Biological activity of cinnamaldehyde, citronellal, geraniol and anacardic acid on Haemonchus contortus isolates susceptible and resistant to synthetic anthelmintics.

Parasitism by gastrointestinal nematodes is a challenge for small ruminant farming worldwide. It causes productive and economic losses, especially due to parasite resistance to conventional anthelmintics. Natural compounds with antiparasitic activity are a potential alternative for controlling these parasites especially when considering the widespread occurrence of anthelmintic resistance. Our objective was to evaluate the activity of anacardic acid, geraniol, cinnamaldehyde and citronellal on Haemonchus contortus isolates with different levels of anthelmintic resistance profiles. These compounds were tested using egg hatch assays (EHAs), larval development tests (LDTs) as well as LDTs on mini-fecal cultures, on the Haemonchus contortus isolates Kokstad (KOK-resistant to all anthelmintics), Inbred-Strain-Edinburgh (ISE-susceptible to all anthelmintics) and Echevarria (ECH-susceptible to all anthelmintics). Effective concentrations to inhibit 50% (EC50) and 95% (EC95) of egg hatching and larval development were calculated. Results for EHA and LDT for all tested compounds, considering EC50 and EC95 values, showed low variation among the studied isolates with most RF values below 2x. All studied compounds showed efficacy against egg hatching and larval development of H. contortus isolates regardless of anthelmintic resistance profiles. The compounds with the smallest EC50 and EC95 values were cinnamaldehyde and anacardic acid making them promising candidates for future in vivo studies.

Biological activity of cinnamaldehyde, citronellal, geraniol and anacardic acid on Haemonchus contortus isolates susceptible and resistant to synthetic anthelmintics / Atividade biológica dos compostos cinamaldeído, citronelal, geraniol e ácido anacárdico em isolados de Haemonchus contortus susceptíveis e resistentes a anti-helmínticos sintéticos

Parasitism by gastrointestinal nematodes is a challenge for small ruminant farming worldwide. It causes productive and economic losses, especially due to parasite resistance to conventional anthelmintics. Natural compounds with antiparasitic activity are a potential alternative for controlling these parasites especially when considering the widespread occurrence of anthelmintic resistance. Our objective was to evaluate the activity of anacardic acid, geraniol, cinnamaldehyde and citronellal on Haemonchus contortus isolates with different levels of anthelmintic resistance profiles. These compounds were tested using egg hatch assays (EHAs), larval development tests (LDTs) as well as LDTs on mini-fecal cultures, on the Haemonchus contortus isolates Kokstad (KOK-resistant to all anthelmintics), Inbred-Strain-Edinburgh (ISE-susceptible to all anthelmintics) and Echevarria (ECH-susceptible to all anthelmintics). Effective concentrations to inhibit 50% (EC50) and 95% (EC95) of egg hatching and larval development were calculated. Results for EHA and LDT for all tested compounds, considering EC50 and EC95 values, showed low variation among the studied isolates with most RF values below 2x. All studied compounds showed efficacy against egg hatching and larval development of H. contortus isolates regardless of anthelmintic resistance profiles. The compounds with the smallest EC50 and EC95 values were cinnamaldehyde and anacardic acid making them promising candidates for future in vivo studies.(AU)

A infecção por nematoides gastrintestinais é um dos principais desafios na produção de pequenos ruminantes e ocasiona perdas produtivas, principalmente, devido à resistência anti-helmíntica. Bioativos com atividade anti-helmíntica são potencial alternativa para o controle desses parasitos em especial, considerando-se a ampla incidência de resistência anti-helmíntica nos rebanhos. O objetivo deste estudo foi avaliar a atividade biológica do ácido anacárdico, geraniol, cinamaldeído e citronelal em isolados de Haemonchus contortus com diferentes perfis de resistência anti-helmíntica. Foram realizados testes de eclosão de ovos (TEO), testes de desenvolvimento larvar (TDL) e TDLs em minicoproculturas, utilizando-se o isolado Kokstad (resistente a todos os anti-helmínticos), o isolado Inbred-Strain-Edinburgh (suscetível) e o isolado Echevarria (suscetível). Foram calculadas as concentrações efetivas para inibir 50% (CE50) e 95% (CE95) da eclodibilidade dos ovos e do desenvolvimento larvar. Resultados de TEO e TDL apresentaram baixa variação entre os diferentes isolados para um mesmo composto testado com fatores de resistência geralmente abaixo de 2x. Todos os compostos estudados mostraram eficácia contra a eclosão de ovos e desenvolvimento larvar de isolados de H. contortus independente do perfil de resistência anti-helmíntica dos mesmos. Os compostos que apresentaram atividade nas menores concentrações foram cinamaldeído e ácido anacárdico, sendo estes os componentes mais promissores para futuros estudos in vivo.(AU)

Castor cake as organic fertilizer to control gastrointestinal nematodes in pasture-raised sheep.

Gastrointestinal parasitism is one of the factors that discourages farmers from raising small ruminants in cultivated pastures. To validate a soil treatment strategy to control the free-living stages of gastrointestinal nematodes (GIN), castor cake (CC) was used as a fertilizer on a pasture where sheep grazed on guinea grass under continuous stocking. On day zero, the pasture was divided into three paddocks, contaminated by GIN and treated, respectively, with CC divided into two applications (2CC1/2), CC in a single application (CC1) and organic compost in a single application (control). On day 21, eight GIN-free sheep were placed in each paddock. On day 58, significant differences (P<0.05) were observed: reduction of up to 66.10% in larvae.g-1 of dry mass in pastures fertilized with CC, decrease of up to 60.72% in infection rates among the animals in the groups treated with CC, higher average daily weight gain (over 185 g.day-1) and packed cell volume (over 26%) in the groups treated with CC, when compared to the control (128 g.day-1; 20.9%). In view of the results, the use of CC, mainly CC1, as a fertilizer for guinea grass pastures, under continuous stocking, proved to be promising, with 63.41% effectiveness in controlling worm infestations.

