DETECTION OF Leptospira spp. AND Brucella abortus ANTIBODIES IN FREE-LIVING JAGUARS (Panthera onca) IN TWO PROTECTED AREAS OF NORTHERN PANTANAL, BRAZIL

 This study aimed to assess the exposure of free-living jaguars (Panthera onca) to Leptospira spp. and Brucella abortus in two conservation units in the Pantanal of Mato Grosso, Brazil. The presence of antibodies in blood samples of eleven jaguars was investigated using autochthonous antigens isolated in Brazil added to reference antigen collection applied to diagnosis of leptospirosis by Microscopic Agglutination Test (MAT). The Rose Bengal test was applied for B. abortus antibodies. Two (18.2%) jaguars were seroreactive for the Leptospira spp. antigen and the serovar considered as most infective in both animals was a Brazilian isolate of serovar Canicola (L01). All jaguars were seronegative for B. abortus. These data indicate that the inclusion of autochthonous antigens in serological studies can significantly increase the number of reactive animals, as well as modify the epidemiological profile of Leptospira spp. infection.

OCCURRENCE OF Blastocystis spp. IN UBERABA, MINAS GERAIS, BRAZIL

Intestinal parasites are a problem for public health all over the world. The infection with Blastocystis, a protozoan of controversial pathogenicity, is one of the most common among them all. In this study, the occurrence of intestinal parasites, with emphasis on Blastocystis, in patients at the Universidade Federal do Triângulo Mineiro was investigated in Uberaba (MG) through microscopy of direct smears and fecal concentrates using Ritchie’s method. Feces of 1,323 patients were examined from April 2011 to May 2012. In 28.7% of them at least one intestinal parasite was identified, and the most frequent organisms were Blastocystis spp. (17.8%) and Giardia intestinalis (7.4%). The occurrence of parasitism was higher in children aged 6 -10 years old, and the infection with Blastocystis spp. was higher above the age of six (p < 0.001). The exclusive presence of G. intestinalis and of Blastocystis spp. was observed in 5.4% and 12.2% of the patients, respectively. Regarding patients with diarrheic feces, 8% revealed unique parasitism of Blastocystis spp. Other intestinal parasites observed in children were Ascaris lumbricoides(0.3%) and Entamoeba histolytica/dispar/moshkovskii (1.4%). The Ritchie’s method was more sensitive (92.8%) when compared to direct microscopy (89.8%), with high agreement between them (97.7%, kappa = 0.92). In conclusion, the occurrence of Blastocystis spp. in Uberaba is high and the presence of diarrheic feces with exclusive presence of the parasite of Blastocystis spp. was observed.

MOLECULAR IDENTIFICATION AND ANTIMICROBIAL RESISTANCE PATTERN OF SEVEN CLINICAL ISOLATES OF Nocardia spp. IN BRAZIL

Nocardia is a ubiquitous microorganism related to pyogranulomatous infection, which is difficult to treat in humans and animals. The occurrence of the disease is on the rise in many countries due to an increase in immunosuppressive diseases and treatments. This report of cases from Brazil presents the genotypic characterization and the antimicrobial susceptibility pattern using the disk-diffusion method and inhibitory minimal concentration with E-test® strips. In summary, this report focuses on infections in young adult men, of which three cases were cutaneous, two pulmonary, one neurological and one systemic. The pulmonary, neurological and systemic cases were attributed to immunosuppressive diseases or treatments. Sequencing analysis of the 16S rRNA segments (1491 bp) identified four isolates of Nocardia farcinica, two isolates of Nocardia nova and one isolate of Nocardia asiatica. N. farcinica was involved in two cutaneous, one systemic and other pulmonary cases; N. nova was involved in one neurological and one pulmonary case; and Nocardia asiatica in one cutaneous case. The disk-diffusion antimicrobial susceptibility test showed that the most effective antimicrobials were amikacin (100%), amoxicillin/clavulanate (100%), cephalexin (100%) and ceftiofur (100%), while isolates had presented most resistance to gentamicin (43%), sulfamethoxazole/trimethoprim (43%) and ampicillin (29%). However, on the inhibitory minimal concentration test (MIC test), only one of the four isolates of Nocardia farcinica was resistant to sulfamethoxazole/trimethoprim.

