Anti-protozoal efficacy of medicinal herb extracts against Toxoplasma gondii and Neospora caninum

The purpose of this study was to determine whether alcohol extracts of herbs (Sophora flavescens Aiton, Sinomenium acutum (Thunb.) Rehder and E.H. Wilson, Pulsatilla koreana (Yabe ex Nakai) Nakai ex T. Mori, Ulmus macrocarpa Hance and Torilis japonica (Houtt.) DC.) from South Korea, possess in vitro anti-protozoal activity against cultures of Toxoplasma gondii and Neospora caninum. These herbs have been used as human anti-parasitics in Asian countries for many years. Alcohol extracts of these herbs were serially diluted to final concentrations ranging from 625 to 19.5 ng/ml in media and added to wells containing either T. gondii or N. caninum tachyzoites in equine dermal (ED) cells. Parasite growth inhibition was measured using 3H-uracil incorporation as compared to untreated controls. T. japonica inhibited T. gondii proliferation by 99.3, 95.5, 73.0 and 54.0% in the range from 156 to 19.5 ng/ml, and S. flavescens inhibited T. gondii proliferation by 98.7, 83.0 and 27.2% in the range from 156 to 39 ng/ml. T. japonica inhibited N. caninum proliferation by 97.8, 97.9, 85.3 and 46.4% in the range from 156 to 19.5 ng/ml. S. flavescens inhibited N. caninum proliferation by 98.6, 97.0, 69.5 and 14.0% in the range from 156 to 19.5 ng/ml. Toxicity to host cells was noted when concentrations of T. japonica and S. flavescens exceeded 625 ng/ml. The herb extracts from S. acutum, Pulsatilla koreana, and U. macrocarpa also showed toxicity at higher levels but did not achieve the same inhibition effects at the lower concentrations against T. gondii and N. caninum as T. japonica and S. flavescens.     

白头翁不同提取物及复方抗鸡柔嫩艾美耳球虫研究

以存活率、相对增重率、盲肠病变记分、卵囊值和抗球虫指数为判定指标,考察白头翁不同提取物及复方对鸡柔嫩艾美耳球虫孢子化卵囊致病性的影响(体外试验)和抗鸡柔嫩艾美耳球虫感染效果(体内试验)。体内、体外试验结果均表明,白头翁不同提取物及复方均能提高相对增重率,降低盲肠病变计分和卵囊值。抗球虫指数(ACI)检测表明,白头翁不同提取物及复方对球虫孢子化卵囊有微弱的抑杀效果,对鸡柔嫩艾美耳球虫感染预防效果较差,不宜作为鸡柔嫩艾美耳球虫病的预防和治疗药物。     

青蒿散抗鸡球虫药效研究

为探讨青蒿散对鸡柔嫩艾美耳球虫（E.tenella）的防治效果,本试验选取90只健康黄羽肉鸡,分为6组,除空白对照组外,其余各组（感染对照组、药物对照组及青蒿散低、中、高剂量组）试验鸡均经口感染6×10⁴个柔嫩艾美耳球虫孢子化卵囊,各试验组在接种前一天开始用药至试验结束。记录各组试验鸡的眼观病变、组织病理学检查、相对增重率、盲肠病变记分、血便记分、每克粪便中的卵囊数（OPG）、卵囊值及抗球虫指数（ACI）指标,评价青蒿散不同给药浓度的抗球虫效果。结果显示,空白对照组、感染对照组、药物对照组及青蒿散低、中、高剂量组的抗球虫指数分别为200.00、72.23、157.19、82.89、125.32和137.06,与感染对照组相比,药物对照组和青蒿散各剂量组的眼观病变、组织病理学检查均有所缓解,青蒿散能减轻病鸡盲肠的肿胀和出血等症状,且病鸡精神状态较感染对照组好;与感染对照组相比,各给药组的平均增重均有所升高,盲肠病变、卵囊值及血便情况减少。综上所述,青蒿散能缓解柔嫩艾美耳球虫侵染鸡盲肠所致的病理症状和组织病变,减少粪便中卵囊的排出数量,有一定的抗球虫效果,且给药浓度越高抗球虫效果越好,具有剂量依赖性。     

