Evaluation of PCR in the diagnosis of canine leishmaniasis in two different epidemiological regions: Campinas (SP) and Teresina (PI), Brazil.

SUMMARY Using the polymerase chain reaction (PCR) test for diagnosis of canine leishmaniasis has greater sensitivity and specificity than culture and visualization of the parasite. This study compares PCR for the diagnosis of the genus and species of Leishmania with serological techniques used for the control of canine visceral leishmaniasis (CVL) in Brazil, considering two regions. We analysed peripheral blood samples collected from 195 dogs in the Campinas (SP) and Teresina (PI) regions. ELISA was performed as a serological method and PCR was performed using specific primers for the genus Leishmania spp. and the species Leishmania chagasi. In Campinas, a greater sensitivity of PCR (88.24%) (P = 0.0455) compared to Teresina (14.71%) (P < 0.0001) was observed, and an agreement was observed for Cohen's kappa index (0.9096). Both PCR and ELISA showed discordance for sensitivity (Campinas 100%, Teresina 21.74%), specificity (Campinas 30.77%, Teresina 100%), positive predictive value (Campinas 68.97%, Teresina 100%), negative predictive value (Campinas 100%, Teresina 37.94%) and Cohen's kappa index (0.1238). This study confirms the importance of PCR in analysis of the canine reservoir, and as an effective method for the detection of active and recent infection.

Controlling a vancomycin-resistant enterococci outbreak in a Brazilian teaching hospital.

The purpose of this article was to describe a 2.5-year interventional program designed to control the dissemination after a large hospital outbreak of vancomycin-resistant enterococci (VRE) in a tertiary-care university hospital. A VRE working group was designated to work specifically on controlling VRE intrahospital dissemination after the detection of the first VRE infection at in our hospital in June 2007. The intervention consisted in the interruption of new admissions during a period of 15 days and closure of the index case unit, microbiological surveillance of rectal swabs for VRE, cohorting patients and staff, immediate application of contact precautions, and continuous education. From July 2007 to December 2009, 8,692 rectal swabs were cultured for VRE and 321 (3.7%) were positive. An expressive reduction of the detection of new positive rectal swabs cultures was seen during the year 2009 (1.5%) when compared to 2008 (4.2%) and 2007 (7.2%) (p < 0.005). The annual ratio of VRE per 1,000 admissions reduced from 20.3 in 2007 to 10.07 and 3.82 in 2008 and 2009, respectively (p < 0.001). The continuous microbiologic surveillance for VRE and strict and prompt contact precautions for VRE patients were the fundamental aids in the control of VRE.

Evaluation of antimicrobial effectiveness of C-8 xylitol monoester as an alternative preservative for cosmetic products.

Xylitol is a natural sugar derived from plants, fruits and vegetables, whose antimicrobial properties are described in the literature. This study aimed to evaluate the antimicrobial effectiveness of C-8 xylitol monoester, for its use as a preservative in cosmetic formulations. The minimum inhibitory concentration (MIC) was determined by the broth macrodilution method, and the antimicrobial effectiveness of C-8 xylitol monoester was determined by using challenge test method. The results obtained in the determination of minimum inhibitory concentration are between 1.0% and 1.25% for Staphylococcus aureus, Escherichia coli and Candida albicans and between 1.0% and 1.5% for Pseudomonas aeruginosa and Aspergillus niger. The amount of 1% of C-8 xylitol monoester was added to the lotion used in the challenge test, observing a rapid decline in the number of CFU g(-1) in stages of evaluation after contamination of the product by all bacteria. The same occurs in relation to C. albicans, which shows a 90% reduction in the number of CFU g(-1). Regarding A. niger, similar reduction is observed when pH value of the lotion is adjusted from 5.5 to 7.0. The results indicate that under the tests conditions, C-8 xylitol monoester has antimicrobial activity and could be considered as an alternative preservative for cosmetic formulations.

Challenges in the identification of Chryseobacterium indologenes and Elizabethkingia meningoseptica in cases of nosocomial infections and patients with cystic fibrosis.

