Survey of vector-borne and nematode parasites involved in the etiology of anemic syndrome in sheep from Southern Brazil / Pesquisa de hemoparasitos transmitidos por vetores e nematódeos envolvidos na etiologia de síndrome anêmica em ovinos no Sul do Brasil

Abstract Although anemia has been historically linked to Haemonchus contortus infection, other infectious agents, such as hemotropic mycoplasmas and tick-borne disease pathogens, may also lead to anemic crisis in sheep. This study has aimed to investigate infections related to anemia in a sheep herd from Bandeirantes City, Paraná State, southern Brazil. Seven out of forty-two (16.6%; 95% CI: 8.32-30.6%) sheep were positive for hemoplasmas by a PCR targeting the 16S rRNA gene and all tested negative for A. marginale/A. ovis and Babesia/Theileria spp. by PCR based on msp4 and 18S rRNA genes, respectively. Two (4.7%; 95% CI: 1.32-15.79%) animals were infested with Rhipicephalus microplus ticks. Fecal egg counting was performed in 38 sheep and 24 (63.15%; 95% CI: 47.2-76.6%) presented > 500 eggs per gram. Phylogenetic analysis of partial sequences of the detected hemotropic Mycoplasma sp. 16S and 23S rRNA genes confirmed that the animals were infected with Mycoplasma ovis. Polymorphism analysis of partial 16S rRNA sequences showed three different genotypes of M. ovis infecting sheep assessed in the present study. Mycoplasma ovis and gastrointestinal nematodes occurs in sheep from the northern region of Paraná State.

Resumo Embora a principal causa de anemia seja historicamente relacionada à infecção por Haemonchus contortus, outros agentes infecciosos, como micoplasmas hemotrópicos e patógenos transmitidos por carrapatos, também podem causar quadros anêmicos em ovinos. O presente estudo objetivou investigar infecções relacionadas à anemia em um rebanho de ovinos, na cidade de Bandeirantes, Estado do Paraná, sul do Brasil. Sete (16,6%; 95% CI: 8,32-30,6%) de 42 ovinos foram positivos para hemoplasmas pela PCR do gene 16S rRNA, enquanto todos foram negativos para A. marginale/A. ovis e Babesia/Theileria spp. por ensaios da PCR baseados nos genes msp4 e 18S rRNA, respectivamente. Dois (4,7%; 95% CI: 1,32-15,79%) animais estavam infestados por carrapatos Rhipicephalus microplus. Dos 38 animais nos quais foi realizada a contagem de ovos por grama de fezes (OPG), 24 (63,15%; 95% CI: 47,2-76,6%) apresentaram valores >500 para OPG. A análise filogenética das sequências parciais dos genes 16S rRNA e 23S rRNA de hemoplasmas confirmou a infecção por Mycoplasma ovis. A análise de polimorfismos de um fragmento do gene 16S rRNA mostrou a ocorrência de três genótipos diferentes de M. ovis nos animais. Mycoplasma ovis e nematódeos gastrointestinais ocorrem em ovinos da região nordeste do Estado do Paraná.

Molecular detection of vector borne pathogens in anemic and thrombocytopenic dogs in southern Brazil / Detecção molecular de patógenos transmitidos por vetores em cães anemicos e trombocitopenicos no Sul do Brasil

