Higher incidence of Eimeria spp. field isolates sensitive for diclazuril and monensin associated with the use of live coccidiosis vaccination with paracox-5 in broiler farms

Twenty European Eimeria spp. field isolates were subjected to an anticoccidial sensitivity test (AST). The anticoccidial drugs tested were diclazuril (Clinacox) and monensin (Elancoban). The assay was performed in a battery cage trial. Infected medicated birds were compared with an unmedicated control group. Coccidial lesion scores and oocyst shedding were used as parameters. The results of the AST show that resistance is common amongst coccidiosis field isolates, especially Eimeria acervulina (68% and 53% resistance for diclazuril and monensin, respectively). Resistance is less frequent amongst Eimeria maxima (38% and 50% resistance for diclazuril and monensin, respectively) and Eimeria tenella isolates (23% and 38% resistance for diclazuril and monensin, respectively). A highly significant influence of the coccidiosis prevention program (live coccidiosis vaccination with Paracox-5 vs. anticoccidial drugs in feed) on the sensitivity patterns of Eimeria spp. field isolates for both diclazuril (P= 0.000) and monensin (P= 0.001) was found. Further, when looking at the single species and each anticoccidial drug level, significantly more sensitivity of E. acervulina for monensin (P= 0.018), E. maxima for diclazuril (P = 0.009), and E. tenella for diclazuril (P = 0.007) was found in isolates originating from vaccinated flocks. Moreover, for E. acervulina and diclazuril, E. maxima and monensin, and E. tenella and monensin a trend toward higher sensitivity of isolates for these products was found when live coccidiosis vaccination was applied. The present study shows that sensitivity for the anticoccidial drugs diclazuril and monensin is more frequent in Eimeria spp. field isolates originating from broiler farms where a coccidiosis vaccination policy is followed.     

Research on avian coccidia: an update.

Despite the availability of many anticoccidial drugs, infections caused by species of Eimeria continue to be a source of significant economic loss to the poultry industry. After two decades in which the use world wide of ionophorous antibiotics gave unparalleled control of coccidiosis, drug resistance is once again tipping the balance in favour of the parasites. The realization that even the most spectacularly successful drugs might, after all, have a finite life if not used conservatively, has focused attention on ways in which the life span of drugs can be prolonged. Many drugs with different (if unknown) modes of action are available, and a variety of shuttle and rotation programmes can be considered. In view of the limitations of chemotherapy, particularly for the rearing of replacement flocks, there is considerable interest in the development of vaccines. Prospects for the introduction of live vaccines based on attenuated parasites are now very good, but the availability in the future of genetically engineered vaccines is more uncertain as little is known about the parasite molecules that stimulate protective immunity and, even if isolated, how they can be administered to the host so that it responds in the immunologically correct manner. Current research on Eimeria spp. in the chicken is broadly representative of that being done on other coccidia. Many lines of investigation are not connected with the development of new drugs or vaccination per se (and therefore have no obvious practical applications), but they are providing new insights into the biological complexity of the organisms and the ways in which they interact with their hosts. It remains possible, however, that a more detailed understanding and analysis of the molecules that are essential in the maintenance of the parasitic life style can be exploited in the future to provide alternative targets for chemical or immunological attack. The research topics considered in this review are arbitrarily grouped as studies on: (1) the basic biology of parasites, including aspects of the life cycle, and structure and function of the apical organelles; (2) the molecular biology of the parasites, including analyses of the number and structure of chromosomes, characterization of DNA sequences, and an account of the viral RNA that has been found in some species of Eimeria; and (3) control of coccidiosis, encompassing first immunity and the development of vaccines, and secondly, chemotherapy.     

Comparison of Eimeria species distribution and salinomycin resistance in commercial broiler operations utilizing different coccidiosis control strategies

The purpose of the present study was to evaluate the species composition and salinomycin sensitivity of Eimeria oocysts isolated from commercial broiler farms that differed by means of coccidiosis control (anticoccidial drugs [ACD] vs. live oocyst vaccines [VAC]). A comparison of Eimeria species composition and salinomycin sensitivity was also made before and after a producer switched from salinomycin to live oocyst vaccines. In general, no significant difference was observed in the concentration of Eimeria spp. oocysts in litter from VAC-utilizing farms compared to litter from ACD-utilizing farms. Application of PCR-based methods to detect coccidia found that Eimeria species distribution in litter from VAC operations more closely resembled the species composition in the live oocyst vaccines. Drug sensitivity testing found that Eimeria oocysts from VAC operations displayed greater salinomycin sensitivity as measured by weight gain and feed conversion efficiency compared to oocysts from ACD farms. These findings provide additional evidence for the usefulness of live oocyst vaccines to restore ionophore sensitivity in poultry operations that contain an ionophore-resistant population of Eimeria spp. oocysts.     

