杀球灵对柔嫩艾美耳球虫作用的研究

本文报道了杀球灵(Diclazuril)对鸡柔嫩艾美耳球虫E.tenella的杀灭作用、作用峰期及停药后作用持续时间。以1ppm的推荐剂量混于饲料,分别于感染前2天、感染当天、感染后1天开始饲喂,对经口一次人工感染8×10~4/只柔嫩艾美耳球虫卵囊的鸡有显著的保护作用。试验鸡不出现血便、盲肠组织学检查及粘膜刮取物镜检均不见合子及卵囊,感染后2天给药则其抗球虫作用显著降低。故可认为杀球灵作用峰期是子孢子和第一代裂殖体阶段。停药后其抗球虫作用很快减弱或消失。     

The extraintestinal stages of Eimeria tenella and E. maxima in the chicken

Donor chickens given feed medicated with one or two levels of decoquinate or given non-medicated feed were infected with oocysts of Eimeria tenella or E. maxima per os. Twelve hours after inoculation with oocysts liver, mid-intestine or ceca homogenates were fed to previously uninfected recipient chickens. The results showed that continuous medication with decoquinate was effective in preventing the transfer of sporozoites from the intestine to the liver. Oocysts were detected in the feces of all recipients of tissue from non-medicated donors, showing that some sporozoites of E. maxima and E. tenella are normally transferred to liver. Young broiler chickens were immunized by oral inoculation of E. maxima oocysts. The immune status of similar chickens inoculated with sporozoites of the same species directly into the liver or spleen were assessed. During the experimental period half of the chicks were provided with non-medicated food and the remainder were given feed supplemented with decoquinate; decoquinate was effective in arresting the development of the sporozoites. Two weeks after initial infection the birds were challenged with oocysts of E. maxima per os. Injection of sporozoites into the spleen did not protect against challenge. Birds inoculated with sporozoites into the liver were unable to develop a significant level of immunity. When the drug pressure was removed from these birds, parasitism of the intestine occurred and immunity developed.     

Protective effects of sugar cane extracts (SCE) on Eimeria tenella infection in chickens

The effects of oral administration of sugar cane extracts (SCE) on Eimeria tenella oocysts infection in chickens were studied with 2 different experiments. In Experiment 1, 3-week-old inbred chickens (MHC; H.B15) were inoculated into the crop with SCE (500 mg/kg/day) for 1 day or 3 consecutive days, and then challenged with E. tenella sporulated oocysts (2 x 10(4) cells/chicken). In Experiment 2, 1-week-old chickens were orally administered SCE at the same dose for 3 consecutive days, and then initially infected with E. tenella sporulated oocysts (2 x 10(3) cells/chicken). At 2 and 3 weeks of age, these chickens were immunized intravenously with the mixed antigens of sheep red blood cells (SRBC) and Brucella abortus (BA). At 4 weeks of age, chickens were challenged with E. tenella sporulated oocysts (1 x 10(5)/chicken). Challenged chickens with E. tenella oocysts showed markedly decreased body weight gain/day, severe hemorrhage and great number of shedding oocysts in feces and high lesion scores. Oral administration of SCE and initial infection with oocysts (2 x 10 (3)/chicken) resulted in a remarkable improvement in body weight gain/day, hemorrhage, the number of shedding oocysts and lesion score, compare to other infected groups. In addition, SCE-inoculated chickens with the initial infection showed a significant increase in antibody responses against SRBC and BA and also improvement in decreased relative proportions of Bu-1a(+) and CD4( )cells in cecal tonsil lymphocytes of E. tenella-challenged chickens. Cecal tissues of chickens administered SCE and initially infected with E. tenella oocysts showed lower numbers of schizonts, gametocytes and oocysts than those of infected control chickens. These results suggest that SCE have immunostimulating and protective effects against E. tenella infection in chickens.     

