Escenario actual de la resistencia a la colistina mediada por plásmidos en América Latina / Current scenario of plasmid-mediated colistin resistance in Latin America

Colistin resistance can occur by chromosomal mutations and by acquisition of plasmid-carrying determinants, mainly mcr-1. In the recent years, we have observed the out-burst of this resistance gene in our region. Due to the risk of the rapid dissemination of mcr-1, this finding has worried and alerted different actors from the health field and has become one of the most prolific topics. Our review compiles available reports of well-documented mcr-1-positive strains of Enterobacteriaceae, obtained from different samples in Argentina and other countries of Latin America. Furthermore, it addresses the association of mcr-1 with ESBL resistance markers and outlines the platforms involved in their dissemination.

La resistencia a la colistina puede ocurrir por mutaciones cromosómicas o por la adquisición de determinantes localizados en plásmidos, el principal es mcr-1. En los últimos años hemos observado la explosiva aparición de este gen de resistencia en nuestra región. Debido al riesgo que implica la rápida diseminación de mcr-1, este hallazgo ha preocupado y alertado a los diferentes actores del área de la salud, y se ha convertido en uno de los temas de investigación más importantes. La presente revisión compila los informes de aislamientos portadores de mcr-1 debidamente documentados en Enterobacteriaceae, obtenidos de diferentes muestras en Argentina y otros países de América Latina. Además, aborda la asociación de mcr-1 con otros marcadores de resistencia, como las BLEE, y describe las plataformas involucradas en su diseminación.

Resistance of Corynebacterium pseudotuberculosis in the Brazilian semiarid environment / Resistência de Corynebacterium pesudotuberculosis no ambiente do semiárido brasileiro

The semiarid northeast of Brazil contains a unique biome known as caatinga, with a maximum temperature of 40 ºC and a relativity humidity of 56%. The caatinga is characterized by a variety of plants, including Cereus jamacaru Dc (mandacaru), Poincianella microphylla Mart. ex G. Don (catingueira), Pilosocereus gounellei FAC Weber (xique-xique) and Mimosa tenuiflora (Willd.) Poir (jurema preta). Sheep and goat industries are economically strong in that region, despite the fact that caseous lymphadenitis is highly prevalent. The aim of the present study was to assess the survival and biofilm production of Corynebacterium pseudotuberculosis isolates in the environment and under controlled temperatures (28°C, 37°C and 42°C) under different surfaces (plants, soil, wood, wire and thorns). In addition, we investigated the effects of applying the disinfectants chlorhexidine, hypochlorite and quaternary ammonia in soil, tiles, wood and vegetation cover. Four strains of C. pseudotuberculosis were selected (two from goats and two from sheep) for inoculation according to their in vitro biofilm production. Adherence to microplates was used to assess the biofilm-forming ability of the bacteria. Lower survival rates were observed when isolates of C. pseudotuberculosis were subjected to a temperature of 42°C. In terms of caatinga biome plants, contamination of jurema-preta plants resulted in the lowest survival rates. The disinfectant quaternary ammonia promoted a lower inoculum survival in all surfaces. The disinfectants and the higher temperature contributed to the reduction of biofilm production in isolates of C. pseudotuberculosis. knowledge of these patterns is important for the establishment of disease control measures, given the questionable efficacy of the treatment and the immuno-prophylaxis of caseous lymphadenitis.(AU)

