Asymptomatic Plasmodium infection in a residual malaria transmission area in the Atlantic Forest region: Implications for elimination.

INTRODUCTION: Elimination of malaria in areas of interrupted transmission warrants careful case assessment to avoid the reintroduction of this disease. Occasional malaria cases are reported among visitors of the Atlantic Forest area of Brazil, while data on residents of this area are scarce. METHODS: A sectional study was carried out to examine 324 individuals living in a municipality where autochthonous cases were detected. RESULTS: Asymptomatic Plasmodium infections were detected in 2.8% of the individuals by polymerase chain reaction (PCR), with one case of P. falciparum (0.3%), two cases of P. vivax (0.6%), and six cases of P. malariae (1.9%). The thick blood smears were negative in all individuals. Serological tests performed in 314 subjects were reactive in 11.1%, with 3.5% for P. falciparum and 7.7% for P. vivax. A subsample of 42 reactive individuals for any Plasmodium species showed P. malariae in 30.9% of specimens. Individuals who entered the Atlantic Forest region were 2.7 times more likely to exhibit reactive serology for P. vivax compared with individuals who did not enter this region (p<0.05). Children <15 years had a higher chance of reactive serology for P. falciparum and P. vivax than individuals ≥15 years of age (p<0.05). Individuals living in the Paraiso district had a higher chance of reactive serology for P. vivax compared to other districts (p<0.05). No associations were found between sex, past exposure to malaria, or serological response to antibodies of any Plasmodium species. CONCLUSIONS: The implications of these results for the elimination of malaria were discussed.

Asymptomatic plasmodium infection in a residual malaria transmission area in the atlantic forest region: implications for elimination

Abstract INTRODUCTION: Elimination of malaria in areas of interrupted transmission warrants careful case assessment to avoid the reintroduction of this disease. Occasional malaria cases are reported among visitors of the Atlantic Forest area of Brazil, while data on residents of this area are scarce. METHODS: A sectional study was carried out to examine 324 individuals living in a municipality where autochthonous cases were detected. RESULTS: Asymptomatic Plasmodium infections were detected in 2.8% of the individuals by polymerase chain reaction (PCR), with one case of P. falciparum (0.3%), two cases of P. vivax (0.6%), and six cases of P. malariae (1.9%). The thick blood smears were negative in all individuals. Serological tests performed in 314 subjects were reactive in 11.1%, with 3.5% for P. falciparum and 7.7% for P. vivax. A subsample of 42 reactive individuals for any Plasmodium species showed P. malariae in 30.9% of specimens. Individuals who entered the Atlantic Forest region were 2.7 times more likely to exhibit reactive serology for P. vivax compared with individuals who did not enter this region (p<0.05). Children <15 years had a higher chance of reactive serology for P. falciparum and P. vivax than individuals ≥15 years of age (p<0.05). Individuals living in the Paraiso district had a higher chance of reactive serology for P. vivax compared to other districts (p<0.05). No associations were found between sex, past exposure to malaria, or serological response to antibodies of any Plasmodium species. CONCLUSIONS: The implications of these results for the elimination of malaria were discussed.

Genetic diversity of Cryptosporidium identified in clinical samples from cities in Brazil and Argentina.

The identification and characterisation of Cryptosporidium genotypes and subtypes are fundamental to the study of cryptosporidiosis epidemiology, aiding in prevention and control strategies. The objective was to determine the genetic diversity of Cryptosporidium in samples obtained from hospitals of Rio de Janeiro, Brazil, and Buenos Aires, Argentina. Samples were analysed by microscopy and TaqMan polymerase chain reaction (PCR) assays for Cryptosporidium detection, genotyped by nested-PCR-restriction fragment length polymorphism (RFLP) analysis of the 18S rRNA gene and subtyped by DNA sequencing of the gp60 gene. Among the 89 samples from Rio de Janeiro, Cryptosporidium spp were detected in 26 by microscopy/TaqMan PCR. In samples from Buenos Aires,Cryptosporidium was diagnosed in 15 patients of the 132 studied. The TaqMan PCR and the nested-PCR-RFLP detected Cryptosporidium parvum, Cryptosporidium hominis, and co-infections of both species. In Brazilian samples, the subtypes IbA10G2 and IIcA5G3 were observed. The subtypes found in Argentinean samples were IbA10G2, IaA10G1R4, IaA11G1R4, and IeA11G3T3, and mixed subtypes of Ia and IIa families were detected in the co-infections. C. hominis was the species more frequently detected, and subtype family Ib was reported in both countries. Subtype diversity was higher in Buenos Aires than in Rio de Janeiro and two new subtypes were described for the first time.

