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PCR-based identification of bacteria associated with endodontic infections.

PCR primers that target the bacterial 16S rRNA genes (or the tuf gene for the genus Enterococcus) were used to identify 10 putative bacterial pathogens in root canals with necrotic pulp. In addition, the associations of these microorganisms with symptoms and a history of diabetes mellitus were investigated. Microbial samples from the root canals of 24 teeth with necrotic pulp were included in the study. PCR with universal bacterial primers identified bacterial DNA in 22 specimens; the remaining 2 specimens were from intact teeth that had been traumatized 6 months prior to treatment. PCR with specific primers showed that preoperative symptoms were significantly associated with the presence of Streptococcus spp. (P < 0.001 by chi-square analysis). There was also a nonsignificant trend for symptoms to be associated with Fusobacterium nucleatum and Porphyromonas gingivalis (odds ratio, >2) and for diabetes mellitus to be associated with P. gingivalis and Porphyromonas endodontalis (odds ratio, >2). Cloning and sequencing of the universal PCR product in one specimen revealed the presence of an organism related to the genus Olsenella, which has not previously been described in endodontic infections.

Bacteria↗

In vitro antimicrobial activity of propolis and Arnica montana against oral pathogens.

Arnica and propolis have been used for thousands of years in folk medicine for several purposes. They possess several biological activities such as anti-inflammatory, antifungal, antiviral and tissue regenerative, among others. Although the antibacterial activity of propolis has already been demonstrated, very few studies have been done on bacteria of clinical relevance in dentistry. Also, the antimicrobial activity of Arnica has not been extensively investigated. Therefore the aim here was to evaluate in vitro the antimicrobial activity, inhibition of adherence of mutans streptococci and inhibition of formation of water-insoluble glucan by Arnica and propolis extracts. Arnica montana (10%, w/v) and propolis (10%, w/v) extracts from Minas Gerais State were compared with controls. Fifteen microorganisms were used as follows: Candida albicans--NTCC 3736, F72; Staphylococcus aureus--ATCC 25923; Enterococcus faecalis--ATCC 29212; Streptococcus sobrinus 6715; Strep. sanguis--ATCC 10556; Strep. cricetus--HS-6; Strep. mutans--Ingbritt 1600; Strep. mutans--OMZ 175; Actinomyces naeslundii--ATCC 12104, W 1053; Act. viscosus OMZ 105; Porphyromonas gingivalis; Porph. endodontalis and Prevotella denticola (the last three were clinical isolates). Antimicrobial activity was determined by the agar diffusion method and the zones of growth inhibition were measured. To assess cell adherence to a glass surface, the organisms were grown for 18 h at 37 degrees C in test-tubes at a 30 degree angle. To assay water-insoluble glucan formation, a mixture of crude glucosyltransferase and 0.125 M sucrose was incubated for 18 h at 37 degrees C in test-tubes at a 30 degree angle. Arnica and propolis extracts (20 microl) were added to these tubes to evaluate the % of inhibition of cell adherence and water-insoluble glucan formation. The propolis extract significantly inhibited all the microorganisms tested (p < 0.05), showing the largest inhibitory zone for Actinomyces spp. The Arnica extract did not demonstrate significant antimicrobial activity. Cell adherence and water-insoluble glucan formation were almost completely inhibited by the propolis extract at a final concentration of 400 microg/ml and 500 microg/ml, respectively. The Arnica extract showed slight inhibition of the adherence of the growing cells (19% for Strep. mutans and 15% for Strep. sobrinus) and of water-insoluble glucan formation (29%) at these same concentrations. Thus, the propolis extract showed in vitro antibacterial activity, inhibition of cell adherence and inhibition of water-insoluble glucan formation, while the Arnica extract was only slightly active in those three conditions.

Actinomyces↗

A new rapid automated method for the detection of Listeria from environmental swabs and sponges.

Many food and meat processors test environmental swabs and sponges to confirm the absence of Listeria spp. Spectral pattern changes in a liquid growth medium, resulting from esculin hydrolysis by Listeria in contaminated swabs and sponges, were automatically monitored by the BioSys instrument in a semifluid layer (SFL). The blackening of SFL in modified MOX broth resulted in sharply declining curves, which were easily detected by the instrument. The instrument detected all nine strains of Listeria monocytogenes tested. None of the gram negative organisms (Proteus, Escherichia coli, Pseudomonas, Citrobacter and Yersinia) were detected by the system, nor were most gram positive organisms, including Bacillus, Streptococcus, and Lactobacillus strains, Staphylococcus aureus, Enterococcus faecium and E. faecalis, which hydrolyze esculin, produced black colonies on PALCAM and Oxford media and were also detected in the system. A total of 122 sponges and swabs collected at food processing plants were evaluated by this method. Of these, 99 were negative, and 11 were positive. L. innocua was the dominant Listeria species in these environmental samples. Good correlation was obtained between numbers of Listeria and detection times of esculin hydrolysis: 1000 CFU/swab were detected in 10-13 h, whereas 1-10 CFU/swab were detected in less than 22 h. The total assay time was 26 h.

