Association of toll-like receptors in malaria susceptibility and immunopathogenesis: A meta-analysis.

Toll-like receptors (TLRs) play a key role in the induced immune response in malaria. Although the potential roles of TLRs have been described, it is necessary to elucidate which of these receptors may actually have an impact on the immunopathogenesis of the disease. This article performed a meta-analysis adhered to the PRISMA statement on TLRs studied in malaria by Plasmodium falciparum and Plasmodium vivax and its impact on susceptibility and pathogenesis during malaria. A search of the literature was undertaken in PubMed, LILACS and SciELO published until June 30th, 2020. The risk of bias was calculated using the Joanna Briggs Institute's Critical Review Checklist. Later, based on the inclusion and/or exclusion criteria, 17 out of 296 articles were harvested for this systematic review, the meta-analysis included studies incorporating 6,747 cases and 8,983 controls. The results showed that only TLR1, TLR9 and TLR4 receptors were associated with parasitemia, TLR2 and TLR6 were related with severity and none TLR was correlated with susceptibility. The data described here should be taken with caution, since the current evidence is limited and inconsistent. More studies are needed given that the results may change depending on the region and genetic background of the populations.

Molecular characterization of Cryptosporidium spp. in dogs and cats in the city of Rio de Janeiro, Brazil, reveals potentially zoonotic species and genotype.

Intestinal cryptosporidiosis is a diarrheal disease caused by protists of genus Cryptosporidium that infect a wide variety of hosts, primarily vertebrates. Due to the close contact between humans and their companion animals, especially dogs and cats, there is concern about the potential for zoonotic transmission of this enteric protozoan parasite by infected animals. This study aimed to perform a microscopic and molecular diagnosis of Cryptosporidium spp. in fecal samples from domiciled dogs and cats. One hundred and nineteen fecal samples were processed using sugar centrifugal flotation followed by molecular detection of Cryptosporidium spp. DNA using nested PCR. Subtyping of isolates positive for C. parvum was performed by sequence analysis of the 60 kDa glycoprotein gene (GP60). Cryptosporidium oocysts were detected in 7.8% (5/64) and 5.4% (3/55) of the fecal samples from dogs and cats, respectively. Cryptosporidium canis (n = 3) and C. parvum (n = 2) were the main species found in dogs, whereas C. felis (n = 3) was prevalent in cats. Subtype IIaA17G2R2 (potentially zoonotic) was identified in samples positive for C. parvum. Despite the low prevalence of Cryptosporidium observed in the domiciled dogs and cats, the presence of potentially zoonotic C. parvum in dogs evidences a public health concern. Further research is needed to better understand the epidemiology, source, and potential impacts of Cryptosporidium infection in cats and dogs.

Occurrence of anti-Toxoplasma gondii antibodies and risk factors associated with infection in equids slaughtered for human consumption in Brazil.

Toxoplasmosis is a worldwide zoonosis caused by Toxoplasma gondii. Ingestion of raw/undercooked meat is considering an important route of infection. Consumption of meat from equids is common in European and Asian countries and an increase in Brazil has been observed. The aim of this study was to evaluate occurrences of anti-T. gondii antibodies and risk factors relating to infection in equids slaughtered for human consumption in Minas Gerais, Brazil. Blood samples from 192 horses and 208 donkeys were collected in the exsanguination area during the slaughter. Serum samples were subjected to the indirect fluorescent antibody test (IFAT). Association analysis was performed using Pearson's chi-square test (χ2) or Fisher's exact test, to evaluate risk factors relating to the prevalence of seroreagents. Antibodies against T. gondii were found in 13.5% of the equids, with higher occurrence in horses (18.75%) than in donkeys (8.65%). Associations between seropositivity and the following variables were found (p ≤ 0.05): species, animal origin, purpose of rearing and source of water for animal consumption and contact with cats. Farms need to implement preventive measures to control T. gondii infection in these species and avert transmission of the parasite to the human population that will consume their meat.

Increasing the Efficiency of Spontaneous Sedimentation for the Diagnosis of Intestinal Protozoa.