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。     

Assessment of dietary supplementation with probiotics on performance, intestinal morphology and microflora of chickens infected with Eimeria tenella

We evaluated the effects of dietary supplementation with different preparations of probiotics on the performance of broiler chickens experimentally infected with 2 × 10(4) sporulated oocysts of Eimeria tenella at 14 days of age. Three hundred, day-old, Cobb-500 chicks, as hatched, were separated into 10 equal groups with three replicates. Two of the groups, one challenged with E. tenella oocysts and the other not, were given a basal diet and served as controls without medication. The other challenged groups were given the anticoccidial lasalocid (60 mg/kg) or Enterococcus faecium (5 × 10(8) or 5 × 10(9)cfu/kg feed), Bifidobacterium animalis (5 × 10(8)cfu/kg feed), Lactobacillus reuteri (5 × 10(8)cfu/kg feed), Bacillus subtilis (5 × 10(8)cfu/kg feed), or a multi-species probiotic mix at 5 × 10(8) or 5 × 10(9)cfu/kg feed, respectively. The trial lasted 6 weeks. Individual body weight, feed intake per pen and feed conversion ratio values were recorded weekly, along with the extent of bloody diarrhea, excreta oocyst numbers and bird mortality. Caecal lesions were assessed and intestinal samples were taken for histopathological and bacteriological evaluation from ileum and caecum. Overall growth performance of chickens fed the multi-species probiotic mix at both levels was higher (P<0.05) compared to the infected control. Overall oocyst shedding was lowest (P<0.05) in the lasalocid supplemented group. Villous height was higher (P<0.05) in Bacillus supplemented groups compared to infected controls. The Lactobacillus supplemented group had the highest (P<0.05) numbers of both Lactobacillus and Bifidobacterium in ileum and caecum. In conclusion, dietary probiotics are promising for further investigation on improving intestinal health and growth performance of broiler chickens experimentally challenged with E. tenella.     

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.     

鸡胚接种柔嫩艾美耳球虫和堆形艾美耳球虫活卵囊的免疫保护试验

给18日龄鸡胚接种一定剂量的柔嫩艾美耳球虫(Eim eria tenella)和/或堆形艾美耳球虫(E.acervulina)孢子化卵囊,出雏后在无球虫环境中笼养,1~10日龄每天收集各组粪便样本,计数克粪便卵囊数(OPG),并于14日龄时以大剂量同源孢子化卵囊攻虫,以相对增重率(RWG)、饲料转化率(FCR)、相对卵囊产量(ROP)评价免疫保护效果。结果显示,以E.tenella或E.acervulina卵囊免疫18日龄鸡胚,其卵囊排出的潜隐期及达到峰值的时间与1日龄雏鸡接种组相一致,有相似的排卵囊曲线,提示其诱导免疫的建立是在出雏后开始建立的。攻虫后各免疫组的RWG由攻虫对照组的31.9%~51.7%提高到了76.5%~83.6%,RCR由攻虫对照组的4.11~4.89改善为2.72~2.96,ROP降至4.7%~23.5%。结果表明以一定剂量E.tenella和E.acervulina卵囊单独或混合经羊膜腔免疫18日龄鸡胚都可以建立起针对出雏后14日龄同源攻虫的良好免疫保护力。比较混合免疫E.tenella和E.acervulina卵囊组与单一接种E.tenella或E.acervulina卵囊组的免疫效果发现,混合免疫组的各项指标均稍优于后者。     

Studies on the stage of action of lasalocid against Eimeria tenella and Eimeria acervulina in the chicken

Broiler chickens in battery pens were either fed a diet containing 100 ppm lasalocid or no drug for 24 h prior to inoculation with sporulated oocysts of Eimeria tenella or Eimeria acervulina. Different groups of birds remained on medicated feed for 24, 48, 72, 96, 120 or 144 h after inoculation. Conversely, other groups started on an unmedicated diet, were given medicated feed at different times after oocyst inoculation. Starting lasalocid medication 24 h (E. tenella) or 48 h (E. acervulina) after inoculation reduced the lesions and improved the weight gain. There was no significant difference in performance of birds after withdrawal of the drug at 48 h (E. tenella) or 72 h (E. acervulina) and thereafter. Starting lasalocid medication at 96 or 120 h did not suppress but rather reduced oocyst production.     

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.     

Response to Eimeria tenella of chickens selected for high or low antibody response and differing in haplotypes at the major histocompatibility complex.

Sublines of chickens selected for high antibody (HA) or low antibody (LA) response that differed at the major histocompatibility complex (MHC) were tested for response to Eimeria tenella. In Expt. 1, the first exposure to E. tenella was natural (in floor pens), and chicks were challenged orally 21 days later with 0, 928, or 1855 oocysts. In Expt. 2, chicks were reared in wire-floored batteries, vaccinated orally with 928 oocysts, and challenged orally 12 days later with 15,844 oocysts. Corticosterone (20 mg/kg) was mixed with feed from 24 hr before vaccination to 120 hr after vaccination in Expt. 2. In Expt. 1, LA chicks had more-severe cecal lesions but gained relatively more body weight after challenge than did HA chicks. In Expt. 2, cecal lesions were least severe in HA chicks that had been fed corticosterone, most severe in LA chicks fed corticosterone, and intermediate in chicks that were not fed corticosterone. No differences in response to E. tenella occurred as a result of haplotypes at the MHC.     