Genetic diversity of Cryptosporidium identified in clinical samples from cities in Brazil and Argentina

The identification and characterisation of Cryptosporidiumgenotypes and subtypes are fundamental to the study of cryptosporidiosis epidemiology, aiding in prevention and control strategies. The objective was to determine the genetic diversity ofCryptosporidium in samples obtained from hospitals of Rio de Janeiro, Brazil, and Buenos Aires, Argentina. Samples were analysed by microscopy and TaqMan polymerase chain reaction (PCR) assays forCryptosporidium detection, genotyped by nested-PCR-restriction fragment length polymorphism (RFLP) analysis of the 18S rRNA gene and subtyped by DNA sequencing of the gp60 gene. Among the 89 samples from Rio de Janeiro, Cryptosporidium spp were detected in 26 by microscopy/TaqMan PCR. In samples from Buenos Aires,Cryptosporidium was diagnosed in 15 patients of the 132 studied. The TaqMan PCR and the nested-PCR-RFLP detected Cryptosporidium parvum, Cryptosporidium hominis, and co-infections of both species. In Brazilian samples, the subtypes IbA10G2 and IIcA5G3 were observed. The subtypes found in Argentinean samples were IbA10G2, IaA10G1R4, IaA11G1R4, and IeA11G3T3, and mixed subtypes of Ia and IIa families were detected in the co-infections. C. hominis was the species more frequently detected, and subtype family Ib was reported in both countries. Subtype diversity was higher in Buenos Aires than in Rio de Janeiro and two new subtypes were described for the first time.

Detection and differentiation of Cryptosporidium by real-time polymerase chain reaction in stool samples from patients in Rio de Janeiro, Brazil.

This study reports the first genetic characterisation of Cryptosporidium isolates in Brazil using real-time polymerase chain reaction (RT-PCR). A total of 1,197 faecal specimens from children and 10 specimens from human immunodeficiency virus-infected patients were collected between 1999-2010 and screened using microscopy. Forty-eight Cryptosporidium oocyst-positive isolates were identified and analysed using a generic TaqMan assay targeting the 18S rRNA to detect Cryptosporidium species and two other TaqMan assays to identify Cryptosporidium hominis and Cryptosporidium parvum. The 18S rRNA assay detected Cryptosporidium species in all 48 of the stool specimens. The C. parvum TaqMan assay correctly identified five/48 stool samples, while 37/48 stool specimens were correctly amplified in the C. hominis TaqMan assay. The results obtained in this study support previous findings showing that C. hominis infections are more prevalent than C. parvum infections in Brazil and they demonstrate that the TaqMan RT-PCR procedure is a simple, fast and valuable tool for the detection and differentiation of Cryptosporidium species.

Detection and differentiation of Cryptosporidium by real-time polymerase chain reaction in stool samples from patients in Rio de Janeiro, Brazil

This study reports the first genetic characterisation of Cryptosporidium isolates in Brazil using real-time polymerase chain reaction (RT-PCR). A total of 1,197 faecal specimens from children and 10 specimens from human immunodeficiency virus-infected patients were collected between 1999-2010 and screened using microscopy. Forty-eight Cryptosporidium oocyst-positive isolates were identified and analysed using a generic TaqMan assay targeting the 18S rRNA to detect Cryptosporidium species and two other TaqMan assays to identify Cryptosporidium hominis and Cryptosporidium parvum. The 18S rRNA assay detected Cryptosporidium species in all 48 of the stool specimens. The C. parvum TaqMan assay correctly identified five/48 stool samples, while 37/48 stool specimens were correctly amplified in the C. hominis TaqMan assay. The results obtained in this study support previous findings showing that C. hominis infections are more prevalent than C. parvum infections in Brazil and they demonstrate that the TaqMan RT-PCR procedure is a simple, fast and valuable tool for the detection and differentiation of Cryptosporidium species.

