The need for culture in tegumentary leishmaniasis diagnosis in Bolivia: A comparative evaluation of four parasitological techniques using two sampling methods.

Leishmaniases are zoonotic diseases caused by protozoa of the genus Leishmania. In Bolivia, leishmaniasis occurs mainly in the cutaneous form (CL) followed by the mucosal or mucocutaneous form (ML or MCL), grouped as tegumentary leishmaniosis (TL), while cases of visceral leishmaniasis (VL) are rare. The cases of TL are routinely diagnosed by parasitological methods: Direct Parasitological Exam (DPE) and axenic culture, the latter being performed only by specialized laboratories. The aim of the present study was to optimize the parasitological diagnosis of TL in Bolivia, using two sampling methods. Samples from 117 patients with suspected TL, obtained by aspiration (n = 121) and scraping (n = 121) of the edge of the lesion were tested by: direct parasitological exam, culture in TSTB medium, and miniculture and microculture in Schneider's medium. A positive laboratory result by any of the four techniques evaluated using either of the two sampling methods was considered the gold standard. Of the 117 suspected patients included, TL was confirmed in 96 (82 %), corresponding 79 of the confirmed cases (82.3 %) to CL and 16 (16.7 %) to ML. Parasitological techniques specificity was 100 % and their analytical sensitivity was greater with scraping samples in TSTB culture (98 %). Scraping samples in TSTB and miniculture correlated well with the reference (Cohen's kappa coefficient=0.88) and showed good reliability (Cronbach's alpha coefficient ≥0.91). Microculture provided positive results earlier than the other culture methods (mean day 4.5). By day 14, 98 % of positive cultures had been detected. Scraping sampling and miniculture were associated with higher culture contamination (6 % and 17 %, respectively). Bacterial contamination predominated, regardless of the sampling and culture method, while filamentous fungi and mixed contamination were more frequently observed in cultures from scraping samples. In conclusion: (i) scraping samples proved more suitable for the diagnosis of TL as they increased analytical sensitivity, are less traumatic for the patient and are safer for laboratory personnel than aspirates; (ii) culture, mainly in TSBT medium, should be used for the diagnosis of TL due to its high sensitivity (doubling the number of cases diagnosed by DPE) and its low cost compared to other culture media.

Time of incubation of agar-plate culture for the diagnosis of Strongyloides stercoralis infection.

PURPOSE: Agar-plate culture (APC) remains the most sensitive parasitological technique for S. stercoralis diagnosis. Although it was first described three decades ago, the time of incubation of the plates is neither a commonly described feature nor usually standardized. The aim of the study was to analyze the required time to detect S. stercoralis larvae in APC. METHODS: A prospective laboratory-based study including all patients with at least one positive APC was performed. The plates were incubated at room temperature for 7 days. Clinical, analytical and parasitological features including results of the direct visualization of the stool (DV) after formalin-ether concentration and time-to-detection (TTD) of the larvae in APC were recorded. RESULTS: A total of 141 samples from 75 patients had a positive APC. In 49 of them (65.3%) three or more stool samples were processed for direct visualization (DV) and APC. Of these 49 patients, 8 (16.3%) were also diagnosed with DV and 41 (83.7%) were diagnosed only with APC. In 38 samples from 23 (30.7%) patients, the TTD was below 2 days, while in 27 samples from 13 (17.3%) patients, the larvae were detected on the 6th and 7th day. CONCLUSION: Direct visualization failed to detect S. stercoralis in most of the patients that were diagnosed with APC. Incubation periods below 2 and 5 days would miss an important percentage of infections. At least 7 days of incubation of the APC are required to detect presumably low-burden chronic infections in non-endemic countries.

Platynosomum illiciens (Trematoda: Dicrocoeliidae) in a hybrid marmoset (Callithrix sp.) in the Municipality of Seropédica, RJ, Brazil - Case report / Platynosomum illiciens (Trematoda: Dicrocoeliidae) em um sagui híbrido (Callithrix sp.) no município de Seropédica, RJ, Brasil - Relato de caso

Abstract Platynosomum illiciens is a liver trematode encountered infecting mainly felids although it has also been reported in birds and in additional mammalian species, including non-human primates. The current study reports a natural P. illiciens infection primate of the genus Callithrix. The diagnosis was made using a combination of copro-parasitological techniques, morphological evaluation of adult specimens recovered from the liver during necropsy, and molecular analyses. Eggs were brown in color, oval, operculated, and contained a miracidium. Adult specimens recovered during necropsy were measured and showed dimensions compatible with P. illiciens. Molecular characterization of the trematode involved amplification by polymerase chain reaction (PCR), in combination with nucleotide sequencing, of an approximately 900 base pairs fragment corresponding to 18S-ITS1-5.8S ribosomal DNA. Sequenced amplicons showed 100% nucleotide identity with sequences deposited in the GenBank database as derived from specimens of P. illiciens recovered from cats in Malaysia and Brazil. It was concluded that the morphological and molecular analyses presented herein, confirmed the identification of the trematode recovered as P. illiciens.

