commercial sex workers (CSWs) serve as the most important reservoir of sexually transmitted diseases (STD), including gonorrhea. Periodic monitoring of the antimicrobial susceptibility profile of Neisseria gonorrhoeae in a high-risk population provides essential clues regarding the rapidly changing pattern of antimicrobial susceptibilities. A study concerning the prevalence of gonococcal infection among CSWs was conducted in Bangladesh. The isolates were examined with regards to their antimicrobial susceptibility to, and the MICs of, penicillin, tetracycline, ciprofloxacin, cefuroxime, ceftriaxone, and spectinomycin by disk diffusion and agar dilution methods. The total plasmid profile of the isolates was also analyzed. Of the 224 CSWs, 94 (42%) were culture positive for N. gonorrhoeae. There was a good correlation between the results of the disk diffusion and agar dilution methods. Some 66% of the isolates were resistant to penicillin, and 34% were moderately susceptible to penicillin. Among the resistant isolates, 23.4% were penicillinase-producing N. gonorrhoeae (PPNG). 60.6% of the isolates were resistant and 38.3% were moderately susceptible to tetracycline, 17.5% were tetracycline-resistant N. gonorrhoeae, 11.7% were resistant and 26.6% had reduced susceptibility to ciprofloxacin, 2.1% were resistant and 11.7% had reduced susceptibility to cefuroxime, and 1% were resistant to ceftriaxone. All PPNG isolates contained a 3.2-MDa African type of plasmid, and a 24.2-MDa conjugative plasmid was present in 34.1% of the isolates. Since quinolones such as ciprofloxacin are recommended as the first line of therapy for gonorrhea, the emergence of significant resistance to ciprofloxacin will limit the usefulness of this drug for treatment of gonorrhea in Bangladesh.

Anti-Infective Agents↗

Antimicrobial resistance in selected bacterial enteropathogens in north India.

BACKGROUND & OBJECTIVES: The resistance of enteropathogenic bacteria to commonly prescribed antibiotics is increasing both in developing as well as in developed countries. Resistance has emerged even to newer, more potent antimicrobial agents. The present study was therefore undertaken to report the current antibiotic resistance in common bacterial enteropathogens isolated in a tertiary care hospital in north India. METHODS: Faecal samples from 1802 patients were cultured for common bacterial enteropathogens and identified by standard methods. Antibiotic susceptibility was done by Stoke's disk diffusion method. The clinical and demographic profile of the patients was noted. RESULTS: Stool specimens from 119 (88 male, 31 female) patients yielded Shigella, Salmonella, Vibrio cholerae or Aeromonas. Fifty two per cent (62/119) of patients were children and 70 per cent were below the age of 5 yr. Twenty seven patients developed hospital acquired diarrhoea. Among all enteropathogens, Shigella spp. was the commonest followed by non-typhoidal Salmonella (27), V. cholerae O1 El tor serotype Ogawa (19), Aeromonas spp. (14), Salmonella Typhi and S. paratyphi A (2 isolates each). Resistance to antimicrobial agents was common among all pathogens. Among shigellae an overall resistance of 63.6, 58.1 and 16.3 per cent was observed for nalidixic acid, cotrimoxazole and furazolidone respectively. Seven isolates of Shigella were resistant to ciprofloxacin, (18.5%) of non-typhoidal salmonellae were resistant to ciprofloxacin. V. cholerae were generally susceptible to tetracycline (only 1 isolate out of 13 resistant) and other drugs except nalidixic acid (89.5% resistance) and cotrimoxazole (77.8% resistance). INTERPRETATION & CONCLUSION: Enteropathogens have developed high level resistance to first line agents used for empiric treatment of diarrhoea. Progressively increasing resistance to ciprofloxacin is a serious cause of concern.

Adult↗

The virulence factors and antibiotic sensitivities of Escherichia coli isolated from recurrent urinary tract infections.

