Demonstration of antibodies to Eimeria species in lambs by an enzyme-linked immunosorbent assay.

Soluble extracts were prepared from sporulated oocysts, unsporulated oocysts and merozoites of Eimeria crandallis, E faurei and E ovinoidalis. They were assessed for antigenicity and specificity by ELISA using rabbit antisera to sporulated oocysts or merozoites. Antibody levels were examined in sera from colostrum-deprived coccidia-free lambs, conventionally reared lambs and lambs which had received experimental infections. Maternal antibody was demonstrated in colostrum and in serum taken at 24 hours from all conventionally reared animals but not colostrum-deprived animals. Antibody levels in conventional animals dropped over the first five weeks of life and rose again during the next five weeks. Antibody was not detected in coccidia-free animals. Monospecific infections of E faurei or E ovinoidalis demonstrated antibody responses to primary and secondary infections. Some specificity of response was suggested with E faurei infection. The antigen preparations showed considerable cross-reactions between species. These serum antibody responses, although appearing too late for individual diagnosis, may assist diagnosis on a flock basis.

Ultrastructural studies of the effects of amprolium and dinitolmide on Eimeria acervulina macrogametes.

Medication of chicks with 125 ppm amprolium or dinitolmide adversely affected oocyst sporulation of Eimeria acervulina (Weybridge strain). Dinitolmide delayed oocyst production and no oocyst wall formation was seen up to 168 h post infection. Both drugs caused large numbers of abnormally small wall-forming bodies to be produced in the macrogametes. In amprolium-fed chicks, abnormal oocyst wall formation was seen. It was concluded that the main drug action was against wall forming bodies of type 2.

Avian coccidiosis: the administration of encapsulated oocysts.

Oocysts of fowl coccidia, suspended in a 12% aqueous solution of alginic acid, were dropped into 5% calcium chloride solution to form beads. Following ingestion by chicks, excystation of sporozoites was not affected. Oocyst viability was best when beads were air-dried to 33% of their original weight and stored in airtight containers at 4 degrees C. Infectivity was fully maintained for 8 weeks. Trickle infection, using beads administered orally to 2- or 3-week-old chickens, or mixed daily in food of chicks from 1-day-old, established a high level of immunity. This method of oocyst administration offers some practical advantages for the conduct of immunity and chemotherapy experiments.

Morphological effects of arprinocid on developmental stages of Eimeria tenella and E. brunetti.

Medication of chicks with 70 p.p.m. arprinocid in the food, starting 2 days prior to inoculation with Eimeria tenella, resulted in decreased oocyst production and oocyst sporulation. The main morphological alteration seen during the development of the second-generation schizont was a reduction in the number of second-generation merozoites due to incomplete merogony, resulting in large masses of vacuolated residual cytoplasm. In places around the developing macrogamete the integrity of the parasitophorous vacuole was lost by either the host cell limiting membrane making contact with the parasite pellicle, or the disruption of one or both of these surfaces apparently brought about by the intervention of the host cell mitochondria. Thirty p.p.m. arprinocid reduced sporulation of the oocysts of E. brunetti and caused enlargement of the perinuclear space in the late merozoites and swelling of the endoplasmic reticulum around wall-forming bodies II.

Problems in the identification of species of Eimeria.

The paper is concerned with the principles upon which coccidia of the genus Eimeria may be characterized. Reference strains for comparative purposes usually are not available and the limitations of morphological data for speciation are discussed. The value of other parameters are considered such as host and site specificity, pathogenicity, immunological specificity, pre-patent period, sporulation time, enzyme variation, and DNA buoyant density. The weight afforded to each of these parameters for specific identification may vary according to the parasite and host studied. Determinations of physiological and behavioral characteristics that are now becoming available should be included in species definitions wherever possible.

Observations on the epidemiology of coccidial infections in sheep under varying conditions of intensive husbandry including chemoprophylaxis with monensin.

