José Moacir Marin

Detection of streptococcus mutans and streptococcus sobrinus in dental plaque samples from Brazilian preschool children by polymerase chain reaction

The purposes of this study were to detect S. mutans and S. sobrinus by polymerase chain reaction (PCR) amplification, and to relate their presence to the incidence of dental caries in 42 Brazilian preschool children. Dental plaque samples were collected from the cervical margin of all erupted teeth of 5-6 years old children with primary dentition, using a sterile explorer. Examination of the dmft (decayed, missing, filled teeth) index, performed following the World Health Organization (WHO) caries diagnostic criteria, showed a 2.71 score. Prevalence of S. mutans and S. sobrinus was respectively, of 85.7% and 14.3%; no dental plaque sample was either positive or negative for both bacterial species. Children harboring either S. mutans or S. sobrinus presented the same caries prevalence. PCR showed good discriminative ability for differentiation between these species, and suggested that it is a technique suitable for epidemiological studies on mutans streptococci.

Occurrence of toxigenic Escherichia coli in raw milk cheese in Brazil

The occurrence of toxigenic Escherichia coli in raw milk cheese was surveyed in Middle Western Brazil. Fifty samples of cheese from different supermarkets were analyzed for E.coli. The isolates were serotyped and screened for the presence of verotoxigenic E. coli (VTEC) and enterotoxigenic E. coli (ETEC) by Polymerase Chain Reaction (PCR). The susceptibility to thirteen antimicrobial agents was evaluated by the disk diffusion method. E.coli were recovered from 48 (96.0%) of the samples. The serogroups identified were O125 (6.0%), O111 (4.0%), O55 (2.0%) and O119 (2.0%). Three (6.0%) and 1(2.0%) of the E.coli isolates were VTEC and ETEC, respectively. Most frequent resistance was observed to the following antimicrobials: cephalothin (60.0%), nalidixic acid (40.0%), doxycyclin (33.0%), tetracycline (31.0%) and ampicillin (29.0%).

Virulence factors of uropathogenic Escherichia coli from a University Hospital in Ribeirão Preto, São Paulo, Brazil

The aim of the study was to determine the occurrence of virulence genes expressing fimbriae, production of hemolysin, colicin and aerobactin among a hundred Escherichia coli isolates obtained from in-and outpatients of a tertiary-care teaching hospital, between July and August 2000, showing clinical and laboratory signs of urinary tract infection (UTI). The presence of genes (pap, afa, sfa) for fimbriae expression was assayed using specific primers in a polymerase chain reaction. Among the isolates studied, the prevalence of the virulence factors was 96.0%, 76.0%, 24.0%, for hemolysin, aerobactin and colicin, respectively; the prevalence of genes coding for fimbrial adhesive systems was 32.0%, 19.0% and 11.0% for pap, sfa and afa respectively. The strains isolated from the outpatients displayed a greater number of virulence factors compared to those from hospitalized subjects, emphasizing the difference between these two kinds of patients.

Virulence factors of Escherichia coli isolated from diarrheic calves

Cento e setenta e tres cepas de Escherichia coli isoladas de bezerros com diarreia provenientes da regiao noroeste do estado de Sao Paulo foram examinadas para a producao de enterotoxinas termolabil (LT) e termoestavel (ST), e examinadas quanto a presenca de fatores de virulencia associados a colibacilose bovina. Oitenta e cinco (49,1%) das cepas de E. coli produziram toxinas, 53 cepas foram detectadas como produtoras de toxina STa, e nove dessas cepas tambem produziam toxina LT. Foram identificadas pela reacao em cadeia de polimerase 23 cepas portadoras do gene LT-II. Nove (5,2%) das cepas apresentavam os genes de toxina Shiga: quatro o gene stx 2, quatro o gene stx 1 e uma cepa apresentava os dois genes. Tres das cepas que apresentavam o gene stx1 tambem possuiam o gene eae. Entre as cepas de E. coli examinadas quanto a susceptibilidade a 10 agentes antimicrobianos, a resistencia a cefalotina (46,1%) foi a mais comumente observada, seguida pelas resistencias a tetraciclina (45,7%), trimetropima-sulfadiazina (43,3%) e ampicilina (41,0%). Todas as cepas isoladas apresentaram resistencia a pelo menos dois antimicrobianos, sendo a multirresistencia detectada em elevada frequencia. Algumas toxinas e fatores de virulencia, produzidos por essas cepas de E. coli podem estar envolvidos em doencas humanas. O alto nivel de resistencia a agentes antimicrobianos, apresentado pelas cepas isoladas, constitue motivo de preocupacao em saude publica.

