In this study, previous use of penicillin or ampicillin and private medical coverage were associated with having DRSP. Children with nonmeningeal invasive disease responded equally well to penicillin regardless of the penicillin susceptibility of their pneumococcal isolate.

The Pan American Health Organization (PAHO) has conducted a study of Streptococcus pneumoniae in six Latin-American countries: Argentina, Brazil, Chile, Colombia, Mexico, and Uruguay. Sterile site isolates from children aged < or =5 years showing clinical symptoms of pneumonia (as defined by the clinical criteria of WHO), meningitis, sepsis or bacteremia (without infectious foci), arthritis, and peritonitis were the source of most of the invasive pneumococcal isolates collected between the end of 1993 and 1996 in the six participating countries. Partial characterization of these isolates (antibiotic resistance and serotyping) have already been described (Microbial Drug Resistance 3:(2):131-163, 1997). In the next phase of the study, 326 S. pneumoniae isolates with reduced penicillin susceptibility were transferred to the Laboratory of Microbiology at The Rockefeller University for molecular characterization, and a summary and overview of the findings is described in this article. Some of the most interesting findings were as follows: (1) There was a surprisingly high representation of two internationally spread clones, which made up >80% of the strains with penicillin MIC of 1 microg/ml or higher; most of these isolates were recovered in large cities, supporting the likelihood that the source of these clones is through international travel. (2) The frequency of resistance to trimethoprim/sulfamethoxazole was extremely high (present in 85% of all isolates with decreased penicillin susceptibility). (3) None of these isolates was resistant to ofloxacin, and macrolide resistance was rare (present in 6.4% of the isolates). (4) There was an apparent inverse relationship between level of penicillin resistance and genetic diversity. (5) There were striking differences in the "microbiologic profiles" of the six different Latin-American countries.

Four hundred ninety-nine methicillin-resistant

Since 1992 there have been seven major outbreaks of cholera in Argentina. Susceptibility analysis of 1,947 isolates (40% of reported cases) of Vibrio cholerae O1 biotype El Tor suggested the presence of extendedspectrum ␤-lactamases (ESBLs) in 28 isolates. Because of their different susceptibility profiles, V. cholerae isolates M1502, M1516, M1573, and M3030 (all of which are of the Ogawa serotype) were selected for the present study. By susceptibility analysis, isoelectric focusing, and PCR-based restriction fragment length polymorphism analysis, CTX-M-type enzymes were identified in three isolates, whereas a PER-2-type enzyme, in addition to a TEM-1-like enzyme, was identified in the other isolate. The presence of these ESBLs in V. cholerae isolates resulted in MICs well below those commonly observed for members of the family Enterobacteriaceae. Genes that encode both ESBLs were transferred to Escherichia coli by conjugation, together with all determinants of resistance to non-␤-lactam antibiotics (gentamicin, kanamycin, and sulfamethoxazole for all isolates; amikacin and streptomycin for three isolates; trimethoprim, tetracycline, and chloramphenicol for two isolates). Plasmid profile analysis and Southern blotting revealed the presence of single plasmids of about 150 kb in the four V. cholerae isolates and their respective transconjugants and revealed that the plasmids harbored genes encoding CTX-M-type or PER-2-type ESBLs. These results strongly suggest the broad spread of these ESBLs among genera belong to families other than the Enterobacteriaceae.

