New artificial hematophagy system with attractive polymeric biofilm for maintenance of Culex quinquefasciatus (Diptera: Culicidae) in the laboratory.

BACKGROUND: Maintaining mosquito colonies in the laboratory requires a blood supply so that females' oocytes can mature and oviposition can take place. In this study, a new artificial hematophagy system for colonization and maintenance of Culex quinquefasciatus in the laboratory was developed and tested. METHODS: We developed an attractive polymeric biofilm including 25% L-lactic acid for use as a membrane in an artificial hematophagy system and compared the feeding rate of females with Parafilm-M®. We also evaluated the oviposition rate, larval survival and adult emergence of females fed through the attractive biofilm. RESULTS: The average percentage of female Cx. quinquefasciatus fed through the attractive biofilm was 87%, while only 20% became engorged with Parafilm-M® (p < 0.0001). Feeding through the attractive biofilm developed in this study produced high levels of evaluated biological parameters; the percentage of egg laying by females that underwent artificial hematophagy through the biofilm was 90%, with an average of 158 eggs per raft. From these eggs, 97% of the larvae hatched, of which 95% reached the pupal stage. The adult emergence rate corresponded to 93% of pupae. CONCLUSIONS: Insects fed with attractant through the biofilm system had a higher engorgement rate compared to those fed through Parafilm-M®. Our study is preliminary and suggests that polymeric biofilm has great potential for artificially feeding mosquitoes in the laboratory. Based on this research, new studies will be carried out with biofilm and different systems.

Gallium maltolate, a promising low toxicity drug with curative effect on mice chronically infected with Trypanosoma evansi.

Trypanosoma evansi is a flagellate protozoan that infects a wide range of hosts, especially horses. Clinically, the infection is characterized by rapid weight loss, anemia and mobility disorders. This study evaluated the efficacy of treatment gallium maltolate (GaM) in rats infected with T. evansi in the acute and chronic phases of the disease and its influence on the enzyme and blood parameters. 48 animals (Rattus norvegicus) were divided into 8 groups (A-H) of 6 animals each, namely: A: (negative control) uninfected; B: acutely infected positive control; C: chronically infected positive control; D: acutely infected, treated with GaM for 7 days post infection (p.i.); E: acutely infected treated with GaM for 3 days before infection (b.i) and 7 days p.i.; F: chronically infected, treated with GaM for 7 days p.i.; G: chronically infected, treated with GaM for 3 days b.i. and 7 days p.i.; and H: uninfected treated with GaM for 10 days. Acute infected animals (B, D and E) had a progressive increase in parasitemia and were died or euthanized before completing treatment days (5th days p.i.) as they had high parasitemia (over 100 field trypanosomes in the blood smear). Thus, it can be concluded that GaM was not effective against an acute infection. In untreated chronically infected animals (C) the parasitemia also increased progressively and they were euthanized on the 7th day p.i.. The chronically infected and treated animals (F and G) showed low parasitemia and after treatment became negative, showing no trypanosomes in the bloodstream until the 50th day of the experiment. Thus, we conclude that GaM was effective against chronic infections. In uninfected and treated animals (H) hematological, biochemical and enzymatic parameters had no significant changes when compared to the negative control group (A) demonstrating the low toxicity of GaM.

First reported case of Trichomonas gallinae infection in red-breasted toucan (Ramphastos dicolorus).

This study reports the infection and diagnosis of the protozoan morphologic complex Trichomonas gallinae in a baby red-breasted toucan (Ramphastos dicolorus). Nodular lesions on the soft palate and edema in the oral cavity were observed macroscopically. Microscopically, a granuloma with multiple layers of necrosis interspersed with inflammatory polymorphonuclear infiltrates was observed. Parasitism was confirmed by parasitological diagnosis, isolation of the flagellates in culture medium, and Polymerase Chain Reaction (PCR) using 5.8S ribosomal RNA (rRNA). Flanking internal transcribed spacer (ITS) gene regions were amplified by polymerase chain reaction, and the sequences were analyzed phylogenetically using MEGA 11 software. Phylogenetic analysis based on ITS1/5.8S rRNA/ITS2 sequences demonstrated high nucleotide identity with two Trichomonas sequences available in GenBank, which were more closely related to T. vaginalis (99%) than to T. gallinae (98%). In addition to being potential transmitters of this protozoan, rigorous monitoring of infectious and parasitic diseases in wild bird populations is essential for their preservation. The forms of transmission of Trichomonas sp. favor the occurrence of the disease in many non-Columbiformes species, which is essential for the monitoring of this disease in wild birds.