Resumo Platynosomum illiciens é um trematódeo do fígado, encontrado principalmente em felinos, embora também tenha sido relatado em aves e outras espécies de mamíferos, incluindo primatas não humanos. Este estudo relata um caso de parasitismo natural por P. illiciens em um primata do gênero Callithrix. O diagnóstico foi feito com uma combinação de técnicas coproparasitológicas, avaliação morfológica de espécimes adultos recuperados do fígado durante a necropsia e análise molecular. Os ovos eram de cor marrom, ovais, operculados e continham um miracídio. Espécimes adultos recuperados durante a necropsia foram medidos e mostraram dimensões compatíveis com P. illiciens. A caracterização molecular do trematódeo envolveu a reação em cadeia da polimerase (PCR) e o sequenciamento de um fragmento de, aproximadamente, 900 pares de bases correspondentes ao DNA robosomal 18S-ITS1-5.8S, que apresentou 100% de identidade da sequência de nucleotídeos com as sequências do GenBank depositadas como derivadas de P. illiciens recuperados de gatos na Malásia e no Brasil, respectivamente. Concluiu-se pelas análises morfológicas e moleculares aqui apresentadas, que o trematódeo é da espécie P. illiciens.

Current status of research regarding Blastocystis sp, an enigmatic protist, in Brazil

The present study aimed to evaluate the occurrence of Blastocystis sp. in Brazilian studies over a period of years (2000-2020), as well as point out relevant aspects of this enigmatic organism. We performed a literature search using six sources of international databases. The data were divided into diagnostic by parasitological and molecular techniques, and relevant aspects. After applying the inclusion and exclusion criteria, 52 studies were included in the final analysis. The occurrence of Blastocystis sp. in Brazil ranged from 0.5% to 86.6%, as determined using parasitological techniques. The highest occurrence was in the North (27.3%) and the lowest, in the Midwest region (13.4%). In Brazil, most studies have employed molecular techniques and are concentrated in the Southeast region. The Blastocystis sp. subtype ST3 had the highest average positivity, followed by ST1 and ST2. These findings represent a panorama that reflects the reality of Brazil; thus, we believe that the effectiveness of parasitological diagnosis should be considered with regard to making an appropriate choice of technique for detecting Blastocystis sp. Additionally, we emphasize the importance of further studies in the context of molecular epidemiology with regard to this genus. Blastocystis sp. is not well understood yet, and very little information regarding this genus is available; hence, further research regarding this genus is urgently needed.

Use of Mini-FLOTAC and Fill-FLOTAC for rapidly diagnosing parasitic infections in zoo mammals / Utilização do Mini-FLOTAC e Fill-FLOTAC no diagnóstico rápido de infecções parasitárias de mamíferos em zoológicos

Abstract Animals reared in restricted environments are highly susceptible to gastrointestinal infection by helminths and protozoa and therefore zoos are characterized as being parasite-rich environments. Successful implementation of control programs of these parasites in zoo environment depends upon precise and rapid diagnosing of gastrointestinal infections. The aim of this study was to demonstrate the role of the Mini-FLOTAC technique in combination with Fill-FLOTAC for rapidly diagnosing parasitic infections in zoo mammals. Fecal samples were collected from 70 animals in four different zoos located in central and southern Italy. All the samples were analyzed using Mini-FLOTAC in combination with Fill-FLOTAC. Out of the 70 pooled samples examined, 80% (24/30) were positive for at least one parasite. Among the gastrointestinal nematodes, Strongyles were the most frequent (40%), followed by Trichuris spp. (23.3%), Parascaris spp. (13.3%) and Capillaria spp. (3.3%). Among the protozoa, Blastocystis spp., Giardia spp. and Eimeria spp. were detected in 6.6%, 3.3% and 3.3%, respectively. These results show that Mini-FLOTAC in combination with Fill-FLOTAC can be used, not only for rapidly diagnosing parasitic infections in zoo mammals, but also for monitoring control programs in which large numbers of fecal samples need to be examined rapidly and reliably.

