Fasciola hepatica and rumen flukes - in vitro evaluation of main commercial anthelmintics

Background: Trematode infections are of great importance as they affect the health of many species of mammals as cattle, sheep and goat. Fasciola hepatica represents the main trematode zoonosis and risks to human and cattle and paramphistomosis is one emerging parasitic diseases of ruminants widely distributed in the world. The economic expenses are incurred by the use of ineffective anthelmintics for trematode control. Besides to faecal egg count reduction test (FECRT) to determine the anthelmintic efficacy, can be used in vitro assays, by this the aim of the study was to determine the lethal doses (LD) with hatching egg test (EHT) of the main commercial anthelmintics used for the control of trematodes in cattle. Materials, Methods & Results: Liver and rumen were examined from cattle slaughtered in Tabasco, Chiapas and Campeche states from Mexico. F. hepatica eggs were recovered from gallbladder and rumen fluke eggs collected from adult parasites in saline solution. Subsequently, the hatching egg assays were performed placing 100 trematode eggs in distilled water in each one of 96 wells of polystyrene plates. After making the appropriate dilutions, several concentrations of commercial anthelmintics were evaluated, ranging from 0.04 to 80.63 mM for triclabendazole + 0.046 to 96.87 mM febendazole (TC+FBZ), from 0.04 to 91 mM for rafoxanide (RAFOX), from 0.02 to 43.74 mM for closantel (CLOS), from 0.036 to 76.18 mM for clorsulon + 0.002 to 3.31 mM ivermectin (CLORS+ IVM) and from 0.163 to 334.47 mM for nitroxynil (NITROX). A control group (water) was included in each plate. Lethal doses were obtained using the Probit procedure and analysis of the means with a one-way statistical design. Most drugs used against rumen fluke eggs presented a high LD50 and therefore were ineffective to cause egg mortality, such was the case of RAFOX that presented LD50 from 4,580 to 10,790 µg/mL (7 to 17 mM). CLOS presented the lowest LD50 (80 µg/mL or 0.12 mM) on rumen fluke eggs. TC+FBZ was found to be effective drug against the development of F. hepatica eggs in many samples. In the same way NITROX showed a low LD50 (37 to 63 µg/mL or 0.13 to 0.22 mM), but RAFOX presented a highest LD50 (1,450 µg/mL or 2.32 mM). Discussion: The present study focused on screen the ovicidal activity and determining in vitro lethal doses 50 of main commercial anthelmintics used to control F. hepatica and rumen fluke as rapid tests in a tropical region from Mexico. The FECRT is the main method to detect effectiveness of anthelmintic and other method is the coproantigen reduction test (CRT) by ELISA. Both tests require many infected animals depending the number of treatments and by this the egg hatch assay (EHA) represent a complementary diagnosis of effectiveness of anthelmintic products to compare between regions and even between farms, because few animals are required from the farm to collect trematode eggs, and it is possible to know the effectiveness against various anthelmintics at the same time. Efficacy studies on trematodes using egg hatching tests are scarce, although they have the advantage that they can be applied to both F. hepatica and rumen fluke. TC+FBZ was one of the most effective products in inhibiting the development of F. hepatica eggs. However, RAFOX showed low effectiveness against trematode eggs, with very high lethal doses. These results agree with a study that show low efficacy against the development of Paramphistomum cervi eggs and with the FECRT test reductions of 75% and 80.58% were obtained, in times from 7 to 84 days after treatment with RAFOX. NITROX and CLORS were drugs that had good efficacy on the development of F. hepatica eggs. A differential response between liver and rumen fluke was observed. The anthelmintics used against rumen fluke eggs show low ovicidal activity and in Fasciola hepatica TC+FBZ show the best activity

Effects of purge potato (operculina macrocarpa) in multinutritional blocks on sheep naturally infected with gastrointestinal helminth

Objetivo desse estudo foi avaliar os efeitos da batata de purga (O. macrocarpa) oferecida em blocos multinutricionais sobre ovinos naturalmente infectados por helmintos gastrintestinais. O trabalho foi desenvolvido de acordo com um delineamento inteiramente casualizado, com quatro tratamentos e seis repetições de um animal. Os tratamentos foram: T1 = apenas BMs; T2 = BMs + batata de purga (1g/Kg/peso vivo); T3 = BMs + batata de purga (0,5g/Kg/pv) e T4 = BMs + anti-helmíntico químico. Nos tratamentos com batata de purga 1g/Kg/pv, 0,5g/Kg/pv e químico o OPG diminuiu aos sete dias com 767, 516, 267 e eficácia de 53, 68, e 84% respectivamente. Os valores médios das hemácias, hemoglobina, hematócrito e leucócitos foram menores para o tratamento apenas com BMs, porém se mantiveram dentro dos valores de referência. Para globulina, proteínas totais, creatinina os valores médios ficaram discretamente abaixo da referência para ovinos. Conclui-se que a O. macrocarpa adicionada aos blocos multinutricionais nas doses e período estudado não provocaram respostas fisiorgânicas características de toxicidade.(AU)

This study evaluates the effects of purge potato (Operculina macrocarpa) in multi-nutritional blocks (MBs) on sheep naturally infected with gastrointestinal helminths. The experimental design was completely randomized, with four treatments and six replicates per animal. Treatments were: T1 = only MBs; T2 = MBs + purge potato (1g/Kg/lw); T3 = MBs + purge potato (0.5/Kg/lw); and T4 = MBs + chemical anthelmintic. In the treatments with 1g/Kg/lw and 0.5g/Kg/lw purge potato and with chemical anthelmintic, eggs per gram (EPG) counts decreased to 767, 516, and 267 at seven days, with an effectiveness of 53, 68, and 84%, respectively. The mean values of red blood cells, hemoglobin, hematocrit, and leukocytes were lower for the treatment with only MBs; however, they were maintained within the reference values. For globulin, total protein, and creatinine, the mean values were discretely below the reference for sheep. It is concluded that adding O. macrocarpa to the multi-nutritional blocks in the studied doses and period did not cause physiological responses characteristic of toxicity.(AU)

Anthelmintic resistance in Creole horses in the South of Rio Grande do Sul, Brazil / [Resistência anti-helmíntica em equinos da raça Crioula no sul do Rio Grande do Sul, Brasil]

