Life cycle of Isospora suis in gnotobiotic and conventionalized piglets

Isospora suis had 3 asexual and 1 sexual intra-intestinal conventional life cycle. The first asexual generation was most prominent at 2 days p.i. (post inoculation) and produced 2–7 merozoites. The second-generation meronts were prevalent at 3–4 days p.i. and produced 2–12 large merozoites. At 4–5 days p.i. the third generation meronts were prominent and produced 4–24 small crescent shaped merozoites. Mature sexual stages were most prominent at 5–6 days p.i. The stages were most numeroous in the distal half of the small intestine. At 8–9 days p.i. stages morphologically similar to the second generation of meronts reappeared, followed by the further development into third generation merozoites and sexual stages. This was reflected in a prepatent period of 5 days and a biphasic patent period of 5–8 or 9, and 11–14 days p.i.Intraperitoneal injection of liver/spleen and intestinal lymph node homogenates, respectively, from piglets infected 24 and 48 h, previously with high doses of oocysts, resulted in a patent infection 10–12 days post inoculation of the donor piglets. No differences in the life cycle of I. suis were observed between conventionalized and germ-free piglets. An extra-intestinal life cycle of I. suis related to the second patent period was postulated.     

Additional details of the life history of the chicken coccidium Eimeria mitis Tyzzer, 1929

Four asexual generations of Eimeria mitis were identified. The first three developed above the epithelial cell nuclei, but the fourth developed above and below. Meronts measured 13.8 x 16.4 microns, 16.1 x 16.4 microns, 12.1 x 14.6 microns, and 9.5 x 12.4 microns, respectively, of generations 1, 2, 3, and 4. They matured at 36, 67, 72, and 88 hr postinoculation (PI) and contain 20-24, 16-20, 10-14, and 7-10 merozoites, respectively. Merozonts measured 7.2 x 1.9 microns, 8.5 x 2.5 microns, 9.6 x 2.0 microns, and 6.75 x 2.75 microns, respectively. The first two types of meronts were deep in the crypts and epithelial cells. The third and fourth types of meronts were along the side and tip of the villi. Gametocytes developed from third and fourth generation. Gamonts were usually below the nuclei of the epithelial cells. Parasitism was primarily in the ileum, ceca, and rectum and also in the yolk-sac diverticulum.     

Development of Isospora suis from pigs in primary porcine and bovine cell cultures

Sporozoites of Isospora suis penetrated and developed by endodyogeny in primary porcine kidney (PPK) and primary fetal bovine kidney (PFBK) cell cultures. Motile merozoites and binucleate Type I meronts were observed in both types of cultured cells. Multinucleate Type II meronts developed in PPK cell cultures only. These multinucleate meronts were always found singly, were nonmotile and did not form merozoites.     

Life cycle, host restriction and longevity of Cyrilia nili (Haemogregarina nili Wenyon, 1909) n. comb.

Morphometric and morphological observations on C. nili in an experimentally infected fish (using the leech vector Batracobdeloides tricarinata for the first time) revealed two successive types of merogonic cycles occurring within the fish erythrocytes. In the first cycle large meronts produced eight small merozoites each while in the second cycle smaller meronts produced four merozoites each. Merozoites of the second cycle were destined to become gamonts. The gamonts were somewhat larger than merogonic stages and comprised the majority of the parasitic stages during parasitemia for up to 8 months. In the leech crop, gamonts released from the fish erythrocytes, became associated in syzygy and fused. The formed zygote underwent sporogony within the intestinal tissues and up to 60 sporozoites were produced from an ovoid or round oocyst. The produced sporozoites migrated toward the salivary and the proboscis tissues. The parasite survived in the leech over a period of 8 months involving 5 meals after the initial meal provided the fasting period did not exceed 75-105 days. Survival was attributed to residual stages in the proboscis and salivary tissues. Cross transmission experiments between fishes via B. tricarinata revealed that the fish haemogregarines were not host specific. Previously described African fresh water fish haemogregarines were indistinguishable and on the basis of the current results, they are regarded as a single species Cyrilia nili.     

Life cycle of Eimeria procera in experimentally infected grey partridges (Perdix perdix)

We examined exogenous and endogenous development of Eimeria procera in experimentally infected grey partridges (Perdix perdix). Our examination included data on morphology, localization, duration of schizogony and gametogony and morphology of sporulated oocysts. The endogenous stages of E. procera developed in large numbers within the epithelial cells of caecal crypts. The asexual development comprised three generations of schizonts. The first fully developed macrogametes and microgamonts were observed on Day 5 post-infection (p.i.) in histologic section. The patent period began on Day 6 p.i. and ended on Day 11 p.i. with peak production of oocysts on Days 7 and 8. Long oval oocysts of E. procera measured 25.78–28.13 μm in length and 14.06–15.24 μm in width, sporulation time ranged from 18 to 24 h at 25°C and from 36 to 48 h at 20°C.     

Immunogenicity of the different stages of Eimeria falciformis var. pragensis

The immunogenicity of the different stages of E. falciformis var. pragensis was evaluated with the indirect fluorescent antibody reaction. Sporozoites, mature asexual stages and merozoites treated with sera from immune mice and with fluorescein-conjugated goat antimouse gammaglobulin showed the most intence specific fluoresence. The immature asexual stages and the sexual stages fluoresced weakly, whereas the sporocysts and oocysts did not specifically fluoresce. On the basis of the intensity of the immunofluorescence, the sporoziotes and mature asexual generations (merozoites) were considered to be more immunogenic than that immature asexual and the sexual stages. Antibodies against sporocysts and oocysts could not be demonstrated.     