Resumo Animais criados em ambiente restritos são altamente suscetíveis a infecção gastrointestinal por helmintos e protozoários, constituindo os zoológicos em ambientes com alta contaminação por parasitos. O sucesso da implementação de programas de controle contra estes parasitos em zoológicos depende do rápido diagnóstico das infecções por parasitas gastrointestinais. O objetivo deste estudo foi demonstrar o papel da técnica do Mini-FLOTAC em combinação com o Fill-FLOTAC no diagnóstico rápido das infecções parasitárias em mamíferos em zoológicos. Amostras de fezes foram coletadas de 70 animais de quatro diferentes zoológicos no centro e sudoeste da Itália. Todas as amostras foram analisadas pela técnica do Mini-FLOTAC em combinação com o Fill-FLOTAC. Do total de 70 pools de fezes examinadas, 80% (24/30) foram positivas para pelo menos um parasito. Entre os nematoides gastrointestinais a maior frequência foi observada para estrongilídeos (40%), seguida por Trichuris spp. (23,3%), Parascaris spp. (13,3%) e Capillaria spp. (3,3%). Entre os protozoários Blastocystis spp., Giardia spp. e Eimeria spp. foram detectados em 6,6%, 3,3% e 3,3%, respectivamente. Estes resultados demonstram que a técnica do Mini-FLOTAC em combinação com o Fill-FLOTAC pode ser utilizada não somente para o diagnóstico rápido das infecções parasitárias em mamíferos em zoológicos, mas também no monitoramento de programas de controle onde grande número de amostras fecais devem ser examinadas de forma rápida e confiável.

First parasitological, histopathological and molecular characterization of Echinococcus vogeli Rausch and Bernstein, 1972 from Cuniculus paca Linnaeus, 1766 in the Cerrado biome (Mato Grosso do Sul, Brazil).

Polycystic echinococcosis (PE) is caused by Echinococcus vogeli metacestodes (larval stage) in Neotropical countries. E. vogeli is trophically-transmitted between predators bush dogs (Speothos venaticus) and prey pacas (Cuniculus paca). In Brazil, reported PE cases are restricted to the Amazon biome. In this study, metacestodes from a paca hunted in Mato Grosso do Sul state (Cerrado biome) were identified morphological and histopathological techniques and further confirmed by molecular testing (sequencing of cytochrome C oxidase subunit I (cox1) gene) for the first time. Images of the whole liver showed superficial bubble-like hepatic masses. The parasitological analysis revealed large hooks (41.3 ±â€¯1.2 µm length/12.8 ±â€¯0.8 µm width) and small hooks (33.0 ±â€¯1.5 µm length/11.1 ±â€¯1.2 µm width), consistent with E. vogeli. Microscopically, the liver showed protoscoleces, a thick laminated layer, fibrosis, and inflammatory infiltrate in the adventitial layer. The DNA sequencing confirmed E. vogeli with 99% homology with sequences deposited in the GenBank. In addition, this finding greatly extends the geographic range of animal polycystic echinococcosis into the Cerrado. It is likely to occur in new biomes, where bush dogs and pacas share a given area in a trophic relationship.

Técnicas de citologia aspirativa, biópsia e citobloco de medula óssea para identificação e determinação de intensidade parasitária na leishmaniose visceral canina / Techniques of aspiration cytology, biopsy and cell block of the bone marrow for the identification and determination of parasite intensity in Canine Visceral Leishmaniasis

O objetivo com este estudo foi comparar as técnicas de citologia aspirativa, biópsia e citobloco para identificação e quantificação parasitológica de Leishmania (Leishmania) infantum chagasi em medula óssea de cães. Amostras de tecido medular de 26 animais, em diferentes estágios clínico-laboratoriais da doença, foram estudadas obedecendo-se os mesmos critérios de investigação nas técnicas de citologia aspirativa, biópsia e citobloco. O menor número de campos para a confirmação parasitológica foi constatado no esfregaço direto obtido por citologia aspirativa. O estágio clínico-laboratorial não influenciou no número de campos necessários para a primeira visualização do agente em nenhuma das técnicas (p>0,05), e menor intensidade parasitária foi observada nas lâminas de citobloco. As técnicas de citologia aspirativa e biópsia concordaram na estimativa do coeficiente de infectividade no tecido estudado (p<0,05). Apesar de a técnica de citobloco permitir a concentração de células e o melhor reaproveitamento de amostras, não demonstrou ser um método adequado para rápida identificação e quantificação parasitológica na leishmaniose visceral canina. Considerando-se suas vantagens, a citologia aspirativa foi o melhor método para detecção microscópica do parasito e determinação do nível de intensidade parasitária no tecido estudado.(AU)

