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.

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.

Anthelmintic effect of plant extracts containing condensed and hydrolyzable tannins on Caenorhabditis elegans, and their antioxidant capacity

Although tannin-rich forages are known to increase protein uptake and to reduce gastrointestinal nematode infections in grazing ruminants, most published research involves forages with condensed tannins (CT), while published literature lacks information on the anthelmintic capacity, nutritional benefits, and antioxidant capacity of alternative forages containing hydrolyzable tannins (HT). We evaluated the anthelmintic activity and the antioxidant capacity of plant extracts containing either mostly CT, mostly HT, or both CT and HT. Extracts were prepared with 70% acetone, lyophilized, redissolved to doses ranging from 1.0mg/mL to 25mg/mL, and tested against adult Caenorhabditis elegans as a test model. The extract concentrations that killed 50% (LC50) or 90% (LC90) of the nematodes in 24h were determined and compared to the veterinary anthelmintic levamisole (8mg/mL). Extracts were quantified for CT by the acid butanol assay, for HT (based on gallic acid and ellagic acid) by high-performance liquid chromatography (HPLC) and total phenolics, and for their antioxidant activity by the oxygen radical absorbance capacity (ORAC) assay. Extracts with mostly CT were Lespedeza cuneata, Salix X sepulcralis, and Robinia pseudoacacia. Extracts rich in HT were Acer rubrum, Rosa multiflora, and Quercus alba, while Rhus typhina had both HT and CT. The extracts with the lowest LC50 and LC90 concentrations, respectively, in the C. elegans assay were Q. alba (0.75 and 1.06mg/mL), R. typhina collected in 2007 (0.65 and 2.74mg/mL), A. rubrum (1.03 and 5.54mg/mL), and R. multiflora (2.14 and 8.70mg/mL). At the doses of 20 and 25mg/mL, HT-rich, or both CT- and HT-rich, extracts were significantly more lethal to adult C. elegans than extracts containing only CT. All extracts were high in antioxidant capacity, with ORAC values ranging from 1800μmoles to 4651μmoles of trolox equivalents/g, but ORAC did not correlate with anthelmintic activity. The total phenolics test had a positive and highly significant (r=0.826, p≤0.01) correlation with total hydrolyzable tannins. Plants used in this research are naturalized to the Appalachian edaphoclimatic conditions, but occur in temperate climate areas worldwide. They represent a rich, renewable, and unexplored source of tannins and antioxidants for grazing ruminants, whereas conventional CT-rich forages, such as L. cuneata, may be hard to establish and adapt to areas with temperate climate. Due to their high in vitro anthelmintic activity, antioxidant capacity, and their adaptability to non-arable lands, Q. alba, R. typhina, A. rubrum, and R. multiflora have a high potential to improve the health of grazing animals and must have their anthelmintic effects confirmed in vivo in both sheep and goats. © 2012.

Proteinograma do soro lácteo de ovelhas da raça santa inês em diferentes fases de lactação

This study aimed to evaluate the influence of lactation phases on the proteinogram of whey protein in Santa Inês ewes. Ewes were accompanied in a semi-intensive system using the same sanitary and nutritional management evaluated at 15, 30, 60 and 90 days postpartum (end of weaning and lactation). Clinical examination of the mammary gland was carried out through and bacteriological culture. The screening of the material resulted in 44 milk samples of healthy glands concurrent negative by CMT and bacteriological culture exam. For obtaining the whey protein renin solution was used. The whey was fractionated into aliquots and kept in the -80C freezer to later separation of protein fractions. For determination of total protein of whey protein was employed the biuret, observing the linearity of the test. Separation of protein fractions was performed, using polyacrylamide gel containing sodium dodecyl sulfate (SDS-PAGE). Eigth protein were observed including lactoferrin, serum albumin, IgA, IgG (heavy chain IgG (IgG CP), light chain IgG (IgG CL), ß-lactoglobulin, a-lactalbumin and proteins identified as PM 15000 and PM 29000. No significant difference was observed at different stages of lactation in the following protein: IgA (P>0.3895), lactoferrin (P>0.1611), PM 29000 (P>0.4879), α-lactalbumin (P>0.0799) and PM15000 (P>0.4494). In total protein (P<0.0022), albumin protein (P<0.0377) and IgG (P<0.0354) it was observed a significant variation in the first moments of observations, in the ß-lactoglobulin protein (P<0.0005) there was significant variation with reduction of 15 to 30 days postpartum with progressive elevation until the last stage of lactation (90 days postpartum). The SDS-PAGE technique allowed the quantification of eigth whey proteins in health ewes. The protein fractions identified reflect the profile of whey to ovine species, with influence of stages of lactation in albumin, IgG and ß-lactoglobulin.