Activity of compounds derived from benzofuroxan in Trichomonasvaginalis.

Trichomoniasis is a sexually transmitted infection caused by the protozoan Trichomonas vaginalis. Currently, trichomoniasis is treated with the class of nitroimidazoles, namely, metronidazole; however, resistant isolates and strains have been reported. The compounds derived from benzofuroxan are biologically active heterocycles. This study evaluated the in vitro antiparasitic activity of these compounds in trophozoites of T. vaginalis and determined the mean inhibitory concentration (IC50), minimum inhibitory concentration (MIC), mortality curve, and cytotoxicity. The compounds were named EH1, EH2, EH3, and EA2 and tested in various concentrations: 100 to 15 µM (EH1 and EH2); 100 to 5 µM (EH3); and 100 to 25 µM (EA2), respectively. The greatest efficacy was observed in the highest concentrations in 24 h, with inhibition of approximately 100% of trophozoites. Compounds EH2 and EH3 had the lowest MIC: EH2 (35 µM) and EH3 (45 µM), with IC50 of 11.33 µM and 6.83 µM, respectively. Compound EA2 was effective at the highest concentrations. The activity of the compounds in T. vaginalis started in the first hour of incubation with 90% inhibition; after 12 h, inhibition >95% was observed. Compound EH1 showed the lowest activity, with the highest activity between 12 and 24 h after incubation. These results demonstrate that benzofuroxan derivatives are promising compounds for the in vitro treatment of T. vaginalis.

Microscopic alterations in Fasciola hepatica treated with the essential oils of Pelargonium graveolens and Citrus aurantium.

Fasciolosis is one of the most important parasitic diseases in animal health, affecting mainly ruminants, causing economic and productivity losses. This study aimed to evaluate the in vitro ovicidal and adulticidal activity of essential oils (EOs) from Pelargonium graveolens (geranium) and Citrus aurantium (sour orange) on Fasciola hepatica. Performed Gas Chromatography of EOs P. graveolens and C. aurantium, with major compound, citronellol (31.37 %) and limonene (93.89 %), respectively. For the cytotoxicity assay, the sour orange EO showed to be promising when used in lower concentrations. For the ovicidal tests, the eggs were incubated with geranium EOs at concentrations from 4.5 to 0.03375 mg/mL and sour orange at concentrations from 4.25 to 0.031875 mg/mL, along with controls. The viable eggs were counted on the 14th day post-incubation. Adult forms of F. hepatica were incubated containing the EOs and observed for 24 h after treatment, as well as the control groups. Later the specimens were fixed for histological analysis. Geranium and sour orange EOs in trematode eggs at the concentrations tested were 100 % effective in inactivating hatching (p < 0.05) when compared to the untreated control. In the adulticidal test, the essential oil of P. graveolens at both concentrations tested (0.0675 and 0.03375) within 15 h, promoted the death of flukes. For C. aurantium, 18 h was enough to inactivate all specimens, up to a concentration of 0.06375. The histological analysis, observed the accumulation of liquid in the tegument in the specimens incubated in C. aurantium and P. graveolens, with vacuolization in the tegument and spines, preventing externalization. The results of the study present OEs with efficient ovicidal and adulticidal activity.

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.

Diminazene aceturate liposomes: morphometric and biochemical liver, kidney, and spleen of rats infected with Trypanosoma evansi.

The aim of this study was to evaluate the effect of treatment with liposomal (L-DMZ) and conventional (C-DMZ) diminazene aceturate formulations on hepatic and renal functions of rats, experimentally infected with Trypanosoma evansi. For this purpose, 72 Wistar rats (Rattus norvegicus) were divided into six groups (A, B, C, D, E, and F). Each group was subdivided into two other subgroups in order to assess the biochemical and histological results on days 7 and 40 post-treatment (PT). Treatments were carried out based on two different therapeutic protocols: L-DMZ and C-DMZ at 3.5mg/kg(-1), single dose (groups C and D), and five successive doses within intervals of 24h (groups E and F). Groups A and B corresponded to uninfected and infected (without treatment) animals, respectively. Sample collections were held on days 7 and 40 PT for the assessment of hepatic [alkaline phosphatase (AP), alanine transferase (ALT), albumin, gamma glutamil transferase (GGT) and renal functions (creatinine and urea). Additionally, the histology of fragments of liver, kidney, and spleen was performed. Animals in group B showed a significant increase in AP, GGT, ALT, and urea when compared with group A. On day 7 post-inoculation (PI), the biochemical analysis showed a reduction (P<0.05) of AP and GGT, while the levels of urea were increased in groups C, D, E, F. On day 40 PT, ALT was increased in these same groups when compared with group A. In histopathology, changes in liver samples were observed on day 7 PT in groups D and F, especially regarding the area and density of the hepatocytes. Renal analysis exhibited changes in glomerular space, glomerular, and corpuscular areas in group E. Therefore, these results allowed us to conclude that the treatment with L-DMZ and C-DMZ led to variable biochemical changes, which defined the functions of the liver and kidneys of treated animals, since the main histopathology alterations were observed in animals treated with liposomes, at their higher dosages. Thus, treatments with L-DMZ and C-DMZ in five consecutive doses were effective although being followed by liver toxicity.