The aim of the present study was to compare the aspiration cytology, biopsy and cell block techniques for identification and parasitological quantification of Leishmania (Leishmania) infantum chagasi in dog bone marrow. Bone marrow tissue samples from 26 animals, in different clinical-laboratory stages of the disease, were studied according to the same criteria of investigation in the aspiration cytology, biopsy and cell block techniques. The lowest number of fields for the parasitological confirmation was found in the direct smear obtained by aspiration cytology. The clinical-laboratory stage did not influence the number of fields required for the first visualization of the agent in any of the techniques (p> 0.05) and less parasitic intensity was observed in the cell block slides. The aspiration cytology and biopsy techniques agreed on the estimation of infectivity coefficient in the tissue studied (p< 0.05). Although the cell block technique allows the concentration of cells and better reutilization of samples, it has not been shown to be an adequate method for rapid identification and parasitological quantification in canine visceral leishmaniasis. Considering its advantages, aspiration cytology was the best method for microscopic detection of the parasite and determination of the level of parasite intensity in the tissue studied.(AU)

Detection and molecular characterization of Trypanosoma (Duttonella) vivax in dairy cattle in the state of Sergipe, northeastern Brazil / Detecção e caracterização molecular de Trypanosoma (Duttonella) vivax em gado leiteiro no estado de Sergipe, no Nordeste do Brasil

Abstract Trypanosoma (Duttonella) vivax is an important cause of economic losses among feedlot cattle. These losses are related to the morbidity, mortality, reproductive issues and decreased production. It is known that the clinical signs observed in infections by this protozoon are similar to other hemoparasitosis, which difficult the diagnosis. Therefore, the aim of this study was to detect and molecularly characterize an outbreak of trypanosomiasis caused by T. (D.) vivax in dairy cattle in the municipality of São Miguel Aleixo, state of Sergipe, Brazil. Blood samples from cattle (n = 15) presenting clinical signs compatible with trypanosomiasis were collected and parasitological and molecular evaluated. Among the samples analyzed, 34% (5/15) were positive from blood smears, 60% (9/15) from the buffy coat method and 80% (12/15) from the molecular method. The DNA sequence obtained (659 bp) showed 99% similarity to T. (D.) vivax sequences that are available in the GenBank database. The presence of this protozoon in cattle herds is a problem for producers. Diagnosing trypanosomiasis is problematic because its evolution is similar to that of other parasitic blood diseases. In addition, this is the first report of infection by T. (D.) vivax in cattle in the state of Sergipe, northeastern Brazil.

Resumo Trypanosoma (Duttonella) vivax é responsável por consideráveis perdas econômicas na bovinocultura. Estas perdas estão relacionados à morbidade, mortalidade, problemas reprodutivos e declínio na produção. Sabe-se que os sinais clínicos apresentados em infecções por este protozoário se assemelha a outras hemoparasitoses, dificultando muitas vezes o diagnóstico. Portanto, objetivou-se com este estudo detectar a ocorrência de T. (D.) vivax em bovinos leiteiros no município de São Miguel Aleixo, Estado de Sergipe, Brasil. Para tanto, amostras de sangue (n = 15) foram coletadas e avaliadas através de métodos parasitológicos e moleculares. Do total das amostras analisadas, 34% (5/15) foram positivas no esfregaço sanguíneo, 60% (9/15) pelo método do Buffy Coat, enquanto na biologia molecular 80% (12/15) amplificaram um fragmento de DNA (659 pb) compatível com T. (D.) vivax (GenBank). Em conclusão a presença de T. (D.) vivax nos rebanhos bovinos caracteriza-se como um problema para os pecuaristas, como também para o diagnóstico, uma vez que essa tripanossomíase apresenta evolução semelhante a outras hemoparasitoses. Ademais, este é o primeiro relato de infecção por T. (D.) vivax em bovinos do estado de Sergipe, nordeste do Brasil.

Diagnosis and management of bovine babesiosis outbreaks in cattle in Punjab state.

AIM: The aim of the present study was to diagnose severe outbreaks of bovine babesiosis in Punjab state, in the year 2015 and to suggest control and preventive measures to animal owners. MATERIALS AND METHODS: Mortality of animals was recorded in two cattle herd comprising a total of 465 cattle in Sangrur (n=125) and Faridkot (n=340) districts. There was a history of purchase of animals at one farm. 23 blood samples were collected from diseased (n=15) and healthy animals (n=8) for hematological analysis, parasitological, and polymerase chain reaction (PCR)-based diagnosis. Ticks were also collected from animals for identification. RESULTS: Out of 465 cattle at risk, 28 were critically ill and 14 died of disease with morbidity, mortality, and case fatality rate of 6.02%, 3.01%, and 50.00%, respectively. Clinical signs and necropsy findings were suggestive of babesiosis. Ticks collected from both the outbreaks were identified as Rhipicephalus (Boophilus) microplus. Thin blood smears from infected animals (especially with clinical sign of hemoglobinuria) were found positive for Babesia bigemina organisms; however, molecular diagnosis (PCR) further confirmed the disease. Animals were successfully treated with diminazene aceturate, hematinics, and antipyretics. CONCLUSIONS: Two fatal outbreaks of babesiosis in cattle were diagnosed with application of conventional parasitological, hematological, and molecular diagnostic techniques. PCR was found to be far more sensitive in detecting the disease, especially in latent infections. Animal owners were advised to follow quarantine measures before mixing new animals in the herd and strategic acaricidal treatments for effective tick control.