Rare nonfermenting Gram-negative bacilli, such as Chryseobacterium indologenes and Elizabethkingia meningoseptica, have clinical importance in nosocomial infections and cystic fibrosis (CF), and their identification is a challenge to microbiology laboratories. Thus, the objective of this study was to verify the performance of phenotypic and mass spectrometry (matrix-assisted desorption ionization-time of flight mass spectrometry, MALDI-TOF MS) methods to identify C. indologenes and E. meningoseptica. In this context, the results obtained with phenotypic methods-namely manual biochemical and automated VITEK 2 (bioMérieux, Marcy l'Etoile, France) and Phoenix tests (Becton Dickinson (BD), San Diego, CA, USA)-and by MALDI-TOF MS-namely MALDI-TOF VITEK MS (MALDI-MS; bioMérieux) and MALDI-TOF BioTyper (MALDI-BD; BD)-of 22 isolates (blood cultures of patients with nosocomial infection (n = 15) and from patients with CF (n = 7)), initially identified as C. indologenes and E. meningoseptica, were compared. As result, using the manual phenotypic method, it was possible to identify the species level in 18/22; no identification was found in 4/22. There was a low agreement level between manual and VITEK 2 automated phenotypic methods when considering the genus level. The greatest agreement for genus-level identification occurred in MALDI-TOF MS equipment (15/22). When comparing all methods to identify the 22 isolates, there was agreement of 4/22 at the genus level and of 4/22 at the species level. In conclusion, there is low agreement level among identification methods of C. indologenes and E. meningoseptica. Although MALDI-TOF MS equipment shows a higher agreement level among them, results present low levels of confidence.

Isolation in Brazil of nosocomial Staphylococcus aureus with reduced susceptibility to vancomycin.

OBJECTIVE: To evaluate the possible presence of vancomycin-resistant Staphylococcus aureus (VRSA) in a Brazilian hospital. DESIGN: Epidemiological and laboratory investigation of nosocomial VRSA. METHODS: 140 methicillin-resistant S aureus strains isolated between November 1998 and October 1999 were screened for susceptibility to vancomycin. The screening was carried out by using brain-heart infusion agar (BHIA) supplemented with 4, 6, and 8 microg/mL of vancomycin. The minimum inhibitory concentration (MIC) determination was carried out as standardized by the National Committee for Clinical Laboratory Standards using the broth macrodilution, agar-plate dilution, and E-test methods. PATIENTS: Hospitalized patients exposed to vancomycin. RESULTS: 5 of the 140 isolates had a vancomycin MIC of 8 microg/mL by broth macrodilution, agar plate dilution, and E-test methods. Four VRSA strains were isolated from patients in a burn unit who had been treated with vancomycin for more than 30 days, and one from an orthopedic unit patient who had received vancomycin treatment for 7 days. Pulsed-field gel electrophoresis characterized four of the VRSA strains as belonging to the Brazilian endemic clone. All five strains were negative for vanA, vanB, and vanC genes by polymerase chain reaction. Transmission electron microscopy of the five strains revealed significantly thickened cell walls. One patient died due to infection caused by the VRSA strain. CONCLUSIONS: This is the first report of isolation of VRSA in Brazil and the first report of isolation of multiple VRSA strains from one facility over a relatively short period of time. This alerts us to the possibility that VRSA may be capable of nosocomial transfer if adequate hospital infection control measures are not taken.

Nosocomial infections caused by multiresistant Pseudomonas aeruginosa.

A case-control study was done to evaluate factors associated with nosocomial infections by multiresistant Pseudomonas aeruginosa (MRPA). Results showed that MRPA was associated with the use of immunosuppressive and antimicrobial drugs. Five typing methods indicated that the MRPA infections were due to multiple strains rather than a single strain.

Short report: Decrease in seroprevalence of antibodies to hantavirus in rodents from 1993-1994 hantavirus pulmonary syndrome case sites.

Rodent trapping was conducted at seven hantavirus pulmonary syndrome (HPS) case sites from June 1993 to March 1994 during the HPS outbreak in the southwestern United States. To determine if there were changes in the rodent population or the hantavirus seroprevalence in rodents since the HPS outbreak, rodents were trapped at the same sites three years later using the same trapping protocol. The trap success decreased from the numbers trapped during the outbreak, however, the number of Peromyscus, as a percentage of the total rodents captured, did not noticeably decrease. In addition, the seroprevalence of hantavirus antibodies in Peromyscus decreased significantly (P < 0.0001).

Evaluation of malaria surveillance using retrospective, laboratory-based active case detection in four southwestern states, 1995.

