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Antibodies to coccidia: detection by the enzyme-linked immunosorbent assay (ELISA).

The ELISA test was used for the detection of antibodies to coccidia in the serum and/or egg yolk of chickens infected with Eimeria acervulina, E. maxima or E. tenella and in the serum of rats infected with E. nieschulzi. Antigens prepared from different developmental stages of the parasite were tested and the cross-reaction between different species of Eimeria were examined. The variability in cross-reactivity of different species and the advantages and possible applications of the test are discussed.

Animals↗

Efficacy of antibiotics A204 sodium A28695A, and A204np against Coccidia of rabbits.

Three polycyclic, polyether, monocarboxylic acid antibiotics produced by and extracted from a strain of Streptomyces albus and designated A204 sodium, A28695A, or A204-np, were specifically tested for activity against hepatic and incidentally against intestinal coccidia of rabbits. All were effective in preventing infections and acceptable to young rabbits when prepared in pelleted feed. Weight gain, however, was not as good as inoculated, medicated rabbits as in noninoculated, nonmedicated controls. No gross pathologic changes were detected except in the livers of inoculated, nonmedicated controls which were severely infected on greatly enlarged. Livers of medicated rabbits were uninfected and normal in appearance and size.

Animals↗

Coccidia (Protozoa: Eimeridae) of the herbivorous mole-rat, Spalax leucodon Nordmann.

A survey of 96 herbivorous mole-rats, Spalax leucodon Nordmann, in Lalahan district of Ankara, Turkey, revealed the presence of 6 new species of coccidia. The species found, the shape and average length-width dimensions (in micrometers) of their oocysts, and the numbers of animals from which they were isolated were as follow: Eimeria lalahanensis sp. n., subspherical to ellipsoidal (20.3 X 12.5), from 10; Eimeria tuzdili sp. n., spherical to subspherical with radially striated oocyst wall, (20.5 X 16.5), from 3; Eimeria spalacis sp. n., ellipsoidal to cylindrical (16.3 X 9.8), from 90; Eimeria elliptica sp. n., ellipsoidal (14.3 X 10.1), from 8; Eimeria turkmenica sp. n., ellipsoidal (11.3 X 8.9), from 1; Isospora anatolicum sp. n., spherical (7.6 X 5.1), from 1 animal. The overall infection rate for 96 examined herbivorous mole-rats was 100%.

Animals↗

Eimeria chihuahuaensis sp.n. and other coccidia from Dipodomys spp. in El Paso County, Texas.

Dipodomys merriami Mearns and Dipodomys ordi Woodhouse were surveyed for coccidia in El Paso County, Texas. Infections with Eimeria chobotari, Eimeria dipodomysis and Eimeria balphae were 24.8%, 4.4%, and 11%, respectively, for D. ordi. Dipodomys merriami had an infection level of 23.8% with E. chobotari. Four animals concurrently harbored E. chobotari and E. balphae or E. dipodomysis or a new species Eimeria chihuahuaensis. Male and female host infection levels were not significantly different. The new species is described and photographs of 3 previously described Eimeria from Dipodomys are presented.

Animals↗

Coccidia (Apicomplexa: Eimeriidae) from sciurid rodents (Eutamias, Sciurus, Tamiasciurus spp.) from the western United States and northern Mexico with description of two new species.

Since May 1979, 190 rodents in the family Sciuridae, representing three genera and nine species, have been collected in the western United States and northern Mexico and examined for coccidia; 71 (37%) had coccidian oocysts in their feces. These included 2 of 12 (17%) Eutamias canipes; 7 of 12 (58%) E. dorsalis; 18 of 50 (36%) E. merriami; 33 of 96 (34%) E. obscurus; 3 of 4 (75%) E. townsendii; 3 of 9 (33%) Sciurus aberti; 1 of 1 S. griseus; 1 of 1 Tamiasciurus hudsonicus mogollonensis; and 3 of 5 (60%) T. mearnsi. The following coccidians were identified from infected rodents: Eimeria cochisensis n. sp. and Eimeria dorsalis n. sp. from E. canipes, E. cochisensis, E. dorsalis, and E. tamiasciuri from E. dorsalis, E. dorsalis and E. tamiasciuri from E. merriami; E. cochisensis, E. dorsalis, E. tamiasciuri, and E. wisconsinensis from E. obscurus; E. cochisensis and E. dorsalis from E. townsendii; E. ontarioensis and E. tamiasciuri from S. aberti; E. tamiasciuri from S. griseus; E. tamiasciuri and E. toddi from T. h. mogollonensis; and E. tamiasciuri from T. mearnsi. Sporulated oocysts of Eimeria dorsalis n. sp. were ovoid, 21.9 x 16.8 (17-24 x 14-20) micrometer with sporocysts ovoid, 11.5 x 6.9 (10-14 x 6-8) micrometer. Sporulated oocysts of Eimeria cochisensis n. sp. were spheroid to subspheroid, 16.7 x 15.3 (15-18 x 14-17) micrometer, with sporocysts ovoid, 8.4 x 5.6 (6-11 x 4-7) micrometer. Fifty-five of 71 (77%) infected hosts had oocysts of only one eimerian species in their feces at the time they were examined. One eimerian, E. tamiasciuri, was found in seven of nine host species in three genera. A list is provided of all eimerians (22, including the species described here) that have been described in the literature from Eutamias, Sciurus, and Tamiasciurus spp.