WATER FROM DIFFERENT SOURCES USED FOR THE IRRIGATION OF VEGETABLES TO BE MARKETED: RESEARCH ON Cryptosporidium spp., Giardia spp., AND COLIFORMS IN PARANA, BRAZIL

SUMMARY The aim of this work was to compare, from a parasitological ( Cryptosporidiumspp. and Giardia duodenalis), bacteriological (total and thermotolerants coliforms) and physicochemical perspective, water sources used for drinking and irrigation of vegetables intended to be sold for human consumption. From January 2010 to May 2011, samples of different water sources from vegetable producing properties were collected; 100 liters for parasitological analysis, 200 mL for bacteriological analysis, and five liters for physicochemical analysis. Water samples were filtered under vacuum with a kit containing a cellulose acetate membrane filter, 1.2 µm (Millipore(r), Barueri, SP, Brazil). The material retained on the membrane was mechanically extracted and analyzed by direct immunofluorescence (Merifluor(r)kit). From 20 rural properties investigated, 10 had artesian wells (40 samples), 10 had common wells (40 samples), and one had a mine (four samples), the latter contaminated by Cryptosporidiumspp. In samples from artesian wells, 90 to 130 meters depth, 42.5% were positive for total coliforms and 5.0% were identified to have abnormal coloration. From the samples of common wells, 14 to 37 meters depth, 87.5% were contaminated with total coliforms, 82.5% were positive for thermotolerant coliforms, and 12.5% had color abnormalities. We did not detect the presence of Giardiaspp. or Cryptosporidiumspp. in artesian and common wells. The use of artesian or common wells is an important step in the control of the spreading of zoonoses, particularly Cryptosporidiumspp. and Giardiaspp., as well as artesian wells for coliform control in local production of vegetables to be marketed.

Pseudomonas spp. ISOLATED FROM THE ORAL CAVITY OF HEALTHCARE WORKERS FROM AN ONCOLOGY HOSPITAL IN MIDWESTERN BRAZIL

This cross-sectional study, performed in an oncology hospital in Goiania, aimed to characterize the prevalence of oral colonization and antimicrobial susceptibility of Pseudomonas spp. isolated from the saliva of healthcare workers. Microorganisms were subjected to biochemical tests, susceptibility profile, and phenotypic detection. Of 76 participants colonized with Gram negative bacilli, 12 (15.8%) harbored Pseudomonas spp. Of all isolates, P. aeruginosa (75.0%), P. stutzeri (16.7%), and P. fluorescens (8.3%), were resistant to cefoxitin, and therefore likely to be AmpC producers. The results are clinically relevant and emphasize the importance of surveillance to minimize bacterial dissemination and multiresistance.

SEROLOGICAL DETECTION OF HEPATITIS A VIRUS IN FREE-RANGING NEOTROPICAL PRIMATES (Sapajus spp., Alouatta caraya) FROM THE PARANÁ RIVER BASIN, BRAZIL

Nonhuman primates are considered as the natural hosts of Hepatitis A virus (HAV), as well as other pathogens, and can serve as natural sentinels to investigate epizootics and endemic diseases that are of public health importance. During this study, blood samples were collected from 112 Neotropical primates (NTPs) (Sapajus nigritus and S. cay, n = 75; Alouatta caraya, n = 37) trap-captured at the Paraná River basin, Brazil, located between the States of Paraná and Mato Grosso do Sul. Anti-HAV IgG antibodies were detected in 4.5% (5/112) of NTPs, specifically in 6.7% (5/75) of Sapajus spp. and 0% (0/37) of A. caraya. In addition, all samples were negative for the presence of IgM anti-HAV antibodies. These results suggest that free-ranging NTPs were exposed to HAV within the geographical regions evaluated.

SIXTEEN YEARS OF DERMATOMYCOSIS CAUSED BY Candida spp. IN THE METROPOLITAN AREA OF PORTO ALEGRE, SOUTHERN BRAZIL

The yeasts of the genus Candida infect skin, nails, and mucous membranes of the gastrointestinal and the genitourinary tract. The aim of this study was to determine the prevalence of dermatomycoses caused by Candida spp., and their etiological aspects in the metropolitan area of Porto Alegre, Brazil. A retrospective study with data obtained from tertiary hospital patients, from 1996 to 2011, was performed. The analyzed parameters were date, age, gender, ethnicity, anatomical region of lesions, and the direct examination results. For all the statistical analyses, a = 0.05 was considered. Among positive results in the direct mycological examination, 12.5% of the total of 4,815 cases were positive for Candida spp. The angular coefficient (B) was -0.7%/ year, showing a decrease over the years. The genus Candida was more prevalent in women (15.9% of women versus 5.84% of men), and in addition, women were older than men (54 versus 47 years old, respectively). There was no difference between ethnic groups. The nails were more affected than the skin, with 80.37% of the infections in the nails (72.9% in fingernails and 7.47% in toenails). Our study corroborates the literature regarding the preference for gender, age, and place of injury. Moreover, we found a decrease in infection over the studied period.