Abstract Arthropod-borne pathogens are medically important because of their ability to cause diseases in their hosts. The purpose of this study was to detect the occurrence of Ehrlichia spp., piroplasmids and Hepatozoon spp. in dogs with anemia and thrombocytopenia in southern Brazil. EDTA-whole blood was collected from 75 domestic dogs presenting anemia or/and thrombocytopenia from Guarapuava, state of Paraná, Brazil. DNA samples were subjected to conventional PCR assays for Ehrlichia spp. (dsb), piroplasmids (18S rRNA) and Hepatozoon spp. (18S rRNA), followed by sequencing and phylogenetic analyses. Among the 75 dogs, one (1.33%) was positive for Hepatozoon sp. and six (8%) were positive for piroplasmids in 18S rRNA cPCR assays. None of the dogs showed positive results in Ehrlichia spp.-cPCR targeting dsb gene. The phylogenetic analyses revealed that three piroplasm sequences were clustered with Rangellia vitalii, while one sequence was grouped with B. vogeli. The only sequence obtained from Hepatozoon spp.-PCR protocol was pooled with H. canis. Therefore, there is urgent need for differential molecular diagnosis of the two piroplasm species cited as etiological agents in clinical cases of canine hemoparasitic diseases, given the higher pathogenic potential of R. vitalii than of B. vogeli.

Resumo Agentes transmitidos por artrópodes têm grande importância na medicina veterinária devido à sua capacidade de causar doenças graves em seus hospedeiros. O presente estudo objetivou investigar a ocorrência de três patógenos transmitidos por vetores, Ehrlichia canis, Rangelia vitalii e Hepatozoon canis, em cães na região sul do Brasil. Foram coletadas amostras de sangue total de 75 cães domésticos que apresentavam anemia e/ou trombocitopenia, em Guarapuava, Paraná, Brasil. As amostras de DNA foram submetidas à técnica de PCR convencional para E. canis (dsb), piroplasmídeos (18S rRNA) e Hepatozoon spp. (18S rRNA), seguida de sequenciamento e análises filogenéticas. Das 75 amostras, uma (1,33%) foi positiva para Hepatozoon spp. e seis (8%) foram positivas para Babesia spp. Nenhuma amostra mostrou resultados positivos para Ehrlichia spp. utilizando a detecção pelo gene dsb. As análises filogenéticas revelaram que três sequências obtidas foram agrupadas no mesmo clado que R. vitalii , enquanto uma foi agrupada juntamente com B. vogeli. A única sequência obtida pelo protocolo de PCR para Hepatozoon spp. foi agrupada juntamente com H. canis. Assim, é justificada necessidade de diferenciação das espécies de piroplasmas, através do diagnóstico molecular, como agentes etiológicos nos casos clínicos de hemoparasitose canina, considerando o potencial patogênico de R. vitalii quando comparado à B. vogeli.

Anemia crónica grave por Ancylostoma duodenale en Ecuador: diagnóstico por duodenoscopia / Chronic and severe anemia caused by Ancylostoma duodenale in Ecuador: diagnosis by duodenoscopy

Resumen Durante 11 años un varón de 38 años de edad, residente en una región subtropical de Ecuador, había sido diagnosticado de anemia crónica y tratado con transfusiones de sangre en un hospital de la provincia de Cotopaxi, Ecuador. Fue transferido a Quito por una anemia grave, con hemoglobina de 4 g/dL. Se realizó una duodenoscopia en que se observaron nemátodos adultos, identificados posteriormente como Ancylostoma duodenale. El paciente fue tratado exitosamente con albendazol durante cinco días consecutivos y transfusiones de sangre. En el seguimiento a los ocho meses, no se encontró anemia ni huevos de anquilostomas en el examen de heces.

For 11 years, a 38-year-old male residing in a subtropical region of Ecuador, was repeatedly diagnosed with chronic anemia, and treated with blood transfusions in a hospital of province of Cotopaxi, Ecuador. He was transferred to Quito for severe anemia, having hemoglobin of 4 g/dL. Duodenoscopy was performed and adult nematodes, identified later as Ancylostoma duodenale, were observed. The patient was successfully treated with albendazole for five consecutive days and given blood transfusions. In the control visit at eight months, without anemia and no hookworm ova in the stool examined were found.