The aim of this study was to evaluate the association between a longer sedimentation time, the reading of a larger number of slides, and the collection of multiple samples on the efficiency of the spontaneous sedimentation technique. Twenty-two patients with a previous parasitological exam positive for intestinal protozoa were recruited to collect new fecal samples (3 samples per patient) before the beginning of antiparasitic treatment. All collected fecal samples were used for spontaneous sedimentation and centrifuge-flotation techniques. Of these, all 22 patients were positive based on spontaneous sedimentation, and 59.1% (13/22) based on centrifuge flotation. The number of samples and the number of slides analyzed by spontaneous sedimentation influenced the number of positive cases. The modifications applied to the spontaneous sedimentation technique increased its performance in protozoa diagnosis.

Frequency of enteroparasitoses in preschool children attending daycare centers: a survey applying parasitological and immunological methods

The present study evaluated the frequency of intestinal parasitoses in children in public day care centers applying parasitological and immunological diagnostic methods. Fecal samples from 121 children from six public daycare centers were analyzed using parasitological techniques. Epidemiological data were obtained through a questionnaire, where parents and / or guardians were asked, for instance, whether the children had contact with soil, ate raw food, such as vegetables or raw or undercooked meat, normally walked around barefoot or had contact with animals. Fecal samples from 82 children were also tested for Giardia intestinalis and Cryptosporidium sp. coproantigen using the enzyme-linked immunosorbent assay (ELISA) which was also used for Entamoeba coproantigen detection only in samples that tested positive for the parasite by parasitological stool exam/optical microscopy. Intestinal parasite infection was noted in 23.1% (28/121) of the children. The most frequent parasite was Giardia intestinalis (13.2%), followed by Entamoeba coli (5.8%), Blastocystis spp. (1.7%), Endolimax nana (1.7%), Enterobius vermicularis (1.7%), Cystoisospora belli (0.8%), Entamoeba histolytica/E. dispar complex (0.8%), and Ascaris lumbricoides (0.8%). Positivity for parasite infection using parasitological stool exams was significantly associated with age groups, with a higher frequency in 4 to 6 year old children (p=0.03). No association or significant variations were noted in the prevalence of intestinal parasites in relation to the epidemiological variables studied. All samples were negative for Cryptosporidium sp. and Entamoeba histolytica detected by immunological testing, and 17.1% (14/82) children tested positive for Giardia intestinalis, although using parasitological exam/optical microscopy, only 14.6% (12/82) tested positive. The high incidence of intestinal parasites, especially protozoans, suggests probable interpersonal transmission among the children, environmental contamination, or even contaminated food/water intake. Thus, consolidation of preventive measures and efficient diagnostic resources as well as control of intestinal parasites and patient treatment are of utmost importance

Toxoplasma gondii molecular and immunological identification and risk factors associated with infection in chicken slaughtered at Triângulo Mineiro region, Minas Gerais, Brazil / Identificação molecular e imunológica de Toxoplasma gondii e fatores de risco associados com infecção em galinhas abatidas na região do Triângulo Mineiro, Minas Gerais, Brasil

This study aimed to determine the occurrence of anti-Toxoplasma gondii antibodies in the serum of slaughtered chickens in the region of Triângulo Mineiro, Minas Gerais, Brazil, to detect the parasite in tissues (heart and brain) of serologically positive chickens, based on molecular analysis, and to investigate risk variables associated with the infection. Sera from 417 chickens raised in extensive, semi-intensive, and intensive production systems were tested by an indirect immunofluorescent antibody test (IFAT) and indirect hemagglutination antibody test (IHA). Polymerase chain reaction (PCR) was performed to detect T. gondii DNA in brain and heart tissues. Antibody anti-T. gondii were found in 37.65% (157/417) of chickens by IFAT, and in 75.06% (313/417) by IHA. The Kappa index showed a weak concordance between the techniques (0.087). Association was observed between seropositivity and the variables, age (p < 0.0001), type of feeding (p < 0.0001) and collective raising with other animal's species (p < 0.0001). Association, based on IFAT, was not observed between seropositivity and the variables, sex (p = 0.0526), presence of cats (p > 0.9999), and presence of rats (p > 0.9999). Presence of parasite DNA was detected in brain samples from two chickens, which were raised in intensive and semi-intensive production systems. The results suggest the meat of these slaughtered animals may serve as a transmission source of this protozoan to humans.(AU)

