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.     

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.     

Blackhead disease (Histomonas meleagridis) aggravated in broiler chickens by concurrent infection with cecal coccidiosis (Eimeria tenella).

The effect of concurrent cecal coccidiosis infections on severity of Histomonas meleagridis (blackhead disease) in chickens was investigated in a series of experiments. Cecal lesions from H. meleagridis were severe in all inoculated control groups and did not appear to be affected by the introduction of Eimeria tenella infection. However, the severity of liver lesions and number of birds positive for liver lesions of H. meleagridis increased significantly with the presence of E. tenella. The increase was similar when 10(3) or 10(4) oocysts of E. tenella were given and was the same when oocysts were given at the same time as H. meleagridis or 4 days prior. The liver lesions increased directly as doses of H. meleagridis increased from 7.5 x 10(3) cells to 30, 100, or 300 x 10(3) when E. tenella was given along with H. melelagridis but not when H. meleagridis was given alone. Administration of a live coccidiosis vaccine containing very low levels of E. tenella also gave a significant boost to liver lesions but at a much lower level than that observed with larger doses of E. tenella. The positive relationship between infections of cecal coccidiosis and H. meleagridis in chickens suggests that such dual exposure may contribute to increased clinical outbreaks of blackhead disease in chickens under field conditions.     

Comparison of disease susceptibility and subclass-specific antibody response in SC and FP chickens experimentally inoculated with Eimeria tenella, E. acervulina, or E. maxima

Susceptibility to disease and the subclass-specific antibody response to Eimeria tenella, E. acervulina, and E. maxima were compared in two inbred strains of chickens, FP (B15B21) and SC (B2B2). FP strain was more susceptible to coccidiosis than SC chickens based on oocyst production, lesion score, and clinical signs. FP chickens infected with E. tenella had more severe cecal lesions and a significantly lower hematocrit level than SC chickens. FP chickens infected with E. acervulina excreted five times as many oocysts at 6 days postinfection as SC and showed a 71% reduction in plasma carotenoid level compared with controls (56% reduction in SC chickens). Body-weight change did not correlate with other signs of disease. Both SC and FP chickens produced high levels of serum IgM and IgG and biliary IgA. Although SC chickens had a slightly higher antibody response than FP chickens at 7 days postinoculation, both strains maintained high levels of IgM, IgG, and IgA for a prolonged period post primary inoculation. Although SC and FP chickens show different disease susceptibility to coccidiosis, they demonstrate similar antibody response.     

Cecal coccidiosis in poultry as affected by prior exposure to aflatoxin B1.

Young New Hampshire and broiler chickens were given feed containing 2 concentrations of aflatoxin B1 (0.2 or 2.0 ppm). The larger concentration caused severe toxicosis and death in the New Hampshire chicks, but did not cause gross signs in the broiler chicks. However, temporary stunting was seen on both New Hampshire and broiler chicks during periods of aflatoxin feeding, along with persisting decreased weight gains in the broiler chicks; the latter apparently recovered during the next 21 days of feeding the starter ration. New Hampshire and broiler chicks which were given feed containing aflatoxin B1 at concentrations of 0.2 and 2.0 ppm for 28 days followed by a 21-day "recovery period" and which were not given a coccidiostat were more susceptible to severe cecal coccidiosis and had more persisting hepatic and cecal lesions than did chicks not given aflatoxin. The coccidiostat was protective against both cecal damage and losses in checks challenge exposed at 49 days of age to 100,000 infective Eimeria tenella oocysts.     

An enzyme-linked immunosorbent assay for coccidiosis in chickens: correlation of antibody levels with prior exposure to coccidia in the laboratory and in the field

An enzyme-linked immunosorbent assay was developed for detecting antibody to coccidia to facilitate the survey of laboratory and field infections. Serum antibody levels in chickens were measured against soluble Eimeria tenella oocyst antigen. Sera from breeders aged 10, 23, 37, and 43 weeks were positive with uniformly high antibody titers. Broiler chick sera showed high maternal antibody titer at hatch, decreasing to an almost negligible response at 3 weeks of age. Two-week-old broiler chicks had variable responses to a single infection of E. tenella: titers were elevated at 8 to 10 days postinfection and generally increased through day 24. Weekly reinfection of 2-week-old broiler chickens produced an antibody titer in proportion to the number of oocysts per dose and stimulated protection against challenge with 2 x 10(5) E. tenella. Inbred birds raised in a pathogen-free environment for 6 weeks had no detectable antibody titers.     

Possibilities of bacteremia and toxemia in death of chickens infected with Eimeria tenella

The possibility that bacteremia and toxemia were the causes of death in cases of cecal coccidiosis was investigated. Germ-free and ordinary chickens with microflora were inoculated with sporulated oocysts of Eimeria tenella. At 5 days postinoculation, cecal lesions in ordinary chickens were more severe than those in germ-free ones. Cardiac blood, spleen, and liver were examined in ordinary chickens for bacteremia and endotoxemia, and small numbers of bacteria were recovered from both infected and uninfected birds. Endotoxin levels in plasma of E. tenella-infected birds were low and not different from the levels of uninfected controls. To examine unknown toxic factors, a large volume of serum from infected chickens was injected intravenously into uninfected birds. No significant clinical signs were observed. It is concluded that the intestinal bacteria increase the severity of coccidial lesions without bacteremia and toxemia.     