O semiárido nordestino do Brasil possui um bioma exclusivo, a caatinga, que apresenta temperatura máxima de 40oC e uma umidade relativa do ar de 56%. A caatinga é caracterizada por uma diversidade de plantas, entre elas Cereus jamacaru DC. (mandacaru), Poincianella microphylla Mart ex G. Don (catingueira), Pilosocereus gounellei F.A.C. Weber (xique-xique) e Mimosa tenuiflora (Willd.) Poir (jurema preta). A produção de ovinos e caprinos está em franca expansão, porém a linfadenite caseosa é uma enfermidade de alta prevalência na região. Objetiva-se com o presente estudo, verificar a sobrevivência e produção de biofilme em isolados de Corynebacterium pseudotuberculosis em temperaturas de 28oC, 37oC e 42oC, quando inoculado em superfícies de solo, madeira, arame e espinho e em plantas nas condições ambientais da caatinga. Além disto, foi verificado o efeito da aplicação dos desinfetantes clorexidine, hipoclorito e amônia quaternária sobre o solo, piso (lajota), madeira e vegetação de cobertura do solo. Foram selecionadas quatro amostras de C. pseudotuberculosis (dois caprinos e dois ovinos) para inoculação de acordo com a sua produção in vitro de biofilme. A adesão a microplacas foi utilizado para avaliar a capacidade de formação de biofilme das bactérias. As menores taxas de sobrevivência foram observadas quando isolados de C. pseudotuberculosis foram submetidos a uma temperatura de 42°C. Com relação as plantas do bioma caatinga, a contaminação na planta jurema-preta apresentou menores índices de sobrevivência. O desinfetante amônia quartenária promoveu uma menor sobrevivência do inóculo em todas as superfícies. Os desinfetantes e temperatura contribuíram para a redução na produção de biofilme nos isolados de Corynebacterium pseudotuberculosis. O conhecimento destes padrões é importante para o estabelecimento de medidas de controle da enfermidade, dada a eficiência questionável do tratamento e imunoprofilaxia da linfadenite caseosa.(AU)

Caracterización de cepas de listeria monocytogenes realizados a partir de queso fresco proveniente de diferentes zonas productoras costarricenses / Characterization of listeria monocytogenes isolates obtained from raw cheese samples acquired from different Costa Rican producer zones

En Costa Rica, cerca del 25 por ciento de la producción de leche nacional es utilizada en la elaboración de queso tierno no pasteurizado, y el consumo de este producto es aproximadamente de 4 a 5 kg anuales per cápita. Este alimento ha sido involucrado en brotes debidos a Listeria monocytogenes. Dado lo anterior, se aisló e identificó esta bacteria a partir de muestras de queso blanco no pasteurizado provenientes de dos zonas tradicionalmente productoras y expendedoras de dicho producto. Se recolectaron 110 muestras de queso a partir de las cuales se aislaron 27 cepas de L. monocytogenes. Las cepas fueron caracterizadas mediante pruebas bioquímicas y serológicas, además se les realizaron pruebas de susceptibilidad a los antibióticos, hemólisis en tubo e invasión en células Hela. El 85 por ciento de las cepas evaluadas fueron sensibles a todos los antibióticos analizados, no obstante, cuatro cepas (15 por ciento) presentaron patrones de resistencia a diversos agentes, incluyendo estreptomicina, kanamicina, cefalotina y tetraciclina. También, se encontraron patrones de resistencia múltiple. El 88,9 por ciento de los aislamientos estudiados fueron positivos para la prueba de hemólisis en tubo, y el 22,2 por ciento presentaron porcentajes de invasión iguales o superiores a la cepa de origen clínico usada como control. Cabe destacar que todas las cepas con capacidad de invasión fueron también susceptibles a todos los antibióticos usados. Los resultados encontrados ponen de manifiesto la presencia de L. monocytogenes en queso blanco de origen costarricense. También se evidencia un alto porcentaje de susceptibilidad a los antibióticos de uso común para los casos de listeriosis. Por otro lado, pone de manifiesto que el queso blanco puede ser transmisor de cepas con capacidad de invasión y por ende, potencialmente patógenas al hombre.

