Ultrastructural characterization and quantification of hemocytes in engorged female Amblyomma sculptum ticks.

Amblyomma sculptum (formerly Amblyomma cajennense) ticks have been implicated in the transmission of pathogens that cause diseases in animals and humans. Their wide geographic distribution and high impact on animal health and zoonotic disease transmission highlight the importance of studying and implementing effective control measures to mitigate the risks associated with this tick species. The aim of this study was to quantify and characterize the morphology and the ultrastructure of different types of hemocytes in the hemolymph in engorged A. sculptum females fed on rabbits. The hemolymph samples were collected by perforation of the cuticle in the dorsal region. Hemocyte types, sizes, and differential counts were determined using light microscopy, while ultrastructural analysis of hemocytes was performed using transmission electron microscopy. The average number of total hemocytes in the hemolymph was 1024 ± 597.6 cells µL-1. Five morphologically distinct cell types were identified in A. sculptum females: prohemocytes (6 % ± 8.8), plasmatocytes (10 % ± 7.7), granulocytes (78 % ± 12.2), spherulocytes (5 % ± 4.48), and oenocytoids (1 % ± 1.6). In general, prohemocytes were the smallest hemocytes. The ultrastructural morphology of A. sculptum hemocytes described in the present study agrees with the findings for other hard ticks. This is the first study to investigate ultrastructural characteristics of hemocytes of female A. sculptum ticks.

Do combinations of fipronil, eugenol and carvacrol have synergistic effects against Rhipicephalus sanguineus?

The tick Rhipicephalus sanguineus is one of the main ectoparasites that affects dogs, causing direct and indirect damage to parasitized animals. Currently, infestation control is mainly carried out by using synthetic acaricidal drugs. However, a decrease in efficacy and an increase in resistance to the main therapeutic protocols against tick infestations have been increasingly reported and confirmed, a factor that has driven research into the potential acaricide activity of natural compounds, including in association with synthetic molecules. The aim of this work was to evaluate whether the combinations of fipronil (FIP) and eugenol (EUG), FIP and carvacrol (CAR), and EUG and CAR would have synergistic effects against immature and unfed adult stages of R. sanguineus through in vitro bioassays. Bioassays were carried out using the larval packet test (FAO 2004) adapted for nymphs and adults. The synergistic activity was explored by combining each solution, based on the estimated LC50, in a 1:1 ratio (FIP: EUG, FIP: CAR and EUG: CAR). CompuSyn software was used to evaluate the various pairwise combinations of FIP, EUG and CAR, checking if there was synergism or antagonism between them. FIP and EUG and FIP and CAR showed combination index (CIn) values above 1.45, indicating antagonism. The synergistic activity between EUG and CAR was verified against all unfed phases of R. sanguineus, since the CIn was below 0.70, a value that indicates synergism. The combination of fipronil with either eugenol or carvacrol presented antagonistic effects against R. sanguineus larvae. On the other hand, carvacrol and eugenol had excellent pharmacological synergism against all tick stages with mortality values in the range of 80 to 100%, including the adult stage, which is less susceptible than immature stages.

Prevalence of gastrointestinal parasites in domestic cats (Felis catus) diagnosed by different coproparasitological techniques in the municipality of Seropédica, Rio de Janeiro.

The objectives of this work were to investigate the occurrence of parasites in feces of cats, compare different coproparasitological techniques for their diagnosis and determine associations with parasitism. The samples were processed using three different coproparasitological techniques: centrifugal flotation in sucrose, centrifugal flotation in ZnSO4 and simple sedimentation. The parasitic association between parasitism and variables such as age, sex and fecal consistency was performed using the chi-square test or the G test with a significance level of 5%. A total of 237 samples were analyzed, of which 93 (39.2%) were positive, being Ancylostoma spp. (17.3%), Giardia intestinalis (12.2%), Platynosomum illiciens (8.0%), Cystoisospora spp. (6.3%), Toxoplasma gondii/ Hammondia hammondi (3.4%), Diphyllobothriidae (2.1%), Toxocara spp. (1.7%), Dipylidium caninum (1.3%) and Mesocestoides spp. (0.8%). In the parasitism association analysis, it was possible to verify a statistical difference in the age category for Cystoisospora spp. (p=0.001) observing a strong relationship between parasitism and young animals, the association with sex proved to be important for P. illiciens (p<0.001) with a higher frequency of parasitized females and fecal consistency revealed to be related to the parasites G. intestinalis (p=0.007) and P. illiciens (p=0.033) showing a higher number of positive animals for these parasites with normal fecal consistency. In conclusion, we observed a higher occurrence of Ancylostoma spp. and G. intestinalis in fecal samples from domestic cats received in routine diagnoses and the presence of other parasites with zoonotic potential, as well as the relationship of these diagnosed parasites with the categories sex, age and fecal consistency.