Guar meal ameliorates Eimeria tenella infection in broiler chicks

Guar meal contains relatively high levels of saponins, which are known to have antiprotozoal activity and may be effective against coccidiosis. A 2x2 factorial experiment investigated the impact of guar meal (0 or 5%) corn-soy-based starter broiler diets on chicks unchallenged or challenged with Eimeria tenella. At 1 day of age, 120 unsexed RossxRoss broiler chicks were randomly distributed among four treatment groups. Chicks were challenged with 5x10(3) sporulated oocysts of E. tenella in 0.5ml at 10 days of age by oral gavage. Weekly body weight, body weight gains, feed conversion ratio and mortality rate were recorded for chicks fed from 0 to 21 days of age. Oocysts shed per gram feces were recorded from 6 to 10 days post-challenge. Results showed that challenged chicks fed 0% guar meal had significantly higher oocysts per gram shed in feces than the other groups. No significant differences among treatment groups in mortality rate were observed. Body weights of unchallenged and challenged chicks fed 0% guar meal were significantly higher than those fed 5% guar meal at 2 weeks of age. Results indicated that including 5% guar meal in the diet of chicks challenged with E. tenella decreased oocysts shed per gram feces and prevented bloody diarrhea, but without affects on body weight and feed conversion ratio at 11 days post-challenge.     

Effect of dietary supplementation with oregano essential oil on performance of broilers after experimental infection with Eimeria tenella.

A study was carried out to examine the effect of dietary supplementation of oregano essential oil on performance of broiler chickens experimentally infected with Eimeria tenella at 14 days of age. A total of 120 day-old Cobb-500 chicks separated into 4 equal groups with three replicates each, were used in this study. Two groups, one infected with 5 x 10(4) sporulated oocysts of E. tenella and the other not, were given a basal diet and served as controls. The other two groups also infected with E. tenella were administered diets supplemented with oregano essential oil at a level of 300 mg/kg, or with the anticoccidial lasalocid at 75 mg/kg. Following this infection, survival rate, bloody diarrhoea and oocysts excretion as well as lesion score were determined. Throughout the experimental period of 42 days, body weight gain and feed intake were recorded weekly, and feed conversion ratios were calculated. Two weeks after the infection with E. tenella supplementation with dietary oregano oil resulted in body weight gains and feed conversion ratios not differing from the non-infected group, but higher than those of the infected control group and lower than those of the lasalocid group. These parameters correspond with the extent of bloody diarrhoea, survival rate, lesion score and oocyst numbers and indicated that oregano essential oil exerted an anticoccidial effect against E. tenella, which was, however, lower than that exhibited by lasalocid.     

Direct effects of Moringa oleifera Lam (Moringaceae) acetone leaf extract on broiler chickens naturally infected with Eimeria species

Avian coccidiosis is one of the most important diseases of poultry and it is responsible for a large number of all broiler mortalities worldwide. The control of this disease relies mainly on the use of anticoccidial drugs. However, herbal preparations could be an alternative for the treatment against coccidiosis in chickens. The direct effects of Moringa oleifera acetone extracts on broiler chickens naturally infected with mixed Eimeria species was investigated to determine the relative efficacy of the extracts against coccidiosis in birds. The investigations were carried out in seven groups (ten chickens per group). The birds were given various doses (1.0, 2.0, 3.0, 4.0 and 5.0 g/kg body weight) of acetone extract of leaves of M. toltrazuril® (positive control) and untreated (negative control). The extract was evaluated for anticoccidial activity by means of inhibition of oocyst output in faeces, faecal score, weight gain and mortality. Haematological indices were evaluated by standard methods. The group treated with 1.0 g/ kg body weight Moringa oleifera extract produced the least inhibitory effect on oocyst shed in the faeces (96.4 %), while the groups treated with 2.0 g/kg, 3.0 g/kg, 4.0 g/kg and 5.0 g/kg body weight of the extract produced 97.4, 98.7, 99.1 and 99.8 %, respectively. Body weight gains of infected chickens treated with the extract significantly improved (p?<?0.05), and faecal scores were milder. Packed cell volume, haemoglobin concentration and red blood count of the treated birds were significantly (p?<?0.05) higher than those of the infected untreated group. Moringa oleifera leaves could find application in the treatment of avian coccidiosis in veterinary practice.     