Animals↗

Infective endocarditis not related to intravenous drug abuse in HIV-1-infected patients: report of eight cases and review of the literature.

OBJECTIVES: To add to the limited information on infective endocarditis (IE) not related to intravenous drug abuse (IVDA) in HIV-1-infected patients. METHODS: We have reviewed the characteristics of eight cases of IE in non-IVDA HIV-1 infected patients diagnosed in our institution between 1979 and 1999 as well as cases in the literature. RESULTS: All our patients were male, and the mean age was 44 years (range 29-64). HIV-1 risk factors were: homosexuality in five, heterosexuality in two, and the use of blood products in one. HIV stage C was found in six cases, and the median (range) CD4 cell count was 22/microL (4-274 cells/microL). IE was caused by Enterococcus faecalis in three cases, staphylococci in two cases, and Salmonella enteritidis, viridans group streptococci and Coxiella burnetii in one case each. Three patients acquired IE while in the hospital. All IE cases involved a native valve, and underlying valve disease was found in three patients. The aortic valve was the most frequently affected (five cases). Two patients underwent surgery, with a good outcome, and one patient died. Fourteen cases of IE not related to IVDA in HIV-1-infected patients were found in the literature review. The most common causative agents were Salmonella spp. and fungi (four cases each). Two patients had prosthetic valve IE, and the mitral valve was the most frequently affected (10 cases). The remaining clinical characteristics and the outcome were similar to those in the present series. CONCLUSIONS: IE not related to IVDA is rare in HIV-1-infected patients. In more than half of the cases, IE develops in patients with advanced HIV-1 disease. A wide etiologic range is found, reflecting different clinical and environmental conditions. None of the patients who underwent surgery died, and the overall mortality rate was not higher than in non-HIV-1-infected patients with IE.

AIDS-Related Opportunistic Infections↗

Antimicrobial susceptibility of quinupristin/dalfopristin tested against gram-positive cocci from Latin America: results from the global SMART (GSMART) surveillance study.

Gram-positive cocci are important causes of both nosocomial and community-acquired infections, and antimicrobial resistance among these pathogens has become an important problem worldwide. Since resistance among these organisms can vary substantially by geographic location, we conducted a multicenter surveillance study with isolates from five Latin American countries (15 medical centers). Quinupristin/dalfopristin (formerly RP-59500) is a novel streptogramin combination with focused activity against Gram-positive cocci, many exhibiting emerging resistance. The in vitro activity of quinupristin/dalfopristin and 12 other antimicrobial agents were evaluated against 1,948 strains including Staphylococcus aureus (747 strains), coagulase-negative staphylococci (CoNS;446 strains), enterococci (429 strains), and various Streptococcus spp. (326 strains). Oxacillin resistance was observed in 41% of S. aureus (MIC, e; 13 mm) and 40% of CoNS (MIC, e; 18 mm). Vancomycin, teicoplanin, and quinupristin/dalfopristin (MIC(90), 0.25 - 1 mg/ml) remained effective against all strains, but cross-resistance was high among other tested drugs. The quinupristin/dalfopristin MIC(50) for Streptococcus pneumoniae and other streptococci was only 0.5 mg/ml (13% to 28% were penicillin-resistant; 12% to 22% were macrolide-resistant). Enterococci demonstrated variable inhibition by quinupristin/dalfopristin depending upon identification and the susceptibility testing method used. The demonstrated quinupristin/dalfopristin activity against Enterococcus faecium was confirmed, but potential species identification errors with various commercial systems continue to confuse susceptibility statistics, even though some strains of E. faecium confirmed by PCR-based or other molecular identification techniques did have quinupristin/dalfopristin MICs of >e; 4 microg/ml. Most important, glycopeptide-resistant enterococci are rapidly emerging in Latin America, and quinupristin/dalfopristin appears active against many of these isolates as well as having potency against nearly all staphylococci and streptococci tested at e; 16 mm. Comparisons to GSMART results from other continents show nearly identical quinupristin/dalfopristin activity for each Gram-positive species tested. These results define the role of quinupristin/dalfopristin in Latin American medical centers and provide a benchmark for future in vitro comparisons.

