New species of Polystomoides (Monogenoidea: Polystomatidae) parasitizing the urinary bladder of a freshwater turtle in Brazil.

Trachemys dorbigni is the most abundant freshwater turtle species in Rio Grande do Sul, southern Brazil. Chelonians are known to host a wide variety of pathogens, including viruses, bacteria, hemoparasites and helminths. Among these, nine genera of polystomatid flatworms (Monogenoidea; Polystomatidae) infect freshwater turtles: Apaloneotrema, Aussietrema, Fornixtrema, Manotrema, Pleurodirotrema, Polystomoidella, Polystomoides, Uropolystomoides and Uteropolystomoides. However, little is known about the biology of these parasites in the Neotropical Realm. Through investigative cystoscopy, specimens of Polystomatidae were located inside the urinary bladder of the host T. dorbigni. Retrieved specimens were fixed and stained whole mounts prepared for taxonomic identification. In the present paper, a new species of Polystomoides (Monogenoidea: Polystomatidae) parasitizing the urinary bladder of a freshwater turtle of the species T. dorbigni in Brazil is described. Polystomoides santamariensis n. sp. differs from the congeneric species on the length of the genital spines, which are longer. Given the enormous diversity of freshwater turtles around the world, it is likely that a large number of chelonian polystomatids are still unknown.

Eucalyptus oil nanoemulsions against eggs and larvae of Haemonchus contortus.

Haemonchus contortus is a highly pathogenic and prevalent helminth that causes many deaths in sheep herds. Anthelmintics are usually employed to overcome this issue; however, they do not guarantee immediate and lasting efficacy because of the occurrence of drug-resistant parasites. Among substances that are used in scientific studies for parasitic control, essential oils are known to have different pharmacological properties. However, they demonstrate instability owing to several factors, and therefore, nanoemulsification is considered an alternative to control the instability and degradability of these compounds. The objective of this study was to evaluate the cytotoxicity of nanoemulsions containing essential oil of Eucalyptus globulus against the blood of healthy sheep and to verify their activity against the parasite H. contortus in sheep. The results presented adequate nanotechnological characteristics (diameter 72 nm, PDI 0.2, zeta -11 mV, and acidic pH) and adequate morphology. Further, the corona effect and cytotoxic profiles of the free oil and nanoemulsion against blood cells from healthy sheep were evaluated. The tests results did not present a toxicity profile. For evaluating efficacy, we observed an important anthelmintic action of the nanoemulsion containing oil in comparison to the free oil; the results demonstrate a potential role of the nanoemulsion in the inhibition of egg hatchability and the development of larvae L1 to L3 (infective stage). Based on these results, we developed an important and potential anthelmintic alternative for the control of the parasite H. contortus.

Involvement of ectonucleotidases and purinergic receptor expression during acute Chagas disease in the cortex of mice treated with resveratrol and benznidazole.

Chagas disease (CD) is caused by the parasite Trypanosoma cruzi. CD affects people worldwide, primarily in tropical areas. The central nervous system (CNS) is an essential site for T. cruzi persistence during infection. The protozoan may pass through the blood-brain barrier and may cause motor and cognitive neuronal damage. Once in the CNS, T. cruzi triggers immune responses that the purinergic system can regulate. Treatment for CD is based on benznidazole (BNZ); however, this agent has negative side-effects and is toxic to the host. For this reason, we investigated whether resveratrol (RSV), a potent antioxidant and neuroprotective molecule, would modulate purinergic signaling and RSV alone or in combination with BNZ would prevent changes in purinergic signaling and oxidative damage caused by T. cruzi. We infected mice with T. cruzi and treated them with RSV or BNZ for 8 days. Increases in ATP and ADP hydrolysis by NTPDase in the total cortex of infected animals were observed. The treatment with RSV in infected group diminished ATP, ADP, and AMP hydrolysis compared to infected group. The combination of RSV + BNZ decreased AMP hydrolysis in infected animals compared to the INF group, exerting an anti-inflammatory effect. RSV acted as a neuroprotector, decreasing adenosine levels. Infected animals presented an increase of P2X7 and A2A density of purine receptors. RSV reduced P2X7 and A2A and increased A1 density receptors in infected animals. In addition, infected animals showed higher TBARS and reactive oxygen species (ROS) levels than control. RSV diminished ROS levels in infected mice, possibly due to antioxidant properties. In short, we conclude that resveratrol could act as a neuroprotective molecule, probably preventing inflammatory changes caused by infection by T. cruzi, even though the mice experienced high levels of parasitemia.

Resveratrol impacts in oxidative stress in liver during Trypanosoma cruzi infection.