Evaluation of the susceptibility to fipronil of Rhipicephalus microplus larvae from egg masses incubated at different times of oviposition.

The objective of this work was to evaluate the susceptibility of R. microplus larvae from different oviposition times to fipronil. The LPT was performed in sextuplicate, at concentrations of 18.75, 37.5, 75, 150 and 300 µg.mL-1. The LC50 found for the egg masses incubated with +7, +14 and +21 days were respectively 105.87, 110.71 and 121.22 µg.mL-1. The larvae originating from egg masses from the same group of engorged females, incubated on different days, presented similar mortality rates compared to the evaluated fipronil concentrations, facilitating the maintenance of laboratory colonies of this tick species.

O objetivo deste trabalho foi avaliar a susceptibilidade de larvas de R. microplus oriundas de diferentes momentos da oviposição frente ao fripronil. O TPL foi realizado em sextuplicata, nas seguintes concentrações 18,75; 37,5; 75; 150; 300 µg.mL-1. Não houve diferença estatística entre as mortalidades das larvas oriundas de posturas incubadas nos dias mais sete, +14 e +21, expostas ao fipronil. As CL50 encontradas para as posturas incubadas com +7, +14 e +21 dias foram respectivamente 105,87; 110,71 e 121,22 µg.mL-1. Observou-se que as larvas oriundas de posturas, do mesmo grupo de fêmeas ingurgitadas, incubadas em dias diferentes apresentam taxas de mortalidade parecidas frente as concentrações de fipronil avaliadas, facilitando a manutenção das colônias laboratorias desta espécie de carrapato.

Updated spatial analysis of areas of occurrence of bovine cysticercosis in the state of Espírito Santo, Brazil.

The risk of bovine cysticercosis is mutable over time, since population habits, local infrastructure, and bovine herd size change through the years. Hence, the aim of this study was to update the risk map of bovine cysticercosis occurrence in Espírito Santo State, Brazil, by choosing the following risk factors: inadequate sewer service; bovine population per municipality; risk of flooding; and land use and occupation, examined with a mathematical model using the GIS program ArcGis 10.5. The risk map of occurrence demonstrated that the state is composed of 12.8, 24.5, 36.2 and 26.5% of areas of very high, high, medium, and low risk, respectively. The northern area stood out for areas of very high risk, with the cities of Linhares and São Mateus accounting for 99% of the area with this classification. Of the state's 78 municipalities, 15 had a portion of their territory with very high risk, 11 had over 50% of the territory with high risk and all except Irupi (100% of low risk) had some percentage of medium risk. These findings demonstrate the areas where actions to prevent bovine cysticercosis should be preferentially conducted.

O risco de cisticercose bovina é mutável ao longo do tempo, uma vez que os hábitos populacionais, a infraestrutura local e o tamanho do rebanho bovino mudam ao longo dos anos. Assim, o objetivo deste estudo foi atualizar o mapa de risco da ocorrência da cisticercose bovina no Estado do Espírito Santo, Brasil, escolhendo os seguintes fatores de risco: saneamento sanitário inadequado; população bovina por município; risco de inundação; e uso e ocupação do solo, examinados com um modelo matemático usando o programa SIG ArcGis 10.5. O mapa de risco de ocorrência demonstrou que o estado é composto por 12,8, 24,5, 36,2 e 26,5% de áreas de risco muito alto, alto, médio e baixo, respectivamente. A zona norte destacou-se como área de risco muito alto, com os municípios de Linhares e São Mateus respondendo por 99% da área com essa classificação. Dos 78 municípios do estado, 15 tinham uma parcela de seu território com risco muito alto, 11 tinham mais de 50% do território com risco alto e todos exceto Irupi (100% de risco baixo) tinham algum percentual de risco médio. Esses achados demonstram as áreas onde as ações de prevenção da cisticercose bovina devem ser preferencialmente realizadas.

Ocimum gratissimum essential oil and eugenol against Ctenocephalides felis felis and Rhipicephalus sanguineus: In vitro activity and residual efficacy of a eugenol-based spray formulation.