Anti-Bacterial Agents↗

Increasing prevalence of vancomycin-resistant enterococci, and cefoxitin-, imipenem- and fluoroquinolone-resistant gram-negative bacilli: a KONSAR study in 2002.

Continued antimicrobial resistance surveillance can provide valuable information for the empirical selection of antimicrobial agents for patient treatment, and for resistance control. In this 6th annual study for 2002, the susceptibility data at 39 Korean Nationwide Surveillance of Antimicrobial Resistance (KONSAR) hospitals were analyzed. Resistance rates of S. aureus were 67% to oxacillin, and 58% to clindamycin. The ampicillin and vancomycin resistance rates of E. faecium were 89% and 16%, respectively. To penicillin, 71% of S. pneumoniae were nonsusceptible. Resistance rates of E. coli were 11% to cefotaxime, 8% to cefoxitin, and 34% to fluoroquinolone, and those of K. pneumoniae were 22% to ceftazidime, and 16% to cefoxitin. Lowest resistance rates to cephalosporins shown by E. cloacae and S. marcescens were to cefepime, 7% and 17%, respectively. This is the first KONSAR surveillance, which detected imipenem-resistant E. coli and K. pneumoniae. To imipenem, 22% of P. aeruginosa and 9% of Acinetobacter spp. were resistant. Trends of resistances showed a slight reduction in MRSA and in penicillin- nonsusceptible S. pneumoniae, but an increase in ampicillin-resistant E. faecium. Ampicillin-resistant E. coli and H. influenzae remained prevalent. Compared to the previous study, amikacin- and fluoroquinolone- resistant Acinetobacter spp. increased to 60% and 62%, respectively. Ceftazidime- resistant K. pneumoniae decreased slightly, and imipenem- resistant P. aeruginosa and Acinetobacter spp., and vancomycin-resistant E. faecium increased. In conclusion, vancomycin-resistant E. faecium, cefoxitin-resistant E. coli and K. pneumoniae, and imipenem-resistant P. aeruginosa and Acinetobacter spp. increased gradually, and imipenem- resistant E. coli and K. pneumoniae appeared for the first time. Continued surveillance is required to prevent further spread of these serious resistances.

Anti-Bacterial Agents↗

[Clinical laboratory approach for estimating effective administrative dose of cefoperazone. Evaluation of disc susceptibility test and its interpretation system].

To interpret of the cefoperazone (CPZ) disc susceptibility test, a 4 category system is used in Japan, but a 3 category system is used in the U.S.A. and Europe. In the 4 category interpretation system of Showa CPZ disc the following classification is used: ( ) MIC less than or equal to 3 micrograms/ml, (++) MIC greater than 3 approximately 15 micrograms/ml, (+) MIC greater than 15 approximately 60 micrograms/ml, (-) MIC greater than 60 micrograms/ml. In the 3 category system the classification used is as follows: susceptible MIC less than or equal to 16 micrograms/ml, moderately susceptible MIC greater than 16 approximately 32 micrograms/ml, resistant MIC greater than 32 micrograms/ml, or susceptible MIC less than or equal to 32 micrograms/ml, moderately susceptible MIC greater than 32 approximately 64 micrograms/ml, resistant MIC greater than 64 micrograms/ml, depending on dose levels, 1 or 2 g. Reliability of the CPZ disc susceptibility test in estimating approximate MICs by classifying the test results into 4 categories was studied using discs containing 1, 2, 5, 10, 30 and 75 micrograms. The MICs were determined using the agar dilution method at an inoculum level of 10(6) CFU/ml. A good negative correlation was observed between inhibitory zone diameters and MICs, showing reliability of the test using these discs. The results obtained with discs containing 30 or 75 micrograms of CPZ were well categorized into the 4 groups mentioned above. Some strains of Pseudomonas aeruginosa and Enterococcus faecalis, however, showed false positive results. When different break points of inhibitory zone diameters than those used for other bacteria were used for P. aeruginosa, and E. faecalis was excluded from the test, an excellent correlations were obtained. With 30 or 75 micrograms discs, it was unable to subclassify strains against which MICs of CPZ were below 3 micrograms/ml. However, with discs containing 1 to 10 micrograms, it was possible to separate the strains against which MICs were less than 0.5 microgram/ml. The fact that most frequent values of MICs of CPZ against Escherichia coli, Klebsiella pneumoniae, Proteus spp., Haemophilus influenzae, Streptococcus pyogenes etc. were less than 0.5 microgram/ml supports the usefulness of low dose discs. According to recently ongoing concepts on the pharmacokinetics of antibiotics and their penetration into tissues and inflammatory fluids, serum protein binding appear to be one of the important determinants of drug distribution in the body. Only free, unbound drug molecules can readily pass through capillary pores into tissue fluids except into hepatic biliary system.(ABSTRACT TRUNCATED AT 400 WORDS)