In Costa Rica, almost 25 percent of the national milk production is used for the elaboration of non pasteurized soft cheese, and the annual intake of this product is around 4-5 kg per capita. This product has been identified as the source of food borne outbreaks due to Listeria monocytogenes. Given that, the isolation and identification of this bacterium from non pasteurized soft cheese samples coming from two producer zones of Costa Rica was performed. 110 cheese samples were collected, from which 27 L. monocytogenes strains were isolated. These were characterized using biochemical and serological tests, also, susceptibility to common used antibiotics, test tube hemolysis and invasion in Hela cells trials were performed. 85 percent of the strains evaluated were sensible to all the antibiotics analyzed, nevertheless, four strains presented resistance to different agents, including streptomycin, kanamycin, cephalotin and tetracycline. Also, multiple resistance patterns were found. 88,9 percent of the studied isolates were positive for the test tube hemolysis trial; 22,2 percent presented invasion percentages higher than the clinical origin strain used as control. It is important to point out that all the invasive strains were completely susceptible to the antibiotics tested. The results found demonstrate the presence of L. monocytogenes in Costa Rican soft cheese samples. Also, demonstrate its high percent of susceptibility to common use antibiotics. Same time, invasion trials show that soft cheese may be a source of invasive and potentially pathogenic strains for human being.

Evaluación del rendimiento de la resistencia general aerobica en judokas de alto nivel / Evalution of the performance of the aerobic genral resitencia in hih-level judokas

Para la evaluación del rendimiento de la resitencia general aeróbica en judokas de alto nivel se propuso el TEst de Cooper, se sometieron a la prueba 40 judokas de sexo masculino de alta competencia de la Asociación de Judo La PAz, fueron seleccionados de acuerdo al tiempo de práctica y la edad. Las pruebas se realizarón en la pista atlética del estadio "Hernando Siles", donde en primer lugar se registró datos generales de los judokas, tambien se efectuó una evaluación física y un control de los signos vitales a cada uno de los atletas que se cumplieron el test de Cooper...

Caracterización de un brote de Shigella boydii 14 por primera vez en Cuba / Characterization of an aoutbreak of Shigella boydii 14 for the first time in Cuba

Se propuso la caracterización de 14 cepas de Shigella boydii 14 aisladas de pacientes con enfermedad diarreica aguda mediante sus plásmidos de resistencia y de las proteínas de la membrana externa presentes en ellas. Se realizó la determinación de la susceptibilidad antimicrobiana por el método de concentración mínima inhibitoria, la extracción de plásmidos R fue según Manaitis, los extractos proteicos de las cepas se obtuvieron según el método de Blaser modificado y las proteínas de la membrana externa fueron separadas por SDS-PAGE por el método de Laemmli. Se comprobó que las cepas resultaron resistentes a la ampicilina (100 porciento), la tetraciclina (70 porciento) y al cotrimoxazol (50 porciento), y sensibles al ácido nalidíxico y a la ciprofloxacina. Se observó la presencia de plásmidos al nivel de los 43; 23; 20; 5,6 y 1,2 kb. Las proteínas de la membrana externa y el perfil proteico demostraron diferencias con otras especies de Shigella. Este serotipo de Shigella se aísla por primera vez en Cuba y y sus características la hacen altamente patógena y de muy difícil diagnóstico, por lo que la caracterización de este brote es importante desde el punto de vista epidemiológico

Caracterización de un brote de Shigella boydii 14 por primera vez en Cuba / Characterization of on aoutbreak of shigella boydii 14 for the first time in Cuba

Se propuso la caracterización de 14 cepas de Shigella boydii 14 aisladas de pacientes con enfermedad diarreica aguda mediante sus plásmidos de resistencia y de las proteínas de la membrana externa presentes en ellas. Se realizó la determinación de la susceptibilidad antimicrobiana por el método de concentración mínima inhibitoria, la extracción de plásmidos R fue según Manaitis, los extractos proteicos de las cepas se obtuvieron según el método de Blaser modificado y las proteínas de la membrana externa fueron separadas por SDS-PAGE por el método de Laemmli. Se comprobó que las cepas resultaron resistentes a la ampicilina (100 porciento), la tetraciclina (70 porciento) y al cotrimoxazol (50 porciento), y sensibles al ácido nalidíxico y a la ciprofloxacina. Se observó la presencia de plásmidos al nivel de los 43; 23; 20; 5,6 y 1,2 kb. Las proteínas de la membrana externa y el perfil proteico demostraron diferencias con otras especies de Shigella. Este serotipo de Shigella se aísla por primera vez en Cuba y y sus características la hacen altamente patógena y de muy difícil diagnóstico, por lo que la caracterización de este brote es importante desde el punto de vista epidemiológico(AU)