Coccidiosis occurs sporadically in lambs at about 6 weeks of age when oocyst output is very high in healthy as well as in diseased lambs. These experiments were designed to throw light on the source of infection and to correlate oocyst output in lambs and ewes with performance in lambs. In two experiments, one indoors on deep litter the other in outdoor paddocks, oocyst output (of different coccidial species), body weight and clinical state of lambs were recorded weekly. Oocyst output in ewes was also recorded, starting 1-4 weeks before lambing. Monensin was included in the concentrates of ewes and/or lambs, up to lambing or before and after lambing. No periparturient rise was detected in the oocyst output of ewes. Monensin drastically reduced oocyst output in animals receiving it. Oocyst output in lambs appeared to be little affected by the output of ewes around the lambing period, but was reduced if the ewes' output was kept low after lambing. Lambs receiving monensin tended to produce drier faeces but their weight gain was not significantly greater than that of controls. Eimeria crandallis was the predominant species in the lambs, followed by E. ovinoidalis and E. ovina.

Freedom of coccidial oocysts from Newcastle disease virus.

Oocysts were recovered from chickens experimentally infected with the Mukteswar strain of Newcastle disease virus and Eimeria acervulina or E tenella. Oocysts sterilised by sodium hypochlorite solution were washed, ruptured and inoculated into embryonating eggs which were examined after five days by the haemagglutination test for virus. All these tests were negative. Virus, initially detected with oocysts separated from faeces, was no longer present after sporulation in 2 per cent potassium bichromate. The B1 strain of virus survived for up to 30 hours in 2 per cent sodium hydroxide and 2 per cent potassium bichromate but was quickly destroyed by sodium hypochlorite (specific gravity 1.075).

Eimeria mitis in mixed infections with E. acervulina and E. brunetti in the fowl.

Eimeria mitis and E. acervulina affect different areas of the intestine. Young chicks received E. mitis with or without E. acervulina in doses ranging from 2 to 128 X 10(4) oocysts. Mixed infections produced significantly more severe effects on growth than single infections of either species. The ratio of villus height to total mucosal thickness was reduced throughout the intestine in birds which received the mixed infection. E. mitis and E. brunetti affect the same area of the intestine. Single infections of E. brunetti in a range of doses from 1 . 25 to 20 X 10(3) oocysts produced a similar reduction in growth. When 20 X 10(3) oocysts of E. mitis were inoculated simultaneously, the effect on growth was not so severe and the oocyst output of both species was reduced. E. brunetti, superimposed 1, 2 or 3 days after an E. mitis infection, produced no greater effect on growth than E. mitis alone. Similarly, infections of E. mitis, superimposed on E. brunetti had no greater effect on growth than the latter species alone. Oocyst output of E. brunetti in superimposed infections was not reduced but peak output was delayed.

Eimeria stiedai in rabbits: the demonstration of responses to chemotherapy.

The activity of sulphaquinoxaline, robenidine, methyl benzoquate, clopidol and a mixture of methyl benzoquate and clopidol (Lerbek; Dow), was studied in rabbits infected with hepatic coccidiosis due to Eimeria stiedai. Growth inhibition, oocyst production and the activity in the serum of glutamate dehydrogenase and gamma glutamyltransferase were studied as indicators of parasite development. Only sulphaquinoxaline and Lerbek gave satisfactory control of this parasite. The latter formulation was more effective than either of its constituents used alone.

Medication against ovine coccidiosis--a review.

Data published since 1940 on the use of drugs to prevent or treat ovine coccidiosis are presented in the form of tables. These highlight the differences in conditions of husbandry and mode of administration. Such differences partly account for the variability of response to medication. Drugs that have proved useful under certain conditions include amprolium, monensin and lasalocid for prevention, and sulphonamides for treatment.

Eimeria acervulina and E. mivati: oocysts, life-cycle and ability to develop in the chicken embryo.

Three strains of Eimeria acervulina and 3 strains of E. mivati were compared. E. acervulina was characterized by the ovoidal shape of its oocysts and their rapid sporulation, small first-generation schizonts containing few merozoites, and its inability to complete development in the chorio-allantois of chicken embryos. E. mivati had more rounded oocysts with a longer sporulation time, larger first-generation schizonts containing twice as many merozoites and it developed readily in the chorio-allantois. Pre-patent periods were variable but generally shorter with E. acervulina. A strain of E. mivati attenuated by repeated embryo passage showed a reduce pre-patent period.

Ultrastructural changes in the macrogamete and early oocyst of Eimeria maxima resulting from drug treatment.