Fleas and ticks are among the main ectoparasites that affect pets. The indiscriminate and incorrect use of chemical antiparasitics may be related to increased insect resistance and environmental contamination, requiring prospection for active ingredients that are less harmful to animals, humans and the environment. The use of essential oils and their isolated compounds has been reported as a potential alternative to synthetic antiparasitics, but there is a lack of studies involving the design and development of stable and safe natural products-based formulations. Therefore, the aim of this study was to establish LC50 and LC90 of Ocimum gratissimum essential oil and eugenol on immature stages and adults of Ctenocephalides felis felis and Rhipicephalus sanguineus; and to design and to determine the in vitro efficacy and residual effect of a natural product-based spray formulation for flea and tick control in pets. Bioassays were carried out according to the filter paper impregnation technique for fleas and through the larval packet test for ticks. O. gratissimum essential oil and eugenol presented pulicidal and acaricidal activity in vitro against immature stages and adults of C. felis felis and immature stages of R. sanguineus. The greater potency of eugenol against fleas and ticks led to the choice of eugenol as the active ingredient in the pharmaceutical form developed. The developed eugenol-based sprays presented adequate physical and chemical characteristics and stability, had pulicidal and acaricidal efficacy after 24 h and residual effect against fleas for up to 48 days.

Platynosomum illiciens (Trematoda: Dicrocoeliidae) in a hybrid marmoset (Callithrix sp.) in the Municipality of Seropédica, RJ, Brazil - Case report.

Platynosomum illiciens is a liver trematode encountered infecting mainly felids although it has also been reported in birds and in additional mammalian species, including non-human primates. The current study reports a natural P. illiciens infection primate of the genus Callithrix. The diagnosis was made using a combination of copro-parasitological techniques, morphological evaluation of adult specimens recovered from the liver during necropsy, and molecular analyses. Eggs were brown in color, oval, operculated, and contained a miracidium. Adult specimens recovered during necropsy were measured and showed dimensions compatible with P. illiciens. Molecular characterization of the trematode involved amplification by polymerase chain reaction (PCR), in combination with nucleotide sequencing, of an approximately 900 base pairs fragment corresponding to 18S-ITS1-5.8S ribosomal DNA. Sequenced amplicons showed 100% nucleotide identity with sequences deposited in the GenBank database as derived from specimens of P. illiciens recovered from cats in Malaysia and Brazil. It was concluded that the morphological and molecular analyses presented herein, confirmed the identification of the trematode recovered as P. illiciens.

In vitro acaricidal activity of different ectoparasiticide classes against Amblyomma sculptum larvae.

Zoonoses are major causes of morbidity and mortality worldwide. Among them, Brazilian Spotted Fever (BSF) is an important one that occurs in some regions of South America and can be transmitted by the "star tick" Amblyomma sculptum. Application of acaricides against the larval stage is important as strategy of population control. However, there is still a deficiency of studies on chemical control of A. sculptum and the present work aims to evaluate the in vitro acaricidal activity of cypermethrin, flumethrin, deltamethrin, fipronil, coumaphos and chlorpyrifos against A. sculptum larvae. Bioassays were performed using the larval immersion test method. A discriminatory analysis between the antiparasitic classes most used for tick control was carried out, which made it possible to determine the classes with higher potential for controlling A. sculptum larvae. Our results showed that A. sculptum larvae present highest sensitivity to the synthetic pyrethroid group, followed by the phenylpyrazole, organophosphate and macrocyclic lactone groups. These findings may support studies on improvement of tick control as in animals as in the environment.

Development of Ctenocephalides felis felis (Siphonaptera: Pulicidae) in different substrates for maintenance under laboratory conditions.

The aim of this study was to evaluate the efficiency of different substrates for larval development of Ctenocephalides felis felis during its biological cycle. Eight hundred eggs of C. felis felis from a flea maintenance colony were used. Different diets were formulated, in which the main substrates were meat flour, powdered milk, sugar, lyophilized bovine blood, tick metabolites and lyophilized egg. The flea eggs were placed in test tubes (10 per tube) and approximately 2 g of the diet to be tested was added to each tube. There were 10 replicates for each substrate. After 28 days, each tube was evaluated individually for the presence of pupae and emerged adults. The following percentages of the larvae completed the cycle to the adult stage: 67% in diets containing tick metabolites; 55%, meat flour; 39%, dehydrated bovine blood; 14%, powdered milk; and less than 1% in diets containing sugar, lyophilized bovine blood, lyophilized egg or wheat bran. It was concluded that among the diets tested, the one constituted by tick metabolites as the substrate was shown to be the most satisfactory for maintaining a laboratory colony of C. felis felis, followed by the one containing meat flour.