Influence of age on the haemoglobin concentration of malaria-infected patients in a reference centre in the Brazilian Amazon

Anaemia is amongst the major complications of malaria, a major public health problem in the Amazon Region in Latin America. We examined the haemoglobin (Hb) concentrations of malaria-infected patients and compared it to that of malaria-negative febrile patients and afebrile controls. The haematological parameters of febrile patients who had a thick-blood-smear performed at an infectious diseases reference centre of the Brazilian Amazon between December 2009-January 2012 were retrieved together with clinical data. An afebrile community control group was composed from a survey performed in a malaria-endemic area. Hb concentrations and anaemia prevalence were analysed according to clinical-epidemiological status and demographic characteristics. In total, 7,831 observations were included. Patients with Plasmodium falciparum infection had lower mean Hb concentrations (10.5 g/dL) followed by P. vivax-infected individuals (12.4 g/dL), community controls (12.8 g/dL) and malaria-negative febrile patients (13.1 g/dL) (p < 0.001). Age, gender and clinical-epidemiological status were strong independent predictors for both outcomes. Amongst malaria-infected individuals, women in the reproductive age had considerably lower Hb concentrations. In this moderate transmission intensity setting, both vivax and falciparum malaria are associated with reduced Hb concentrations and risk of anaemia throughout a wide age range.

The effect of hookworm infection and urinary scistosomiasis on blood hemoglobiin concentration of schoolchildren living in northern Mozambique / Infecções por ancilostomídeos e Schistosoma haematobium e sua correlação com a concentração sanguínea de hemoglobina em crianças moçambicanas

This study aims to assess the association between schistosomiasis and hookworm infection with hemoglobin levels of schoolchildren in northern Mozambique. Through a cross-sectional survey, 1,015 children from five to 12 years old in the provinces of Nampula, Cabo Delgado and Niassa were studied. Hookworm infection and urinary schistosomiasis were diagnosed, through Ritchie and filtration methods, with a prevalence of 31.3% and 59.1%, respectively. Hemoglobin levels were obtained with a portable photometer (Hemocue®). The average hemoglobin concentration was 10.8 ± 1.42 g/dL, and 62.1% of the children presented levels below 11.5 g/dL, of which 11.8% of the total number of children had hemoglobin levels below 9 g/dL. A multiple linear regression analysis demonstrated negative interactions between hemoglobin levels and ancylostomiasis, this being restricted to the province of Cabo Delgado (β = -0.55; p < 0.001) where an independent interaction between hemoglobin levels and urinary schistosomiasis was also observed (β = -0.35; p = 0.016). The logistical regression model indicated that hookworm infection represents a predictor of mild (OR = 1.87; 95% CI = 1.17-3.00) and moderate/severe anemia (OR = 2.71; 95% CI = 1.50 - 4.89). We concluded that, in the province of Cabo Delgado, hookworm and Schistosoma haematobium infections negatively influence hemoglobin levels in schoolchildren. Periodical deworming should be considered in the region. Health education and improvements in sanitary infrastructure could achieve long-term and sustainable reductions in soil-transmitted helminthiases and schistosomiasis prevalence rates.

Este estudo tem como objetivo avaliar a relação entre a ancilostomíase e a esquistossomíase urinária com as concentrações sanguíneas de hemoglobina em crianças escolares no norte de Moçambique. Em estudo transversal, 1.015 crianças com idade entre cinco e 12 anos foram incluídas, nas Províncias de Nampula, Cabo Delgado e Niassa. A ancilostomíase e a esquistossomíase urinária foram diagnosticadas através das técnicas de Ritchie e de filtração da urina, respectivamente; prevalências de 31,3% e 59,1% foram observadas. As concentrações sanguíneas de hemoglobina foram obtidas com um fotômetro portátil (Hemocue). A concentração média de hemoglobina foi 10,8 ± 1.42 g/dL, 62,1% das crianças apresentaram concentração abaixo de 11,5 g/dL e 11,8% apresentaram nível abaixo de 9 g/dL. A regressão linear múltipla demonstrou interações negativas entre os níveis de hemoglobina e i) a infecção por ancilostomídeos (β = -0,55; p < 0,001) e ii) a esquistossomíase urinária (β = -0,35; p = 0,016), ambas associações restritas à Província de Cabo Delgado. Também em Cabo Delgado, o modelo de regressão logística demonstrou que a infecção por ancilostomídeos representa um preditor de anemia leve (OR = 1,87; 95% CI = 1,17-3,00) e anemia moderada/grave (OR = 2,71; 95% CI = 1,50 - 4,89). O estudo conclui que em Cabo Delgado, Moçambique, as infecções por ancilostomídeos e Schistosoma haematobium estão significativamente associadas a uma menor concentração sanguínea de hemoglobina em crianças em idade escolar. A administração periódica de anti-helmínticos deve ser feita regularmente. Melhorias na infraestrutura sanitária das regiões estudadas são as medidas mais eficazes para controle destas parasitoses.