Coproparasitological survey of intestinal parasites in the city of Londrina, Parana, Brazil: a retrospective analysis

Intestinal parasites are a major public health problem. It is important to inform and educate the public about these infections, especially where such data are scarce. This study aimed to determine the prevalence of intestinal parasites from the analysis of medical records of individuals of the city of Londrina. We analyzed 11,641 fecal reports from February 2009 to December 2012. Data were cataloged after the completion of parasitological testing of Hoffmann, Pons & Janer, Faust and Kato-Katz. From 11,641 reports, 19.1% were positive for intestinal parasites. Among those, 52.1% pertained to females and 47.9% to males, with predominance of positivity of 27.1% among children 0-10 years. For the regions studied, the northern region stood out with 35.4% of cases and prevalence of 6.8%. Among the pathogenic protozoa, reports of Giardia lamblia comprised 19.1% of positivity, while hookworms were the most frequent among helminths, comprising 7.8% of positive cases. It follows that poor conditions of basic sanitation contribute to the dissemination of these parasites. Early diagnosis is a determinant of successful treatment. Additionally, epidemiological data may be used to study the risk factors for transmission and may result in measures applicable to improving living conditions in the community

Performance analysis of software for identification of intestinal parasites / Análise de desempenho de software para identificação de parasitas intestinais

ABSTRACTIntroduction:Intestinal parasites are among the most frequent diagnoses worldwide. An accurate clinical diagnosis of human parasitic infections depends on laboratory confirmation for specific differentiation of the infectious agent.Objectives:To create technological solutions to help parasitological diagnosis, through construction and use of specific software.Material and method:From the images obtained from the sediment, the software compares the morphometry, area, perimeter and circularity, and uses the information on specific morphological and staining characteristics of parasites and allows the potential identification of parasites.RESULTS:Our results demonstrate satisfactory performance, from a total of 204 images analyzed, 81.86% had the parasite correctly identified by the computer system, and 18.13% could not be identified, due to the large amount of fecal debris in the sample evaluated.Discussion:Currently the techniques used in Parasitology area are predominantly manual, probably being affected by variables, such as attention and experience of the professional. Therefore, the use of computerization in this sector can improve the performance of parasitological analysis.Conclusions:This work contributes to the computerization of healthcare area, and benefits both health professionals and their patients, in addition to provide a more efficient, accurate and secure diagnosis.

RESUMOIntrodução:As parasitoses intestinais figuram entre os diagnósticos mais frequentes no mundo. Um diagnóstico clínico acurado das infecções parasitárias humanas depende da confirmação laboratorial para diferenciação específica do agente infeccioso.Objetivos:Criar formas tecnológicas para auxiliar no diagnóstico parasitológico por meio da construção e da utilização de um software específico.Material e método:A partir das imagens obtidas do sedimento, o software compara a morfometria, a área, o perímetro e a circularidade, além de utilizar informações sobre características específicas de morfologia e coloração dos parasitos e permitir a identificação dos possíveis parasitas.Resultados:Nossos resultados apontam desempenho satisfatório, sendo que do total de 204 imagens analisadas, 81,86% tiveram o parasita identificado corretamente pelo sistema computacional e 18,13% não puderam ser identificados, em função da grande quantidade de detritos fecais na amostra avaliada.Discussão:Atualmente, as técnicas realizadas no setor de Parasitologia são predominantemente manuais, sendo afetadas possivelmente por variáveis como atenção e experiência do profissional. Portanto, a utilização da informatização deste setor pode melhorar a performance das análises parasitológicas.Conclusão:O presente trabalho contribui para a informatização da área da saúde e beneficia tanto os profissionais da saúde como também seus clientes, além de proporcionar um diagnóstico mais eficiente, preciso e seguro.

Leishmaniasis: diagnostic issues in Europe.

Leishmaniasis is a parasitic disease with clinical presentations that vary from asymptomatic infection to cutaneous, mucocutaneous or visceral disease. Recent epidemiological studies have shown an increased prevalence in Europe largely caused by an increase in international travel, difficulty eradicating leishmanial infection in AIDS patients, and the use of immunosuppressive medications. Clinical diagnosis may be challenging, and parasitological diagnosis entails the use of invasive procedures which may be unrevealing in the immunosuppressed. A number of less invasive tests for the detection of anti-leishmanial antibodies or leishmanial antigen are available but their sensitivity and specificity may vary with the infective species and results have to be interpreted in light of the clinical presentation. The availability of polymerase chain reaction assays amplifying leishmanial genetic material has been a major step forward in improving the diagnosis of leishmanial disease and the response to treatment.

