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Introduction: The difficulty in treating Acinetobacter baumannii infections due to its high rate of resistance to antibiotics has led to the study of mechanisms inherent to the pathogen itself that can be used as effective targets in the treatment. Host Factor I Protein (Hfq) is an RNA chaperone generally necessary to assist in the connection between sRNAs and their mRNA target acting in the regulation of different genes, studies carried out in a range of bacterial species have shown that Hfq acts in a pleiotropic manner, contributing to virulence and response stress. Aim: To summarize current knowledge about the role of the Hfq RNA chaperone in the virulence and antibiotic resistance of Acinetobacter baumannii. Outlining: This is an integrative review developed from articles published in any language on Science Direct and PubMed platforms. Data collection and analysis were carried out between the period of April 2020 and February 2021. Results: Hfq shown to play important roles in cell growth, OMVs, metabolism of carbon sources, tolerance to physical and chemical stress, virulence through biofilm formation, fimbriae modulation, among others. Implications: Our work shows data that strengthen the role of Hfq in different aspects of virulence and environmental adaptation, including antimicrobial resistance of this pathogen, warning about the importance of Hfq as a possible future effective target in the treatment of these infections.

Introduction: Visceral leishmaniasis (VL) is a life-threatening infection remaining as one of the most neglected tropical diseases around the world. Despite scientific advances, an accurate diagnosis of VL remains a challenge. Loop-mediated isothermal amplification (LAMP) has emerged as a promising diagnostic tool with the possibility of becoming a point-of-care test to guide VL diagnosis and treatment.Areas covered: We conducted a systematic review assessing LAMP systems for diagnosing VL from 2000 to 2019. We performed structured searches in PubMed, LILACS, Scopus, and Web of Science without language restriction. Two reviewers screened articles, completed the data extraction and assessment of the risk of bias. A qualitative summary of the included studies was performed.Expert opinion: LAMP could be used as a screening test for VL diagnosis, so tissue aspiration could be performed only for those who are LAMP negative. We recommend more studies about the performance of the Loopamp™ Leishmania Detection kit and the Brazilian LAMP assay. Thus, we expect in the future the constitution of an international consortium to share experiences, projects, and other LAMP approaches mainly among researchers and institutions located within VL endemic countries.

Staphylococcus spp. is one of the major infection-associated bacteria within health care, especially in intensive care units, and one of principal cause of complication in cancer patients. This study compared the antimicrobial susceptibility profile and frequency of resistance (mecA, blaZ, ermA and ermC) and virulence (icaA, icaD and hlg) genes in Staphylococcus spp. from patients of Oncology Hospital (OH) and University Hospital (UH). The type of the ccr complex was assessed by PCR among the mecA positive isolates from the UH. Higher percentage of susceptible isolates, except for oxacillin and cefoxitin was found among the UH isolates and 27,3% vancomycin-resistant isolates were identified through the screening spot; 41 isolates displayed the MLSBc phenotype and five the MSLBi phenotype, and one isolate from the OH displayed the constitutive phenotype ermC gene. The ccr types I and II were identified with a higher frequency of ccr type I. No statistically significant difference was found in the frequency of the genes between the two groups of patients or in the two hospitals. Regarding the virulence genes, there was statistically significant difference when comparing the two hospitals. Staphylococcus spp. es una de las principales bacterias asociadas a las infecciones en la asistencia sanitaria, especialmente en las unidades de cuidados intensivos, y una de las principales causas de complicaciones en los pacientes oncológicos. Este estudio comparó el perfil de susceptibilidad antimicrobiana y la frecuencia de genes de resistencia (mecA, blaZ, ermA y ermC) y virulencia (icaA, icaD y hlg) en Staphylococcus spp. de pacientes de un Hospital Oncológico (OH) y Hospital Universitario (HU). El tipo de complejo ccr se evaluó mediante PCR entre los aislados mecA positivos de la HU. Se encontró un mayor porcentaje de aislados susceptibles, a excepción de oxacilina y cefoxitina, entre los aislados de UH y el 27,3% de los aislados resistentes a la vancomicina se identificaron mediante el método de cribado; 41 aislamientos exhibieron el fenotipo MLSBc y cinco el fenotipo MSLBi, y un aislado OH del fenotipo constitutivo exhibió el gen ermC. Se identificaron CCR tipo I y tipo II, con mayor frecuencia de CCR tipo I. No hubo diferencia estadísticamente significativa en la frecuencia genética entre los dos grupos de pacientes o en los dos hospitales. En cuanto a los genes de virulencia, hubo una diferencia estadísticamente significativa al comparar los dos hospitales. Staphylococcus spp. é uma das principais bactérias associadas a infecções na área de saúde, especialmente em unidades de terapia intensiva, e uma das principais causas de complicações em pacientes com câncer. Este estudo comparou o perfil de susceptibilidade aos antimicrobianos e a frequência de genes de resistência (mecA, blaZ, ermA e ermC) e virulência (icaA, icaD e hlg) em Staphylococcus spp. de pacientes de um Hospital Oncológico (OH) e Hospital Universitário (HU). O tipo de complexo ccr foi avaliado por PCR entre os isolados mecA positivos do HU. Maior porcentagem de isolados suscetíveis, exceto para oxacilina e cefoxitina, foi encontrada entre os isolados de UH e 27,3% dos isolados resistentes à vancomicina foram identificados através método de screening; 41 isolados exibiram o fenótipo MLSBc e cinco o fenótipo MSLBi, e um isolado do OH do fenótipo constitutivo exibiu o gene ermC. Os ccr tipos I e II foram identificados, sendo maior frequência do ccr tipo I. Não foi encontrada diferença estatisticamente significativa na frequência dos genes entre os dois grupos de pacientes ou nos dois hospitais. Em relação aos genes de virulência, houve diferença estatisticamente significativa na comparação dos dois hospitais.

Bloodstream infections remain one of the most common major complications in cancer patients. The aim of the study was to describe the etiology, phenotypic and molecular epidemiology of ICS isolates from cancer patients. Method: identification and the resistance profile were carried out using the automated biochemical method Vitek 2®. The presence and genes resistant to carbapenemases blaIMP, blaVIM, blaGIM, blaSIM, blaSPM, blaKPC, blaNDM, genes oxacilinase blaOXA-48, blaOXA-58, and the presence of ESBL genes blaSHV, blaCTX, blaTEM for Gram-negatives, as well as, mecA, vanA and vanB for Gram-positives were investigated in blood culture isolates. Result: Escherichia coli and Klebsiella pneumoniae were the most frequent pathogens. The serine-β-lactamase gene blaOXA-48 was the most frequent, followed by MβL blaSIM. blaTEM and blaCTX were the most common among ESBL. The mecA and vanA genes were found in Staphylococcus spp. and Enterococcus faecium, respectively. Candida spp showed high resistance to voriconazole and fluconazole. Conclusion: measures to prevent and control the spread of resistance genes are essential to reduce the risks of morbidity and mortality.

The article's abstract is no available.

