Dissemination of Salmonella enterica serotype agona and multidrug-resistant Salmonella enterica serotype typhimurium in Cuba.

The molecular epidemiology, antimicrobial susceptibility, and mechanisms of resistance of 34 Salmonella spp. strains causing acute gastroenteritis, isolated from different provinces in Cuba, were determined. Sixty-four percent of the strains showed multiresistance. Salmonella typhimurium was the most frequent with 15 strains (44%), 13 of which belonged to phagotype 104 and presented similar genetic profiles of pulsed field gel electrophoresis. High levels of resistance to tetracycline (53%), spectinomycin (50%), ampicillin (44%), and chloramphenicol (41%) were found. Resistance to tetracycline was associated with the tet G and tet A genes. Resistance to ampicillin was caused by the presence of beta-lactamases, mainly the CARB type. The floR gene was the main mechanism of resistance to chloramphenicol. Our results showed an antimicrobial susceptible clone of Salmonella enterica serotype Agona in two separate regions. This is the first report of the widespread dissemination of a multiresistant clone of S. enterica serotype Typhimurium definitive phage type 104 in Cuba.

[Serotype and phage type distribution of human Salmonella strains isolated in Spain, 1997-2001]. / Distribución de los serotipos y fagotipos de Salmonella de origen humano aislados en España en 1997-2001.

INTRODUCTION: Salmonellosis is one of the most frequent causes of gastroenteritis in Spain. Serotyping is the gold standard epidemiological marker for subdividing Salmonella spp. strains. A small number of serotypes are very frequently isolated, reducing the discriminatory power of serotyping. Thus, to increase our knowledge of Salmonella spp. epidemiology, additional epidemiological markers, such as phage typing, should be used for this purpose. METHODS: Salmonella spp. strains of human origin sent to the Laboratorio Nacional de Referencia de Salmonella y Shigella (LNRSSE, Spanish Reference Laboratory for Salmonella and Shigella) between 1997 and 2001 were serotyped using conventional agglutination methods, and Enteritidis, Typhimurium, Hadar, Virchow and Typhi serotypes were additionally phage typed according to internationally-developed schemes. RESULTS: A total of 30,856 Salmonella spp. strains, isolated in the majority of Spanish Autonomous Communities, were analyzed. Enteritidis (51%) and Typhimurium (24%) were the most frequently isolated serotypes. The following were the most frequent serotype/phage type combinations: Enteritidis/PT1 (18%), Enteritidis/PT4 (15%), Enteritidis/PT6a (5%), Typhimurium/DT104 (5%) and Enteritidis/PT6 (3%). The serotype Enteritidis/PT1 showed the greatest increase over the period studied, from 11.61% in 1997 to 24.74% in 2001. CONCLUSIONS: A hierarchical typing approach for Salmonella spp., using serotyping coupled with phage typing allowed a higher level of discrimination among Salmonella serotypes. Application of this approach in epidemiological studies could be highly useful for early characterization of related strains.

Distribución de los serotipos y fagotipos de Salmonella de origen humano aislados en España en 1997-2001 / Serotype and phage type distribution of human Salmonella strains isolated in Spain, 1997-2001

INTRODUCCIÓN. La salmonelosis continúa siendo una de las causas principales de gastroenteritis en España, siendo la serotipificación el marcador epidemiológico universalmente utilizado para la caracterización de los aislamientos de Salmonella spp. Algunos serotipos se identifican muy frecuentemente, reduciendo el poder de discriminación de esta técnica. Por ello, para el estudio epidemiológico de las salmonelosis producidas por estos serotipos es necesario utilizar marcadores complementarios como la fagotipificación. MÉTODOS. Se serotipificaron, por aglutinación directa, las cepas de Salmonella spp. de origen humano recibidas en el Laboratorio Nacional de Referencia de Salmonella y Shigella (LNRSSE) entre los años 1997 y 2001 y se fagotipificaron, según esquemas internacionales, las cepas de los serotipos Enteritidis, Typhimurium, Hadar,Virchow y Typhi. RESULTADOS. Se analizaron 30.856 cepas de Salmonella spp. procedentes de la mayoría de las Comunidades Autónomas. Los serotipos Enteritidis (51%) y Typhimurium (24%) fueron los mayoritarios. Las combinaciones serotipo/fagotipo más frecuentes fueron: Enteritidis/FT1 (18%), Enteritidis/FT4 (15%),Enteritidis/FT6a (5%), Typhimurium/FT104 (5%)y Enteritidis/FT6 (3%). Las cepas del serotipo Enteritidis/FT1 tuvieron el mayor aumento en este período de tiempo, pasando del 11,61% en 1997al 24,74% en 2001. CONCLUSIONES. La utilización jerárquica de la serotipificación y posteriormente de la fagotipificación en cepas de Salmonella spp. de los serotipos más frecuentes aumentó enormemente el poder de discriminación de la serotipificación. Su aplicación en estudios epidemiológicoses de gran utilidad en la caracterización temprana de cepas relacionadas (AU)