Kinetics of epsilon antitoxin antibodies in different strategies for active immunization of lambs against enterotoxaemia

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

Técnicas de ovariectomias por videolaparoscopia em ovelhas da raça Santa Inês

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Prevalência da língua azul em ovinos da região de Araçatuba – São Paulo, Brasil

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

Avaliação da atividade de Aretmisia annua L., Melia azedarach L. e Trichilia claussenii C. sobre nematódeos gastrintestinais de ovinos

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Janaúba (Himatanthus Willd. Ex. Schult.) – Apocynaceae no controle de nematódeos gastrintestinais em ovinos

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Nematodioses de ovinos e caprinos mantidos em pastagens no oeste da Bahia

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Estudo de genômica comparativa de corynebacterium pseudotuberculosis linhagem 226 (biovar ovis)

Corynebacterium pseudotuberculosis é uma bactéria Gram-positiva, intracelular facultativa, nãoesporulante, não-capsulada e sem mobilidade, contudo possui fímbria, e pode assumir formas cocóides e filamentosas (pleomórfica), além disto, apresenta crescimento ótimo à 37°C. Este patógeno apresenta dois biovares: ovis que geralmente acomete pequenos ruminantes, e causa a doença linfadenite caseosa, e biovar equi, mais comum em equinos, bovinos, camelídeos, e bubalinos causando a Linfangite ulcerativa. A infecção por esta bactéria pode levar a condenação das carcaças e redução de lã (em ovinos e caprinos), leite e carne destes animais, e consequentemente a perdas econômicas para a indústria agropecuária mundial. Atualmente, ainda não existe uma vacina eficaz para estas doenças. A fim de obter um maior entendimento biológico entre as espécies o presente trabalho tem como objetivo principal analisar, por meio da genômica comparativa a linhagem C. pseudotuberculosis 226 biotipo ovis isolada de um caprino na Califórnia com outras linhagens do biovarar ovis e equi. Na análise de sintenia entre as linhagens foi possível identificar que a linhagem 226 apresenta alta conservação da ordem gênica entre as linhagens do biótipo ovis. Através de análises filogenômicas foi possível identificar que as linhagens I19 e 267 apresentaram maior e menor proximidade filogenética com a linhagem 226. A linhagem 1/06-A foi a que apresentou maior proximidade filogenômica entre as linhagens do biovar equi, quando comparadas a linhagem 226. Foram preditas 8 ilhas de patogenicidade, estando presente na ilha 1 os genes relacionados a virulência de C. pseudotuberculosis mais bem descritos na literatura. Não houveram regiões novas relacionadas a genes de virulência entre nenhuma das linhagens. Foram identificados 248 genes ortólogos entre as linhagens I19, 267 e 226 e 282 genes ortólogos entre as linhagens 258,1/06-A e 226. Com base nesse estudo é possível inferir que as linhagens do biovar ovis possuem um repertório gênico pouco variado e que as linhagens do biovar equi apresentam uma quantidade menor de genes compartilhados com a linhagem 226, corroborando com a diversidade gênica entre os biovares.

Alternativas para o controle de nematoides gastrintestinais de pequenos ruminantes

Sheep and goat farming requires an efficient management program, due to losses caused by parasites in susceptible animals. Many factors may collaborate to improve infection tolerance in the herd, such as: genetics, nutrition, physiological status, and age. The problem caused by resistance to antihelmintic agents has led to the spread of alternative techniques for parasite controls. The latest strategies include selective treatment with the FAMACHA method, phytotherapy, biological control with predatory fungi, and strategies that still await scientific confirmation, such as homeopathy, the dilution of resistance with the introduction of susceptible parasites, and the combination of drugs without antihelmintic effect. The main objective of these methods is to reduce the usage of antiparasitic agents, thus slowing the development of resistance and promoting the better use of effective products and newly released products. The objective of this article is to describe techniques for controlling nematodes in small ruminants, and it is aimed at technicians interested in increasing their knowledge about the mechanisms of resistance to antihelmintic agents as well as alternatives to the use of these products.