Salmonella: determinaçäo de sorotipos e resistência a agentes antimicrobianos de cepas isoladas de carcaças de frango comercializadas na regiäo de Säo José do Rio Preto - SP / Salmonella: serotyping and antimicrobial susceptibility patterns of strains isolated from chickem carcasse samples commercializede in the Säo José do Rio Preto area - SP

De abril de 1995 a dezembro de 1996, foram estudadas 160 amostras de carcaças de frango, comercializadas na Regiäo de Säo José do Rio Preto - SP. Foram determinados os sorotipos e perfis de sensibilidade aos agentes antimicrobianos de todas as cepas de Samonella isoladas. Das carcaças analisadas, 54,38(por cento) estavam contaminadas por Samonella. Foram identificados 18 diferentes sorotipos, dentre os quais Samonella Enteritidis correspondeu a 59,77(por cento). Verificou-se que 13,79 (por cento) das cepas de Samonella apresentaram resistência aos agentes antimicrobianos testados. Os resultados obtidos demonstraram o alto índice de contaminaçäo por Samonella em carne de frango. Considerando o consumo de alimentos de origem animal, principalmente de aves, por um grande número de pessoas, este fato possivelmente tem propiciado a ocorrência de numerosos surtos por S. Entidades na nossa regiäo, nos últimos anos

Molecular epidemiology of antibiotic resistance of Salmonella enteritidis during a 7-year period in Greece.

A significant increase in the frequency of isolation of Salmonella enteritidis has been observed during recent years in Greece, parallelled by an increasing rate of resistance of this organism to antibiotics. A substantial proportion of ampicillin- and doxycycline-resistant isolates exhibited cross-resistance to drugs of other classes, such as sulfonamides and streptomycin. Isolates of human origin were overall less resistant than those of animal or food-feed origin. Indeed, strains associated with animal infections were characterized by the highest rates of resistance to several antibiotics. These phenotypic data were correlated with genotypic information concerning two distinct populations: isolates from all sources that were resistant only to ampicillin, the drug toward which resistance rates were highest, and a control group of sensitive isolates. Ampicillin resistance was due to a 34-MDa conjugative plasmid. DNA fingerprinting by macrorestriction of genomic DNA revealed two types, A and B, common to both ampicillin-resistant and -sensitive strains, with 80 to 90% of strains being of type A. However, a third type, C, was specific for the sensitive population, representing 17% of those strains. Therefore, although the majority of resistant isolates were genetically related to sensitive ones, there existed a susceptible clone which had not acquired any resistance traits.

Relationship between adhesion to intestinal Caco-2 cells and multidrug resistance in Klebsiella pneumoniae clinical isolates.