Transición parasitaria: experiencia en un hospital pediátrico de tercer nivel (1990-2010) / Parasitological transition: Experience in a third-level pediatric hospital (1990-2010)

ResumenINTRODUCCIÓN: Hace algunos años, el motivo principal para la solicitud del diagnóstico parasitológico era la sospecha de una etiología de base parasitaria. En la actualidad, el motivo de consulta y la solicitud del diagnóstico se encuentran asociados con una comorbilidad hemato-oncológica, autoinmune, inmunodeficiente o una condición postrasplante.MÉTODO: Se analizaron los resultados del laboratorio de parasitología clínica del Hospital Infantil de México Federico Gómez de enero de 1990 a diciembre del 2010, para obtener la frecuencia y distribución de parásitos y comensales por año y década.RESULTADOS: Se analizaron 211,600 muestras del periodo estudiado, con una media anual de 10,100 y desviación estándar de 3,500. El porcentaje de parásitos y comensales fue del 37.73%, y la frecuencia de positividad solo a parásitos fue del 21.4%. La diferencia de proporciones entre décadas resultó significativa, con una p < 0.0001.CONCLUSIONES: La tendencia en el comportamiento de las parasitosis intestinales durante el periodo estudiado mostró un cambio tanto en frecuencia como en género y especie de los agentes reportados. La generación de información de carácter epidemiológico es necesaria para conocer la tendencia de las parasitosis y de sus variaciones a través del tiempo.

AbstractBACKGROUND: In recent years, the main reason for parasitological diagnosis was suspicion of a parasitic etiology. Currently, diagnosis and consultation are associated with co-morbidity with a hemato-oncological, autoimmune, immunodeficiency or post-transplant condition.METHODS: Results collected from the Parasitology Laboratory of the Hospital Infantil de Mexico Federico Gómez from January 1990 to December 2010 were analyzed to determine the frequency and distribution of parasites and commensals per year and decade.RESULTS: There were 211,600 samples analyzed with a statistical media of 10,100 and a standard deviation of 3,500 samples. Percentage of observations for parasitic structures as well as commensals was 37.73% according to Pearson Chi square with a significant difference among percentages between decades (p < 0.0001).CONCLUSIONS: Behavioral tendency of intestinal parasitosis during the last two decades shows a change in frequency and genera from the agents that were reported at this pediatric health care level. The processing of epidemiologic information is important in order to determine the response of the parasitosis and its variations over time.

Evaluation of Kato-Katz and spontaneous sedimentation methods for the diagnosis of platynosomiasis in Neotropical primates / Avaliação dos métodos de sedimentação espontânea e Kato-Katz para o diagnóstico da platinossomose em primatas neotropicais

Platynosomiasis is a parasitic infection reported in non-human primates, including marmosets, and is frequently difficult to diagnose. In this study, the Kato-Katz method and the spontaneous sedimentation method were evaluated for their usefulness in identifying Platynosomum eggs in fecal samples from Callithrix penicillata that naturally harbor Platynosomum illiciens. Spontaneous sedimentation allowed the diagnosis of 41.7% (5/12) and 66.7% (8/12) of infected marmosets from one and three slides, respectively, prepared from the same fecal sample. The examination of a single Kato-Katz thick smear detected 83.3% (10/12) of infection cases. The analysis of feces on three different days increased the rate of diagnosis, since 75% (9/12) and 100% (12/12) of the primates with platynosomiasis were identified using serial spontaneous sedimentation (3 slides/day) and the Kato-Katz method, respectively. The mean number of Platynosomum eggs per gram of feces determined via the Kato-Katz method was 71.7 (8-240). The spontaneous sedimentation method when performed in series is acceptable for the diagnosis of platynosomiasis. However, the Kato-Katz method, which was here used for the first time to detect this infection, has a higher diagnostic sensitivity and the advantage that a quantitative analysis of the eggs released in the host feces is possible.