Cefoperazone↗

Douglas-fir root-associated microorganisms with inhibitory activity towards fungal plant pathogens and human bacterial pathogens.

A microbial culture collection composed of 1820 bacterial strains, including 298 actinomycete strains, was established from the roots of Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) seedlings harvested from conifer nurseries and forest sites. Two hundred and thirty-four strains inhibited the growth of Fusarium, Cylindrocarpon, and (or) Pythium spp. in in vitro assays. A significantly greater proportion of bacterial strains from actinomycete genera exhibited antifungal properties compared with bacterial strains from nonactinomycete genera. Eighty-nine percent of identified inhibitory strains were Streptomyces, Streptoverticillium, Bacillus, Pseudomonas, or Burkholderia species. The actinomycete species were isolated almost exclusively from forest seedlings. Recovery of inhibitory strains representing 29 microbial species was enhanced using a variety of methods to isolate microorganisms from the roots of seedlings from nursery and forest sites. Bacterial strains (including actinomycete strains) with antifungal activity were tested for in vitro growth inhibition of six clinical human bacterial pathogens (Enterococcus faecalis, Staphylococcus aureus, Klebsiella pneumoniae, Escherichia coli, Proteus mirabilis, and Pseudomonas aeruginosa). Forty-eight percent of the tested strains inhibited one or more human pathogens, Inhibitory activity towards fungal and bacterial pathogens was strain specific, not species specific, and many inhibitory strains exhibited broad-spectrum activity. Strains with antifungal activity against several conifer root pathogens were also more likely to inhibit multiple species of clinical bacterial pathogens.

Actinomycetales↗

In vitro evaluation of BAY Y3118, a new full-spectrum fluoroquinolone.

BAY Y3118, 1-cyclopropyl-7-(2,8-diazabicyclo[4.3.0]non-8-yl)-6-fluoro-8- chloro-1,4-dihydro-4-oxo-3-quinolinecarboxylic acid hydrochloride, is a new fluoroquinolone with antibacterial activity against an expanded spectrum of species including Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Enterococcus faecium, Streptococcus pneumoniae, Streptococcus pyogenes, and also anaerobes such as Bacteriodes fragilis and Clostridium perfringens. MIC90s for S. aureus, S. epidermidis, E. faecalis, and S. pneumoniae clinical isolates were 0.125, 0.25, 0.125 and 0.25 micrograms/ml, respectively. Against methicillin- and/or quinolone-resistant S. aureus, MIC50 levels of BAY Y3118 were 10- to 100-fold lower than those of tosufloxacin, sparfloxacin, or ciprofloxacin. The potency of BAY Y3118 against all members of the Enterobacteriaceae generally was equal to or 2-fold greater than that of ciprofloxacin or tosufloxacin. BAY Y3118 was also highly active against Haemophilus influenzae, Moraxella catarrhalis, Acinetobacter spp. and Pseudomonas aeruginosa. Increasing inoculum concentrations had a minimal effect on MIC determinations. The drug was determined to be bactericidal based upon reference MBCs and time-kill curves. From the results presented here, it was concluded that this new compound surpasses other known 4-quinolones both in spectrum and activity and that its further evaluation by in vitro, in vivo, and clinical studies seems warranted.

Anti-Infective Agents↗

Association between Streptococcus faecalis urinary infections and graft rejection in kidney transplantation.

In the first month after transplantation 50% of 193 consecutive renal transplant recipients had bacteriuria. The most common organisms isolated were Streptococcus faecalis (34), Escherichia coli (28), Pseudomonas spp. (11), and staphylococcus (9). There was a significant correlation between infection with Str. faecalis and graft failure at one, three, and twelve months. This observation suggests that urinary infection with Str. faecalis may be associated with graft failure which is probably the result of immunological factors.

Adult↗

In-vitro antimycoplasmal activity of flurithromycin.