Sustainable worm control practices in South America

A high prevalence of nematodes, especially Haemonchus contortus and Trichostrongylus colubriformis, that exhibit multiple anthelmintics resistance has been reported in sheep in several South American countries. For this reason, the development of strategies that are less dependent on anthelmintic treatments is imperative for the prophylaxis of gastrointestinal nematode infections in small ruminants. Integrated grazing using cattle and sheep can be used for pasture decontamination with considerable reduction of H. contortus and T. colubnformis infective larvae after cattle grazing. Several breeds of sheep exhibit genetically related resistance against nematode infections, as is the case of crioulo, native or naturalised breeds of sheep. These breeds descend from livestock introduced by Portuguese and Spanish settlers and have been submitted to a long process of natural selection in various environmental conditions. In the South, the Crioula Lanada breed is more resistant to H. contortus than are Corriedale sheep. In tropical areas, where the minimum temperatures are usually higher than 20 degrees C, hair sheep flourish, especially the Santa Ines breed, which also display a higher level of resistance to nematode infections compared with certain breeds of European origin. However, Santa Ines sheep have inferior carcass quality compared with other commercial breeds. Recent studies showed that the crossbreeding of Santa Ines ewes with sires of breeds with high potential for growth and meat production, results in crossbred animals with high productivity and a satisfactory degree of resistance against nematode infections. Several studies have indicated that improvement in nutrition has a beneficial effect on the development of resistance in lambs that were naturally or artificially infected with nematodes. Therefore, supplementary feeding and breeding strategies to improve resistance to nematodes are feasible options in the effort to reduce dependence on anthelmintic drugs to control worm infections in sheep. (C) 2013 Elsevier B.V. All rights reserved.

Efeito da suplementação de cloreto de amônio sobre os equilíbrios eletrolítico e ácido-básico e o pH urinário de ovinos confinados

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

Two-stage rumenostomy in buffaloes

Background: Rumenostomy may be performed for therapeutic and digestibility research purposes in bovines, small ruminants and camelids. Several studies requires romenostomy in buffaloes in order to sample ruminal content for laboratorial assays. However, complications and outcome of rumenostomy was poorly studied in buffaloes. Therefore, the aim of the current study was to describe a two-stage rumenostomy technique in buffaloes, focused on intra and post-operative period.Materials, Methods & Results: Nine Murrah buffaloes were submitted to a 36-h and 12-h of food and water fastening. The animals were given acepromazine and maintained in standing position. Flank local anesthesia was carried out. A circular skin incision was carried out in the center of the left flank, followed by divulsion of the external and internal obliques and transversus abdominus muscles, and incision of the peritoneum. Subsequently, a segment of the dorsal aspect of the rumen was grasped and pulled through the flank incision. The rumen was attached to the peritoneum and skin incision margins in four points (dorsal, ventral, cranial and caudal). Additional simple interrupted sutures attaching the rumen serosa to the skin were applied subsequently. Four additional interrupted horizontal mattress sutures were applied equidistantly, taking bites only in the skin and rumen serosa. Following 12 h, the second stage was carried out. The buffaloes were prepared and restrained as performed for the first stage. A circular flap was excised from the exteriorized rumen and the silicone romenostomy cannula was placed. Clinical parameters, postoperative recovery, weight and behavioral pain scale were assessed. Positioning and anesthesia regimen were adequate for the achievement of the procedure. However, two animals fell in the restraint chute during the first surgical stage. Mild ischemia of the exteriorized rumen segment was observed on the second surgical stage, which resulted in less hemorrhage and enhanced cannula positioning. Complete cicatrization and permanent adhesion of the rumen to the skin were achieved. No ruminal leakage to the abdominal cavity occurred. No signs of pain were reported. There were few cases of laxity of the romenostomy opening leading to drop of cannula, myiasis on the margin of the stoma site and few cases of mild ruminal content leakage on the long-term assessment.Discussion: Restraint in standing position was considered adequate, although lateral recumbence constitutes another option. However, higher risk of contamination and technical difficulties in placing the cannulas are expected if lateral recumbence is considered. In other trials using acepromazine, no accidental recumbence occurred. Xylazine was also indicated for chemical restraint of buffaloes. It is known that flexible cannulas provide better anatomic adjustment and adaptation as well as being effective for sampling ruminal content, as seen in the current study. Ruminal leakage is one of the most frequent complications of romenostomy, which may affect animal's welfare. The animals in the current study presented no variations on the body score, even though on those presenting cannula loosening or ruminal content leakage. Moreover, no significant changes of the ruminal content parameters were noticed. Myiasis was also reported following ruminal surgical interventions, which were mainly attributed to extensive breeding. Loss of the cannula, subcutaneous emphysema and suture dehiscence are common complications of romenostomy. Nonetheless, none of those complications were found on the current study. Thus, romenostomy was feasible and efficient for sampling and performing assays of the ruminal content in buffaloes.