The objective of the study was to evaluate the antiparasitic resistance against horse nematodes in the South of Rio Grande do Sul, Brazil. The results concerning the tests of anthelmintic efficacy on horses, stored in the database of the Parasitic Diseases Study Group (GEEP) - Veterinary Faculty, at the Federal University of Pelotas (UFPel), were carried out in the laboratory from 2018 to 2019. Stool samples were received from farms with breeding of adult female and male Criollo horses naturally infected, located in municipalities in the country's southern region. The antiparasitic agents tested were Triclorfon + Fenbendazole, Closantel + Albendazole, Ivermectin + Praziquantel, Fenbendazole, Ivermectin, Doramectin, Mebendazole and Moxidectin. Techniques such as Gordon and Whitlock, Coproculture and Fecal Egg Count Reduction Test were performed. Of all the antiparasitic drugs tested, it was observed that only treatments with Ivermectin 2% showed desired values. The observed results indicate that resistance to macrocyclic lactones is usual in equine parasites in this Brazilian region, despite the results with isolated Ivermectin.(AU)

O objetivo deste estudo é avaliar a resistência antiparasitária contra nematodeos de equinos no sul do Rio Grande do Sul, Brasil. Os resultados referentes aos testes de eficácia anti-helmíntica em cavalos, armazenados no banco de dados do Grupo de Estudos de Doenças Parasitárias (GEEP) - Faculdade de Veterinária, da Universidade Federal de Pelotas (UFPel), foram realizados em laboratório, no período de 2018 a 2019. Amostras de fezes foram recebidas de fazendas com criação de cavalos Crioulos adultos fêmeas e machos naturalmente infectados, localizadas em municípios da região Sul do país. Os agentes antiparasitários testados foram triclorfon + fenbendazol, closantel + ivermectina + praziquantel, fenbendazol, ivermectina, doramectina, mebendazol e moxidectina. Técnicas como Gordon e Whitlock, coprocultura e teste de redução da contagem de ovos fecais foram realizadas. De todos os antiparasitários testados, observou-se que apenas os tratamentos com ivermectina 2% apresentaram os valores desejados. Os resultados indicam que a resistência às lactonas macrocíclicas é comum em parasitas equinos nessa região brasileira, apesar dos resultados com ivermectina isolada.(AU)

Effects of purge potato (operculina macrocarpa) in multinutritional blocks on sheep naturally infected with gastrointestinal helminth

Objetivo desse estudo foi avaliar os efeitos da batata de purga (O. macrocarpa) oferecida em blocos multinutricionais sobre ovinos naturalmente infectados por helmintos gastrintestinais. O trabalho foi desenvolvido de acordo com um delineamento inteiramente casualizado, com quatro tratamentos e seis repetições de um animal. Os tratamentos foram: T1 = apenas BMs; T2 = BMs + batata de purga (1g/Kg/peso vivo); T3 = BMs + batata de purga (0,5g/Kg/pv) e T4 = BMs + anti-helmíntico químico. Nos tratamentos com batata de purga 1g/Kg/pv, 0,5g/Kg/pv e químico o OPG diminuiu aos sete dias com 767, 516, 267 e eficácia de 53, 68, e 84% respectivamente. Os valores médios das hemácias, hemoglobina, hematócrito e leucócitos foram menores para o tratamento apenas com BMs, porém se mantiveram dentro dos valores de referência. Para globulina, proteínas totais, creatinina os valores médios ficaram discretamente abaixo da referência para ovinos. Conclui-se que a O. macrocarpa adicionada aos blocos multinutricionais nas doses e período estudado não provocaram respostas fisiorgânicas características de toxicidade.

This study evaluates the effects of purge potato (Operculina macrocarpa) in multi-nutritional blocks (MBs) on sheep naturally infected with gastrointestinal helminths. The experimental design was completely randomized, with four treatments and six replicates per animal. Treatments were: T1 = only MBs; T2 = MBs + purge potato (1g/Kg/lw); T3 = MBs + purge potato (0.5/Kg/lw); and T4 = MBs + chemical anthelmintic. In the treatments with 1g/Kg/lw and 0.5g/Kg/lw purge potato and with chemical anthelmintic, eggs per gram (EPG) counts decreased to 767, 516, and 267 at seven days, with an effectiveness of 53, 68, and 84%, respectively. The mean values of red blood cells, hemoglobin, hematocrit, and leukocytes were lower for the treatment with only MBs; however, they were maintained within the reference values. For globulin, total protein, and creatinine, the mean values were discretely below the reference for sheep. It is concluded that adding O. macrocarpa to the multi-nutritional blocks in the studied doses and period did not cause physiological responses characteristic of toxicity.

Anthelmintic effect of four extracts obtained from Caesalpinia coriaria foliage against the eggs and larvae of Haemonchus contortus / Efeito anti-helmíntico de quatro extratos obtidos da folha de Caesalpinia coriaria contra ovos e larvas de Haemonchus contortus

To investigate the in vitro anthelmintic efficacy of dividivi (Caesalpinia coriaria), a traditional medicinal plant used in Central America and the northern part of South America, extracts from the foliage of this plant were subjected to the egg hatching test (EHT) and larval exsheathment inhibition test (LEIT), against Haemonchus contortus. Four different extracts were evaluated: acetone-water (AW), methanol-water (MW), acetone-water-dichloromethane (AWD) and methanol-water-dichloromethane (MWD). The concentrations used for the EHT and LEIT tests ranged from 500 to 4000 µg mL-1 and six repetitions per concentration. The effective concentrations (EC50) were calculated using Probit analysis. The EC50 for EHT were 2947.0, 3347.0, 3959.6 and 4538.7 µg mL-1 for MWD, MW, AW and AWD, respectively. The EC50 for LEIT were 2883.4, 5927.4, 9876.3 and 9955.4 µg mL-1 for AWD, AW, MWD and MW, respectively. The methanol extracts were the most effective in inhibiting the hatching of eggs, while the acetone extracts showed efficacy in inhibiting larval exsheathment. This study explains the importance that C. coriaria has as a medicinal plant in Central and South American countries.(AU)