Liposomes produced by reverse phase evaporation: in vitro and in vivo efficacy of diminazene aceturate against Trypanosoma evansi.

This study aimed to develop and test the in vitro and in vivo effectiveness of diminazene aceturate encapsulated into liposomes (L-DMZ) on Trypanosoma evansi. To validate the in vitro tests with L-DMZ, the efficacy of a commercial formulation of diminazene aceturate (C-DMZ) was also assessed. The tests were carried out in culture medium for T. evansi, at concentrations of 0.25, 0.5, 1, 2 and 3 µg mL(-1) of L-DMZ and C-DMZ. A dose-dependent effect was observed for both formulations (L-DMZ and C-DMZ), with the highest dose-dependent mortality of trypomastigotes being observed at 1 and 3 h after the onset of tests with L-DMZ. The results of in vivo tests showed the same effects in the animals treated with L-DMZ and C-DMZ in single doses of 3.5 mg kg(-1) and for 5 consecutive days (3.5 mg kg(-1) day(-1)). It was possible to conclude that T. evansi showed greater in vitro susceptibility to L-DMZ when compared with C-DMZ. In vivo tests suggest that treatment with the L-DMZ and C-DMZ showed similar efficacy in vivo. The potential of the formulation developed in this study was clearly demonstrated, as it increased the efficacy of the treatment against trypanosomosis, but more studies are needed to increase the effectiveness in vivo.

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.

Ocorrência de Giardia sp. e Cryptosporidium sp. em Leopardus weidii de vida livre / Occurrence of Giardia sp. and Cryptosporidium sp. in Leopardus weidii of wild life

Leopardus weiidi (gato-maracajá) é um mamífero neotropical de ampla distribuição no Brasil. No município de Cachoeira do Sul, RS, um exemplar foi capturado para tratamento, quando se coletou uma amostra de fezes, que foi analisada pelo método de centrífugo-flutuação com sulfato de zinco para pesquisa de parasitos. Foi observada elevada infecção por cistos de Giardia sp. e oocistos de Cryptosporidium sp.. Com base nos resultados, conclui-se que L. weeidi é hospedeiro desses protozoários.

Leopardus weiidi (gato-maracajá) is a neo-tropical mammal with has a wide distribution in Brazil. In the municipality of Cachoeira do Sul, RS, a fecal sample from a specimen captured for treatment was collected and analyzed by the centrifugal-flotation method with zinc sulfate for research of parasites. A large number of cysts of Giardia sp. and oocysts of Cryptosporidium sp. was observed. The results indicate that L. weeidi is a host of these protozoa.

Parasitismo por Giardia sp. e Cryptosporidium sp. em Coendou villosus / Parasitism by Giardia sp. and Cryptosporidium sp. in Coendou villosus

O objetivo deste trabalho foi verificar o possível parasitismo por Giardia sp. e Cryptosporidium sp. em amostras de fezes de ouriço-cacheiro (Coendou villosus). As amostras foram analisadas pelo método de centrífugo-flutuação com sulfato de zinco e apresentaram elevada infecção por cistos de Giardia sp. e por oocistos de Cryptosporidium sp., embora os animais não apresentassem sinal clínico decorrente disso.

This research was aimed at verifing the possible parasitism by Giardia sp. and Cryptosporidium sp. in porcupine (Coendou villosus) faeces samples. Samples were analyzed by the centrifugal-flotation method with zinc sulphate and showed high infection by cysts of Giardia sp. and by oocysts of Cryptosporidium sp., although the animals did not show any associated clinical sign.

