Treatment of T. cruzi infected human platelet concentrates with aminomethyltrimethyl psoralen (AMT) and ultravioleta (UV-A) light: preliminary results

The present measures adopted to prevent transfusion-associated Chagas' disease include screening of blood donors. and/or the inactivation of T. cruzi in collected blood using gentian violet (GV) as a trypanocidal agent. In this study, we investigated the efficacy of the combined use of AMT and UV-A in inactirating T. cruzi in infected human platelet cuncentrates. Human platelet concentrates were infected with T. cruzi (2x10/ml) of the Y strain transfered to PL 269 (Fenwal Laboratories) containers and treated with GV (250řg,/ml). and ascorbic acid (1 mg/ml); GV. ascorbic acid and UV-A; GV and UV-A; AMT (40/tG/ml) and ascorbic acid; AMT, ascorbic acid and UV-A; AMT and UV-A; UV-A alone; and untreated (control). All UV-A treated platelet concentrates were exposed to UV-A doses of 24, 92, 184, 276, 368 and 644 kj/m². and the microscopical research of active T. cruzi was performed, using the microhematocrit technique, 1, 6 and 24 hours after each treatment. A high number of active forms of T. cruzi was observed in all condictions, except when GV was used as the trypanocidal agent, providing evidence of the failure of AMT and UV-A in inactivating T cruzi in infected human platelet concentrates.

Pesquisa de bactérias patogênicas em leite pasteurizado tipo C comercializado na cidade do Recife, Pernambuco, Brasil

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.

Peritonite bacteriana espontânea

A peritonite bacteriana espontânea ocorre em 30% dos cirróticos com ascite e, neste grupo, apresenta altas taxas de morbidade e mortalidade. Os fatores predisponentes incluem a diminuição da defesa imunológica encontrada no homem nas fases avançadas da cirrose, o supercrescimento da flora intestinal e a translocação bacteriana da luz dos intestinos aos linfonodos mesentéricos. As manifestações clínicas variam de graves a leves ou ausentes, sendo sempre necessária a análise do líquido ascítico. O diagnóstico de peritonite bacteriana espontânea se faz pela contagem de neutrófilos > 250/mm³ no líquido ascítico associado ou não ao crescimento de bactéria na cultura. As enterobactérias predominam como causa da infecção, sendo a Echerichia coli a bactéria mais freqüentemente isolada. O diagnóstico precoce e o tratamento adequado provocaram a queda das taxas de mortalidade nas duas últimas décadas. O uso endovenoso de cefalosporinas de terceira geração mostra-se eficaz em 70% a 95% dos casos. A recorrência de peritonite bacteriana espontânea é comum e pode ser prevenida com norfloxacina oral, de uso contínuo. O surgimento de resistência bacteriana tem estimulado a procura de novas opções para a profilaxia da peritonite bacteriana espontânea; os probióticos constituem nova abordagem promissora, mas que necessita melhor avaliação. Recomenda-se a profilaxia primária de curta duração aos cirróticos com ascite que apresentem episódio de hemorragia digestiva alta.

Experimental schistosomiasis in the Common Marmoset Callithrix jacchus

In order to evaluate Callithrix jacchus as an animal model for mansoni schistosomiasis, a group of 10 male animals were once percutaneously exposed to 250 cercariae of the Schistosoma mansoni SLM (São Lourenço da Mata) strain. Animals were periodically bled for measuring serum level of enzymes and proteins and for blood cell counting. When comparing pre-infection to post-infection values, a significant increase was found for alkaline phosphatase at 15 to 120 days p.i., differential counts of eosinophil at 45 and 60 days, and total protein and global eosinophil counts at 120 days. No Schistosoma mansoni eggs were found in stools. Adult worms of small size were recovered from five animals. At day 120, the number of Schistosoma mansoni eggs/g of tissue was 0-289.7 (liver), 0-30.1 (large intestine) and 0-171.4 (small intestine). These findings lead us to classify Callithrix jacchus as a non-permissive host to the SLM strain of Schistosoma mansoni.