[Delayed diagnosis of malaria in a dengue endemic area in the Brazilian extra-Amazon: recent experience of a malaria surveillance unit in state of Rio de Janeiro]. / Diagnóstico tardio de malária em área endêmica de dengue na extra-Amazônia Brasileira: experiência recente de uma unidade sentinela no estado do Rio de Janeiro.

INTRODUCTION: The mortality of malaria in the extra-Amazon region is about 80 times higher than in the Amazon region, where malaria is concentrated (99.8% of cases). In areas of dengue transmission, delay in the diagnosis and treatment of malaria in patients with fever who reside in areas of malaria transmission can be due to the confusion between the clinical diagnoses of both diseases by nonspecialist doctors, among other factors. This work presents some of the consequences of delayed diagnosis in three patients with malaria by Plasmodium falciparum, P. malariae and P. vivax, who, after following the usual route for Dengue treatment, sought our institution, where they were correctly diagnosed and adequately treated. METHODS: Description of three cases of malaria with delayed diagnosed malaria referred to the Outpatient Clinic for Acute Febrile Diseases, IPEC/FIOCRUZ-RJ, between 2007 and 2008. RESULTS: A Brazilian from Mozambique, primo-infected with P. falciparum was diagnosed with malaria six days after the onset of fever and died of cerebral malaria and shock. Another patient with P.malariae malaria presented a severe and prolonged course, but was cured after specific treatment. A third patient, with delayed diagnosis of P. vivax malaria, acquired it in the Atlantic Forest region in the State of Rio. CONCLUSIONS: Health professionals from non-endemic areas for malaria should be trained to optimize the surveillance and early treatment of malaria and prevent morbid and fatal outcomes. An investigation of outbreaks of autochthonous malaria in the State of Rio de Janeiro is suggested.

Diagnóstico tardio de malária em área endêmica de dengue na extra-Amazônia Brasileira: experiência recente de uma unidade sentinela no estado do Rio de Janeiro / Delayed diagnosis of malaria in a dengue endemic area in the Brazilian extra-Amazon: recent experience of a malaria surveillance unit in state of Rio de Janeiro

INTRODUÇÃO: A letalidade da malária na região extra-amazônica é cerca de 80 vezes maior do que na Amazônia, que concentra 99,8 por cento dos casos do país. Em áreas de transmissão de dengue, como o Rio de Janeiro, o atraso no diagnóstico e tratamento da malária dos pacientes com febre, provenientes de áreas endêmicas de malária, pode ser, entre outros fatores, devido à confusão entre o diagnóstico das duas doenças pelos generalistas da rede de assistência médica. Neste trabalho, apresentamos as consequências do atraso diagnóstico em três pacientes com malária por Plasmodium falciparum; P. malariae e P. vivax, que, após o périplo habitual para tratamento de dengue, procuraram a nossa instituição onde foram corretamente diagnosticados e submetidos aos tratamentos adequados. MÉTODOS: Descrição de três casos de malária diagnosticada tardiamente e encaminhados ao IPEC/ FIOCRUZ, entre os anos de 2007 e 2008. RESULTADOS: uma brasileira proveniente de Moçambique, primo-infectada por P. falciparum, com malária diagnosticada após 6 dias do início da febre, morreu com malária cerebral e choque. Outro paciente com malária por P. malariae teve um curso grave e prolongado, mas ficou curado após o tratamento específico. A terceira paciente diagnosticada tardiamente apresentou malária por P. vivax adquirida na região de Mata Atlântica no Estado do Rio. CONCLUSÕES: Os profissionais de saúde do Rio devem ser treinados para aperfeiçoar a vigilância e o tratamento oportuno da malária e evitar desfechos mórbidos e fatais. Sugere-se que uma investigação de focos de malária autóctone em áreas de mata no estado seja realizada.