INTRODUCTION: Pulmonary tuberculosis caused by Mycobacterium tuberculosis is a serious public health problem affecting millions of people worldwide. The development of easy and low-cost diagnostic methods is crucial for disease control in rural remote and poverty areas and among vulnerable groups. OBJECTIVE: To evaluate the accuracy of laboratory methods for the diagnosis of Pulmonary tuberculosis. MATERIAL AND METHODS: Sputum samples from patients with clinical signs and symptoms were analyzed by microscopy after chemical treatment and spontaneous sedimentation and compared with methods employed routinely: direct sputum smear microscopy, culture, and GeneXpert®MTB/RIF. RESULTS: From the samples analyzed, 16% were positive by microscopy in the processed samples, 18% by both culture and Xpert®MTB/RIF, while 13% in the direct microscopy. The processed samples showed a 31% increase in positivity (57 samples) compared to conventional direct microscopy. In the analysis of the accuracy of the evaluated methods, all the results were statistically significant proving that they were not randomly positive or negative and confirming that there is a tendency for these results. CONCLUSION: Chemical treatment and spontaneous sedimentation of the sputum samples procedure represent an effective diagnostic tool in situations where more advanced technologies are not feasible. Besides the higher accuracy and greater detection of positive cases regarding the direct smear, the procedure strengthens biosafety by decreasing the risks of aerial contamination by Mycobacterium tuberculosis for laboratory professionals. RESUMEN INTRODUCCIÓN: La tuberculosis pulmonar causada por Mycobacterium tuberculosis es un grave problema de salud pública que afecta millones de individuos en el mundo. El desarrollo de métodos de diagnóstico fáciles y de bajo costo es esencial para el control de la enfermedad en zonas rurales remotas y pobres y entre los grupos vulnerables. OBJETIVO: Evaluar la exactitud de métodos de laboratorio para el diagnóstico de tuberculosis pulmonar. MATERIAL Y MÉTODO: Las muestras del esputo de pacientes con signos y síntomas clínicos fueron analizadas por microscopía luego de tratamiento químico y sedimentación espontánea y comparados con métodos empleados ordinariamente: baciloscopía directa de esputo, cultivo y GeneXpert® MTB/RIF. RESULTADOS: Entre las muestras analizadas, 16% fueron positivas por microscopía en las muestras procesadas; 18% por cultivo y Xpert® MTB/RIF; y 13% por microscopía directa. Las muestras procesadas presentaran un aumento de 31% de positividad (57 muestras) con respecto a la microscopía directa convencional. En el análisis de los métodos, todos los resultados fueron estadísticamente significativos, comprobando que no eran aleatoriamente positivos o negativos y confirmando que hay una tendencia para esos resultados. CONCLUSIÓN: El tratamiento químico y la sedimentación espontánea de las muestras de esputo representan una herramienta diagnóstica eficaz en las situaciones en las cuales tecnologías más avanzadas no son viables. Además de la mayor precisión y mayor detección de casos positivos de lo que hace el frotis directo, el procedimiento fortalece la bioseguridad, disminuyendo los riesgos de contaminación del aire por Mycobacterium tuberculosis para el personal de laboratorio. RESUMO INTRODUÇÃO: A tuberculose pulmonar causada por Mycobacterium tuberculosis é um grave problema de saúde pública que afeta mundialmente milhões de indivíduos. O desenvolvimento de métodos de diagnóstico fáceis e de baixo custo é essencial para o controle da doença nas áreas rurais remotas e de pobreza e entre os grupos vulneráveis. OBJETIVO: Avaliar a acurácia dos métodos laboratoriais para o diagnóstico de tuberculose pulmonar. MATERIAL E MÉTODOS: As amostras de escarro de pacientes com sinais e sintomas clínicos foram analisadas por microscopia após tratamento químico e sedimentação espontânea e comparadas com métodos empregados rotineiramente: baciloscopia direta do escarro, cultura e GeneXpert® MTB/RIF. RESULTADOS: Das amostras analisadas, 16% foram positivas por microscopia nas amostras processadas; 18%, por cultura e Xpert® MTB/RIF; e 13%, por microscopia direta. As amostras processadas apresentaram um aumento de 31% de positividade (57 amostras) em relação à microscopia direta convencional. Na análise dos métodos avaliados, todos os resultados foram estatisticamente significativos, comprovando que não eram positivos ou negativos aleatoriamente e confirmando que há uma tendência para esses resultados. CONCLUSÃO: O tratamento químico e a sedimentação espontânea das amostras de escarro representam uma ferramenta diagnóstica eficaz nas situações em que tecnologias mais avançadas não são viáveis. Além da maior precisão e maior detecção de casos positivos em relação ao esfregaço direto, o procedimento reforça a biossegurança, diminuindo os riscos de contaminação aérea por Mycobacterium tuberculosis para profissionais de laboratório

Plague, caused by the Yersinia pestis bacterium, has several foci scattered throughout a large area from the Brazilian territory that ranges from the Northeastern State of Ceara to the Southeastern State of Minas Gerais and another separated area at the State of Rio de Janeiro. This review gathers data from plague control and surveillance programs on the occurrence and geographic distribution of rodent hosts and flea vectors in the Brazilian plague areas during the period of from 1952 to 2019. Furthermore, we discuss how the interaction between Y. pestis and some rodent host species may play a role in the disease dynamics. The absence of human cases nowadays in Brazil does not mean that it was eradicated. The dynamics of plague in Brazil and in other countries where it was introduced during the 3rd pandemic are quite alike, alternating epidemics with decades of quiescence. Hence, it remains an important epidemic disease of global concern. The existence of a large animal reservoir and competent vectors demonstrate a need for continuous surveillance to prevent new outbreaks of this disease in humans.

Objective To describe the molecular mechanisms of polymyxins resistance in five Enterobacteriaceae clinical isolates from a tertiary hospital of Recife, Brazil. Methods The species identification and the susceptibility to antimicrobials were firstly performed by automatized methods and polymyxin resistance was confirmed by broth microdilution methods. The genetic basis of resistance was characterized with WGS analyses to study their resistome, plasmidome and mobilome, by BLAST searches on reference databases. Results Five (5%) Enterobacteriaceae isolates, comprising Escherichia coli (n = 2), Klebsiella pneumoniae (n = 2) and Citrobacter freundii (n = 1) species, exhibited polymyxin resistance. The mcr-1.1 gene was found in identical IncX4-plasmids harbored by both K. pneumoniae C119 (PolB MIC = 512 mg/L) and E. coli C153 (PolB MIC = 8 mg/L). The remaining E. coli strain C027 harbored the mcr-5.1 gene on an undefined Inc-plasmid (PolB MIC 256 mg/L). Some amino acid substitutions in PmrA (S29G, G144S), PmrB (S202P; D283G, W350*, Y258N) and PhoP (I44L) was detected among the E. coli clinical isolates, however they were also found in colistin-susceptible strains and predicted as neutral alterations. The mgrB of the ST54 KPC-2-producing K. pneumoniae C151 (PolB MIC = 32 g/mL) was interrupted at 69 nt by the IS903 element. The ST117 C. freundii C156 (PolB MIC = 256 mg/L) showed the A91T substitution on HAMP domain of the histidine kinase sensor CrrB, predicted as deleterious and deemed the remarkable determinant to polymyxins resistance in this strain. Conclusions Diverse mechanisms of polymyxins resistance were identified among clinical Enterobacteriaceae from a tertiary hospital of Recife, Brazil, such as plasmid-mediated MCR-1 and MCR-5; IS903-interruption of mgrB and mutation in CrrAB regulatory system. These findings highlight the involvement of the identified plasmids on mcr dissemination among Enterobacteriaceae; warn about co-selection of the polymyxin-resistant and KPC-producer K. pneumoniae ΔmgrB lineage by carbapenems usage; and demonstrate potential role of CrrAB on emerging of polymyxin resistance among Enterobacteriaceae, besides Klebsiella species.