Infecção via oral por Trypanosoma evansi em animais de laboratório / Oral infection by Trypanosoma evansi in rats and mice

Testou-se a infecção de Trypanosoma evansi pela via oral em ratos e camundongos, através de sangue contaminado de ambas as espécies. Dez ratos e dez camundongos foram alocados em quatro grupos iguais A e B (ratos), C e D (camundongos). Os grupos A e C receberam sangue contaminado de um rato e o grupo B e D de um camundongo, através de uma sonda. O volume de sangue administrado foi de 0,2ml, o qual apresentava uma concentração de 10(7) tripanossomas ml-1. Os animais foram mantidos em temperatura e umidade constantes (25°C e 80 por cento UR), sendo realizados esfregaços sanguíneos diários para identificar o período pré-patente e a evolução do parasita na circulação. Nos grupos A e B, o período pré-patente variou de 19 a 25 dias, e o período entre a detecção dos parasitas e a morte dos animais foi em média de 12,7 dias. Os camundongos do grupo C e D não apresentaram infecção pelo parasita, sendo estes avaliados por 60 dias. Os ratos foram susceptíveis a infecção por T. evansi pela via oral; entretanto, os camundongos não se contaminaram com o protozoário por via digestiva.

In this research, Trypanosoma evansi infection was tested in rats and mice by oral ingestion of contaminated blood. Groups of ten rats and ten mice were disposed in four experimental groups: A and B (rats), C and D (mice). The groups A and C were contaminated by rat-contaminated blood; B and C groups by mouse-contaminated blood. The blood was given using a probe filled with 0.2ml of contaminated blood with 10(7) trypanosomes ml-1. These animals were maintained at constant temperature and humidity (25°C and 80 percent UR). Dairy blood smear were done to identify the prepatent period and evolution of parasite in the circulation. In the A and B groups, the pre latency period varied from 19 to 25 days and the period of parasite detection and animals death was an average of 12.7 days. The C and D groups did not present infection by the parasite even when evaluated for 60 days. In conclusion, the rats had oral infection by T. evansi but this protozoan couldnÆt contaminate the mice by digestive path.

Temnocefalídeo em tartaruga de água doce, Hydromedusa tectifera, da região central do Rio Grande do Sul / Temnocephalid in freshwater turtle, Hydromedusa tectifera, from central region of Rio Grande do Sul

O objetivo deste trabalho foi relatar a presença de Temnocephala sp. em tartaruga de água doce (Hydromedusa tectifera) na região central do Rio Grande do Sul, Brasil. Os platelmintos foram encontrados fixados no corpo, membros e casco do hospedeiro, sendo coletados através de raspagem cutânea. No laboratório, foram montados entre lâminas de vidro e identificados como pertencentes ao gênero Temnocephala.

This study was aimed at reporting the presence of Temnocephala sp. in fresh water turtle (Hydromedusa tectifera) central area of Rio Grande do Sul, Brazil. The platyhelminths have been found fastened in body, members, and shell of the host being collected through cutaneous scratching. In the laboratory they were assembled between glass sheets and identified as being belonging to the Temnocephala genus.

Ocorrência de Eimeria ichiloensis em capivaras (Hydrochaeris hydrochaeris) de criatório / Ocurrencia de Eimeria ichiloensis en capibaras (Hidrochoerus hidrochaeris) de creación / Occurrence of Eimeria ichiloensis in capybaras (Hydrochaeris hydrochaeris) bred in farming

O objetivo deste trabalho foi relatar o parasitismo por Eimeria ichiloensis em capivaras (Hydrochaeris hydrochaeris). Foram analisadas 10 amostras de fezes de capivaras, provenientes de um criatório legalizado, onde eram criadas em ambiente com área de brejo e açude no município de Arroio do Meio, Rio Grande do Sul, Brasil. As fezes foram analisadas através do método de centrífugo-flutuação com sulfato de zinco. Nas amostras, observou-se infecção leve por oocistos de E. ichiloensis, no entanto, os roedores não apresentaram sinais clínicos da enfermidade

This paper reports parasitism by Eimeria ichiloensis in capybaras (Hydrochaeris hydrochaeris). Ten fecal samples of animals from a legal nursery were analyzed. The animals were created in an environment with swamp and dam in the municipality of Arroio do Meio, Rio Grande do Sul state, Brazil. Feces were analyzed by the zinc sulphate centrifugalflotation method. Mild infection by oocytes of E. ichiloensis was observed in the samples, although rodents presented no clinical signs of the disease

El objetivo de esta investigación fue relatar el parasitismo por Eimeria ichiloensis en capibaras (Hydrochoerus hydrochaeris). Fueron analizadas 10 muestras de excrementos de capibaras, provenientes de una creación legalizada, donde eran creadas en pantanos y represa en el municipio de Arroio do Meio, Rio Grande do Sul, Brasil. Los excrementos fueron analizados a través del método centrífugo fluctuación con sulfato de zinc. En las muestras, se observó infección leve por oocistos de E. ichiloensis, sin embargo, los roedores no presentaron señales clínicos de la enfermedad