Mercury Resistance and R Plasmids in Escherichia coli Isolated from Clinical Lesions in Japan

The mercury and antibiotic resistance of 338 strains of Escherichia coli isolated from hospital patients was determined. Resistance to mercury was found in 58.6% of the isolates. The frequencies of resistance to streptomycin (Sm), tetracycline (Tc), chloramphenicol (Cm), kanamycin (Kan), cephaloridine (Cer), and gentamicin (Gm) were 66.3, 60.3, 56.5, 42.9, 32.1, and 1.5%, respectively. Among the above, 198 mercury- and antibiotic-resistant isolates were selected and tested for their ability to transfer the resistance to susceptible strains of E. coli K-12 and Klebsiella pneumoniae JK5. R plasmids carrying mercury resistance were demonstrated in 89.9% of the mercury-resistant strains of E. coli. Furthermore, R(Hg,Sm,Tc,Cm) plasmids were demonstrated most frequently, followed by R(Hg,Sm,Tc,Cm,Kan), R(Hg,Cm,Kan), and R(Hg,Sm,Tc,Cm,Kan,Cer) plasmids.     

Drug Resistance and Distribution of R Factors in Salmonella Strains

Drug resistance and the distribution of R factors in Salmonella strains were surveyed using 1,980 strains isolated in Japan from 1955 to 1973. Resistances were mostly restricted to sulfanilamide (SA), tetracycline (TC), and streptomycin (SM), and combinations thereof. The demonstrated frequency of strains resistant to chloramphenicol (CM) was very low as compared with that in Escherichia coli and Shigella strains. In relation to resistance to TC, CM, SM and SA, the frequency of isolation of single resistance was the highest, followed by triple, double, and quadruple resistance in that order. Low frequency of isolation of quadruple resistance was due to the low frequency of CM resistance in Salmonella strains and differed from the E. coli or Shigella group. R factors with single TC resistance was most common, followed by those with TC,SM,SA; SM,SA; TC,CM,SM,SA; and single (SM and SA) resistance, in that order. Kanamycin and ampicillin resistance was unusual and mostly transmissible.     

Large Drug Resistance Virulence Plasmids of Clinical Isolates of Salmonella enterica Serovar Choleraesuis

Salmonella enterica serovar Choleraesuis generally causes systemic human salmonellosis without diarrhea, and therefore, antimicrobial treatment is essential for such patients. The drug resistance information on this organism is thus of high value. Serovar Choleraesuis usually harbors a virulence plasmid (pSCV) of 50 kb in size. Of the 16 clinical isolates identified to be serovar Choleraesuis, all except one harbored a pSCV and seven of them carried a pSCV of more than 125 kb in size. A pSCV was defined as a plasmid carrying spvC and characteristic deletions detected by PCR and by DNA-DNA hybridization (for the former criterion). The results of PCR, restriction fragment profiles, and Southern DNA-DNA hybridizations of the profiles all indicated that such larger pSCVs were derived from the 50-kb plasmid recombined with non-pSCVs found in some clinical isolates. Fifteen of the 17 strains, including a laboratory strain, were then tested for drug resistance against 16 antibiotics with E-test and the dilution method. The laboratory strain, which harbored a 50-kb pSCV and a 6-kb non-pSCV, was resistant only to sulfonamides (SUL), and its resistance gene, sulII, checked with PCR and DNA-DNA hybridization, was located on the 6-kb non-pSCV. All 14 clinical strains were resistant to multiple drugs. Of the 14, 7 were resistant to SUL, and the resistance gene was located on a plasmid. The sulII gene, but not bla(TEM-1), was carried only on the 6-kb non-pSCV. Of the remaining six large plasmids, three of 90 kb, two of 136 kb, and one of 140 kb, the last three were pSCVs and carried the other SUL gene (sulI) and the bla(TEM-1) gene. The six strains were also resistant to trimethoprim-sulfamethoxazole. None of the 50-kb pSCVs carried resistance genes. These drug resistance genes on the large pSCVs were apparently also acquired through recombination.     