HIGH PREVALENCE OF Blastocystis spp. INFECTION IN CHILDREN AND STAFF MEMBERS ATTENDING PUBLIC URBAN SCHOOLS IN SÃO PAULO STATE, BRAZIL

After a gastroenteritis outbreak of unknown etiology in the municipality ofSebastião da Grama, SãoPaulo, Brazil, we conducted a parasitological survey to establish the epidemiological profile of enteroparasitosis in children and staff members attending the public urban schools in operation in town. The cross-sectional study evaluated 172 children aged 11 months to 6 years old and 33 staff members aged 19 to 58 years old. Overall, 96 (55.81%) children and 20 (60.61%) staff members were mono-parasitized, while 58 (33.72%) children and 4 (12.12%) workers were poly-parasitized. Protozoa (88.37%; 72.73%) was more prevalent than helminthes (3.48%; 0%) in children and staff members respectively.Blastocystis spp. was the most prevalent parasite in children (86.63%) and staff members (66.67%). The age of 1 year old or less was found to be associated with increased prevalence of giardiasis [OR = 13.04; 95%CI 2.89-58.91; p = 0.00] and public garbage collection was identified as a protective factor against intestinal helminth infections [OR = 0.06; 95%CI 0.00-0.79; p = 0.03]. Although most of the children tested positive for Blastocystis spp. and also presented clinical signs/symptoms (62.2%), this association was not statistically significant [OR = 1.35; 95%CI 0.53-3.44; p = 0.51]. Intestinal parasites still represent a public health concern and this study underscores the importance of further investigations to better understand the pathogenic role ofBlastocystis spp.

PERFORMANCE OF CONVENTIONAL PCRs BASED ON PRIMERS DIRECTED TO NUCLEAR AND MITOCHONDRIAL GENES FOR THE DETECTION AND IDENTIFICATION OF Leishmania spp.

In visceral leishmaniasis, the detection of the agent is of paramount importance to identify reservoirs of infection. Here, we evaluated the diagnostic attributes of PCRs based on primers directed to cytochrome-B (cytB), cytochrome-oxidase-subunit II (coxII), cytochrome-C (cytC), and the minicircle-kDNA. Although PCRs directed to cytB, coxII, cytC were able to detect different species of Leishmania, and the nucleotide sequence of their amplicons allowed the unequivocal differentiation of species, the analytical and diagnostic sensitivity of these PCRs were much lower than the analytical and diagnostic sensitivity of the kDNA-PCR. Among the 73 seropositive animals, the asymptomatic dogs had spleen and bone marrow samples collected and tested; only two animals were positive by PCRs based on cytB, coxII, and cytC, whereas 18 were positive by the kDNA-PCR. Considering the kDNA-PCR results, six dogs had positive spleen and bone marrow samples, eight dogs had positive bone marrow results but negative results in spleen samples and, in four dogs, the reverse situation occurred. We concluded that PCRs based on cytB, coxII, and cytC can be useful tools to identify Leishmania species when used in combination with automated sequencing. The discordance between the results of the kDNA-PCR in bone marrow and spleen samples may indicate that conventional PCR lacks sensitivity for the detection of infected dogs. Thus, primers based on the kDNA should be preferred for the screening of infected dogs.

Níveis de sensibilidade de enterobactérias, em particular Salmonella typhi, a rifamicina S.V.

