Anthelmintic resistance in cattle nematodes in northwestern São Paulo state, Brazil

The present study was carried out in the northwestern region of São Paulo State, Brazil, to determine the anthelmintic resistance status in cattle naturally infected with gastrointestinal nematodes. The anthelmintics tested were levamisole phosphate (Ripercol (R), Fort Dodge), albendazole sulphoxide (Ricobendazole, Fort Dodge), ivermectin (Ivomec (R), Merial) and moxidectin (Cydectin (R), Fort Dodge), administered at the doses recommended by the manufacturers. From April 2002 to May 2004, 25 cattle farms were evaluated. on each farm, steers were divided into treatment and control (not treated) groups based on fecal egg counts (FEC). Between 7 and 10 days after the anthelmintics administration, fecal samples were collected from each animal for post-treatment FEC. Fecal cultures from each group were also prepared for larval identification. After treatment, mean FEC reduction (FECR) in treatment groups (compared with control groups) was assessed on each farm. FECR was lower than 90% on 23 farms after ivermectin treatment. on 19 farms, FECR of 100% was recorded following moxidectin treatment; on the remaining 6, FECR ranged from 90% to 97.2%. After albendazole treatment, FECR was higher than 90% on 20 farms and ranged from 47.4% to 84.6% on other 5. After levamisole treatment, FECR was higher than 90% on 23 farms and equal to 47.4% and 73.7% on other 2 farms. Results indicated the presence of resistant Cooperia spp. and Haemonchus spp., especially to ivermectin; on some farms, resistance to albendazole and levamisole was also observed. (C) 2007 Elsevier B.V. All rights reserved.

Evaluation of a larval development assay for the detection of anthelmintic resistance in Ostertagia circumcincta

Two experiments were carried out to evaluate a larval development assay for the detection of anthelmintic resistance in O. circumcincta. In Experiment I, the dose responses to levamisole (LEV), thiabendazole (TBZ) and ivermectin (IVM) of 8 isolates of O. circumcincta were measured 34 days after infection (DAI). Four of these isolates were shown to be resistant to 1 or more anthelmintics. With 2 exceptions, all isolates considered to be resistant had higher LD50 values than the susceptible isolates for that anthelmintic. One exception was isolate RM8, which was considered to be resistant to all 3 anthelmintics based on faecal egg count reduction tests in goats, but the LD50 value for LEV did not differ from that for the susceptible isolates. The other exception was an isolate considered to be susceptible to TBZ which had a relatively high LD50 value. In an unrelated trial that was prompted by this finding, this isolate was confirmed to be benzimidazole-resistant. Isolate RM8 and an isolate susceptible to all 3 anthelmintics (SK2) were used in the second experiment, which was conducted to monitor changes in the LD50 values of LEV, TBZ and IVM over time following a single infection of 35 000 infective larvae in young sheep. Faecal samples were collected weekly from 24 to 115 DAI. With all 3 anthelmintics, the LD50 values increased with time to a peak around 50-60 DAI, and then declined to levels similar to those observed soon after patency. This trend was consistent for both isolates. The highest mean LD50 values for isolates SK2 for IVM and TBZ and RM8 for IVM and RM8, respectively, were 1.7 and 1.8 times, and 2.2 and 2.9 times higher than the initial mean LD50 values. There was a clear distinction in LD50 values between isolates at each sampling day for both IVM and TBZ. However, as a consequence of the changes in LD50 values with time, the peak LD50 values of IVM for isolate SK2 were higher than the minimum LD50 values of isolate RM8. As there was no apparent difference in LEV efficacy between these 2 isolates, the data were pooled. The highest mean LD50 value was 2.3 times higher than the initial LD50 value. (C) 1997 Australian Society for Parasitology.