Soroprevalência da hepatite C em pacientes hemodialisados

Os objetivos deste estudo foram estimar a soroprevalência da infecção pelo vírus da hepatite C em um centro de diálise da grande Recife; e associar a soropositividade para o vírus da hepatite C em relação a alguns fatores de risco. Foram analisados 250 pacientes com idade variando de 17 a 92 anos e de ambos os sexos. Dados epidemiológicos desses pacientes foram obtidos para a determinação dos fatores de riscos para esta infecção. A pesquisa de anticorpos anti-HCV foi realizada pelo ELISA de 4ª geração. Foi observado que em relação aos fatores de riscos, como o tempo de hemodiálise, número e período das transfusões de hemocomponentes, foi encontrada uma associação estatisticamente significante (p< 0,05). A prevalência encontrada foi baixa (8,4%) em relação a outros estudos do Brasil. Entretanto, seriam necessários mais estudos em outros centros a fim de estimar a real prevalência para infecção pelo vírus da hepatite C em pacientes submetidos a hemodiálise em Pernambuco.

Suscetibilidade a antifúngicos de variedades de Cryptococcus neoformans isoladas de pacientes em hospital universitário

Este trabalho identificou variedades de Cryptococcus neoformans e avaliou a suscetibilidade a antifúngicos pelo protocolo M27-A2 do National Committee for Clinical Laboratory Standards em isolados de 35 pacientes do Hospital Escola da Universidade Federal do Triângulo Mineiro. A variedade gatti foi identificada em 11.4% (nº = 4) dos casos. A concentração inibitória mínima (mg/ml) dos isolados de Cryptococcus neoformans neoformans variou de 0,062 - 2,000 (anfotericina B), 0,250 - 8,000 (fluconazol), 0,062 - 1,000 (itraconazol) e 0,125 - 1,000 (cetoconazol). A variedade gattii apresentou concentração inibitória mínima de 0,125 - 2,000 (anfotericina B), 0,250 - 16,00 (fluconazol), 0,062 - 1,000 (itraconazol) e 0,125 - 4,000 (cetoconazol). Detectaram-se 2 isolados resistentes ao itraconazol e 2 a anfotericina B (1 isolado de cada variedade por antifúngico). Os dados mostram a importância da determinação da variedade e da concentração inibitória mínima de isolados de Cryptococcus neoformans para monitorar o desenvolvimento de resistência e possibilitar uma terapia mais adequada na criptococose.

Gender influence on treatment of chronic hepatitis C genotype 1

INTRODUCTION: Although various studies have been published regarding the treatment of chronic hepatitis C (CHC) with peginterferon (Peg-IFN) and ribavirin, little is known regarding the real impact of gender on the characteristics that influence the effectiveness and safety of antiviral treatment for CHC patients. The objective of this study was to evaluate the influence of gender on HCV treatment outcomes. METHODS: A retrospective analytical study was conducted among selected carriers of CHC genotype 1, who were treated with Peg-IFN α-2b at a dose of 1.5 μg/kg or Peg-IFN α-2a at a dose of 180 μg/week plus a ribavirin dose of 1,000-1,250 mg/day, according to weight, between 2001 and 2007. RESULTS: Among 181 patients undergoing treatment, the mean age was 46.4 ± 11.0 years and 46% were women. At baseline, 32% of the patients had advanced fibrosis (F3-F4 Scheuer), and 83% of the subjects had viral load > 400,000 IU/ml, without significant difference between the genders (p = 0.428 and p = 0.452, respectively). When compared with men, women had higher incidence of many adverse events such as anemia (p < 0.001) and higher need for dose reduction, for both Peg-IFN (p = 0.004) and ribavirin (p = 0.006). However, the rate of sustained virological response (SVR) did not differ between the genders: 45% (female) vs 41% (male); p=0.464. CONCLUSIONS: This study suggests that women and men react differently to combined therapy, especially in relation to the incidence of adverse events and the need for dose modification. Nevertheless, these differences do not influence the SVR rate.