Trypanosoma cruzi is the causative agent of Chagas disease, infecting the heart, intestines and liver tissues. There is growing evidence that oxidative stress, defined as a persistent imbalance between highly oxidative compounds and antioxidant defenses, is a marker of tissue inflammation; it is related to immune responses such as damage, as well as to strand breaks in DNA contributing to disease progression. Antioxidant agents help mitigate the damage caused by inflammation, preventing or slowing damage to cells caused by free radicals. In this sense, resveratrol (RSV) is an important polyphenol that demonstrates antioxidant effects. It reverses damage caused by several infectious diseases. The aim of the present study was to determine whether treatment with RSV would prevent or minimize oxidative damage caused by T. cruzi. The animals were divided into four groups (n = 5): A) control; B) control + RSV; C) infected and D) infected + RSV. The infected groups received 1 x 104 Y strain trypomastigotes via intraperitoneal injection; after confirmation of infection, the mice received RSV 100 mg/kg for seven days orally. On the 8th day post-infection, we collected liver tissue for analysis of oxidant/antioxidant status: superoxide dismutase (SOD), catalase (CAT), and glutathione s-transferase (GST) activities, as well as reactive oxygen species (ROS), non-protein thiols (NPSH), thiols, carbonyl protein, thiobarbituric acid reactive substance (TBARS), and finally, the nitrite/nitrate ratio (NOx) levels were determined. The administration of RSV did not exert direct effect on parasitemia. The infection produced high levels of TBARS, NOx, and ROS levels in liver tissue, suggesting cellular injury with production of free radicals in animals infected by T. cruzi. RSV positively modulated SOD and aumenting GST activities enzymes in infected animals. Protein thiols levels in infected animals were lower than those of control. Taken together, the data suggest T. cruzi causes hepatic oxidative stress, and RSV 100 mg/kg for seven days it's dosen't seem minimized these negative effects in the acute phase of disease.

Adulticidal Activity of Melaleuca alternifolia (Myrtales: Myrtaceae) Essential Oil With High 1,8-Cineole Content Against Stable Flies (Diptera: Muscidae).

The stable fly, Stomoxys calcitrans (Linnaeus 1758), is a hematophagous fly responsible for causing loss of performance in horses, causing losses in cattle productivity, and impacting the animals' health through the spread of pathogenic microorganisms. The objective of this work was to investigate the insecticidal activity of essential oil obtained from Melaleuca alternifolia (Cheel), presenting high 1,8-cineole content, against S. calcitrans adults. Insecticidal activity was determined using surface application methods and exposure to oil impregnated paper. It was observed that treatments at 25 and 50 µg/cm2 (P < 0.05) present fumigant activity through exposure to the impregnated paper, and in the first 15 min of exposure, the mortality rates obtained for these treatments were, respectively (96.6 ± 3.3% and 100%), equivalent to the positive control. Using the superficial application method, the only treatment concentration presenting adulticidal action was 5% (w/v) (P < 0.05). Respective toxicities LC50 (%, w/v) and LC80 for the impregnated paper method were 1.06 ± 0.02 and 1.47 ± 0.17; for the superficial application method, they were 3.82 ± 0.65 and 5.53 ± 0.74. As demonstrated, M. alternifolia essential oil presents adulticidal potential against S. calcitrans.

Litomosoides silvai (Nematoda: Onchocercidae) parasitizing Akodon montensis (Rodentia: Cricetidae) in the southern region of Brazil / Litomosoides silvai (Nematoda: Onchocercidae) parasitando Akodon montensis (Rodentia: Cricetidae) na região Sul do Brasil

Abstract In the present study, Litomosoides silvai parasitizing Akodon montensis in the southern region of Brazil is reported for the first time. New morphological information is provided for some structures of this nematode species, such as a flattened cephalic extremity, presence of two dorsal cephalic papillae, female tail with a constriction at its tip, "s" shaped vagina, spicules characteristic of the carinii species group and microfilaria tail constricted at the tip. This nematode was found parasitizing the thoracic cavity with a prevalence of 10% (2/20), mean intensity of 4 (6/2), mean abundance of 0.4 (8/20) and range of infection of 2-6 specimens per host, in southern Brazil. This occurrence of L. silvai in A. montensis is a new geographical record for southern Brazil, in the Upper Paraná Atlantic Forest ecoregion of the northwestern region of Rio Grande do Sul, which is part of the Atlantic Forest biome.

Resumo No presente estudo é relatado pela primeira vez Litomosoides silvai parasitando Akodon montensis coletados na região Sul do Brasil. Foram fornecidas novas informações morfológicas para algumas estruturas desta espécie de nematódeo, tais como extremidade cefálica achatada, a presença de duas papilas cefálicas dorsais, cauda das fêmeas com uma constrição na ponta da cauda, vagina em forma de "s", espículas de característica do grupo de espécies de carinii e cauda da microfilária com constrição na ponta. Este nematódeo parasitava a cavidade torácica com uma prevalência de 10% (2/20), intensidade média de 4 (8/2) e abundância média de 0,4 (8/20), e intervalo de infecção de 2-6 espécimes por hospedeiro no Sul do Brasil. A ocorrência de L. silvai em A. montensis é um novo registro geográfico, no sul do Brasil, a noroeste do estado do Rio Grande do Sul, na ecorregião da Mata Atlântica do Alto Paraná, parte do bioma da Mata Atlântica.