Emergence of multidrug-resistant Salmonella enterica serovar typhi in Korea

A chloramphenicol-resistant strain of Salmonella enterica serovar Typhi was first noted in Korea in 1992, when a resistant isolate was detected in a returned traveler. Continued isolation of multidrug-resistant (MDR) strains thereafter in other settings prompted a retrospective analysis of laboratory records and phenotypic and genotypic analyses of 12 chloramphenicol-resistant isolates. Among these, one isolate was resistant only to chloramphenicol, and the other isolates were also resistant to ampicillin and co-trimoxazole. MDR was transferred by conjugation from 9 of the 11 isolates. PCR showed that all isolates had an incompatible group HI1 plasmid, and oriT was detected in 10 isolates, which included strains with an unsuccessful transfer of resistance. All of the ampicillin-resistant isolates had a beta-lactamase band of pI 5.4 and bla(TEM) alleles. A PCR amplicon from an isolate showed that the sequences were identical to those of bla(TEM-1), suggesting that all isolates had a TEM-1 beta-lactamase. All isolates had class 1 integrons: 10 isolates had integrons of ca. 1.2 kb with dhfr7 gene cassettes, and 1 isolate had an integron of ca. 2.3 kb with aacA4 and bla(OXA-1)-like gene cassettes. The pulsed-field gel electrophoresis patterns of 7 of 11 MDR isolates were identical and indistinguishable from those reported for isolates in India and Indonesia. In conclusion, some of the MDR strains in Korea are related to those in other Asian countries. Susceptibility testing became necessary for selection of antimicrobial agents for the optimal treatment of patients with the emergence of MDR Salmonella serovar Typhi in Korea.     

Chelocardin-Inducible Resistance in Escherichia coli Bearing R Plasmids

Two plasmid-linked tetracycline resistance characters, tet A and tet B, were distinguishable in part, according to the level of resistance they conferred to minocycline (<3 μg/ml for tet A; >6 μg/ml for tet B). Escherichia coli K-12 strains that harbored the tet B character were also resistant to tetracycline but susceptible to chelocardin. In such tet B strains, subinhibitory concentrations of tetracycline could induce resistance to chelocardin as well as to otherwise inhibitory concentrations of tetracyclines. Chelocardin itself was ineffective as an inducer and therefore could be used to select constitutively resistant mutants. E. coli K-12 strains harboring the tet A character were also resistant to tetracycline and susceptible to chelocardin; tetracycline did not induce resistance to chelocardin in these strains.     

Chloramphenicol Resistance Plasmids in Escherichia coli Isolated from Diseased Piglets

The plasmids in 19 chloramphenicol-resistant Escherichia coli strains of three pig pathogenic antigen types were studied in conjugation and transduction experiments. The plasmids had identical resistance patterns: streptomycin, spectinomycin, sulfonamides, and chloramphenicol (Sm, Sp, Su, Cm) and belonged to IncFII. One plasmid carried ampicillin resistance in addition. Restriction enzyme analysis of the deoxyribonucleic acid from five of the plasmids originating from the same herd showed that their digestion patterns with EcoRI were indistinguishable. EcoRI cleaved the deoxyribonucleic acid of a sixth plasmid from the same herd and displayed nine of the ten bands of the other five plasmids plus an additional six. It appears that the five plasmids with identical restriction patterns have a common origin and may be copies of the same plasmid from which the sixth may have developed. Four strains carried two plasmids each. In two of these strains, a plasmid with a tetracycline marker (Tc), or possibly the tetracycline marker alone, recombined frequently with the Sm Sp Su Cm plasmid without destroying any known function of the latter. The possibility that Tc is carried on a translocation sequence is discussed.     

R Factors in Strains of Salmonella typhi and Shigella dysenteriae 1 Isolated During Epidemics in Mexico: Classification by Compatibility

All 17 Salmonella typhi strains tested from the epidemic in Mexico carried R factors of compatibility group H, conferring resistance to chloramphenicol, streptomycin, tetracycline, and sulfonamides. Some S. typhi strains carried, in addition, non-conjugative, ampicillin resistance plasmids and R factors of the I or A-C complex. All 20 Shigella dysenteriae 1 strains tested of epidemic origin carried O-group R factors. Ampicillin resistance in S. dysenteriae 1 was not proved to be plasmid borne. R factors of group H were not identified in any of the tested Mexican isolates other than S. typhi, but R factors of group O were identified in Escherichia coli, Shigella flexneri, and one strain of S. typhi, as well as in the epidemic S. dysenteriae. An R factor was identified which seemed to have two compatibility specificities, groups Iomega and O.     

