Garlic as an Alternative Anthelmintic in Sheep

Abstract

An increase in the anthelmintic resistance of parasites in small ruminants and a push towards non-chemical farming has lead researchers to search for alternative forms of anthelmintics. The efficacy of treating sheep by using natural garlic extract with respect to changes in weight, fecal egg counts (FEC), and packed cell volume (PCV) was investigated. Sheep were treated every 4 weeks for 8 weeks with saline (control), Ivermectin, or natural garlic extract and samples were collected weekly to determine FEC and PCV. Sheep were weighed at the time of sample collection to determine weight change. There were no significant differences between the Ivermectin and natural garlic extract treated sheep with respect to weight changes, FEC or PCV. The Ivermectin and natural garlic extract treated sheep had significantly greater (P < 0.05) weight changes and PCV compared to the control. The control sheep had a significantly higher (P < 0.05) FEC compared to the Ivermectin and natural garlic extract treated sheep. Administering natural garlic extract as anthelmintic is a variable alternative to Ivermectin. INTRODUCTION Small ruminant gastrointestinal nematodes potentially reduce production and cause profit loss throughout the sheep industry (Perry and Randolph, 1999). Traditionally, producers have used commercially available anthelmintics (benzimidazoles, imidazothiazoles-tetrahydropyrimidines, and avermectins-milbemycins) to control parasites. However, an increase in the anthelmintic resistance of parasites is currently a serious threat and research is being done to find alternative forms of anthelmintics (Larson, 2006). The societal push towards non-chemical (eco-friendly, green, organic) farming has also hastened the search for viable alternatives to chemical anthelmintics (Waller and Thamsborg, 2004). Haemonchus Contortus, often referred to as the "barber pole" worm, punctures the lining of the abomasum, causing blood plasma and protein loss in sheep. Since it is usually the most prevalent nematode parasite in sheep and causes the most destruction, a majority of the research in alternative forms of anthelmintics has been targeting H. Contortus. Copper oxide wire particles administered to sheep have been shown to reduce the number of H. Contortus strongyle eggs shed in lambs (Burke et al., 2004) as well as pregnant ewes (Burke et al., 2005). Developing vaccinations against parasites is becoming a reality with the use of recombinant protein-based vaccines (Knox, 2000), however the economical availability of such vaccines to producers is Corresponding Author: B. D. Whitaker, University of Findlay, 1000 1 North Main Street, Findlay OH, 45840, USA. E-mail: whitaker@findlay.edu Virginia Journal of Science, Vol. 61, No. 1, 2010 http://digitalcommons.odu.edu/vjs/vol61/iss1 4 VIRGINIA JOURNAL OF SCIENCE currently not a possibility. Additional research has shown that feeding forages high in tannin content reduces the fecal egg counts (FEC) and number of worms in goats (Shaik et al., 2006). Strong evidence suggests that using chlamydospores (nematode ingesting fungi) as an anthelmintic in sheep is very successful (Fontenot et al., 2004) however it has not been shown to target H. Contortus. Although not an alternative treatment, research indicates that breeding and selecting for ewes with higher genetic merit for growth as lambs, and lambs with higher genetic merit for body weight, were all more resistant to infection as adults (Vanimisetti et al., 2004). Any novel treatment needs to easily be incorporated into a producers flock management and be economically feasible (van Wyk et al., 2006). The current techniques available to test the efficacy of any treatment are serology testing, the FAMACHA chart based on the color of the eye membrane mucosa (measures relative levels of anemia) (van Wyk and Bath, 2002), and FEC (Cringoli et al., 2004). Novel treatments and management systems should be able to be evaluated using the above techniques. Therefore, the objectives of this study were to compare the use of natural garlic extract to Ivermectin as an anthelmintic for sheep with respect to, 1) change in body weight (BW), 2) FEC, and 3) packed cell volume (PCV) based on serology analysis. MATERIALS AND METHODS Experimental Site and Treatment Groups This study was carried out at the Ferrum College Agriculture Center in Ferrum, Virginia (36o92'N), between May and September 2009. The experimental site was at an altitude of 430 m and consisted of a 0.5 ha plot containing a clean fescue and orchard grass mix. Free access to unlimited water was available throughout the study. A total of 14, 90-d sheep (American black-faced) ranging from 18 to 32 kg (average = 26 kg) were randomly assigned to receive orally either saline (control), natural garlic extract (Garlic Barrier, Glendale CA, 1 teaspoon/head), or Ivermectin (Merial Ivomec, Atlanta GA, 42 mg/kg bodyweight). All sheep were treated on week 0 and week 4 of the 9 week study. Samples were collected each week for analysis. Body Weight Measurements Sheep were weighed weekly using an electronic scale (A and A Scales LLC, Prospect Park, NJ, USA) to monitor changes in BW. Fecal Egg Counts Feces were taken rectally using the gloved hand method and stored at 4< C until analysis. All FEC were performed using the McMaster method (Cringoli et al., 2004). Briefly, approximately 4.0 g fecal material was placed in a 50 mL graduated cylinder and Sodium Nitrate (1.2 – 1.25 specific gravity) was added to bring to final volume to 26 mL. The solution was homogenized, filtered through 2 layers of cheesecloth, and immediately 1.0 mL of resulting solution was placed on a McMaster slide. The slide was incubated at room temperature for 5 min to allow the eggs to float to the top of the solution and strongyle eggs were counted at 100X using a compound light microscope. Virginia Journal of Science, Vol. 61, No. 1, 2010 http://digitalcommons.odu.edu/vjs/vol61/iss1 GARLIC ANTHELMINTIC FOR SHEEP 5 Immune Response Whole blood was obtained via jugular venipuncture (BD Vacutainer, 12 mg EDTA, Franklin Lakes, NJ) at stored at 4

