Acquisition of immunity to Eimeria maxima in newly hatched chickens given 100 oocysts

The acquisition of immunity to Eimeria maxima by chicks infected 18 hr after hatch with a single dose of 100 oocysts was investigated. In the first experiment, birds were moved each day to clean cages in order to prevent the possibility of secondary infection resulting from ingestion of oocysts passed in their feces. Immunity was measured at 4 wk of age by calculation of oocyst production following challenge with 500 oocysts or weight gain following challenge with 100,000 oocysts. Large numbers of oocysts were produced by infected birds following challenge, although numbers were significantly less than those from birds that had been reared in the absence of infection (susceptible controls). The weight gain of infected birds following challenge was significantly greater than that of susceptible controls but less than that of unchallenged controls. Thus, only partial protection had been acquired, whether parasite replication or body weight gain was used to assess the extent of immunity development. In a second experiment, acquisition of immunity at 4 wk by chicks infected 18 hr after hatch with 100 oocysts of E. maxima and reared in floor pens in contact with their droppings was investigated. Infected birds produced no oocysts following challenge, and weight gains were not significantly different from the unchallenged controls, which indicates that full immunity had developed by 4 wk. It is concluded that if oocysts of Eimeria species are used to vaccinate day-old chicks, reinfection by oocysts present in the litter is necessary for the establishment of protective immunity.     

Single and Low-level Oocyst Infections of Drug-resistant Field Strains of Eimeria tenella in Medicated Birds

Five out of ten birds infected with a single oocyst of strain Gt2 of Eimeria tenella and medicated with the recommended level of robenidine were found positive in the first experiment and four in the second in comparison with seven and six respectively in nonmedicated birds. Six birds out of ten were found positive in the two groups of similarly medicated birds infected with two or four oocysts each. Although single oocyst infections of strain Lilly 155 were unsuccessful, six out of ten birds were found positive in birds infected with ten oocysts each.All single and low-level oocyst infections were accomplished with oocysts previously treated with beta-glucuronidase and broken into sporocysts prior to infections.The overall results suggested that when a coccidium became resistant to an anticoccidial drug, only one or a few occysts were needed to start an infection if the drug was continued. The results also showed that, perhaps, successful single or low-level oocyst infections can also be used as a criterion for demonstrating drug resistance in coccidia.     

Peripartal Excretion of Eimeria Oocyst by Cows on Swedish Dairy Farms and the Age of Calves at First Excretion

Faecal samples were collected 3 times a week for 6 weeks from 22 peripartal cows and for up to 15 weeks after birth from 27 calves in 3 herds, to determine the numbers of Eimeria oocysts excreted and the age at which the calves first excreted oocysts. Only low numbers of oocysts were excreted by the cows and no oocysts were detected in 93% of the samples. However, half the cows excreted oocysts at least once. The age at which the calves first excreted oocysts ranged from 2.5 to at least 15 weeks, and there was a significant difference between the herds in their mean age at first excretion. Oocysts of Eimeria alabamensis, E. auburnensis, E. bovis and E. ellipsoidalis were found in numbers ranging from 7 to 8450 oocysts per gram faeces. About 50% of the calves excreted oocysts before they were transferred to group pens. The primary source of infection of the calves was probably their penmates or the previous occupants of the pens, and the cows probably played a subsidiary role.     

Coccidiosis in preruminating calves the effect of management and short-term treatment on the spread of infection and reinfection

In farms with large numbers of individually-housed calves, the spread of coccidia is slow. In group pens, however, all the calves became infected within 3-4 weeks of being housed together. At the beginning of group housing no oocysts were found in the faeces of any of the calves. Sulphadimidine (SDM) was administered for 3 or 12 days at different doses and different times. Administration of the drug on Days 3-5 of group housing had no effect. Given between Days 11 and 13 or 17-19, the drug lowered (for a short period) the number of animals found to be excreting oocysts. SDM given between Days 6 and 17 kept the animals oocyst-free during that period. Within 2-3 weeks after the treatment all animals were excreting oocysts.     

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.     