Activity of Syzygium aromaticum essential oil and its main constituent eugenol in the inhibition of the development of Ctenocephalides felis felis and the control of adults.

The aim of this study was to evaluate the in vitro activity of Syzygium aromaticum essential oil (SAEO) and its main constituent eugenol (EG) against adult fleas and their action in the maturation of eggs into adults of Ctenocephalides felis felis. In order to evaluate the pulicidal activity, 10 adult fleas were exposed to a filter paper impregnated with SAEO and EG at increasing concentrations of 0.047; 0.094; 0.188; 0.375; 0.750; 1.50; 3.00; 6.00; 12.00 and 24.00 µg cm-2. Flea mortality was evaluated 24 and 48 h after exposure. In order to evaluate the inhibition of the maturation of eggs into adults, 10 eggs were exposed to filter paper impregnated with SAEO and EG at the same concentrations used in the pulicidal test, and the evaluation was performed 30 days after incubation. Untreated repetitions were maintained in both studies (control group). The lethal concentration 50 (LC50) of pulicidal activity to SAEO was 5.70 µg cm-2 in 24 h and 3.91 µg cm-2 in 48 h. The LC90 was 16.10 µg cm-2 and 15.80 µg cm-2 in 24 and 48 h, respectively. The LC50 of inhibition of the maturation of eggs into adults was 0.30 µg cm-2 and the LC90 3.44 µg cm-2. The LC50 of pulicidal activity to EG was 2.40 µg cm-2 in 24 h and 1.40 µg cm-2 in 48 h; the LC90 was 8.10 µg cm-2 and 3.70 µg cm-2 in 24 h and 48 h, respectively. The LC50 of inhibition of the maturation of eggs into adults was 0.10 µg cm-2 and the LC90 0.68 µg cm-2. Based on the results obtained, it is possible to conclude that the both SAEO and EG have in vitro pulicidal activity as well as acting as inhibitors of the maturation of eggs into adults of the flea C. felis felis.

Development of Ctenocephalides felis felis (Siphonaptera: Pulicidae) in different substrates for maintenance under laboratory conditions / Desenvolvimento de Ctenocephalides felis felis (Siphonaptera: Pulicidae) em diferentes substratos para manutenção em condições laboratoriais

Abstract The aim of this study was to evaluate the efficiency of different substrates for larval development of Ctenocephalides felis felis during its biological cycle. Eight hundred eggs of C. felis felis from a flea maintenance colony were used. Different diets were formulated, in which the main substrates were meat flour, powdered milk, sugar, lyophilized bovine blood, tick metabolites and lyophilized egg. The flea eggs were placed in test tubes (10 per tube) and approximately 2 g of the diet to be tested was added to each tube. There were 10 replicates for each substrate. After 28 days, each tube was evaluated individually for the presence of pupae and emerged adults. The following percentages of the larvae completed the cycle to the adult stage: 67% in diets containing tick metabolites; 55%, meat flour; 39%, dehydrated bovine blood; 14%, powdered milk; and less than 1% in diets containing sugar, lyophilized bovine blood, lyophilized egg or wheat bran. It was concluded that among the diets tested, the one constituted by tick metabolites as the substrate was shown to be the most satisfactory for maintaining a laboratory colony of C. felis felis, followed by the one containing meat flour.

Resumo Este trabalho teve como objetivo avaliar a eficiência de diferentes substratos no desenvolvimento larval de Ctenocephalides felis felis durante seu ciclo biológico. Foram utilizados 800 ovos de C. felis felis, oriundos de colônia de manutenção de pulgas. Diferentes dietas foram formuladas, contendo como substratos principais a farinha de carne, leite em pó, açúcar, sangue bovino liofilizado, metabólitos de carrapato e ovo liofilizado. Foram distribuídos 10 ovos por tubo de ensaio, aos quais foram acrescidos as dietas a serem testadas, realizando-se10 repetições para cada substrato. Após 28 dias, cada tubo foi avaliado individualmente pela presença de pupas e adultos emergidos. Nas dietas que continham metabólitos de carrapato, 67% das larvas completaram o ciclo até a fase adulta; 55% nas que continham farinha de carne; 39% contendo sangue bovino desidratado; 14% com leite em pó, e menos de 1% em dietas contendo açúcar, sangue bovino liofilizado, ovo liofilizado e farelo de trigo. Conclui-se que, entre as dietas testadas, a constituída por metabólitos de carrapato como substrato, mostrou-se a mais satisfatória para a manutenção de colônia laboratorial de C.felis felis, seguida da que continha farinha de carne.