O objetivo do presente estudo foi determinar a frequência de anticorpos anti-Toxoplasma gondii em soro de galinhas abatidas na região do Triângulo Mineiro, Minas Gerais, detectar molecularmente o parasito em tecidos (coração e cérebro) de algumas das aves sorologicamente positivas e averiguar variáveis de risco associadas à infecção. Foram testados soros de 417 galinhas, criadas nos sistemas extensivo, semi-intensivo e intensivo. Para a pesquisa de anticorpos anti-T. gondii foi utilizada a Reação de Imunofluorescência Indireta (RIFI) e Hemaglutinação Indireta (HAI). A Reação em Cadeia da Polimerase (PCR) foi utilizada para detectar o DNA de T. gondii em fragmentos de cérebro e coração. Anticorpos foram detectados no soro de 37,65% (157/417) das aves pela RIFI e em 75,06% (313/417) pela HAI. O índice Kappa mostrou uma fraca concordância entre as técnicas (0,087). Baseado na RIFI, foi verificada associação estatisticamente significativa (p < 0,0001) entre a soropositividade e as variáveis: idade, tipo de alimentação e criação em conjunto com outras espécies animais. Não foi observada associação estatística (p > 0,01) entre as variáveis: sexo, presença de gatos e presença de ratos. Pelo diagnóstico molecular DNA do parasito foi detectado em duas amostras de cérebro, de indivíduos diferentes criados em sistema intensivo e semi-intensivo. Os resultados indicam a possibilidade de a carne dessas aves poderem atuar como fonte de infecção deste protozoário para o homem.(AU)

Assessment of the diagnostic performance of four methods for the detection of Giardia duodenalis in fecal samples from human, canine and feline carriers.

Enteric parasitic diseases including giardiasis are of public health concern. Different methods are available for the diagnosis of this parasitic infection in fecal samples such as the identification of protozoan cysts and trophozoites by light microscopy, detection of specific antigens by ELISA, and amplification of DNA fragments by PCR. The present study aimed at assessing the performance of four laboratory tests for the detection of Giardia duodenalis in fecal specimens from three different host species with a previous diagnosis of giardiasis; canine, feline and human patients provided new stool samples to be retested for Giardia before initiating treatment with antiprotozoal drugs. For this purpose, triplicate fecal specimens from 54 humans, 24 dogs and 18 cats living in the city of Niterói, RJ, southeast Brazil, were analysed by light microscopy, ELISA, immunochromatography, and nested PCR. The centrifugal-flotation method detected Giardia cysts in 89.6% (86/96) of the fecal samples. The protozoan parasite was detected via immunochromatography in 87.5% (84/96) of these samples. Giardia was detected by ELISA in 69.8% (67/96) of the stool specimens from carriers with a previous diagnosis of Giardia infection. Giardia was detected by PCR in only 39.6% (38/96) of the fecal specimens. Based on these findings, we suggest that, among the four assays that were used in this study, the zinc sulphate flotation technique (Faust et al., 1939) is the best diagnostic assay in terms of sensitivity and specificity to detect G. duodenalis on serially collected samples from dogs, cats and humans.

Captive-bred neotropical birds diagnosed with Cryptosporidium Avian genotype III.

Currently, there are only three valid species of Cryptosporidium infecting avian hosts, namely, Cryptosporidium meleagridis, Cryptosporidium baileyi, Cryptosporidium galli and Cryptosporidium avium in addition to 12 genotypes of unknown species status. The objectives of this study were to microscopically diagnose the presence of Cryptosporidium in birds from a commercial aviary located in Rio de Janeiro, Brazil; genotypically characterize species and/or genotypes of genus Cryptosporidum; and conduct sequencing and phylogenetic analyses to compare the obtained DNA sequences with those deposited in GenBank. A total of 85 fecal samples were collected from wild captive-bred birds: 48 of family Psittacidae and 37 of family Ramphastidae. Initially, a search for the presence of Cryptosporidium sp. oocysts was conducted using the centrifugal-flotation in saturated sugar solution technique, after that, the collected samples were analyzed microscopically. Cryptosporidium infections were only detected in 24.32% of samples belonging to the family Ramphastidae. DNA was extracted from positive samples and molecular diagnostics was applied targeting the 18S rRNA gene, followed by sequencing and phylogenetic analysis. The Cryptosporidium Avian genotype III was diagnosed in this study more closely related to the gastric species. This is the first record of Cryptosporidium Avian genotype III in order Piciformes and family Ramphastidae, where three host species (Ramphastus toco, Ramphastus tucanus, and Pteroglossus bailloni) were positive for the etiologic agent. Based on the molecular data obtained, these wild birds raised in captivity do not represent a source of human cryptosporidiosis, considering that Cryptosporidium Avian genotype III does not constitute a zoonosis.