Acinetobacter baumannii is an opportunistic bacterial pathogen infecting immunocompromised patients and has gained attention worldwide due to its increased antimicrobial resistance. Here, we report a comparative whole-genome sequencing and analysis coupled with an assessment of antibiotic resistance of 46 Acinetobacter strains (45 A. baumannii plus one Acinetobacter nosocomialis) originated from five hospitals from the city of Recife, Brazil, between 2010 and 2014. An average of 3,809 genes were identified per genome, although only 2,006 genes were single copy orthologs or core genes conserved across all sequenced strains, with an average of 42 new genes found per strain. We evaluated genetic distance through a phylogenetic analysis and MLST as well as the presence of antibiotic resistance genes, virulence markers and mobile genetic elements (MGE). The phylogenetic analysis recovered distinct monophyletic A. baumannii groups corresponding to five known (ST1, ST15, ST25, ST79, and ST113) and one novel ST (ST881, related to ST1). A large number of ST specific genes were found, with the ST79 strains having the largest number of genes in common that were missing from the other STs. Multiple genes associated with resistance to β-lactams, aminoglycosides and other antibiotics were found. Some of those were clearly mapped to defined MGEs and an analysis of those revealed known elements as well as a novel Tn7-Tn3 transposon with a clear ST specific distribution. An association of selected resistance/virulence markers with specific STs was indeed observed, as well as the recent spread of the OXA-253 carbapenemase encoding gene. Virulence genes associated with the synthesis of the capsular antigens were noticeably more variable in the ST113 and ST79 strains. Indeed, several resistance and virulence genes were common to the ST79 and ST113 strains only, despite a greater genetic distance between them, suggesting common means of genetic exchange. Our comparative analysis reveals the spread of multiple STs and the genomic plasticity of A. baumannii from different hospitals in a single metropolitan area. It also highlights differences in the spread of resistance markers and other MGEs between the investigated STs, impacting on the monitoring and treatment of Acinetobacter in the ongoing and future outbreaks.

Objetivo: caracterizar aspectos epidemiológicos e clínicos fundamentais, discutir a metodologia do diagnóstico e tecer recomendações sobre as condutas perante a suspeição de casos de peste. Métodos: revisão bibliográfica e levantamento das internações e mortes por peste registradas no Sistema de Informações Hospitalares do Sistema Único de Saúde. Resultados e conclusões: a existência de diagnósticos equivocados de uma doença potencialmente fatal, além dos registros hipotéticos de internações e mortes, constitui um desafio a ser superado, pois espelha uma situação inaceitável, em que um possível e insólito diagnóstico não é investigado e acumula-se nos sistemas de informação.

Introduction: Visceral leishmaniasis (VL) is a life-threatening infection remaining as one of the most neglected tropical diseases around the world. Despite scientific advances, an accurate diagnosis of VL remains a challenge. Loop-mediated isothermal amplification (LAMP) has emerged as a promising diagnostic tool with the possibility of becoming a point-of-care test to guide VL diagnosis and treatment. Areas covered: We conducted a systematic review assessing LAMP systems for diagnosing VL from 2000 to 2019. We performed structured searches in PubMed, LILACS, Scopus, and Web of Science without language restriction. Two reviewers screened articles, completed the data extraction and assessment of the risk of bias. A qualitative summary of the included studies was performed. Expert opinion: LAMP could be used as a screening test for VL diagnosis, so tissue aspiration could be performed only for those who are LAMP negative. We recommend more studies about the performance of the Loopamp™ Leishmania Detection kit and the Brazilian LAMP assay. Thus, we expect in the future the constitution of an international consortium to share experiences, projects, and other LAMP approaches mainly among researchers and institutions located within VL endemic countries.

Aeromonads are mainly opportunistic pathogens; however, many species are emerging as important human pathogens. Therefore, monitoring these bacteria and their accurate characterization of its species is highly important. Aeromonas Aer593 strain was recovered from a diarrhoea outbreak and did not group with any previously described Aeromonas species by housekeeping gene sequencing. To clarify the taxonomic position of Aer593, its genome was sequenced and analysed by multilocus phylogenetic analysis (MLPA), in silico DNA–DNA hybridization (isDDH), average nucleotide identity (ANI) and core genome-based phylogenetic analyzes. The MLPA with the housekeeping genes gyrB, rpoD, recA, dnaJ, gyrA and dnaX ranked the Aer593 isolate into an independent branch suggesting that it could represent a new species. However, the identity percentages of Aer593 to A. caviae strains using robust genomic analysis by isDDH and ANI were at least 81.3% and 97.8%, respectively, defining Aer593 as A. caviae. Multilocus sequence typing (MLST) presented an exact match against only a single allele (groL96) and the novel ST648 was assigned for this strain. The core genome-based phylogenetic analyses with a total of 863 orthologous genes also grouped the Aer593 isolate with A. caviae reference strains. These findings warn about the possibility of misidentification of some Aeromonas strains by MLPA and show that high-resolution genome-wide analysis is essential for the correct identification of ambiguous Aeromonas strains.

Introduction: Plague is an acute, infectious zoonotic disease, primarily of wild rodents and their fleas, that affects humans and other mammals. In Brazil, several plague foci are located in the northeast region. Plague surveillance based on monitoring of rodents was discontinued in 2007, and the current information on rodent populations is unsatisfactory. Our purpose was to update the information on rodents and other small mammals in plague foci in northeastern Brazil. Methodology: Nine surveys in the historically most important northeastern plague areas were conducted in 2013-2015. Results: In this study, 393 animals (13 rodent and four marsupial species) were entrapped. The plague bacterium Yersinia pestis was not detected in tissue sample cultures from the 225 animals that were analyzed. Eighty sera samples were analyzed for anti-F1 antibodies by hemagglutination (HA) and protein A ELISA tests, and all were negative, except for one marsupial, Monodelphis domestica, which was HA positive. Conclusions: Qualitative and quantitative differences in the animal populations were observed in the areas surveyed, and the antibody positive marsupial indicated that plague continues to circulate in the wild.

INTRODUCTION: Microscopic identification of active pulmonary tuberculosis (PTB) from direct smears of sputum (DS) is widely used for detection, but has limited sensitivity. Here, we assessed the yield of acid-fast bacilli (AFB) detection in processed sputum smears (PSS). METHODS: Sputum samples were simultaneously analyzed by direct sputum smearing and after chemical treatment and spontaneous sedimentation. RESULTS: Of the 1,719 samples analyzed, 16.4% were positive for AFB in conventional DS and 21.4% in PSS, corresponding to a 30% increase in detection. CONCLUSIONS: Increased sensitivity from analyzing PSS and better safety protocols will contribute to improved detection and control of the disease.