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大蒜对绵羊的替代驱虫药作用

小型反刍动物体内寄生虫对驱虫药耐药性的增强，以及对非化学农业的推动，促使研究人员寻找替代形式的驱虫药。研究了天然大蒜提取物对绵羊体重、粪蛋数(FEC)和堆积细胞体积(PCV)的影响。每4周给羊进行生理盐水(对照)、伊维菌素或天然大蒜提取物治疗，连续8周，每周采集样品测定FEC和PCV。在采集样品时称量绵羊体重，以确定体重变化。伊维菌素和天然大蒜提取物处理的绵羊在体重变化、FEC和PCV方面没有显著差异。伊维菌素和天然大蒜提取物处理绵羊的体重变化和PCV显著高于对照组(P < 0.05)。对照绵羊的FEC显著高于伊维菌素和天然大蒜提取物处理的绵羊(P < 0.05)。施用天然大蒜提取物作为驱虫药是伊维菌素的可变替代品。小型反刍胃肠道线虫可能会降低整个绵羊产业的产量并造成利润损失(Perry和Randolph, 1999)。传统上，生产者使用市售的驱虫药(苯并咪唑、咪唑噻唑-四氢嘧啶和阿维菌素-米贝霉素)来控制寄生虫。然而，寄生虫对驱虫药耐药性的增加目前是一个严重威胁，正在进行研究以寻找替代形式的驱虫药(Larson, 2006)。社会对非化学(生态友好、绿色、有机)农业的推动也加速了寻找化学驱虫药的可行替代品(Waller和Thamsborg, 2004)。弯曲血蜱，通常被称为“理发杆”蠕虫，会刺穿皱胃的内壁，导致绵羊的血浆和蛋白质流失。由于它通常是绵羊中最普遍的线虫寄生虫，并且造成最大的破坏，因此大多数替代形式的驱虫剂的研究一直针对H. Contortus。给羊注射氧化铜丝颗粒已被证明可以减少小羊羔(Burke et al.， 2004)和怀孕母羊(Burke et al.， 2005)的圆形圆孔圆孔卵的数量。利用重组蛋白为基础的疫苗开发针对寄生虫的疫苗正在成为现实(Knox, 2000)，然而这种疫苗对生产者的经济可用性是。通讯作者:B. D. Whitaker, University of Findlay, 1000 1 North Main Street, Findlay OH, 45840, USA。E-mail: whitaker@findlay.edu Virginia Journal of Science, Vol. 61, No. 1, 2010 http://digitalcommons.odu.edu/vjs/vol61/iss1 4 Virginia Journal of Science目前不可能。进一步的研究表明，饲喂高单宁含量的饲料可减少山羊的粪蛋数(FEC)和蠕虫数量(Shaik et al.， 2006)。强有力的证据表明，在绵羊中使用衣孢子(线虫摄取真菌)作为驱虫剂是非常成功的(Fontenot et al.， 2004)，但尚未显示其针对H. Contortus。虽然不是一种替代治疗方法，但研究表明，培育和选择具有较高遗传优势的母羊作为羔羊生长，以及具有较高体重遗传优势的羔羊，成年后对感染的抵抗力都更强(Vanimisetti et al.， 2004)。任何新的处理方法都需要容易地纳入生产者群体管理，并在经济上可行(van Wyk et al.， 2006)。目前可用于测试任何治疗效果的技术是血清学测试、基于眼膜粘膜颜色的FAMACHA图表(测量贫血的相对水平)(van Wyk和Bath, 2002)和FEC (Cringoli等人，2004)。新的治疗和管理系统应该能够使用上述技术进行评估。因此，本研究的目的是通过血清学分析，比较天然大蒜提取物与伊维菌素作为绵羊驱虫剂在以下方面的差异:1)体重(BW)变化;2)FEC变化;3)细胞体积(PCV)。材料与方法实验地点和实验组本研究于2009年5月至9月在弗吉尼亚州Ferrum的Ferrum学院农业中心(36o92n)进行。试验场地海拔430 m，由0.5 ha的地块组成，地块中含有干净的羊茅和果园草混合。在整个研究过程中，可以免费获得无限量的水。