Para investigar a eficácia anti-helmíntica de Divi-divi (Caesalpinia coriaria), uma planta medicinal tradicional usada na América Central e no norte da América do Sul. Extratos das folhas dessa planta foram utilizados em testes in vitro de inibição da eclosão de ovos (EHT) e desembainhamento larvar (LEIT) de Haemonchus contortus. Quatro diferentes extratos foram avaliados: acetona-água (AW), metanol-água (MW), acetona-água-diclorometano (AWD) e metanol-água-diclorometano (MWD). Para os testes EHT e LEIT, as concentrações utilizadas variaram de 500 a 4000 µg mL-1, em seis repetições por concentração. As concentrações efetivas (EC50) foram calculadas, usando-se a análise Probit. A EC50 para EHT foram 2947,0; 3347,0; 3959,6 e 4538,7 µg mL-1 para MWD, MW, AW e AWD, respectivamente. As EC50 para LEIT foram 2883,4; 5927,4; 9876,3 e 9955,4 µg mL-1 para AWD, AW, MWD e MW, respectivamente. Os extratos de metanol foram os mais eficazes em inibir a eclosão de ovos, enquanto os extratos de acetona mostraram-se eficazes em inibir a desembainhamento larvar. Este estudo ajuda a explicar a importância da C. coriaria como planta medicinal nos países da América Central e América do Sul.(AU)

Anthelmintic resistance of Haemonchus contortus from sheep flocks in Brazil: concordance of in vivo and in vitro (RESISTA-Test©) methods / Resistência anti-helmíntica de Haemonchus contortus em rebanhos ovinos noBrasil: concordância de métodos in vivo e in vitro (RESISTA-Test©)

This study evaluated the resistance status of Haemonchus contortus from sheep flocks in the state of São Paulo, Brazil, through comparison between the fecal egg count reduction test (FECRT) and the larval development test (LDT). For the FECRT, 35 sheep were selected in each of five flocks and divided into groups treated with: benzimidazole, levamisole, ivermectin, monepantel and control. Feces were collected for EPG and fecal cultures. The LDT was performed using thiabendazole (TBZ), levamisole (LEV), ivermectin aglycone (IVM-A) and Zolvix (ZLV). Resistance to all drugs was detected using FECRT in 100% of the flocks, except in relation to ZLV (40% resistant and 20% suspected of resistance). LDT indicated resistance to TBZ and IVM-A in all flocks, to LEV in 80% of flocks and to ZLV in 10%. Total agreement was obtained between the two tests for TBZ and IVM (k = 1.0), while for LEV (k = 0.8) and ZLV (k = 0.9), substantial and almost perfect agreement were obtained, respectively. The concordance between the tests was significant, thus showing that it is possible to use the outcome of the LDT to predict the FECRT, and hence validating the former as a fast diagnostic test for use by sheep farmers in Brazil.(AU)

Este estudo avaliou o status de resistência de Haemonchus contortus em rebanhos ovinos do estado de São Paulo, Brasil, através de um estudo comparativo entre o teste de redução da contagem de ovos nas fezes (TRCOF) e o teste de desenvolvimento larvar (TDL). Para o TRCOF, 35 ovinos foram selecionados em cada um dos cinco rebanhos e divididos em grupos tratados com: benzimidazol, levamisol, ivermectina, monepantel e controle. Fezes foram coletadas para OPG e coproculturas. O TDL foi realizado com tiabendazol-TBZ, levamisol-LEV, ivermectina aglicona-IVM-A e Zolvix-ZLV. Resistência a todos os fármacos foi detectada por meio do TRCOF em 100% dos rebanhos, exceto para ZLV (40% resistentes e 20% suspeitos de resistência). O LDT indicou resistência ao TBZ e IVM-A em todos os rebanhos; em 80% dos rebanhos ao LEV e 10% ao ZLV. Foi obtida concordância total entre os dois testes para TBZ e IVM (k = 1,0), enquanto para LEV (k = 0,8) e ZLV (k = 0,9) foram obtidas concordância substancial e quase perfeita, respectivamente. A concordância entre os testes foi significante, sendo possível usar o resultado do LDT para prever o do TRCOF, validando-se o primeiro como um teste diagnóstico rápido para criadores de ovinos no Brasil.(AU)

Efficacy of Tithonia diversifolia (Hemsl) A. Gray on the inhibition of larval development of Haemonchus contortus / Eficácia de Tithonia diversifolia(Hemsl) A. Gray na inibição do desenvolvimento larval de Haemonchus contortus

Sheep farming is an important economic activity from a commercial and subsistence perspective. However, helminth infections present a large sanitary problem for the production, especially haemonchosis caused by strains resistant to traditional anthelmintics. Here, we evaluated the effects of the stem and leaf of Tithonia diversifolia on the inhibition of Haemonchus contortus larval development. We performed a quantitative fecal culture test with feces of sheep inoculated with larvae tolerant to albendazole. Fecal cultures were treated with leaf or stem powder and were compared to the controls containing distilled water or levamisole. The T. diversifolia leaf and stem powders significantly reduced the production of H. contortus larvae. However, leaf powder was more effective, with 64.95% efficacy in the reduction of larvae production, representing a promising alternative in the integrated control of this nematode.

A ovinocultura é uma importante atividade econômica em escala comercial e de subsistência. No entanto, um dos maiores desafios nessa atividade são as verminoses, destacando-se a haemoncose ocasionada por cepas resistentes a anti-helmínticos. Neste estudo objetivou-se avaliar o efeito do caule e folha de Tithonia diversifolia soba inibição do desenvolvimento de larvas de Haemonchus contortus. Foram realizadas coproculturas quantitativas com fezes de ovinos inoculados com larvas do nematódeo toleranteao albendazol. Foram avaliadas as produções de larvas infectantes em coproculturas tratadas: com pó da folha e do caule e com o extrato aquoso do caule da planta comparando-se com o tratamento controle contendo água destilada e levamisol. O pó da folha e do caule de T. diversifoliareduziram significativamente a produção de larvas infectantes de H. contortus. Entretanto, o pó das folhas foi mais efetivo, promovendo eficácia de 64,95% na redução de larvas em coproculturas, representando uma alternativa promissora no controle integrado do nematódeo.