The in vitro activity of flurithromycin, a 14-membered macrolide drug, was found to be similar to that of erythromycin against 41 strains of Mycoplasma spp. and 100 strains of Ureaplasma urealyticum. All 28 strains of Mycoplasma hominis were uniformly resistant to both macrolides with MICs > 256 mg/L, U. urealyticum showed intermediate resistance with MIC50s of 0.5 and 1 mg/L for erythromycin and flurithromycin, respectively, whereas the ten strains of Mycoplasma pneumonia were susceptible to < or = 0.03 mg/L of both macrolides.

Enterococcus faecalis↗

Evaluation of the effect of 3 probiotics on experimental Escherichia coli enterotoxaemia in weaned piglets.

In this report the efficacy of 3 different probiotics (Bacillus cereus "toyoi", Lactobacillus spp. and Streptococcus faecium) was investigated. They were supplemented in the food of recently weaned piglets that were orally infected with E. coli O141 K85ab. Supplementation could not prevent mortality and clinical symptoms nor reduce the faecal excretion of hemolytic E. coli. The possible explanations for the unsatisfying results are various.

Animal Feed↗

Antimicrobial activity of doripenem (S-4661): a global surveillance report (2003).

The spectrum of activity and potency of doripenem, a broad-spectrum parenteral carbapenem currently in clinical development, was evaluated using 16 008 clinical bacterial isolates collected as part of an international surveillance project during 2003. Using reference broth microdilution methods, doripenem was found to be highly active against oxacillin-susceptible Staphylococcus aureus and coagulase-negative staphylococci (2705 and 297 isolates, respectively; MIC90s 0.06 mg/L), with a potency greater than that of other carbapenem antibiotics. Against enterococci (1474 isolates), with the exception of Enterococcus faecium, doripenem displayed modest activity (MIC50 4). Doripenem was among the most potent agents tested against Streptococcus pneumoniae, viridans group streptococci and beta-haemolytic streptococci (885, 140 and 397 isolates; MIC(90)s 0.5, 0.5 and 0.03 mg/L, respectively). For Enterobacteriaceae (> 6200 isolates), doripenem was four- to 32-fold more active than imipenem against wild-type isolates (MIC90s 0.03-0.5 mg/L). MIC90s for confirmed extended-spectrum beta-lactamase-producing Escherichia coli and Klebsiella pneumoniae (121 and 155 isolates; 0.06 and 0.12 mg/L, respectively) were two-fold higher than for wild-type isolates. Doripenem was also active against Citrobacter spp., Enterobacter spp. and Serratia spp. (MIC90s 0.06-0.25 mg/L), including ceftazidime-resistant isolates. Doripenem and meropenem were the most active agents among all beta-lactams against Pseudomonas aeruginosa (829 isolates; MIC50/90s 0.5/8 and 0.5/16 mg/L, respectively), whereas doripenem and imipenem were the most active agents against Acinetobacter spp. (155 isolates; MIC50/90s 0.5/4 and <or= 0.5/2 mg/L, respectively). Doripenem was slightly more potent (MIC50 2 mg/L) than ertapenem and imipenem (MIC50 4 mg/L), and had a potency similar to that of meropenem (MIC50 2 mg/L), against Burkholderia cepacia (20 isolates). Both Haemophilus influenzae (1824 isolates) and Moraxella catarrhalis (108 isolates), including beta-lactamase-positive isolates, were susceptible to doripenem (MIC90s 0.25 and 0.03 mg/L, respectively). Doripenem displays the favourable characteristics of other carbapenems, and appears to offer certain advantages in terms of potency, spectrum and beta-lactamase stability when compared with some carbapenems used currently to treat nosocomial infections.

Anti-Bacterial Agents↗

Leuconostoc mesenteroides subsp. mesenteroides FR52 synthesizes two distinct bacteriocins.

Mesenterocin 52, a bacteriocin produced by Leuconostoc mesenteroides subsp. mesenteroides FR52, was purified from producing cells by the adsorption-desorption method, combined with reverse-phase high-performance liquid chromatography. The elution profile revealed the presence of two inhibitory peaks of activity, each displaying different inhibitory spectra. Mesenterocin 52A possessed a broad inhibitory spectrum, including anti-Listeria activity, while Mesenterocin 52B was only active against Leuconostoc spp. The amino acid sequence and M(r) of Mesenterocin 52A appeared identical to the previously described Mesentericin Y105. In contrast, Mesenterocin 52B possessed a M(r) of 3446 Da, corresponding to 32 amino acids and a sequence that shared no homology with known bacteriocins: NH2-KGVLGWLSMASSALTGPQQPNSPWLAKIKNHK.