Avian eimeria: invasion in foreign host birds and generation of partial immunity against coccidiosis

Four species of avian Eimeria invaded the intestine of foreign host birds in the same areas in which they invaded the natural host. Repeated inoculation (immunization) of chickens with the turkey coccidian, Eimeria adenoeides, partially protected the chickens against a subsequent challenge with 5.8 x 10(4) E. tenella oocysts. At 6 days post-challenge, the weight gain and feed conversion efficiency of the immunized chickens was significantly better than those of the chickens that were not immunized with E. adenoeides. Lesion scores and cellular invasion by the sporozoites were significantly lower in the immunized birds than in the unimmunized group. Electrophoresis and Western blot analysis identified changes in the serum antibody profiles of the chickens that appeared to be associated with the immunization and challenge programs. An antibody or antibodies recognizing a 60,000-molecular-weight antigen of E. tenella sporozoites disappeared when chickens immunized with E. adenoeides were challenged with E. tenella; an antibody or antibodies recognizing a 23,000-molecular-weight sporozoite antigen appeared within 6 days of challenge. Reciprocal studies, in which turkeys were immunized with E. tenella and challenged with E. adenoeides, showed little evidence of protection.     

Effects of a herbal complex against Eimeria tenella infection in chickens

A liquid and a powder made from a herbal complex consisting of Uncariae Ramulus cum Uncis, Agrimoniae Herba, Sanguisorbae Radix, Eclipta Prostrate Herba, Pulsatillae Radix, Sophorae Flavescentis Radix, Rehmanniae Radix and Glycyrrhizae Radix were studied for their anticoccidial activities in chickens. Chickens were administered with herbal liquid, powder, diclazuril or without medication during the study and challenged with oocysts of Eimeria tenella. Results indicated that the birds medicated showed less bloody faeces than those without medication. The intestinal lesion was mild in the chicks medicated with herbal liquid without significantly different lesion score when compared with uninfected chicks. The birds with medication had significantly higher body weight gains than birds without medication. Therefore, the herbal complex used in this study was effective against E. tenella infection in chickens.     

秦皇岛株和唐山株柔嫩艾美耳球虫对5种抗球虫药的抗药性检测

目的为检测柔嫩艾美耳球虫秦皇岛株和唐山株株对氯羟吡啶、氨丙啉、马杜霉素、地克珠利和尼卡巴嗪等5种抗球虫药的抗药性。方法选择280只14日龄雏鸡,随机分成14组,其中每株球虫均设5个感染用药组,1个感染不用药组,1个不感染不用药组,每组20只,以POAA、RLS、ROP、ACI作为指标。结果秦皇岛株对所选的5种抗球虫药物均产生不同程度的抗药性,而唐山株对氯羟吡啶、马杜霉素和地克珠利有不同程度的抗药性,对氨丙啉和尼卡巴嗪无抗药性。其中在这5种抗球虫药物中,秦皇岛株和唐山株柔嫩艾美耳球虫对地克珠利均呈现完全抗药性。结论目前地克珠利在秦皇岛和唐山两地区须减少或暂停使用。     

Evaluation of interspecific hybrids of the chicken, guinea fowl, and Japanese quail for innate resistance to coccidia

Experimental chicken/guinea fowl hybrids, guinea fowl, and chickens were orally inoculated with Eimeria acervulina or E. tenella, which are specific for chickens, or with E. grenieri, which is specific for guinea fowl. No intact oocysts were found in feces within 24 hr of inoculation, suggesting that excystation occurred in the normal and abnormal hosts. No oocysts were found in the feces of hybrids during a 9-day postinoculation period. The guinea fowl passed oocysts of guinea fowl coccidia (E. grenieri) but not those of chicken coccidia, and the chickens passed oocysts of chicken coccidia (E. acervulina and E. tenella) but not those of guinea fowl coccidia. Some asexual development (schizogony) occurred in hybrids inoculated with E. tenella, but sexual development (gametogony) did not. In contrast, quail/chicken hybrids became infected with oocysts of chicken coccidia (E. acervulina, E. tenella, and E. maxima) and quail coccidia (E. bateri) and passed a few oocysts during the normal patent period; control chickens and quails became heavily infected with oocysts of chicken and quail coccidia, respectively.     

