Survey of coccidial oocysts and parasite eggs in feces of free-ranging Grus japonensis

Fecal survey of Eimeria oocysts and parasite eggs was conducted for 219 fecal samples of free-ranging Grus japonensis in Kushiro district in Hokkaido in April 2003. Positive rate and mean oocysts (or eggs) per gram in positive samples were 26% (57/219) and 8.8 (0.2-136) in oocysts of Eimeria reichenowi, 18.3% (40/219) and 320 (100-1,000) in trematode eggs, 0.1% (2/219) and 0.2 (0.2-0.3) in eggs of Nematoda A, and 4.1% (9/219) and 0.8 (0.2-3.6) in eggs of Nematoda B, respectively.     

秦皇岛地区鸡球虫抗药性研究

应用艾维茵肉仔鸡，以最适抗球虫活性百分率（POAA）、病变记分减少率（RLS）和相对卵囊产量（ROP）为判定指标，检测了采自河北省秦皇岛地区的6种艾美耳球虫对8种常用抗球虫药物的敏感性。结果表明：6种混合艾美耳球虫对马杜霉素和盐霉素具有轻度抗药性；对拉沙洛菌素和氨丙淋具有中度抗药性；对球痢灵、克球多和常山酮具有重度抗药性；对地克珠利无抗药性。     

Incidence of Toxoplasma gondii oocysts in cat feces

Within two years and a half, the faeces of 620 cats coming from Brno and the area around the city were subjected to parasitological examination with special regard to the occurrence of the oocysts of Toxoplasma gondii. Sucrose solution at the specific weight of 1,150 was used as flotation medium. Oocysts of Toxoplasma gondii were eliminated by eight cats (1.29%) at the age from 16 days to 1.5 years. Six of the eight cats were younger than seven months. The Toxoplasma gondii oocysts were eliminated by the cats for 1-16 days while exhibiting signs of short-term scours and swelling of lymph nodes. In all cases the oocysts of Toxoplasma gondii were produced in the summer and autumn seasons (June-December). During the patent period, other coccidia (Isospora felis and Isospora rivolta) were also present in the cats.     

Natural infection with Eimeria leuckarti: prevalence of oocysts in feces of horse foals on several farms in Kentucky during 1986

During 1986, fecal specimens were collected 1 or more times from each of 164 horse foals (158 Thoroughbred and 6 mixed light horse type), ranging in age from 0 to 252 days, on 13 farms in central Kentucky. To detect natural infection with Eimeria leuckarti, feces were examined for oocysts. Oocysts were found in 67 (41%) of the foals on 11 (85%) of the farms. The earliest age at which oocysts were first detected was 15 days (1 foal); the latest age was 123 days (1 foal). The mean age for the first appearance of oocysts in the feces of the 67 foals positive for E leuckarti was approximately 70 days. Age of the oldest foal that passed oocysts was 185 days. The longest oocyst shedding period was about 4 months, although oocyst-positive feces usually were found only sporadically during this period.     

Column chromatographic characterization of cytoplasmic proteins in Eimeria maxima oocysts from chickens

Cytoplasmic proteins from unsporulated and sporulated Eimeria maxima oocysts were analyzed by gel-filtration column chromatography. Unsporulated oocysts were characterized as having 3 major cytoplasmic proteins and sporulated oocysts as having 5 major cytoplasmic proteins. Molecular weights ranged from 5 x 10(3) to 1.4 x 10(6). Larger molecular weight proteins were detected in sporulated and unsporulated oocysts, but were associated more with sporocysts of sporulated oocysts.     

Prevalence of oocysts of Isospora suis and Eimeria spp from sows on farms with and without a history of neonatal coccidiosis

Oocysts of Isospora suis were not found in any of 77 fecal samples from sows on farms with a history of neonatal coccidiosis. Oocysts of Isospora suis were found in 1 of 172 fecal samples from farms without a history of neonatal coccidiosis. Oocysts of Eimeria spp were found in 81.8% of the sows from farms with a history of neonatal coccidiosis and in 94.8% of the sows from farms without a history of neonatal coccidiosis. Oocysts of Isospora spp from birds were encountered as a pseudoparasite in several fecal samples.     