Experimental infection of budgerigars (Melopsittacus undulatus) with a low virulent K21 strain of Toxoplasma gondii

In total 53 budgerigars (Melopsittacus undulatus) were divided into six groups and orally infected with a suspension of oocysts of low virulent Toxoplasma gondii K21 strain in the doses of 10(2), 10(3), 10(4), 10(5) and 10(6), respectively. Blood was collected from the birds prior to the inoculation and then on days 10, 20 and 30 post infection. Latex-agglutination test (LAT) was used for the detection of antibodies in the inoculated birds. The infected birds showed no apparent signs of disease. The antibodies were found in all but two birds inoculated a dose of 10(2) oocysts. Haematological values remained unchanged after infection. T. gondii was isolated by bioassay in mice from all 37 birds fed 10(3) or more oocysts and 6 of 9 fed 10(2) oocysts. The results demonstrate that budgerigars are resistant to T. gondii infection.     

Survival of nonsporulated Toxoplasma gondii oocysts under refrigerator conditions.

Toxoplasma gondii oocysts are excreted nonsporulated in the feces of the cats into the environment. These oocysts must undergo sporulation to become infectious. Little is known about the factors that influence sporulation of T. gondii oocysts. The present study examined the survival of nonsporulated oocysts under refrigerated conditions over 11-week observation period. Microscopic examination of oocysts indicated that no visible development occurred under refrigerator conditions. The nonsporulated oocysts retained their ability to sporulate when placed at room temperature. The numbers of visually viable appearing oocysts decreased over time. Some oocysts in all samples were infectious for mice despite being refrigerated for up to an 11 weeks before undergoing sporulation. Results indicate that nonsporulated oocysts can survive in the environment for at least 3 months and retain their ability to become infectious when placed under appropriate conditions.     

The prepatent and patent periods of Eimeria alabamensis and further description of the exogenous stages

Eimeria alabamensis infections were established in calves 5 to 6 weeks of age by adminestering 10 million, 80 million, or 100 million sporulated oocysts. The prepatent period was 6 to 8 days (mean 6.6). Oocyst discharges usually lasted for 2 to 3 days although a few calves passed oocysts throughout the rest of the 3-week observation period. Calves with oocyst discharge exceeding 1 million oocysts per g of feces had a moderate diarrhea at the time of peak oocyst discharge. No other clinical signs were observed in any of the infected calves. Reinoculations with 100 million sporulated oocysts given 3 weeks after the initial inoculations of 10 million or 80 million oocysts resulted in infections characterized by greatly reduced oocyst discharges. Sporulated oocysts of E. alabamensis were 16 to 24 μ by 12 to 16 μ and were usually ovoid. The oocyst wals consisted of two layers. Sporocysts were elongate-ellipsoid, had a distinct Stieda body, and were 10 to 12 μ by 4 to 6 μ. Completely sporulated oocysts were first observed after 5 days at 25 °C, and most were sporulated after 8 days. Oocysts did not sporulate at 4 °C, 33 °C, and 37 °C.     

Experimental cryptosporidiosis and infectious bursal disease virus infection of specific-pathogen-free chickens

Specific-pathogen-free chickens orally inoculated at 4 days of age with a moderately pathogenic vaccine strain of infectious bursal disease virus (IBDV) and/or at 5 days of age with Cryptosporidium baileyi oocysts remained free of overt clinical signs throughout a 16-day period postinoculation (PI). The prepatency period for C. baileyi oocyst shedding was shorter in chickens receiving higher numbers of oocysts, but once shedding was detected, there were no obvious differences in shedding patterns among groups receiving 10(3) through 10(6) oocysts. On days 8 and 16 PI, cryptosporidia were located primarily in the bursae of Fabricius. IBDV exposure was associated with bursal follicle atrophy, whereas C. baileyi infection resulted in bursal epithelial hypertrophy and hyperplasia, mild follicle atrophy, and heterophil infiltration of the bursal mucosa. Examination of experimental groups of 30 birds each indicated that concurrent infection with both agents resulted in more severe bursal lesions, more infected birds, and greater numbers of cryptosporidia in infected tissues. At the termination of the trial, 16 days PI, Cryptosporidium infection was associated with a 6% decrease in mean body weight compared with controls.     

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.     