OBJECTIVE: We determined the antibiotic sensitivities of uropathogenic Escherichia coli (UPEC) strains isolated from the urine of patients who have recurrent urinary tract infections (UTIs). METHODS: Our study was carried out between November 2000 and January 2002 at the Infectious Diseases Clinic, Istanbul Haydarpasa Numune Hospital, Istanbul, Turkey. We compared the virulence factors (fimbrial adhesion, hemolysin production, motility property) of 50 strains of Escherichia coli (E. coli) isolated from urine with the same properties of 25 strains of E. coli isolated from stool specimens of healthy individuals. In addition, we detected the virulence factors of UPEC strains using a microbiological and biochemical methods and by using disk diffusion method, we were able to investigate the sensitivity of the strains to the antimicrobials. RESULTS: We found the level of mannose-resistant (MR) fimbriae bearing in the UPEC strains to be significantly higher than that in the controls (odds ratio=10.27, p<0.001). The difference in mannose-resistant hemoagglutination (MRHA) and mannose sensitive hemoagglutination (MSHA) bearing levels in UPEC strains were rather high. This difference was regarded as significant in terms of showing the virulence of fimbriae bearing strains (odds ratio=29.03, p<0.001). CONCLUSION: Our study demonstrates that strains with MR fimbriae have a rather high virulence (p<0.001), and that a combination of MR+MS fimbriae increased that virulence (p<0.001). As MR strains have a greater adhesive property, the determination of MR fimbriae bearing as high shows that fimbriae bearing plays an important role in widespread and resistant strains, especially in recurrent UTIs such as in our study. In addition, hemolysin capability was also a virulence factor in recurrent UTIs (p<0.01). In addition, the sensitivity of the strains to the antimicrobials appeared in the following order; imipenem 93%, norfloxacin 89%, ciprofloxacin 85%, netilmicin 80%, amikacin 78%, ceftriaxone 74%, gentamicin 72%, nitrofurantoin 71%, ampicillin-sulbactam 60%, amoxicillin-clavulanate 58%, Trimethoprim/sulfamethoxazole 45%, ampicillin 35%.

Adult↗

In vitro antimicrobial activity of diethyldithiocarbamate and dimethyldithiocarbamate against methicillin-resistant Staphylococcus.

Staphylococcus aureus has appeared which is highly resistant to both methicillin and aminoglycosides. Current therapy involves long-term intravenous therapy of vancomycin. Since vancomycin is currently the only drug used to treat these patients, there is a need to develop additional antimicrobial therapy. The in vitro antimicrobial effect of the metal chelator, diethyldithiocarbamate (DDTC) and its structural analog dimethyldithiocarbamate (DMTC) were investigated. Both DDTC and DMTC were effective against S. aureus including methicillin-resistant S. aureus (MRSA). By agar diffusion, DDTC at 10 micrograms per disk produced zone sizes of 12 to 21 mm and at 100 micrograms per disk produced zone sizes of 26 to 34 mm against MRSA. The DMTC produced slightly greater zone sizes against MRSA of 16 to 24 mm and 24 to 37 mm for 10 micrograms per disk and 100 micrograms per disk, respectively. The minimum inhibitory concentration (MIC) for DMTC against MRSA was 6 micrograms per ml. Both DDTC and DMTC were also effective against enterococci, Proteus mirabilis, Klebsiella pneumoniae, Klebsiella oxytoca, Escherichia coli, Enterobacter cloacae, Enterobacter aerogenes, Salmonella species, Serratia marcescens and Citrobacter freundii at 100 micrograms per disk. The MICs of DMTC for Klebsiella pneumoniae, Klebsiella oxytoca, Escherichia coli, Salmonella and Citrobacter freundii were approximately 128 micrograms per ml while the MICs for Proteus vulgaris, Proteus mirabilis, Pseudomonas aeruginosa and Serratia marcescens was greater than or equal to 256 micrograms per ml. In addition, DMTC was synergistic with gentamicin against MRSA and coagulase-negative staphylococcus species, Enterobacter cloacae, Klebsiella pneumoniae and Pseudomonas aeruginosa. Additive and synergistic effects of DMTC were displayed with gentamicin against S. aureus including methicillin-resistant S. aureus.(ABSTRACT TRUNCATED AT 250 WORDS)

Anti-Bacterial Agents↗

Molecular epidemiology of ceftiofur-resistant Escherichia coli isolates from dairy calves.