Influence of reticuloendotheliosis on the severity of Eimeria tenella infection in broiler chickens

Broiler chickens free of maternal immunity to reticuloendotheliosis virus (REV) were used in the experiment. Two groups of 25 chickens were inoculated with REV at one day of age. One of these groups and another group of 25 chickens were inoculated with Eimeria tenella sporulated oocysts at 7 days of age. Chickens inoculated with E. tenella showed bloody diarrhoea from 12 to 14 days of age. Six out of 25 chickens died (P less than 0.05) at 13 and 14 days of age in the dual infected group. At 14 days of age, when chickens were killed, the lesion score in the combined infection group, was statistically different from that in the chickens inoculated with E. tenella alone. Also the weights of the bursa of Fabricius and thymus were lower in the two REV infected groups than in the controls. Although REV infection alone adversely affected the weight gain and feed conversion, with combined infection this effect was much greater. Following REV inoculation most of the chickens showed feathering defects and all the examined chickens were viraemic at 21 days of age. At the same age, all but one chicken failed to show precipitating antigenaemia and about one-half of these chickens showed a very low serum neutralisation titre. None of these chickens showed precipitating antibodies.     

柔嫩艾美耳球虫地克珠利抗药株的实验室诱导

采用药物浓度递增法,以0．07mg／L地克珠利为起始诱导浓度连续传代,以最适抗球虫指数（POAA）、病变记分减少率（RLS）和相对卵囊产量（ROP）3项指标综合判定抗药性。经12次传代,该诱导虫株对1．5mg／L地克珠利具有完全抵抗力。试验结果表明,用实验室方法诱导柔嫩艾美耳球虫抗药株完全可行。     

Isolation and selection of ionophore-tolerant Eimeria precocious lines: E. tenella, E. maxima and E. acervulina

Eimeria parasites were isolated from Nanhai Guangdong province (southern China) and studied in chickens in wire cages to evaluate their drug resistance against commonly used ionophores: monensin (100 mg/kg of feed), lasolacid (90 mg/kg), salinomycin (60 mg/kg), maduramicin (5 mg/kg) and semduramicin (25 mg/kg). Chinese Yellow Broiler Chickens were infected with 40,000 crude sporulated Eimeria oocysts at 15 days of age and prophylactic medication commenced a day prior to infection. Drug resistance was assessed for each ionophore drug by calculating the anticoccidial index (ACI) and percentage optimum anticoccidial activity (POAA) based on relative weight gain, rate of oocyst production and lesion values. Results revealed that Nanhai Eimeria oocysts comprising of E. tenella, E. maxima and E. acervulina, were resistant to monensin, sensitive to both salinomycin and lasolacid and partially sensitive to maduramicin and semduramicin. By selection for early development of oocysts during passage through chickens, the prepatent time of E. tenella, E. maxima and E. acervulina were reduced by 49, 36 and 22 h, respectively. The precocious lines are less pathogenic than the parent strains from which they were selected and conferred a satisfactory protection for chickens against coccidiosis. These ionophore-tolerant precocious lines could have wider applications in the development of anticoccidial vaccines for sustainable control of coccidiosis.     

Development of protective immunity against Eimeria tenella and E. acervulina in White Leghorn chickens inoculated repeatedly with high doses of turkey coccidia

Repeated inoculation (immunization) of 2-week-old white leghorn chickens with 10(6) oocysts of the turkey coccidia Eimeria adenoeides or E. meleagrimitis partially protected chickens against moderate challenge with E. tenella or E. acervulina oocysts, but not with E. necatrix oocysts. After challenge, mean weight gains of the immunized chickens and the unchallenged controls did not differ significantly, but weight gains of unimmunized chickens were significantly lower. The mean feed-conversion ratio of the immunized challenged chickens was 3.14, as compared with 4.42 for unimmunized challenged control chickens. In general, immunization did not markedly reduce intestinal lesions. Repeated inoculation of chickens with the turkey coccidium E. gallopavonis failed to produce statistically significant protection against challenge with E. tenella, E. acervulina, or E. necatrix, as determined by weight gain, feed-conversion efficiency, and lesion scores. Antibody profiles of individual chickens did not correlate with protection.     

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。     

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.