INTRODUCTION. Salmonellosis is one of the most frequent causes of gastroenteritis in Spain. Serotyping is the gold standard epidemiological marker for subdividing Salmonella spp. strains. A small number of serotypes are very frequently isolated, reducing the discriminatory power of serotyping. Thus, to increase our knowledge of Salmonella spp. epidemiology, additional epidemiological markers, such as phage typing, should be used for this purpose. METHODS. Salmonella spp. strains of human origin sent to the Laboratorio Nacional de Referencia de Salmonellay Shigella (LNRSSE, Spanish Reference Laboratory for Salmonella and Shigella) between 1997 and 2001 were serotyped using conventional agglutination methods, and Enteritidis, Typhimurium, Hadar, Virchow and Typhiserotypes were additionally phage typed according to internationally-developed schemes. RESULTS. A total of 30,856 Salmonella spp. strains, isolatedin the majority of Spanish Autonomous Communities,were analyzed. Enteritidis (51%) and Typhimurium (24%)were the most frequently isolated serotypes. The following were the most frequent serotype/phage type combinations: Enteritidis/PT1 (18%), Enteritidis/PT4 (15%),Enteritidis/PT6a (5%), Typhimurium/DT104 (5%) and Enteritidis/PT6 (3%). The serotype Enteritidis/PT1 showed the greatest increase over the period studied, from 11.61%in 1997 to 24.74% in 2001. CONCLUSIONS. A hierarchical typing approach for Salmonellaspp., using serotyping coupled with phage typing allowed a higher level of discrimination among Salmonella serotypes. Application of this approach in epidemiological studies could be highly useful for early characterization of related strains (AU)

Mechanism of resistance to several antimicrobial agents in Salmonella Clinical isolates causing traveler's diarrhea.

The evolution of antimicrobial resistance in Salmonella isolates causing traveler's diarrhea (TD) and their mechanisms of resistance to several antimicrobial agents were analyzed. From 1995 to 2002, a total of 62 Salmonella strains were isolated from stools of patients with TD. The antimicrobial susceptibility to 12 antibiotics was determined, and the molecular mechanisms of resistance to several of them were detected as well. The highest levels of resistance were found against tetracycline and ampicillin (21 and 19%, respectively), followed by resistance to nalidixic acid (16%), which was mainly detected from 2000 onward. Molecular mechanisms of resistance were analyzed in 16 isolates. In these isolates, which were resistant to ampicillin, two genes encoding beta-lactamases were detected: oxa-1 (one isolate) and tem-like (seven isolates [in one strain concomitantly with a carb-2]). Resistance to tetracycline was mainly related to tetA (five cases) and to tetB and tetG (one case each). Resistance to chloramphenicol was related to the presence of the floR and cmlA genes and to chloramphenicol acetyltransferase activity in one case each. Different genes encoding dihydrofolate-reductases (dfrA1, dfrA12, dfrA14, and dfrA17) were detected in trimethoprim-resistant isolates. Resistance to nalidixic acid was related to the presence of mutations in the amino acid codons 83 or 87 of the gyrA gene. Further surveillance of the Salmonella spp. causing TD is needed to detect trends in their resistance to antimicrobial agents, as we have shown in our study with nalidixic acid. Moreover, such studies will lead to better treatment and strategies to prevent and limit their spread.

Detection of integrons and antibiotic-resistance genes in Salmonella enterica serovar Typhimurium isolates with resistance to ampicillin and variable susceptibility to amoxicillin-clavulanate.

We characterized 29 antimicrobial-resistant Salmonella enterica serovar Typhimurium strains, including four belonging to the monophasic variant 4,5,12:i:-, mostly isolated from infants. They were selected from 3230 strains isolated in the years 1990-2001 on the basis of resistance to ampicillin and variable susceptibility to the amoxicillin-clavulanate combination. Twenty-three strains were resistant to more than four antibiotics. All the strains carried the bla(TEM) gene and most were able to transfer this gene by conjugation. Sequencing of the gene from one of the amoxicillin-clavulanate-resistant strains allowed identification of the encoded beta-lactamase as TEM-1; all of these strains carried a second gene encoding beta-lactamase production, either pse-1 or oxa1. However, the association of bla(TEM) plus pse-1 genes did not always confer resistance to amoxicillin-clavulanate. The pse-1 gene, found in 17 strains, was located in the Salmonella Genomic Island-1 (SGI1), which carries two integrons and encodes multiple drug-resistance. None of the oxa1-bearing strains had the SGI1, yet this gene was found as part of an integron that also carried the aadA1 gene and was not plasmid-associated. Thirteen of the strains harbouring SGI1 belonged to the definitive phage type (DT) 104, and most of those remaining to DT104b and U302; particularly, strains carrying the oxa1-aadA1 integron belonged to the last two phage types. Pulsed field electrophoresis confirmed the clonal organization of DT104 strains, whereas U302 strains fell into different groups, depending on their resistance determinants.