Microscopic alterations in Fasciola hepatica from sheep treated with albendazole / Alterações microscópicas em Fasciola hepatica de ovelhas tratadas com albendazol

Abstract Currently, albendazole is one of the most commonly used drugs because of its affordability. The objective was to evaluate the histopathology of Fasciola hepatica specimens. For this, the efficacy test was performed on sheep treated with albendazole at the dose recommended for F. hepatica, in which the helminths recovered at necropsy were counted and separated for histology. Spermatogenic cells from parasites recovered from treated and control sheep were examined by microscopy. The fecal egg-count reduction test revealed 97.06% efficacy of albendazole in the treatment of F. hepatica. Changes in testicular tubule cells started 48 hours after treatment and became evident within 72 hours, at which point it became difficult to identify cell types. Primary and secondary spermatogonia became increasingly rare and intercellular vacuolization was more evident. Signs of apoptosis, with pycnotic nuclei and evidence of keriorrexia were observed at all times. Cell debris was identified 96 hours after treatment. The results indicated that parasitic spermatogenesis was severely affected by albendazole and demonstrated the importance of the use of histopathology for the diagnosis of therapeutic efficacy in field strains.

Resumo Na atualidade, o albendazol é uma das drogas mais usadas devido à sua acessibilidade econômica. O objetivo foi avaliar a histopatologia dos espécimes de Fasciola hepatica. Para isso, foi realizado o teste de eficácia em ovinos tratados com albendazol na dose recomendada para Fasciola hepatica, no qual os helmintos recuperados em necropsia foram contabilizados e separados para histologia. As células espermatogênicas de parasitas recuperados de ovinos tratados e controle foram examinadas por microscopia. O teste de redução de ovos por grama de fezes revelou 97,06% de eficácia do albendazol no tratamento de F. hepatica. As alterações nas células dos túbulos testiculares iniciaram-se 48 horas após o tratamento e tornaram-se evidentes em 72 horas, altura em que tornou-se difícil identificar os tipos de células. As espermatogônias primárias e secundárias tornaram-se cada vez mais raras e a vacuolização intercelular foi mais evidente. Sinais de apoptose, com núcleos picnóticos e evidência de cariorrexia foram observados em todos os momentos. Os detritos celulares foram identificados 96 horas após o tratamento. Os resultados indicaram que a espermatogênese parasitária foi severamente afetada pelo albendazol e demonstrou a importância do uso da histopatologia para o diagnóstico de eficácia terapêutica em cepas de campo.

Microscopic alterations in Fasciola hepatica from sheep treated with albendazole.

Currently, albendazole is one of the most commonly used drugs because of its affordability. The objective was to evaluate the histopathology of Fasciola hepatica specimens. For this, the efficacy test was performed on sheep treated with albendazole at the dose recommended for F. hepatica, in which the helminths recovered at necropsy were counted and separated for histology. Spermatogenic cells from parasites recovered from treated and control sheep were examined by microscopy. The fecal egg-count reduction test revealed 97.06% efficacy of albendazole in the treatment of F. hepatica. Changes in testicular tubule cells started 48 hours after treatment and became evident within 72 hours, at which point it became difficult to identify cell types. Primary and secondary spermatogonia became increasingly rare and intercellular vacuolization was more evident. Signs of apoptosis, with pycnotic nuclei and evidence of keriorrexia were observed at all times. Cell debris was identified 96 hours after treatment. The results indicated that parasitic spermatogenesis was severely affected by albendazole and demonstrated the importance of the use of histopathology for the diagnosis of therapeutic efficacy in field strains.

Spatial analysis on the risk of bovine cysticercosis occurrence in the state of Espírito Santo, Brazil.