Klebsiella pneumoniae is an opportunistic gram-negative pathogen involved in outbreaks of nosocomial infections in intensive care units. Strains are resistant to multiple antibiotics, and 15 to 30% of them are also resistant to the broad-spectrum cephalosporins by the production of R plasmid-encoded extended-spectrum beta-lactamases. Because the gastrointestinal tracts of patients have been shown to be the reservoir for nosocomial strains of K. pneumoniae, we looked for a correlation between antibiotic resistance and adhesion of K. pneumoniae strains to intestinal cells. We investigated adhesion to the human intestinal epithelial Caco-2 cell line of 61 clinical K. pneumoniae strains isolated in hospitals in Clermont-Ferrand, France. None of the strains tested expressed the previously described adhesive factors CF29K and KPF-28. Adhesive properties were found for 42.6% of the strains tested (26 strains). Just 7.7% (2 strains) of the 26 strains producing only the chromosomally encoded SHV-1 beta-lactamase adhered to the Caco-2 cell line, whereas 68.5% (24 strains) of the 35 strains producing a plasmid-encoded beta-lactamase were adherent. All the adherent strains, and even the two strains producing only the SHV-1 enzyme, harbored at least one self-transmissible R plasmid. At variance for CAZ-1/TEM-5 or CAZ-5/SHV-4 beta-lactamase-producing K. pneumoniae strains, curing and mating experiments demonstrated that the self-transmissible R plasmids encoding the TEM-1, CTX-1/TEM-3, CAZ-2/TEM-8, CAZ-6/TEM-24, or CAZ-7/TEM-16 beta-lactamase were not involved in the adhesion of K. pneumoniae strains to intestinal epithelial cells. Nevertheless, there was an association of multiple antibiotic resistance, including resistance to extended-spectrum cephalosporins, and adhesive properties in K. pneumoniae clinical isolates.

[Impact of control measures on the course of an outbreak of methicillin-resistant Staphylococcus aureus]. / Impacto de las medidas de control sobre la evolución de un brote epidémico por Staphylococcus aureus resistente a la meticilina.

BACKGROUND: Outbreaks of nosocomial infection by methicillin resistent Staphylococcus aureus (MRSA) are a problem in many hospitals with the control measures to be adopted being controversial. An outbreak of MRSA in a 550-bed university hospital is herein described and the impact of the adopted control measures on the evolution of the epidemic in the general hospitalization area (GHA) was analyzed. PATIENTS AND METHODS: The adopted control measures in the GHA were: microbiologic surveillance, cutaneous isolation measures, treatment of nasal carrier, and the early discharge of the cases. Hand washing was reinforced and a study of carriers was carried out on detection of sporadic cases (not related to the ICU). A molecular study of 70 strains of MRSA was performed with analysis of total plasmids, plasmid restriction pattern and chromosomic DNA analysis by pulsed field gel electrophoresis (PFGE). RESULTS: From December 1990 to December 1993, 273 cases of MRSA were reported. One hundred seventy-two cases originated in the ICU and 101 cases in the GHA (sporadic cases). The incidence of MRSA in 1991-1993 was 13.6, 14.3, and 6.6% in the ICU and 0.17, 0.36, and 0.15% in the GHA, respectively. Molecular study of MRSA isolates (1991 and 1992) demonstrated two plasmid and two chromosomic patterns. The latter had a similarity coefficient > 0.90, probably belonging to the same "clone". CONCLUSIONS: Despite the control measures adopted in the GHA the outbreak of MRSA originated in the ICU thereafter extending to the GHA. The rates of colonization detected, however, remained stable during the 3 years studied. On the other hand, the observation of a single "clone", responsible for the epidemic, suggest that most of the sporadic cases were autoctonous and due to failure in fulfillment of the established norms.

Evolution of drug resistance in Salmonella panama isolates in Chile.

In a search for Salmonella isolates in the environment in Chile in 1975, drug-susceptible strains of Salmonella panama were recovered for the first time from river water and vegetables in the vicinity of Santiago. Two to 3 years later, antibiotic-resistant S. panama began to appear in a variety of sources (meat, animals, vegetables, etc.), giving rise to a human epidemic that involved the entire nation. Of 139 clinical isolates studied, 7 were drug susceptible, 11 were resistant only to nitrofurans, and 3 were streptomycin, spectinomycin, and nitrofuran resistant; none of these 21 isolates harbored plasmid DNA. Most isolates (n = 107) were resistant to nitrofurans (chromosomal) and to streptomycin, spectinomycin, sulfonamides, tetracycline, and mercuric and tellurite salts; this multidrug resistance was encoded on a 218-kb plasmid classified in a number of strains as being in the IncHI2 group. From 1982 to 1993, 11 isolates acquired an additional self-transferable plasmid coding for resistance to any one of ampicillin (61 kb), ampicillin and trimethoprim (65 kb), ampicillin, trimethoprim, streptomycin, and sulfonamides (71 kb), ampicillin, gentamicin, kanamycin, and tetracycline (120 kb), or a nontransferable plasmid of approximately 6 kb encoding resistance to ampicillin or kanamycin. With the exception of ampicillin or ampicillin and trimethoprim resistance, S. panama isolates from foodstuffs, mainly pork meat products, and animals had resistance patterns that were the same as those found in clinical specimens. Remarkably, strains from goats and goat cheese and from shellfish isolated in particular rural regions were either drug susceptible or resistant only to streptomycin-spectinomycin encoded on a mobile genetic element and to nitrofurans. The report describes the arrival of a susceptible S. panama strain, its spread all over the country, and the evolution of progressively complex resistance patterns.