Diagnosis of anthelmintic resistance in cattle in Brazil: a comparison of different methodologies

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Anthelmintic activity of Cymbopogon martinii, Cymbopogon schoenanthus and Mentha piperita essential oils evaluated in four different in vitro tests

Anthelmintic resistance is a worldwide concern in small ruminant industry and new plant-derived compounds are being studied for their potential use against gastrointestinal nematodes. Mentha piperita, Cymbopogon martinii and Cymbopogon schoenanthus essential oils were evaluated against developmental stages of trichostrongylids from sheep naturally infected (95% Haemonchus contortus and 5% Trichostrogylus spp.) through the egg hatch assay (EHA), larval development assay (LDA), larval feeding inhibition assay (LFIA), and the larval exsheathment assay (LEA). The major constituent of the essential oils, quantified by gas chromatography for M. piperita oil was menthol (42.5%), while for C. martinii and C. schoenanthus the main component was geraniol (81.4% and 62.5%, respectively). In all in vitro tests C. schoenanthus essential oil had the best activity against ovine trichostrongylids followed by C. martini, while M. piperita presented the least activity. Cymbopogon schoenanthus essential oil had LC(50) value of 0.045 mg/ml in EHA, 0.063 mg/ml in LDA, 0.009 mg/ml in LFIA, and 24.66 mg/ml in LEA. The anthelmintic activity of essential oils followed the same pattern in all in vitro tests, suggesting C. schoenanthus essential oil could be an interesting candidate for nematode control, although in vivo studies are necessary to validate the anthelmintic properties of this oil. (C) 2011 Elsevier B.V. All rights reserved.

Why is it important to correctly identify Haemonchus species?

Infecções por Haemonchus spp. são uma das principais causas de perda econômica nas criações de ruminantes devido à redução no ganho de peso e mortalidade de bovinos e pequenos ruminantes, especialmente em regiões com clima tropical e subtropical. A identificação precisa das diferentes espécies, bem como o conhecimento sobre a epidemiologia das gastroenterites parasitárias, são fundamentais para a elaboração de estratégias sustentáveis de profilaxia das parasitoses. Essa revisão tem por objetivo central, abordar os principais métodos parasitológicos utilizados na identificação morfológica das espécies, os quais se caracterizam pela facilidade e baixo custo. Na maioria dos estudos realizados no Brasil, a distinção entre as espécies Haemonchus contortus e Haemonchus placei não tem sido considerada. Vários relatos de H. contortus, particularmente em bovinos, podem se tratar na verdade da infecção dos animais por H. placei. A identificação correta das espécies é, portanto, fundamental. Além das medidas dos espículos dos exemplares machos, outros detalhes morfológicos, tais como a sínlofe, devem ser avaliados com o objetivo de auxiliar na diferenciação das espécies. Mensurações das larvas infectantes, obtidas em coproculturas, podem também indicar a espécie de Haemonchus presente. Esse procedimento pode ser útil especialmente em estudos que não envolvem a necropsia de animais, como é o caso de testes destinados a avaliar a resistência anti-helmíntica em rebanhos.

Moxidectin-resistant nematodes in cattle in Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Haemonchus contortus: A multiple-resistant Brazilian isolate and the costs for its characterization and maintenance for research use

The aim of this work was to determine the resistance level of Haemonchus contortus isolated from the Santa Inês flock of the Embrapa (Brazilian government's Agricultural Research Company), Southeast Livestock Unit (CPPSE), as well as to determine costs of characterizing and maintaining this isolate in host donors. Forty-two male Santa Inês lambs were experimentally infected with 4000 H. contortus infective larvae of the field isolate of CPPSE, called Embrapa2010, and divided into six treatment groups, which received triclorfon, albendazol plus cobalt sulfate, ivermectin, moxidectin, closantel and levamisole phosphate, as well as a negative control group (water). Egg per gram (EPG) counts were performed at 0, 3, 7, 10 and 14. days post treatment when the animals were slaughtered for parasite count. The data were analyzed using the RESO statistical program, considering anthelmintic resistance under 95% of efficacy. EPG and worm count presented a linear and significant relation with 94% determination coefficient. The susceptibility results obtained by RESO through both criteria (EPG and worm count) were equal, except for closantel, showing that the isolate Embrapa2010 is resistant to benzimidazoles, macrocyclic lactones and imidazothiazoles. The need of a control group did not appear to be essential since the result for susceptibility in the analyses with or without this group was the same. Suppression in egg production after treatment did not occur in the ivermectin and moxidectin groups. In the control group, the establishment percentage was just 12.5 because of the low number of third-stage larvae, resistance (innate and infection immunity) of the animals studied plus good nutrition. Drug classes presented similar efficacy between adults and immature stages. The costs for isolate characterization were calculated for 42 animals during 60. days. The total cost based on local market rates was approximately US$ 8000. The precise identification of Brazilian isolates and their establishment in host donors would be useful for laboratorial anthelmintic resistance diagnoses through in vitro tests, which has an annual cost of approximately US$ 2500 for maintenance in host donors. © 2012 Elsevier B.V.

