Dogs naturally infected by Rangelia vitalii, Babesia canis vogeli, and Ehrlichia canis in São Paulo, Brazil / Cães naturalmente infectados por Rangelia vitalii, Babesia canis vogeli, e Ehrlichia canis em São Paulo, Brasil

Several agents can cause hemoparasitic diseases in dogs, and blood-sucking arthropods transmit these diseases. These agents can cause several clinical manifestations and, in some cases, can kill the host. Because these agents are essential in animal health, this study aims to detect the frequency of Ehrlichia canis, Rickettsia rickettsii, Anaplasma platys, and Rangelia vitalii by real-time PCR and Babesia vogeli in dogs in the southern region of the city of São Paulo, São Paulo. Of the 98 dog samples, 18 (18.4%) tested positive with real-time polymerase chain reaction for at least one studied agent. Of these 18 samples, 17 tested positive for a single agent (11.2% for B. canis vogeli, 1.02% for R. vitalii, and 5.1% for E. canis), and one showed co-infection with B. canis vogeli and R. vitalii. The results demonstrate the presence of hemoparasites in the studied animals, which can influence the quality and life expectancy of these animals. The Rangeliadetection warns small animal clinicians to include it as a differential diagnosis for hemoparasitosis.(AU)

As hemoparasitoses em cães podem ser causadas por diversos agentes, sendo essas doenças transmitidas por artrópodes hematófagos. Esses agentes podem causar diversas manifestações clínicas e, em alguns casos, podem matar o hospedeiro. Este estudo teve como objetivo detectar por PCR em tempo real a frequência de Ehrlichia canis, Rickettsia rickettsii, Anaplasma platys, Rangelia vitalii e Babesia canis vogeli em amostras de cães da zona sul da cidade de São Paulo, Brasil. Das 98 amostras de cães, 18 (18,4%) testaram positivo com reação em cadeia da polimerase em tempo real para pelo menos um agente estudado. Destas 18 amostras, 17 testaram positivo para um único agente (11,2% para B. canis vogeli, 1,02% para R. vitalii e 5,1% para E. canis), e uma apresentou coinfecção com B. canis vogeli e R. vitalii. Os resultados demonstram a presença de hemoparasitas nos animais estudados, o que pode influenciar a qualidade e a expectativa de vida desses animais. Além disso, é o primeiro relato da detecção de R. vitalli na zona sul de São Paulo e serve de alerta para os clínicos de pequenos animais incluírem esse agente como diagnóstico diferencial para as hemoparasitoses.(AU)

Prevalence and Risk Factors for Intestinal Protozoan Infections in Children with Cancer in Taif, Western Saudi Arabia.

BACKGROUND: Cancer is a category of diseases that cause an individual's immune system to become suppressed. In a case-control study, the current study aims to detect the frequency of intestinal parasites and related risk factors in children with cancer. METHODS: Stool samples were collected from 178 children with cancers (cases) and 150 cancer-free children (controls) who sought treatment for diarrheal episodes at nearby hospitals. Samples were processed by direct smear examination, concentration technique, permanent staining by Lugol's iodine, modified Ziehl-Neelsen, modified trichrome, and chromotrope 2R stains. RESULTS: The overall prevalence of intestinal parasites was 7.3% (24/328), with non-statistically significant differences between cases (7.8%; 14/178) and controls (6.6%; 10/150). Children with leukemia had a higher infection rate (9%; 9/100) than children with lymphoma (6.9%; 3/43) or solid tumors (5.7%; 2/35). Blastocystis sp. (3.3%) was the most common intestinal parasite found in cases, followed by Cryptosporidium sp. (2.2%), Giardia lamblia (1.6%), and Microsporidia sp. (0.5%). For all parasites, no statistical difference was found between the two groups. (p > 0.05). Male gender, young age, non-bottled water use, travel to parasite-endemic areas, living in an urban area, and infrequent hand washing were all associated with intestinal parasitosis, with non-statistical significance observed between the two groups. In children with cancer, intestinal parasites were found to be significantly associated with chronic (p = 0.04) and severe (p = 0.03) diarrhea. CONCLUSIONS: Children with cancer, particularly those with hematological cancers, should be screened for intestinal parasites on a regular basis and treated for their overall health.