One hundred forty-eight recent methicillin-resistant Staphylococcus aureus (MRSA) isolates collected from 13 hospitals in Argentina were examined for antibiotic susceptibility and clonal type, using hybridization with DNA probes specific for mecA and Tn554, and pulsed-field gel electrophoresis (PFGE) of chromosomal SmaI digests. The majority of the isolates (62.2%) shared the common PFGE B pattern and carried variants of mecA and Tn554 polymorphs characteristic of an MRSA clone widely spread in Brazilian hospitals. Similarly to the Brazilian isolates, the MRSA clone recovered in the Argentinian hospitals (XI::B::B) and its close relatives (XI::B'::B, XI::AA::B, XI::M::B, XI::omega omega::B, and III::W::B) showed susceptibility to spectinomycin and resistance to numerous antibacterial agents, including beta-lactams, tetracycline, aminoglycosides, macrolides, trimethoprim/sulfamethoxazole, ciprofloxacin, and fosfomycin, and more than 60% of the isolates were also resistant to chloramphenicol and rifampin. The XI::B::B MRSA clone represented the majority of isolates recovered in most of the hospitals, nine of which were located in the city of Buenos Aires, three in the province of Buenos Aires, and one in the province of Tucumán, 1,312 km northwest of the city of Buenos Aires. The observations document further geographic expansion of this South American MRSA clone across national boundaries.

In a previous study, an analysis of 77 ampicillin-nonsusceptible (resistant plus intermediate categories) strains of Vibrio cholerae non-O1, non-O139, isolated from aquatic environment and diarrheal stool, showed that all of them produced a ␤-lactamase with a pI of 5.4. Hybridization or amplification by PCR with a probe for bla TEM or primers for bla CARB gene families was negative. In this work, an environmental ampicillinresistant strain from this sample, ME11762, isolated from a waterway in the west region of Argentina, was studied. The nucleotide sequence of the structural gene of the ␤-lactamase was determined by bidirectional sequencing of a Sau3AI fragment belonging to this isolate. The gene encodes a new 288-amino-acid protein, designated CARB-7, that shares 88.5% homology with the CARB-6 enzyme; an overall 83.2% homology with PSE-4, PSE-1, CARB-3, and the Proteus mirabilis N29 enzymes; and 79% homology with CARB-4 enzyme. The gene for this ␤-lactamase could not be transferred to Escherichia coli by conjugation. The nucleotide sequence of the flanking regions of the bla CARB-7 gene showed the occurrence of three 123-bp V. cholerae repeated sequences, all of which were found outside the predicted open reading frame. The upstream fragment of the bla CARB-7 gene shared 93% identity with a locus situated inside V. cholerae's chromosome 2. These results strongly suggest the chromosomal location of the bla CARB-7 gene, making this the first communication of a ␤-lactamase gene located on the VCR island of the V. cholerae genome.

beta-Lactamases with pIs of 5.4 and 8.1 were detected by hydrolysis of ampicillin and ceftriaxone respectively among cefotaxime resistant serovars of Salmonella spp. suggesting TEM-1 and an unidentified extended spectrum beta-lactamase have spread throughout these strains in Argentina.

Streptococcus pneumoniae (SPN) is the most common cause of invasive infections in children, with high levels of mortality in developing countries. An increase in frequency of penicillin-resistant strains is reported in most parts of the world. A study was undertaken in Argentina and 5 other countries of the region, to determine the type distribution and penicillin resistance rate of SPN isolated from invasive infections in children less than 5 years old. Between June 1994 and March 1996, a total of 505 SPN isolated from sterile sites were collected from 15 hospitals located in 9 cities of different geographic areas. Clinical and epidemiological data from 443 children were analyzed. Sixty five percent SPN were isolated from children less than 2 years old. Pneumonia was the clinical diagnosis in 58% of the cases, meningitis in 22%, and sepsis in 10.6%. Isolates were recovered from blood (51.2%), pleural fluid (22.7%), CSF (20.7%), and other sterile sites (5.4%). Thirty different pneumococcal capsular types were identified and the 10 most frequent in descending order were: 14, 5, 1, 6A/6B, 7F, 9V, 19F, 19A, 16F y 23F, representing 89.3% of the total. Overall, 13.1% of isolates showed intermediate resistance to penicillin while 11.3% showed high resistance. Lethality was 8.8%, without correlation with penicillin-resistance and/or type. These result should be used in selecting the optimal combination of specific types for a conjugate vaccine, useful in children less than 2 years old and for considering therapeutic strategies for invasive pneumococcal infections.