Diagnóstico de resistência anti-helmíntica em bovinos

Pós-graduação em Medicina Veterinária - FMVZ

Resistência anti-helmíntica em equinos na Região Oeste do Estado de São Paulo

Pós-graduação em Ciência e Tecnologia Animal - FEIS

Susceptibility of helminth species from horses against different chemical compounds in Brazil

By means of parasitological necropsies, the present study aimed to evaluate, in six experiments, the degree of susceptibility or resistance of different helminth species which naturally infect horses to ivermectin 0.2mg/kg, abamectin 0.2mg/kg, moxidectin 0.4mg/kg, trichlorfon 35mg/kg, ivermectin 0.2mg/kg+praziquantel 2.5mg/kg, abamectin 0.2mg/kg+praziquantel 2.5mg/kg and ivermectin 0.2mg/kg+6.6 mg/kg pyrantel. At experimental day zero, the horses were allocated to treatment groups based on average counts of strongylid eggs per gram of feces (EPG) obtained on days -3, -2 and -1. Oxyuris sp. infections were confirmed as positive or negative. All the animals in the six experiments were naturally infected by this helminth species. Each group (control or treated) consisted of six animals. All the assessed Habronema muscae populations analyzed were susceptible to ivermectin, abamectin and moxidectin. Of the six Trichostrongylus axei populations, four were susceptible to ivermectin, abamectin, moxidectin, trichlorfon and ivermectin+praziquantel, and two were resistant to abamectin+praziquantel and ivermectin+pyrantel. Both Strongyloides westeri populations analyzed were susceptible to ivermectin, abamectin, moxidectin and abamectin+praziquantel. For O. equi, resistance was found in four different populations treated with ivermectin, abamectin, moxidectin, trichlorfon and ivermectin+praziquantel. Only combinations of abamectin+praziquantel and ivermectin+pyrantel were effective against this parasite species. All the large strongyles diagnosed in the present study (Strongyus edentatus, Strongyus vulgaris and Triodontophorus serratus) were susceptible to all the chemicals tested, with the exception of trichlorfon. Of the Cyathostominae populations, one was diagnosed as resistant to ivermectin and another to trichlorfon. The remaining populations from this nematode group were considered to be sensitive to ivermectin, abamectin, moxidectin, ivermectin+praziquantel, abamectin+praziquantel and ivermectin+pyrantel. New studies should be performed in different regions to evaluate the efficacy of trichlorfon in others field populations of helminthes.

Anthelmintic efficacy of oral trichlorfon solution against ivermectin resistant nematode strains in cattle

Infected calves from two different rural estates in Brazil were studied to assess the anthelmintic efficacy of oral trichlorfon against naturally occurring ivermectin resistant parasitic nematode strains. In experiment 1, infected animals were from a region where ivermectin resistant populations of Haemoncus placei, Cooperia punctata,Cooperia spatulata and Trichuris discolor have recently been identified. Six calves with natural gastrointestinal nematode infections were treated with 48.5 mg/kg aqueous trichlorfon administered orally and six calves acted as a non-treated control group. In experiment 11 24 naturally infected calves were selected to enter one of four treatment groups, six animals each received: 48.5 mg/kg oral trichlorfon; 200 mu g/kg subcutaneous 1% ivermectin; 630 mu g/kg subcutaneous 3.15% ivermectin; or no treatment (control group). Gastrointestinal helminths were counted and identified post-mortem at 7 days (trichlorfon and 1% ivermectin treated and untreated animals) or 14 days (3.15% ivermectin treated and untreated animals) after administration of the test agents. Experiment I identified a high level efficacy for oral trichlorfon against four helminth species that have previously been shown to be ivermectin resistant in this geographical region: percentage efficacy was 99.82% against adult H. placei, 99.18% against C. punctata, 99.33% against C. spatulata, 81.06% against T. axei, 98.46% against Oesophagostomum radiatum and 100% against T. discolor. Trichlorfon also showed activity against the ivermectin (1% and 3.15%) resistant helminth species identified in experiment 11, attaining efficacy levels of 99.17% against H. placei, 98.46% against C punctata and 100.00% against T. discolor. These findings indicate that oral trichlorfon is an effective treatment option in the management of cattle infected with ivermectin resistant helminths. (C) 2009 Elsevier B.V. All rights reserved.