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.     

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.     

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.     

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.     

柔嫩艾美耳球虫早熟株免疫剂量研究

为了确定柔嫩艾美耳球虫（Eimenatenella）早熟株合适的免疫剂量,本文设立7个早熟株免疫攻虫组、1个不免疫攻虫组和1个不免疫不攻虫组,免疫组的免疫剂量为孢子化卵囊100、200、400、600、800、1000和2000个／羽,经嗉囊感染,7日龄首次免疫,14日龄以同等剂量进行第二次免疫,21日龄以8×10^4个／羽的同源母株进行攻虫,28日龄结束试验,以存活率、增重、肠道病变记分、血便数量、卵囊减少率为观测指标。对免疫保护效果较好的3个免疫剂量进行重复试验,同时设置商品化球虫疫苗对照组,免疫方法、试验周期、试验指标同第一批试验。结果显示：攻虫后,不免疫攻虫组出现5％死亡,而各免疫组来出现死亡;各免疫组卵囊减少率在61．57％～69．52％;200～2000免疫组的增重与不免疫不攻虫组差异不具备显著统计学意义（P〉0．05）;600～2000免疫组的肠道病变记分和血便数量均明显少于不免疫攻虫组（P〈0．05）。用600、800和1000进行重复试验,三个免疫组攻虫期间均来出现死亡,而不免疫组和疫苗对照组均出现5％死亡;三个免疫组的增重均明显高于不免疫攻虫组和疫苗对照组（P〈0．05）;早熟株免疫组的肠道病变记分和血便数量明显低于不免疫攻虫组（P〈0．05）,而疫苗对照组与不免疫攻虫组的相当（P〉0．05）;卵囊减少率在66.30％-78．75％,高于疫苗对照组的51．82％。结果表明,该柔嫩艾美耳球虫早熟株保持了良好的免疫原性,不同免疫剂量均能诱发鸡产生免疫保护力,其中600、800和1000个／羽的免疫效果均优于疫苗对照组,可考虑以600个／羽作为该早熟株在疫苗制备中的推荐免疫剂量。     

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.     

Influence of inoculation dose, inoculation schedule, chicken age, and host genetics on disease susceptibility and development of resistance to Eimeria tenella infection

Various parasite- and host-related factors influencing disease susceptibility and development of protective immunity against Eimeria tenella infection were investigated in two inbred strains of chickens. Chickens that received a primary inoculation of 10(3), 10(4), or 10(5) oocysts showed a significant reduction in packed cell volume and produced significantly more oocysts than chickens inoculated with fewer oocysts. Younger chickens were as susceptible as older chickens to identical parasite doses. However, upon a secondary inoculation 5 weeks following primary inoculation, FP chickens 1 to 21 days old at the time of primary inoculation developed resistance to reinfection, whereas SC chickens less than 3 weeks old at the time of primary inoculation were highly susceptible to secondary infection. Flow cytometric analysis of spleen lymphocytes showed a substantial reduction in T-cell number in 1-day-old SC but not FP chickens. Furthermore, 1-week-old SC chickens showed depressed mitogenic responses to concanavalin A compared with 1-week-old FP chickens. There was no significant difference between SC and FP chickens in speen B-cell number, regardless of age.     

免疫增强剂对鸡球虫免疫保护力的作用

选取免疫增强剂卡介苗（BCG）与黄芪多糖（APS）以比较不同用法、不同攻虫强度下其增强鸡球虫免疫保护力的效果。结果表明：BCG和APS均具有明显的免疫促进作用，能更有效地抵抗5万和25万E.Tenella卵囊的攻击，但BCG效果优于APS；BCG和APS大剂量或重复使用效果均不理想；BCG的最佳使用方法为：以0.025ml／只的剂量于球虫免疫前7d使用1次。     

Immunization of young chicks using graded dose of wild strain of Eimeria tenella

A wild strain of Eimeria tenella was isolated and utilized for immunization studies. Its optimal sporulation was attained at room temperature 24-25 degrees C after 24-48 h. Two groups of chicks were immunized by dosing a graded dose of five oocysts/chick/day for 6 days followed by 50 oocysts/chick/day for 7 days. A third group was not immunized and served as a negative control. Immunized chicks gained mass at the same rate as unimmunized ones, but when challenged with 200,000 oocysts/chick, mass gains declined in the unimmunized group. The growth rate of immunized chicks was not affected by challenge (P > 0.05). Upon challenge, unimmunized chicks produced significantly more oocysts than immunized chicks (P < 0.005). Immunized chicks withstood a challenged with 200,000 oocysts/chick without developing any clinical signs whereas the unimmunized chicks developed typical clinical signs of coccidiosis. Unimmunized chicks had significantly more severe lesions in the caecum than any other group (P > 0.005) and also produced significantly more oocysts than any other group (P > 0.005).     