Multiple resistance in equine cyathostomins: a case study from military establishments in Rio Grande do Sul, Brazil / Múltipla resistência de ciatostomíneos de equinos: um estudo de caso em estabelecimentos militares no Rio Grande do Sul, Brasil

Semi-intensive equine breeding system favors gastrointestinal nematode infections. The treatment of these infections is based on the use of anthelmintics. However, the inappropriate use of these drugs has led to parasitic resistance to the available active principles. The objective of this study was to evaluate the efficacy of the main classes of antiparasitic (ATP) used in control in adult and young animals, including: benzimidazoles (fenbendazole), pyrimidines (pyrantel pamoate), macrocyclic lactones (ivermectin and moxidectin), as well as the combination of active ingredients (ivermectin + pyrantel pamoate). The study was carried out in two military establishments, located in Rio Grande do Sul (RS), from January to December, 2018. The intervals between the treatments of the animals were performed from 30 to 90 days. Coproparasitological evaluations were determined by the egg count reduction in the faeces. Cyatostomine larvae were identified in pre and post-treatment cultures. The results demonstrated the multiple parasitic resistance of cyathostomins to fenbendazole, moxidectin in young animals, and to fenbendazole, pyrantel pamoate in adult animals. Thus, it is necessary to define or diagnose parasitic resistance to assist in the creation of prophylactic parasitic control, using suppressive treatment with ATP associated with integrated alternatives. The progress of parasitic resistance can be slowed.(AU)

O sistema semi-intensivo de criação de equinos favorece infecções por nematoides gastrointestinais. O tratamento dessas infecções é baseado no uso de anti-helmínticos. No entanto, o uso inadequado desses medicamentos levou à resistência parasitária aos princípios ativos disponíveis. O objetivo deste estudo foi avaliar a eficácia das principais classes de antiparasitários (ATP), utilizados no controle em animais adultos e jovens, incluindo: benzimidazois (fenbendazol), pirimidinas (pamoato de pirantel), lactonas macrocíclicas (ivermectina e moxidectina), bem como a combinação de ingredientes ativos (ivermectina + pamoato de pirantel). O estudo foi realizado em dois estabelecimentos militares, localizados no Rio Grande do Sul (RS), de janeiro a dezembro de 2018. Os intervalos entre os tratamentos foram realizados de 30 a 90 dias. As avaliações coproparasitológicas foram determinadas pela redução da contagem de ovos nas fezes. Foram identificadas larvas de ciatostomíneos nas coproculturas pré e pós tratamentos. Os resultados demonstraram a resistência parasitária múltipla dos ciatostomíneos ao fenbendazol, moxidectina em animais jovens, febendazole e pamoato de pirantel em animais adultos. Estabelecer o diagnóstico da resistência parasitária auxiliará na elaboração de um controle parasitário profilático, reduzindo o tratamento supressivo com ATP juntamente com alternativas de controle integrado. Dessa forma, o avanço da resistência parasitária poderá ser retardado.(AU)

Efficacy of Tithonia diversifolia (Hemsl) A. Gray on the inhibition of larval development of Haemonchus contortus / Eficácia de Tithonia diversifolia(Hemsl) A. Gray na inibição do desenvolvimento larval de Haemonchus contortus

Sheep farming is an important economic activity from a commercial and subsistence perspective. However, helminth infections present a large sanitary problem for the production, especially haemonchosis caused by strains resistant to traditional anthelmintics. Here, we evaluated the effects of the stem and leaf of Tithonia diversifolia on the inhibition of Haemonchus contortus larval development. We performed a quantitative fecal culture test with feces of sheep inoculated with larvae tolerant to albendazole. Fecal cultures were treated with leaf or stem powder and were compared to the controls containing distilled water or levamisole. The T. diversifolia leaf and stem powders significantly reduced the production of H. contortus larvae. However, leaf powder was more effective, with 64.95% efficacy in the reduction of larvae production, representing a promising alternative in the integrated control of this nematode.  

A ovinocultura é uma importante atividade econômica em escala comercial e de subsistência. No entanto, um dos maiores desafios nessa atividade são as verminoses, destacando-se a haemoncose ocasionada por cepas resistentes a anti-helmínticos. Neste estudo objetivou-se avaliar o efeito do caule e folha de Tithonia diversifolia soba inibição do desenvolvimento de larvas de Haemonchus contortus. Foram realizadas coproculturas quantitativas com fezes de ovinos inoculados com larvas do nematódeo toleranteao albendazol. Foram avaliadas as produções de larvas infectantes em coproculturas tratadas: com pó da folha e do caule e com o extrato aquoso do caule da planta comparando-se com o tratamento controle contendo água destilada e levamisol. O pó da folha e do caule de T. diversifoliareduziram significativamente a produção de larvas infectantes de H. contortus. Entretanto, o pó das folhas foi mais efetivo, promovendo eficácia de 64,95% na redução de larvas em coproculturas, representando uma alternativa promissora no controle integrado do nematódeo.(AU)

Effect of nutraceuticals on acanthocephalan Neoechinorhynchus buttnerae and its toxicity to the host tambaqui Colossoma macropomum.

The production of tambaqui Colossoma macropomum has been undergoing financial losses due to parasitic infection by the acanthocephalan Neoechinorhynchus buttnerae, raising an alert for aquaculture in South America. The lack of adequate treatment and use of unlicensed chemicals encourages research for alternative solutions with minimal side effects. The objectives of this study were to evaluate the in vitro antiparasitic potential of commercial nutraceutical products (Natumix® and BioFish®) against N. buttnerae and to assess the respective in vivo toxic effects on the host tambaqui. For in vitro assays, parasitized fish were necropsied for acanthocephalans sampling. The parasites were exposed to three concentrations (0.078, 0.313 and 1.25 mg/ml) of each product, as well as controls (one without product and another with a solubilizer). For the in vivo acute toxicity test, juvenile fish (<0.1 g) were exposed to five increasing concentrations of each product. Mortality of tambaqui was recorded at 24, 48, 72 and 96 h. The estimated lethal concentration (LC) for 10, 50, 90 and 99% of fish was determined to classify the toxicity of the products on the target species. After in vitro efficacy tests, the highest concentrations (1.25 mg/ml) caused 100% mortality of the parasites in both products, but only Natumix® caused 100% mortality using the intermediate concentration (0.313 mg/ml) after 24 h. According to the acute toxicity result, the LC50 classified the nutraceutical products as slightly toxic for tambaqui. The tested products had a parasiticidal effect on N. buttnerae, and the toxicity test showed that both products have therapeutic potential when added to the diet.