Apoptosis of non-parasitised red blood cells in Plasmodium yoelii malaria

Recently, while studying erythrocytic apoptosis during Plasmodium yoelii infection, we observed an increase in the levels of non-parasitised red blood cell (nRBC) apoptosis, which could be related to malarial anaemia. Therefore, in the present study, we attempted to investigate whether nRBC apoptosis is associated with the peripheral RBC count, parasite load or immune response. To this end, BALB/c mice were infected with P. yoelii 17XL and nRBC apoptosis, number of peripheral RBCs, parasitaemia and plasmatic levels of cytokines, nitric oxide and anti-RBC antibodies were evaluated at the early and late stages of anaemia. The apoptosis of nRBCs increased at the late stage and was associated with parasitaemia, but not with the intensity of the immune response. The increased percentage of nRBC apoptosis that was observed when anaemia was accentuated was not related to a reduction in peripheral RBCs. We conclude that nRBC apoptosis in P. yoelii malaria appears to be induced in response to a high parasite load. Further studies on malaria models in which acute anaemia develops during low parasitaemia are needed to identify the potential pathogenic role of nRBC apoptosis.

Enhanced T cell activation in Plasmodium falciparum malaria-infected human immunodeficiency virus-1 patients from Mozambique

Human immunodeficiency virus (HIV)-1 infection has an important impact on malaria. Plasmodium falciparum and HIV-1 co-infected patients (Pf/HIV) present with a high degree of anaemia, enhanced parasitaemia and decreased CD4+ T cell counts, which increase the risk of developing severe malaria. In addition, infection with either Pf or HIV-1 alone causes extensive immune activation. Our hypothesis was that lymphocyte activation is potentiated in Pf/HIV co-infected patients, consequently worsening their immunosuppressed state. To test this hypothesis, 22 Pf/HIV patients, 34 malaria patients, 29 HIV/AIDS patients and 10 healthy controls without malaria or HIV/acquired immune deficiency syndrome (AIDS) from Maputo/Mozambique were recruited for this study. As expected, anaemia was most prevalent in the Pf/HIV group. A significant variation in parasite density was observed in the Pf/HIV co-infected group (110-75,000 parasites/µL), although the median values were similar to those of the malaria only patients. The CD4+ T cell counts were significantly lower in the Pf/HIV group than in the HIV/AIDS only or malaria only patients. Lymphocyte activation was evaluated by the percentage of activation-associated molecules [CD38 expression on CD8+ and human leukocyte antigen-DR expression on CD3+ T cells]. The highest CD38 expression was detected in the Pf/HIV co-infected patients (median = 78.2%). The malaria only (median = 50%) and HIV/AIDS only (median = 52%) patients also exhibited elevated levels of these molecules, although the values were lower than those of the Pf/HIV co-infected cases. Our findings suggest that enhanced T-cell activation in co-infected patients can worsen the immune response to both diseases.