Adapted Bailenger method improves the rate of Ascaris suum eggs recovery from liquid pig manure compost / Adaptações no método de Bailenger melhora taxa de recuperação de ovos de Ascaris suum em dejetos líquidos de suínos compostado

ABSTRACT: Liquid pig manure (LPM) is widely used as a compost fertilizer for vegetable crops destined for human consumption. However, these wastes may contain parasites eggs, such as the nematode Ascaris suum, that pose serious health risks to humans. We attempted to determine the most appropriate technique for recovering A. suum eggs from LPM compost. Samples were collected from two waste sources during composting, including 23 samples containing LPM, sawdust, and wood shavings, and 14 samples of LPM alone-both in triplicate. Samples were analyzed using several different recovery methods. Recovery of eggs by the modified Bailenger method with adaptations was significantly more effective and recovered 57% more eggs than by the modified Bailenger method alone. Willis-Mollay method, modified Faust method, and the simple sedimentation technique only recovered 4.4%, 13.9%, and 26% of eggs, respectively, compared with the modified Bailenger method with adaptations, indicating that the adjustments made to the Bailenger method were key to improving the recovery of A. suum eggs from compost and LPM.

RESUMO: Dejetos líquidos de suínos (DLS) são amplamente utilizado como adubo para as culturas vegetais destinados ao consumo humano. No entanto, estes resíduos podem conter os ovos de parasitas, tais como o nematoide Ascaris suum, que apresentam riscos graves para a saúde dos seres humanos. Neste estudo tentamos determinar a técnica mais apropriada para a recuperação de ovos de A. suum no composto de DLS e nos dejetos líquidos. Foram coletadas amostras de dois tratamentos destes resíduos, que consiste de 23 amostras de DLS + serragem + maravalha e 14 amostras de DLS sozinhos, todos em triplicata. Em ambos, dejetos líquidos e dejetos compostado foram analisados usando um método modificado Bailenger (BM), um método de Faust modificado (FM), o método de Willis-Mollay (WM), sedimentação simples (SS), e pelo método de Bailenger modificado com adaptações (BMA). A recuperação de ovos pelo Método BMA foi significativamente mais eficaz do que pela BM, com 57% mais ovos recuperados usando a técnica de BMA; além disso, o WM, FM, e o método SS recuperaram 4,4%, 13,9% e 26%, respectivamente, em comparação com o método BMA, indicando que os ajustes feitos com o método BM foram fundamentais para melhorar a recuperação de ovos de A. suum ovos em dejetos compostados e DLS puro.

Reemerging of natural infection by Trypanosoma evansi in horses in Arari, Marajó Island, Brazil / Reemergência da infecção natural por Trypanosoma evansi em cavalos, na região do Arari, Ilha do Marajó, Brazil

ABSTRACT: Two outbreaks of trypanosomiasis by Trypanosoma evansi in horses in the municipality of Chaves, Marajó Island, Pará State, Brazil are reported. The first outbreak occurred in April 2011 in a farm with 147 horses; of these, 47 (31.97%) got sick, and 40 (27.21%) died. The second outbreak occurred in May 2012 and involved nine properties. Of a total of 679 horses, 209 (30.07%) got sick, and 183 (26.97%) died. The main clinical signs observed in these horses were weight loss, abdominal edema, toe dragging, and pelvic muscle atrophy. Necropsy was performed in two horses, one from each outbreak. Macroscopic alterations were not seen in the horse from the first outbreak; however, the horse from the second outbreak exhibited emaciation, pallor and icterus, as well as enlarged spleen with prominent white pulp at cut surface. Histopathology of the central nervous system of both animals revealed mild to severe, diffuse lymphoplasmacytic encephalitis, with presence of Mott cells. Immunohistochemistry for T. evansi revealed structures similar to intralesional trypomastigote forms. Between November and December of 2013, an epidemiological survey was carried out in the municipalities of Cachoeira do Arari, Santa Cruz do Arari, Salvaterra, Soure, and Chaves. Only Santa Cruz do Arari and Chaves had reported cases of the disease. Blood samples were collected from 243 horses for detection of T. evansi DNA by polymerase chain reaction (PCR) and 20 were positive.

RESUMO: São relatados dois surtos de tripanossomíase por Trypanosoma evansi , em cavalos no município de Chaves, Ilha do Marajó, Pará, Brasil. O primeiro surto ocorreu em abril de 2011 em uma fazenda com 147 cavalos, em que 47 (31,97%) adoeceram e 40 (27,21%) morreram. O segundo ocorreu em maio de 2012 e envolveu nove propriedades. De um total de 679 cavalos, 209 (30,07%) adoeceram e 183 (26,97%) morreram. Os principais sinais clínicos observados foram perda de peso, edema abdominal, emboletamento dos membros e atrofia da musculatura da região pélvica e membros posteriores. A necropsia foi realizada em dois cavalos, um de cada surto. O equino do primeiro surto não apresentou lesões macroscópicas, enquanto o segundo, do segundo surto, apresentava emagrecimento, palidez e icterícia, aumento do baço com protrusão da polpa branca ao corte. O sistema nervoso central de ambos os animais revelou uma leve à severa encefalite linfoplasmocitária, difusa, com presença de células de Mott. A imunohistoquímica para T. evansi revelou estruturas similares a formas tripomastigotas nas lesões. Entre novembro e dezembro de 2013, um levantamento epidemiológico foi realizado nos municípios de Cachoeira do Arari, Santa Cruz do Arari, Salvaterra, Soure e Chaves. Somente Santa Cruz do Arari e Chaves relataram casos da doença. Foram coletadas amostras de sangue de 243 equinos e realizada a reação em cadeia de polimerase (PCR) para a detecção do DNA de T. evansi das quais 20 foram positivas.