Molecular detection of Apicomplexa protozoa in tissues from Alouatta guariba clamitans / Detecção molecular de protozoários Apicomplexa em tecidos de Alouatta guariba clamitans

The brown howler monkey (Alouatta guariba clamitans) is a primate species widely distributed in South America. Infections by protozoa are common in primates. However, studies on protozoa in primates in Brazil are scarce, so the goal of this study was to investigate DNA from the apicomplexan protozoa Neospora caninum, Sarcocystis spp. and Toxoplasma gondii in tissues of A. guariba clamitans. DNA extraction was performed on tissue samples from the heart, brain, liver, spleen, lung and intestine of six A. guariba clamitans from Santa Maria, Central Region of Rio Grande do Sul, Brazil. Conventional PCR was performed using 18S rRNA gene general primers for Apicomplexa and also specific primers to amplify Neosporaspp. and Toxoplasma gondii DNA. All animals were positive in the 18S PCR and the genetic sequencing confirmed the presence of Sarcocystis spp. DNA in the tissues of four animals belonging to at least two species (S. neurona and S. gigantea) and T. gondii DNA in the other two animals. One positive sample for T. gondii was genotypically characterized as atypical by the restriction fragment length polymorphism technique. N. caninum DNA was not detected in the tested samples. The presence of Apicomplexa protozoan DNA in the tissues of the six animals tested in this study highlights the importance of howler monkeys as maintainers of these pathogens in nature.(AU)

O bugio ruivo (Alouatta guariba clamitans) é uma espécie de primata amplamente distribuída na América do Sul. As infecções por protozoários são comuns em primatas. Entretanto, estudos sobre protozoários em primatas no Brasil são escassos, portanto o objetivo deste estudo foi pesquisar DNA dos protozoários Apicomplexa Neospora caninum, Sarcocystisspp. e Toxoplasma gondii em tecidos de A. guariba clamitans. A extração de DNA foi realizada em amostras de tecido do coração, cérebro, fígado, baço, pulmão e intestino de seis A. guariba clamitans oriundos de Santa Maria, Região Central do Rio Grande do Sul, Brasil. Foi realizada PCR convencional utilizando primers geral do gene 18S rRNA para Apicomplexa e também primers específicos para amplificação de DNA de Neospora spp.e Toxoplasma gondii. Todos os animais foram positivos no PCR geral para Apicomplexa e no sequenciamento genético confirmou-se a presença de DNA de Sarcocystis nos tecidos de quatro animais pertencentes a pelo menos duas espécies (S. neurona e S. gigantea), e DNA de T. gondii foi detectado nos outros dois animais. Uma amostra positiva para T. gondii foi caracterizada genotipicamente como atípico pela técnica de polimorfismo do comprimento do fragmento de restrição. Não foi detectado DNA de N. caninum nas amostras testadas. A presença de DNA de protozoários apicomplexa nos tecidos dos seis animais testados neste estudo destaca a importância dos bugios ruivos como mantenedores desses patógenos na natureza.(AU)

Urbanorum spp novo parasita no Brasil / Urbanorum spp new parasite in Brazil / Urbanorum spp nuevo parásito en Brasil

Introdução: As enteroparasitoses são foco de investigações científicas no mundo todo. Urbanorumspp. foi reconhecido como parasita em 1994 no Peru, expandindo-se pela América do Sul. Relatado pela primeira vez no Brasil em 2018, Maranhão. Este relato apresenta o segundo caso no estado do Paraná. Relato de caso: Paciente masculino, 56 anos, 75kg, diabético, habitante de São José dos Pinhais, área urbana. Procura atenção primária por dor ao evacuar, tenesmo e cólica abdominal. Nega diarréia, febre, sangue nas fezes e viagem recente. Exame físico abdominal, hemograma e parcial de urina sem alterações. Parasitológico de fezes: Urbanorum spp. Prescrito Nitazoxanida 500mg 12/12h por 3 dias. Paciente retorna com melhora da sintomatologia e parasitológico de controle negativo. Conclusão: Atualmente a escassez de estudos primários prospectivos dificultam o delineamento clínico-epidemiológico e tratamento da parasitose. A disseminação do parasita entre extremos do país em curto intervalo de tempo, aliada à carência de saneamento básico criam um alerta para seu grande potencial epidêmico. Por isso, as políticas de saúde pública devem priorizar ações informativas e preventivas a fim de evitar surtos e complicações. A atenção primária à saúde é fundamental nesse contexto, justamente pela longitudinalidade e abrangência do cuidado.