Araticum (Annona crassiflora Mart.) as a source of nutrients and bioactive compounds for food and non-food purposes: A comprehensive review.

Araticum (Annona crassiflora Mart.) is a fruitful tree native to the Brazilian Cerrado biome that holds high nutritional, functional and economic potential. This plant has been used since ancient times by folk medicine for the treatment of several pathological conditions. There has been increasing interest in the development of pulp-based food products as well as the by-products utilization to obtain value-added ingredients. Understanding the chemical composition and biological activities of different botanical parts of Annona crassiflora Mart. provides a basis to support future researches and applications. In this context, this paper carries out an exhaustive review of the scientific literature, on the main phytochemicals of different botanical parts of Annona crassiflora Mart. (fruit, leaves, stem and root) and their biological activities, assessing their potential uses for several industrial segments. Annona crassiflora Mart. fruits and especially their by-products (peel and seeds) and leaves have been shown a wide range of bioactive compounds such as phenolic compounds, alkaloids, annonaceous acetogenins, tocols, carotenoids, phytosterols, dietary fiber, vitamins, minerals and essential oils. These compounds contribute to various biological activities, including antioxidant, hepatoprotective, anti-inflammatory, antitumoral, analgesic, antidiabetic, skin healing, antidiarrhoeic, antimicrobial, antiparasitic, insecticide and herbicide activities of Annona crassiflora Mart. extracts. Therefore, these findings demonstrate that Annona crassiflora Mart. fruit, by-products and leaves can be excellent candidates to be used as functional foods and/or sources for obtaining bioactive compounds for the food, cosmetics and pharmaceutical applications.

Efficacy, toxicity, and lethality of plants with potential anthelmintic activity in small ruminants in Brazil / Eficácia, toxicidade e letalidade de plantas com potencial atividade anti-helmíntica em pequenos ruminantes no Brasil

The use of medicinal plants as a therapeutic method in the control of diseases has been increasing in animal production. In the case of small ruminants, the endoparasitic disease is a major concern, since they are considered the greater sanitary problem, especially when considering the phenomenon of parasite resistance to the medicines used. Therefore, the development of alternative methods of endoparasitary control in goats and sheep has become a pressing need. The use of plants to control endoparasites can reduce the use of chemical inputs, making more environmentally sustainable livestock, minimizing the occurrence of parasitic resistance to conventional medicines and contributing to the reduction of production costs. In this sense, several studies have been carried out in order to evaluate the in vitro and in vivo activity of plants with anthelmintic potential. However, despite the potentiality found in many studies, the results are sometimes contradictory or do not replicate in vivo the same efficacy obtained in the in vitro assays. Another relevant aspect is the low utilization of the lethality and toxicity tests, which are indispensable so that the knowledge can be spread and applied by the producers in their herds. Thus, this review aims to provide the results from studies carried out in Brazil with extract of plants with potential for control of parasitic disease in goats and sheep, describing the main evaluated plants species, mechanisms of action, preparation forms and tests of efficacy, toxicity, and lethality.(AU)

A utilização de plantas medicinais como método terapêutico no controle de enfermidades tem sido crescente na produção animal. No caso de pequenos ruminantes, as verminoses representam uma grande preocupação, visto que são consideradas o maior problema sanitário, especialmente quando se considera o fenômeno da resistência dos parasitas aos fármacos utilizados. Portanto, o desenvolvimento de métodos alternativos de controle endoparasitário em caprinos e ovinos tem se tornado uma necessidade premente. O uso de plantas para controlar as endoparasitoses pode reduzir o uso de insumos químicos, tornando a atividade pecuária mais ambientalmente sustentável, minimizando a ocorrência da resistência parasitária aos medicamentos convencionais e contribuindo para redução dos custos de produção. Nesse sentido, diversos estudos têm sido executados a fim de se avaliar a atividade in vitro e in vivo de plantas com potencial anti-helmíntico. Contudo, a despeito da potencialidade verificada em muitos estudos, os resultados são, por vezes, contraditórios ou não reproduzem in vivo a mesma eficácia obtida nos ensaios in vitro. Outro aspecto relevante é a baixa utilização dos testes de letalidade e toxicidade, indispensáveis para que o conhecimento possa ser difundido e aplicado pelos produtores rurais. Assim, essa revisão visa fornecer os resultados obtidos nos estudos realizados no Brasil com extratos de plantas com potencial para controle de verminose em caprinos e ovinos, descrevendo as principais espécies vegetais avaliadas, mecanismos de ação, formas de preparo e os testes de eficácia, toxicidade e letalidade.(AU)

Efficacy, toxicity, and lethality of plants with potential anthelmintic activity in small ruminants in Brazil / Eficácia, toxicidade e letalidade de plantas com potencial atividade anti-helmíntica em pequenos ruminantes no Brasil

The use of medicinal plants as a therapeutic method in the control of diseases has been increasing in animal production. In the case of small ruminants, the endoparasitic disease is a major concern, since they are considered the greater sanitary problem, especially when considering the phenomenon of parasite resistance to the medicines used. Therefore, the development of alternative methods of endoparasitary control in goats and sheep has become a pressing need. The use of plants to control endoparasites can reduce the use of chemical inputs, making more environmentally sustainable livestock, minimizing the occurrence of parasitic resistance to conventional medicines and contributing to the reduction of production costs. In this sense, several studies have been carried out in order to evaluate the in vitro and in vivo activity of plants with anthelmintic potential. However, despite the potentiality found in many studies, the results are sometimes contradictory or do not replicate in vivo the same efficacy obtained in the in vitro assays. Another relevant aspect is the low utilization of the lethality and toxicity tests, which are indispensable so that the knowledge can be spread and applied by the producers in their herds. Thus, this review aims to provide the results from studies carried out in Brazil with extract of plants with potential for control of parasitic disease in goats and sheep, describing the main evaluated plants species, mechanisms of action, preparation forms and tests of efficacy, toxicity, and lethality.