Multiparasitism in a wild cat (Leopardus colocolo) (Carnivora: Felidae) in southern Brazil / Multiparasitismo em um felino silvestre (Leopardus colocolo) (Carnivora: Felidae) no Sul do Brasil

Abstract Parasitic diseases reflect the health and balance of ecosystems, affecting not only individuals but also entire populations or communities. The aim of this study was to report on the diversity of parasitic helminths detected in the feces of a wild feline in southern Brazil. Parasites were obtained from fecal samples, and four techniques were used for parasitological examination: direct examination, centrifugal flotation with zinc sulfate (Faust technique), simple sedimentation (Hoffman technique) and Baermann-Moraes. The parasites were identified through micrometry and morphology, as follows: Ancylostoma sp., Toxocara sp., Trichuridae, Aelurostrongylus abstrusus, Alaria sp., and Spirometra sp. We recorded the genus Ancylostoma parasitizing L. colocolo for the first time.

Resumo Doenças parasitárias refletem a saúde e o equilíbrio dos ecossistemas, influenciando não só um indivíduo e sim uma população ou comunidade. Este trabalho teve por objetivo relatar a diversidade de helmintos encontradas nas fezes de um felino silvestre na região Sul do Brasil. Os parasitos foram obtidos a partir de amostras fecais, sendo utilizadas quatro técnicas para os exames parasitológicos: exame direto, centrífugo-flutuação com sulfato de zinco (Técnica de Faust), sedimentação simples (Técnica de Hoffman) e Baermann-Moraes. Os parasitos foram identificados através de micrometria e morfologia, sendo esses: Ancylostoma sp., Toxocara sp., Trichuridae, Aelurostrongylus abstrusus, Alaria sp. e Spirometra sp. Estudos da fauna parasitária de animais silvestres são relevantes, tanto para o equilíbrio e saúde desses animais, como para o controle e prevenção de doenças transmitidas ao homem. Ancylostoma spp. foi identificado pela primeira vez em L. colocolo.

Molecular detection of Trypanosoma evansi (Kinetoplastida: Trypanosomatidae) in procyonids (Carnivora: Procyonidae) in Eastern Amazon, Brazil / Detecção molecular de Trypanosoma evansi (Kinetoplastida: Trypanosomatidae) em procionídeos (Carnivora: Procyonidae) na Amazônia Oriental, Brasil

ABSTRACT: The present study aimed to diagnose the natural infection of captive and free-living procyonids with Trypanosoma evansi in the states of Amapá and Pará, Brazil. From February 2012 to August 2013, whole blood samples and blood smears were obtained from 45 free-living procyonids and from nine procyonids kept in captivity in wild life refuges and zoobotanical parks in the states of Amapá and Pará. Whole blood samples were collected and kept at -20ºC for the detection of T. evansi DNA by PCR using the RoTat 1.2 forward and RoTat 1.2 reverse primers. In addition, the blood smears were processed and examined for the presence of trypomastigote forms of T. evansi. T. evansi DNA was detected in 18.52% (10/54) of the procyonids, namely, in captive crab-eating raccoons and captive and free-living coatis in Pará State. No trypomastigote forms were observed in the blood smears. DNA from T. evansi was detected in P. cancrivorus and N. nasua in Pará State, being this the first such report in P. cancrivorus.

RESUMO: O objetivo do presente trabalho foi realizar o diagnóstico da infecção natural por Trypanosoma evansi em procionídeos de vida livre e de cativeiro dos estados do Amapá e Pará, Brasil. Durante o período de fevereiro de 2012 a agosto de 2013, amostras de sangue total e esfregaços sanguíneos foram obtidos de 45 procionídeos de vida livre e de nove mantidos em cativeiro em mantenedores e Parques Zoobotânicos dos estados do Amapá e Pará. As amostras de sangue total foram coletadas e mantidas a -20ºC para pesquisa de DNA de T. evansi pela PCR utilizando-se os iniciadores RoTat 1.2 forward e RoTat 1.2 reverse. Os esfregaços sanguíneos também foram processados e examinados para a pesquisa de formas tripomastigotas do agente. DNA de T. evansi foi detectado em 18,52% (10/54) dos procionídeos, ocorrendo em mãos-peladas de cativeiro e quatis de vida livre e de cativeiro no estado do Pará. Não foram observadas formas tripomastigotas nos esfregaços sanguíneos. DNA de T. evansi foi detectado em P. cancrivorus e N. nasua no estado do Pará, sendo este o primeiro relato em P. cancrivorus.

Iron metabolism and its relationship to anemia and immune system in Trypanosoma evansi infected rats.