Antimicrobial Resistance and R Factors in Salmonella Isolated in California (1971–1972)

The antimicrobial resistance of 2,246 strains of Salmonella isolated from humans in California was determined. Resistance to one or more of the 12 antimicrobial agents tested was found in 32% of the isolates. Salmonella typhimurium strains represented 31% of the serotypes isolated; 49% of these strains were resistant. Fifty-one percent of S. heidelberg, 42.5% of S. newport, and 40% of S. saint paul strains were resistant. Seventy-seven percent of all resistant serotypes were resistant to two or more of the antimicrobial agents tested. R factors were demonstrated in 70% of the multiply resistant strains. Resistance of the Salmonella strains to one or more of the 12 antimicrobials tested and the frequency of resistance to tetracycline and ampicillin were significantly greater than resistance reported in other studies performed in this country. Chloramphenicol resistance was encountered in 33 (1.5%) of the Salmonella isolates; 19 of these were S. typhi demonstrating a pattern of resistance to streptomycin, sulfisoxazole, tetracycline, and chloramphenicol and carrying an R factor capable of transferring the complete pattern of resistance.     

Decrease of Drug Resistance in Salmonella in The Netherlands

Since 1974, tetracycline resistance in salmonellae of human and porcine origin has decreased nation-wide in The Netherlands. This decrease has coincided with the ban on incorporation of tetracycline in animal feeds.     

Salmonella enterica serovar typhi strains isolated in Korea containing a multidrug resistance class 1 integron

Six strains of Salmonella enterica serovar Typhi which were resistant to ampicillin, chloramphenicol, trimethoprim-sulfamethoxazole, streptomycin, tetracycline, and gentamicin were isolated in Korea. This multidrug resistance was transferred by a conjugative plasmid of about 50 kb. The plasmid harbored a class 1 integron, which included six resistance genes, aacA4b, catB8, aadA1, dfrA1, aac(6')-IIa, and the novel blaP2, in that order. All of the isolates showed the same-size plasmids and the same ribotyping patterns, which suggests a clonal spread of these multidrug-resistant isolates.     

临床分离金黄色葡萄球菌的耐药性分析

目的分析金黄色葡萄球菌(SAU)临床分离株的耐药特性,为临床合理用药提供参考依据。方法细菌鉴定应用VITEK 32全自动细菌鉴定分析系统,采用K-B法对临床分离金黄色葡萄球菌菌株进行药物敏感性试验;头孢西丁和苯唑西林纸片扩散法检测MRSA;双纸片扩散法检测诱导型克林霉素耐药。结果 216株SAU中耐甲氧西林金黄色葡萄球菌(MRSA)130株,占60.2%;MRSA对青霉素G、氨苄西林100.0%耐药;对万古霉素、利奈唑烷100.0%敏感;对氯霉素耐药率为8.5%;对呋喃妥因耐药率为14.6%;对大环内酯类、喹诺酮类、氨基糖苷类抗菌药物耐药率均较高(61.5%～90.0%),且存在多药耐药;MRSA耐药率普遍高于MSSA。苯唑西林纸片扩散法较头孢西丁纸片扩散法判断MRSA的漏检率为4.6%;克林霉素诱导耐药检出率为56.3%。结论临床分离金黄色葡萄球菌耐药性已十分严重;头孢西丁纸片扩散法是检测MRSA简便、可靠的方法;临床微生物室应常规检测金黄色葡萄球菌红霉素对克林霉素的诱导耐药性,以指导临床正确使用抗菌药物。     

R Plasmids in Corynebacterium xerosis Strains

Plasmids coding for resistance to chloramphenicol, erythromycin, kanamycin, streptomycin, and tetracycline have been found in strains of Corynebacterium xerosis isolated from patients with otitis media.     