Animals↗

Effect of conditioned media from chicken and turkey intestinal cell cultures on invasion by sporozoites of three species of avian coccidia.

The effect of conditioned media from cultures of turkey and chicken intestinal cells on cellular invasion by sporozoites of avian Eimeria species was examined in vitro. Media conditioned by the growth of cells from the ceca, mid-intestine (area of the yolk stalk diverticulum), and duodenal loop were examined for their ability to enhance invasion. Conditioned medium from cultures of turkey cecal cells significantly enhanced invasion by the turkey coccidia Eimeria adenoeides, by 2.4-fold, and E. meleagrimitis, by 2.2-fold, as compared with invasion in the presence of control medium. Conditioned medium from mid-intestinal cell cultures enhanced invasion by the two coccidial species by 2.0- and 2.1-fold, respectively. The enhancement occurred with conditioned media from early (1) as well as later (11) passages of cells. This suggests that the enhancing factor was produced by fibroblast-like cells, the predominant cell type at both early and late passages, and not by epithelial-like cells that had disappeared by the first or second passage. Additionally, conditioned media from cultures of chicken cecal and duodenal loop cells significantly enhanced invasion by the turkey cecal coccidium, E. adenoeides, (1.7- and 1.6-fold, respectively). This was less enhancement than was caused by the turkey cell conditioned media. Heat treatment (56 degrees C for 45 min) of conditioned media failed to alter the effect on invasion. Neither the turkey or chicken cecal cell media nor conditioned media from any other chicken intestinal cell cultures enhanced invasion by E. tenella, the chicken cecal coccidium. Although morphologically dissimilar when they were first plated, the gross appearance and growth of the turkey and chicken cells when conditioned media was collected was comparable.

Animals↗

Control of coccidia in young calves using lasalocid.

Thirty-six, 2- to 4-day-old Friesian bull calves were divided into 4 groups and fed milk replacer and calf starter pellets ad libitum in separate pens. Four treatments were applied; lasalocid in milk (1 mg/kg body weight/day) (M), lasalocid in starter (F), lasalocid in both milk and starter (M + F) and untreated (C). When the calves were about 2 weeks old they were each dosed orally with 550,000 sporulated Eimeria sp oocysts, mainly E zurneii and E bovis. The infection, detected by faecal excretion of oocysts, was suppressed in the M + F and M groups. There was significant excretion of oocysts in the F group but these calves did not show any clinical signs of coccidiosis. Untreated calves were affected with diarrhoea containing blood on the 24th day after inoculation. Body weight gain and intake of starter pellets was also depressed in the untreated calves during the time they were clinically affected. It is concluded that mixing lasalocid in milk replacer (or fresh milk) is an effective method of protecting young calves against early infection with coccidia.

Animals↗

Strain-dependent differences in murine susceptibility to coccidia.