The aim of this study was to evaluate biochemical parameters of iron metabolism in rats experimentally infected with Trypanosoma evansi. To this end, 20 rats (Wistar) were intraperitoneally inoculated with blood containing trypomastigotes 10(6) (Group T) and 12 animals were used as negative control (Group C) and received saline (0.2 mL) through same route. Blood samples were collected by cardiac puncture on day 5 (C5, T5) and 30 (C30, T30) post-inoculation (pi) to perform complete blood count and determination of serum iron, transferrin, ferritin, total and latent iron fixation capacity, transferrin saturation and prohepcidin concentration. Also, bone marrow samples were collected, to perform Pearls staining reaction. Levels of iron, total and latent iron binding capacity and prohepcidin concentration were lower (P<0.05) in infected rats (T5 and T30 groups) compared to controls. On the other hand, levels of transferrin and ferritin were higher when compared to controls (P<0.05). The transferrin saturation increased on day 5 pi, but decreased on day 30 pi. The Pearls reaction showed a higher accumulation of iron in the bone marrow of infected animals in day 5 pi (P<0.01). Infection with T. evansi in rats caused anemia and changes in iron metabolism associated to the peaks of parasitemia. These results suggest that changes in iron metabolism may be related to the host immune response to infection and anemic status of infected animals.

Rangelia vitalii: changes in the enzymes ALT, CK and AST during the acute phase of experimental infection in dogs / Rangelia vitalii: mudanças nas enzimas ALT, CK e AST na fase aguda da infecção experimental em cães

Rangelia vitalii is a protozoon that causes diseases in dogs, and anemia is the most common laboratory finding. However, few studies on the biochemical changes in dogs infected with this protozoon exist. Thus, this study aimed to investigate the biochemical changes in dogs experimentally infected with R. vitalii, during the acute phase of the infection. For this study, 12 female dogs (aged 6-12 months and weighing between 4 and 7 kg) were used, divided in two groups. Group A was composed of healthy dogs (n = 5); and group B consisted of infected animals (n = 7). Blood samples were collected on days 0, 10, 20 and 30 after infection, using tubes without anticoagulant to obtain serum and analyze the biochemical parameters. An increase in alanine aminotransferase (ALT) on day 20 (P < 0.05) was observed. Also, increased creatine kinase (CK) and aspartate aminotransferase (AST) levels were observed throughout the experimental period (P < 0.05). No changes in the serum gamma-glutamyltransferase, urea and creatinine levels were observed. Thus, is possible to conclude that experimental infection with R. vitalii in dogs causes changes to the biochemical profile, with increased ALT, AST and CK enzyme levels.

Rangelia vitalii é um protozoário que causa doença em cães, sendo a anemia o achado laboratorial mais frequente. No entanto, existem poucos estudos sobre as alterações bioquímicas em cães infectados com o protozoário. Assim, este estudo tem como objetivo investigar as alterações bioquímicas de cães experimentalmente infectados com R. vitalii na fase aguda da infecção. Para o estudo, foram utilizados 12 cães fêmeas (com idade entre 6 a 12 meses e peso entre 4 a 7 kg), divididos em dois grupos. O grupo A (n = 5) foi composto de animais saudáveis e o grupo B (n = 7) de animais infectados. Amostras de sangue foram coletadas nos dias zero, dez, vinte e trinta PI, utilizando tubos sem anticoagulante para obtenção de soro e análise dos parâmetros bioquímicos. Foi observado um aumento na alanino aminotransferase (ALT) no dia 20 PI (P < 0,05) e aumento na creatinoquinase (CK) e aspartato aminotransferase (AST) em todo o período experimental (P < 0,05). Não foram observadas alterações séricas na gama-glutamiltransferase, uréia e creatinina. Portanto, é possível concluir que a infecção experimental por R. vitalii causa alterações no perfil bioquímico, com aumento na ALT, CK e AST.

Trypanocidal activity of human plasma on Trypanosoma evansi in mice / Atividade tripanocida do plasma humano sobre Trypanosoma evansi em camundongos

This study aimed to test an alternative protocol with human plasma to control Trypanosoma evansi infection in mice. Plasma from an apparently 27-year-old healthy male, blood type A+, was used in the study. A concentration of 100 mg.dL-1 apolipoprotein L1 (APOL1) was detected in the plasma. Forty mice were divided into four groups with 10 animals each. Group A comprised uninfected animals. Mice from groups B, C and D were inoculated with a T. evansi isolate. Group B was used as a positive control. At three days post-infection (DPI), the mice were administered intraperitoneally with human plasma. A single dose of 0.2 mL plasma was given to those in group C. The mice from group D were administered five doses of 0.2 mL plasma with a 24 hours interval between the doses. Group B showed high increasing parasitemia that led to their death within 5 DPI. Both treatments eliminated parasites from the blood and increased the longevity of animals. An efficacy of 50 (group C) and 80% (group D) of human plasma trypanocidal activity was found using PCR. This therapeutic success was likely achieved in the group D due to their higher levels of APOL1 compared with group C.