Elevation of muscle and plasma 3-methylhistidine as a result of turkey coccidiosis

To assess muscle breakdown during avian coccidiosis, the level of the non-metabolizable amino acid 3-methylhistidine (3MH) was determined in muscle and plasma from turkey poults that received an infection with a field isolate containing a mixture of Eimeria species. The effect of increased levels of parasitism was evaluated at 6 days postinoculation (DPI) in birds receiving 2.5 x 10(4), 1 x 10(5), or 2 x 10(5) oocysts each. The changes in 3MH levels during recovery from acute infection were assessed at 6-29 DPI in animals given 1.9 x 10(5) oocysts per bird. In some experiments, uninoculated birds given the same amount of feed as infected birds (pair fed) were used to determine the impact of feed deprivation on weight loss and 3MH levels. Infected birds had significantly elevated plasma and muscle 3MH at 6 DPI after a single dose of Eimeria oocysts. The plasma and muscle 3MH returned to control levels after 14 DPI. The 3MH levels increased with increased dose of oocysts. Plasma and muscle 3MH levels were well correlated, and an inverse curvilinear relationship between weight gain and plasma 3MH concentrations levels was observed. Plasma and muscle 3MH levels were significantly elevated in pair-fed birds, but 3MH levels in infected birds were increased by 30% over pair-fed birds. The results suggested that muscle breakdown, as assessed by plasma and muscle levels of 3MH, increased during the acute stage of Eimeria infection in turkey poults.     

Excretion of Eimeria Oocysts in Calves During their First Three Weeks After Turn-out to Pasture

The numbers of Eimeria oocysts per gram (opg) and the dry matter content of 449 faecal samples taken from 54 calves in 8 herds in south west Sweden were determined during the last 2 weeks before and the first 3 weeks after the animals were turned out to pasture. While they were housed only between 0 and 580 opg were found and in 2 of the herds the numbers of oocysts remained low after turn-out. In the other 6 herds the numbers of oocysts increased after 8 to 10 days and reached a peak of between 1080 and 80 803 opg 9 to 18 days after turnout. By 21 to 24 days after turn-out the opg-values had declined to their initial levels. Eimeria alabamensis accounted for most of the increase, but small numbers of oocysts of E. auburnensis, E. bovis, E. bukidnonensis, E. cylindrica, E. ellipsoidalis, E. pellita, E. subspherica, E. wyomingensis and E. zuernii were also observed. The interval between turn-out and the start of the increase in excretion of oocysts corresponded closely to the prepatent period of E. alabamensis and overwintered oocysts were therefore the most likely source of the infection. In 6 of the herds the dry matter content of the faeces of the calves decreased after turn-out and 56 % of the calves had clinical diarrhoea. Although it cannot be excluded that change of diet may have contributed to these symptoms, E. alabamensis infection is suggested as a potential cause of diarrhoea and loss of condition in calves in Sweden during their first weeks on pasture.     

Serologic response of red-legged partridges (Alectoris rufa) after oral inoculation with Toxoplasma gondii oocysts

Thirty 5-month-old red-legged partridges (Alectoris rufa) reared in battery were divided into five groups: 4 birds in group A, 14 birds in group B, 4 birds in group C, 4 birds in group D and 4 birds in group E, and were inoculated orally with 10, 50, 10(2), 10(3) and 10(4) oocysts of the OV-51/95 strain of Toxoplasma gondii, respectively. During the experiment, blood samples from all birds were drawn every 3-7 days and at necropsy. Serologic response was measured by the modified agglutination test (MAT) and the latex agglutination test (LAT). One bird from each group was killed at 44, 58, 65 and 72 days after inoculation (DAI). From 72 DAI to the end of the experiment, surviving partridges from group B were killed at weekly intervals. The last partridges were sacrified 100 DAI. MAT was the most sensitive and specific test for detecting T. gondii antibodies in the birds. First positive titers were detected by MAT in all sera on 7 DAI, but titers by LAT did not appear until 13 DAI. Antibody titers detected by MAT on 7 DAI were higher in the partridges with the largest inocula (10(3) or 10(4) oocysts) than those inoculated with 10, 50 or 10(2) oocysts. All surviving birds developed a serologic response to T. gondii, with maximum titers of 512-32,768 in the MAT on 13-17 DAI, and positive titers persisted at least until 100 DAI. To the contrary, LAT reveals only very low antibody titers even in partridges inoculated with the highest dose of T. gondii.     