Differences in susceptibility of strains of mice to Eimeria ferrisi were observed by infecting eight strains of mice with six infectious dose levels and comparing the mortality rate among the strains for a period of 12 days. Mice of the C57BL/6 and HA/ICR strains were susceptible, and those of A/He, AKR, BALB/c, CBA, C3H/Anf, and DBA/2 strains were resistant to coccidial infection. Resistance was a dominant genetic expression, as indicated by the resistant response of F(1) hybrids of susceptible C57BL/6 and resistant CBA, C3H/Anf, or DBA/2 strains. An E. ferrisi infection in congenitally athymic nu/nu mice and phenotypically normal heterozygous nu/+ mice was used to determine how thymus-dependent immunoincompetence in cell-mediated immunity of the nu/nu mouse affected resistance to infection in a genetic background of the resistant BALB/c mouse. Results of primary and challenge infections in these two strains of mice suggested that resistance is thymus dependent. Furthermore, impairment of thymus-dependent cell-mediated immunity in resistant AKR mice by treatment with mouse antithymus serum led to partial susceptibility. However, susceptible C57BL/6 and HA/ICR strains are phenotypically normal mice, and previous evidence showed that C57BL/6 mice are not completely immunoincompetent in cell-mediated reactivity to coccidia. Collectively, our data show that cell-mediated immunity is necessary for resistance but may be subjected to modification by genetic expression of the host. The possible role of immune response genes in the control of coccidial immunity is discussed.

Animals↗

Cross-protection against four species of chicken coccidia with a single recombinant antigen.

A cDNA clone, SO7', from an Eimeria tenella cDNA library was inserted into the high-expression vector pJC264 and was expressed in Escherichia coli as a fusion protein, CheY-SO7', with a molecular mass of approximately 36 kDa. By using the purified recombinant antigen to immunize young chicks, it was demonstrated that a single dose, without adjuvant, not only protected against severe coccidiosis induced by infection with E. tenella but also protected chicks challenged with the heterologous species Eimeria acervulina, E. maxima, and E. necatrix. By using rabbit antiserum raised against recombinant CheY-SO7', Western blot (immunoblot) analysis of sporulated oocysts of all seven major species of chicken coccidia showed that all species tested contained proteins characteristic of the B class of antigens, of which CheY-SO7' is representative. It seems likely that a single B antigen could protect chickens against severe coccidiosis caused by infection with any of these Eimeria species. Although chicks exposed to prolonged, natural infection develop antibodies to B antigen, active immunization of young chicks with a protective dose of CheY-SO7' does not elicit a humoral antibody response, suggesting that the partial protection results from cell-mediated effector mechanisms. In addition, the cross-protective nature of the immunity indicates that the response to B antigen is different from that induced by natural infection, which elicits a species-specific immunity. To date, the protection induced by B antigen immunization, although remarkable for a single recombinant protein, is not sufficient to compete with prophylactic chemotherapy.

Animals↗

Intestinal coccidia (Apicomplexa: Eimeriidae) of Brazilian lizards. Eimeria carmelinoi n.sp., from Kentropyx calcarata and Acroeimeria paraensis n.sp. from Cnemidophorus lemniscatus lemniscatus (Lacertilia: Teiidae).

Eimeria carmelinoi n.sp., is described in the teiid lizard Kentropyx calcarata Spix, 1825 from north Brazil. Oocysts subspherical to spherical, averaging 21.25 x 20.15 micro m. Oocyst wall smooth, colourless and devoid of striae or micropyle. No polar body or conspicuous oocystic residuum, but frequently a small number of fine granules in Brownian movement. Sporocysts, averaging 10.1 x 9 microm, are without a Stieda body. Endogenous stages characteristic of the genus: intra-cytoplasmic, within the epithelial cells of the ileum and above the host cell nucleus. A re-description is given of a parasite previously described as Eimeria cnemidophori, in the teiid lizard Cnemidophorus lemniscatus lemniscatus. A study of the endogenous stages in the ileum necessitates renaming this coccidian as Acroeimeria cnemidophori (Carini, 1941) nov.comb., and suggests that Acroeimeria pintoi Lainson & Paperna, 1999 in the teiid Ameiva ameiva is a synonym of A. cnemidophori. A further intestinal coccidian, Acroeimeria paraensis n.sp. is described in C. l. lemniscatus, frequently as a mixed infection with A. cnemidophori. Mature oocysts, averaging 24.4 x 21.8 microm, have a single-layered, smooth, colourless wall with no micropyle or striae. No polar body, but the frequent presence of a small number of fine granules exhibiting Brownian movements. Sporocysts 9 x 8, without a Stieda body. Endogenous stages epicytoplasmic, characteristic of the genus, in the upper ileum. The importance of a study of the endogenous stages of eimeriid coccidia is discussed.

Animals↗

Intestinal coccidia in Cuban pediatric patients with diarrhea.