INTRODUCTION: The mortality of malaria in the extra-Amazon region is about 80 times higher than in the Amazon region, where malaria is concentrated (99.8 percent of cases). In areas of dengue transmission, delay in the diagnosis and treatment of malaria in patients with fever who reside in areas of malaria transmission can be due to the confusion between the clinical diagnoses of both diseases by nonspecialist doctors, among other factors. This work presents some of the consequences of delayed diagnosis in three patients with malaria by Plasmodium falciparum, P. malariae and P. vivax, who, after following the usual route for Dengue treatment, sought our institution, where they were correctly diagnosed and adequately treated. METHODS: Description of three cases of malaria with delayed diagnosed malaria referred to the Outpatient Clinic for Acute Febrile Diseases, IPEC/FIOCRUZ-RJ, between 2007 and 2008. RESULTS: A Brazilian from Mozambique, primo-infected with P. falciparum was diagnosed with malaria six days after the onset of fever and died of cerebral malaria and shock. Another patient with P.malariae malaria presented a severe and prolonged course, but was cured after specific treatment. A third patient, with delayed diagnosis of P. vivax malaria, acquired it in the Atlantic Forest region in the State of Rio. CONCLUSIONS: Health professionals from non-endemic areas for malaria should be trained to optimize the surveillance and early treatment of malaria and prevent morbid and fatal outcomes. An investigation of outbreaks of autochthonous malaria in the State of Rio de Janeiro is suggested.

Endoparasitos em cobaias (Cavia porcellus) (Mammalia, Rodentia, Caviidae) provenientes de biotérios de criação e experimentação do município do Rio de Janeiro, Brasil / Endoparasites in guinea pigs (Cavia porcellus) (Mammalia, Rodentia, Caviidae) from breeding and experimentation animal housing of the municipality of Rio de Janeiro, Brazil

Foi realizado um levantamento sobre a prevalência e intensidade de infecção de endoparasitos em cobaias convencionais de linhagem Short Hair provenientes de biotérios de criação (A) e experimentação (B) do município do Rio de Janeiro, Brasil, avaliando-se a eficácia das medidas de prevenção entre eles. Para a realização do estudo, utilizou-se exame direto de mucosa e do conteúdo intestinal, a técnica de tricromo de WHEATLEY e exames coproparasitológicos pelas técnicas de Ritchie e Kinyoun. Os parasitos encontrados através da técnica de exame direto da mucosa e do conteúdo intestinal e de tricromo de WHEATLEY com as respectivas prevalências foram: Balantidium sp. (78 por cento), Cyathodinium sp. (68 por cento), Eimeria caviae (38 por cento), Paraspidodera uncinata (34 por cento) e Giardia muris (24 por cento). Nos exames de fezes realizados pelas técnicas de Ritchie e Kinyoun, foram encontrados os seguintes parasitos com as respectivas prevalências no biotério A: E. caviae (74 por cento), Balantidium sp. (68 por cento), Cyathodinium sp. (68 por cento) e Cryptosporidium sp. (5 por cento). No biotério B, observou-se: E. caviae (58 por cento), Balantidium sp. (42 por cento), Cyathodinium sp. (25 por cento) e G. muris (8 por cento). A alta prevalência de endoparasitos nos biotérios sugere a necessidade de se rever a eficácia das barreiras sanitárias adotadas.

This paper discusses the prevalence and intensity of infection of endoparasites in conventionally maintained Short Hair guinea pigs colonies from a breeding (A) and an experimental (B) facilities in Rio de Janeiro, Brazil. It also evaluates the efficacy of the methods of prevention adopted by both facilities. The search of parasites was performed by direct examination of intestinal mucosa and its contents, WHEATLEY's trichrome method and coproparasitological examinations by Ritchie's and Kinyoun's techniques. The prevalences of endoparasites found throught direct examination of intestinal mucosa and its contents were: Balantidium sp. (78 percent), Cyathodinium sp. (68 percent), Eimeria caviae (38 percent), Paraspidodera uncinata (34 percent) and Giardia muris (24 percent). The prevalences of endoparasites found throught Ritchie's and Kinyoun's techniques in facility A were: E. caviae (74 percent), Balantidium sp. (68 percent), Cyathodinium sp. (68 percent) and Cryptosporidium sp. (5 percent). In facility B: E. caviae (58 percent), Balantidium sp. (42 percent), Cyathodinium sp. (25 percent) and G. muris (8 percent) were found. The high prevalences in both facilities suggest that the efficacy of the sanitary barriers adopted in the prevention of parasitic infections should be revised.