Genetic analysis of tetracycline-resistant plasmids in enteropathogenic Escherichia coli isolated from patients in Nigeria.

Genetic analysis of antibiotic-resistant plasmids from 102 serologically defined strains of enteropathogenic Escherichia coli from Nigeria was carried out. All the isolates were screened for susceptibility to antibiotics, and 47 were found resistant to tetracycline. A total of 138 plasmids was isolated by agarose gel electrophoresis. Transformation and conjugation experiments showed that 57.4% of the resistant strains carried R-plasmids ranging in sizes from 2 to 46 x 10(6) daltons. Plasmid-determined resistance to tetracycline, ampicillin and streptomycin was found. Restriction endonuclease analysis of three of the commonest plasmids: p1679, p529 and p1479 revealed relatedness with respect to function and structure. The DNA segment on which TcR gene is located on each of them was identified by cloning into the vector plasmid pGL101. The recombinant plasmids pOADI and pOAD2 gave full expression of TcR gene when transformed into E. coli DHI. Furthermore, the tetracycline-resistant strains were examined for their phenotypic behaviour with respect to tetracycline and its lipophilic analogs.

[Drug resistance and plasmid analysis in Pseudomonas aeruginosa].

One hundred and ninety seven strains of Pseudomonas aeruginosa were isolated from urine (59 strains), wound pus (60 strains), sputum (30 strains), stool (30 strains) and eye discharge (18 strains) at Kaohsiung medical College Hospital. These strains were serotyped with antisera against O antigens and tested with twelve different antimicrobial agents. The results showed that the most frequently isolated strains were serotype E (41.1%), followed by serotype B (20.3%), serotype F (10.7%) and serotype L (9.1%). In in vitro susceptibility testing, all isolated strains were resistant to chloramphenicol and tetracycline. Otherwise, these isolates were highly susceptible to ceftazidime (95.4%), enoxacin (89.3%) and piperacillin (87.8%). The isolates from urine exhibited more multiple drug resistance patterns than those of other specimens. When plasmid content was analysed from pseudomonas aeruginosa, only 15.2% (30/197) of isolates carried plasmids. By conjugation, transformation and mobilization experiments, it was shown that 13.3% (4/30) of plasmid carrying strains contained R plasmids.

Genetic and molecular R-plasmid analysis of Enterobacteriaceae hospital strains at Children's Hospitals of the former USSR.

R-plasmids from Enterobacteriaceae clinical strains, mainly Klebsiella and Serratia, isolated at different neonatal and children's hospitals of different cities of the former USSR for 10 years, were studied for their possible influence on the bacterial host phenotype. Hospital R-plasmids of stable inheritance persisted in hospitals from 2 to 7 years and were disseminated among strains of different genera (Klebsiella, Serratia, Enterobacter) and among different units. The data showed a possibility of long-term molecular rearrangements of R-plasmids in the hospital settings and an acquisition of genetic determinants encoding enterotoxin production. A novel R-plasmid encoding cytotoxicity to HEp-2 cells involved in two nosocomial outbreaks due to K. pneumoniae strains was reported. K. pneumoniae population heterogeneity was evaluated by using the plasmid parameters of strains. Their heterogeneity of a bacterial population was significantly lower during nosocomial outbreaks than in interepidemic periods.