三株柔嫩艾美球虫野外分离株的致病性分析

用每鸡4×104和8×104个孢子化卵囊的剂量感染黄羽肉雏,以临床症状、肉眼病变、平均增重、病变记分和死亡率为判定指标,对柔嫩艾美球虫EtCZ-1、EtCZ-2、EtCZ-3株的致病性进行了比较分析。结果显示:各虫株引起的临床症状和肠道肉眼病变基本相似,致病性随着感染剂量的增加而增强。各感染组的增重与对照组相比均有显著差异(P<0.50),相对增重率分别下降了74.4%～83.2%、68.7%～72.2%和70.6%～74.4%;感染后鸡出现血便的时间较一致,即第108小时开始排血便,在120h～144h达高峰;病变值和卵囊值均随着感染剂量的增加而增大;除EtCZ-1株高剂量组的存活率是90%以外,其余各组均为100%。显示3株球虫的致病性相似。     

Anticoccidial effects of <Emphasis Type="Italic">Aloe secundiflora</Emphasis> leaf extract against <Emphasis Type="Italic">Eimeria tenella</Emphasis> in broiler chicken

Anticoccidial effects of Aloe secundiflora crude leaf extract was tested in broiler chickens following oral infection with Eimeria tenella. Sixty 22-day-old birds were divided into six groups of ten birds each. Three treatment groups A, B, and C were fed with the extract (100, 250, and 500 mg/day, respectively) mixed in feed for 10 days, and three control groups: group D (drug control) administered 300 mg/l of sulfachloropyrazine sodium soluble powder in drinking water for 5 days, group E (infected/non-medicated positive control), and group F (uninfected/non-medicated negative control). Except for group F, all groups were orally inoculated with 75,000 sporulated oocysts of E. tenella. The effects of the extract on E. tenella infection were evaluated by severity of bloody diarrhea, body weight (BW) gain, oocyst output, and lesion score. No bird in the treated groups died of coccidiosis, and severity of bloody diarrhea was milder than in the positive control group. BW gains in the treated groups were significantly higher than in group E (p < 0.05). The lesion scores of the treated groups were significantly lower than that of group E. Oocyst output in groups A, B, and C were 11.23, 8.24, and 6.82 × 10⁶, respectively. As compared with the negative control group (12.84 × 10⁶), the reductions in oocyst production were 12.54, 35.83, and 46.88%, respectively. Oocyst output significantly reduced with an increase in Aloe dosage. The findings of this study suggest that Aloe secundiflora extract presents an alternative anticoccidial agent for the control of avian coccidiosis.     

Drug resistance in coccidia: a robenidine-resistant strain of eimeria tenella.

Tests for resistance to a recently introduced anticoccidial drug, robenidine, were performed on a strain of Eimeria tenella recovered from broiler chickens infected with acute cecal coccidiosis. The strain was identified previously and again confirmed by the timing of mortality of the infected chickens (five to six days postinfection), the appearance of lesions and parasites in ceca only, and the measurements (21.6 +/- 1.9 micron X 19.8 +/- 2.4 micron) of the oocysts. The tests showed that the strain of E. tenella could establish infections in chickens medicated with fourfold the recommended level of robenidine in feed but no oocysts could be recovered from the feces of infected chickens when the level of drug was increased to eightfold. The decrease in mortality, in the severity of lesions and oocyst output and the increase in the average weight gain of infected chickens followed closely the increase in the level of robenidine in feed. In contrast, no infections were found in chickens infected with a sensitive strain of E. tenella and maintained on feed mixed with the recommended level of robenidine. This and other findings discussed here show that resistance to robenidine is developing in coccidia commonly found in broiler houses.     