A map of the risk of bovine cysticercosis occurrence was developed for the state of Espírito Santo, Brazil, and it was based in a mathematic model based on following variables: inadequate sewage, bovine population by county, use and occupation of the land and flood risks in GIS environ by means of the ArcGIS/ArcINFO 10.1 program. The work aims to spatially analyze the risk of bovine cysticercosis occurrence in the state of Espírito Santo, by means of risk factors related to cysticercosis and compare with the prevalence obtained from slaughterhouses in the same area. The map of risk showed areas high risk and very high risk located mainly in Ecoporanga, Linhares, counties, where the prevalence from slaughterhouses are low, and in two counties of south macro-region, Presidente Kennedy and Itapemirim, where prevalence from slaughterhouses are higher.

Percentual de suplementação de fonte taninífera na ração concentrada de caprinos jovens sobre o desempenho e carga parasitária / Tanniferous plant supplementation percentage in young goats concentrates diet on perfermance and parasite contro

O presente estudo buscou determinar o percentual de adição de torta de pimenta rosa (TPR) na dieta de caprinos jovens sobre o desempenho e carga parasitária. Foram utilizados 25 caprinos jovens da raça Saanen, distribuídos em um delineamento inteiramente casualizado em grupos de cinco tratamentos e cinco repetições, constituídos por grupos: não suplementados com TPR; suplementados com 15% TPR; suplementados com 30% TPR; suplementado com 45% TPR; e suplementado com 60% TPR. O aumento dos níveis de suplementação de pimenta rosa resultou em efeito quadrático sobre o consumo voluntário de matéria seca (CVMS), peso metabólico e ganho em peso diário (P≤0,05). Para o CVMS, o comportamento quadrático (P≤0,05) revelou ponto de mínimo de 67,1g kg0,75 dia-1 no percentual de 59,8% de TPR; para o peso metabólico, um ponto máximo de 8,5kg0,75 no percentual de 15,5% de TPR; e, para o ganho em peso diário, um ponto máximo de 0,107g dia-1 no percentual de 19,6% de TPR na dieta. Em relação à influencia da TPR sobre a carga parasitária, observou-se efeito quadrático dos níveis de suplementação de TPR na contagem de ovos por grama de fezes (OPG) (P≤0,05), com ponto de mínimo estimado em 1,0Log10 no nível de 35,8% TPR na dieta, representando o limite de resposta do TPR. Além da redução no OPG, houve um decréscimo linear na contagem de oocistos por grama de fezes (OOPG) com o incremento nos níveis de TPR da dieta. Conclui-se que a adição de 20-30% TPR possibilitou o controle de parasitos gastrointestinais com máximo desempenho animal.

The aim of this study was to evaluate the pink pepper supplementation (PPS) in young goats concentrate diet on performance parameters. Twenty five female Saanen goats were divided into five groups having five goats in each treatment. In a completely randomized design were given 0%, 15%, 30%, 45% and 60% of pink pepper supplementation in the diet. The increase levels of pink pepper supplementation resulted in a quadratic effect on voluntary intake of dry matter, metabolic weight and daily weight gain (P≤0.05). For voluntary intake of dry matter, the quadratic effect (P≤0.05) revealed minimum point of 67.1 (g kg0.75 dia-1) in the percentage of 59.8% PPS; for metabolic weight a peak of 8. 5kg0.75 in the percentage of 15.5% PPS, and the daily weight gain a peak of 0.107g day-1 in the percentage of 19.6% PPS in the diet. Regarding the effect of PPS on the parasite load, there was a quadratic effect of dietary supplementation of the PPS on egg count per gram of feces (P≤0.05), with minimum point estimated at 1.0log10 at 35.8% of PPS in the diet, representing the limit of PPS response. There was a linear decrease on oocyst count per gram feces with increased levels of PPS 's diet. It was concluded that the supplementation up to 20-30% of pink pepper was able to control of gastrointestinal parasites with maximum animal performance.

Distribution of bovine fasciolosis and associated factors in south Espírito Santo, Brazil: an update.

The geographical distribution and factors associated with bovine fasciolosis in the south of Espírito Santo were updated and the prevalences of this disease and of snails of the genus Lymnaea in the municipality of Jerônimo Monteiro were calculated. In the first stage, fecal samples were collected from 10% of the herds of 115 farms in 23 municipalities and interviews were conducted with owners. Generalized linear mixed models were used. In the second stage, in Jerônimo Monteiro municipality, feces and mollusks were collected from all farms registered in the milk cooperatives in the region. The mollusks were identified and examined for infection by Fasciola hepatica. Fasciolosis was diagnosed in 18 (78%) of the 23 municipalities. Of the 1157 fecal samples examined, 19.01% were positive for eggs of F. hepatica. The final model shows statistical evidence of associations between positive farms and previous cases of fasciolosis and concomitant grazing of cattle with other definitive hosts. In the evaluated farms from the studied municipality the prevalence of fasciolosis and Lymnaea was of 66.7% and 23.8%, respectively. Mollusks were found in flooded areas and the animals' drinking water troughs. The wide geographical distribution of bovine fasciolosis in the south of Espírito Santo requires control measures to prevent its expansion towards the north of this state and other places characterized as F. hepatica free-infection.