The presence of methicillin-resistant Staphylococcus aureus (MRSA) strains in food products is a major issue for food safety. The present study was conducted to evaluate the occurrence and antimicrobial resistance profile of S. aureus, focusing on MRSA isolates, in ready-to-eat sashimi from Japanese restaurants in Salvador, Brazil. A total of 127 sashimi samples were collected directly from the take-out service in 16 restaurants. The staphylococcal isolates were identified morphologically and biochemically with standard laboratory procedures. S. aureus isolates were tested with a disk diffusion assay against seven antibiotics, and the cefoxitin and oxacillin were used to identify MRSA strains. Isolates with the MRSA phenotype were confirmed with a PCR assay. S. aureus was found in 73% of the sashimi samples, including sashimi from tuna (75.5% of samples) and salmon (72.5% of samples). Among those positive samples, 37% were contaminated with MRSA strains, found among 38.8% of salmon sashimi and 34.0% of tuna sashimi. Penicillin resistance was the most common type of antimicrobial resistance, found in 65.5% of the sashimi samples, followed by resistance to tetracycline (22.5%), erythromycin (16.0%), and ciprofloxacin (3.2%). Only two S. aureus isolates collected from different fish samples and restaurants had presumed resistance to vancomycin. The high prevalence of S. aureus and MRSA in these sashimi samples indicates a potential risk for foodborne disease, especially MRSA, spreading in the community. HIGHLIGHTS

Visceral leishmaniasis is a serious and debilitating infection with high fatality rate in tropical and subtropical countries. As clinical symptoms of visceral leishmaniasis are not so specific, confirmatory diagnostic methods with high sensitivity and specificity are needed. Noninvasive methods have been developed using urine as a clinical sample for visceral leishmaniasis diagnosis. In fact, there is a clear correlation between kidney impairment and Leishmania DNA in urine. However, it has been proved that Leishmania nucleic acid may also be isolated from patients without any sign of renal involvement. Even though urine has become a promissing biological sample, it is still not widely used due to several issues, such as (i) incomprehension of the whole renal pathophysiology process in visceral leishmaniasis, (ii) presence of many amplification inhibitors in urine, and (iii) lack of an efficient urinary DNA extraction method. In this article, we performed a literature review to bring a new perspective for Leishmania DNA isolation in urine. Keywords: Visceral leishmaniasis, Renal involvement, Diagnosis, Urine sample, DNA extraction

Background The latex from the medicinal plant Calotropis procera is often used in folk medicine against infectious and inflammatory diseases. Purpose In this study, we investigate a protein fraction with immunomodulatory properties, named LPPI, against experimental infections, in vitro and in vivo, with a virulent strain of Listeria monocytogenes. Study design LPPI was exposed to cultured macrophages or Swiss mice and then challenged with L. monocytogenes. Methods Peritoneal macrophages were obtained from Swiss mice, and cultured in 96-well microplates. Soluble latex proteins (LP) were subjected to fractionation by ion-exchange chromatography. The major peak (LPPI) was added into wells at 10 or 100 µg/ml. Albumin (100 µg/ml) was used for comparison between protein treatments. After incubation for 1 h at 5% CO2/ 37°C, the supernatant was discarded and 0.2 ml of L. monocytogenes overnight culture was added in the wells. Following 4 h and 24 h infection, the cytokine mRNA expression was evaluated as well as the number of intracellular colony forming units. Swiss mice (n = 16) were injected intraperitoneally (i.p.) with LPPI (5 and 10 mg/kg) while the control mice received albumin (10 mg/kg) or LP (10 mg/kg). After 24 h, all animal groups were challenged with L. monocytogenes (106 CFU/ ml), also by i.p. route. Results LPPI was not toxic to uninfected macrophages (pMO) and significantly increased mRNA expression of TNF-α, IL-6, IL-1β and iNOS. Following infection, cell viability was reduced by 50% in albumin-treated pMO (control); but only 17% in pMO treated with LPPI at 100 µg/ml. In this case, LPPI increased expression of TNF-α and IL-6 whereas the number of bacterial colony-forming units was reduced 100-fold in comparison to control groups. Swiss mice pretreated with LPPI showed dose-dependent survival rates that reached 80%, while mice that received albumin died 1–3 days after infection. After 24 h infection, leukocyte migration to the infectious foci was high in LPPI-treated mice whereas the number of viable bacteria in the peritoneal fluid, liver and bloodstream were significantly reduced. Conclusion We conclude that LPPI present immunomodulatory properties that are beneficial for prevention of systemic bacterial infections caused by the intracellular bacteria L. monocytogenes.

The Yersinia pestis capsular antigen F1 is widely used in plague laboratory diagnosis. Here, we describe the production of an F1 recombinant protein within reduced time and biosafety requirements. Its evaluation in hemagglutination tests indicated that the recombinant F1 can replace the conventional F1 protein for plague diagnosis.

INTRODUCTION: The increasing reports of vancomycin-resistant Staphylococcus strains (VRS) haves caused concern worldwide, from the laboratory detection to patient management. This study aimed to identify the occurrence of VRS strains among healthcare professionals from a university hospital. METHODS: A total of 102 Staphylococcus sp. isolates from healthcare professionals, obtained in a previous study were evaluated according to standard techniques for VRS detection. RESULTS: After screening inoculation of plates containing 6µg/ml of vancomycin, 19 resistant isolates were identified. The susceptibility profile to other antimicrobials revealed 18 multidrug resistant isolates. The minimum inhibitory concentration (MIC) was determined by E-test and broth microdilution. According to E-tests, of 19 isolates grown in BHI-V6, four isolates presented MIC ≥ 128 µg/ml, seven with MIC ranging from 4 to 8 µg/ml, and eight with MIC ≤ 2µg/ml. By broth microdilution, 14 isolates presented MIC ≤ 2 µg/ml and five with MIC ≥ 16µg/ml. The presence of the gene vanA was determined by PCR in the five resistant isolates, and this gene was detected in one of the strains. Furthermore, among the 19 strains, the gene mecA was found in 13 (39,4%) isolates, including the strain carrying the gene vanA. CONCLUSIONS: Based on these results, we highlight the presence of one strain carrying both vanA and the mecA genes, as well as multidrug-resistant strains colonizing healthcare professionals, and their importance as potential vectors to spread strains carrying resistance genes in the hospital environment.

Acinetobacter baumannii is one of the most frequent Gram-negative opportunistic pathogens associated with hospital-acquired infection worldwide. We briefly describe A. baumannii isolates that were recovered from surrounding ICU bed surfaces, exhibiting multidrug resistance phenotype and belonging to some widely spread clonal complexes of clinical A. baumannii isolates. Keywords: Acinetobacter baumannii, Hospital environment, ICU surface

Introduction: Listeria monocytogenes is an important foodborne pathogen and the 4b serotype is responsible for many cases of human listeriosis reported in Brazil. Several listeriosis outbreaks worldwide have involved a small number of well-defined clonal groups, designated as epidemic clones (ECs). Methodology: We studied 71 strains of serotype 4b, including 25 isolates from human cases of listeriosis and 46 from meat-based foods, collected in Brazil between 1977 and 2010. The presence of ECs (I and II) markers and virulence genes (inlA, inlB, ilnC, inlJ and actA) were evaluated by PCR assay. The genetic relationship of ECs-positive strains was assessed by pulsed field gel electrophoresis. Results: ECI and ECII markers were found both in human and food strains, with 19.7% positive for the ECI marker and 40.8% for ECII. Most strains (97.2%) were positive for the virulence genes that were studied. Nevertheless, the actA gene amplicons showed two distinct sizes, with all ECI positive strains exhibiting a 105bp deletion. Pulsed field gel electrophoresis (PFGE) analysis allowed the recognition of highly related strains, particularly from two outbreaks of neonatal listeriosis in São Paulo State occurred in 1992 and 1997, both ECII-positive; and two ECI strains from a human case (1982) and from bovine meat (2009). Conclusions: The presence of ECs among clinical samples and beef isolates of serotype 4b from some regions of Brazil highlights the need for rigorous control of production procedures. Furthermore, the association of ECII with two nosocomial outbreaks suggests its ability to spread in these settings.