The influence of serial fecal sampling on the diagnosis of giardiasis in humans, dogs, and cats.

Giardia infection is a common clinical problem in humans and pets. The diagnosis of giardiasis is challenging as hosts intermittently excrete protozoan cysts in their feces. In the present study, we comparatively evaluated two methods of serial fecal sampling in humans, dogs, and cats from Rio de Janeiro, Brazil. The Faust et al. technique was used to examine fecal specimens collected in triplicate from 133 patients (52 humans, 60 dogs, and 21 cats). Specimens from 74 patients were received from the group assigned to carry out sampling on consecutive days - 34 humans, 35 dogs, and 5 cats, and specimens from 59 patients were received from the group assigned to carry out sampling on non-consecutive, separate days - 18 human beings, 25 dogs, and 16 cats. G. duodenalis cysts were found in stools of 30 individuals. Multiple stool sampling resulted in an increase in the number of samples that were positive for Giardia in both groups. The authors therefore conclude that multiple stool sampling increases the sensitivity of the Faust et al . technique to detect G. duodenalis cysts in samples from humans, cats and dogs.

Knowledge about parasite infections among pregnant and postpartum women who attended a university hospital in Niteroi, Rio de Janeiro, Brazil

Parasite infections are frequent in Brazil during pregnancy. The aim of the present study was to assess the knowledge about parasite infections among pregnant and post-partum women, attending a public University Hospital in Niteroi, RJ, Brazil. Knowledge was assessed using a standardized interview and questionnaire. A total of 100 interviews were conducted (60 pregnant and 40 post-partum women). Educational achievement did not influence knowledge about parasites. The majority of the women interviewed stated that they habitually always washed hands after using the bathroom and before eating. Consumption of fruits and vegetables was prevalent, but 17% did not wash them before eating and 39% ate raw or undercooked meat. Educational achievement and number of pregnancies did not influence hygiene habits. Neither the late pre-natal care nor the number of pregnancies influenced knowledge about parasites. Also, there was no correlation between the initial date of pre-natal care and the number of correct answers in the questionnaire. Similarly, there was no association between the initial date of pre-natal care and receiving information about parasite infections. 79% of the women interviewed stated that they had never received any information about parasite infection and pregnancy, and the harm it could cause to them and/or their baby. Despite advances in pre-natal care assistance in the last decade, the lack of knowledge among pregnant and post-partum women regarding parasites and risk factors for infection still remains.

Programs for controlling congenital toxoplasmosis: study of current status in a brazilian municipality

Measures for congenital toxoplasmosis prevention may be performed at three levels. Primary prevention is characterized by education and public health programs, in which susceptible pregnant women receive guidance on sources of infection and on adopting preventive measures. The secondary level consists of serological screening during prenatal care. Finally, the tertiary level operates in already infected newborns, seeking to prevent clinical damage and late complications. This study aims to describe the situation of control measures for congenital toxoplasmosis in Niterói-RJ according to the National Health Promotion Policy. In Niterói, only secondary prevention is being implemented and there is no guidance on preventive measures to seronegative pregnant women. There is also no notification of congenital and acquired toxoplasmosis cases during pregnancy

Toxoplasmosis-related knowledge among pregnant and postpartum women attended in public health units in Niterói, Rio De Janeiro, Brazil.

The present study conducted a toxoplasmosis-related knowledge level survey with 400 pregnant and puerperal women attended in public health units in the municipality of Niterói, Rio de Janeiro. Only 111 (27.8%) women claimed to know about the disease. Most of them (n = 289; 72.2%) had never heard about toxoplasmosis nor knew how to prevent the infection by Toxoplasma gondii. A significant difference (p = 0.013) regarding the presence of anti-T. gondii IgG was observed between women who claimed to know about the disease and those who had never heard about it. These results highlight the importance of a systematic serological screening process for toxoplasmosis, as well as the importance of primary prevention by accurate information during prenatal care, an important Public Health action to be implemented.