Distribution of bovine fasciolosis and associated factors in south Espírito Santo, Brazil: an update / Distribuição da fasciolose bovina e fatores associados no sul do Espírito Santo, Brasil: uma atualização

The geographical distribution and factors associated with bovine fasciolosis in the south of Espírito Santo were updated and the prevalences of this disease and of snails of the genus Lymnaea in the municipality of Jerônimo Monteiro were calculated. In the first stage, fecal samples were collected from 10% of the herds of 115 farms in 23 municipalities and interviews were conducted with owners. Generalized linear mixed models were used. In the second stage, in Jerônimo Monteiro municipality, feces and mollusks were collected from all farms registered in the milk cooperatives in the region. The mollusks were identified and examined for infection by Fasciola hepatica. Fasciolosis was diagnosed in 18 (78%) of the 23 municipalities. Of the 1157 fecal samples examined, 19.01% were positive for eggs of F. hepatica. The final model shows statistical evidence of associations between positive farms and previous cases of fasciolosis and concomitant grazing of cattle with other definitive hosts. In the evaluated farms from the studied municipality the prevalence of fasciolosis and Lymnaea was of 66.7% and 23.8%, respectively. Mollusks were found in flooded areas and the animals' drinking water troughs. The wide geographical distribution of bovine fasciolosis in the south of Espírito Santo requires control measures to prevent its expansion towards the north of this state and other places characterized as F. hepatica free-infection.

Realizou-se um estudo atualizado da distribuição geográfica e dos fatores associados à fasciolose bovina, no Sul do Estado do Espírito Santo, e calculadas a prevalência de moluscos do gênero Lymnaea no município de Jerônimo Monteiro. Na primeira etapa, coletaram-se amostras fecais de 10% dos rebanhos de 115 propriedades, em 23 municípios, e realizaram-se entrevistas com os proprietários. Modelos lineares generalizados mistos foram utilizados. Na segunda etapa, em Jerônimo Monteiro, coletaram-se fezes e moluscos em todas as propriedades cadastradas nas cooperativas de leite da região. Os moluscos foram identificados e examinados quanto à infecção por Fasciola hepatica. A fasciolose foi diagnosticada em 18 (78%) dos 23 municípios. Das 1157 amostras fecais examinadas, 19,01% foram positivas para ovos de F. hepatica. O modelo final mostra, concomitantemente, evidências estatísticas de associação entre propriedades positivas e casos anteriores de fasciolose e de pastoreio de bovinos com outros hospedeiros definitivos. Em Jerônimo Monteiro, a prevalência de fasciolose foi 66,7% e de Lymnaea 23,8%. Moluscos foram encontrados em áreas alagadas e em bebedouros dos animais. A ampla distribuição geográfica da fasciolose bovina, no Sul do Espírito Santo, requer medidas de controle que evitem sua expansão em direção ao Norte do Estado.

Kit comercial de ELISA® para a detecção de coproantígenos e exame coproparasitológico em bovinos com fígados condenados por fasciolose / Commercial ELISA® kit for detection of coproantigen and coproparasitological method in bovine livers with fascioliasis convicted