Este estudo teve como objetivo testar um protocolo alternativo com plasma humano para controlar a infecção por Trypanosoma evansi em camundongos. O plasma foi oriundo de um homem aparentemente saudável, com idade entre 27 anos e tipo de sangue A+. Foi detectada uma concentração de 100 mg.dL -1 de apolipoproteína L1 (APOL1) no plasma. Quarenta camundongos foram divididos em quatro grupos, contendo dez animais cada. Grupo A, composto de animais não infectados. Os roedores dos grupos B, C e D foram inoculados intraperitonealmente com um isolado de T. evansi. O Grupo B foi usado como um controle positivo. Três dias pós-infecção (DPI), os camundongos foram tratados com plasma humano. Uma dose única de 0,2 mL de plasma foi administrada nos roedores do grupo C. Os ratos do grupo D receberam cinco doses de 0,2 mL de plasma em intervalos de 24 horas. Os ratos do grupo B apresentaram parasitemia crescente, o que ocasionou a morte dos animais em 5 DPI. Ambos os tratamentos foram capazes de eliminar o parasito do sangue e aumentar a longevidade dos animais. O método da PCR detectou uma eficácia de 50% (grupo C) e 80% (grupo D) no tratamento com plasma humano. Este sucesso terapêutico obtido nos animais do grupo D provavelmente foi por receber maiores níveis de APOL1, comparado ao grupo C.

Gastrointestinal parasites of cavy (Cavia aperea aperea) in southern Brazil.

The aim of this study was to evaluate the gastrointestinal parasitism in Cavia aperea aperea (cavy), captured in the central region of Rio Grande do Sul State. Fecal samples from five free-living cavies were collected for research of parasites. Samples were analyzed by the centrifugal-flotation method with zinc sulfate and parasites were identified microscopically based on (oo)cyst and egg size and morphology. Cysts of Giardia sp. and (oo)cysts of Cryptosporidium sp. and Cystoisospora sp. were observed in one or more cavies. Eggs of Paraspidodera uncinata were observed in three of the five rodents. All infected animals showed mild infection by parasite. This is the first report of Giardia sp., Cryptosporidium sp. and Cystoisospora sp. in Cavia a. aperea.

Suscetibilidade de Trypanosoma evansi à anfotericina B / Trypanosoma evansi susceptibility to amphotericin B

O objetivo deste estudo foi avaliar a suscetibilidade do Trypanosoma evansi in vitro e in vivo à anfotericina B. Nos testes in vitro, foram utilizadas quatro concentrações (0,06; 0,25; 1,0; 4,0µg mL-1) de anfotecicina B frente a uma suspensão de T. evansi em solução tampão fosfato rico em glicose (PBS - glicose). Para avaliar a eficácia in vivo, foram utilizados 15 ratos parasitados com T. evansi. Em dois grupos de cinco ratos infectados, doses únicas diárias de 1 (grupo A) e de 3mg kg-1 (grupo B) foram administradas via intraperitonial durante 10 dias, e a parasitemia foi avaliada por meio de esfregaço sanguíneo. Grupo C (n=5) foi utilizado como grupo controle positivo, infectados com T. evansi e não tratados, e o grupo D (n=5), como controle negativo. Os ensaios in vitro evidenciaram suscetibilidade de 100 por cento do T. evansi à anfotericina B após 7h, em todas as concentrações avaliadas. Nos ratos, nem a maior dose testada curou os roedores, apesar de ter prolongado a vida destes em comparação à vida dos animais infectados, mas não tratados. Foi também investigada a função hepática e renal dos ratos após a terapia, e os parâmetros bioquímicos analisados mantiveram-se dentro da normalidade. Conclui-se que o T. evansi in vitro é suscetível à anfotericina B. A dose 3mg kg-1 testada aumentou a expectativa de vida de ratos infectados, porém não teve efeito curativo.

The aim of this study was to evaluate the Trypanosoma evansi susceptibility to amphotericin B in vitro and in vivo. Four concentrations (0.06, 0.25, 1.0, and 4.0µg mL-1) of amphotericin B were tested against a suspension containing T. evansi and phosphate buffer solution with glucose in the in vitro assay. Fifteen rats infected with T. evansi were used for the in vivo assay. Groups A (n=5) and B (n=5) received daily doses of 1 and 3mg kg-1 during 10 days and the parasitemia was estimated daily by microscopic examination of smears. The rats from group C (n=5) were the positive control and were infected but not treated. Rats from group D (n=5) were used as negative control. in vitro assays showed a 100 percent of susceptibility of T. evansi to amphotericin B after 7 hours, at all concentrations tested. The higher dose tested did not cure the rats, although treated rats had a longer life span in comparison to the non-treated group. Adverse effects on renal and hepatic hemodynamics were also researched. Biochemical parameters obtained were within the normal ranges. It was concluded that T. evansi is susceptible to amphotericin B in vitro. The dose of 3 mg kg-1 tested in rats increased life span, but did not cure the animals.

Primeiro registro de Trypanosoma vivax em bovinos no Estado do Rio Grande do Sul, Brasil / First report of Trypanosoma vixax in bovines in the State of Rio Grande do Sul, Brazil

Este estudo teve o objetivo de relatar a ocorrência de Trypanosoma vivax no Sul do Brasil. O protozoário foi diagnosticado em esfregaço sanguíneo de um bovino e a identificação baseada na morfologia das formas tripomastigotas e confirmada pela técnica de reação em cadeia de polimerases (PCR). O animal infectado apresentou sintomatologia compatível com a forma nervosa da infecção por T. vivax. Outros bovinos que compartilhavam o mesmo ambiente apresentaram resultados negativos para T. vivax por PCR. Este é o primeiro registro de T. vivax no Estado do Rio Grande do Sul e na região Sul do Brasil.