Immunization with Eimeria ninakohlyakimovae-live attenuated oocysts protect goat kids from clinical coccidiosis

Caprine coccidiosis, affecting mainly young goat kids around the weaning period, is worldwide the most important disease in the goat industry. Control of caprine coccidiosis is increasingly hampered by resistances developed against coccidiostatic drugs leading to an enhanced need for anticoccidial vaccines. In the current study we conducted an oral immunization trial with live attenuated sporulated Eimeria ninakohlyakimovae oocysts. Sporulated E. ninakohlyakimovae oocysts were attenuated by X-irradiation technique. The experimental design included a total of 18 goat kids divided into the following groups: (i) animals immunized with attenuated E. ninakohlyakimovae oocysts at 5 weeks of age and challenged 3 weeks later with non-irradiated homologous oocysts (group 1); (ii) animals infected with non-attenuated E. ninakohlyakimovae oocysts at 5 weeks of age and challenged 3 weeks later with non-attenuated homologous oocysts (group 2); (iii) animals primary-infected with untreated E. ninakohlyakimovae oocysts at 8 weeks of age (control of the challenge infection, group 3); (iv) non-infected control animals (group 4). Goat kids immunized with live attenuated E. ninakohlyakimovae oocysts (group 1) excreted significantly less oocysts in the faeces (95.3% reduction) than kids infected with non-attenuated ones (group 2). Furthermore, immunization with live but attenuated oocysts resulted in ameliorated clinical coccidiosis compared to goat kids infected with untreated oocysts (group 2) and resulted in equally reduced signs of coccidiosis after challenge infection compared to acquired immunity driven by non-attenuated oocysts. Overall, the present study demonstrates for the first time that live attenuated E. ninakohlyakimovae oocysts orally administered showed almost no pathogenicity but enough immunogenicity in terms of immunoprotection. Importantly, vaccinated animals still shed low amounts of oocysts, guaranteeing environmental contamination and consecutive booster infections to sustain ongoing immunity.     

Distribution of oocysts, sporocysts and sporozoites of Eimeria tenella and Eimeria maxima in the digestive tract of chicken.

The distribution of oocysts, sporocysts and sporozoites of Eimeria tenella and Eimeria maxima in the digestive tract of chicken and in excreta was investigated. At 1 h after the oral inoculation of E. tenella oocysts, the number of sporocysts in the cecum was 3.4 x 10(6) and decreased gradually thereafter, and the number of sporozoites in the cecum increased and remained at a high level until 12 h after the inoculation. Small numbers of sporocysts and sporozoites of E. tenella were found in other intestinal sites. A great number of E. maxima sporozoites was found, especially in the jejunum, 2 h after the inoculation. The findings that the largest populations of sporozoites of E. tenella and E. maxima were found in the cecum and the jejunum, respectively, indicate that the site specificity of sporozoite invasion for each species is determined before the invasion takes place.     

Characterization of proteins in sporulated and unsporulated Eimeria maxima oocysts

Proteins in sporulated and unsporulated oocysts of Eimeria maxima were characterized, using monoclonal antibodies (MAB), ELISA, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and protein (western) immunoblotting techniques. Three MAB (EM1, EM2, and EM4) were produced against proteins of sporulated oocysts. The ELISA results indicated that EM1 was reactive with sporulated oocyst proteins, EM2 was reactive with sporulated and unsporulated oocyst proteins, and EM4 was reactive with unsporulated oocysts and proteins. Separation of proteins in E maxima sporulated and unsporulated oocysts by SDS-PAGE indicated that sporulated oocysts had proteins of approximately 200 kilodaltons (kD) and distinct protein bands at 21.5 and 45 kD. Using SDS-PAGE, unsporulated oocysts had less-distinct high molecular weight protein bands (greater than 200 kD), compared with sporulated oocysts, and a distinct protein band at 31 kD. Use of all 3 MAB yielded negative results in western blot analysis of fractions obtained by SDS-PAGE.     