Objetivou-se com o presente estudo comparar um kit comercial de ELISA para a detecção de coproantígenos e um exame coproparasitológico de sedimentação utilizando-se como padrão ouro o diagnóstico da inspeção de fígados bovinos ao abate. Além disso, avaliou-se a correlação entre a intensidade parasitária mensurada pela contagem de ovos nas fezes e a de parasitos ao abate. Foram coletadas as fezes e avaliados macroscopicamente os fígados de 81 bovinos, dos quais 45 tiveram os fígados condenados por fasciolose hepática ao abate, nos quais se realizou a contagem dos parasitos. Duas frações de cada amostra de fezes coletadas foram separadas e uma delas foi armazenada em congelador para posterior realização do ELISA e a outra processada segundo uma técnica de sedimentação fecal para diagnóstico de ovos de Fasciola hepatica. O coeficiente de correlação de Spearman e o qui-quadrado de McNemar foram utilizados, adotando-se o nível de significância de 5%. Em oito fígados bovinos condenados por apresentarem lesões características de fasciolose, não foram encontrados exemplares do parasito. O exame coproparasitológico e o ELISA para detecção de coproantígenos, respectivamente, apresentaram sensibilidade de 51,11% e 75,55%, especificidade de 100% e 91,66%, valor preditivivo positivo de 100% e 91,89%, valor preditivo negativo de 62% e 75% e kappa de 0,48 e 0,65. Os resultados obtidos pelo kit ELISA comercial não diferiram (P=0,06) dos obtidos ao abate, mas o exame coproparasitológico diferiu (P<0,0001) do abate na detecção de animais positivos. A correlação entre o número de parasitos no fígado e o número de ovos nas fezes é moderada (rs=0,5757, P<0,0001). O kit ELISA comercial foi mais sensível do que o exame coproparasitológico, embora este não deva ser descartado devido a sua eficiência.

The aim of this study was to compare a commercial ELISA kit for detection on coproantigen examination and fecal sedimentation using as gold standard inspection diagnosis of bovine livers at slaughter. In addition, we evaluated the correlation between the measured intensity of infection by counting eggs in the feces and the parasites in bovine livers. Feces were collected and evaluated macroscopically of 81 cattle livers, 45 of which had livers condemned by liver flukes and in these livers parasites were counted. Two fractions of stool samples collected were separated and one stored in freezer for further ELISA and other one processed according to sedimentation technique for diagnosis Fasciola hepatica. The Spearman correlation and McNemar chi-square were used, adopting the significance of 5%. In eight bovine livers condemned by the characteristic lesions of fascioliasis parasite were not found. The stool examinations and ELISA testing for detection coproantigen, respectively, had sensitivity of 51.11% and 75.55%, specificity of 100% and 91,66%, predictive positive value was 100% and 91.89%, predictive negative value 62% and 75% and kappa 0.48 and 0.65. The results obtained by commercial® ELISA kit did not differ (P=0,06) obtained at slaughterhouse, but the stool examinations differed (P<0.0001) in the detection of the positive animals. The correlation between the number of parasites in the liver and the number of eggs in the feces was moderate (rs=0.5757, P<0.0001). The commercial ELISA kit® was more sensitive than the fecal test, althought this one shoud not be discarded because of their efficiency.

Application of a geographical information system approach for risk analysis of fascioliasis in southern Espírito Santo state, Brazil.

A model based on geographical information systems for mapping the risk of fascioliasis was developed for the southern part of Espírito Santo state, Brazil. The determinants investigated were precipitation, temperature, elevation, slope, soil type and land use. Weightings and grades were assigned to determinants and their categories according to their relevance with respect to fascioliasis. Theme maps depicting the spatial distribution of risk areas indicate that over 50% of southern Espírito Santo is either at high or at very high risk for fascioliasis. These areas were found to be characterized by comparatively high temperature but relatively low slope, low precipitation and low elevation corresponding to periodically flooded grasslands or soils that promote water retention.

Prevalence of liver condemnation due to bovine fasciolosis in Southern Espírito Santo: temporal distribution and economic losses.

The present study was conducted to evaluate the economic losses and temporal distribution of the prevalence of liver condemnation due to bovine fasciolosis. The abattoir in Atílio Vivácqua, in the South of the State of Espírito Santo, which is under state inspection by the veterinary service of the Livestock and Forest Protection Institute of Espírito Santo, was used as the data source. The prevalence of liver condemnation due to fasciolosis over the period 2006-2009 was calculated. The χ(2) test, simple linear regression analysis and χ(2) for trend were used, with a significance level of p ≤ 0.05. Over the period analyzed, 110,956 cattle were slaughtered and the prevalence of liver condemnation due to Fasciola hepatica was 15.24% in 2006, 23.93% in 2007, 28.57% in 2008 and 28.24% in 2009. The historical trend of liver condemnation is an increasing trend, thus indicating that this parasitism has become established in the herd as a problem in this region, with prevalence similar to that of traditionally endemic regions. Condemnations occurred throughout the year, with the highest prevalence in April and May and with significant differences between the dry and wet seasons. The economic losses from liver condemnation can be considered high.