The influence of colostrum on infection of calves around 7 months of age with Schistosoma mattheei

Thirty red-legged partridges (Alectoris rufa), 5-month-old, were orally inoculated with oocysts of the OV-51/95 strain of Toxoplasma gondii. Birds were distributed into five groups and received, respectively, 10 (group A, 4 birds), 50 (group B, 14 birds), 10(2) (group C, 4 birds), 10(3) (group D, 4 birds) and 10(4) (group E, 4 birds) oocysts. One partridge from group B and one from group E died suddenly of acute toxoplasmosis at 7 day after inoculation (DAI) with demonstrable T. gondii in several tissues. The rest of birds remained clinically normal until killed at 44, 58, 65, 72, 79 or 100 DAI. Brain, heart, liver and skeletal muscle from these partridges were bioassayed individually in mice; T. gondii was demonstrated in all these tissues, except in heart of three birds inoculated, respectively, with 10, 50 and 10(2) oocysts. Lesions were not seen in histologic sections of tissues from surviving partridges. These results suggest that red-legged partridges are resistant to clinical toxoplasmosis.     

A gel delivery system for coccidiosis vaccine: uniformity of distribution of oocysts

A patented gel delivery system being used to deliver coccidiosis vaccine to poultry hatchlings is assessed. For effective vaccination, the coccidial oocysts must be uniformly suspended before exposure to birds. The uniformity of distribution within the gel was evaluated by incorporating a culture of chicken gut flora into gel sausages, placing sections of the sausage on culture plates, determining the appearance and distribution of bacterial colonies on culture plates after incubation, and verifying by cell counts. The uniformity of distribution of similarly prepared coccidial oocysts was verified by infecting birds with 40,000 Eimeria tenella oocysts delivered via the gel. Gel-inoculated birds were compared with control birds inoculated PO with 40,000 oocysts suspended in water by using cecal lesion scores. Both the appearance and colony counts of chicken gut flora from the gel were uniform. The standard deviation in the lesion scores for the gel-inoculated group and the water-inoculated groups were 0.51 and 0.69, respectively. The results indicate that a gel delivery system can provide uniform distribution of live organisms and vaccine agents to birds.     

Efficacy of toltrazuril in broilers and development of a laboratory model for sensitivity testing of Eimeria field isolates

(1) The efficacy of toltrazuril (Baycox) against coccidiosis was established on a broiler farm in an intermittent application during five consecutive growing periods. Treated birds were fed a broiler ration without anticoccidials. The efficacy of Baycox was compared with the nicarbazin-salinomycin shuttle. It was concluded that Baycox retarded the onset of Eimeria infection for several weeks. During the fifth rearing period coccidiosis problems emerged on the farm in all birds during medication, suggesting development of resistance. (2) During a laboratory experiment the efficacy of Baycox was studied in birds after inoculation with different numbers of oocysts at 7, 10 or 15 days of age. Baycox was applied at 10 and 11 days of age. In all cases medication with Baycox protected birds from coccidiosis during a period of at least 7 days. This effect of Baycox could be due to the long-existing tissue levels of the product and its metabolites as well as its specific effect on the second generation of schizonts. (3) In another laboratory experiment coccidia obtained from field trials were tested for sensitivity to Baycox in conjunction with two strains obtained from farms were coccidiosis emerged during application. The inoculation model developed in this study was used for sensitivity testing. One of the Eimeria strains tested was resistant to the product, one strain was tolerant and the remaining two strains, including the control strain, were fully sensitive to Baycox.     