Production of anti-Cryptosporidium polyclonal antibodies and standardization of direct immunofluorescence for detecting oocysts in water

INTRODUCTION: The production of anti-Cryptosporidium polyclonal antibodies and its use in direct immunofluorescence assays to determine the presence of Cryptosporidium in water are described in the present work. METHODS: Two rabbits were immunized with soluble and particulate antigens from purified Cryptosporidium oocysts. The sera produced were prepared for immunoglobulin G extraction, which were then purified and conjugated with fluorescein isothiocyanate (FITC). Slides containing known amounts of oocysts were prepared to determine the sensitivity of the technique. To test the specificity, slides containing Giardia duodenalis cysts were prepared. RESULTS: The conjugate was successfully used in water samples experimentally contaminated with Cryptosporidium oocysts, and it was possible to detect up to five oocysts/spot, corresponding to contamination of 250 oocysts/mL. CONCLUSIONS: The three immunizations performed in the rabbits were enough to produce antibodies against Cryptosporidium, the standard direct immunofluorescence assay permitted the detection of five oocysts in 20% of the samples, and no cross-reaction with Giardia duodenalis cysts occurred.

Detection of Dientamoeba fragilis in patients with HIV/AIDS by using a simplified iron hematoxylin technique

INTRODUCTION: Studies strongly indicate Dientamoeba fragilis as one of the causes of diarrhea in human immunodeficiency virus (HIV) patients. METHODS: The objective of the present study was to evaluate the prevalence of D. fragilis associated with the causes of diarrhea in 82 HIV/ AIDS patients hospitalized at the Instituto de Infectologia Emílio Ribas from September 2006 to November 2008. RESULTS: In total, 105 samples were collected from 82 patients. Unprotected sex was the most frequent cause of HIV infection (46.3%), followed by the use of injectable or non-injectable drugs (14.6%). Patients presented with viral loads of 49-750,000 copies/ mL (average: 73,849 ± 124,850 copies/mL) and CD4 counts ranging of 2-1,306 cells/mm³ (average: 159 ± 250 cells/mm³). On an average, the odds of obtaining a positive result by using the other techniques (Hoffman, Pons and Janer or Lutz; Ritchie) were 2.7 times higher than the chance of obtaining a positive result by using the simplified iron hematoxylin method. Significant differences were found between the methods (p = 0.003). CONCLUSIONS: The other techniques can detect a significantly greater amount of parasites than the simplified iron hematoxylin method, especially with respect to Isospora belli, Cryptosporidium sp., Schistosoma mansoni, and Strongyloides stercoralis, which were not detected using hematoxylin. Endolimax nana and D. fragilis were detected more frequently on using hematoxylin, and the only parasite not found by the other methods was D. fragilis.

Increased detection of schistosomiasis with Kato-Katz and SWAP-IgG-ELISA in a Northeastern Brazil low-intensity transmission area

INTRODUCTION: The laboratory diagnosis of schistosomiasis is based mainly on the detection of parasite eggs in stool samples through the Kato-Katz (KK) technique, reading one slide by test. However, a widely known limitation of parasitological methods is reduced sensitivity, particularly in low endemic areas. METHODS: To increase sensitivity, we conducted further slide readings from the same stool sample using the parasitological method associated with a serological test. We used the KK method (three slides) and the IgG anti-Schistosoma mansoni-enzyme-linked immunosorbent assay (ELISA) technique to diagnose schistosomiasis in low endemic areas in the Brazilian State of Ceará. Fecal samples and sera from 250 individuals were analyzed. RESULTS: Sixteen percent and 47.2% of samples were positive in parasitological tests and serological tests, respectively. Parasitological methods showed that 32 (80%) individuals tested positive on the first slide, 6 (15%) on the second slide, and 2 (5%) on the third. The performance of the ELISA test in the diagnosis, using the KK method as diagnostic reference, showed a negative predictive value of 100%, with specificity and positive predictive values of 62.8% and 33.9%, respectively. CONCLUSIONS: In this study, the increase from one to three slides analyzed per sample using the KK technique was shown to be a useful procedure for increasing the diagnostic sensitivity of this technique.