Caracterização morfométrica e avaliação da heterogeneidade molecular de Cryptosporidium sp de grupos populacionais no município do Rio de Janeiro / Morphometric characterization and evaluation of the molecular heterogeneidade in isolated physicians of Cryptosporidium sp of population groups in the city of Rio De Janeiro

Cryptosporidium sp. é um protozoário amplamente disperso em numerosas espécies de hospedeiros por todos os continentes. Estudos de epidemiologia molecular têm mostrado que múltiplos genótipos podem circular numa mesma área geográfica. Tais estudos realçam também, que C. hominis e C. parvum são as espécies mais comuns na infecção humana do que as espécies zoonóticas. Entretanto, todas podem infectar tanto indivíduos imunocompetentes quanto imunodeficientes. Nesse trabalho foram analisados isolados clínicos de Cryptosporidium sp. de 14 crianças (0-10 anos) que apresentaram diarréia com suspeita clínica de Rotavírus (grupo 1), 5 crianças (0-4 anos) de uma creche (grupo 2) e 7 pacientes adultos, HIV+ do Hospital Universitário Pedro Ernesto (grupo 3). Objetivamos caracterizar esses isolados circulantes no município do Rio de Janeiro empregando a análise morfométrica (maior e menor diâmetro, perímetro, área e índice morfométrico) de oocistos corados pelo método de Kinyoun a frio, por meio de software para análise de imagens (Image - Pro Plus - Media Cybernetics) e genotipagem, mediante a reação de PCR e da análise por RFLP dos produtos obtidos da nested-PCR. A extração de DNA genômico foi realizada de fezes totais, utilizando solução de tiocianato de guanidina, pérolas de vidro, e polivinilpirrolidona para remoção parcial de inibidores fecais. Os genes alvos para amplificação foram o COWP e o 18S rDNA. Os produtos da nested-PCR para o gene COWP foram digeridos com a enzima RsaI e os de 18S rDNA, com SspI. Foram encontradas diferenças morfométricas significativas em todos os caracteres (p<0,05), independentes dos grupos. Com base nos caracteres mais utilizados (maior e menor diâmetros), os valores encontrados correspondem a C. hominis e C. parvum, não sendo possível distinção morfométrica entre eles. O índice morfométrico em torno de 1,0 não diferiu entre os isolados. Das 26 amostras analisadas molecularmente, 22 apresentaram perfil característico do genótipo de C. hominis, 2 com perfis compatíveis com genótipo zoonótico de C. parvum e 2 com padrão misto, sendo a diferenciação possibilitada pelas análises de RFLP do gene COWP. Conclui-se que no município do Rio de Janeiro há mais de um genótipo de Cryptosporidium circulante, C. hominis é o mais prevalente, e a caracterização desse organismo deve ser realizada pela genotipagem, devido à semelhança biométrica entre os genótipos.

Ocorrência de Cryptosporidium sp em amostras fecais de crianças, menores de 10 anos de idade, com indicaçäo clínica de Rotavírus / Occurrence of Cryptosporidium sp in fecal samples of children less than 10 years old with clinical indication of Rotavirus

A partir da indicaçäo clínica de Rotavírus em fezes de 485 crianças, investigou-se a presença de oocistos de Cryptosporidium sp. Näo houve diferenças significativas entre a positividade de Cryptosporidium sp e rotavírus com a consistência das fezes. Cryptosporidium sp deve ser incluído na investigaçäo diagnóstica dos quadros diarréicos em crianças

Diagnóstico morfológico da cyclospora cayetanensis: um parasito emergente / Morphological diagnosis of cyclospora cayetanensis: an amergent pathogen

Criptosporidiose e isosporose säo causas bem estabelecidas de diarréia crônica em pacientes com a Sindrome de Imunodeficiência Adquirida (SIDA) e outros estados de imunodeficiência, sendo algumas vezes fatal. Recentemente, um novo coccideo morfologicamente semelhantes ao Cryptosporidium, foi descoberto em fezes de indivíduos imunocompetentes e imunocomprometidos com diarréia. Visto que o quadro clínico da infecçäo pela cyclospora cayetanenesis é similar ao da criptosporidiose, este novo organismo poderia facilmente ser confundido com o Cryptosporidium. No entanto neste artigo apresentamos características morfológicas e de coloraçäo da Cyclospora cayetanensis que nos permitem distingui-la do Cryptosporidium