Weak phenotypic reversion of ivermectin resistance in a field resistant isolate of Haemonchus contortus by verapamil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Resistance of Santa Ines, Suffolk and Ile de France sheep to naturally acquired gastrointestinal nematode infections

A study was conducted to assess the breed resistance against nematode infections in Santa Ines, Ile de France and Suffolk male lambs over a 9-month period in São Paulo state, Brazil. Lambs were born during the winter (year 2000) and were weaned at 2 months of age. The animals were then housed and treated with anthelmintics to eliminate natural infections by gastrointestinal nematodes. In late October 2000, lambs were placed in a paddock, where they stayed until August of the following year. Fecal and blood samples were taken from each animal every 2 weeks. on the same day, a pasture sample was collected to determine the number of infective larvae on the herbage. To prevent deaths, individual treatment with anthelmintics was provided to lambs with fecal egg counts (FEC) higher than 4000 eggs per gram (EPG) or with a packed cell volume (PCV) lower than 21%. In August 2001, all animals were slaughtered and the worms present in samples of the gastrointestinal contents were identified and counted. Most of the Suffolk and Ile de France sheep received three to six anthelmintic treatments over a period of 7 months, while most of the Santa Ines were not treated. Reductions in PCV and plasma protein values associated with high FEC and worm burdens were recorded, particularly, in Suffolk and Ile de France lambs. Haemonchus contortus and Oesophagostomum columbianum burdens and number of nodular lesions caused in the large intestine by O. columbianum larvae were significantly lower in Santa Ines sheep. All three breeds showed similar Trichostrongylus colubriformis worm burdens. The relative resistance of Santa Ines young male sheep was superior to that of Suffolk and Ile de France sheep. (C) 2004 Elsevier B.V. All rights reserved.

The anthelmintic effect of plant extracts on Haemonchus contortus and Strongyloides venezuelensis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

In vitro Anthelmintic effect of Melia azedarach L. and Triclinia claussenii C. against sheep gastrointestinal nematodes

The control of parasitic diseases in small ruminants is mainly done with the use of synthetic anthelmintics. However, incorrect and indiscriminate use of these products has caused the emergence of parasite resistance. Plants with anthelmintic activity are used in folk veterinary medicine, but it is necessary to investigate and scientifically validate low-cost phytotherapeutic alternatives for future use to control gastrointestinal nematodes in small ruminants by family farmers. Thus, the aim of this study was to evaluate the in vitro anthelmintic effect of plant extracts from Melia azedarach and Trichilia claussenii by the egg hatch test (EHT) and larval development test (LDT) against sheep gastrointestinal nematodes. The hexane extract of M. azedarach fruits was extracted through cold percolation and the methanol extract of T. claussenii leaves was obtained by extraction at room temperature in solvents in order of increasing polarity. The efficacy results were analyzed using the Probit program of SAS. The M. azedarach extract showed a LC50 of 572.2 mu g/mL and LC99 of 1137.8 mu g/mL in the EHT, and LC50 of 0.7 mu g/mL and LC99 of 60.81 mu g/mL in the LDT. In turn, the T. claussenii extract presented a LC50 of 263.8 mu g/mL and LC99 of 522.5 mu g/mL in the EHTand LC50 of 1.11 mu g/mL and LC99 of 26.4 mu g/mL in the LDT. Comparing the extracts of the species from the Meliaceae family, T. claussenii showed greater anti-parasite potential in vitro than M. azedarach. However, studies on the isolated compounds, toxicity and administration forms to animals are also needed to validate low-cost alternative herbal remedies for use to control gastrointestinal nematodes by family farmers. (C) 2011 Elsevier B.V. All rights reserved.