The global resurgence of malaria has raised concerns of the possible reintroduction of indigenous transmission in the United States. The Centers for Disease Control and Prevention's National Malaria Surveillance System, using data supplied by state and local health departments (SLHDs), is maintained to detect local malaria transmission and monitor trends in imported cases. To determine the completeness of reporting of malaria cases to SLHDs, cases identified by local surveillance systems were compared with those identified through active case detection conducted at all laboratories that receive clinical specimens from 11 metropolitan areas in Arizona, California, New Mexico, and Texas. Of the 61 malaria cases identified through either local surveillance or active case detection, 43 (70%) were identified by SLHDs (range by metropolitan area = 50-100%) and 56 (92%) through active case detection. High percentages of cases were identified by SLHDs in New Mexico (80%) and San Diego County (88%), where laboratories are required to send positive blood smears to the SLHD laboratory for confirmation. Completeness of reporting, calculated using the Lincoln-Peterson Capture-Recapture technique, was 69% for SLHD surveillance systems and 89% for laboratory-based active case detection. The high percentage of cases identified by the 11 SLHDs suggests that the National Malaria Surveillance System provides trends that accurately reflect the epidemiology of malaria in the United States. Case identification may be improved by promoting confirmatory testing in SLHD laboratories and incorporating laboratory-based reporting into local surveillance systems.

Patterns of association with host and habitat: antibody reactive with Sin Nombre virus in small mammals in the major biotic communities of the southwestern United States.

The distribution and prevalence of antibody reactive with Sin Nombre virus were determined in mammals in biotic communities of the southwestern United States. Small mammals (n = 3,069) of 69 species were trapped in nine communities from lower Sonoran desert to alpine tundra. Antibody was found in rodents from all communities (overall prevalence = 6.3%); prevalence was lowest at the altitudinal and climatic extremes (0.4% in desert and 2.0% in alpine tundra). Antibody occurred in 11% of 928 deer mice, 20% of 355 brush mice, 23% of 35 western harvest mice, and 12% of 24 Mexican voles. No infected deer mice were found in desert habitat; prevalence varied from 4% in chaparral to 17% in pinyon-juniper. Brush mice were frequently infected in chaparral and montane forest (25%). Seropositivity was higher in males and in heavier animals, suggesting horizontal transmission among adult males. Decreasing prevalence with age among the youngest deer mice suggests that infected dams confer passive immunity to pups.

Clinical relevance and virulence factors of pigmented Serratia marcescens.

Pigmented Serratia marcescens isolated in a Brazilian hospital were studied with respect to frequency of isolation, serotyping, antibiotic resistance and virulence factors. The serotype most frequent was O6:K14 (53%) and all isolates were resistant to ampicillin, cephalothin and tetracycline. The majority of the isolates (92%) were resistant to the action of human serum and all produced cytotoxins on Vero, CHO, HEp-2 and HeLa cells. These isolates were virulent for mice (LD(50)=10(7) bacteria ml(-1)) and showed virulence factors, but were isolated with low frequency (3. 4%) and caused infection in only 31% of cases. Analysis of serotyping, phage typing and chromosomal DNA revealed at least 13 unrelated strains among pigmented S. marcescens. In conclusion, this work describes a low frequency of isolation of pigmented S. marcescens from clinical specimens, indicating that non-pigmented strains are clinically more significant.

Detection of cytotoxic activity on Vero cells in clinical isolates of Serratia marcescens.

Cytotoxin production was studied in 60 Serratia marcescens strains isolated from hospitalized patients. Association of cytotoxic activity with serotype, source of isolation and presence of plasmids was also evaluated. Thirteen of the 60 S. marcescens strains produced a cytotoxic effect on Vero cells. These strains were isolated from distinct clinical sources and classified into seven different serotypes (O1:H7; O4:NM; O10:NT; O19:NM; O6,14:H4; O6,14:NM and O6,14:H1). No relationship was observed between cytotoxic activity and clinical source or serotypes of the strains. Plasmids from five cytotoxin-producing S. marcescens strains were transferred to E. coli K12/711. The transconjugants did not exhibit cytotoxicity, indicating that the cytotoxic effect is not plasmid-mediated among these strains. Although a cytotoxic activity was demonstrated in filtrates of some S. marcescens strains, further studies should be performed to assess the role of this toxin in pathogenesis.

Characterization of the Brazilian endemic clone of methicillin-resistant Staphylococcus aureus (MRSA) from hospitals throughout Brazil.