Castor cake as organic fertilizer to control gastrointestinal nematodes in pasture-raised sheep / Torta de mamona como adubo orgânico para o controle de nematoides gastrintestinais em ovinos criados a pasto

Gastrointestinal parasitism is one of the factors that discourages farmers from raising small ruminants in cultivated pastures. To validate a soil treatment strategy to control the free-living stages of gastrointestinal nematodes (GIN), castor cake (CC) was used as a fertilizer on a pasture where sheep grazed on guinea grass under continuous stocking. On day zero, the pasture was divided into three paddocks, contaminated by GIN and treated, respectively, with CC divided into two applications (2CC1/2), CC in a single application (CC1) and organic compost in a single application (control). On day 21, eight GIN-free sheep were placed in each paddock. On day 58, significant differences (P 0.05) were observed: reduction of up to 66.10% in larvae.g-1 of dry mass in pastures fertilized with CC, decrease of up to 60.72% in infection rates among the animals in the groups treated with CC, higher average daily weight gain (over 185 g.day-1) and packed cell volume (over 26%) in the groups treated with CC, when compared to the control (128 g.day-1; 20.9%). In view of the results, the use of CC, mainly CC1, as a fertilizer for guinea grass pastures, under continuous stocking, proved to be promising, with 63.41% effectiveness in controlling worm infestations.(AU)

O parasitismo gastrintestinal é um dos fatores que fragiliza a exploração de pequenos ruminantes em pastagens cultivadas. Objetivando validar a estratégia de tratamento do solo para o controle dos estágios de vida livre de nematoides gastrintestinais (NGI), a torta de mamona (TM) foi utilizada como adubo, com ovinos pastejando em capim-tanzânia sob lotação contínua. No dia zero, o pasto foi dividido em três piquetes, contaminados por NGI e tratados, respectivamente, com TM parcelada em duas aplicações (2TM1/2), TM em uma única aplicação (TM1) e composto orgânico em única aplicação (testemunha). No dia 21, cada piquete recebeu oito ovinos livres de NGI. No dia 58, observaram-se diferenças significativas (P 0,05): redução de até 66,10% de larvas.g-1 de massa seca nas pastagens adubadas com TM; redução de até 60,72% da infecção dos animais nos grupos tratados com TM; ganho de peso médio diário (acima de 185 g.dia-1) e volume globular (acima de 26%) superior nos grupos tratados com TM, quando comparados com a testemunha (128 g.dia-1; 20,9%). Diante dos resultados, o uso da TM, principalmente TM1, como adubo em pasto de capim-tanzânia, sob lotação contínua, mostrou-se promissor, com eficácia de 63,41% para controlar a verminose.(AU)

Castor cake as organic fertilizer to control gastrointestinal nematodes in pasture-raised sheep / Torta de mamona como adubo orgânico para o controle de nematoides gastrintestinais em ovinos criados a pasto

Abstract Gastrointestinal parasitism is one of the factors that discourages farmers from raising small ruminants in cultivated pastures. To validate a soil treatment strategy to control the free-living stages of gastrointestinal nematodes (GIN), castor cake (CC) was used as a fertilizer on a pasture where sheep grazed on guinea grass under continuous stocking. On day zero, the pasture was divided into three paddocks, contaminated by GIN and treated, respectively, with CC divided into two applications (2CC1/2), CC in a single application (CC1) and organic compost in a single application (control). On day 21, eight GIN-free sheep were placed in each paddock. On day 58, significant differences (P<0.05) were observed: reduction of up to 66.10% in larvae.g-1 of dry mass in pastures fertilized with CC, decrease of up to 60.72% in infection rates among the animals in the groups treated with CC, higher average daily weight gain (over 185 g.day-1) and packed cell volume (over 26%) in the groups treated with CC, when compared to the control (128 g.day-1; 20.9%). In view of the results, the use of CC, mainly CC1, as a fertilizer for guinea grass pastures, under continuous stocking, proved to be promising, with 63.41% effectiveness in controlling worm infestations.

Resumo O parasitismo gastrintestinal é um dos fatores que fragiliza a exploração de pequenos ruminantes em pastagens cultivadas. Objetivando validar a estratégia de tratamento do solo para o controle dos estágios de vida livre de nematoides gastrintestinais (NGI), a torta de mamona (TM) foi utilizada como adubo, com ovinos pastejando em capim-tanzânia sob lotação contínua. No dia zero, o pasto foi dividido em três piquetes, contaminados por NGI e tratados, respectivamente, com TM parcelada em duas aplicações (2TM1/2), TM em uma única aplicação (TM1) e composto orgânico em única aplicação (testemunha). No dia 21, cada piquete recebeu oito ovinos livres de NGI. No dia 58, observaram-se diferenças significativas (P<0,05): redução de até 66,10% de larvas.g-1 de massa seca nas pastagens adubadas com TM; redução de até 60,72% da infecção dos animais nos grupos tratados com TM; ganho de peso médio diário (acima de 185 g.dia-1) e volume globular (acima de 26%) superior nos grupos tratados com TM, quando comparados com a testemunha (128 g.dia-1; 20,9%). Diante dos resultados, o uso da TM, principalmente TM1, como adubo em pasto de capim-tanzânia, sob lotação contínua, mostrou-se promissor, com eficácia de 63,41% para controlar a verminose.