A utilização de plantas medicinais como método terapêutico no controle de enfermidades tem sido crescente na produção animal. No caso de pequenos ruminantes, as verminoses representam uma grande preocupação, visto que são consideradas o maior problema sanitário, especialmente quando se considera o fenômeno da resistência dos parasitas aos fármacos utilizados. Portanto, o desenvolvimento de métodos alternativos de controle endoparasitário em caprinos e ovinos tem se tornado uma necessidade premente. O uso de plantas para controlar as endoparasitoses pode reduzir o uso de insumos químicos, tornando a atividade pecuária mais ambientalmente sustentável, minimizando a ocorrência da resistência parasitária aos medicamentos convencionais e contribuindo para redução dos custos de produção. Nesse sentido, diversos estudos têm sido executados a fim de se avaliar a atividade in vitro e in vivo de plantas com potencial anti-helmíntico. Contudo, a despeito da potencialidade verificada em muitos estudos, os resultados são, por vezes, contraditórios ou não reproduzem in vivo a mesma eficácia obtida nos ensaios in vitro. Outro aspecto relevante é a baixa utilização dos testes de letalidade e toxicidade, indispensáveis para que o conhecimento possa ser difundido e aplicado pelos produtores rurais. Assim, essa revisão visa fornecer os resultados obtidos nos estudos realizados no Brasil com extratos de plantas com potencial para controle de verminose em caprinos e ovinos, descrevendo as principais espécies vegetais avaliadas, mecanismos de ação, formas de preparo e os testes de eficácia, toxicidade e letalidade.

Assessment of the long-acting ivermectin formulation in sheep: Further insight into potential pharmacokinetic interactions.

The aim of the current study was to evaluate the in vivo pharmacokinetic of ivermectin (IVM) after the administration of a long-acting (LA) formulation to sheep and its impact on potential drug-drug interactions. The work included the evaluation of the comparative plasma profiles of IVM administered at a single therapeutic dose (200 µg/kg) and as LA formulation at 630 µg/kg. Additionally, IVM was measured in different gastrointestinal tissues at 15 days posttreatment with both IVM formulations. The impact of the long-lasting and enhanced IVM exposure on the disposition kinetics of abamectin (ABM) was also assessed. Plasma (IVM and ABM) and gastrointestinal (IVM) concentrations were analyzed by HPLC with fluorescent detection. In plasma, the calculated Cmax and AUC0-t values of the IVM-LA formulation were 1.47- and 3.35-fold higher compared with IVM 1% formulation, respectively. The T1/2ab and Tmax collected after administration of the LA formulation were 2- and 3.5-fold longer than those observed after administration of IVM 1% formulation, respectively. Significantly higher IVM concentrations were measured in the intestine mucosal tissues and luminal contents with the LA formulation, and in the liver, the increase was 7-fold higher than conventional formulation. There was no drug interaction between IVM and ABM after the single administration of ABM at 15 days post-administration of the IVM LA formulation. The characterization of the kinetic behavior of the LA formulation to sheep and its potential influence on drug-drug interactions is a further contribution to the field.

Anthelmintic activity of Eucalyptus citriodora essential oil and its major component, citronellal, on sheep gastrointestinal nematodes / Atividade anti-helmíntica do óleo essencial de Eucalyptus citriodora e seu componente majoritário, citronelal, sobre nematoides gastrintestinais de ovinos

This study aimed to evaluate the anthelmintic activity of Eucalyptus citriodora essential oil and citronellal on sheep gastrointestinal nematodes. Essential oil composition was determined by gas chromatography mass spectrometry. The substances were evaluated in vitro using adult worm motility test (AWMT) and transmission electron microscopy (TEM). The acute toxicity test in mice and the fecal egg count reduction test (FECRT) in sheep were performed. Citronellal was confirmed as the essential oil major constituent (63.9%). According to the AWMT, 2 mg/mL of essential oil and citronellal completely inhibited Haemonchus contortus motility at 6 h post exposure. H. contortus exposed to essential oil and citronellal exhibited internal ultrastructural modifications. The lethal dose 50 values in mice were 5,000 and 2,609 mg/kg for essential oil and citronellal, respectively. E. citriodora essential oil reduced sheep epg at 14 days post treatment by 69.5% (P 0.05). No significant differences were observed in epg between the citronellal and negative control groups (P>0.05). The interaction between citronellal and other constituents in the essential oil may be relevant for its in vivo anthelmintic activity. Thus, E. citriodora essential oil and citronellal pharmacokinetic studies may help elucidate the anthelmintic activity of these compounds.(AU)

Este trabalho objetivou avaliar a atividade anti-helmíntica do óleo essencial de Eucalyptus citriodora e citronelal sobre nematoides gastrintestinais de ovinos. A composição do óleo essencial foi determinada por cromatografia gasosa acoplada à espectrometria de massas. As substâncias foram avaliadas in vitro utilizando-se teste de motilidade de vermes adultos (AWMT) e microscopia eletrônica de transmissão (TEM). Teste de toxicidade aguda em camundongos e teste de redução da contagem de ovos fecais (FECRT) em ovinos foram realizados. Citronelal foi confirmado como componente majoritário do óleo essencial (63,9%). No AWMT, 2 mg/mL de óleo essencial e citronelal inibiram completamente a motilidade de H. contortus 6 h pós-exposição. H. contortus expostos ao óleo essencial e citronelal exibiram modificações ultraestruturais internas. Os valores da dose letal 50 em camundongos foram 5.000 e 2.609 mg/kg para óleo essencial e citronelal, respectivamente. Óleo essencial de E. citriodora reduziu opg de ovinos 14 dias pós-tratamento em 69,5% (P 0,05). Não houve diferença significativa de opg entre grupo controle negativo e citronelal (P>0,05). A interação entre citronelal e outros constituintes do óleo essencial pode ser relevante na atividade anti-helmíntica in vivo. Portanto, avaliação farmacocinética do óleo essencial de E. citriodora e citronelal pode auxiliar a elucidar a atividade anti-helmíntica desses compostos.(AU)

Implication of the fecal egg count reduction test (FECRT) in sheep for better use of available drugs / Implicação do teste de redução da contagem de ovos nas fezes (TRCOF) em ovinos para melhor uso dos anti-helmínticos disponíveis