Justificativa e Objetivos: Este estudo teve como objetivo avaliar a presença e resistência de Staphylococcus aureus e estafilococos coagulase-negativos resistentes à oxacilina isolados em superfícies e hemoculturas em uma Unidade de Terapia Intensiva de um hospital de emergência na cidade de Caruaru, Pernambuco, Brasil. Métodos: Trata-se de um estudo descritivo, do tipo transversal, sendo as coletas realizadas a partir das superfícies dos leitos de uma Unidade de Terapia Intensiva (UTI). Paralelamente, amostras de hemoculturas dos respectivos pacientes também foram obtidas. As amostras foram coletadas por amostragem de conveniência e a identificação dos microrganismos isolados se deu através de espectrometria de massas. Resultados: Observou-se uma prevalência maior de bactérias do grupo dos Staphylococcus coagulase negativo-SCN (76,92%) em relação à espécie de Staphylococcus aureus (23,07%). Dentre os microrganismos identificados como resistentes à oxacilina, 12,5% das espécies apresentaram semelhança nas amostras de superfície do leito e hemocultura do mesmo paciente. Conclusões: As bactérias resistentes identificadas reforçam a importância de enfatizar as medidas gerais de higienização do ambiente no que se refere a segurança do paciente nos serviços de saúde. DESCRITORES: Infecção Hospitalar. Contaminação de Equipamentos. Unidades de Terapia Intensiva. Infecções Estafilocócicas.

Background Aeromonas spp. are gram-negative bacteria that can cause a variety of infections in both humans and animals and play a controversial role in diarrhea outbreaks. Our aim was to identify clinical and environmental Aeromonas isolates associated with a cholera outbreak in a northeast county of Brazil at the species level. We also aimed to determine the genetic structure of the bacterial population and the virulence potential of the Aeromonas isolates. Methods and results Analysis based on concatenated sequences of the 16S rRNA and gyrB genes suggested the classification of the 119 isolates studied into the following species: A. caviae (66.9%), A. veronii (15.3%), A. aquariorum (9.3%), A. trota (3.4%), A. hydrophila (3.4%) and A. jandaei (1.7%). One isolate did not fit any Aeromonas species assessed, which might indicate a new species. The haplotype network based on 16S rRNA gene sequences identified 59 groups among the 119 isolates and 26 reference strains, and it clustered almost all A. caviae isolates into the same group. The analysis of the frequency patterns of seven virulence-associated genes (alt, ast, hlyA, aerA, exu, lip, flaA/B) revealed 29 virulence patterns composed of one to seven genes. All the isolates harbored at least one gene, and three of them harbored all seven virulence genes. Conclusion The results emphasize the need to improve local water supply and maintain close monitoring of possible bacterial contamination in the drinking water.

Introduction: Methicillin-resistant Staphylococcus aureus (MRSA) strains have been responsible for many nosocomial outbreaks. Within hospitals, colonized employees often act as reservoirs for the spread of this organism. This study collected clinical samples of 91 patients admitted to the intensive care unit (ICU), hemodialysis/nephrology service and surgical clinic, and biological samples from the nasal cavities of 120 professionals working in those environments, of a University Hospital in Recife, in the State of Pernambuco, Brazil. The main objective of this study was to determine the occurrence and dissemination of methicillin- and vancomycin-resistant Staphylococcus spp. Methods: The isolates obtained were tested for susceptibility to oxacillin and vancomycin and detection of the mecA gene. In addition, the isolates were evaluated for the presence of clones by ribotyping-polymerase chain reaction (PCR). Results: MRSA occurrence, as detected by the presence of the mecA gene, was more prevalent among nursing technicians; 48.1% (13/27) and 40.7% (11/27) of the isolates were from health professionals of the surgical clinic. In patients, the most frequent occurrence of mecA-positive isolates was among the samples from catheter tips (33.3%; 3/9), obtained mostly from the hemodialysis/nephrology service. Eight vancomycin-resistant strains were found among the MRSA isolates through vancomycin screening. Based on the amplification patterns, 17 ribotypes were identified, with some distributed between patients and professionals. Conclusions: Despite the great diversity of clones, which makes it difficult to trace the source of the infection, knowledge of the molecular and phenotypic profiles of Staphylococcus samples can contribute towards guiding therapeutic approaches in the treatment and control of nosocomial infections.

A set of twenty strains of Yersinia pestis isolated during the investigation of an outbreak in a focus of the Araripe, municipality of Exu, state of Pernambuco, Brazil, was analyzed for six VNTR (variable number of tandem repeats) loci. The strains, conserved in the bacterioteca SRP (bacterial collection of the Reference Service in Plague), were reactivated and the identification confirmed by bacteriological bacteriophage susceptibility to anti-plague. Changes in the genome were screened for the presence or absence of virulence genes in plasmids prototypical Y. pestis and High-Pathogenicity Island (HPI) of yersinias by multiplex-PCR. The strains proved to be genetically related by MLVA (analysis of multiple loci of variable number of tandem repeats), which reflects the epidemiological relationship of these isolates.

Rapid identification and characterization of Listeria monocytogenes are required for the food industry, epidemiological studies, and disease prevention and control. However, typing procedures are labor-intensive and time-consuming, and they require technical expertise, a panel of sera and reference culture strains or sophisticated and expensive equipment. To improve upon traditional diagnostic methods for L. monocytogenes we developed and evaluated an efficient procedure for the specific identification of L. monocytogenes and the major pathogenic serotypes of the species based on loop-mediated isothermal amplification (LAMP). Four individual reactions were designed using primers targeting any L. monocytogenes serotypes (LAMP-AS) and the 1/2a (LAMP-1/2a), 1/2b (LAMP-1/2b), and 4b (LAMP-4b) serotypes. The procedure distinguished L. monocytogenes from closely genetically related species and the targeted serotypes. Cross-reactivity with a few rare serotypes isolated from food or clinical samples did not impair the usefulness of the procedure. Thus, our approach constitutes a fast, easy and low-cost alternative for L. monocytogenes diagnosis and serotyping and may be useful for surveillance and epidemiological investigation programs.

Plague caused by Yersinia pestis is one of the infectious diseases subject to the International Health Regulations (IHR). Permanent monitoring of the focal plague areas is mandatory in order to enable prompt control measures to prevent the spread of the disease. Therefore, the availability of efficient diagnosis tests is of paramount importance. Here, we describe a loop-mediated isothermal amplification (LAMP)-based procedure for rapid Y. pestis detection. We constructed a set of LAMP primers, which were used in assays to establish the reaction conditions that would lead to the quick visualization of the results by evaluating the test tube with the naked eye. The primers were specifically designed to target the caf1 gene located on pFra/Tox (pMT), a prototypical plasmid of Y. pestis. The LAMP procedure was performed at 65°C for 45 min in a water bath and allowed for the detection of at least 10 pg of bacterial DNA. Due to its simplicity, specificity, sensitivity and rapidity, the LAMP technique is an additional tool that may be implemented in routine plague diagnoses, especially in emergencies.

Here, we report the isolation of 31 Acinetobacter baumannii strains producing OXA-253 in a single large Brazilian city. These strains belonged to five different sequence types (STs), including 4 STs not previously associated with bla OXA-253 . In all strains, the bla OXA-253 gene was located in a plasmid within a genetic environment similar to what was found previously in Brazil and Italy. The reported data emphasize the successful transmission of the bla OXA-253 gene through a large area and the tendency for this resistance determinant to remain in the A. baumannii population.