An epidemiological study of caprine arthritis encephalitis virus (CAEV) in breeder goats from Northeastern Brazil / Estudo epidemiológico da Artrite Encefalite Caprina á vírus (CAEV) com ênfase em reprodutores de rebanhos do Nordeste do Brasil

Caprine arthritis encephalitis virus (CAEV) is a retrovirus that infects goats. This study evaluated the prevalence of CAEV in breeder goats from the states of Maranhão, Ceará, Piauí, Alagoas, Sergipe, Rio Grande do Norte, and Paraíba. We collected a total of 531 serum samples from 251 properties. On average, two male breeder goats were examined from each farm. Results from western blotting demonstrated that the prevalence of CAEV was 6.2% (32/513). In each state, the following prevalence values were found: Piauí, 5.9% (7/119); Maranhão, 2.0% (01/48); Sergipe, 7.1% (03/42); Alagoas, 17.6% (03/17); Rio Grande do Norte, 4.7% (05/105); Paraíba, 2.1% (02/94); and Ceará, 12.5% (11/34). We also conducted a univariate analysis to determine the risk factors that are associated with CAEV. This analysis revealed that breeding season, records of herd data, criteria adopted for the first mating of females, castration of male goats, origin of breeders, and identification of the animal were associated with CAEV. Adopting control measures to identify CAEV-positive animals and avoid virus transmission to females, especially during breeding seasons, is crucial since, males carrying CAEV can be sources of infection for the entire herd.(AU)

Objetivou-se com esse estudo avaliar a prevalência da Artrite Encefalite Caprina (CAE) em reprodutores dos estados do Maranhão, Ceará, Piauí, Alagoas, Sergipe, Rio Grande do Norte e Paraíba. Para tanto, foram examinadas em média dois reprodutores por criatório, totalizando 513 amostras de soros e 251 propriedades. A prevalência encontrada através do Western Blotting foi de 6,2% (32/513). Em cada estado participante do estudo foram encontradas as prevalências descritas a seguir: Piauí 5,9% (7/119), Maranhão 2,0% (01/48), Sergipe 7,1% (03/42), Alagoas 17,6% (03/17), Rio Grande do Norte 4,7% (05/105), Paraíba 2,1% (02/94) e Ceará 12,5% (11/34). Na análise univariável para os fatores de risco, as variáveis associadas (p ≤ 0,20) a frequência de positividade nos reprodutores foi: estação de monta, anotações em relação ao rebanho, critério adotado para a primeira cobertura das fêmeas, castração dos caprinos machos, origem dos reprodutores e identificação dos animais. Na análise de regressão logística múltipla, não foram encontrados fatores de risco para a infecção em estudo.(AU)

An epidemiological study of caprine arthritis encephalitis virus (CAEV) in breeder goats from Northeastern Brazil / Estudo epidemiológico da Artrite Encefalite Caprina á vírus (CAEV) com ênfase em reprodutores de rebanhos do Nordeste do Brasil

Caprine arthritis encephalitis virus (CAEV) is a retrovirus that infects goats. This study evaluated the prevalence of CAEV in breeder goats from the states of Maranhão, Ceará, Piauí, Alagoas, Sergipe, Rio Grande do Norte, and Paraíba. We collected a total of 531 serum samples from 251 properties. On average, two male breeder goats were examined from each farm. Results from western blotting demonstrated that the prevalence of CAEV was 6.2% (32/513). In each state, the following prevalence values were found: Piauí, 5.9% (7/119); Maranhão, 2.0% (01/48); Sergipe, 7.1% (03/42); Alagoas, 17.6% (03/17); Rio Grande do Norte, 4.7% (05/105); Paraíba, 2.1% (02/94); and Ceará, 12.5% (11/34). We also conducted a univariate analysis to determine the risk factors that are associated with CAEV. This analysis revealed that breeding season, records of herd data, criteria adopted for the first mating of females, castration of male goats, origin of breeders, and identification of the animal were associated with CAEV. Adopting control measures to identify CAEV-positive animals and avoid virus transmission to females, especially during breeding seasons, is crucial since, males carrying CAEV can be sources of infection for the entire herd.

Objetivou-se com esse estudo avaliar a prevalência da Artrite Encefalite Caprina (CAE) em reprodutores dos estados do Maranhão, Ceará, Piauí, Alagoas, Sergipe, Rio Grande do Norte e Paraíba. Para tanto, foram examinadas em média dois reprodutores por criatório, totalizando 513 amostras de soros e 251 propriedades. A prevalência encontrada através do Western Blotting foi de 6,2% (32/513). Em cada estado participante do estudo foram encontradas as prevalências descritas a seguir: Piauí 5,9% (7/119), Maranhão 2,0% (01/48), Sergipe 7,1% (03/42), Alagoas 17,6% (03/17), Rio Grande do Norte 4,7% (05/105), Paraíba 2,1% (02/94) e Ceará 12,5% (11/34). Na análise univariável para os fatores de risco, as variáveis associadas (p ≤ 0,20) a frequência de positividade nos reprodutores foi: estação de monta, anotações em relação ao rebanho, critério adotado para a primeira cobertura das fêmeas, castração dos caprinos machos, origem dos reprodutores e identificação dos animais. Na análise de regressão logística múltipla, não foram encontrados fatores de risco para a infecção em estudo.