The aim here is to present data on the efficacy of anthelmintics in sheep flocks in Rio de Janeiro, Brazil, and to discuss the interpretation of the fecal egg count reduction test (FECRT) for each nematode genus. Fecal eggs counts and pre- and post-treatment coprocultures were performed, the former to evaluate the efficacy of and the latter to determine the overall parasite prevalence. An additional efficacy test was performed at Farm / 1 a year after the initial test. Severe anthelmintic resistance was found for the flocks, with no FECRT sensitivity at any of the 22 farms evaluated. However, an analysis of the infective larvae showed that some drugs were effective against certain parasitic genera; i.e., levamisole was more effective against Haemonchus spp. and moxidectin against Trichostrongylus spp. In the additional FECRT performed at Farm / 1, moxidectin and nitroxynil were ineffective separately, but when applied in combination they were highly effective due to their efficacy against Haemonchus (nitroxynil) and Trichostrongylus (moxidectin), respectively. The use of the FECRT targeting the parasitic nematode species prevalent on farms may make it possible to choose more effective anthelmintics.(AU)

O objetivo deste trabalho foi apresentar dados sobre a eficácia de anti-helmínticos em rebanhos ovinos no Rio de Janeiro, Brasil, e discutir a interpretação do teste de redução da contagem de ovos nas fezes (TRCOF) para cada gênero de nematoide. A contagem de ovos fecais (OPG) e coprocultura pré e pós-tratamento foram realizadas para avaliar a eficácia e a prevalência geral do parasito, respectivamente. Um teste de eficácia adicional foi realizado na Fazenda / 1 após um ano do teste inicial. Resistência anti-helmíntica grave foi encontrada, não havendo sensibilidade no TRCOF em nenhuma das 22 fazendas avaliadas. No entanto, na análise das larvas infectantes observou-se que algumas drogas foram eficazes contra certos gêneros parasitários; por exemplo, o levamisol foi mais eficaz contra Haemonchus spp. e a moxidectina contra Trichostrongylus spp. No TRCOF adicional realizado na Fazenda 1, a moxidectina e o nitroxinil foram ineficazes separadamente, mas quando aplicados em combinação, foram altamente eficazes devido à sua eficácia contra Haemonchus spp. (nitroxinil) e Trichostrongylus spp. (moxidectina), respectivamente. O TRCOF visando às espécies de nematoides parasitas prevalentes nas fazendas pode possibilitar a escolha de anti-helmínticos mais eficazes.(AU)

Anthelmintic effect of Cymbopogon citratus essential oil and its nanoemulsion on sheep gastrointestinal nematodes / Efeito anti-helmíntico do óleo essencial de Cymbopogon citratus e sua nanoemulsão sobre nematoides gastrintestinais de ovinos

The anthelmintic resistance stimulated the search for strategies for controlling gastrointestinal nematodes, including the use of free essential oils or its nanoemulsion. This study evaluated the anthelmintic efficacy of Cymbopogon citratus essential oil (CcEO) and C. citratus essential oil nanoemulsion (CcEOn). Pysicochemical analyses were performed. The in vitro effect was determined using the egg hatch test (EHT) on Haemonchus contortus and in vivo effect was evaluated in sheep infected with gastrointestinal nematodes. The animals were treated with CcEO (500 mg/kg) or CcEOn (450 mg/kg) for the fecal egg count (FEC) and the determination of worm burden. The main component of CcEO was citral. The CcEO content in the nanoemulsion was 20% (v/v), and the mean particle size was 248 nm. In EHT, CcEO and CcEOn (1.25 mg/mL) inhibited larval hatching by 98.4 and 97.1%, respectively. Three animals treated with CcEO died whereas in the group treated with CcEOn one animal died. The FEC and total worm burden of the treated groups did not differ from the negative control (p>0.05). The CcEOn showed efficacy only on H. contortus (p 0.05). In conclusion, nanoencapsulation reduced toxicity and increased efficacy on H. contortus.(AU)

A resistência anti-helmíntica estimulou a busca por estratégias de controle de nematoides gastrintestinais, incluindo óleos essenciais livres ou em nanoemulsão. Este estudo avaliou a eficácia anti-helmíntica do óleo essencial de Cymbopogon citratus (OECc) e da nanoemulsão do óleo essencial de C. citratus (nOECc). Análises físico-químicas foram realizadas. O efeito in vitro foi determinado no teste de eclosão de ovos (TEO) sobre Haemonchus contortus e o efeito in vivo foi avaliado em ovinos infectados com nematoides gastrintestinais. Os animais foram tratados com OECc (500 mg/kg) ou nOECc (450 mg/kg) para determinação do número de ovos por grama de fezes (OPG) e carga parasitária. O principal constituinte do OECc foi citral. O conteúdo de OECc na nanoemulsão foi 20% e o tamanho médio de partícula foi 248 nm. No TEO, OECc e nOECc (1,25 mg/mL) inibiram 98,4 e 97,1% da eclosão larvar, respectivamente. Três animais tratados com o OECc morreram, enquanto um animal do grupo tratado com a nOECc morreu. O OPG e a carga parasitária total dos grupos tratados não diferiram do controle negativo (p>0.05). A nOECc apresentou eficácia somente sobre H. contortus (p 0.05). Como conclusão, o nanoencapsulamento reduziu a toxicidade e aumentou a eficácia sobre H. contortus.(AU)

A cysteine protease from the latex of Ficus benjamina has in vitro anthelmintic activity against Haemonchus contortus / Atividade anti-helmíntica in vitro de uma protease cisteínica do látex de Ficus benjamina contra Haemonchus contortus

Haemonchus contortus is a gastrointestinal nematode that is responsible for high mortality rates in ruminant herds. The resistance of nematodes to synthetic anthelmintics is widespread and requires a continuous search for new bioactive molecules, such as proteins. The objective of this study was to evaluate the anthelmintic potential of a protease purified from the latex of Ficus benjamina against H. contortus . Fresh latex was collected from plants via small incisions in the green stems, the rubber was removed by centrifugation, and the latex protein extract (LPE) was obtained. After LPE fractionation with ammonium sulfate and chromatography of the fraction containing the highest proteolytic activity on CM-cellulose, a cysteine protease (FbP) was purified. FbP has a molecular mass of approximately 23.97 kDa, and its proteolytic activity was stable between pH 6.0 and pH 10 and over a broad temperature range, with optimum activity at 60 °C. FbP inhibited both the development and exsheathment of H. contortus larvae, with 50% effective concentrations of 0.26 and 0.79 mg/mL, respectively. We conclude that this cysteine protease from F. benjamina latex with anthelmintic activity against H. contortus could be a promising alternative for the development of products for use in parasite control programmes.(AU)