Plague outbreaks are occasionally reported in Brazil. Unfortunately, due to great genetic similarity, molecular subtyping of Yersinia pestis strains is difficult. Analysis of multiple-locus variable number of tandem repeats (VNTR), also known as MLVA, has been found to be a valuable tool to discriminate among strains. To check for genetic differences, strains obtained from two different ecological complexes in Brazil collected during two different epidemiological events, an epizootic in Sítio Alagoinha in 1967 and an outbreak in Planalto da Borborema in 1986, were subtyped through MLVA using 12 VNTR loci. Three clusters (A, B and C) were observed. Of the 20 strains from the epizootic, 18 fit into cluster A. Cluster A was divided into two subgroups: A(1) (15 strains) and A(2) (3 strains). Of the 17 strains from the outbreak, 15 fit into cluster B. Cluster B was divided into three subgroups: B(1) (4 strains), B(2) (4 strains) and B(3) (7 strains). Cluster C is a singleton with one epizootic strain. The external standards, Y. pestis CO92 and Y. pseudotuberculosis IP32953, formed two clusters of singletons. The stability of 12 VNTR loci of three unrelated cultures included in this study was assessed. The 12 VNTR loci were stable through multiple serial subcultures in the laboratory. MLVA revealed that Y. pestis populations in Brazil are not monomorphic, and that there is intraspecific genetic diversity among Brazilian plague strains. We conclude that there is some correlation among genetic groups of this species, related to the temporal and geographic origin of isolates.

Through a continuous bacteriological monitoring programme carried out by the Health Secretariat of the State of Pernambuco, Brazil, two isolates of Vibrio cholerae O1 El Tor Ogawa were discovered in an endemic area in 2001, during a cholera inactive period, along with six V. cholerae non-O1/non-O139 strains and two Aeromonas veronii biovar sobria strains showing an unusual characteristic of agglutination with O1 antiserum. Between that time and 2005, eight other O1 isolates were found. The virulence genes present in the V. cholerae differed among strains, with only three O1 strains harboring the ctxA gene. The O1 and some non-O1/non-O139 strains displayed identical patterns of amplification of the 16S-23S intergenic spacer region. RAPD of the 10 V. cholerae O1 strains, with the two primers used, revealed heterogeneity. The presence of V. cholerae carrying virulence genes in the aquatic basins examined confirms that they constitute a vibrio reservoir during a cholera inactive period, thus strengthening the argument for a continuous monitoring programme and preventative measures for cholera, mainly in the areas where the supply of drinking water is deficient.

A multiplex nested PCR method for detection of Vibrio cholerae O1 using a single tube was developed (MSTNPCR). Firstly, single-tube nested PCR (STNPCR) with primers directed to ctx A gene was standardized, and its detection limit was compared to simple PCR and two-step nested PCR. Secondly, primers directed to rfb N gene were added to the reaction. The detection limit of the multiplex reaction was determined using V. cholerae O1 DNA and V. cholerae O1 grown in alkaline peptone water (APW). STNPCR was shown to be approximately 100-fold more sensitive than simple PCR and 10 times less sensitive than two-step nested PCR. This drawback is compensated by a lower risk of cross-contamination. The addition of a second target did not impair the detection limit of STNPCR (as little as 1 pg of V. cholerae O1 DNA detected). MSTNPCR could specifically detect up to three V. cholerae O1 cells or colony forming units (cfu) directly from the APW growth. A diagnostic kit consisting of a set of microtubes having the inner primers fixed onto the inside of the tube cap and a set of tubes containing the reaction mixture was evaluated for stability, and it proved to be stable for five months at − 20 °C. Therefore, MSTNPCR would be useful in the detection of V. cholerae O1 directly from environmental waters in cholera endemic areas and in complementing the identification of toxigenic strains isolated by culture.

UNLABELLED Since its identification as the causative agent of plague in 1894, thousands of Yersinia pestis strains have been isolated and stored. Here, we report the ability of Y. pestis to survive up to 47 years in agar stabs, in rubber-stoppered tubes, under refrigeration (+4 to +10°C), although overall subculture recovery rates were poor and inversely related to the length of time stored. Genetic characterization of virulence gene presence among these subcultures was suggestive of significant variation in the genomic stability of Y. pestis subcultures stored under these conditions. Specifically, we found variation in the presence of plasmid and chromosomal virulence markers (genes pla, lcrV, caf1 and irp2) among multiple subcultures of Y. pestis strains in the 'Collection of Yersinia pestis' (Fiocruz-CYP) maintained by the SRP of FIOCRUZ-PE in Brazil. This variation, together with all of the inherent temporal, geographic and other genetic variation represented by all of the recoverable strains in this historical collection was preserved in new frozen culture stocks stored at -70°C as a result of this study. These frozen culture stocks represent a valuable resource for future comparative studies of Y. pestis. SIGNIFICANCE AND IMPACT OF THE STUDY We report the ability of Yersinia pestis to survive up to 47 years in agar stabs, in rubber-stoppered tubes, under refrigeration (+4 to +10°C), although overall subculture recovery rates were poor and inversely related to the length of time stored. Genetic characterization of virulence gene presence among these subcultures was suggestive of significant variation in the genomic stability of Y. pestis subcultures stored under these conditions. This variation, together with all of the inherent temporal, geographic and other genetic variation represented by all of the recoverable strains in the historical 'Collection of Yersinia pestis' (Fiocruz-CYP) maintained by the SRP of FIOCRUZ-PE in Brazil was preserved in new frozen culture stocks stored at -70°C as a result of this study. These frozen culture stocks represent a valuable resource for future comparative studies of Y. pestis.

One hundred seventy nine Vibrio cholerae non-O1/non-O139 strains from clinical and different environmental sources isolated in Brazil from 1991 to 2000 were serogrouped and screened for the presence of four different virulence factors. The Random Amplification of Polymorphic DNA (RAPD) technique was used to evaluate the genetic relatedness among strains. Fifty-four different serogroups were identified and V. cholerae O26 was the most common (7.8%). PCR analysis for three genes (ctxA, zot, ace) located of the CTX genetic element and one gene (tcpA) located on the VPI pathogenicity island showed that 27 strains harbored one or more of these genes. Eight (4.5%) strains possessed the complete set of CTX element genes and all but one of these belonged to the O26 serogroup suggesting that V. cholerae O26 has the potential to be an epidemic strain. The RAPD profiles revealed a wide variability among strains and no genetic correlation was observed.Cento e setenta e nove amostras de V. cholerae não O1/não O139, isoladas de casos clínicos (139) e de meio ambiente (40), no período de 1991 a 2000 no Brasil, foram caracterizadas antigenicamente pelo National Institute of Health (Japão) e investigadas quanto ao seu potencial genético de virulência, representado pelos genes ctxA, zot, ace e tcpA. As análises fenotípicas revelaram extraordinária diversidade antigênica, com a ocorrência de 54 diferentes sorogrupos, com prevalência para O26 (7,8%). A técnica de PCR, empregada na detecção dos genes localizados no elemento genético CTX (ctxA, zot, ace) e na Ilha de Patogenicidade de Vibrio-VPI (tcpA), possibilitou a identificação de 27 cepas contendo qualquer um desses genes. O gene ctxA (codificador da sub-unidade A de CT), só foi evidenciado no sorogrupo O26, sendo também o único capaz de se apresentar com o cassete de virulência de forma intacta. Com base nos resultados obtidos deste estudo preliminar, admite-se a hipótese da potencialidade destas cepas, evoluir para raças epidêmicas.

The pigmentation (pgm) locus is a large unstable area of the Yersinia pestis chromosome composed of a segment of iron acquisition (HPI) linked to a pigmentation segment. In this work we examined the mobility of HPI and the pigmentation segment in three Y. pestis isolates using successive subcultures on Congo red agar (CRA) plates. Strain P. CE 882 was shown to be highly stable while strains P. Exu 340 and P. Peru 375 dissociated into several phenotypes, PCR analysis showing evidence of changes in the pgm locus of the derived cultures. Strains P. Exu 340 and P. Peru 375 produced previously unreported cultures positive for the pesticin/yersiniabactin outer membrane receptor (psn+) but negative for the iron-regulated protein (irp2-), suggesting the occurrence of rearrangements in this chromosomal region and either a sequential loss or the loss of separated segments. These results provide evidence that besides deletion en bloc, specific rearrangements are also involved in the deletion events for that locus.