This study aimed at reporting the occurrence of Trypanosoma vivax in southern Brazil. The protozoan was diagnosed through peripheral blood smear evaluation of a bovine and confirmed by the evaluation of the trypomastigote forms morphology and by the Polimerase Chain Reaction (PCR). The animal showed clinical signs similar to the nervous form of the infection by T. vivax. Negative results for T. vivax were found in other bovines grazing in the same paddock. This is the first report of T. vivax in the state of Rio Grande do Sul and in the southern region of Brazil.

Patogenicidade de um isolado de Trypanosoma evansi em ratos inoculados com o parasito em sangue in natura e criopreservado / Trypanosoma evansi pathogenicity strain in rats inoculated with parasite in fresh and cryopreserved blood

O objetivo deste estudo foi investigar a patogenicidade do isolado de Trypanosoma evansi (LPV-2005), em ratos (Rattus norvergicus), sob influência da imunidade passiva, de diferentes concentrações e de meios de conservação. Para tanto, foram utilizados 36 Rattus norvergicus, fêmeas, separados em seis grupos homogêneos. Os roedores dos grupos A e B foram infectados com 10(5) T. evansi, e os animais dos grupos C e D foram infectados com 10(6) tripomastigotas/animal. Os grupos E e F foram utilizados como grupo controle negativo, isto é, inoculados com sangue in natura e criopreservado sem o parasito, respectivamente. O grupo A foi formado por ratos filhos de fêmeas infectadas com protozoário, mas curadas após tratamento. Os grupos B, C e D continham roedores que nunca tiveram contato com o isolado LPV-2005. Os grupos B e C diferiram quanto à dose inoculada do flagelado mantida em cultura viva (ratos Wistar). Já os ratos do grupo D foram infectados com sangue criopreservado em nitrogênio líquido. A patogenicidade do isolado foi avaliada a partir do período pré-patente, da evolução da parasitemia e da longevidade dos animais. O grupo D apresentou um período pré-patente superior aos demais grupos. Em relação à longevidade dos animais de cada grupo, foi verificada diferença estatística significativa (P<0,05). O grupo D apresentou um período de vida de 27,8 dias, e o grupo C, de apenas 4,8 dias. Os ratos de ambos os grupos controle mantiveram-se vivos por 50 dias, quando foram eutanasiados. Portanto, a preservação do inóculo testado e a dose infectante de T. evansi influenciam a patogenicidade do isolado LPV-2005 para ratos. A presença de anticorpos maternos em ratos não impede a infecção e mortalidade por T. evansi.

This study aimed to evaluate the Trypanosoma evansi strain pathogenicity (LPV-2005) in rats under passive immunity influence, of different concentrations and media preservation. Thirty six adult female Rattus norvergicus were separated in six equal groups. Groups A and B were inoculated with 10(5) T. evansi and groups C and D with 10(6) blood trypomastigotes per animal. Groups E and F were used as negative control in which the animals were inoculated with fresh and cryopreserved blood, without the parasite. Group A were composed of T. evansi infected born rats and cured females. Groups B, C and D were composed with animals never exposed to the LPV-2005 strain. All groups B and C animals received different doses of blood trypomastigotes kept in Wistar rats, while animals from group D were infected with cryopreserved blood kept in liquid nitrogen. The the strain pathogenicity was estimated by prepatency evaluation period, levels of parasitemia and animals longevity. Group D showed a longer prepatency period in comparison with other groups. The longevity of group D (27.8 days) was significantly different (P<0.05) from group C (4.8 days). Rats of the control group were euthanized 50 days postinfection. In conclusion, the tested inoculum-preservation methods and the infective dose of T. evansi influenced the pathogenicity of the LPV-2005 strain in rats. The presence of maternal antibodies did not prevent the infection and mortality of the rats by T. evansi.

Leptospirose e tritricomonose: isolamento em propriedade com problemas reprodutivos no sul do Brasil / Leptospirosis and tritrichomonosis: property in isolation with reproductive problems in southern Brazil / Leptospirosis y tritricomonosis: aislamiento en propiedad con problemas reproductivos en el sur de Brasil

O objetivo do estudo foi pesquisar quais os agentes etiológicos presentes em touros de duas propriedades rurais com elevados índices de repetição de cio. Da propriedade A foram analisados 23 touros, dos 58 integrados no rebanho, e na propriedade B foram analisados 15 reprodutores. Destes animais, foi colhido sangue para exame de leptospirose e realizado lavado prepucial para cultura de tritricomonose. Na propriedade A, em nenhuma das amostras avaliadas isolou-se trofozoítas de Tritrichomonas foetus. No soro de 15 touros (65,2%) testados para Leptospira foi verificada reação para um ou mais sorovares de L. hardjo, L. wolffi, L. icterohaemorrhagiae e L. butembo, em titulações que oscilaram entre 1/100 e 1/400. Na propriedade B não foi verificada titulação para leptospirose, mas em um dos animais isolaram e reproduziram trofozoìtas de T. foetus em laboratório. Do restante dos animais, cinco foram considerados suspeitos em microscópico óptico, não sendo confirmado o parasitismo em lâmina corada com Giemsa. Todos os bovinos da propriedade A foram submetidos ao tratamento com sulfato de estreptomicina em dose única de 25mg kg-1. O animal positivo para T. foetus foi descartado e os demais estão sendo monitorados mensalmente. Tanto Leptospirose como tritricomonose são enfermidades capazes de acarretar problemas reprodutivos em bovinos no sul do Brasil.