Prophylactic and therapeutic effects of a novel granulated formulation of Artemisia extract on broiler coccidiosis

Purpose

Coccidiosis is an important parasitic disease in poultry industry. Owing to the development of drug resistance against Eimeria and concerns about drug residues, attentions toward the alternative compounds including herbal medicines have been increased. This study aimed to examine the prophylactic and therapeutic effects of a new formulation derived from Artemisia sieberi extract on avian coccidiosis.

Methods

The extract was obtained from A. sieberi using petroleum ether, and then, it was formulated into a wet granule. Three hundred and sixty 1-day-old broilers were divided into six groups, each with three replicates (n?=?20); the first group was chosen as the noninfected control group and the remainders were challenged by oral administration of 250,000 oocysts of Eimeria tenella per chick on day 21. Group 2 (as nontreated control) received no treatment, but group 3 (as prophylactic group) received 0.5 mg/kg artemisinin 10 days before challenging with E. tenella. However, groups 4, 5, and 6 received 1 mg/kg artemisinin on the next day and at 24 and 48 h after the challenge, respectively, for 5 days. Mortality rate, bloody diarrhea score, lesion score, and oocysts per gram (OPG) of feces were used to determine the anticoccidial effects of the formulation.

Results

The granule significantly (p?<?0.05) reduced mortality, diarrhea, and lesion score in the treated groups. The OPG output was also significantly (p?<?0.0001) reduced in the groups that received formulation. However, there was no significant difference between the prophylactic and therapeutic groups.

Conclusion

The results suggested that this new formulation is a promising herbal medicine that can be used as a prophylactic or therapeutic product to control avian coccidiosis.     

球抗预防鸡柔嫩艾美耳球虫病效果的观察

采用接种孢子化柔嫩艾美耳球虫卵囊的方法,复制鸡球虫病,观察并计算球抗对柔嫩艾美耳球虫病鸡的存活率、相对增重率、血便记分、盲肠病变记分和卵囊值的影响,并求出球抗的抗球虫指数.结果发现球抗能提高球虫病鸡的存活率和相对增重率,减少血便记分和盲肠内容物卵囊数,减轻盲肠病变.表明球抗对鸡柔嫩艾美耳球虫病有一定的预防作用.     

中草药提取物对多重耐药猪胸膜肺炎放线杆菌体外抑菌试验及抑菌效果研究

为筛选对猪接触传染性胸膜肺炎放线杆菌多重耐药菌株有较好抑菌作用的中草药。挑选了穿心莲、黄芩、苦参、白头翁、芒果叶和百里香6种中草药,采用水提取和65%乙醇提取相结合的方法制备单剂中草药的提取液,然后浓缩至含原生药物1g/mL。采用二倍稀释法分别测定这6种中草药对传染性胸膜肺炎放线杆菌多重耐药菌株的最小抑菌浓度(minimal inhibition concentration, MIC)和最小杀菌浓度(minimal bacteriocidal concentration, MBC)。以筛选出体外抑菌效果较好的中草药。结果表明,胸膜肺炎放线杆菌多重耐药菌株抑菌圈最大的是百里香和芒果叶,其抑菌圈直径分别为40 mm和35 mm.,其次为白头翁、黄芩、穿心莲和苦参,其抑菌圈直径在8~16 mm。百里香、芒果叶、白头翁、黄芩、穿心莲和苦参的最小抑菌浓度分别为0.198 mg/mL、0.198 mg/mL、0.396 mg/mL、0.396 mg/mL、25 mg/mL和25 mg/mL。百里香、芒果叶、白头翁、黄芩、穿心莲和苦参的最小杀菌浓度分别为0.396 mg/mL、0.396 mg/mL、0.792 mg/mL、0.792 mg/mL、50 mg/mL和50 mg/mL。说明除穿心莲和苦参外,其余4种中草药对胸膜肺炎放线杆菌多重耐药菌株有很多好的抑菌和杀菌效果,其中百里香和芒果叶效果最佳。本结果为由传染性胸膜肺炎放线杆菌多重耐药菌引起的猪接触传染性胸膜肺炎的防控和治疗奠定了基础,为开发中草药相关无抗产品以及防治由多重耐药菌引起的猪接触传染性胸膜肺炎提供了参考。     