Gentamicin resistance in clinical isolates of Escherichia coli encoded by genes of veterinary origin.

Seven (27%) of 26 gentamicin-resistant human clinical isolates of Escherichia coli were resistant to the veterinary aminoglycoside antibiotic apramycin. A gentamicin-resistant Klebsiella pneumoniae isolate from a patient infected with gentamicin/apramycin-resistant E. coli was also resistant to apramycin. DNA hybridisation studies showed that all gentamicin/apramycin-resistant isolates contained a gene encoding the enzyme 3-N-aminoglycoside acetyltransferase type IV (AAC[3]IV) that mediates resistance to gentamicin and apramycin in bacteria isolated from animals. Seven of the eight gentamicin/apramycin-resistant isolates were also resistant to the veterinary antihelminthic agent hygromycin B, a phenomenon observed previously in gentamicin/apramycin-resistant Enterobacteriaceae isolated from animals. Resistance to gentamicin/apramycin and hygromycin B was co-transferable in six of the isolates. Restriction enzyme analysis of plasmids in apramycin-resistant transconjugants derived from E. coli and K. pneumoniae isolates from the same patient were virtually identical, suggesting that inter-generic transfer of plasmids encoding apramycin resistance had occurred in vivo. These findings support the view that resistance to gentamicin and apramycin in clinical isolates of E. coli results from the spread of resistant organisms from animals to man, with subsequent inter-strain or inter-species spread, or both, of resistance genes on transferable plasmids.

[Mitral endocarditis caused by Staphylococcus aureus resistant to methicillin, aminoglucosides and rifampicin: description of 2 cases with fatal course]. / Endocarditis mitral por Staphylococcus aureus resistente a meticilina, aminoglucósidos y rifampicina: descripción de 2 casos de evolución fatal.

Two patients with infectious endocarditis (IE) by Staphylococcus aureus resistant to methicillin, aminoglucosides and rifampicin (SARMAR) acquired in hospital during the course of an epidemic outbreak of this microorganism in the Hospital Clínic i Provincial of Barcelona. Both patients had undergone surgery of the lower limbs. The entrance of the microorganism was the infection of the surgical wound, with bacteriemia, followed by mitral IE after a short time interval (20 days). Despite adequate treatment with vancomycin both patients died. The culture of mitral vegetation was positive for SARMAR in one. Analysis of the chromosomic DNA of all the isolations from the patients was identical and coincided with that of the SARMAR strains isolated in the epidemic outbreak of the hospital. The current situation of IE by SARMAR is reviewed and the therapeutic implications commented upon suggesting that treatment of this entity should simultaneously include the administration of vancomycin and phosphomycin or cotrimoxazole, with surgery being considered if infection persists.

The presence of high-molecular weight proteins with a strong affinity for cadmium in environmental Escherichia coli strains.

High molecular weight proteins, with a strong affinity for cadmium, were found in two environmental strains of Escherichia coli isolated from a wastewater treatment plant and were resistant up to 128 ppm of Cd+2. The fraction containing intracellular cadmium binding proteins was obtained by affinity chromatography and the single components of the same fraction were separated by SDS-gel electrophoresis in order to calculate molecular weights ranging from 48 to 89 kD. Plasmid analysis, carried out by agarose gel electrophoresis, and transformation experiments demonstrated that the plasmids, isolated from one of the strains which was resistant to tetracycline and streptomycin, are not related to the synthesis of cadmium-binding proteins.

Increased frequency of large R-plasmids in Klebsiella pneumoniae colonizing patients with spinal cord injury.