Background: Enteroparasitosis are the focus of scientific research worldwide. Urbanorum spp. was recognized as a parasite in Peru in 1994, expanding throughout South America. Reported for the first time in Brazil, state of Maranhão, in 2018. This report presents the second case in the state of Paraná. Case report: Male patient, 56 years old, 75kg, diabetic, inhabitant of São José dos Pinhais, urban area, seeks primary care for pain on bowel movement, tenesmus and abdominal cramps. Denies diarrhea, fever, bloody stools, recent trip. Abdominal examination, blood count and partial urine without changes. Stool parasitology: urbanorum spp. Prescribed Nitazoxanide 500mg 12/12h for 3 days. Patient returns with improvement of symptomatology and parasitological negative control. Conclusion:Currently, the scarcity of prospective studies and meta-analyzes make clinical-epidemiological design and treatment of parasitosis difficult. The spread of the parasite between extremes of the country in a short period of time, coupled with the lack of basic sanitation create a warning for its great epidemic potential. Therefore, public health policies should prioritize informative and preventive actions in order to avoid outbreaks and complications. Primary health care is fundamental in this context, precisely because of the longitudinally and comprehensiveness of care.

Introducción: Las enteroparasitosis el punto de enfoque de investigaciones científicas en todo el mundo. Urbanorum spp fue reconocido cómo parásito en 1994 en el Peru, expandiéndose en América do Sul. Relatado por primera vez en Brasil, Maranhão, 2018. Este informe se encuentra en segundo lugar en el estado de Paraná. Relato del caso: Paciente masculino, 56 años, 75 kg, diabético, habitante de São José dos Pinhais, área urbana. Búsqueda atención primaria por dolor al defecar, tenesmo, y dolor abdominal. Nega diarrea, fiebre, sangre en heces o viaje reciente. Examen físico abdominal, hemograma e tests de orina sin modificaciones. Análisis parasitología: urbanorum spp. Prescripto Nitazoxanide 500mg 12/12h durante 3 días. Paciente volvió con alivio sintomático e materia fecal negativo. Conclusión: En la actualidad la escasez de estudios prospectivos y metanálisis dificultan la delineación clínico-epidemiológica y el tratamiento de la parasitosis. La diseminación del parásito entre los extremos del país en un corto período de tiempo, junto con la falta de saneamiento básico, crea una alerta por su gran potencial epidémico. Por lo tanto, las políticas de salud pública deben priorizar las acciones informativas y preventivas para evitar brotes y complicaciones. La atención primaria de salud es fundamental en este contexto, precisamente por la longitudinalidad y la amplitud de la atención.

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.

[Lung infection by lophomonas spp. in a female patient with acute myeloid leukemia]. / Infección pulmonar por Lophomonas sp en una paciente con leucemia mieloide aguda.

Protozoic infections are prevalent worldwide, particularly in immunosuppressed patients. We reported the case of a patient from the city of Viña del Mar, Chile, a carrier of acute myeloid leukemia in whom an infection by Lophomonas sp. was confirmed by bronchoalveolar lavage. She was treated with antibiotics but died of complications of the underlying disease. There is little literature available on this microorganism. We conclude that Lophomonas sp. should be considered as a diagnostic possibility if protozoa are found in bronchoalveolar lavage of immunosuppressed patients.