Crop residues activity against the free-living stages of small ruminant nematodes.

The nematicidal effect of different organic materials was evaluated in order to develop a non-chemical alternative soil treatment for control of the free-living stages of small ruminant gastrointestinal nematodes. The selected organic materials were residues from the juice industry of acerola, cashew, grape, guava, papaya and pineapple, as well as castor residue from the biodiesel industry. LC90 results showed that pineapple residue was the most efficient inhibitor of larval development, followed by castor, grape, cashew, acerola, guava and papaya. Castor residue was also a good source of nitrogen and was used in a greenhouse experiment to prevent larval development in contaminated goat faeces that was deposited in pots containing the grasses Brachiaria brizantha (var. Paiaguás) or Megathyrsus maximus x M. infestum (var. Massai). Castor residue caused a significant (P < 0.05) reduction (85.04%) in Paiaguás grass contamination (L3.dry mass-1) and a reduction of 17.35% in Massai grass contamination (P > 0.05), with an increase in the biomass production of Massai (251.43%, P < 0.05) and Paiaguás (109.19%, P > 0.05) grasses. This strategy, called Econemat®, with good results in vitro shows to be promising on pasture increasing phytomass production.

Crop residues activity against the free-living stages of small ruminant nematodes / Atividade de resíduos de culturas contra os estágios de vida livre de nematoides de pequenos ruminantes

Abstract The nematicidal effect of different organic materials was evaluated in order to develop a non-chemical alternative soil treatment for control of the free-living stages of small ruminant gastrointestinal nematodes. The selected organic materials were residues from the juice industry of acerola, cashew, grape, guava, papaya and pineapple, as well as castor residue from the biodiesel industry. LC90 results showed that pineapple residue was the most efficient inhibitor of larval development, followed by castor, grape, cashew, acerola, guava and papaya. Castor residue was also a good source of nitrogen and was used in a greenhouse experiment to prevent larval development in contaminated goat faeces that was deposited in pots containing the grasses Brachiaria brizantha (var. Paiaguás) or Megathyrsus maximus x M. infestum (var. Massai). Castor residue caused a significant (P < 0.05) reduction (85.04%) in Paiaguás grass contamination (L3.dry mass-1) and a reduction of 17.35% in Massai grass contamination (P > 0.05), with an increase in the biomass production of Massai (251.43%, P < 0.05) and Paiaguás (109.19%, P > 0.05) grasses. This strategy, called Econemat®, with good results in vitro shows to be promising on pasture increasing phytomass production.

Resumo O efeito nematicida de diferentes materiais orgânicos foi avaliado, com o objetivo de desenvolver um tratamento alternativo não químico do solo para o controle dos estágios de vida livre de nematoides gastrintestinais de pequenos ruminantes. Os materiais orgânicos selecionados foram resíduos da indústria de suco de acerola, caju, uva, goiaba, mamão e abacaxi, além do resíduo de mamona da indústria de biodiesel. Os resultados da LC90 mostraram que o resíduo de abacaxi foi o mais eficiente inibidor do desenvolvimento larval, seguido pela mamona, uva, caju, acerola, goiaba e mamão. O resíduo de mamona também se mostrou uma boa fonte de nitrogênio, sendo usado em experimento em casa de vegetação para prevenir o desenvolvimento larval em fezes contaminadas, depositadas em vasos, contendo as gramíneas Brachiaria brizantha (var. Paiaguás) ou Megathyrsus maximus x M. infestum (var. Massai). O resíduo de mamona reduziu (85,04%) significativamente (P < 0,05) a contaminação no capim-Paiaguás (L3.dry massa-1), 17,35% no capim Massai (P> 0,05), e aumentou a produção de biomassa das gramíneas Massai (251,43%, P <0,05) e Paiaguás (109,19%, P> 0,05). Essa estratégia, chamada Econemat®, com bons resultados in vitro, mostra-se promissora nas pastagens aumentando a produção de fitomassa.

Crop residues activity against the free-living stages of small ruminant nematodes / Atividade de resíduos de culturas contra os estágios de vida livre de nematoides de pequenos ruminantes

The nematicidal effect of different organic materials was evaluated in order to develop a non-chemical alternative soil treatment for control of the free-living stages of small ruminant gastrointestinal nematodes. The selected organic materials were residues from the juice industry of acerola, cashew, grape, guava, papaya and pineapple, as well as castor residue from the biodiesel industry. LC90 results showed that pineapple residue was the most efficient inhibitor of larval development, followed by castor, grape, cashew, acerola, guava and papaya. Castor residue was also a good source of nitrogen and was used in a greenhouse experiment to prevent larval development in contaminated goat faeces that was deposited in pots containing the grasses Brachiaria brizantha (var. Paiaguás) or Megathyrsus maximus x M. infestum (var. Massai). Castor residue caused a significant (P 0.05) reduction (85.04%) in Paiaguás grass contamination (L3.dry mass-1) and a reduction of 17.35% in Massai grass contamination (P > 0.05), with an increase in the biomass production of Massai (251.43%, P 0.05) and Paiaguás (109.19%, P > 0.05) grasses. This strategy, called Econemat®, with good results in vitro shows to be promising on pasture increasing phytomass production.(AU)