From May to August 1999, we evaluated 401 patients from a pediatric hospital of Havana City. One group was composed of 113 patients with diarrhea admitted to the Gastroenterology ward and a second consisted of 288 patients without diarrhea, admitted for other reasons, and hospitalized within the same time period. Three stool samples were collected from each child and were examined using three parasitological techniques. When we compared the frequency of parasite species between both groups, we found Cryptosporidium spp. and Cyclospora cayetanensis, only in the group of children with diarrhea (P < 0.01). However, no significant differences were found in the occurrence of the other intestinal parasites (P > 0.05). In addition, in those children infected with Cryptosporidium, the diarrhea had a more prolonged duration (P < 0.01), while those infected with Cyclospora, the abdominal cramps or pain, and acute diarrhea were more frequently detected (P < 0.01). Our results showed that emerging intestinal coccidia are pathogens strongly associated in this group of children with diarrhea.

Animals↗

Use of monoclonal antibodies developed against chicken coccidia (Eimeria) to study invasion and development of Eimeria reichenowi in Florida sandhill cranes (Grus canadensis).

Eimeria gruis and Eimeria reichenowi are common coccidial parasites of a number of species of cranes. Until recently, little was known about either the site for invasion or the dynamics of early development of the crane coccidia because of the difficulty of identifying sporozoites and early developmental stages of these parasites by conventional staining methods. In the present study, monoclonal antibodies (MAbs) elicited against Eimeria spp. of chickens and turkeys were found to cross-react with sporozoites and developmental stages of E. reichenowi in the tissues of Florida sandhill cranes (Grus canadensis). With these Mabs, E. reichenowi sporozoites were found in specimens taken at 6 hr postinoculation (PI) from just proximal to Meckel's diverticulum in the jejunum to the ileocecal juncture. Fewer were found in the ceca and rectum and none in the duodenal loop. At 24 hr PI, there were markedly fewer sporozoites and their location had shifted to the duodenum. No stages were seen in intestinal cells at 5 days PI (DPI), but trophozoites had developed in the liver and spleen. At 10 DPI, sexual stages were detected in the intestine from the duodenal loop through Meckel's diverticulum but not in other organs. By 14 DPI, numerous developmental stages were detected in the intestine (ceca and jejunum), liver, and lungs but not in the heart, kidney, or brain. The number, location, and maturity of the stages in the ceca differed markedly from those in the jejunum.

Animals↗

Battery efficacy studies with arprinocid against field strains of Coccidia.

Graded levels of 9-[2-chloro-6-fluorophenyl)methyl]-9H-purin-6-amine (arprinocid) ranging from 30 ppm to 90 ppm in the diet have been tested for anticoccidial efficacy in controlled battery experiments against 20 field isolates of coccidia. These isolates, representing five major species of Eimeria, were collected from various geographic locations and all but one were found to be resistant to one or more of the currently marketed products. Arprinocid prevented mortality, increased weight gains, and reduced or eliminated intestinal lesions and oocyst production. A level of 60 ppm arprinocid provided adequate control of all isolates and no evidence of cross resistance was observed between arprinocid and the marketed products tested. On the basis of these studies, it is concluded that arprinocid at levels of 60 ppm to 70 ppm is effective in controlling coccidiosis in broilers, even that caused by isolates refractory to many of the marketed products.

Adenine↗

Pathogenicity of coccidia in Japanese quail (Coturnix coturnix japonica).

Coccidia were recovered from a field outbreak in commercially raised Japanese quail from South Carolina. After propagation in unmedicated quail, the culture was identified as a mixture of approximately 65% Eimeria uzura, 33% E. tsunodai, and 2% E. taldykurganica. Several pure cultures of E. uzura were obtained by single oocyst isolation. A micropyle was not present in all oocysts; thus, it is not a reliable taxonomic characteristic for identification of E. uzura. Neither the mixed culture nor the E. uzura isolates were infective for Bobwhite quail, Chukar partridge, pheasants, chickens, or turkeys. Seventeen-day-old quail were inoculated with various doses of sporulated oocysts ranging from 5 x 10(2) to 5 x 10(5) of the mixed culture and 1 x 10(3) to 1 x 10(6) of E. uzura. The rate of weight gain was depressed at 3 or 4 days postinoculation (DPT) with as few as 5 x 10(2) oocysts/quail of the mixed culture. As few as 1 x 10(3) oocysts of E. uzura produced a weight loss. Some individual quail had depressed packed cell volume and plasma pigment compared with levels in the uninoculated controls. Plasma protein was not affected. Young quail (3 days old at inoculation) were more susceptible than 17-day-old quail. Infection did not adversely affect body weights of adult quail, although egg production was reduced. Mortality was seen only with the mixed culture (100 and 8% in 3- and 17-day-old quail given 5 x 10(5) oocysts, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Age Factors↗

Effectiveness of therapeutic anticoccidial drugs against recently isolated coccidia.