鸡球虫病疫苗的研究进展

鸡球虫病是由艾美耳球虫感染鸡引起的一种危害极其严重的寄生原虫病。目前,控制球虫病主要还是依赖于药物,但由于球虫耐药株的频繁出现和人们对药物残留等问题的关注,使人们更加注重探讨从免疫预防的角度去控制球虫病。本文综述了鸡球虫病活疫苗(强毒疫苗和弱毒疫苗)的研究和应用现状及重组疫苗(DNA疫苗和重组蛋白疫苗)的研究进展,同时介绍了新型疫苗佐剂的研究情况。     

Recent advances in immunomodulation and vaccination strategies against coccidiosis

Coccidiosis is a ubiquitous intestinal protozoan infection of poultry seriously impairing the growth and feed utilization of infected animals. Conventional disease control strategies rely heavily on chemoprophylaxis, which is a tremendous cost to the industry. Existing vaccines consist of live virulent or attenuated Eimeria strains with limited scope of protection against an ever-evolving and widespread pathogen. The continual emergence of drug-resistant strains of Eimeria, coupled with the increasing regulations and bans on the use of anticoccidial drugs in commercial poultry production, urges the need for novel approaches and alternative control strategies. Because of the complexity of the host immunity and the parasite life cycle, a comprehensive understanding of host-parasite interactions and protective immune mechanisms becomes necessary for successful prevention and control practices. Recent progress in functional genomics technology would facilitate the identification and characterization of host genes involved in immune responses as well as parasite genes and proteins that elicit protective host responses. This study reviews recent coccidiosis research and provides information on host immunity, immunomodulation, and the latest advances in live and recombinant vaccine development against coccidiosis. Such information will help magnify our understanding of host-parasite biology and mucosal immunology, and we hope it will lead to comprehensive designs of nutritional interventions and vaccination strategies for coccidiosis.     

鸡球虫病免疫学研究进展

鸡球虫病是严重危害养禽业的重要寄生虫病之一 ,长期以来对其防治主要依靠药物 ,并发挥了一定的积极效果。近年来因耐药虫株不断出现 ,尤其是药物残留对环境的污染及其对人类健康的威胁等一系列问题日益突出。因此 ,寻找防治球虫病的新方法是广大学者关注的焦点。在众多防治本病的措施中 ,免疫学方法因其安全、有效、无毒副作用等特点 ,倍受人们的青睐。但该方法目前仍然存在许多尚未阐明和有待解决的问题。文章对近年来鸡球虫病细胞免疫和体液免疫等方面的研究进展作了较全面系统的阐述 ,为兽医临床球虫病的有效防治提供科学的理论依据     

The efficacy and economic benefits of Supercox, a live anticoccidial vaccine in a commercial trial in broiler chickens in China

The efficacy and economic benefits of Supercox, a live anticoccidial vaccine were examined and compared with an anticoccidial drug in a trial in broiler chickens under modern commercial conditions in China. In total, 40,660 chickens were used in the present study, half of which were vaccinated with the Supercox vaccine comprising a precocious line of Eimeria tenella and non-attenuated lines of Eimeria maxima and Eimeria acervulina, and the other half were medicated with Diclazuril delivered as feed additive at the dosage of 1mg/kg of feed. The vaccine was administered orally to 7-day-old chickens. No clinical diseases were diagnosed in any of the vaccinated birds. However, clinical coccidiosis occurred in a large proportion of medicated control birds and these chickens had to be treated with anticoccidial drugs (Diclazuril and Toltrazuril). Comparison of production performance between vaccinated birds and medicated control birds revealed that the vaccine Supercox performed better than anticoccidial drugs in terms of mortalities, costs and overall economic benefits (profits). These findings demonstrated that the use of the Supercox vaccine could control clinical coccidiosis in broilers and achieve production performance superior to that using anticoccidial drugs, particularly where drug resistance might result in failure to control clinical diseases.     

Control of coccidiosis in chickens by vaccination

Control of coccidiosis in chickens has relied upon managerial measurements and the prophylactic use of coccidiostatic drugs. With the emergence of Eimeria strains that are resistant to these drugs the use and number of commercially available vaccines has increased. In this review various aspects that contribute to the development of coccidiosis are discussed, and an overview of the currently marketed coccidiosis vaccines is presented. Three groups of vaccines can be distinguished based on the characteristics of the Eimeria species included in the products: vaccines based on live virulent strains, vaccines based on live attenuated strains, and vaccines based on live strains that are relatively tolerant to the use of ionophores. These latter vaccines combine the early protective effect of ionophore treatment with the late protective effect of vaccination. The impact of future developments such as recombinant-DNA vaccines and changes in consumer's attitude towards broiler production are discussed.     