Multidrug-resistant urinary tract isolates of escherichia coli from Ribeirão Preto, São Paulo, Brazil

Multiple resistances to antimicrobial drugs arising in Escherichia coli isolates may complicate therapeutic management of urinary tract infection (UTI) by this organism. In order to assess the multidrug resistance (MDR) among urinary E. coli isolates, we have tested 11 antimicrobial drugs against 67 isolates from outpatients attended in a tertiary-care teaching hospital and of 78 isolates from a municipal health unit, respectively in Ribeirão Preto, State of São Paulo, Brazil. Seventy-six percent and 22% of the isolates from the tertiary-care hospital and the municipal unit, respectively, were resistant to three or more different classes of agents, and were considered to present MDR. Among the isolates from the hospital patients, 73.0%, 65.0%, 58.0%, 58.0% and 31.0% were resistant to tetracycline, ampicillin, cephalothin, trimethoprim-sulfamethoxazole (TMP/SMX) and norfloxacin, respectively; resistance from the municipal unit patients were 31.0%, 37.0%, 8.0%, 29.0% and 12.0% respectively, to the same drugs. The predominant phenotype among the MDR isolates presented is ampicillin, TMP/SMX and tetracycline resistance. The high prevalence of drug resistance among UTI patients calls for continuous surveillance to assure effective control of this infection.

Isolation of Shiga toxigenic Escherichia coli from butcheries in Taquaritinga city, State of São Paulo, Brazil

Shiga toxigenic Escherichia coli (STEC) has been implicated as the cause of several human diseases. Samples (ground beef, grinding-machines and human hands) from 23 butcheries were assayed for E. coli using standard microbiological methods, and 287 isolates were submitted to polymerase chain reaction for the detection of stx 1, stx 2 and eae genes. Four STEC isolates were recovered, two from ground beef and two from grinding-machines; all harbored the stx 2 gene and were negative for the eae gene. All E. coli isolates including the four STEC were screened for antibiotic resistance. High levels of resistance against several antimicrobial agents were detected; those most commonly observed were to tetracycline (76.6%), amoxicillin (64.1%) and cephalothin (58.8%). Such high levels of antimicrobial resistance highlight the need for a more rational use of these agents in cattle.

An epidemiological study of Haemophilus influenzae at a Brazilian day care center

Day care centers are a relatively new phenomenon in Brazil that bring together large numbers of young children susceptible to contagious diseases. Haemophilus influenzae (Hi) is an important infection in the age range of those attending day care centers. In the present study, the carriage rate of Haemophilus influenzae was identified in 38 day care attendees age 6 to 37 months, and 23 staff members, at a day care center in Ribeirão Preto-São Paulo, in 1997. To identify the carriers, two nasopharyngeal swabs were collected; one in July and one in December. The rate of H. influenzae carriers among the children was 77%. Only 2 of 23 staff members (9%) had Hi. Among the children, there were 58 isolates in the two sampling periods; 6 of the Hi were serotype b, 1 was serotype e, and 48 isolates were non-typeable. Two were identified as H. parainfluenzae. One adult had a non-typeable Hi and 1 had H. paraphrohaemolyticus. Three of the 6 children with type B had received a conjugate vaccine against H. influenzae type b, but they still carried this bacterium in the nasopharynx (50%). Forty ribotype patterns were found among the isolates, showing a high exchange rate of nontypeable H. influenzae carriers. The results indicate that, because of the high and changing biotype of Hi carriage, day care centers should be carefully monitored as potential point source of HI disease in the community.