Em face à predominante eliminação biliar da rifamicina S.V. atingindo concentrações muitas vêzes superiores aos níveis séricos obtidos com as doses terapêuticas, e pelo possível interêsse dessa verificação para o tratamento dos portadores biliares crônicos de Salmonella typhi determinou-se a concentração mínima inibitória de 165 estirpes de enterobactérias, incluindo 77 amostras de S. typhi. Foi verificado que a maioria das cepas de Escherichia coli, Shigella e Proteus mirabilis correspondiam a uma concentração inibitória mínima entre 33 a 65 μg/ml. Entre 65 e 128 μg/ml foram determinadas as concetrações inibitórias mínimas da maioria das outras espécies de Proteus, de Providencia e de Klebsiella. Para Salmonella e Enterobacter o limite mínino de sensibilidade foi, em regra, igual ou superior a 128 μg/ml. Diferenças mais acentuadas de comportamento entre as enterobactérias foram observadas quanto à ação bactericida da rifamicidas S.V. De uma maneira geral, para E. coli e Shigella, as concentrações inibitórias mínimas já referidas. Para as espécies de Proteus e Providencia houve variação maior de comportamento, mas tendência a que o efeito bactericidas fôsse encontrado em concentrações que correspondiam a 4 vêzes as bacteriostáticas para as mesmas espécies. Finalmente, de modo pouco feliz para os propósitos visados, em Salmonella, com a inclusão de S. typhi, não foi atingido um efeito bactericida, com as mais altas concentrações usadas as quais corresponderam em média a 6 vêzes as concentrações bacteriostáticas para esse gênero.

Tipos bioquímicos de Salmonella typhi de algumas regiões do Brasil

Foram estudadas 813 amostras de Salmonella typhi provenientes dos estados da Bahia, Guanabara, Pará, Pernambuco e S. Paulo, quanto ao seu comportamento diante da arabinose e xilose. Revelaram os resultados que 472 culturas (58,05%) caracterizaram-se como tipo fermentativo I (acidificando apenas a xilose), 334 amostras (41,08%) per­tencei am ao biotipo II (ausência de fermentação em arabinose e xilose), tendo apenas 7 culturas (0,86%) representando o biotipo III (fermentam a arabino­se e xilose).

Atividade «in vitro» da ampicilina e hetacilina sobre amostras de Salmonella typhi

Os autores, utilizando a técnica da diluição seriada em meio líquido, determinaram as concentrações mínimas inibitórias da ampicüina e da hetacilina em 150 amostras de Salmonella typhi, isoladas em sua totalidade a partir de hemoculturas, no Estado da Guanabara. Obteve-se como média das concentrações bacteriostáticas, 0,545 mcg/ml para ampicilina e 0,720 mcg/ml, no caso da hetacilina.

Inatividade "in vitro" do Hycanthone relativamente a bactérias do gênero Salmonella

Verificaram os autores que, "in vitro", concentrações de "Hycanthone" variáveis de 0,39 a 400 mcg/ml não impediram o desenvolvimento de Salmonella anatum, S. derby, S. typhi e S. typhimurium. Essa concentração pode, sem dúvida, representar apoio à hipótese segundo a qual, na salmonelose de curso prolongado, há carreamento das bactérias pelos helmintos, uma vez que Macêdo & cols. curaram, recentemente, a um só tempo, a esquistossomíase mansônica e a infecção bacteriana de pacientes acometidos pelo citado e intrigante problema da patologia regional brasileira.

Implicações para a saúde pública de amostras de Salmonella com resistência múltipla infecciosa a drogas isoladas de animais

Com a finalidade de verificar se a ocorrência de Salmonelas isoladas de animais, com resistência múltipla infecciosa a drogas, influencia a situação epidemiológica no homem, testaram-se 136 amostras de Salmonella isoladas de suínos normas. Das culturas testadas, 18,3% foram portadoras desse tipo de resistência, em evidente oposição aos dados encontrados entre as amostras de origem humana, em que num total de 70 amostras (67,1%) apresentaram resistência a uma e a várias drogas.

Isolamento de Salmonella typhimurium fermentadora de lactose, em caso autoctone no Rio de Janeiro

Relata-se o primeiro isolamento no Rio de Janeiro, a partir de um caso de gastroenterite infantil, da variante de Salmonella typhimurium, que se tornou prevalente na cidade de São Paulo. O mesmo padrão de resistência múltipla a agentes antibacterianos foi observado, com sensibilidade à gentamicina, colimicina e a associação sulfametaxazol-trimetoprim. Discutem-se características bacteriológicas das amostras de mesmo comportamento, enfatizando-se a necessidade de adaptação das técnicas bacterio­lógicas visando a sua detecção, que provavelmente vem passando desapercebida pelas técnicas tradicionais.