Healthy calves (n = 96, 1 to 9 weeks old) from a dairy herd in central Pennsylvania were examined each month over a five-month period for fecal shedding of ceftiofur-resistant gram-negative bacteria. Ceftiofur-resistant Escherichia coli isolates (n = 122) were characterized by antimicrobial resistance (disk diffusion and MIC), serotype, pulsed-field gel electrophoresis subtypes, beta-lactamase genes, and virulence genes. Antibiotic disk diffusion assays showed that the isolates were resistant to ampicillin (100%), ceftiofur (100%), chloramphenicol (94%), florfenicol (93%), gentamicin (89%), spectinomycin (72%), tetracycline (98%), ticarcillin (99%), and ticarcillin-clavulanic acid (99%). All isolates were multidrug resistant and displayed elevated MICs. The E. coli isolates belonged to 42 serotypes, of which O8:H25 was the predominant serotype (49.2%). Pulsed-field gel electrophoresis classified the E. coli isolates into 27 profiles. Cluster analysis showed that 77 isolates (63.1%) belonged to one unique group. The prevalence of pathogenic E. coli was low (8%). A total of 117 ceftiofur-resistant E. coli isolates (96%) possessed the bla(CMY2) gene. Based on phenotypic and genotypic characterization, the ceftiofur-resistant E. coli isolates belonged to 59 clonal types. There was no significant relationship between calf age and clonal type. The findings of this study revealed that healthy dairy calves were rapidly colonized by antibiotic-resistant strains of E. coli shortly after birth. The high prevalence of multidrug-resistant nonpathogenic E. coli in calves could be a significant source of resistance genes to other bacteria that share the same environment.

Animals↗

Characterization of Staphylococcus aureus isolates recovered from bovine mastitis in Rio de Janeiro, Brazil.

Phenotypic characteristics, antimicrobial susceptibility, and genetic relationships were analyzed in 107 Staphylococcus aureus isolates recovered from cows with subclinical mastitis in Southeastern Brazil. Thirteen different biochemical patterns were detected among isolates. A predominant pattern represented by about 54% of the isolates was distributed among several herds. Isolates of distinct phenotypic profiles were also detected within a herd. Susceptibility to ampicillin, cefotaxime, cephalotin, chloramphenicol, erythromycin, gentamycin, kanamycin, nitrofurantoin, norfloxacin, ofloxacin, oxacillin, penicillin, rifampin, sulfamethoxazole-trimethoprim, tetracycline, trimethoprim, and vancomycin, determined by the disk diffusion method, was observed in 44.9% of isolates. On the other hand, 55.1, 7.4, and 2.8% of the strains were resistant to ampicillin/penicillin, tetracycline, and erythromycin, respectively. Genetic diversity was analyzed by pulsed-field gel electrophoresis using SmaI as the restriction enzyme. All isolates could be typed by pulsed-field gel electrophoresis, which identified 16 types and 24 subtypes. Type A and its subtypes comprised 54.2% of all isolates and were recovered from 6 of the 9 herds analyzed. Other types and subtypes were also found in multiple herds. Although multiple types and subtypes were found within a specific herd, a predominant type was frequently observed.

Animals↗

Antibiotic susceptibility profiles of new probiotic Lactobacillus and Bifidobacterium strains.

The antimicrobial susceptibilities and presence of plasmids in four new probiotic lactic acid bacteria (LAB) strains, Lactobacillus rhamnosus HN001 (DR20) HN067, Lactobacillus acidophilus HN017 and Bifidobacterium lactis HN019 (DR10), were determined. Resistance to 18 commonly used antibiotics was assessed by disk diffusion. The three Lactobacillus strains had similar antibiotic susceptibility profiles to those of Lactobacillus plantarum strain HN045 and two commercial probiotic Lactobacillus strains, GG and LA-1. The B. lactis strain HN019 had a similar profile to three commercial probiotic B. lactis strains (Bb12, HN049 and HN098). All 10 strains were sensitive to the Gram-positive spectrum antibiotics erythromycin and novobiocin, the broad-spectrum antibiotics rifampicin, spectinomycin, tetracycline and chloramphenicol and the beta-lactam antibiotics penicillin, ampicillin and cephalothin. By contrast, most strains were resistant to the Gram-negative spectrum antibiotics fusidic acid, nalidixic acid and polymyxin B and the aminoglycosides neomycin, gentamicin, kanamycin and streptomycin. All three L. rhamnosus strains (HN001, HN067 and GG) were resistant to vancomycin and several strains were also resistant to cloxacillin. Of the four new probiotic strains, only L. rhamnosus HN001 contained plasmids; however, a plasmid-free derivative of HN001 had the same antibiotic susceptibility profile as the parent strain.