Las infecciones por protozoos son prevalentes a nivel mundial, en particular en pacientes inmunosuprimidos. Comunicamos el caso de una paciente procedente de la ciudad de Viña del Mar, Chile, portadora de leucemia mieloide aguda en quiense confirmó una infección por Lophomonas sp. en lavado bronquioalveolar. Se manejó con antibióticos, pero falleció decomplicaciones de su enfermedad de base. Existe poca literatura disponible respecto a este microorganismo. Concluimos que debe considerarse a Lophomonas sp. como posibilidad diagnóstica si se encuentran protozoos en lavados bronquioalveolares de pacientes inmunosuprimidos.

Infección pulmonar por Lophomonas sp en una paciente con leucemia mieloide aguda / Lung infection by lophomonas spp. in a female patient with acute myeloid leukemia

RESUMEN Las infecciones por protozoos son prevalentes a nivel mundial, en particular en pacientes inmunosuprimidos. Comunicamos el caso de una paciente procedente de la ciudad de Viña del Mar, Chile, portadora de leucemia mieloide aguda en quiense confirmó una infección por Lophomonas sp. en lavado bronquioalveolar. Se manejó con antibióticos, pero falleció decomplicaciones de su enfermedad de base. Existe poca literatura disponible respecto a este microorganismo. Concluimos que debe considerarse a Lophomonas sp. como posibilidad diagnóstica si se encuentran protozoos en lavados bronquioalveolares de pacientes inmunosuprimidos.

ABSTRACT Protozoic infections are prevalent worldwide, particularly in immunosuppressed patients. We reported the case of a patient from the city of Viña del Mar, Chile, a carrier of acute myeloid leukemia in whom an infection by Lophomonas sp. was confirmed by bronchoalveolar lavage. She was treated with antibiotics but died of complications of the underlying disease. There is little literature available on this microorganism. We conclude that Lophomonas sp. should be considered as a diagnostic possibility if protozoa are found in bronchoalveolar lavage of immunosuppressed patients.

Development of nanoparticle-assisted PCR assay in the rapid detection of brain-eating amoebae.

Brain-eating amoebae (Acanthamoeba spp., Balamuthia mandrillaris, Naegleria fowleri) have gained increasing attention owing to their capacity to produce severe human and animal infections involving the brain. Early detection is a pre-requisite in successful prognosis. Here, we developed a nanoPCR assay for the rapid detection of brain-eating amoebae using various nanoparticles. Graphene oxide, copper and alumina nanoparticles used in this study were characterized using Raman spectroscopy measurements through excitation with a He-Ne laser, while powder X-ray diffraction patterns were taken on a PANanalytical, X'Pert HighScore diffractometer and the morphology of the materials was confirmed using high-resolution transmission electron microscopy (HRTEM). Using nanoparticle-assisted PCR, the results revealed that graphene oxide, copper oxide and alumina nanoparticles significantly enhanced PCR efficiency in the detection of pathogenic free-living amoebae using genus-specific probes. The optimal concentration of graphene oxide, copper oxide and alumina nanoparticles for Acanthamoeba spp. was determined at 0.4, 0.04 and 0.4 µg per mL respectively. For B. mandrillaris, the optimal concentration was determined at 0.4 µg per mL for graphene oxide, copper oxide and alumina nanoparticles, and for Naegleria, the optimal concentration was 0.04, 4.0 and 0.04 µg per mL respectively. Moreover, combinations of these nanoparticles proved to further enhance PCR efficiency. The addition of metal oxide nanoparticles leads to excellent surface effect, while thermal conductivity property of the nanoparticles enhances PCR productivity. These findings suggest that nanoPCR assay has tremendous potential in the clinical diagnosis of parasitic infections as well as for studying epidemiology and pathology and environmental monitoring of other microbes.

Evaluation of two DNA extraction methods for the PCR-based detection of eukaryotic enteric pathogens in fecal samples.