Improving the production of the denatured recombinant N-terminal domain of rhoptry-associated protein 2 from a Plasmodium falciparum target in the pathology of anemia in falciparum malaria

Rhoptry-associated protein 2 (RAP2) is known to be discharged from rhoptry onto the membrane surface of infected and uninfected erythrocytes (UEs) ex vivo and in vitro and this information provides new insights into the understanding of the pathology of severe anemia in falciparum malaria. In this study, a hexahistidine-tagged recombinant protein corresponding to residues 5-190 of the N-terminal of Plasmodium falciparum RAP2 (rN-RAP2) was produced using a new method of solubilization and purification. Expression was induced with D-lactose, a less expensive alternative inducer to the more common isopropyl-²-D-thio-galactopyranosidase. The recombinant protein was purified using two types of commercially-available affinity columns, iminodiacetic and nitrilotriacetic. rN-RAP2 had immunogenic potential, since it induced high titers of anti-RAP2 antibodies in mice. These antibodies recognized full-length RAP2 prepared from Triton X-100 extracts from two strains of P. falciparum. In fact, the antibody recognized a 29-kDa product of RAP2 cleavage as well as 82 and 70-kDa products of RAP1 cleavage. These results indicate that the two antigens share sequence epitopes. Our expressed protein fragment was shown to contain a functional epitope that is also present in rhoptry-derived ring surface protein 2 which attaches to the surface of both infected and UEs and erythroid precursor cells in the bone marrow of malaria patients. Serum from malaria patients who developed anemia during infection recognized rN-RAP2, suggesting that this protein fragment may be important for epidemiological studies investigating whether immune responses to RAP2 exacerbate hemolysis in falciparum malaria patients.

Bilateral epistaxis after face washing in a pond in a two year old child

We report a 2 year old boy with epistaxis, melena, fever and anaemia. In an otolaryngologist consultation a leech was seen in the nasopharynx. His face was washed in a pond 3 days prior to admission

Giardiasis and its relation to anemia and failure to growth in children

Giardiasis is one of the most famous parasitological infections in the humans, the causing factor is the flagellate protozoan Giardia Lamblia which is living in the upper part of the small intestine attaching to the mucousa membrane. The infection with this flagellate may cause diarrhea disease that varies in severity, wherease the chronic infection with this parasite may cause mal absorption and a celiac disease like syndrome which lead specially in children to anemia and retardation in growth and development. Our studying performed on four groups, which contained 465 child. We diagnosed the giardia parasite through fecal microscopic test, and we applied hematological tests and special measurement to diagnose the failure to growth. We found that the percentage of giardiasis is 24%, and the percentage of children with anemia beside giardiasis is 37%, while the percentage of children with failure to growth and giardiasis is 43%, but the percentage of children with anemia and failure to growth and giardiasis is 22%

Parasitosis intestinal y anemia en escolares de zonas aledañas al río Paraguay en Asunción / Annual reports 1997 / Intestinal parasitosis and anemia in school aged children from areas around the Paraguay river in Asunción

El presente trabajo ha tenido como objetivo establecer la situación actual de la anemia parasito-nutricional en niños de 6 a 10 años que concurren a escuelas aledañas al Río Paraguay en Asunción. Fueron estudiados 220 niños según muestreo no probabilístico de conveniencia, en quienes se evaluaron edad, sexo, peso, talla, hemoglobina, hematocrito, glóbulos rojos, índices hematimétricos (VCM, HCM y CHCM), sideremia y parásitos intestinales. En la población total estudiada (50.5 por ciento del sexo femenino y 49.5 por ciento del sexo masculino) se observó 76.4 por ciento de parasitosis intestinal, 11.8 por ciento de anemia, 5 por ciento de déficit de hierro sérico y 2.7 por ciento de desnutrición. Se presentan, además las prevalencias de parásitos en niños anémicos y no anémicos. En esta población la anemia no está significativamente asociada con la parasitosis intestinal, y siendo las anemias normocíticas y no ferroprivas las más frecuentemente observadas, se sugiere que otras etiologías diferentes a la parasitosis (no evaluadas en este trabajo) sean causantes de las mismas