A platinossomose é uma infecção parasitária relatada em primatas não-humanos, inclusive saguis, cujo diagnóstico é frequentemente difícil. Neste estudo, os métodos de sedimentação espontânea e Kato-Katz foram avaliados quanto à sua utilidade na identificação de ovos de Platynosomum em amostras fecais de Callithrix penicillata naturalmente albergando Platynosomum illiciens. A sedimentação espontânea permitiu o diagnóstico de 41,7% (5/12) e 66,7% (8/12) dos saguis infectados a partir da análise de uma e três lâminas, respectivamente, preparadas de uma mesma amostra fecal. O exame de uma única lâmina de Kato-Katz detectou 83,3% (10/12) dos casos de infecção. A análise de fezes em três dias diferentes aumentou as taxas de diagnóstico, uma vez que 75% (9/12) e 100% (12/12) dos primatas que apresentaram a platinossomose foram identificados, usando-se a sedimentação espontânea (três lâminas/dia) e o Kato-Katz em série, respectivamente. O número médio de ovos de Platynosomum por g de fezes, determinado através do método de Kato-Katz, foi de 71,7 (8-240). O método de sedimentação espontânea, quando realizado em série, é aceitável para o diagnóstico da platinossomose. Entretanto, o método de Kato-Katz, o qual foi pela primeira vez usado para se detectar essa infecção, mostrou uma maior sensibilidade diagnóstica, com a vantagem de que é possível uma análise quantitativa dos ovos liberados nas fezes do hospedeiro.

Membrane fractions from Strongyloides venezuelensis in the immunodiagnosis of human strongyloidiasis / Frações de membrana de Strongyloides venezuelensis para o imunodiagnóstico da estrongiloidíase humana

Strongyloides venezuelensis is a parasitic nematode of rodents frequently used to obtain heterologous antigens for the immunological diagnosis of human strongyloidiasis. The aim of this study was to evaluate membrane fractions from S. venezuelensis for human strongyloidiasis immunodiagnosis. Soluble and membrane fractions were obtained in phosphate saline (SS and SM) and Tris-HCl (TS and TM) from filariform larvae of S. venezuelensis. Ninety-two serum samples (n = 92) were obtained from 20 strongyloidiasis patients (Group I), 32 from patients with other parasitic diseases (Group II), and 40 from healthy individuals (Group III), and were analyzed by enzyme-linked immunosorbent assay (ELISA). Soluble fractions (SS and TS) showed 90.0% sensitivity and 88.9% specificity, whereas the membrane fractions (SM and TM) showed 95.0% sensitivity and 94.4% specificity. The present results suggest the possible use of membrane fractions of S. venezuelensis as an alternative antigen for human strongyloidiasis immunodiagnosis.

Strongyloides venezuelensis é um nematódeo parasita de roedores, frequentemente usado como antígeno heterólogo para o diagnóstico imunológico da estrongiloidíase humana. O objetivo deste estudo foi avaliar frações de membrana de S. venezuelensis para o imunodiagnóstico da estrongiloidíase humana. Para tanto, frações solúveis e de membrana foram obtidas em solução salina fosfato (SS e MS) e Tris-HCl (ST e MT) de larvas filarioides de S. venezuelensis. Amostras de soro de 92 indivíduos, sendo 20 com estrongiloidíase (Grupo I); 32 com outras parasitoses (Grupo II), e 40 indivíduos saudáveis (Grupo III), foram analisadas pelo teste Imunoenzimático (ELISA). As frações solúveis (SS e ST) apresentaram 90,0% e 88,9%, enquanto que as frações de membrana (MS e MT) demonstraram 95,0% e 94,4%, de sensibilidade e especificidade, respectivamente. Os resultados obtidos permitem indicar as frações de membranas como antígeno alternativo para o diagnóstico da estrongiloidíase humana.

[Parasitological transition: Experience in a third-level pediatric hospital (1990-2010)]. / Transición parasitaria: experiencia en un hospital pediátrico de tercer nivel (1990-2010).

BACKGROUND: In recent years, the main reason for parasitological diagnosis was suspicion of a parasitic etiology. Currently, diagnosis and consultation are associated with co-morbidity with a hemato-oncological, autoimmune, immunodeficiency or post-transplant condition. METHODS: Results collected from the Parasitology Laboratory of the Hospital Infantil de Mexico Federico Gómez from January 1990 to December 2010 were analyzed to determine the frequency and distribution of parasites and commensals per year and decade. RESULTS: There were 211,600 samples analyzed with a statistical media of 10,100 and a standard deviation of 3,500 samples. Percentage of observations for parasitic structures as well as commensals was 37.73% according to Pearson Chi square with a significant difference among percentages between decades (p<0.0001). CONCLUSIONS: Behavioral tendency of intestinal parasitosis during the last two decades shows a change in frequency and genera from the agents that were reported at this pediatric health care level. The processing of epidemiologic information is important in order to determine the response of the parasitosis and its variations over time.

Balantidium coli and other gastrointestinal parasites in captives non-human primates of the Rio de Janeiro, Brazil.