OBJECTIVE: Efficient and easy-to-use DNA extraction and purification methods are critical in implementing PCR-based diagnosis of pathogens. In order to optimize the routine clinical laboratory diagnosis of eukaryotic enteric pathogens, we compare, via quantitative PCR cycle threshold (Ct) values, the efficiency of two DNA extraction kits: the semi-automated EZ1® (Qiagen) and the manual QIAamp® DNA Stool Mini Kit (Qiagen), on six protozoa: Blastocystis spp., Cryptosporidium parvum/hominis, Cyclospora cayetanensis, Dientamoeba fragilis, Giardia intestinalis and Cystoisospora belli and one microsporidia: Enterocytozoon bieneusi. RESULTS: Whereas EZ1® (Qiagen) and QIAamp® DNA Stool Mini Kit (Qiagen) yielded similar performances for the detection of Cryptosporidium spp. and D. fragilis, significant lower Ct values (p < 0.002) pointed out a better performance of EZ1® on the five remaining pathogens. DNA extraction using the semi-automated EZ1® procedure was faster and as efficient as the manual procedure in the seven eukaryotic enteric pathogens tested. This procedure is suitable for DNA extraction from stools in both clinical laboratory diagnosis and epidemiological study settings.

EPIDEMIOLOGY OF CLOSTRIDIUM DIFFICILE INFECTION IN HOSPITALISED PEDIATRIC PATIENTS IN GEORGIA.

Clostridium difficile, a Gram-positive spore-forming bacillus, is the most common identifiable etiologic agent of antibiotic-associated diarrhea. The incidence of Clostridium difficile infections among hospitalized children has been increasing across the world. The aim of our study was to evaluate occurrence of Clostridium difficile and some other gastrointestinal pathogens among hospitalized pediatric patients in Georgia, as far as currently statistical data on the topic is very limited in the country. One of the objectives of the study was to test and pilot the real-time Polymerase Chain Reaction diagnostic systems for rapid and simultaneous identification of number of pathogens with a particular emphasis on diarrheal disease diagnostics as these are one of the primary public health priorities in Georgia and worldwide. Cross-Sectional study has been performed on 211 samples collected from 192 pediatric patients. Two pediatric hospitals were involved in the study: M. Iashvili Children's Central Hospital and Tbilisi Children's Clinical Hospital for Infectious Diseases. Laboratory investigations were done in the Clinic NeoLab, Tbilisi, Georgia. Study materials collected for testing were stool samples. Samples were tested by EIA kits (CerTest biotec, Zaragoza, Spain) for presence of A/B toxin according to the manufacturer's instructions. EIA test positive samples were analyzed by home-made multiplex real-time polymerase chain reaction (NeoPCR Diagnostics, NeoLab, Tbilisi, Gerogia) for confirmation of the infection and for simultaneous identification of additional gastrointestinal pathogens including Entamoeba histolitica, Giardia lamblia, Cryptsporidium parvum, Adenovirus, Rotavirus, Norovirus and Astrovirus. All samples were also tested for the presence of the above listed pathogenic agents using the same type EIA kits as for Clostridium difficile described above (CerTestbiotec, Zaragoza, Spain) for presence of the corresponding pathogen. The average age of the study participants was 3.5 years, 56.7% were male and 43.3% were female patients. Presence of Clostridium difficile have been documented in 21 samples out of 211 (10%). Besides the Clostridium difficile, other enteric pathogens have been revealed with the following frequencies: Rotavirus in 12 cases (5.7%), Adenovirus in 11 (5.2%), Giardia lamblia in 10 (4.7%), Astrovirus in 3 (1.4%), Cryptsporidium parvum in 3 (1.4%), Entamoeba histolitica in 2 (0.9%), Norovirus in 2 (0.9%). 49 samples were from out-patient cases (2 samples were positive for Clostridium difficile) and 162 samples were from in-patient cases (19 samples were positive for Clostridium difficile). Clostridium difficile is the frequent pathogenic agent causing diarrheal disease among hospitalized pediatric patients. Development of Clostridium difficile related diarrhea is associated with the antibiotic treatment of pediatric patients hospitalized due to different clinical diagnosis. Targeted early detection of these pathogens is important for the optimal management of diarrheal infection in pediatric patient which will lead to the better clinical outcome and reduction of morbidity rate among hospitalized pediatric patients.