The aim of this study was to research the etiological agents present in bulls from a property with high estrus repetition rate. Twenty three out of 58 bulls in the property A and 15 of the property B were analyzed. Blood samples and preputial washes for leptospirosis and tritrichomonosis research were collected, respectively. In the property A, none of the evaluated animals showed the presence of Tritrichomonas foetus trophozoites, although 15 bulls (65,2%) tested for Leptospira reacted with one or more sorovars of L. hardjo, L. wolffi, L. icterohaemorrhagiae and L. butembo with titles between 100 and 400. Animals from property B were negative for leptospirosis and the isolation and reproduction of T. foetus in laboratory was possible only in one animal. Five other animals were considered suspect in the direct smear technique, although they were negative in the Giemsa staining method. All animals from property A received a single dose of streptomycin sulfate (25mg kg-1) which drastically reduced the estrus repetition rate. The animal with tritrichomonosis was taken out of the herd and the others have been monitored monthly. Even leptospirosis as tritrichomonosis are diseases directly linked with reproductive problems in southern Brazil.

El objetivo de este estudio fue investigar cuales son los agentes etiológicos presentes en toros de dos propiedades rurales con elevados índices repetitivos de celo. De la propiedad “A” fueron analizados 23 toros, de los 58 que integran el rebaño, en la propiedad “B” fueron analizados 15 reproductores. De estos animales, se extrajo muestras de sangre para examen de leptospirosis y se realizó lavado prepucial para cultivo de tritricomonosis. En la propiedad “A”, en ninguna de las muestras evaluadas se aisló trofozoítos de Tritrichomonas foetus, ya en el suero de 15 toros (65,2%) testeados para Leptospira fue verificada reacción para uno o más sorovares de L. hardjo, L. wolffi, L. icterohaemorrhagiae y L. butembo en titulaciones que oscilaron entre 1/100 y 1/400. En la propiedad “B” no fue verificada titulación para leptospirosis, pero en uno de los animales se aisló y se reprodujeron trofozoítos de T. foetus en laboratorio. En los demás animales, cinco fueron considerados sospechosos en microscopio óptico, no siendo confirmado el parasitismo en lámina coloreada con Giemsa. Todos los bovinos de la propiedad A fueron sometidos al tratamiento con sulfato de estreptomicina en dosis única de 25mg kg-1. El animal positivo para T. foetus fue descartado y los demás están siendo monitoreados mensualmente. Tanto Leptospirosis como tritricomonosis son enfermedades capaces de provocar problemas reproductivos en bovinos en el sur de Brasil.

Occurrence of Amblyomma longirostre in Ramphastos dicolorus in Southern Brazil / Ocorrência de Amblyomma longirostre em Ramphastos dicolorus no Sul do Brasil

The species Amblyomma longirostre Koch, 1844 is poorly known in the state of Rio Grande do Sul. Usually the adult stage could be found on Sphigurus spp. and the immatures on birds (Passeriformes). Although A. longirostre is distributed in the Neotropical region, from Panama to Uruguay, it also occurs in Central America and the United States. The aim of this study was to report that Ramphastos dicolorus Linnaeus 1766 is a new host record for this tick species.

A espécie Amblyomma longirostre Koch, 1844 é pouco conhecida no Estado do Rio Grande do Sul. Normalmente a fase adulta pode ser encontrada em porquinhos Sphigurus spp., e as fases imaturas podem ser encontradas em aves (Passeriformes). A. longirostre distribui-se na região neotropical, do Panamá ao Uruguai. Ela também ocorre na América Central e nos Estados Unidos. Este estudo tem como objetivo relatar que Ramphastos dicolorus Linnaeus 1766 é um novo hospedeiro dessa espécie de carrapato.

Trypanosoma evansi: hematologic changes in experimentally infected cats.

This study aimed at evaluating hemogram and erythropoietic changes in cats experimentally infected with Trypanosoma evansi. Thirteen adult female non-breeding Felix catus were separated into two groups: seven animals were infected with 10(8) trypomastigotes each, and six animals were used as negative controls. Animals were kept in air-conditioned rooms and blood smears were performed daily for 49 days. Blood samples were collected from the jugular vein at days 0, 7, 21, 35 and 49 and stored in blood-collecting tubes containing anticoagulant. Bone marrow was collected from the proximal epiphysis of the right femur at days 14 and 42 post-inoculation (PI). Total erythrocyte count, hematocrit and hemoglobin showed statistical differences among groups from the seventh day PI onwards (P<0.05). The mean corpuscular volume and the mean corpuscular hemoglobin concentration remained normal, characterizing a normocytic-normochromic anemia. Reticulocyte count increased in the infected group from the 21st day onwards, but remained near normal values suggesting a mild regenerative anemia. Moreover, the myeloid:erythroid ratio significantly reduced at day 42 PI, evidencing a bone marrow hematopoietic response. Based on these results we conclude that cats infected with T. evansi have normocytic, normochromic, regenerative anemia.