O efeito nematicida de diferentes materiais orgânicos foi avaliado, com o objetivo de desenvolver um tratamento alternativo não químico do solo para o controle dos estágios de vida livre de nematoides gastrintestinais de pequenos ruminantes. Os materiais orgânicos selecionados foram resíduos da indústria de suco de acerola, caju, uva, goiaba, mamão e abacaxi, além do resíduo de mamona da indústria de biodiesel. Os resultados da LC90 mostraram que o resíduo de abacaxi foi o mais eficiente inibidor do desenvolvimento larval, seguido pela mamona, uva, caju, acerola, goiaba e mamão. O resíduo de mamona também se mostrou uma boa fonte de nitrogênio, sendo usado em experimento em casa de vegetação para prevenir o desenvolvimento larval em fezes contaminadas, depositadas em vasos, contendo as gramíneas Brachiaria brizantha (var. Paiaguás) ou Megathyrsus maximus x M. infestum (var. Massai). O resíduo de mamona reduziu (85,04%) significativamente (P 0,05) a contaminação no capim-Paiaguás (L3.dry massa-1), 17,35% no capim Massai (P> 0,05), e aumentou a produção de biomassa das gramíneas Massai (251,43%, P 0,05) e Paiaguás (109,19%, P> 0,05). Essa estratégia, chamada Econemat®, com bons resultados in vitro, mostra-se promissora nas pastagens aumentando a produção de fitomassa.(AU)

Haemonchus contortus ß-tubulin isotype 1 gene F200Y and F167Y SNPs are both selected by ivermectin and oxfendazole treatments with differing impacts on anthelmintic resistance.

Parasitism by Haemonchus contortus is one of the main limiting factors in small ruminant production in tropical areas. Benzimidazoles (BZ) and macrocyclic lactones (ML) are the most used anthelmintic classes in gastrointestinal nematodes control. There is considerable scientific evidence of a possible relation between the anthelmintic resistance to BZ and ML. This study aimed to characterize the dynamics of anthelmintic resistance in an H. contortus susceptible isolate under selection pressure for BZ and ML alone or in combination and the role of isotype 1 ß-tubulin gene SNPs in these situations. A total of 12 Somali sheep were infected with 5000 third stage larvae of H. contortus Inbred-Susceptible Edinburgh (ISE) isolate. Once infection was established, animals were distributed in three groups (n=4), each treated with crescent doses of oxfendazole (OXF), ivermectin (IVM) and oxfendazole plus ivermectin (IVMOXF). An additional control group with untreated animals was maintained during the entire experiment. After each treatment, eggs were collected and real-time PCR was performed to identify single nucleotide polymorphisms (SNPs) F167Y, F200Y and E198A, in addition to egg hatch test (EHT) for BZ and larval development test (LDT) for ivermectin resistance. All treatments led to increased resistance allelic frequencies at SNPs F200Y and F167Y (p <0.05). In vitro results showed increased phenotypic resistance against both anthelmintic classes in groups IVM and IVMOXF while group OXF only developed resistance against BZ. Finally, we provide evidence that while isotype 1 ß-tubulin gene SNPs may have some involvement with ML resistance, the presence of these ß-tubulin SNPs alone are not sufficient to develop ML resistance.

Ocorrência da infecção por Cryptosporidium spp. em cabritos (Capra hircus) / Occurrence of infection by Cryptosporidium spp. in goat kids (Capra hircus)

O presente estudo teve como objetivo determinar a ocorrência da infecção por Cryptosporidium spp. em cabritos de Quixadá, Ceará, Brasil. Participaram do estudo 400 cabritos, com idade entre três e 360 dias, de ambos os sexos, com e sem padrão racial definido, procedentes de 25 estabelecimentos rurais distribuídos em três circuitos. As fezes foram cadastradas de acordo com o aspecto e cor, distribuídas em tubos tipo "eppendorf®" e congeladas in natura a -20°C, até o momento das extrações de DNA genômico do parasito com auxílio de kit comercial. Para amplificação de fragmentos da subunidade 18S do RNA ribossômico (rRNA) foi utilizada a "Nested"-PCR. A ocorrência de Cryptosporidium spp em cabritos de Quixadá foi de 7,50% (30/400). A frequência no período seco e no chuvoso foi de 9,55% (19/199) e 5,47% (11/201), respectivamente (χ²=2,39 e P>0,05). Amostras positivas foram identificadas em 64,00% (16/25) das propriedades estudadas e dessas amostras 50,00% (15/30) e 70,00% (21/30) tinham as fezes com aspecto e cor normais, respectivamente, sugerindo que cabritos assintomáticos estão eliminando oocistos. Não foi observada positividade para Cryptosporidium spp. em animais com 301 a 360 dias, demonstrando que animais mais velhos apresentam menos possibilidade de se infectarem com o parasito.(AU)

The present study aimed to determine the occurrence of infection by Cryptosporidium spp. in goat kids from Quixadá, Ceará, Brazil. The study included 400 goat kids of both sexes, 3 to 360 days old, with or without defined breed, originating from 25 farms distributed in three circuits. Feces were registered in accordance with the appearance and color, distributed into tubes Eppendorf tubes and frozen in natura at-20°C until the moment of extraction of genomic DNA from the parasite with the aid of a commercial kit. For amplification of fragments of the 18S subunit of ribosomal RNA (rRNA) was used to Nested PCR. The occurrence of Cryptosporidium spp. in goats kids of the Quixadá was 7.50% (30/400). The frequency in the dry period and rainy was 9.55% (19/199) and 5.47% (11/201) respectively (χ²=2.39 and P>0.05). Positive samples were identified in 64.00% (16/25) of the studied farms, and from these samples 50.00% (15/30) and 70.00% (21/30) had feces with normal appearance and color respectively, suggesting that the asymptomatic goats were eliminating oocysts. No positivity for Cryptosporidium spp. was observed in 301 to 360-day-old goats, demonstrating that older animals have less chance to become infected with the parasite.(AU)