某院2009～2011年尿培养中细菌分布及耐药性分析

目的监测和分析本院2009～2011年尿培养分离病原菌的种类、分布及耐药状况,指导临床合理运用抗菌药物。方法采用VITEK 2Compact系统对尿培养阳性标本分离菌株进行鉴定及药敏分析,应用WHONET 5.6软件分析中段尿培养标本所分离病原菌的分布及药敏情况。结果 3 571例尿培养标本中共分离出1 046株病原菌,阳性率为29.3%,其中革兰阴性菌601株,占57.5%,尤以大肠埃希菌最为常见;革兰阳性菌261株,占24.9%,以葡萄球菌和肠球菌为主;真菌184株,占17.6%。病原菌对各种抗菌药物耐药差异性较大,表现为多药耐药。结论本院尿路感染病原菌种类复杂多样、耐药程度严重,需及时监测病原菌的菌群种类分布和耐药变迁,以指导临床合理、规范地使用抗菌药物。     

Class 1 Integron-Borne Multiple-Antibiotic Resistance Carried by IncFI and IncL/M Plasmids in Salmonella enterica Serotype Typhimurium

The presence and genetic content of integrons were investigated for 37 epidemiologically unrelated multiple-drug-resistant strains of Salmonella enterica serotype Typhimurium from humans. All isolates were resistant to ampicillin, chloramphenicol, kanamycin, streptomycin, sulfonamides, and trimethoprim, as well as to tetracycline and/or nalidixic acid; 20% of them were also resistant to gentamicin and amikacin. Three different class 1 integrons (In-t1, In-t2, and In-t3) were identified by Southern blot hybridization, PCR, and DNA sequencing, and these integrons were found to carry the aadB, catB3, oxa1, aadA1a, aacA4, and aacC1 gene cassettes. Integrons In-t1 (aadB and catB3) and In-t2 (oxa1 and aadA1a) were both located on a conjugative IncFI plasmid of 140 kb. In-t3 (aacA4, aacC1, and aadAIa) was located on an IncL/M plasmid of 100 kb which was present, in association with the IncFI plasmid, in gentamicin- and amikacin-resistant isolates. Despite the extensive similarity at the level of the antibiotic resistance phenotype, integrons were not found on the prototypic IncFI plasmids carried by epidemic Salmonella strains isolated during the late 1970s. The recent appearance and the coexistence of multiple integrons on two conjugative plasmids in the same Salmonella isolate are examples of how mobile gene cassettes may contribute to the acquisition and dissemination of antibiotic resistance.Bacterial resistance to antimicrobial agents is a serious problem worldwide, and understanding of the molecular basis of how resistance genes are acquired and transmitted may contribute to the creation of new antimicrobial strategies (47). One efficient mechanism for the acquisition and dissemination of resistance determinants is their transmission through mobile genetic elements. It has been proposed that promiscuous plasmids, conjugative transposons, and transposons carried by conjugative plasmids are responsible for the horizontal spread of resistance genes throughout bacteria (9). Recently, naturally occurring gene expression elements called “integrons” have been described as vehicles for the acquisition of resistance genes carried by mobile elements (14, 26, 46). These structures have also been found to be involved in the genetic reassortment of resistance determinants frequently observed in multiple-antibiotic-resistant bacterial pathogens (5).Three classes of integrons have been identified. Class 1 integrons are prevalent among clinical isolates and are composed of two conserved regions, the 5′CS and the 3′CS regions, surrounding an interposed variable region (14, 16). The variable region contains gene cassettes for antibiotic resistance integrated, all in the same orientation, at the attI site. Coordinate expression of the gene cassettes is driven by the tandemly arranged P_ant and P2 promoters (39). The 5′CS region of class 1 integrons contains the intI1 gene, which encodes the type 1 integrase protein, which is responsible for site-specific insertion and excision of gene cassettes (5). As a consequence of this activity, integrons exist in a large variety of forms with respect to the number, type, and order of the inserted genes. Each gene cassette includes an open reading frame and a recombination site known as the “59-base element” located downstream of each coding sequence (15). The 3′CS contains the sul1 and the qacEΔ1 genes, which confer resistance to sulfonamides and to quaternary ammonium compounds, respectively (33, 36). Gene cassette arrays similar to those of the class 1 integrons were observed for the transposon Tn7 and its close relatives, forming a second class of integrons. A putative defective integrase gene (intI2), whose product is 40% identical to that of intI1, is located in the distal portion of Tn7 (11). Recently, a third class of integron has been identified, and the putative integrase (intI3), located at the 5′ of the bla_IMP cassette, is 61% identical to the intI1 integrase (31).The occurrence of integrons among clinical bacterial isolates is under investigation, and recent studies report a widespread distribution of these elements among multiple-antibiotic-resistant nosocomial bacteria (18, 20, 34).Salmonella enterica serotype Typhimurium is one of the more frequent agents of bacterial enteritis worldwide (29). This serotype has often been described as being resistant to multiple drugs, most commonly showing resistance to ampicillin (Ap), chloramphenicol (Cm), streptomycin (Sm), sulfonamides (Su), and tetracycline (Te). The R types R-ApCmSmSuTe and R-ApSmSuTe have been associated with the phage types DT104 and DT193, respectively (17, 23, 48).In the study described in this report, we investigated the genetic determinants for antibiotic resistance of S. enterica serotype Typhimurium strains isolated from epidemiologically unrelated pediatric patients with gastroenteritis. Our results indicate that the multiple-drug resistance phenotype is determined by integrons carrying an unusually wide repertoire of resistance gene cassettes. Up to three types of integrons located on two conjugative plasmids were identified. The quality, number, and organization of the resistance genes were then determined.     