Introduction The number of reports of intestinal infections caused by Aeromonas spp. has increased significantly in recent years. In most clinical laboratories, identification of these bacteria is carried out by general phenotypic tests that sometimes do not accurately differentiate Aeromonas and Vibrio. Methods A duplex-polymerase chain reaction (PCR) was developed directed to 2 targets identifying Aeromonas spp. pathogenic to humans. Results The duplex-PCR results were reproducible and specific for Aeromonas spp. pathogenic to humans. Conclusions This method will allow differentiation between Vibrio and Aeromonas spp. in patients with in cholera-like symptoms and can also be used in water quality monitoring.

O estudo foi desenvolvido com o objetivo de analisar o perfil plasmidial, pesquisar genes de virulência e identificar os perfis genéticos de 31 cepas de Vibrio cholerae não O1 isoladas de zooplâncton dos estuários dos rios Anil e Bacanga em São Luis MA. O estudo do DNA plasmidial revelou a presença de 2 a 3 plasmídeos em 10 cepas, com pesos moleculares variando de 5,5 a 40 kilobases. A ribotipagem revelou um perfil comum a todas as cepas. A amplificação do DNA genômico por PCR não revelou os genes ctxA, ace e zot, mostrando tratar-se de cepas não patogênicas, enquanto a RAPD-PCR identificou múltiplos perfis genéticos, achado compatível com o grande potencial de variabilidade desta espécie.The study was developed to analyze the plasmidial DNA, research virulence genes and identify genetic diversity of 31 strains of Vibrio cholerae non-O1 isolated from zooplankton of the Bacanga and Anil rivers in São Luis-MA. The study of plasmidial DNA showed 2 or 3 plasmids from 10 strains between 5.5 and 40 kilobasis. There was only single ribotype pattern. PCR methods did not show the genes ctxA, ace and zot, while RADP-PCR identified genetic diversity in the strains, showing the potential for variability in this species.

Optimization of the RAPD reaction for characterizing Salmonella enterica serovar Typhi strains was studied in order to ensure the reproducibility and the discriminatory power of this technique. Eight Salmonella serovar Typhi strains isolated from various regions in Brazil were examined for the fragment patterns produced using different concentrations of DNA template, primer, MgCl2 and Taq DNA polymerase. Using two different low stringency thermal cycle profiles, the RAPD fingerprints obtained were compared. A set of sixteen primers was evaluated for their ability to produce a high number of distinct fragments. We found that variations associated to all of the tested parameters modified the fingerprinting patterns. For the strains of Salmonella enterica serovar Typhi used in this experiment, we have defined a set of conditions for RAPD-PCR reaction, which result in a simple, fast and reproducible typing method.A otimização da reação de RAPD para a caracterização de cepas de Salmonella enterica sorovar Typhi foi estudada com o objetivo de assegurar a reprodutibilidade e o poder discriminatório desta técnica. Oito cepas de Salmonella sorovar Typhi isoladas de algumas regiões do Brasil foram usadas para examinar os padrões de fragmentação produzidos quando foram empregadas concentrações diferentes do DNA molde, do iniciador, do MgCl2 e da enzima Taq DNA polimerase. Com a utilização de dois diferentes perfis de ciclos termais de baixa estringência, foram comparados os padrões de bandeamento obtidos. Um conjunto de dezesseis iniciadores foi avaliado quanto à capacidade de produzir elevado número de fragmentos distintos. Observou-se que variações associadas a todos os parâmetros testados modificaram os padrões de bandeamento. Para as amostras de Salmonella enterica sorovar Typhi utilizadas neste experimento, definiu-se um conjunto de condições para a reação de RAPD-PCR que resultou num método de tipagem simples, rápido e reprodutível.

Optimization of the RAPD reaction for characterizing Salmonella enterica serovar Typhi strains was studied in order to ensure the reproducibility and the discriminatory power of this technique. Eight Salmonella serovar Typhi strains isolated from various regions in Brazil were examined for the fragment patterns produced using different concentrations of DNA template, primer, MgCl2 and Taq DNA polymerase. Using two different low stringency thermal cycle profiles, the RAPD fingerprints obtained were compared. A set of sixteen primers was evaluated for their ability to produce a high number of distinct fragments. We found that variations associated to all of the tested parameters modified the fingerprinting patterns. For the strains of Salmonella enterica serovar Typhi used in this experiment, we have defined a set of conditions for RAPD-PCR reaction, which result in a simple, fast and reproducible typing method.

Aims: To investigate whether modifications in Yersinia pestis isolates from three plague foci from the state of Ceara, Brazil, had occurred over the years as a consequence of genetic adaptation to the environment. Methods and Results: The isolates were studied with respect to susceptibility to antimicrobial drugs, plasmid and protein profiling, pigmentation on Congo red-agar plates, and the presence of some pathogenicity genes using PCR. Most of the expected virulence markers were detected in the cultures examined. There was no evidence of any alteration that could be associated with their origin (patients, rodents and fleas) or period of isolation (1971–1997). Conclusions, Significance and Impact of the Study: Phenotypic or genotypic changes were not detected in the cultures examined. However, the results obtained will serve as a reference to follow the evolution of Y. pestis in these foci.

As atividades de vigilância sorológica da peste nos focos do Estado do Ceará detectaram elevação do número de animais indicadores/sentinela com anticorpos antipestosos a partir de 1995, com pico em 1997 evidenciando aumento da circulação do bacilo pestoso em todos os focos investigados. De um total de 110.725 amostras de soro obtidas de roedores (7.873) e carnívoros domésticos (102.852) analisadas pela técnica de hemaglutinação (HA) para detecção de anticorpos contra o antígeno F1 da Yersinia pestis, 905 revelaram-se positivas, sendo 15 de roedores (4 Rattus rattus e 11 Galea spp), 720 de cães e 170 de gatos. Dos 652 casos humanos suspeitos e contatos investigados apenas dois foram positivos pela HA e um terceiro paciente foi positivo por hemocultura. A cepa isolada revelou-se altamente patogênica para animais de laboratório e mostrou sensibilidade aos antimicrobianos usados no tratamento dos doentes.

As atividades de vigilância sorológica da peste nos focos do Estado do Ceará detectaram elevação do número de animais indicadores/sentinela com anticorpos antipestosos a partir de 1995, com pico em 1997 evidenciando aumento da circulação do bacilo pestoso em todos os focos investigados. De um total de 110.725 amostras de soro obtidas de roedores (7.873) e carnívoros domésticos (102.852) analisadas pela técnica de hemaglutinação (HA) para detecção de anticorpos contra o antígeno F1 da Yersinia pestis, 905 revelaram-se positivas, sendo 15 de roedores (4 Rattus rattus e 11 Galea spp), 720 de cães e 170 de gatos. Dos 652 casos humanos suspeitos e contatos investigados apenas dois foram positivos pela HA e um terceiro paciente foi positivo por hemocultura. A cepa isolada revelou-se altamente patogênica para animais de laboratório e mostrou sensibilidade aos antimicrobianos usados no tratamento dos doentes.Serological surveillance activities regarding the foci of plague in Ceará State have detected a rising number of sentinel animals with antiplague antibodies in 1995, with a peak in 1997 demonstrating an increase in the plague bacteria activities throughout all the foci investigated. From a total of 110,725 serum samples collected from rodents (7,873) and domestic carnivores (102,852) analyzed by the Hemaglutination technique (HA) for antibodies against F1 antigen of Yersinia pestis 905 samples tested positive. In these samples there were 15 rodents (4 Rattus rattus and 11 Galea spp), 720 dogs and 170 cats. Of the 652 human suspected and contact cases investigated by HA, only two were positive. A third case had a positive hemoculture for Y. pestis. The isolate is highly pathogenic for laboratory animals and showed sensitivity to the antimicrobial drugs used for plague treatment.