Snakebite cases in the municipalities of the State of Paraiba, Brazil

Introduction This study investigated the epidemiological and clinical profile of snakebite cases reported from 2007 to 2012 in the municipalities of the Curimataú region, State of Paraíba, in northeastern Brazil. Methods Data were collected from the Health Department of the State of Paraíba using the Injury Notification Information System data banks of the Health Ministry. Results A total of 304 snakebite cases were studied. The cases occurred most frequently from April to June. The genera Bothrops, Crotalus, and Micrurus were responsible for 74.6%, 6.2%, and 1.3% of cases, respectively. Snakebite cases predominated in males living in rural areas and between 10 and 19 years old. The highest incidence of bites occurred on the feet. The majority of the victims received medical assistance within 1 to 3h after being bitten. With regard to severity, 48% of the cases were classified as mild, 26% as moderate, and 2.6% as severe. Successful cures predominated, and no deaths were reported. The average antivenom ampoule dose used for the treatment in some snakebite cases was lower than that recommended by the Health Ministry. Conclusions Although our results show that Paraíba has a good level of medical care, there are serious deficiencies in recording snakebite information. These data indicate the need to improve the recording process for snakebite cases. Further training for health professionals seems to be necessary to optimize their skills in treating snakebite victims.

A malacological survey in the Manso Power Plant, State of Mato Grosso, Brazil: new records of freshwater snails, including transmitters of schistosomiasis and exotic species

Introduction Schistosomiasis is a parasitic disease of public health concern in Brazil, and the construction of hydroelectric dams, in addition to increasing permanent human settlement and tourism, has created conditions suitable for the establishment of mollusks that can transmit schistosomiasis. Such areas require a number of actions to prevent the establishment of schistosomiasis. This paper reports on a freshwater malacological survey carried out in the geographical area of the Manso Power Plant. Methods Mollusks were collected in 18 municipalities in the State of Mato Grosso between February 2002 and February 2004 (qualitative study) and from April 2009 to February 2011 (quantitative study). Results Thirty-one species of mollusks were collected, including newly recorded species (Antillorbis nordestensis and Burnupia ingae). In addition, the geographic distributions of known species, including Biomphalaria straminea, a snail vector of Schistosoma mansoni, were expanded. A total of 4,507 specimens were collected in the APM Manso reservoir (Usina Hidrelétrica de Aproveitamento Múltiplo de Manso) during the quantitative study, and Biomphalaria amazonica was found in six of the 10 localities analyzed. The Afroasiatic species Melanoides tuberculata, introduced after February 2009, was the dominant species (relative abundance 94.96%). Conclusions The study area is epidemiologically important due to the occurrence of B. straminea and B. amazonica, which are vectors of schistosomiasis, and M. tuberculata, a snail host of Centrocestus formosanus, which is responsible for centrocestiasis transmission. Observations of M. tuberculata and the exotic freshwater clams Corbicula fluminea and Corbicula largillierti raise concerns about biodiversity.