The objective of this study was to characterize patterns of the Brazilian endemic clone of methicillin-resistant Staphylococcus aureus (MRSA) from hospitals throughout Brazil. We studied 83 MRSA strains isolated from patients hospitalized in 27 public and private hospitals in 19 cities located in 14 Brazilian states from September, 1995, to June, 1997. The MRSA strains were typed using antibiograms, bacteriophage typing and pulsed field gel electrophoresis (PFGE). The analysis of genomic DNA by PFGE showed that 65 isolates presented the same PFGE pattern. This pattern was present in all of the hospitals studied indicating the presence of an endemic MRSA clone widely disseminated throughout Brazilian hospitals (BEC). All isolates belonging to the BEC proved to be resistant to ciprofloxacin, erythromycin, lincomycin, trimethoprim-sulphamethoxazole, and tetracycline. Variable susceptibility to these drugs was found only in isolates belonging to clones other than the BEC. The results show that, among MRSA, the BEC is common in Brazil. The best method for mapping changes in the frequency of this clone among MRSA is pulsed field gel electrophoresis. Use of molecular mapping is an important tool for monitoring the spread of potentially dangerous microbes.

Nocardia infection in renal transplant recipient: diagnostic and therapeutic considerations.

In the present report the authors discuss the diagnostic difficulties, therapeutic measures and the clinical course of Nocardia infection which occurred among renal transplant recipients at the University Hospital of the Faculty of Medicine of Ribeirão Preto, University of São Paulo (UH-FRP), from 1968 to 1991. Among 500 individuals submitted to renal transplant, 9 patients developed Nocardiosis at varying times after transplant (two months to over two years). All the patients had pulmonary involvement and their most common symptoms were fever, cough and pleural pain. Dissemination of the process is common and three patients presented cutaneous abscesses, four CNS involvement and one had pericarditis due to Nocardia. The diagnostic is quite difficult since there is no specific clinical picture, concomitant infections are frequent and the microorganism presents slow growth in culture (ranging from four to forty days, in our experience). In this report, three cases were only diagnosed by necropsy. The treatment of choice is a combination of Sulfamethoxazole and Trimethoprim (SMX-TMP). In the present series, overall mortality was 77% (7 cases) and in five of the patients who died the diagnosis was late. All the patients who had CNS involvement died.

[Cryptococcus meningitis: clinical course, development and histopathologic aspects depending on the predisposing factors]. / Meningite criptocócica: aspectos clínicos, evolutivos e histopatológicos segundo a condição predisponente.

Seventeen consecutive cases of cryptococcal meningitis diagnosed at Hospital das Clínicas of Ribeirão Preto Medical School (São Paulo State Brazil) between 1969 and 1985 were reviewed. For analysis the patients were separated in 3 groups: I. three patients without immunodeficiency; II. six patients with associated disease: cancer (3), diabetes (2) and alcoholism (1); III. Eight renal transplant recipients that developed cryptococcosis after 18 to 67 months of immunosupression with steroids and azathioprine. The median interval between onset of symptoms and diagnosis of infection was greater in Group II (53 days) than in Groups I (25 days) or III (28 days). Neck stiffness, cranial nerve involvement and papilledema were more frequent in Group I than in Group II or III, but fever and focal neurological signs were observed only in patients of two last groups. Cerebrospinal fluid examination showed a mild lymphocytic pleocytosis in most patients, but transplant cases had polymorphonuclear cells more frequently. Late mortality was higher in patients with underlying disease and the prognosis was better for transplant patients that received effective antifungal therapy. Besides cryptococci, autopsy findings in 8 cases revealed granuloma formation in tissues, except in patients of Group II (2 cases). The differences between the groups suggest that clinical characteristics, evolution and postmortem findings of the cryptococcal meningitis are changed according to type of immunodeficiency presented by the patient.

Conductive hearing loss in individuals with fibrodysplasia ossificans progressiva.

Fibrodysplasia ossificans progressiva (FOP) is a very rare genetic disorder that is characterized by progressive heterotopic ossification of soft tissues and congenital malformation of the great toes. Although previous case studies have reported hearing loss in individuals with FOP, there have been no large-scale studies regarding the nature or cause of the hearing loss. Here, we report the findings of a two-part study. In Part I, we report the findings of a postal survey regarding hearing loss that was sent to 102 individuals with FOP. In Part II, we report the findings of on-site hearing evaluations of eight individuals with FOP. The findings of both studies indicate that individuals with FOP are at risk for hearing loss and that the type of loss is predominantly conductive in nature, similar to that seen in individuals who have otosclerosis.

[Meningoencephalitis due to Pasteurella multocida: clinico-laboratory study of a case in an infant]. / Meningencefalite por Pasteurella multocida: estudo clínico-laboratorial de um caso em lactente.