Ocorrência da infecção por Cryptosporidium spp. em cabritos (Capra hircus) / Occurrence of infection by Cryptosporidium spp. in goat kids (Capra hircus)

O presente estudo teve como objetivo determinar a ocorrência da infecção por Cryptosporidium spp. em cabritos de Quixadá, Ceará, Brasil. Participaram do estudo 400 cabritos, com idade entre três e 360 dias, de ambos os sexos, com e sem padrão racial definido, procedentes de 25 estabelecimentos rurais distribuídos em três circuitos. As fezes foram cadastradas de acordo com o aspecto e cor, distribuídas em tubos tipo "eppendorf®" e congeladas in natura a -20°C, até o momento das extrações de DNA genômico do parasito com auxílio de kit comercial. Para amplificação de fragmentos da subunidade 18S do RNA ribossômico (rRNA) foi utilizada a "Nested"-PCR. A ocorrência de Cryptosporidium spp em cabritos de Quixadá foi de 7,50% (30/400). A frequência no período seco e no chuvoso foi de 9,55% (19/199) e 5,47% (11/201), respectivamente (χ²=2,39 e P>0,05). Amostras positivas foram identificadas em 64,00% (16/25) das propriedades estudadas e dessas amostras 50,00% (15/30) e 70,00% (21/30) tinham as fezes com aspecto e cor normais, respectivamente, sugerindo que cabritos assintomáticos estão eliminando oocistos. Não foi observada positividade para Cryptosporidium spp. em animais com 301 a 360 dias, demonstrando que animais mais velhos apresentam menos possibilidade de se infectarem com o parasito...

The present study aimed to determine the occurrence of infection by Cryptosporidium spp. in goat kids from Quixadá, Ceará, Brazil. The study included 400 goat kids of both sexes, 3 to 360 days old, with or without defined breed, originating from 25 farms distributed in three circuits. Feces were registered in accordance with the appearance and color, distributed into tubes Eppendorf tubes and frozen in natura at-20°C until the moment of extraction of genomic DNA from the parasite with the aid of a commercial kit. For amplification of fragments of the 18S subunit of ribosomal RNA (rRNA) was used to Nested PCR. The occurrence of Cryptosporidium spp. in goats kids of the Quixadá was 7.50% (30/400). The frequency in the dry period and rainy was 9.55% (19/199) and 5.47% (11/201) respectively (χ²=2.39 and P>0.05). Positive samples were identified in 64.00% (16/25) of the studied farms, and from these samples 50.00% (15/30) and 70.00% (21/30) had feces with normal appearance and color respectively, suggesting that the asymptomatic goats were eliminating oocysts. No positivity for Cryptosporidium spp. was observed in 301 to 360-day-old goats, demonstrating that older animals have less chance to become infected with the parasite...

Lectin, hemolysin and protease inhibitors in seed fractions with ovicidal activity against Haemonchus contortus.

Bioactive molecules of plant species are promising alternatives for the chemical control of gastrointestinal nematodes in ruminants. Extracts of native and exotic seed species from Brazil's semi-arid region were tested in vitro in an egg hatch assay and the bioactivity of their proteins was investigated. Each seed species was subjected to three extractions with three types of solvents. All the seeds showed ovicidal activity, which varied according to the solvents. Higher ovicidal activity was found in the molecule fractions of low molecular weight (<12 kDa) for Albizia lebbeck, Ipomoea asarifolia, Jatropha curcas, Libidibia ferrea, Moringa oleifera and Ricinus communis (P<0.05, Bonferroni test). The two fractions of Crotalaria spectabilis showed the same ovicidal activity (P>0.05, Bonferroni test). Hemagglutinating activity was detected in the fractions of C. spectabilis and M. oleifera fractions, hemolysin activity in the A. lebbeck and M. oleifera fractions, serine protease inhibitory activity in the A. lebbeck, I. asarifolia, J. curcas, M. oleifera and R. communis fractions, cysteine protease inhibitor activity in the M. oleifera fraction, and no protein activity in the L. ferrea fraction. The results of this work reveal new plant species with a potential for use in controlling nematode parasites in goats, thus opening a new field of research involving plant protein molecules with ovicidal properties.

Lectin, hemolysin and protease inhibitors in seed fractions with ovicidal activity against Haemonchus contortus / Lectina, hemolisina e inibidores de protease em frações de sementes com atividade ovicida contra Haemonchus contortus

Bioactive molecules of plant species are promising alternatives for the chemical control of gastrointestinal nematodes in ruminants. Extracts of native and exotic seed species from Brazil's semi-arid region were tested in vitro in an egg hatch assay and the bioactivity of their proteins was investigated. Each seed species was subjected to three extractions with three types of solvents. All the seeds showed ovicidal activity, which varied according to the solvents. Higher ovicidal activity was found in the molecule fractions of low molecular weight (<12 kDa) for Albizia lebbeck, Ipomoea asarifolia, Jatropha curcas, Libidibia ferrea, Moringa oleifera and Ricinus communis (P<0.05, Bonferroni test). The two fractions of Crotalaria spectabilis showed the same ovicidal activity (P>0.05, Bonferroni test). Hemagglutinating activity was detected in the fractions of C. spectabilis and M. oleifera fractions, hemolysin activity in the A. lebbeck and M. oleifera fractions, serine protease inhibitory activity in the A. lebbeck, I. asarifolia, J. curcas, M. oleifera and R. communis fractions, cysteine protease inhibitor activity in the M. oleifera fraction, and no protein activity in the L. ferrea fraction. The results of this work reveal new plant species with a potential for use in controlling nematode parasites in goats, thus opening a new field of research involving plant protein molecules with ovicidal properties.