Anti-Bacterial Agents↗

Occurrence of extended spectrum beta-lactamase enzymes in clinical isolates of Klebsiella species from Harar region, eastern Ethiopia.

Extended Spectrum beta-Lactamases (ESBLs) producer and multidrug resistant Klebsiella spp. are becoming a major nosocomial pathogen globally. There are no documented reports yet on the occurrence of ESBL enzymes in Klebsiella spp. species from Ethiopia. This study was undertaken to isolate and determine the occurrence of ESBLs and multi-drug resistant Klebsiella spp. in different clinical samples obtained from patients. A cross-sectional survey was conducted in four different hospitals of Harar region (Hiwot Fana, Misrak-Arbegnoch, Police and Army) from December 2003 to February 2004. Three hundred and eighty four clinical specimens (202 sputum, 164 urine and 18 pus) were collected from patients admitted in different wards. Antimicrobial susceptibilities were performed on 57 clinical isolates by standard disk diffusion procedures against eight antimicrobial agents. The ESBLs detection was made by using cefotaxime and ceftazidime alone and in combination with clavulanate. A total of 57 (15%) Klebsiella spp. were isolated from 384 patients. Of the 57 isolates, 33 (58%) were from sputum, 18 (31.5%) from urine and 6 (10.5%) from pus. Of the 57 Klebsiella spp., 54 (94.7%) were identified as K. pneumoniae and 3 (5.3%) as K. oxytoca. Resistance was found against cephalosporins [cefotaxime (39.0%), cefoxitin (39.0%), ceftazidime (40.0%), ceftriaxone (40.0%), cephalothin (42.0%)], chloramphenicol (70.0%), gentamicin (61.0%) and trimethoprim-sulphamethoxazole (65.0%). Analyzed Klebsiella isolates were characterized also by a high degree of multi-resistance (67.0%). In 19/57 (33.3%) of the Klebsiella isolates, ESBL production was detected. Rates of detection of ESBL producers were 42.1, 26.3, 26.3 and 5.3% in Hiwot-Fana, Misrak-Arbegnoch, Police and Army hospitals, respectively. Multi-drug resistant isolates were more prevalent among the ESBLs producers (95.0%) than non-producers (53.0%) (p=0.24). In conclusion, our results show that awareness of ESBL production by Klebsiella spp. is clinically important. In the absence of infection control measures, ESBL producing organisms readily pass horizontally from patient to patient. These strains also transiently colonize the hands of hospital staff members, thereby facilitating patient-to-patient transmission of the organism.

Cross Infection↗

Antimicrobial susceptibility of udder pathogens isolated from dairy herds in the west littoral region of Uruguay.