The authors report a case of Pasteurella multocida meningoencephalitis in a 5 week-old female infant, with special attention to clinical, laboratory and evolutive features. A moderate neurological sequel was observed at follow-up examinations. A brief review of the importance of P. multocida in human pathology is presented on the basis of the international literature, since the authors did not find any Brazilian reports. The most important feature on P. multocida is the prevalence of bacterial meningitis at the extremes of age. Otherwise, significant mistaken was found between Gram stained smears of body fluids for P. multocida and Haemophilus influenzae or Neisseria meningitidis. Because its role in infections following animal bite or scratch and its opportunistic feature, P. multocida must be included among the possible etiologic agent of bacteremia or sepsis in patients with liver cirrhosis or immunosuppression.

Laryngoscope blades and handles as sources of cross-infection: an integrative review.

The lack of standardization of efficient procedures to clean and disinfect laryngoscope blades and handles, which may be important sources of infection during their clinical use, has been reported previously, revealing contamination with blood, body fluids and micro-organisms. This paper aimed to evaluate the evidence available in the literature regarding the risk of laryngoscope blades and handles as a source of patient contamination. An integrative review of the literature was performed using databases such as Medline, LILACS, SciELO, Cochrane Library, BDENF and PubMed, and keywords in Medical Subject Headings (MeSH). The sample comprised 20 articles published between 1994 and 2012. The studies demonstrated risk of cross-infection and no consensus in current guidelines regarding cleaning and disinfection of this equipment. It was concluded that there are important gaps to be filled and urgent investigations required in order to facilitate standardization of efficient procedures to clean and disinfect laryngoscope blades and handles, and in turn to reduce the potential risk to which the patient and/or health team is exposed.

UriSed as a screening tool for presumptive diagnosis of urinary tract infection.

BACKGROUND: Although a quantitative urine culture is essential for the final diagnosis of urinary tract infection, it is time-consuming and an expensive procedure. Effective screening tests would be a promising alternative to provide immediate results for the clinician and eliminate unnecessary culturing for most of the negative samples. The aim of this study was to evaluate the performance of an automated sediment analyzer (UriSed) as screening tool for presumptive diagnosis of urinary tract infection. METHODS: We studied 1379 fresh midstream clean-catch urine samples from children to elderly. All samples were submitted to automated sediment analysis (UriSed) and quantitative urine culture (CLED medium agar). RESULTS: The sediment analyzer detected leukocyturia and/or significant bacteriuria with sensitivity of 97%, specificity of 59%, positive predictive value of 27%, negative predictive value of 99%, and accuracy of 64% at cutoff values of bacteria count ≥12.6 elements/hpf and WBC ≥6 cells/hpf. These data suggest a potential 52% reduction of unnecessary urine cultures. CONCLUSION: The UriSed seems to be an efficient tool for screening UTI with high sensitivity and low rate of false-negative results.

Changes in vancomycin-resistant Enterococcus faecium causing outbreaks in Brazil.

Enterococci have been implicated in severe human infections as a consequence of associated determinants of virulence and antimicrobial resistance. The majority of vancomycin-resistant Enterococcus faecium (VRE(fm)) connected to outbreaks worldwide pertains to the clonal complex 17 (CC17). In Brazil, the majority of VRE(fm) involved in outbreaks reported so far are not related to CC17. VRE(fm) strains responsible for an outbreak and sporadic cases in hospitals located in the city of Campinas, Brazil, were compared to other VRE(fm) strains in the country. Twenty-two out of 23 E. faecium were vancomycin-resistant and harboured the vanA gene. One vancomycin-susceptible E. faecium (VSE(fm)) strain was included in this study because it was isolated from a patient who one week later harboured a VRE(fm). All strains, except VSE, showed the same alteration in the VanA element characterised by deletion of the left extremity of the transposon and insertion of IS1251 between the vanS and vanH genes. Genes codifying virulence factors such as collagen-adhesin protein, enterococcal surface protein and hyaluronidase were detected in the VRE(fm) and VSE(fm) studied. Both pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) revealed that VRE(fm) and VSE(fm) strains have a clonal relationship. New sequence types (STs) were identified by MLST as ST447, ST448, ST478 and ST412 but all belonged to the CC17. The present study revealed that VRE(fm) outbreaks in Brazil were caused by strains that did not share a common evolutionary history, and that VRE(fm) strains belonging to CC17 could be predominant in Brazil as in other countries.