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。     

Coccidiosis immunization: effects of mushroom and herb polysaccharides on immune responses of chickens infected with Eimeria tenella

An experiment was conducted to investigate the effects of polysaccharide extracts (E) of two mushrooms, Lentinus edodes (LenE) and Tremella fuciformis (TreE), and an herb, Astragalus membranaceus (AstE), on the immune responses of chickens infected with Eimeria tenella. A total of 180 broiler chickens were assigned to nine groups: three groups were fed with each of the extracts (LenE, TreE, and AstE), three groups were fed with the extracts and immunized with live oocyst vaccine (LenE+V, TreE+V, and AstE+V), a group was immunized with the vaccine only, and there were two controls (E. tenella-infected and noninfected groups). The oocyst vaccine was given at 4 days of age, and the extracts (1 g/kg of the diet) were supplemented from 8 to 14 days of age. At 18 days of age, all birds except those in the noninfected group were infected with 9 x 10(4) sporulated oocysts. The results showed that at 7 days postinfection (p.i.), birds fed the extracts without vaccination had lower body weight (BW) gain than those given the vaccine only. However, the extracts in conjunction with the vaccine significantly enhanced BW gain of the infected chickens compared with the vaccine group. Of the three extracts, LenE and TreE showed a better growth-promoting effect. The extracts largely increased oocyst excretion of droppings during the primary response postvaccination. The cecal peak oocyst output and lesion scores measured at 7 days p.i. were higher in the groups fed the extracts than in the group immunized with the vaccine only, whereas those of the groups fed with the extracts and immunized with the vaccine were not significantly different from the vaccine group. Of the three extracts, both LenE- and AstE-fed groups showed lower cecal oocyst output. Thus, as compared with the extracts, the live, attenuated vaccine showed better results with significantly increased immune response in coccidial infected birds. The polysaccharide extracts may prove useful against avian coccidiosis, and, particularly when they are used in conjunction with vaccine, they have shown preliminary promise against the experimental coccidial infection.     

Low-dose immunization of northern bobwhites (Colinus virginianus) with Eimeria lettyae provides protection against a high-dose challenge

To determine whether northern bobwhite quail (Colinus virginianus) could be immunized against Eimeria lettyae by a low-dose inoculation of oocysts, we inoculated 30 birds each with either 100 or 1000 oocysts at 2 days of age (given orally by pipette). Four weeks after immunization, the immunized birds and unimmunized controls were challenged with 1 x 10(6) E. lettyae oocysts. Eight days after challenge, birds were killed and weighed, and their intestines examined for gross lesions. Effectiveness of the immunization was measured by analyzing weight gain, intestinal lesions, severity of diarrhea, feed conversion ratio, and oocyst production. After challenge, birds immunized with 100 or 1000 oocysts gained an average of 33.3 g and 28.9 g, respectively, whereas unimmunized challenged birds gained an average of 11.5 g. Immunized quail produced approximately 99.7% fewer oocysts, had minimal gross intestinal and cecal lesions, had minimal diarrhea, and had a 50% lower feed conversion ratio compared to unimmunized challenged controls. These findings indicate that vaccination is a viable option for controlling coccidiosis in quail and that further research into vaccination is warranted.     

Gel-Bead Delivery of Eimeria oocysts protects chickens against coccidiosis

Vaccines composed of either virulent or attenuated Eimeria spp. oocysts have been developed as an alternative to medication of feed with ionophore drugs or synthetic chemicals. The purpose of this study was to evaluate the use of gel-beads containing a mixture of Eimeria acervulina, Eimeria maxima, and Eimeria tenella oocysts as a vaccine against coccidiosis. Newly hatched chicks (Gallus gallus domesticus) were either sprayed with an aqueous suspension of Eimeria oocysts or were allowed to ingest feed containing Eimeria oocysts-incorporated gel-beads. Control day-old chicks were given an equivalent number of Eimeria oocysts (10(4) total) by oral gavage. After 3 days, chicks were randomly assigned to individual cages, and feces were collected between days 5 and 8 postinfection. All samples were processed for total Eimeria oocysts. At 4 wk of age, all chickens and a control nonimmunized group received a high-dose E acervulina, E maxima, and E. tenella challenge infection. Oocyst excretion by chicks fed gel-beads or inoculated by oral gavage was 10- to 100-fold greater than that of chicks spray-vaccinated with the Eimeria oocysts mixture (log 6.3-6.6 vs. log 4.8). Subsequent protection against challenge as measured by weight gain and feed conversion efficiency was significantly greater (P < 0.05) in gel-bead and oral gavage groups compared with spray-vaccinated or nonimmunized groups. Also, gel-bead and oral gavage groups showed no significant difference (P > 0.05) in weight gain and feed conversion efficiency compared with nonchallenged controls. These findings indicate that incorporation of Eimeria spp. oocysts in gel-beads may represent an effective way to deliver live oocyst vaccines to day-old chicks for preventing subsequent outbreaks of coccidiosis in the field.