A total of 522 strains belonging to streptococci, enterococci and staphylococci isolated from sub-clinical and clinical cases of bovine mastitis from the west littoral region of Uruguay were analysed for their susceptibility to several antimicrobial agents. The susceptibility patterns were studied by agar disk diffusion methods (ADDM) and broth micro-dilution to determine the minimum inhibitory concentration (MIC). The concentration that inhibits 90% (MIC90) of the analysed strains reported in micrograms per millilitre, for Staphylococcus aureus were > 8, 8, < or = 0.5, < or = 4, < or = 1, < or = 0.5, > 64, < or = 0.25, 0.5, < or = 1 and < or = 1 to penicillin, ampicillin, oxacillin, cephalotin, gentamicin, erythromycin, oxitetracycline, enrofloxacin, trimethoprim/sulfamethoxazole, neomycin, and clindamycin, respectively. Coagulase-negative staphylococci (CNS) had different values for penicillin (4) and ampicillin (2), while the other antimicrobial agents had the same MIC90 values as reported for S. aureus. The MIC90 values for streptococci were 0.12, 0.25, < or = 4, 16, < or = 0.25, 0.5, 0.25 for penicillin, ampicillin, cephalotin, gentamicin, erythromycin, oxytetracycline and trimethoprim-sulfamethoxazole, whereas MIC90 for enterococci were 4, 4, 4, < or = 0.5, 2, > 8 for penicillin, ampicillin, gentamicin, erythromycin, oxytetracycline and trimethoprim-sulfamethoxazole, respectively. Of 336 strains of S. aureus, 160 (47.6%) were resistant to penicillin. For 41 CNS strains, 10 (27%) presented penicillin-resistance. All the streptococcal strains were susceptible to penicillin, while 3 (7%) of the 43 enteroccocal strains were resistant. Non significant statistical differences were found between the results obtained by ADDM and broth micro-dilution for classifying bacterial isolates as susceptible or resistant according to the National Committee of Clinical Laboratory Standards.

Animals↗

In vitro antimicrobial activities and spectra of U-100592 and U-100766, two novel fluorinated oxazolidinones.

Two new fluorinated oxazolidinones, U-100592 and U-100766, were evaluated against more than 659 gram-positive and -negative organisms and compared with glycopeptides, erythromycin, clindamycin, clinafloxacin, and chloramphenicol. U-100592 and U-100766 were usually equally potent, but the MICs at which 90% of the isolates are inhibited (MIC90s) of U-100592 for some staphylococci and enterococci were slightly lower than those of U-100766 (1 versus 2 micrograms/ml). The MIC90 of U-100592 and U-100766 for oxacillin-resistant Staphylococcus aureus was 2 micrograms/ml, the same as observed for oxacillin-susceptible strains. The oxazolidinone MICs for other Staphylococcus spp. were < or = 2 micrograms/ml (MIC50, 0.5 to 1 microgram/ml). All enterococci were inhibited by < or = 4 and < or = 2 micrograms of U-100592 and U-100766 per ml, respectively. Against 152 vancomycin-resistant enterococci (five species), both compounds had a narrow range of MICs (0.25 to 2 micrograms/ml) and a MIC90 of 1 microgram/ml. Corynebacterium jeikeium, Bacillus spp., and all tested streptococci were inhibited (< or = 4 micrograms/ml). Members of the family Enterobacteriaceae and other gram-negative bacilli were not susceptible (MIC50, > 64 micrograms/ml) to either oxazolidinone. Three potencies of U-100592 and U-100766 disks were tested (5, 15, and 30 micrograms), and acceptable correlations (r = 0.81 to 0.90) with the measured MICs were observed. Best discrimination of the tentatively susceptible organisms (MICs, < or = 4 micrograms/ml) was demonstrated with the 30-micrograms disk concentration. The oxazolidinones demonstrated a dominant bacteristatic action. These oxazolidinones (U-100592 and U-100766) appear promising for treatment of gram-positive organisms that demonstrate resistance to contemporary therapeutic agents.

Acetamides↗

[Blood cultures: use of presumptive antiobiograms].

Mortality associated to bacteremia varies between 20 and 40% depending upon several factors, such as focus of infection, microorganism, host conditions, etc. It has also been documented that mortality may double when the patient does not receive antibiotic treatment to which the microorganism is susceptible. The objective of our work has been to determine the correlation between disk diffusion antibiogram according to NCCLS guidelines, from isolated colonies, and the one performed directly from the blood culture flask. During 1996, in the Institute of Cardiology and Cardiovascular Surgery (ICYCC) in Buenos Aires City, 81 episodes of bacteremia were studied. In every case, an antibiogram was carried out: 1) from the bottle: a- Directly (D), harvesting 20 microliters in Mueller Hinton agar, b- Diluted (d), previous centrifugation and Gram staining to adjust turbidity equivalent to 0.5 Mc Farland; 2) from isolated colonies, according to NCCLS guidelines. There were almost no major errors, except with two strains of Enterobacter cloacae versus cephalotin. The diluted method was not so convenient to read inhibition zones, especially with staphylococci. With gram-positive bacteria, the main problems appeared in the direct method with erythromycin, oxacillin and ciprofloxacin because of minor errors. With gram-negative bacteria, major errors were observed in the direct method, mainly with piperacillin (7%) and to a lesser extent with piperacillin tazobactam (2%). Except for imipenem, trimethoprim sulfamethoxazoie and cefotaxime, all antimicrobial agents presented minor errors with both methodologies. Based upon the high rate of minor errors, we consider it is important to confirm results obtained with the standard technique (NCCLS), considering as presumptive those results from the blood culture bottles (D and d).