Haemonchus contortus é um nematoide gastrintestinal, responsável por altas taxas de mortalidade em rebanhos de pequenos ruminantes. A resistência dos nematoides aos anti-helmínticos sintéticos está generalizada e requer uma busca contínua por novos compostos bioativos, como as proteínas. O objetivo deste trabalho foi avaliar o potencial anti-helmíntico da protease purificada do látex de Ficus benjamina contra H. contortus . O látex fresco foi coletado das plantas por pequenas incisões nas hastes verdes e o extrato proteico de látex (EPL) foi obtido. Após o fracionamento do EPL com sulfato de amônio e cromatografia da fração contendo a maior atividade proteolítica da CM-Celulose, uma protease cisteínica (FbP) foi purificada. A FbP tem massa molecular de cerca de 23,97 kDa, a atividade proteolítica foi estável entre pH 6,0 e pH 10 e ao longo de uma ampla faixa de temperatura, com atividade ótima a 60 °C. A FbP inibiu tanto o desenvolvimento quanto o desembainhamento das larvas de H. contortus, com 50% de inibição nas concentrações de 0,26 e 0,79 mg/mL, respectivamente. Concluímos que esta protease cisteínica do látex de F. benjamina, com ação anti-helmíntica contra H. contortus, pode ser uma alternativa promissora para o desenvolvimento de produtos a serem utilizados em programas de controle de parasitos.(AU)

Nematode resistance to five anthelmintic classes in naturally infected sheep herds in Northeastern Brazil / Resistência de nematódeos a cinco classes de anti-helmínticos em rebanhos de ovinos naturalmente infectados no Nordeste do Brasil

This study aimed to evaluate the occurrence of nematode resistance to anthelmintics on sheep herds from the semi-arid region of Paraíba, Northeastern Brazil. Fecal Egg Count Reduction Test (FECRT) was carried out on 20 properties. In each herd, 30 animals were divided into five groups containing six animals each: group I, treated with albendazole 10%, 4 mg/kg; group II, ivermectin 0.08%, 0.2 mg/kg; group III, closantel 10%, 10 mg/kg; group IV, levamisole hydrochloride 5%, 5 mg/kg; and group V, monepantel 2.5%, 2.5 mg/kg. All treatments were administered orally as a single dose. Fecal samples were collected on days zero and 10 after treatment, to perform FECRT and coprocultures. Multiresistance was observed in all the properties; 95% had high resistance to albendazole, 85% to ivermectin, 80% to closantel, 40% to levamisole, and 45% to monepantel. On property 15, where monepantel was ineffective, a second FECRT for this anthelmintic was carried out 4 months after the first, resulting in 75% efficacy. Immediately after the FECRT result, two ewes were euthanized and necropsied, and Haemonchus contortus, Trichostrongylus axei, Trichostrongylus colubriformis, Oesophagostomum columbianum, and Trichuris ovis were recovered. It was concluded that the resistance of sheep gastrointestinal nematodes to antthelmintic, including monepantel, is high.(AU)

O objetivo deste estudo foi avaliar a ocorrência de resistência de nematódeos a anti-helmínticos em rebanhos ovinos do semiárido da Paraíba, Nordeste do Brasil. Testes de Redução na Contagem de Ovos Fecais (TRCOF) foram conduzidos em 20 propriedades. Em cada rebanho, 30 animais foram selecionados e foram divididos em cinco grupos contendo seis animais cada: grupo I, tratado com albendazol 10%, 10 mg/kg; grupo II, ivermectina 0,08%, 0,2 mg/kg; grupo III, closantel 10%, 10 mg/kg; grupo IV, cloridrato de levamisol 5%, 5 mg/kg; grupo V, monepantel 2,5%, 2,5 mg/kg. Todos os tratamentos foram administrados via oral, em dose única. Amostras fecais foram coletadas nos dias zero e 10 após os tratamentos, para a realização do TRCOF e coproculturas. Multirresistência foi observada em todas as propriedades; 95% dos rebanhos obtiveram alta resistência ao albendazole, 85% a ivermectina, 80% ao closantel, 40% ao levamisole, e 45% a monepantel. Na propriedade 15, onde o monepantel foi ineficaz, um segundo TRCOF foi conduzido para este fármaco 4 meses após o primeiro, resultando em 75% de eficácia. Imediatamente após o resultado do TRCOF, duas ovelhas foram eutanaziadas e necropsiadas, e Haemonchus contortus, Trichostrongylus axei, Trichostrongylus colubriformis, Oesophagostomum columbianum, e Trichuris ovis foram recuperados. Concluiu-se que a resistência dos nematódeos gastrintestinais de ovinos aos anti-helmínticos, incluindo o monepantel, é alta.(AU)

Albendazole treatment in laying hens: Egg residues and its effects on fertility and hatchability.

This work characterized the egg residual concentrations of albendazole (ABZ) and its sulphoxide (ABZSO) and sulphone (ABZSO2 ) metabolites and evaluated their effect on egg fertility and hatchability after ABZ treatments to laying hens. Seventy hens were allocated in groups: Group-1 was the control without treatment; Group-2 received a single ABZ oral dose (10 mg/kg); Group-3, -4 and -5 were treated with ABZ in medicated feed over 7 days at 10, 40, or 80 mg kg-1  day-1 , respectively. Eggs were analyzed to determine the ABZ/metabolite level by HPLC or subjected to incubation to evaluate the fertility and hatchability. Only ABZSO and ABZSO2 metabolites were quantified in egg after ABZ single oral administration with maximum concentrations of 0.47 ± 0.08 and 0.30 ± 0.07 µg/ml, respectively. ABZ and its metabolites were found in eggs after 7-day ABZ treatments. The egg residue exposure estimated as AUCs (areas under the concentration vs. time curve) were 100.5 (ABZ), 56.3 (ABZSO) and 141.3 µg hr g-1 (ABZSO2 ). ABZ administration did not affect the egg fertility at any dosages. Egg hatchability was not affected by ABZ treatment at 10 mg/kg in medicated feed, but it decreased when the dose was 4-8 times higher. These results should be considered when ABZ is used for deworming laying hens.