Detection of natural occurrence of Tritrichomonas foetus in cats in Araçatuba, São Paulo, Brazil / Detecção da ocorrência natural de Tritrichomonas foetus em gatos em Araçatuba, São Paulo

The aim of this study was to investigate the occurrence of Tritrichomonas foetus in cats in the area surrounding the city of Araçatuba municipality, State of São Paulo, Brazil. Fecal samples from 129 cats were collected by rectal flush technique. It was compared two diagnosis methods, direct examination of feces and PCR. The presence of T. foetus DNA was verified using PCR by amplification of 347-bp fragment from the primers TFR3 and TFR4 and amplicons of positive cases were sequenced. Statistical analyses were performed investigating the associations between T. foetus infection with gender, age, breed, presence of diarrhea and/or history of diarrhea, previous treatment, lifestyle, origin, environment, and co-infection. T. foetus was observed in one sample (n=129) by direct microscopic examination of feces while PCR was positive in five samples (3.9%). Giardia species and Cryptosporidium species co-infection was also observed. Statistical analyses showed no significant associations between T. foetus infection and all listed factors, although most positive cats were asymptomatic and lived in multi-cat household. The isolates of T. foetus showed 100% identical sequences with other T. foetus isolates from cats around the world. So, the occurrence of T. foetus was confirmed in cats in Araçatuba city (Brazil). This parasite must be considered as a differential diagnosis in cats with diarrhea and also in asymptomatic carriers as source of infection in multi-cat environments.(AU)

O objetivo deste estudo foi investigar a ocorrência de Tritrichomonas foetus em gatos na região do município de Araçatuba, SP, Brasil. Foram coletadas amostras fecais de 129 gatos através da técnica de lavado retal. Dois métodos diagnósticos foram comparados, o exame direto das fezes e a PCR. A presença de DNA de T. foetus foi verificada por meio da PCR através da amplificação de 347 pares de bases a partir dos iniciadores específicos TFR3 e TFR4. Posteriormente, os resultados amplificados das amostras positivas foram sequenciadas. Também foi feita análise estatística a fim de investigar a correlação entre infecção por T. foetus e sexo, idade, raça, presença e/ou histórico de diarreia, tratamento prévio, coinfecção, estilo de vida, origem e tipo de ambiente. O protozoário pôde ser observado em uma amostra através do exame direto das fezes e à PCR foram detectadas cinco amostras positivas (3.9%). Foram detectadas coinfecções por Giardia spp. e Cryptosporidium spp. Não foram observadas correlações entre infecção por T. foetus e todos os fatores listados anteriormente, embora a maioria dos felinos positivos fossem assintomáticos e vivessem em ambientes multigatos. O resultado do sequenciamento genético dos isolados das amostras positivas mostrou 100% de similaridade com outros isolados de felinos no mundo. Assim, a ocorrência de T. foetus foi confirmada em gatos em Araçatuba, São Paulo, Brasil. Sendo assim, o parasito deve considerado como diagnóstico diferencial em gatos com diarreia assim como em portadores assintomáticos como fontes de infecção em ambientes multigatos.(AU)

[Concordance between the zinc sulphate flotation and centrifugal sedimentation methods for the diagnosis of intestinal parasites].

INTRODUCTION: The diagnosis of intestinal parasitic infections depends on the parasite load, the specific gravity density of the parasite eggs, oocysts or cysts, and the density and viscosity of flotation or sedimentation medium where faeces are processed. OBJECTIVE: To evaluate the concordance between zinc sulphate flotation and centrifugal sedimentation in the recovery of parasites in faecal samples of children. MATERIALS AND METHODS: Faecal samples of 330 children from day care centers were evaluated by zinc sulphate flotation and centrifugal sedimentation techniques. The frequencies of detection of parasites by each method were determined and the agreement between the diagnostic techniques was evaluated using the kappa index, with 95% confidence intervals. RESULTS: The faecal flotation in zinc sulphate diagnosed significantly more cases of Trichuris trichiura infection when compared to centrifugal sedimentation (39/330; 11.8% vs. 13/330; 3.9%, p<0.001), with low diagnostic concordance between methods (kappa=0.264; 95% CI: 0.102-0.427). Moreover, all positive samples for Enterobius vermicularis eggs (n=5) and Strongyloides stercoralis larvae (n=3) were diagnosed only by zinc sulphate. No statistical differences were observed between methods for protozoa identification. CONCLUSIONS: The results showed that centrifugal flotation in zinc sulphate solution was significantly more likely to detect light helminths eggs such as those of T. trichiura and E. vermicularis in faeces than the centrifugal sedimentation process.