Beef Carcass Contamination by Shiga Toxin-Producing Escherichia coli Strains in an Abattoir in Brazil: Characterization and Resistance to Antimicrobial Drugs

A survey was performed to estimate the frequency of Escherichia coli and Shiga toxin-producing E. coli (STEC) in carcasses obtained from an abattoir in Brazil between February 2006 and June 2007. A total of 216 beef carcasses were sampled at three stages of the slaughter process--preevisceration, postevisceration, and postprocessing--during the rain and dry seasons, respectively. Of the carcasses sampled, 58% were preevisceration E. coli positive, 38% were postevisceration positive, and 32% postprocessing positive. At the postprocessing stage, the isolation of E. coli was twice as high in the rain season. E. coli was isolated from 85 carcasses of which only 3 (1.4%) were positive for stx-encoding genes. No E. coli O157 serogroup isolates were detected. No antimicrobial resistance was found in nine of the isolates (10% of the total). The most frequent resistances were seen against cephalothin (78%), streptomycin (38%), nalidixic acid (36%), and tetracycline (30%). Multidrug resistance (MDR) to three or more antimicrobial agents was determined in 28 (33%) E. coli isolates. The presence of STEC and MDR strains among the isolates in the beef carcasses emphasizes the importance of proper handling to prevent carcass contamination.

Isolation of Pseudomonas aeruginosa strains from dental office environments and units in Barretos, state of São Paulo, Brazil, and analysis of their susceptibility to antimicrobial drugs

A wide variety of opportunistic pathogens has been detected in the tubing supplying water to odontological equipment, in special in the biofilm lining of these tubes. Among these pathogens, Pseudomonas aeruginosa, one of the leading causes of nosocomial infections, is frequently found in water lines supplying dental units. In the present work, 160 samples of water, and 200 fomite samples from forty dental units were collected in the city of Barretos, State of Sao Paulo, Brazil and evaluated between January and July, 2005. Seventy-six P. aeruginosa strains, isolated from the dental environment (5 strains) and water system (71 strains), were tested for susceptibility to six antimicrobial drugs most frequently used against P. aeruginosa infections. Susceptibility to ciprofloxacin, followed by meropenem was the predominant profile. The need for effective means of reducing the microbial burden within dental unit water lines is emphasized, and the risk of exposure and cross-infection in dental practice, in special when caused by opportunistic pathogens like P. aeruginosa, are highlighted.

Prevalence of mutans streptococci in 93 members from six Brazilian families

Several studies report that mutans streptococci (MS) are closely associated with caries in humans and that there is a correlation between the number of carious lesions and the levels of MS in the saliva of children and adults. The presence of MS in the saliva of 93 members of six Brazilian families with at least 3 generations was investigated. Samples of whole unstimulated saliva were collected and diluted. Aliquots of 50 microliters of each suspension were dropped onto SB20 agar and incubated in a candle jar at 37 degrees C for 72 h. Colonies resembling MS were counted, collected, seeded in thioglycoilate medium and subjected to biochemical typing. Mutans streptococci were isolated from 80 subjects (86.0%) and the counts ranged from 3.0 x 10(2) (log 2.477) to 1.6 x 10(8) (log 8.204) CFU/ml of saliva. All of the 73 adults were colonized by MS, but the bacteria were detected in only 7 (35.0%) of the 20 children evaluated. Streptococcus mutans occurred in 78 subjects (97.5%), and 51 (63.7%) were monocolonized. S. sobrinus occurred in 29 individuals (36.3%) and 2 (2.5%) were monocolonized. Twenty-seven (33.8%) subjects were multicolonized with S. mutans and S. sobrinus. This study showed a high prevalence (86.0%) of mutans streptococci in the saliva of members of the studied families, which suggests the risk of intrafamilial transmission.