Litter samples were collected weekly from 17 broiler houses and coccidial oocysts were counted. Oocysts per gram were low on the Weeks 1 and 2 of the growout, increased to a peak on the Week 4, then declined to near zero on Weeks 6 and 7. Coccidia from each house were propagated and tested for sensitivity to therapeutic formulations of amprolium, sulfaquinoxaline, and sulfaquinoxaline plus pyrimethamine. The degree of control was based on the weight gains and lesion scores of the medicated groups in comparison with those of the unmedicated, infected, and uninfected groups. The predominant species found in the 17 isolates were Eimeria tenella and E. acervulina. Amprolium was effective against E. tenella in all isolates but only partly effective against E. acervulina. Sulfaquinoxaline and the potentiated mixture were effective against the E. acervulina but only partly effective against the E. tenella. The potentiated mixture was better against E. tenella than sulfaquinoxaline alone.

Amprolium↗

Effect of monensin on ultrastructure and cellular invasion by the turkey coccidia Eimeria adenoeides and Eimeria meleagrimitis.

Freshly excysted sporozoites (SZ) of the turkey coccidia Eimeria meleagrimitis and Eimeria adenoeides were incubated at 41 C in concentrations of monensin from .01 to 1.0 microgram/mL, washed free of the drug, and either processed for phase, fluorescence, and transmission electron microscopy or inoculated into cultures of turkey kidney cells. Phase microscopy indicated that after 1.5 h incubation in 1.0 micrograms/mL monensin, about 60% of the SZ of E. meleagrimitis had become notably rounded or displayed localized protrusions. These alterations were accompanied by ultrastructural abnormalities (in 90% of the SZ) including vacuoles in the cytoplasm, bulging and separation of plasma membrane layers, and dense bands in the refractile bodies that extended toward the periphery of the refractile body. Similar morphological and ultrastructural changes were observed in over half of the E. adenoeides SZ after 2 h incubation in 1.0 micrograms/mL monensin. Additionally, some specimens contained a pycnotic nucleus that was usually surrounded by a large vacuole. After 4 h incubation, almost all of the SZ displayed some degree of ultrastructural damage. Indirect fluorescent antibody labeling with parasite-specific monoclonal antibodies demonstrated clouds of antigen surrounding the monensin-treated but not the untreated SZ, suggesting an increase in permeability with incubation in monensin. With both E. meleagrimitis and E. adenoeides, the structural changes were reflected in a significant inhibition of cellular invasion. The inhibitory activity of monensin was concentration- and time-dependent in that the greatest inhibition of invasion was observed in SZ incubated for 4 h in 1.0 micrograms/mL of monensin; shorter incubation times or lower concentrations of monensin having less effect.

Animals↗

Comparative development of Eimeria tenella in primary chick kidney cell cultures derived from coccidia-resistant and -susceptible chickens.

A sixfold difference in resistance to coccidia (Eimeria tenella) infection between a resistant and a susceptible line of Auburn White Leghorn chickens, derived by selective breeding, has been reported. The purpose of the following study was to determine whether the resistance or susceptibility phenomenon in the Auburn lines could be manifested in a homogeneous group of isolated host kidney cells that support E. tenella development in vitro but not normally in vivo. Propagation of the parasite in host cells in vitro eliminates humoral and cellular elements of immunity, and allows the study of host genetic influences at the cellular level. Differences in parasite development were examined between the two lines of cells in vitro after 48 and 96 h of incubation; time periods that reflect initial infection of the host cells by the parasite and the subsequent asexual development. Quantification of differences by liquid scintillation counting was based on parasite-specific incorporation of pyrimidines, specifically [3H]-uracil. The results supported previous findings that overall E. tenella development was significantly greater in the host cells from the susceptible line than in the cells from the resistant cultures at both time periods.

Animals↗