共有14只90天的羊(美国黑脸羊)，体重从18到32公斤(平均= 26公斤)，随机分配口服生理盐水(对照组)、天然大蒜提取物(大蒜屏障，格兰代尔CA, 1茶匙/头)或伊维菌素(梅里亚伊芙米克，亚特兰大，42毫克/公斤体重)。在为期9周的研究中，所有羊在第0周和第4周接受治疗。 每周采集样本进行分析。每周使用电子称(A and A Scales LLC, Prospect Park, NJ, USA)称羊体重以监测体重变化。用手戴手套法从直肠取下粪便，保存在摄氏4度以下，待分析。所有FEC均采用McMaster方法进行(Cringoli et al.， 2004)。简单地说，将大约4.0 g粪便放入50 mL刻度圆筒中，加入硝酸钠(1.2 - 1.25比重)使最终体积达到26 mL。将溶液均质，通过2层粗棉布过滤，并立即将1.0 mL所得溶液置于麦克马斯特载玻片上。在室温下孵育5分钟，使卵浮到溶液顶部，使用复合光学显微镜以100倍计数圆形卵。全血通过颈静脉穿刺获得(BD Vacutainer, 12 mg EDTA, Franklin Lakes, NJ)，保存在4<C保存直到分析。PCV的测定采用造血比容管。实验采用完全随机设计，数据采用SAS (SAS Institute, Cary, NC, USA)的一般线性模型(GLM)程序进行分析。模型的影响包括治疗和时间。对时间没有显著影响，从最终模型中删除。采用最小二乘均值(LSMEANS)语句和可能概率值(PDIFF)选项分析处理间的显著性。在所有分析中，P < 0.05被认为是显著的。结果用最小二乘平均值±s.e.m表示。体重测量天然大蒜提取物处理的绵羊体重变化(0.23±0.17 kg)与伊维菌素处理的绵羊体重变化(0.36±0.15 kg)无显著差异。天然大蒜提取物和伊维菌素处理的绵羊明显比对照组重(-0.26±0.17 kg) (P < 0.05)(图1)。在给药前第一周，各处理组之间的粪蛋数量无显著差异。在研究的其余时间里，天然大蒜提取物处理的绵羊FEC(1805.0±613.0个鸡蛋/g粪便)与伊维菌素处理的绵羊FEC(863.0±548.0个鸡蛋/g粪便)无显著差异。对照绵羊的FEC(4993.0±613.0个蛋/g粪便)显著高于天然大蒜提取物和伊维菌素处理的绵羊(P < 0.05)(图2)。免疫反应在研究的第一周和第二周，任何处理组之间的PCV均无显著差异。在接下来的研究中，天然大蒜提取物处理的绵羊的PCV(25.5±1.1%)与伊维菌素处理的绵羊的PCV(27.5±1.0%)没有显著差异。天然大蒜提取物和伊维菌素处理绵羊的PCV显著高于对照绵羊(18.9±1.1%)(P < 0.05)(图3)。本文的结果比较了在绵羊寄生虫管理中使用市售的驱虫剂(伊维菌素)和使用非化学替代品(天然大蒜提取物)的情况。随着绵羊生产者继续看到其羊群中抗驱虫寄生虫的增加，越来越需要找到替代目前做法的方法(Larson, 2006年)。我们的研究结果表明，在与商业驱虫药相同的时间指导下口服天然大蒜提取物(大蒜屏障)，与商业处理的绵羊相比，体重、FEC和PCV没有显著差异。弗吉尼亚科学杂志，第61卷，第1期，2010 http://digitalcommons.odu.edu/vjs/vol61/iss1 6弗吉尼亚科学杂志每4周给药一次驱虫剂对8周后粪卵数(FEC)(卵/克粪便)的影响。数据以平均值±SEM表示。不同上标的均值差异至少P < 0.05。*图1。每4周给药一次对8周后体重变化(kg)的影响。数据以平均值±SEM表示。不同上标的均值差异至少P < 0.05。*《弗吉尼亚科学杂志》，2010年第61卷第1期http://digitalcommons.odu.edu/vjs/vol61/iss1绵羊用大蒜驱虫剂7在整个实验过程中，我们观察到用天然大蒜提取物和用伊维菌素的绵羊的体重没有显著变化。与其他治疗组相比，未接受驱虫药治疗的羊体重减轻，很可能是由于寄生虫负荷的增加。 这对生产者是有利的，因为他们可以只使用天然大蒜提取物，而不是市售的大蒜提取物

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