The ultrastructure of the macrogamete and developing oocyst of Eimeria maxima (Weybridge strain) was examined in the intestinal cells of chicks fed 3 different anticoccidial drugs. Amprolium at 125 p.p.m., arprinocid at 35 p.p.m. and dinitolmide at 250 p.p.m. caused considerable morphological abnormality and incomplete development of the wall-forming bodies of Type 2 (WFB II), which did not appear able to participate in oocyst wall formation. The wall-forming bodies of Type 1 (WFB I) were able in each case to participate in oocyst wall formation although amprolium and dinitolmide produced morphological abnormalities in them. In birds medicated with dinitolmide, the outer layer of the oocyst wall was formed initially at opposite poles of the macrogametes rather than as a uniform layer. Other abnormalities resulting from drug treatment are reported and some evidence that intravacuolar tubules may be formed by the parasite pellicle is presented.

Complement fixation tests for equine piroplasmosis (Babesia equi and B caballi) performed in the UK during 1976 to 1979.

The results of complement fixation (CF) test for equine piroplasmosis on sera from horses destined for international movement from Great Britain and Ireland are presented and analysed. No horses born and continuously resident in the British Isles were found carrying CF antibodies to either Babesia equi or B caballi. Positive animals were found to have association with the following countries where known tick vectors occur: Spain, Portugal, Belgium, France, Poland, USSR and Arabian Gulf countries. Data on the persistence of CF antibodies in animals subjected to repeated testing showed that some animals lost their CF titre within a few months of their first test while others retained a titre for periods up to 13 months of B caballi and 28 months for B equi. Some individuals regained titres on subsequent tests after becoming negative and the results suggest that removal from exposure to infection or, even, treatment does not necessarily eliminate a CF titre within a predictable time limit.

The chemotherapy of protozoal infections of veterinary importance.

The use of drugs in domestic animals is dominated by considerations of cost-effectiveness and profitability. They are extensively used against coccidial infections of poultry where they are an important factor in intensive husbandry. Ionophore antibiotics, which dominate this field, may have applications in ruminants. Imidocarb is of therapeutic and prophylactic value against babesial infections anf there are new prospects for control of theileriasis. Effective drugs for the control of African trypanosomiasis are limited and attention is being given to alternative uses of available compounds.

Scanning and transmission electron microscopy of Eimeria maxima microgametogenesis.

The structure and development of microgametocytes pf Eimeria maxima from chicks killed during the 6th day of infection were examined using transmission and scanning electron microscopy. Initially, nucleoli disappear and the surface of the microgametocyte invaginates. This is followed by a condensation of the chromatin which then lies at the periphery of the nuclei. Concurrent with this, the flagella are formed and they project into the parasitophorous vacuole and internal invaginations. Eventually, the nuclei become more dense and are seen to protrude from the periphery of the gametocyte. The surface view of mature microgametocytes consists of a large number of closely packed flagella.

Quantitative epidemiological studies on the prevalence of babesiosis in horses in Kuwait.

Racehorses imported into Kuwait were tested for serum antibodies to Babesia equi and B. caballi by complement fixation (CF) and indirect fluorescent antibody (IFA) tests. The prevalence of B. equi was high (77.1% by IFA) whereas that of B. caballi was much lower (11.4%). Data for B. equi showed that infection was acquired after about 6 months at risk and that in the following 12 months animals had antibody levels detectable by both CF and IFA tests by 24 months the CF reaction was no longer detectable. Estimates of incidence and inoculation rates were calculated and found to be consistent with a considerable infection pressure.

Studies with Eimeria acervulina and E. mivati: pathogenicity and cross-immunity.

Pathogenic effects caused by graded doses of recent isolates of Eimeria acervulina, E. Mivati and a new isolate, thought at first to be E. mitis, were compared. Cross-immunity studies were conducted with 3 strains of E. acervulina and 2 strains of E. mivati, together with the new isolate. The results showed marked similarities between E. mivati and the new isolate and clear distinctions between these and E. acervulina. E. mivati produced scattered petechiae but did not form discrete lesions in the intestine comparable to those seen with E. acervulina. Endogenous stages of E. mivati were most numerous in the lower small intestine and proximal caeca. The ratio of villus height to total mucosal thickness in the lower intestine was reduced. Similar changes caused by E. acervulina were seen in the anterior intestine only.