鸡球虫病免疫预防研究进展

鸡球虫病是严重危害养禽业的重要寄生虫病之一,长期以来对其防治主要依靠药物,近年来因耐药虫株不断出现,尤其是药物残留对环境的污染及其对人类健康的威胁等一系列问题日益突出,药物在鸡球虫病控制中的使用越来越受到限制。许多学者认为通过免疫方法对鸡球虫病进行防治已是大势所趋,然而由于人们对鸡体抗球虫的免疫应答过程一直不很清楚,限制了免疫学方法在抗球虫方面的应用和发展。文章综述了近年来对鸡球虫免疫原性,宿主免疫应答尤其是肠道黏膜免疫在抗球虫中的作用及球虫疫苗的研究等方面的进展,相信随着分子生物学等技术的发展,免疫预防球虫病将会有更广阔的发展前景。     

兔球虫疫苗的研究现状

兔球虫病是家兔最常见、危害最严重的疾病之一,国内外多采用抗球虫药物对兔球虫病进行预防和治疗。目前为止,我国还没有研制出商品化的兔球虫疫苗,国内外仅有少部分学者开展了兔球虫病疫苗的研究工作。主要对兔球虫疫苗活虫苗、亚单位苗、基因工程苗的研究进展进行综述,以期为兔球虫疫苗的相关研究提供参考。     

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.     

The effect of ionophore anticoccidial agents on coccidia isolated in field conditions 1984-1985

Out of six isolates of coccidia of the species Eimeria tenella, obtained in the years 1984 and 1985 on farms with a stationary occurrence of coccidiosis, five were found to have reduced sensitivity or resistance to monensin. Neither were these isolates sensitive to further monovalent ionophorous anticoccidials narasin and salinomycin. The bivalent ionophorous anticoccidial lasalocid controlled five isolates completely, but for one its effectiveness was reduced. An isolate obtained on a farm with long-term absence of coccidiosis was sensitive to all anticoccidial drugs.     

鸡球虫病弱毒疫苗的免疫机理及研究进展

鸡球虫病是由艾美耳属球虫引起的一种危害性极其严重的寄生原虫病,呈世界性分布,是最主要的鸡病之一。目前主要用抗球虫药物对其进行防制,但随着耐药虫株的产生和药物残留对肉禽品质及人体健康的影响等问题的出现,对鸡球虫活疫苗的研究成为研究热点。作者对鸡球虫弱毒疫苗的免疫机理及其鸡胚传代致弱方法、理化致弱方法和早熟选育致弱法的研究进展作一概述,并对鸡球虫弱毒疫苗的应用前景进行了展望。     

抗鸡球虫药物的合理应用

鸡球虫种类很多,目前世界公认的有8 种,其中危害最大的有艾美耳属的柔嫩艾美耳球虫、毒害艾美耳球虫2种,是养鸡业中严重多发,且防制困难的疾病之一,其死亡率达40%～50%。近几年来由于各养殖户缺乏疾病防制知识,使用药物防治球虫病时并不能做到合理选用和科学应用,导致球虫的耐药性不断增加,球虫病在一定区域仍然广泛流行。作者阐述了如何合理应用抗球虫药物来减少药物产生耐药性,使其更有效地治疗鸡球虫病。     

Responses of chickens vaccinated with a live attenuated multi-valent ionophore-tolerant Eimeria vaccine

Coccidiosis, caused by Eimeria species, is a serious economic disease of chickens (Gallus gallus) and the search for vaccines to control the disease is intensifying especially with the increasing threat of drug resistance. A live attenuated multi-valent ionophore-tolerant Eimeria vaccine has been developed that contains three ionophore-resistant Eimeria species, E. tenella, E. maxima and E. acervulina. The attenuated lines were derived from virulent field strains resistant to monensin ionophore by selection for early development in chicks. The vaccine was administered by gavage and through drinking water to broiler chickens, Chinese Yellow strain, reared in wire cages. Vaccinated medicated birds performed better than vaccinated unmedicated and medicated unvaccinated groups. The final mean weights of vaccinated medicated birds were significantly higher (P<0.05), and a better vaccine protection index, using both vaccinating methods, was achieved. Results indicated that concomitant use of ionophores and vaccines could be a useful adjunct to planned immunization in the control of coccidiosis.     