Anti-Bacterial Agents↗

Frequency of isolation of shigella serogroups/serotypes and their antimicrobial susceptibility pattern in children from slum areas in Karachi.

OBJECTIVES: To assess the frequency of serogroups and serotypes, as well as the antimicrobial susceptibility pattern of Shigella species isolated from known cases of diarrhoea and dysentery from Karachi, Pakistan. METHODS: A cross-sectional study was conducted between January 2002 and March 2003 at Aga Khan University on stool samples received from children with diarrhoea and dysentery from four low socio-economic areas (Sultanabad, Rehri Goth, Hijrat and Sherpao colony) of Karachi. Stool samples yielding growth of Shigella species, were further identified for serotypes by slide agglutination. Antibiotics susceptibility was performed by Kirby Bauer disk diffusion method. RESULTS: Out of 4688 stool samples received, 193 (4.1%) were positive for Shigella species. Shigella flexneri was the predominant serogroup (58%) followed by Shigella sonnei (16%), Shigella boydii (15%) and Shigella dysentriae being the least common (11%). A number of serotypes were isolated in each serogroup, 8 serotypes in S. flexneri, 8 serotypes in S. dysenteriae, 9 serotypes in S. boydii, and Phase 1 & 2 were found in S. sonnei. 17% isolates remained non-serotypeable. All isolates were susceptible to Ofloxacin and Ceftriaxone, high rate of resistance was observed in Cotrimoxazole (87.75%) and Ampicillin (55.5%). Emerging resistance against Nalidixic acid (39%) was observed. CONCLUSION: Shigella still accounts for a significant proportion of bacillary dysentery in many tropical and subtropical countries. Serotype identification can help in devising strategies such as development of effective vaccine for controlling this problem. Increasing antibiotic resistance against commonly prescribed drugs signify that treatment options have become difficult in cases of severe dysentery.

Ampicillin↗

Emergence of a Novel, Phenotypically Difficult-to-Detect Vancomycin-Resistant Enterococcus faecium Clone (ST117/CT7799).

A significant increase of vancomycin-resistant Enterococcus faecium (VREfm) infections was observed in South-Eastern Austria since 2024. The prolonged outbreak is caused by a novel vanB-VREfm clone (ST117/CT7799, "VREfmstyr"). This study characterizes the atypical difficult-to-detect resistance phenotype and assesses the genomic relatedness of the isolates. Patient and outbreak characteristics were investigated including whole genome sequencing of the isolates. Sensitivity of broth microdilution (BMD), gradient tests (GT), disk diffusion (DD), and automated susceptibility testing (VITEK2) was compared. The performance of commercial screening media was evaluated. From sporadic detections in early 2024 case numbers began to rise during the year. In 30/31 (97%) of all cases, intra-hospital transmission was considered likely and an association with invasive procedures was identified in most cases. Core genome multilocus sequence typing revealed only six allelic differences between VREfmstyr isolates collected in a 12-month period, all belonging to the E. faecium ST117/CT7799 lineage. BMD detected vancomycin resistance (MIC&#x2009;>&#x2009;4&#x2009;mg/L) in no more than 16/31 (52%) of isolates after 24&#x2009;h incubation, while GT and DD misclassified all isolates. Only prolonged incubation improved the performance of these assays. VITEK2 analysis, however, correctly classified all 31 isolates. Of four commercially available VRE-screening agars, only one was capable of detecting VREfmstyr after 24&#x2009;h incubation. The emergence and clonal dissemination of VREfm ST117/CT7799 reveals a serious diagnostic gap as commonly used diagnostic algorithms fail to reliably detect this resistance phenotype. Our findings should help to further evaluate the true geographical distribution and clinical significance of this novel VREfm clone.