From 1978 to 1988 strains of gentamicin-susceptible (Gms) and gentamicin-resistant (Gmr) Klebsiella pneumoniae were saved from annual surveillance cultures of the perineal region of patients with spinal cord injury (SCI). Of 38 strains selected for further study (24 Gms and 14 Gmr), there were 23 different serotypes (two nontypable). Fourteen Gms as well as 14 Gmr strains displayed no common plasmid patterns, but all contained a large plasmid of 168-208 kb. Among the 14 Gmr strains, nine had large conjugative plasmids of approximately the same size (166-193 kb), which conferred to a susceptible Escherichia coli host an identical resistance pattern: ampicillin, chloramphenicol, gentamicin, piperacillin, trimethoprim-sulfamethoxazole, tetracycline, and tobramycin. Of the nine transconjugants, eight contained a single plasmid. One transconjugant contained a 168- and 80-kb plasmid. Restriction endonuclease digestion patterns of the R-plasmids revealed minimal similarity. We conclude that, during a 10-year period, different large R-plasmids have spread among multiple serotypes of K. pneumoniae in spinal cord injury (SCI) patients in one rehabilitation hospital. We hypothesize that other genes located on large, R-, and non-R-plasmids may confer an additional advantage for colonization by K. pneumoniae in SCI patients.

Enterotoxigenic drug resistant plasmids in animal isolates of Escherichia coli and their zoonotic importance.

A study was undertaken to determine how widely drug resistant enterotoxigenic plasmids are distributed among the animal isolates of Escherichia coli, and their potential for exchange. Thirty-one strains of E. coli isolated from metritis, septicaemia and diarrhoea of animals were tested for the production of enterotoxins. Thirteen strains were enterotoxin producers. Five produced heat-stable enterotoxins (ST), four heat-labile (LT) and four strains produced both (LT/ST) types of enterotoxins. Most of the enterotoxin producing E. coli strains were isolated from diarrhoeal sources, followed by septicaemia and metritis. Of the 13 enterotoxigenic E. coli, nine were resistant to various antibiotics which transferred their drug resistance partially or completely into an E. coli K-12 recipient. Only one strain of serogroup 032, isolated from equine metritis, transferred ampicillin, oxytetracycline and doxycycline resistance and heat-labile enterotoxin biosynthesis determinants en bloc. Plasmid DNA analysis of the exconjugants showed the presence of a 41.8 MDa conjugative plasmid. The presence of such autotransferable enterotoxin biosynthesis replicons in their unusual ecological niches such as metritis and septicaemia suggests the ubiquitous nature of enterotoxin genes which perhaps were selected and spread due to the presence of genes for drug resistance. The potential of cross-infection of human beings from animals and vice versa via such plasmids is discussed.

Comparison of antimicrobial resistant Escherichia coli in wild and captive Japanese serows.

The fecal Escherichia coli isolated from wild Japanese serows living in mountainous areas away from humans and those from captive serows kept in human areas were examined for antimicrobial resistance and the possession of transferable R plasmids. Of 874 E. coli strains isolated from 283 wild serows in 1980-1981, only 11 (1.3%) were resistant to at least one of 6 antimicrobial drugs; ampicillin, streptomycin, tetracycline, chloramphenicol, kanamycin and sulfadimethoxin. Seven (2.5%) individuals were found to carry resistant E. coli. To heighten the isolation frequency of drug-resistant strains, fecal samples of 244 wild serows in 1983-1984 were cultured directly onto drug-supplemented media. Only 12 (4.9%) serows were shown to have drug-resistant E. coli. No transferable R plasmid was detected among a total of 87 resistant strains from wild serows. In contrast, all 33 captive serows except one which was kept only one day after capture, showed resistant E. coli and 20 (60.6%) serows were excreting R plasmid-carrying E. coli. Of 161 drug-resistant strains from captive serows, 50 (31.1%) were found to carry R plasmids. Wild serows seemed to readily change to harbor resistant E. coli almost as soon they were reared in human areas without direct exposure to drugs. These results lead to the conclusion that drug-resistant E. coli can probably be used as microbial indicator for natural environmental pollution.