Concordance between the zinc sulphate flotation and centrifugal sedimentation methods for the diagnosis of intestinal parasites / Concordancia entre los métodos de flotación con sulfato de zinc y sedimentación centrífuga para el diagnóstico de parásitos intestinales

Resumen Introduction: The diagnosis of intestinal parasitic infections depends on the parasite load, the specific gravity density of the parasite eggs, oocysts or cysts, and the density and viscosity of flotation or sedimentation medium where faeces are processed. Objective: To evaluate the concordance between zinc sulphate flotation and centrifugal sedimentation in the recovery of parasites in faecal samples of children. Materials and methods: Faecal samples of 330 children from day care centers were evaluated by zinc sulphate flotation and centrifugal sedimentation techniques. The frequencies of detection of parasites by each method were determined and the agreement between the diagnostic techniques was evaluated using the kappa index, with 95% confidence intervals. Results: The faecal flotation in zinc sulphate diagnosed significantly more cases of Trichuris trichiura infection when compared to centrifugal sedimentation (39/330; 11.8% vs. 13/330; 3.9%, p<0.001), with low diagnostic concordance between methods (kappa=0.264; 95% CI: 0.102-0.427). Moreover, all positive samples for Enterobius vermicularis eggs (n=5) and Strongyloides stercoralis larvae (n=3) were diagnosed only by zinc sulphate. No statistical differences were observed between methods for protozoa identification. Conclusions: The results showed that centrifugal flotation in zinc sulphate solution was significantly more likely to detect light helminths eggs such as those of T. trichiura and E. vermicularis in faeces than the centrifugal sedimentation process.

Abstract Introducción. El diagnóstico de infecciones parasitarias intestinales depende de la carga de parásitos, la densidad de la gravedad específica de los huevos, ooquistes o quistes de parásitos, y de la densidad y viscosidad de los reactivos de flotación o sedimentación usados para procesar las heces. Objetivo. Evaluar la concordancia entre el método de flotación de sulfato de zinc y la sedimentación por centrifugación en la recuperación de parásitos en muestras fecales de niños. Materiales y métodos. Se evaluaron las muestras fecales de 330 niños de guarderías mediante las técnicas de flotación con sulfato de zinc y de sedimentación por centrifugación. Se determinó la frecuencia de detección de parásitos con cada método y se evaluó la concordancia entre las técnicas de diagnóstico mediante el índice kappa, con intervalos de confianza del 95 %. Resultados. Mediante la flotación fecal con sulfato de zinc, se diagnosticó un número significativamente mayor de casos de infección por Trichuris trichiura que con la sedimentación por centrifugación (39/330; 11,8 % Vs. 13/330; 3,9 %) (p<0,001), con poco acuerdo entre los métodos (kappa=0,264; IC95% 0,102-0,427). Además, todas las muestras positivas para huevos de Enterobius vermicularis (n=5) y larvas de Strongyloides stercoralis (n=3) se diagnosticaron solamente por sulfato de zinc. No se observaron diferencias estadísticamente significativas entre los métodos para la identificación de protozoos. Conclusiones. La flotación centrífuga en una solución de sulfato de zinc presentó una probabilidad significativamente mayor de detectar los huevos livianos de helmintos como T. trichiura y E. vermicularis en heces, que el proceso de sedimentación por centrifugación.

Asymptomatic Effluent Protozoa Colonization in Peritoneal Dialysis Patients.

Currently, chronic kidney disease (CKD) is a global health problem. Considering the impaired immunity of CKD patients, the relevance of infection in peritoneal dialysis (PD), and the increased prevalence of parasites in CKD patients, protozoa colonization was evaluated in PD effluent from CKD patients undergoing PD. Overnight PD effluent was obtained from 49 asymptomatic stable PD patients. Protozoa analysis was performed microscopically by searching cysts and trophozoites in direct wet mount of PD effluent and after staining smears. Protozoa were found in PD effluent of 10.2% of evaluated PD patients, namely Blastocystis hominis, in 2 patients, and Entamoeba sp., Giardia sp., and Endolimax nana in the other 3 patients, respectively. None of these patients presented clinical signs or symptoms of peritonitis at the time of protozoa screening. Our results demonstrate that PD effluent may be susceptible to asymptomatic protozoa colonization. The clinical impact of this finding should be further investigated.