Infection by Salmonella typhimurium, S. agona, S. enteritidis or S. infantis of chicks with caecal coccidiosis

Experimental infections of Salmonella typhimurium, S. agona, S. enteritidis or S. infantis were studied in chicks infected with Eimeria tenella. In all experiments, 4-d old birds were given 5 daily oral doses of approximately 10(4) salmonella organisms per bird. One day before this inoculation one group of birds received a single dose, in the range 1 X 10(4) to 4 X 10(4) sporulated oocysts of E. tenella per os whilst the other group was not inoculated and served as the control. Chicks were examined post morten 7, 10 or 14 d after receiving coccidia. With S. typhimurium infection, the number of salmonellas in the caeca of the E. tenella-infected birds was greater than in those infected only with salmonella. With S. agona and S. enteritidis, the counts of salmonella in the caeca and the numbers of birds positive for salmonella in the caeca and liver were greater in the E. tenella-infected birds. The rate of infection of S. infantis was not increased by caecal coccidiosis.     

Subclinical coccidiosis in broilers and pullets

In the period from 1985 to 1987, 24 broiler crops (12 houses; one integration and 3 farms) and 9 pullet flocks (9 houses, 4 farms) were examined for parasites. Intestinal lesion scores and the number of parasites in the intestinal lumen (semiquantitative estimation) were recorded, and the Eimeria species determined when possible (broilers crops: 3rd and 5th week, pullet flocks 4th, 8th, 12th and 18th week). Additionally, the quantity of parasites in litter or faecal samples was examined in regular intervals. Clues for economic damage were only found in broiler crops with increased numbers of coccidial oocysts per gram litter in the 5th week of the fattening period. Eimeria tenella and E. acervulina were the dominating species in broiler chickens and also pullets, E. maxima oocysts however were only casual findings. No ectoparasites, helminths or other protozoa than Eimeria were observed. With regard to the intestinal lesions and the quantities of parasites in the intestine no significant differences were seen when comparing selected broiler chicks and birds taken at random. In pullet flocks the examination of random samples was the superior method, because only freshly dead bodies, collected in insufficient numbers, were suitable for diagnostics. In 3 broiler crops and one flock of replacement pullets kept on a wired floor no Eimeria were diagnosed.     

Intestinal digesta viscosity decreases during coccidial infection in broilers

1. The effect of intestinal digesta viscosity on bird performance in chickens with coccidiosis was compared to those without coccidiosis. 2. Six hundred chicks were divided into five groups: one control group was fed a basal maize/soyabean-based diet and the other groups were fed the basal diet supplemented with 2, 4, 6 or 8 g carboxymethyl cellulose (CMC) per kg of feed. At 14 d of age half the birds were individually inoculated with sporulated oocysts of Eimeria acervulina and Eimeria praecox. 3. Intestinal digesta viscosity increased with increasing inclusion of CMC. This effect was considerably less pronounced in inoculated than in non-inoculated birds. 4. There was a significant negative effect on live weight gain and feed conversion ratio (FCR) with increasing CMC inclusion in non-inoculated birds, but in inoculated birds there was no clear relation between CMC inclusion and performance. Neither intestinal lesion scores, nor numbers of Clostridium pefringens in the caeca, were significantly affected by CMC inclusion. 5. Across all diets inoculation impaired growth rate by 9% and FCR by 8%, but did not affect the amount of C. perfringens in the caeca.     

Evaluation of a combined immunomagnetic separation/flow cytometry technique for epidemiological investigations of Cryptosporidium in domestic and Australian native animals

A combined immunomagnetic separation (IMS) and flow cytometry (FC) technique was developed for the sensitive detection of Cryptosporidium in faecal samples. The IMS/FC technique was found to be approximately 50-fold more sensitive than formol-ether concentration, which is commonly used for Cryptosporidium epidemiological investigations. Of 31 faecal samples from captive animals 16 were found to contain Cryptosporidium oocysts when analysed using the IMS/FC compared to four when using formol-ether concentration (FEC). In a wild population of eastern grey kangaroos Macropus giganteus 66.3% of infected animals were shedding <500oocysts/gfaeces when analysed using IMS/FC. This is below the detection limit for the FEC method. The dispersal of Cryptosporidium in host populations is aggregated, with many individuals shedding low numbers of oocysts and few individuals shedding numbers of oocysts sufficiently high to be detected by FEC. This research demonstrates that the prevalence and oocyst shedding intensity of Cryptosporidium in animal populations will be significantly underestimated using standard detection methods.