BACKGROUND: Parasites are agents of diarrhea in captive non-human primates (NHP). To broaden information about those etiological agents in captive animals in Brazil, gastrointestinal parasites in captive NHP were researched in nurturing Centers of Rio de Janeiro State. METHODS: Fecal samples were collected from primates, of which 960 came from the Research Center (Cecal/Fiocruz) and 115 from the Primate Center (CPRJ/Inea). The study involved species of the New World (NW) primates and of the Old World (OW). RESULTS: The estimated prevalence was 56.7%, of which 91.3% presented protozoans and 7.4% presented helminths. Statistical difference between the nurturing centers occurred in the overall value of parasitosis and in the isolated frequency of Balantidium coli and Entamoeba sp., especially in the samples of OW primates living in Cecal. CONCLUSIONS: These results demonstrated the need for implements of sanitation programs in the sites for captive primates nurturing.

Adult female of Strongyloides stercoralis in respiratory secretions.

OBJECTIVE: To communicate the presence of adult females, rabditoid larvae and eggs of Strongyloides stercoralis (S. stercoralis) in the respiratory secretions obtained by tracheal aspirate from a HIV-negative patient who was suffering from polymyositis, and treated with corticoids and amethopterin and assisted by pneumonia. METHODS: The respiratory secretions submitted to the Parasitology Laboratory of the Muñiz Hospital were made more concentrated by centrifugation (1 500 r/min for 15 seconds). Wet mount microscopy was performed with the pellet. RESULTS: It revealed adult females, rabditoid larvae and eggs of S. stercoralis. Further parasitological studies performed after the start of the treatment with ivermectin on fresh fecal samples, gastric lavages and tracheal aspirates showed scanty mobile filariform and rabditoid larvae of the same parasite. CONCLUSIONS: The presence of adult female S. stercoralis which has never been observed before in the clinical samples submitted to our Laboratory for investigation can be considered as an indirect marker of the severe immunosupression of the patient.

Evaluación de tres técnicas coproparasitoscópicas para el diagnóstico de geohelmintos intestinales / Evaluation of three coproparasitoscopic techniques for the diagnosis of intestinal geohelminthiases

Introducción: las geohelmintiasis son un problema de salud pública; sin embargo, para su diagnóstico no se ha establecido una técnica de referencia (estándar de oro) sino que se han empleado diferentes pruebas coproparasitoscópicas con distintas tasas de sensibilidad. Objetivo: determinar la sensibilidad de la técnica coproparasitoscópica de Kato-Katz frente a la combinación del examen directo y la concentración de Ritchie para detectar geohelmintos y establecer el grado de infección. Metodología: se comparó la técnica de Kato-Katz con la combinación del coprológico directo y por concentración de Ritchie, estas dos últimas tomadas como estándar de oro, en 90 muestras de materia fecal provenientes de niños del barrio La Cruz, localizado en el Nororiente de Medellín. Resultados: la sensibilidad de la técnica de Kato-Katz para el diagnóstico de infecciones por geohelmintos fue similar a la obtenida con la combinación del coprológico directo y por concentración. Conclusión: la técnica de Kato-Katz da resultados confiables con alta sensibilidad para el diagnóstico de las geohelmintiasis intestinales más frecuentes en Colombia.

Introduction: Soil transmitted helminths (geohelminthiases) are a worldwide public health problem. However, no reference technique (gold standard) for their diagnosis has been established; contrariwise, several coproparasitoscopic tests, with variable sensibility rates, have been used. Objective: To compare the sensibility of the coproparasitoscopic Kato-Katz technique with that of the combination of direct coprologic examination and Ritchie's concentration, for the detection of geohelminths, and to determine the degree of infection. Methodology: The comparison was made in 90 stool specimens obtained from school children living in a low-income community in Medellín, Colombia. Results: Sensibility of the Kato-Katz technique was similar to the one obtained with the combination of direct microscopic examination and coprology by concentration. Conclusion: The Kato-Katz technique provides reliable results with high sensibility rate for the diagnosis of the intestinal geohelminths more frequent in Colombia.

Adult female of Strongyloides stercoralis in respiratory secretions

Objective: To communicate the presence of adult females, rabditoid larvae and eggs of Strongyloides stercoralis (S. stercoralis) in the respiratory secretions obtained by tracheal aspirate from a HIV-negative patient who was suffering from polymyositis, and treated with corticoids and amethopterin and assisted by pneumonia. Methods: The respiratory secretions submitted to the Parasitology Laboratory of the Mu?iz Hospital were made more concentrated by centrifugation (1 500 r/min for 15 seconds). Wet mount microscopy was performed with the pellet. Results: It revealed adult females, rabditoid larvae and eggs of S. stercoralis. Further parasitological studies performed after the start of the treatment with ivermectin on fresh fecal samples, gastric lavages and tracheal aspirates showed scanty mobile filariform and rabditoid larvae of the same parasite. Conclusions: The presence of adult female S. stercoralis which has never been observed before in the clinical samples submitted to our Laboratory for investigation can be considered as an indirect marker of the severe immunosupression of the patient.