Moléculas bioativas de espécies vegetais são alternativas promissoras ao controle químico dos nematoides gastrintestinais em ruminantes. Extratos de sementes de espécies nativas e exóticas do Semiárido Brasileiro foram testados in vitro em ensaio de eclosão de ovos e investigada a natureza proteica da bioatividade. Três extrações com três solventes foram feitas para cada semente estudada. Todas as sementes apresentaram atividade ovicida, variando com o solvente utilizado. Maior taxa de inibição da eclosão concentrou-se nas frações de moléculas de baixa massa molecular (<12 kDa) para Albizia lebbeck, Ipomoea asarifolia, Jatropha curcas, Libidibia ferrea, Moringa oleifera e Ricinus communis (P<0,05, teste de Bonferroni). Crotalaria spectabilis mostrou atividade nas duas frações, sem diferença entre elas (P>0,05, teste de Bonferroni). Observou-se atividade hemaglutinante nas frações de C. spectabilis e M. oleifera, de hemolisina em A. lebbeck e M. oleifera, de atividade inibidora de protease da serina em A. lebbeck, I. asarifolia, J. curcas, M. oleifera e R. communis, de atividade inibidora de protease da cisteína em M. oleifera e nenhuma atividade proteica na fração de L. ferrea. Os resultados revelaram novas espécies botânicas com potencial de controle de nematoides em caprinos e um novo campo de pesquisa, o estudo de moléculas de origem proteica com atividade ovicida.

Lectin, hemolysin and protease inhibitors in seed fractions with ovicidal activity against Haemonchus contortus / Lectina, hemolisina e inibidores de protease em frações de sementes com atividade ovicida contra Haemonchus contortus

Bioactive molecules of plant species are promising alternatives for the chemical control of gastrointestinal nematodes in ruminants. Extracts of native and exotic seed species from Brazil's semi-arid region were tested in vitro in an egg hatch assay and the bioactivity of their proteins was investigated. Each seed species was subjected to three extractions with three types of solvents. All the seeds showed ovicidal activity, which varied according to the solvents. Higher ovicidal activity was found in the molecule fractions of low molecular weight (<12 kDa) for Albizia lebbeck, Ipomoea asarifolia, Jatropha curcas, Libidibia ferrea, Moringa oleifera and Ricinus communis (P<0.05, Bonferroni test). The two fractions of Crotalaria spectabilis showed the same ovicidal activity (P>0.05, Bonferroni test). Hemagglutinating activity was detected in the fractions of C. spectabilis and M. oleifera fractions, hemolysin activity in the A. lebbeck and M. oleifera fractions, serine protease inhibitory activity in the A. lebbeck, I. asarifolia, J. curcas, M. oleifera and R. communis fractions, cysteine protease inhibitor activity in the M. oleifera fraction, and no protein activity in the L. ferrea fraction. The results of this work reveal new plant species with a potential for use in controlling nematode parasites in goats, thus opening a new field of research involving plant protein molecules with ovicidal properties.

Moléculas bioativas de espécies vegetais são alternativas promissoras ao controle químico dos nematoides gastrintestinais em ruminantes. Extratos de sementes de espécies nativas e exóticas do Semiárido Brasileiro foram testados in vitro em ensaio de eclosão de ovos e investigada a natureza proteica da bioatividade. Três extrações com três solventes foram feitas para cada semente estudada. Todas as sementes apresentaram atividade ovicida, variando com o solvente utilizado. Maior taxa de inibição da eclosão concentrou-se nas frações de moléculas de baixa massa molecular (<12 kDa) para Albizia lebbeck, Ipomoea asarifolia, Jatropha curcas, Libidibia ferrea, Moringa oleifera e Ricinus communis (P<0,05, teste de Bonferroni). Crotalaria spectabilis mostrou atividade nas duas frações, sem diferença entre elas (P>0,05, teste de Bonferroni). Observou-se atividade hemaglutinante nas frações de C. spectabilis e M. oleifera, de hemolisina em A. lebbeck e M. oleifera, de atividade inibidora de protease da serina em A. lebbeck, I. asarifolia, J. curcas, M. oleifera e R. communis, de atividade inibidora de protease da cisteína em M. oleifera e nenhuma atividade proteica na fração de L. ferrea. Os resultados revelaram novas espécies botânicas com potencial de controle de nematoides em caprinos e um novo campo de pesquisa, o estudo de moléculas de origem proteica com atividade ovicida.

Identification and quantification of benzimidazole resistance polymorphisms in Haemonchus contortus isolated in Northeastern Brazil.

Haemonchus contortus is the most prevalent nematode in Brazil. The objective of this study was to select 6 populations of H. contortus of known or suspected benzimidazole resistance status and characterize these using quantitative real-time polymerase chain reaction (qPCR) for single nucleotide polymorphisms (SNPs) F200Y, F167Y and E198A in the ß-tubulin isotype 1 gene. qPCR was performed using DNA from a pool of 10 adult male H. contortus from a single animal per farm. Faecal egg count reduction test (FECRT) and egg hatch test (EHT) were used to determine the resistance status. Samples were obtained from 6 farms located in 5 counties in the Ceará State: Tauá, Boa Viagem, Quixadá, Santa Quitéria and Solonópole. The inbred-susceptible-Edinburgh (ISE) isolate was used as reference for comparative purposes in the qPCR. Benzimidazole resistance was detected by FECRT on all farms with efficacy values ranging from 0 to 51%. EC50 values as determined by EHT were all above 1.49µg/ml. High frequencies of the resistant SNPs F200Y and F167Y alleles were detected but no resistance was detected at SNP E198A. Our results suggest that the SNPs F167Y and F200Y are both important for benzimidazole resistance in the studied populations.