纳川珠利对鸡柔嫩艾美耳球虫病的防治效果研究

以存活率、相对增重率、血便计分、病变值、卵囊值和抗球虫活性指数(Anticoccidial index,ACI)为药效评判指数,考察了新三嗪类化合物纳川珠利对人工感染的鸡柔嫩艾美耳球虫病的预防和治疗效果.结果显示,纳川珠利对鸡柔嫩艾美耳球虫病有良好的防治效果,ACI值均属高效范围,有望开发为新的抗球虫药物.     

杀球灵对鸡盲肠球虫病的治疗试验及作用峰期探讨

本试验用１０日龄公雏９６只,分为８个组。氯嗪苯乙氰用量为１×１０－６,其中６个组分别在感染后０,４８,７２,９６,１２０,１５４ｈ给药治疗人工感染１×１０５个柔嫩艾美耳球虫孢子化卵囊的雏鸡。结果表明杀球灵（Ｃｌｉｎａｃｏｘ）为高效抗球虫药物,兼有预防和治疗作用。作用于盲肠球虫的有性生殖和无性生殖阶段。     

中药对柔嫩艾美尔球虫感染鸡外周血T细胞亚群的影响

为研究中药对柔嫩艾美尔球虫感染鸡免疫水平的影响,本实验采用流式细胞技术检测柔嫩艾美尔球虫感染鸡外周血CD4+T细胞亚群,以及CD8+T细胞亚群数目的动态变化。结果表明,接种球虫后,感染给中药组鸡CD4+、CD8+T细胞亚群数目及百分比(CD4+T/CD8+T)均高于感染不给药组,9d时,感染给中药组最高,分别为9.77和7.53,与感染不给药组差异显著(p0.05)。感染球虫组CD4+T/CD8+T比值均低于不感染组,感染给中药组在接种后第6d和第9d均高于感染不给药组。感染给中药组、不感染给中药组和不感染不给药组CD4+T/CD8+T平均值均在2-1之间,分别为1.11、1.33和1.25,而感染不给药组仅为0.93。     

Influence of toltrazuril treatment on parasitological parameters and health performance of piglets in the field--an Austrian experience

Porcine coccidiosis caused by Isospora suis is one of the leading causes of neonatal diarrhea in suckling piglets. Currently the only registered drug for metaphylaxis is toltrazuril. To evaluate the effect of treatment on piglets from 7 Austrian farms without and 8 Austrian farms with toltrazuril application we examined oocyst excretion (including determination of oocysts per gram of feces; OPG), diarrhea (fecal score FS 1-4 with 3 and 4 being diarrhea), and general health (health score HS 1-4 with 3 and 4 describing poor health). Both groups included farms with different levels of hygiene. Samples from 265 litters without treatment, comprising 1588 individual samples, and 1548 samples from 258 treated litters were taken twice (around the 14th and the 21st day of life, respectively), examined by autofluorescence and, if positive, by McMaster counting. In both groups animals had less diarrhea and lower health scores during the second sampling but the treated piglets were always significantly healthier and had less diarrhea. The percentage of weaned piglets was higher in treated animals although this was not significant (p=0.052). In the first round of sampling 17.8% of the individual samples from untreated piglets were positive for oocysts (with a maximum prevalence on the 12-15th day of life) while in the treated piglets only 0.4% shed oocysts p<0.001). At the second sampling only 2.1% of the untreated animals and none of treated piglets excreted I. suis (p=0.083). Positive animals shed up to 8 × 10(3)OPG. There was an increased risk for infected piglets to develop diarrhea (odds ratio, OR 4.73) and poor health (OR 5.05) in untreated piglets, and poor hygiene without disinfection was identified as a risk factor for poor health (OR 1.90), diarrhea (OR 1.42) and oocyst excretion (OR 1.73). The risk of poor health (OR 2.89) and diarrhea (OR 1.44) was also increased for piglets under poor hygienic conditions receiving toltrazuril, so both metaphylaxis of coccidiosis and good hygiene are necessary to effectively control neonatal diarrhea. The costs of treatment are considerably lower than the estimated financial production losses. Therefore, treatment is recommended for farms where clinical coccidiosis is diagnosed.     