Distribution of R Plasmids Among the O-Antigen Types of Escherichia coli Isolated from Human and Animal Sources

The O-antigen types of 600 independently isolated Escherichia coli strains from human feces have been determined, and the types have been related to the antibiotic resistance patterns of the strains. The relative abundance of each O-antigen type differed in the susceptible and resistant series of strains. The majority (86%) of the resistant strains carried R plasmids. Resistant E. coli (20.3%) were found associated with O-antigen types 8, 9 and 101, whereas the susceptible strains covered a wide range of O-antigen types. Examination of 174 resistant strains isolated from calf feces also showed a prevalence of O-antigen types 8, 9, 101 (24.1%), and it seems probable that strains expressing these three O-antigen types commonly carry R plasmids in the alimentary tracts of man and calves. The number of strains not typeable with the O sera available were similar in the human (12.5%) and the calf (11.5%) series. There are no grounds for distinguishing “human” from “calf” E. coli on the basis of their O-antigen reactions.     

IncI1 Plasmids Carrying Various blaCTX-M Genes Contribute to Ceftriaxone Resistance in Salmonella enterica Serovar Enteritidis in China

Resistance to extended-spectrum β-lactams in Salmonella, in particular, in serotypes such as Salmonella enterica serovar Enteritidis that are frequently associated with clinical infections, is a serious public health concern. In this study, phenotypic characterization of 433 clinical S. Enteritidis strains obtained from a nationwide collection of the Chinese Center for Disease Control and Prevention during the period from 2005 to 2010 depicted a trend of increasing resistance to ceftriaxone from 2008 onwards. Seventeen (4%) of the strains were found to be resistant to ceftriaxone, 7% were found to be resistant to ciprofloxacin, and 0.7% were found to be resistant to both ciprofloxacin and ceftriaxone. Most of the ceftriaxone-resistant S. Enteritidis strains (15/17) were genetically unrelated and originated from Henan Province. The complete sequence of an IncI1 plasmid, pSE115, which belonged to a novel sequence type, was obtained. This 87,255-bp IncI1 plasmid was found to harbor a bla_CTX-M-14 gene in a novel multidrug resistance region (MRR) within the tra locus. Although the majority of strains were also found to contain conjugative IncI1 plasmids with a size similar to that of pSE115 (∼90 kb) and harbor a variety of bla_CTX-M group 1 and group 9 elements, the novel MRR site at the tra locus in pSE115 was not detectable in the other IncI1 plasmids. The findings from this study show that cephalosporin resistance in S. Enteritidis strains collected in China was mainly due to the dissemination of IncI1 plasmids carrying bla_CTX-M, resembling the situation in which IncI1 plasmids serve as major vectors of bla_CTX-M variants in other members of the Enterobacteriaceae.