Eighty-five isolates of Klebsiella pneumoniae and Enterobacter spp., originating from hospital- and community-acquired infections and from oropharyngeal and faecal microbiota from patients in Recife-PE, Brazil, were analyzed regarding the presence of irp2 gene. This is a Yersinia typical gene involved in the synthesis of siderophore yersiniabactin. DNA sequencing confirmed the identity of irp2 gene in five K. pneumoniae, five Enterobacter aerogenes and one Enterobacter amnigenus isolates. To our knowledge in the current literature, this is the first report of the irp2 gene in E. amnigenus, a species considered an unusual human pathogen, and in K. pneumoniae and E. aerogenes isolates from the normal microbiota and from community infections, respectively. Additionally, the analyses of nucleotide and amino acid sequences suggest the irp2 genes derived from isolates used in this study are more closely related to that of Yersinia pestis P.CE882 than to that of Yersinia enterocolitica 8081. These data demonstrated that K. pneumoniae and Enterobacter spp. from normal microbiota and from community- and hospital-acquired infections possess virulence factors important for the establishment of extra-intestinal infections.

Visando complementar as informações sobre a qualidade microbiológica do leite comercializado na cidade do Recife, foram analisadas 250 amostras de leite pasteurizado tipo C e 50 amostras de leite cru para a pesquisa de Yersinia enterocolitica e Listeria monocytogenes, bactérias patogênicas capazes de se desenvolverem em temperatura de refrigeração. Y. enterocolitica não foi encontrada em nenhuma das amostras analisadas, entretanto foi detectada a presença de Y. intermedia e Y. frederiksenii, espécies ambientais que se comportam como patógenos oportunistas. L. monocytogenes também não foi encontrada, mas, através da metodologia empregada para seu isolamento foi obtido um isolamento de Salmonella Montevideo em uma amostra de leite pasteurizado e outro em leite cru. Além dessas, várias outras bactérias foram encontradas, supondo-se que a ampla microbiota crescida nos meios empregados pode ter interferido no isolamento da Y. enterocolitica e L. monocytogenes. In order to improve information about the microbiological quality of the milk commercially available in the city of Recife, 250 samples of pasteurized type-C milk and 50 samples of raw milk were analyzed for Yersinia enterocolitica and Listeria monocytogenes and verify the possible occurrence of Yersinia enterocolitica and Listeria monocytogenes. These bacteria can develop in refrigeration temperatures and are responsible for food-born diseases. Neither Y. enterocolitica nor L. monocytogenes were found in the samples analyzed. However, the presence of Y. intermedia and Y. frederiksenii was detected, these environmental species behave as opportunist pathogens. Through the methodology used for Listeria isolation, one isolate of Salmonella Montevideo was obtained from a sample of pasteurized milk and another isolated from one sample of raw milk. Besides these, several other bacteria species were found. It is likely that the large microbiota present in the samples and the procedures employed to destroy it could have hindered the isolation of Y. enterocolitica and L. monocytogenes.

Visando complementar as informações sobre a qualidade microbiológica do leite comercializado na cidade do Recife, foram analisadas 250 amostras de leite pasteurizado tipo C e 50 amostras de leite cru para a pesquisa de Yersinia enterocolitica e Listeria monocytogenes, bactérias patogênicas capazes de se desenvolverem em temperatura de refrigeração. Y. enterocolitica não foi encontrada em nenhuma das amostras analisadas, entretanto foi detectada a presença de Y. intermedia e Y. frederiksenii, espécies ambientais que se comportam como patógenos oportunistas. L. monocytogenes também não foi encontrada, mas, através da metodologia empregada para seu isolamento foi obtido um isolamento de Salmonella Montevideo em uma amostra de leite pasteurizado e outro em leite cru. Além dessas, várias outras bactérias foram encontradas, supondo-se que a ampla microbiota crescida nos meios empregados pode ter interferido no isolamento da Y. enterocolitica e L. monocytogenes.In order to improve information about the microbiological quality of the milk commercially available in the city of Recife, 250 samples of pasteurized type-C milk and 50 samples of raw milk were analyzed for Yersinia enterocolitica and Listeria monocytogenes and verify the possible occurrence of Yersinia enterocolitica and Listeria monocytogenes. These bacteria can develop in refrigeration temperatures and are responsible for food-born diseases. Neither Y. enterocolitica nor L. monocytogenes were found in the samples analyzed. However, the presence of Y. intermedia and Y. frederiksenii was detected, these environmental species behave as opportunist pathogens. Through the methodology used for Listeria isolation, one isolate of Salmonella Montevideo was obtained from a sample of pasteurized milk and another isolated from one sample of raw milk. Besides these, several other bacteria species were found. It is likely that the large microbiota present in the samples and the procedures employed to destroy it could have hindered the isolation of Y. enterocolitica and L. monocytogenes.

Yersinia pestis, the etiologic agent of plague, harbors three well-characterized plasmids: pFra (90-110kb), pYV (70 kb) and pPst (9.5 kb). Furthermore, some extra-cryptic DNA bands have been observed in a number of wild strains from several foci of the world. Additional bands have also been reported in Brazilian strains. Looking for any relationship among these cryptic DNA bands and the three-prototypical plasmids, we analyzed twelve strains displaying different plasmid content. The DNA bands were hybridized by southern blot with probes directed at the genes caf1, lcrV and pla located respectively on the plasmids pFra, pYV and pPst. The probes were constructed by PCR amplification and labeled with digoxigenin. The Pla probe hybridized with its target (pPst) and with bands of about 35 kb suggesting some homology among them. The Caf1 probe hybridized with the target (pFra) as well as with higher bands. The LcrV also hybridized with the target (pYV) and both with the bands higher than pFra and the bands between pFra and pYV. These results suggest that the large-cryptic bands could represent some rearrangement, open circular or linearized forms of the pFra and pYV plasmids.Yersinia pestis, o agente causador da peste, possui três plasmídios bem caracterizados: pFra (90-110 kb), pYV (70 kb) e pPst (9.5 kb). Adicionalmente, algumas bandas extras de DNA críptico têm sido observadas em numerosas cepas selvagens em vários focos do mundo. Bandas extras também foram observadas em cepas brasileiras. Para verificar se existe alguma homologia entre as bandas extras de DNA críptico e os três plasmídios típicos, foram analisadas 12 culturas de Y. pestis através de hibridização com sondas dirigidas aos genes caf1, lcrV e pla localizados respectivamente nos plasmídios pFra, pYV e pPst. As sondas foram construídas através de amplificação por PCR e marcadas com digoxigenina. A sonda Pla reconheceu seu alvo (pPst) e bandas de cerca de 35 kb sugerindo que estas últimas podem se tratar de um multímero do pPst. A sonda Caf1 reconheceu seu alvo (pFra) assim como bandas mais altas. A sonda LcrV, além de reconhecer seu alvo (pYV), também hibridizou com bandas maiores que pFra e bandas de tamanho entre as de pFra e pYV. Estes resultados sugerem que as bandas grandes poderiam ser resultantes de algum rearranjo, formas abertas circulares ou linearizadas dos plasmídios pFra e pYV.