DETECTION OF INTESTINAL PARASITES IN THE ENVIRONMENTS OF A PUBLIC SCHOOL IN THE TOWN OF DIAMANTINA , MINAS GERAIS STATE, BRAZIL.

Intestinal parasites are a major public health problem in developing countries, most prevalent in areas where sanitation is poor and the population's hygiene is inadequate. They affect people of all ages, although school-age children are the most susceptible. In this study, we investigated the presence of intestinal parasites in the shared environments of a public school in the town of Diamantina, Minas Gerais State. From December 2012 to February 2013, samples were collected for three months (once a month) by using the Graham method (1941), in duplicate, by affixing a 6 x 5 cm clear tape, six times in each collection site, in a space of about 30 cm2. Then, each tape was positioned longitudinally on a microscope slide and the identification of the biological forms of the parasites was performed with the aid of a 40X objective from an optical microscope. Eleven sites were selected for sampling. Cysts of Entamoeba coli were the most frequently found in this study (50%), followed by Hymenolepis diminuta eggs (27.6%), Iodamoeba butschllii cysts (5.6%), Ascaris lumbricoides eggs (5.6%), Taenia species eggs (5.6%) and hookworm eggs (5.6%). The highest positivity rates were found in the samples drawn from the cafeteria's eating table. The results have indicated the need to improve the cleaning in the school environments, as well as the development of educational practices that may help in the preservation of public health.

Perkinsus beihaiensis infecting the oyster Crassostrea rhizophorae under cultivation and in natural stock in Camamu Bay, Bahia, Brazil / Perkinsus beihaiensis infectando a ostra Crassostrea rhizophorae em cultivo e em estoque natural na Baía de Camamu, Bahia, Brasil

This study investigated the infection of Perkinsus beihaiensis (Perkinsozoa) in the oyster Crassostrea rhizophorae, both from a long-line cultivation system and from a nearby intertidal zone of mangrove, both in the state of Bahia, northeastern Brazil. The collections were performed in October and November 2012, and in January 2013. The oysters (n = 300) were measured, examined macroscopically for signs of infection and then submitted to the following laboratory techniques: histology, Ray"s fluid thioglycollate medium assay (RFTM), polymerase chain reaction (PCR) and sequencing, which confirmed the identification of the pathogen. Histological and RFTM analyses showed, respectively, a mean prevalence of 93.3% and of 69%. The infection was usually mild or very mild. There was no significant difference (p > 0.05) between the environments in terms of infection prevalence or severity. This is the first record of P. beihaiensis in the state of Bahia and the second in oysters from Brazil and South America...

Foi investigada a infecção de Perkinsus beihaiensis (Perkinsozoa) na ostra Crassostrea rhizophorae em um sistema de cultivo do tipo espinhel e em um estoque natural de ostras no manguezal adjacente ambos localizados no estado da Bahia, Nordeste do Brasil. As colheitas foram realizadas em outubro e novembro de 2012 e em janeiro de 2013. As ostras (n = 300) foram medidas, examinadas macroscopicamente quanto a sinais da infecção e submetidas às técnicas laboratoriais: histologia, ensaio em meio de cultivo de tioglicolato de Ray (RFTM), reação em cadeia da polimerase (PCR) e sequenciamento, que confirmou a identificação do patógeno. As análises histológicas e o RFTM mostraram, respectivamente, prevalência média de 93,3% e de 69%. A infecção foi geralmente leve ou muito leve. Não houve diferença significativa (p > 0,05) entre os ambientes em termos de prevalência ou severidade da infecção. Este é o primeiro registro de P. beihaiensis no estado da Bahia e o segundo em ostras do Brasil e América do Sul...