Amino Acid Sequence↗

Antimicrobial actions of calcium binding leucocyte L1 protein, calprotectin.

The calcium binding L1 protein was found to inhibit growth of blood culture isolates of Candida spp and cerebrospinal fluid isolates of Cryptococcus neoformans. Minimum inhibitory concentrations (MIC) were 4-128 mg/l, and concentrations 2-4 times the MIC were fungicidal. Blood culture isolates of Escherichia coli, Klebsiella spp, Staphylococcus aureus, and Staphylococcus epidermidis had MIC values of 64-256 mg/l. Antibacterial activity was strongly influenced by the nature of the culture medium. In view of the biological activity of L1, the name calprotectin is proposed to describe this antimicrobial protein with calcium binding properties.

Antifungal Agents↗

Identification and composition of the streptococcal and enterococcal flora of tonsils, intestines and faeces of pigs.

Streptococcus suis was the most frequent Streptococcus spp. in pig tonsils, followed by the beta-haemolytic porcine 'equisimilis' ecovar of Strep. dysgalactiae. The intestinal streptococcal flora was composed of Strep. bovis, Strep. hyointestinalis and Strep. suis. Many of these intestinal Strep. suis belonged to a beta-glucuronidase-negative biotype which is infrequent in lesions. Nearly half of the strains presumptively identified as Strep. alactolyticus produced acid from lactose. This species was not found in tonsils and intestines but was about equally prevalent as Strep. hyointestinalis in pig faeces and rectal swabs. Other streptococci were rare in this material. Enterococci were much less frequently identified than streptococci in tonsils and faeces. In intestinal samples Enterococcus faecalis, Ent. faecium, Ent. hirae and Ent. cecorum were most frequently found. In faeces Ent. faecium was the most prevalent enterococcus. The characteristics of the less well known species Strep. alactolyticus and Strep. hyointestinalis are described in detail, and guidelines for their differentiation from Strep. bovis and Strep. suis given.

Animals↗

Acquired macrolide resistance genes in pathogenic Neisseria spp. isolated between 1940 and 1987.

Seventy-six Neisseria gonorrhoeae isolates, isolated between 1940 and 1987, and seven Neisseria meningitidis isolates, isolated between 1963 and 1987, were screened for the presence of acquired mef(A), erm(B), erm(C), and erm(F) genes by using DNA-DNA hybridization, PCR analysis, and sequencing. The mef(A), erm(B), and erm(F) genes were all identified in a 1955 N. gonorrhoeae isolate, while the erm(C) gene was identified in a 1963 N. gonorrhoeae isolate. Similarly, both the mef(A) and erm(F) genes were identified in a 1963 N. meningitidis isolate. All four acquired genes were found in later isolates of both species. The mef(A) gene from a 1975 N. gonorrhoeae isolate was sequenced and had 100% DNA and amino acid identity with the mef(A) gene from a 1990s Streptococcus pneumoniae isolate. Selected early isolates were able to transfer their acquired genes to an Enterococcus faecalis recipient, suggesting that these genes are associated with conjugative transposons. These isolates are the oldest of any species to carry the mef(A) gene and among the oldest to carry these erm genes.

Anti-Bacterial Agents↗

Detection by PCR of the nim genes encoding 5-nitroimidazole resistance in Bacteroides spp.

A PCR method was developed for detection of the nim genes encoding 5-nitrolmidazole resistance in Bacteroides spp. Two PCR primers specific for nim genes were designed. They allowed amplification of a 458-bp fragment from all characterized plasmid- and chromosome-borne metronidazole resistance genes. The specificity of the method was tested with DNA from metronidazole-sensitive Bacteroides spp. strains and from other strains of unrelated species. Each DNA preparation was analyzed with and without an internal positive control to verify that the absence of PCR amplification product was not due to inhibition of the Taq polymerase inhibitors. By this technique, two newly discovered metronidazole-resistant clinical strains of Bacteroides fragilis were shown to harbor resistance genes undetectable by Southern blotting. In spite of the sequence divergence of the nim genes, the PCR method is thus suitable for epidemiological investigations. The amplification method also revealed that nim-related resistance genes were not present in either Streptomyces strain S6670, a natural producer of 2-nitroimidazole, or in Enterococcus faecalis strains, which have been suggested to possess metronidazole-inactivating enzyme.

Bacterial Proteins↗