Toxoplasmosis-rllated knowledge among pregnant nd postpartum women attended inn public health units in niterói, rio de janeiro, Braill / Conhecimento sobre toxoplasmose entre gestantes e puérperas atendidas na rede pública de saúde do município de Niterói, Rio de Janeiro, Brasil

The present study conducted a toxoplasmosis-related knowledge level survey with 400 pregnant and puerperal women attended in public health units in the municipality of Niterói, Rio de Janeiro. Only 111 (27.8%) women claimed to know about the disease. Most of them (n = 289; 72.2%) had never heard about toxoplasmosis nor knew how to prevent the infection by Toxoplasma gondii. A significant difference (p = 0.013) regarding the presence of anti-T. gondii IgG was observed between women who claimed to know about the disease and those who had never heard about it. These results highlight the importance of a systematic serological screening process for toxoplasmosis, as well as the importance of primary prevention by accurate information during prenatal care, an important Public Health action to be implemented.

Este estudo teve por objetivo realizar um levantamento do conhecimento de 400 gestantes e puérperas atendidas na rede pública de saúde do município de Niterói, Estado do Rio de Janeiro sobre a toxoplasmose. Apenas 111 (27,8%) participantes relataram conhecer a doença. A maioria 289 (72,2%) nunca ouviu falar sobre a toxoplasmose, principalmente sobre as formas de prevenção da infecção pelo Toxoplasma gondii. Foi encontrada diferença significativa entre as pacientes que disseram conhecer a toxoplasmose e aquelas que relataram desconhecer a doença com relação à presença de anticorpos IgG anti- T.gondii (p = 0,013). Estes resultados permitem ressaltar a importância do rastreamento sorológico sistemático na infecção toxoplásmica, bem como a prevenção primária por meio de informações corretas durante o pré-natal, sendo esta uma importante medida de saúde pública a ser implementada.

Seroepidemiology Toxoplasma gondii infection in students of veterinary medicine and other courses of public universities in Rio de Janeiro state, Brazil

The aim of this study was to estimate the serum prevalence of Toxoplasma infection in two groups ofuniversity students: Veterinary Medicine (VM) and Other Courses (OC). Students from two public universities in the state of Rio de Janeiro were investigated to identify risk habits and behaviors for Toxoplasma gondii infection, and to guide primary prevention. All 839 students answered aquestionnaire about habits and behaviors (347 OC and 492 VM). A serum prevalence of 21.8 percent was observed by indirect immunofluorescence (IFA) and ELISA. No statistically significant difference was observed between students of the universities. Serum prevalence (IFA and ELISA IgG) was16.1 percent in VM and 29.9 percent in OC. Six students were IgM seropositive, of which five were onlypositive in ELISA and one was positive with both diagnostic techniques (ELISA and IFA). Amongthose students, four were in the control group from UFF and one in each group from UFRRJ. The prevalence of students seropositive for toxoplasmosis was low in both universities and the resultsobtained suggest that veterinary students are probably not exposed to a higher risk than the generalpopulation for acquiring toxoplasmosis. Age, contact with cats, consumption of undercooked orraw meat, contact with campus soil and ignorance of prophylactic measures for toxoplasmosis werepositively associated with prevalence of infection by T. gondii.

O objetivo deste estudo foi estimar a soroprevalência da infecção toxoplásmica em dois grupos de universitários, de Medicina Veterinária (MV) e de Outros Cursos (OC), de duas instituições públicas de ensino do Estado do Rio de Janeiro, buscando identificar hábitos e comportamentos de risco para infecção por este protozoário, e orientando sobre a prevenção primária. Dos 839 universitários, 492 eram da MV e 347 de OC. Todos os acadêmicos responderam a um questionário sobre seus hábitos e costumes. A soroprevalência (RIFI e ELISA IgG) foi de 16,1 por cento na MV e 29,9 por cento nos OC. Seis estudantes foram IgM soro reagentes, dos quais cinco eram apenas positivos no ELISA e um era positivo em ambas as técnicas de diagnóstico (ELISA e RIFI). Entre os estudantes, quatro eram do grupo de controle da UFF e um em cada grupo da UFRRJ. A prevalência de universitários soros reagentes para a toxoplasmose nas duas Universidades foi baixa e os resultados obtidos neste estudo sugerem que provavelmente os estudantes de Medicina Veterinária não estão expostos a um risco maior que os de outros cursos de adquirir a toxoplasmose. A idade, o contato com gatos, com o solo do campus, o consumo de carne crua ou mal-passada e o desconhecimento sobre a profilaxia da toxoplasmose influenciaram na prevalência da infecção por T. gondii.