Espécies de Eimeria sp. em matrizes caprinas leiteiras no município de Afonso Bezerra-RN / Species of Eimeria sp. in matrix goat milk in the city of Afonso Bezerra-RN

As parasitoses gastrintestinais têm se constituído em um sério obstáculo à caprinocultura, com ênfase para a eimeriose que vem se tornando um fator limitante para a exploração desses animais. Desse modo realizou-se um estudo no período de 12 meses, com objetivo de identificar às principais espécies de Eimeria em matrizes caprinas exploradas para produção leiteira no município de Afonso Bezerra, estado do Rio Grande do Norte, determinando a porcentagem de parasitismo das espécies em duas categorias produtivas do rebanho (jovens e adultos). As fezes foram coletadas diretamente da ampola retal e enviadas ao Laboratório de Parasitologia Animal da UFERSA, onde foram processadas e identificadas, com a análise estatística feita através do SAS. Durante o período estudado, tanto os caprinos jovens quanto os adultas apresentaram positividade em relação à presença de Eimeria, com frequências de 62,2% e 37,8%, respectivamente. No que concernem às espécies encontradas, observamos que nos jovens a Eimeria ninakohlyakimova e foi a que apresentou uma maior frequência (28,42%), distintamente dos adultos, onde se destacou a E. apsheronica(26,66%). Ademais, também foram identificadas aE. alijevi, E. arloingi, E. hirci, E. jolchijevi, E.christenseni, E. caprovina eE. caprina.

Regarding the aspect of health, gastrointestinal parasites have proved to be a serious obstacle to the goat, with emphasis on eimeriosis which has become a limiting factor for the exploitation of these animals. Thus, a study was realized in 12 month, with objective to identify the main species of Eimeriain goats arrays explored for production milk in municipality of Afonso Bezerra, Rio Grande do Norte, determining the percentage of parasitism of the species into two categories of productive herd (young and adults). Feces were collected directly from the rectum and sent to the Laboratory of Animal Parasitology UFERSA, where they were processed and identified, with the statistical analysis performed using the SAS. During the study period, both the young female goats and adults were positive for the presence of Eimeria, with frequency of 62,2% and 37,8% respectively. Regarding the species found, we observed that in young Eimeria ninakohlyakimovaewas the one with a higher frequency (28.42%), unlike the adults, which highlighted the E. apsheronica(26.66%). Furthermore, also form identified as E. alijevi, E. arloingi, E. hirci, E. jolchijevi, E. christenseni, E. caprovina and E.caprina.

Parasitismo mútuo por cestóides em caprino jovem: relato de caso / Mutual parasitism by cestoid in a young goat: case report

A classe Cestoda é constituída por endoparasitas obrigatórios, cujos adultos são encontrados no intestino delgado de ruminantes causando lesões de mucosa devido à fixação de suas ventosas, além de desnutrição do animal parasitado provocada pela competição por nutrientes. O objetivo do presente estudo foi de relatar o mútuo parasitismo em um caprino jovem por Moniezia expansa em associação com outras espécies. Foram identificadas três espécies de helmintos parasitando o intestino delgado simultaneamente. Do total de 2709 exemplares adultos recuperados, 2643 eram Trichostrongylus columbriformis, um Strongyloides papillosuse 65 Moniezia expansa. Assim, registra-se, pela primeira vez, o parasitismo mútuo entre 65 M. expansa e outros helmintos.

The Cestoda class is constituted by obligatory endoparasites, whose adults are found in the small intestine of ruminants causing mucosal lesions due to fixation of their suckers, added to the malnutrition of animal caused by competition of parasites for the nutrients. The aim of this study was to report the mutual parasitism in a young goat by Moniezia expansa associated to other species. From a total of 2709 worms recovered in small intestine, 2643 were Trichostrongylus columbriformis, one Strongyloides papillosus and 65 Moniezia expansa. This way, it was registered, for the first time, the mutual parasitism by 65 M. expansa and other helminthes.

Evaluation of Eucalyptus citriodora essential oil on goat gastrointestinal nematodes.

Phytotherapy may be an alternative strategy for controlling gastrointestinal parasites. This study evaluated the anthelmintic efficacy of Eucalyptus citriodora essential oil (EcEO). The in vitro effects of EcEO were determined through testing the inhibition of egg hatching and larval development of Haemonchus contortus. EcEO was subjected to acute toxicity testing on mice, orally and intraperitoneally. The in vivo effects of EcEO were determined by the fecal egg count reduction test (FECRT) in goats infected with gastrointestinal nematodes. The results showed that 5.3 mg.mL(-1) EcEO inhibited egg hatching by 98.8% and 10.6 mg.mL(-1) EcEO inhibited H. contortus larval development by 99.71%. The lethal doses for 50% of the mice were 4153 and 622.8 mg.kg(-1), for acute toxicity orally and intraperitoneally. In the FECRT, the efficacy of EcEO and ivermectin was 66.25 and 79.16% respectively, on goat gastrointestinal nematodes eight days after treatment. EcEO showed in vitro and in vivo anthelmintic activity.