免疫调节剂与抗球虫药联合应用的抗球虫效果

为探讨免疫调节剂对鸡球虫病的免疫反应调节作用,设计并进行了免疫调节剂与抗球虫药合用的效果的研究。试验结果以抗球虫指数的测定来判定抗球虫药与免疫调节剂合用的效果。结果表明,免疫调节剂与抗球虫药合用状态下,抗球虫指数检测显示免疫调节剂卡介苗和黄芪多糖可明显提高抗球虫指数、改善球虫感染鸡的临床症、提高增重速度等,为控制球虫病提供了新的途径。     

Anti-protozoal efficacy of high performance liquid chromatography fractions of Torilis japonica and Sophora flavescens extracts on Neospora caninum and Toxoplasma gondii

We previously reported that alcoholic extracts of Sophora flavescens and Torilis japonica from South Korea demonstrated good efficacy in reducing replication of Toxoplasma gondii and Neospora caninum. To characterize the chemical component associated with anti-protozoal activity, specific fractions were isolated by high performance liquid chromatography (HPLC) and used for in vitro testing. These fractions were evaluated in vitro against T. gondii and N. caninum. Fractions of the herb extracts were serially diluted to final concentrations of 2.850 to 0.356 ng/ml in medium and added to wells containing replicating T. gondii and N. caninum. To determine the ability of each fraction to inhibit parasite proliferation, 3H-uracil incorporation was used to determine parasite replication. In cultures infected with T. gondii, a fraction of T. japonica (TJ2) inhibited T. gondii proliferation by 99.2, 94.4, 88.6 and 27.0% in the range from 2.850 to 0.356 ng/ml. Four fractions of S. flavescens (SF1-SF4) inhibited T. gondii proliferation by 99.6-60.6, 96.9-48.1, 92.3-68.2 and 95.4-52.9% in the range from 2.850 to 0.356 ng/ml. In cultures infected with N. caninum, a fraction of T. japonica (TJ2) inhibited N. caninum proliferation by 98.3, 95.5, 79.7 and 30.6% in the range from 2.850 to 0.356 ng/ml. Four fractions of S. flavescens (SF1-SF4) inhibited N. caninum proliferation by 97.1-25.9, 94.8-35.5, 95.9-33.7 and 95.4-49.4% in the range from 2.850 to 0.356 ng/ml. These fractions of T. japonica and S. flavescens extracts are currently undergoing in vivo evaluation in experimentally infected mice.     

Coccidiostatic action of monensin fed to lambs: body weight gains and feed conversion efficacy.

Thirty crossbred ewe lambs weighing an average of 37.3 kg were allotted to 6 groups of 5 lambs each so that group weights were nearly equal. Lambs were fed dehydrated alfalfa pellets, initially at 1.14 kg/day and subsequently increased after experimental day 15 and 42. Each lamb was artificially infected with 18,000 sporulated oocysts of Eimeria ninakohlyakimovae. Groups 1, 2, 3, and 4 were given monensin in the form of medicated alfalfa pellets at dose levels of 5, 10, 20, and 30 g/metric ton, respectively. Groups 5 and 6 were infected controls (infected, nonmedicated). Lambs in groups 5 and 6 developed severe clinical coccidiosis, having diarrhea and losing weight rapidly. Group 1 lambs did not have diarrhea, but the lambs did not gain well. All other medicated lambs gained weight during the experimental period of 84 days. Feed conversion was good in medicated groups 2, 3, and 4 and was poor in control groups 5 and 6. Statistically significant differences in feed conversion and body weight gains (5 and 1% level of confidence) were observed between control and medicated groups.