In vitro destruction of Eimeria oocysts by essential oils

This study aims to assess the ability of essential oils (EOs) to destroy Eimeria oocyst in vitro using microscopic counting and 273 nm absorbing material release. A screening for the ability of ten EOs to destroy Eimeria oocyst was carried out in liquid medium. Out of these ten, artemisia, tea tree, thyme and clove EOs were identified as being the most effective. The treatment of Eimeria oocyst with these EOs leads to their lysis as shown by the release of substances absorbing at 273 nm. These results were obtained after approximately three hours contact. Four EOs were proven to destroy Eimeria oocysts in a few hours at low concentration. This destructive effect is a consequence of their lysis. This work is a preliminary contribution aiming to develop a new generation of natural efficient agents for destroying Eimeria oocyst to fight coccidiosis in broiler chicken.     

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.     

Excretion of Eimeria alabamensis oocysts in grazing calves and young stock

The aims of the present study were to investigate the excretion of Eimeria alabamensis oocysts by young cattle during their first grazing season and during the first 16 days of their second grazing period. In trial 1, nine first-season grazing heifers were studied and found to have become infected with E. alabamensis shortly after turnout. The next grazing period they were turned out on to a permanent pasture together with two first-season grazing calves. Faecal samples were collected before turnout and then daily from day 3 to day 16. The second-season grazing heifers excreted insignificant numbers of E. alabamensis oocysts, whereas one of the two first-season grazing calves excreted up to 703,000 oocysts/g of faeces (OPG), indicating that the pasture was contaminated. In trial 2, faecal samples were collected from 12 calves before their first turnout in May, daily from day 2 to day 20 after turnout and then once a week until the end of September. The calves grazed pastures used in previous years by first-season grazing calves. Nine of the calves developed clinical E. alabamensis coccidiosis 4-7 days after turnout and excreted more than 950,000 OPG on days 9-10. By day 17 the oocyst excretion had decreased below 900 OPG and remained low throughout the rest of the grazing season. The results of the two studies indicate that reinfections with E. alabamensis are of little clinical importance in calves grazing contaminated pastures, and that young stock infected with E. alabamensis during their first grazing season may be used to cleanse contaminated pastures without risk of developing clinical coccidiosis.     

Differentiation of porcine Eimeria spp. by morphologic algorithms

Oocysts of Eimeria spp. were isolated from feces of naturally infected sows by conventional flotation. Saturated sodium chloride solution was superior to zinc chloride, zinc chloride/sodium chloride or sugar solution to isolate oocysts. Seven species, namely Eimeria scabra, E. polita, E. perminuta, E. debliecki, E. suis, E. porci and E. spinosa, were identified. The dimensions of oocysts (n = 4088) and sporocysts (n = 3594) were measured with an image analysis system; colour and shape of oocysts were estimated and transformed to numerical values. Of the 4088 oocysts approximately 99% were allocated to the correct species by algorithms calculated on the basis of these values. Rough-walled oocysts (E. scabra, E. polita, E. perminuta, E. spinosa) could be distinguished without previous sporulation in most cases (>97%). Smooth-walled oocysts require sporulation for further classification and were accurately allocated to species in at least 93% of cases.     

The purification of sporocysts and sporozoites from Eimeria tenella oocysts using Percoll density gradients

A protocol is presented for the purification of sporozoites from sporulated oocysts of Eimeria tenella. Two Percoll density gradients are the basis of the purification. The first gradient is used after glass-bead grinding to purify undamaged sporocysts; 87% of the sporocysts loaded onto the gradient were recovered in the pellet. The second gradient is used after excystation to purify sporozoites; 98% of the sporozoites loaded onto the gradient were recovered in the pellet. The sporozoites are 99% pure with a final recovery of about three sporozoites per oocyst.