Enterococcus faecium↗

[The prevalence of methicillin-resistant Staphylococcus aureus phagotype 95 in the Hospitales Vall d'Hebron of Barcelona].

BACKGROUND: In our hospital endemic methicillin resistant Staphylococcus aureus (MRSA) has been documented since 1971, with epidemic and interepidemic periods. During these years phage groups I, I-III, and non-typable were found by the international set of phages Phage group 95 (F95) was unusual between 1986 (when phage typing was first available) and 1991, with prevalence of 5.2% (mean), and 100% of sensibility to methicillin. In November 1991 appeared the first MRSA F95 strain, and its prevalence has been increasing until 1997. MATERIAL AND METHODS: We have studied 133 strains of MRSA F95 isolated from 87 patients, 39 of them hospitalized in the General Hospital (HG), 38 in Traumatology Hospital (HT) and 8 in the Children's Hospital (HI). Two of these patients had successive stancies in HG and HT. Antimicrobial susceptibility was determined by the disk diffusion method and microdilution to check oxacillin resistance. Moreover these method we have maked: detection of mecA, phage typing with the international set of phages and study of the PGFE patterns by digestion of chromosomic DNA with Smal. RESULTS: The percentage of methicillin resistance in F95 strains was increased since the appear of the first strain between 8.3% in 1991 to a maximum of 76.9% in 1995, we had a descens to 13.7% in 1996 but 1997 can back to augment it to 72.5%. MICs for oxacillin of these strains were low (< or = 64 mg/l to 87.4% of strains), and all of them were mecA positive, 78.1% of them were resistant to macrolides, 96.5% to tobramycin and 84.9% to quinolones, but only 10.5% to gentamicin, 4.7% were resistant to cotrimoxazol, 1.2% to fosfomycin and 2.5% to rifampin. All of them were sensible to doxycycline, and vancomycin. The pulse field gel electrophoresis showed 7 restriction patterns in MRSA F95, 73.8% of strains correspond to one of them (B), spreading from the spinal cord injury unit and prevalent in HT; and 10.8% to another (C), the first that appear, spreading from the neurosurgical unit and with high prevalence in HG. 6.9% has pattern J a B subtype that appear in broth HG and HT. Pattern E is prevalent in HI it was spread from neonatology unit. CONCLUSIONS: The emergence in a Center with endemic resistance of new strains of MRSA, not all of them of the same clone, with characteristic resistance pattern to antibiotics and in convivence with other phage groups is one demonstration of genetic variability of SAMR in our entorn.

Bacteriophage Typing↗

Validity of nalidixic acid screening in fluoroquinolone-resistant typhoid salmonellae.