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.     

Avian coccidiosis. A review of acquired intestinal immunity and vaccination strategies

The gut-associated lymphoid tissues contain B and T lymphocytes responsible for acquired immunity to avian coccidiosis. Intestinal B cells begin producing parasite-specific antibodies shortly after infection although their role in protecting against coccidiosis is debated. T-cell-mediated immunity, predominantly by intestinal intraepithelial lymphocytes and lamina propria lymphocytes, confers the main component of protective immunity to Eimeria. Many of these cells display the CD8 and gammadelta T-cell receptor surface antigens, phenotypic markers of cytotoxic T cells. Although their role in eliminating Eimeria infection remains to be completely elucidated, T cells have been implicated in parasite transport, and their activity is augmented by interferon-gamma and interleukin-2. Because of the importance of cell-mediated immunity, coccidiosis vaccines must be capable of stimulating intestinal T cells. Orally delivered, live parasite vaccines, either unattenuated or attenuated, are powerful stimulators of intestinal cell-mediated immunity, but antigenic variability between Eimeria species present in the vaccine and in the field may restrict their commercial application. The newer generations of recombinant DNA and subunit protein vaccines, particularly when used in conjunction with interferon-gamma and interleukin-2, have shown preliminary promise in controlling experimental infections but have yet to be commercially developed.     

Use of toltrazuril in pullet breeding flocks raised on floors with anticoccidial-free feed

In 28 replacement pullet flocks with flock size of 4,000 to 25,000 birds Toltrazuril (experimental preparation, 2.5% solution, approx. 7 mg/kg body weight) was administered for a period of two days in the drinking water for metaphylactic purpose and therapeutic use, respectively. All flocks were reared without anticoccidial feed additives. Exposition to Eimeria spp. was recorded by examination of faecal samples and intestinal scrapings. Clinical flock control was achieved by vets at eight to ten day intervals. Toltrazuril was characterized by short application period, rapid mode of action, significant reduction of oocyst shedding, good compatibility and acceptance thus proving superior to conventional chemotherapy of coccidiosis. The results suggest that, even without administration of anticoccidial feed additives. Toltrazuril is an effective agent for therapeutic use in acute coccidiosis. Under insufficient management and rearing conditions repeated metaphylactic application has to be considered.     

鸡球虫入侵相关分子的研究进展

鸡球虫病是由艾美耳球虫寄生于肠道所引起的一种危害极其严重的寄生虫病,给养鸡业造成巨大的经济损失.艾美耳球虫属于顶复器门原虫,在入侵宿主细胞过程中需要通过入侵虫体顶端的顶复器分泌蛋白发挥作用.目前已报道与鸡球虫入侵相关的重要蛋白,包括微线蛋白、蛋白激酶、热激蛋白以及糖酵解酶等.这些蛋白主要参与了鸡球虫入侵宿主细胞以及在宿主细胞内的生长发育、参与了虫体的细胞周期活动以及参与了糖酵解提供虫体入侵需要的能量等,进一步对这些分子进行研究,对了解鸡球虫入侵宿主细胞的相关机理及发展抗球虫病疫苗和治疗药物将有积极的意义.     

Increased level of Eimeria sensitivity to diclazuril after using a live coccidial vaccine

Anticoccidial vaccine and an anticoccidial drug rotation program were compared to determine which program was more effective in producing coccidia populations sensitive of 1 ppm diclazuril. The study used an anticoccidial drug-sensitivity battery test (AST) to determine the baseline level of diclazuril sensitivity to field isolates of Eimeria spp. from seven broiler complexes that had used diclazuril. Based on percentage reduction in weight gain and lesion scores, 25% or fewer of the isolates were effectively controlled by diclazuril. Following the baseline sampling, four of the complexes switched to a nondiclazuril in-feed anticoccidial drug program and three of the complexes switched to a vaccination program for two broiler grow-out cycles as the sole coccidiosis-control program. This study demonstrated that the vaccine used (Coccivac-B) contained anticoccidial drug-sensitive strains. Eimeria isolates were subsequently collected from the identical houses and diclazuril AST results were compared with the baseline AST results. Following the two grow-out cycles, sensitivity of the isolates to diclazuril from the four complexes that continued to use in-feed anticoccidial drugs remained essentially unchanged. The isolates from the three complexes that switched to the vaccination program demonstrated a marked increase in diclazuril sensitivity, with 60%-100% of the isolates from each complex effectively controlled by diclazuril. Vaccination with the anticoccidial drug-sensitive strains produced a measurable increase in the level of sensitivity to diclazuril.