Estudo da contaminação de moedas e cédulas de dinheiro circulantes na cidade de Niterói-RJ / Survey of Parasite contamination of in Coins and Bills Circulating in the City of Niterói, Rio de Janeiro State

Estudos epidemiológicos demonstram elevada prevalência de parasitos intestinais no Brasil. Objetos inanimados, especialmente aqueles com grande circulação, são importantes veiculadores de formas de transmissão de parasitos capazes de resistir no meio ambiente. Portanto, o objetivo desteestudo foi verificar a frequência de contaminação parasitária em moedas e cédulas coletadas em estabelecimentos comerciais de grande circulação monetária na cidade de Niterói-RJ. A coleta e o processamento das amostras foram realizados segundo Levai et al. (1986), com pequenas alterações.Um total de 89 amostras (60 cédulas e 29 grupos de moedas) foi coletado em estabelecimentos comerciais de diferentes bairros de Niterói-RJ. Cédulas de 2,00 reais e 5,00 reais foram coletadas e armazenadas separadamente em sacolas plásticas até seu processamento. As moedas de 0,50 centavos, 0,10 centavos e 1,00 real foram coletadas de cada estabelecimento, armazenadas e processadas em conjunto. Foram analisadas cinco lâminas de cada amostra por microscopia óptica. Das 60 cédulas analisadas, 4 (6,7por cento) foram positivas para larvas de nematóides e 4 (6,7por cento) para artrópodes. As larvas observadasnestas amostras estavam parcialmente alteradas, não permitindo uma melhor identificação. Uma nota de 5,00 reais estava contaminada por artrópode e por ovo de nematelminto, o qual continha uma larva parcialmente alterada em seu interior. Todas as moedas analisadas foram negativas. Entre as amostras contaminadas, 62,5por cento foram coletadas em estabelecimentos que comercializavam alimentos. A presença de larvas de nematoides, artrópodes e de um ovo de nematelminto larvadoindica que as cédulas e moedas, devido a sua circulação rápida e indiscriminada, podem ser potenciais carreadores de estruturas parasitárias capazes de infectar seres humanos.

Ectopic dirofilariosis in two dogs from Rio de Janeiro state, Brazil / Dirofilariose ectópica em dois cães do estado do Rio de Janeiro, Brasil

Report of two canine dirofilariosis cases of ectopic location in the state of Rio de Janeiro. This is the first report of erratic migration for this parasitosis in dogs in the state, calling attention to the short period of time between the two cases. The fact that the area is endemic for this parasite, its zoonotic potential and the report of human cases in the state, demonstrates that authorities should be alerted to the control programs of dirofilariosis along with the pathogenic profile of the infections.

Relato de dois casos de dirofilariose canina de localização ectópica no Estado do Rio de Janeiro. Este é o primeiro relato de migração errática para esta parasitose em cães no Estado, e chama a atenção o curto espaço de tempo entre os dois casos. O fato da região ser endêmica para a parasitose, seu caráter zoonótico e o relato de casos humanos no Estado, demonstram que as autoridades competentes devem estar atentas aos programas de controle da dirofilariose assim como no perfil patogênico da infecção.

Ectopic dirofilariosis in two dogs from Rio de Janeiro state, Brazil.

Report of two canine dirofilariosis cases of ectopic location in the state of Rio de Janeiro. This is the first report of erratic migration for this parasitosis in dogs in the state, calling attention to the short period of time between the two cases. The fact that the area is endemic for this parasite, its zoonotic potential and the report of human cases in the state, demonstrates that authorities should be alerted to the control programs of dirofilariosis along with the pathogenic profile of the infections.