Leptospirosis in animals and human contacts in Egypt: broad range surveillance

INTRODUCTION: Leptospirosis is a re-emerging zoonotic disease of humans and animals worldwide. The disease is caused by pathogenic species of the genus Leptospira. These organisms are maintained in nature via chronic renal infection of carrier animals, which excrete the organisms in their urine. Humans become infected through direct or indirect exposure to infected animals and their urine or through contact with contaminated water and soil. This study was conducted to investigate Leptospira infections as a re-emerging zoonosis that has been neglected in Egypt. METHODS: Samples from 1,250 animals (270 rats, 168 dogs, 625 cows, 26 buffaloes, 99 sheep, 14 horses, 26 donkeys and 22 camels), 175 human contacts and 45 water sources were collected from different governorates in Egypt. The samples were collected from different body sites and prepared for culture, PCR and the microscopic agglutination test (MAT). RESULTS: The isolation rates of Leptospira serovars were 6.9%, 11.3% and 1.1% for rats, dogs and cows, respectively, whereas the PCR results revealed respective detection rates of 24%, 11.3% and 1.1% for rats, dogs and cows. Neither the other examined animal species nor humans yielded positive results via these two techniques. Only six Leptospira serovars (Icterohaemorrhagiae, Pomona, Canicola, Grippotyphosa, Celledoni and Pyrogenes) could be isolated from rats, dogs and cows. Moreover, the seroprevalence of leptospiral antibodies among the examined humans determined using MAT was 49.7%. CONCLUSIONS: The obtained results revealed that rats, dogs and cows were the most important animal reservoirs for leptospirosis in Egypt, and the high seroprevalence among human contacts highlights the public health implications of this neglected zoonosis.

Factors affecting Helicobacter pylori eradication using a seven-day triple therapy with a proton pump inhibitor, tinidazole and clarithromycin, in brazilian patients with peptic ulcer

Triple therapy is accepted as the treatment of choice for H. pylori eradication. In industrialized countries, a proton pump inhibitor plus clarithromycin and amoxicillin or nitroimidazole have shown the best results. Our aims were: 1. To study the eradication rate of the association of a proton pump inhibitor plus tinidazole and clarithromycin on H. pylori infection in our population. 2. To determine if previous treatments, gender, age, tobacco, alcohol use, and non-steroidal anti-inflammatory drugs (NSAIDs) change the response to therapy. METHODS: Two hundred patients with peptic ulcer (upper endoscopy) and H. pylori infection (histology and rapid urease test - RUT) were included. A proton pump inhibitor (lansoprazole 30 mg or omeprazole 20 mg), tinidazole 500 mg, and clarithromycin 250 mg were dispensed twice a day for a seven-day period. Eradication was assessed after 10 to 12 weeks of treatment through histology and RUT. RESULTS: The eradication rate of H. pylori per protocol was 65% (128/196 patients). This rate was 53% for previously treated patients, rising to 76% for not previously treated patients, with a statistical difference p<0.01. No significant difference was observed regarding sex, tobacco use, alcohol consumption, and NSAID use, but for elderly patients the difference was p = 0.05. Adherence to treatment was good, and side effects were mild. CONCLUSIONS: A proton pump inhibitor, tinidazole, and clarithromycin bid for seven days resulted in H. pylori eradication in 65% of the patients. Previous treatments were the main cause of treatment failure.

Policultivo de Matrinchã (Brycon sp.) e Jaraqui (Semaprochilodus sp.) em pequenas represas

Foi estudado o crescimento de matrinchã (Brycon sp.) e jaraqui (Semaprochilodus sp.) em sistema de policultivo em duas pequenas represas de igarapés de terra firme, nos arredores de Manaus, com superfícies de água de aproximadamente 6500 e 1500 m2 . As represas foram povoadas com alevinos capturados na natureza numa taxa de esfocagem de aproximadanente 0,4 - 0,5 peixe/m2 para cada espécie. Os peixes foram alimentados com uma ração comercial para aves poedeiras (18,5% proteína bruto, 5,4% extrato etéreo), sendo a quantidade diária inicial de 4% da biomassa dos matrinchãs até atingirem um peso médio de 250 g e, poste lio rmente, de 3% da biomassa dos mesmos. Mensalmente foi feita amostragem dos peixes e da água, que é quimicamente pobre. Em 307 e 364 dias de experimento nas duas represas foram obtidos, respectivamente, 2035 kg e 506 kg de biomassa total de peixes, o que corresponde a uma produção total calculada de 3,7 t/ha/ano e 3,4 t/ ha/ano.