OBJECTIVE: To validate the screening of low-level fluoroquinolone resistance in typhoid salmonellae by using nalidixic acid (30 mg) disk providing an acceptable zone of inhibition. DESIGN: Quasi-experimental study. PLACE AND DURATION OF STUDY: The Department of Microbiology, Armed Forces Institute of Pathology, Rawalpindi, Pakistan from July 2002 to June 2003. MATERIALS AND METHODS: Antimicrobial susceptibility of 225 clinical isolates of S. typhi (n=126) and S. paratyphi A (n=99) against nalidixic acid and ciprofloxacin was determined by the modified Kirby-Bauer disk diffusion and agar dilution techniques of NCCLS. The relationship between the zone sizes and the MICs of the two quinolones was plotted in the form of scattergrams and nalidixic acid MICs and zone of inhibition sizes were correlated with those of ciprofloxacin by regression analysis. RESULTS: One hundred and ninety-five isolates were nalidixic acid-susceptible (MIC <16 microg/mL) and approximately 30 were nalidixic acid-resistant (MIC >32 microg/mL). All the nalidixic acid-susceptible isolates had ciprofloxacin MIC of <0.064 microg/mL. Among the nalidixic acid-resistant isolates approximately 20 had ciprofloxacin MIC > or =0.125 microg/mL and approximately 10 had ciprofloxacin MIC < or =0.03-0.064 microg/mL. The diameter of inhibition zone around a 30 mg nalidixic acid disk of nalidixic acid-resistant isolates was < or =13 mm (range 6-16 mm, mean 10.3 mm + SD 3.5 mm), while among nalidixic acid-susceptible isolates it ranged from 14 to 30 mm (mean 23.8 mm + SD 2.2 mm). The diameter of inhibition zone around a 5mg ciprofloxacin disk of nalidixic acid-resistant isolates ranged from 26 to 35 mm (mean 29.8 mm + SD 3.1 mm), while in nalidixic acid-susceptible isolates it ranged from 32 to 42 mm (mean 36.6 mm + SD 1.9 mm). With ciprofloxacin MIC > or =0.125 microg/mL taken as a breakpoint, a zone of <or =33 mm around a 5 microg ciprofloxacin disk to detect low susceptibility strains had a sensitivity of 100% and a specificity of 82%. Screening for nalidixic acid resistance (inhibition zone diameter of < or =13 mm) in isolates with ciprofloxacin MIC 0.125 microg/mL using a 30 microg nalidixic acid disk yielded a sensitivity of 100% and a specificity of 95%. CONCLUSION: Screening for nalidixic acid resistance with a 30 microg nalidixic acid disk is a reliable and cost-effective method for detection of low-level fluoroquinolone resistance in typhoid salmonellae.

Anti-Infective Agents↗

Antimicrobial susceptibility patterns and characterization of clinical isolates of Staphylococcus aureus in KwaZulu-Natal province, South Africa.

BACKGROUND: Antimicrobial resistance of Staphylococcus aureus especially methicillin-resistant S. aureus (MRSA) continues to be a problem for clinicians worldwide. However, few data on the antibiotic susceptibility patterns of S. aureus isolates in South Africa have been reported and the prevalence of MRSA in the KwaZulu-Natal (KZN) province is unknown. In addition, information on the characterization of S. aureus in this province is unavailable. This study investigated the susceptibility pattern of 227 S. aureus isolates from the KZN province, South Africa. In addition, characterization of methicillin-sensitive S. aureus (MSSA) and MRSA are reported in this survey. METHODS: The in-vitro activities of 20 antibiotics against 227 consecutive non-duplicate S. aureus isolates from clinical samples in KZN province, South Africa were determined by the disk-diffusion technique. Isolates resistant to oxacillin and mupirocin were confirmed by PCR detection of the mecA and mup genes respectively. PCR-RFLP of the coagulase gene was employed in the characterization of MSSA and MRSA. RESULTS: All the isolates were susceptible to vancomycin, teicoplanin and fusidic acid, and 26.9% of isolates studied were confirmed as MRSA. More than 80% of MRSA were resistant to at least four classes of antibiotics and isolates grouped in antibiotype 8 appears to be widespread in the province. The MSSA were also susceptible to streptomycin, neomycin and minocycline, while less than 1% was resistant to chloramphenicol, ciprofloxacin, rifampicin and mupirocin. The inducible MLSB phenotype was detected in 10.8% of MSSA and 82% of MRSA respectively, and one MSSA and one MRSA exhibited high-level resistance to mupirocin. There was good correlation between antibiotyping and PCR-RFLP of the coagulase gene in the characterization of MRSA in antibiotypes 1, 5 and 12. CONCLUSION: In view of the high resistance rates of MRSA to gentamicin, erythromycin, clindamycin, rifampicin and trimethoprim, treatment of MRSA infections in this province with these antibacterial agents would be unreliable. There is an emerging trend of mupirocin resistance among S. aureus isolates in the province. PCR-RFLP of the coagulase gene was able to distinguish MSSA from MRSA and offers an attractive option to be considered in the rapid epidemiological analysis of S. aureus in South Africa. Continuous surveillance on resistance patterns and characterization of S. aureus in understanding new and emerging trends in South Africa is of utmost importance.

Anti-Bacterial Agents↗