Merogony and gametogony of Eimeria mccordocki (Protozoa-Eimeriidae) in the mule deer, Odocoileus h. hemionus

Eimeria mccordocki and E. madisonensis oocysts were isolated from feces of 21 of 40 captive mule deer in Fort Collins, Colorado. The two species were separated from each other by infecting one mule deer fawn, and the life cycle of E. mccordocki was studied for the first time. Four to six-weeks-old mule deer fawns were inoculated orally with E. mccordocki and killed 9, 13 and 15 days after infection. Asexual and sexual stages of life cycle developed in the ileum of mule deer, only in the surface epithelial cells of the villi. The asexual stages consisted of two generations of meronts.     

杀球灵对鸡盲肠球虫病的治疗试验及作用峰期探讨

本试验用１０日龄公雏９６只,分为８个组。氯嗪苯乙氰用量为１×１０－６,其中６个组分别在感染后０,４８,７２,９６,１２０,１５４ｈ给药治疗人工感染１×１０５个柔嫩艾美耳球虫孢子化卵囊的雏鸡。结果表明杀球灵（Ｃｌｉｎａｃｏｘ）为高效抗球虫药物,兼有预防和治疗作用。作用于盲肠球虫的有性生殖和无性生殖阶段。     

Immunohistologic examination of monoclonal antibodies generated against Eimeria bovis sporozoites for reactivity to meronts and sexual stages of E bovis and other eimerian parasites

Seven monoclonal antibodies (MAB) generated against sporozoites of Eimeria bovis were tested for reactivity against immature and mature first-generation meronts, sexual stages, and oocysts in tissues from experimentally infected calves by use of an avidin-biotin peroxidase complex (ABPC) immunohistologic test. Three of the 7 MAB reacted in the ABPC test. One of these, MAB-4FB4, reacted only with mature E bovis meronts. The other 2 MAB, MAB-2AE7 and MAB-4AD7, reacted with all the developmental stages of E bovis tested. Asexual stages and sexual stages of E tenella from chickens and E papillata from mice also were examined in the ABPC test. Monoclonal antibodies MAB-2AE7 and MAB-4AD7 reacted with all stages of these eimerian protozoa. None of the other 5 MAB reacted with these parasites. Results of this study suggested that antigens are shared among the asexual and sexual stages of several diverse Eimeria species.     

Administration of toltrazuril during experimental coccidiosis in mule ducks: comparison of the efficacy of a single administration at two different endogenous stages

An experimental Eimeria mulardi coccidiosis was reproduced in mule ducks. A single dose of toltrazuril (7 mg per kg) was administered at two different endogenous stages: Day 2 (first meronts) or Day 5 (last meronts and gamonts). The efficacy of the drug was assessed on four criteria (body weight, oocyst counts, macroscopic lesions, and presence of the meronts and gamonts). To have a curative effect, a single dose of toltrazuril should be administered early on. However, a single treatment on Day 5 had a prophylactic effect.     

Ultrastructure of developing Isospora suis in cultured cells

The ultrastructure of Isospora suis sporozoites, type-1 meronts, and type-1 merozoites was examined, using transmission electron microscopy of infected cultured cells. The ultrastructure of sporozoites and type-1 merozoites was similar. Each possessed trimembranous pellicles, subpellicular microtubules, a conoid, anterior and posterior polar rings, rhoptries, micronemes, a single vesicular nucleus, tubular mitochondria, Golgi complexes, ribosomes endoplasmic reticula, inactive micropores, amylopectin bodies, lipid bodies, dense bodies, and crystalloid bodies. Merozoites were produced by endodyogeny. Ultrastructural events associated with merozoite production by type-1 meronts are described.     

Porcine neonatal coccidiosis in quebec

Intestinal coccidiosis was diagnosed in 110 diarrheic piglets originating from 66 farrowing operations. These piglets had been submitted alive for necropsy early after the beginning of diarrhea, and the diagnosis of coccidiosis was based on the demonstration of coccidial forms in their jejunum by histopathological examination. Enteropathogenic strains of Escherichia coli were demonstrated in only nine of the 110 piglets while the transmissible gastroenteritis virus and rotavirus were each present in three pigs. The most common lesion was a multifocal villous atrophy in the middle and lower jejunum, and ileum of the affected pigs. Coccidial forms were present in variable numbers of the cells lining the affected villi and they were mainly asexual stages (meronts and merozoites).

In the affected farrowing houses diarrhea began between five and 15 days of age with the highest frequency around seven to ten days of age. Morbidity rates were very variable and mortality was less than 20%. The disease was a persistent problem in large farrowing operations on continuous farrowing programs, and losses were due mainly to retarded growth and treatment costs which were usually inefficient. The disease occurred year round with the highest prevalence in summer and fall months, and in January.

Search for oocysts in fecal samples or colonic contents from 28 of the 110 pigs revealed that 13 of them were shedding oocysts identified as Isospora suis.

     

Congenital sporozoan encephalomyelitis in a calf

Protozoan encephalomyelitis was diagnosed post mortem in a five-day-old Friesian calf which had shown nervous signs from birth. The lesion was a subacute necrotising multifocal encephalomyelitis associated with protozoan bodies (6 X 6 microns to 16 X 30 microns). Ultrastructurally these bodies corresponded to apicomplexan meronts composed of eight to 89 merozoites which reproduced by internal budding to form paired daughter cells (endodyogeny). The merozoites were indistinguishable from Toxoplasma gondii; they reacted weakly with anti-Sarcocystis species serum and did not cross react with anti-T gondii serum. The generic identity of the sporozoan was not established, but it is unlikely to have been either Sarcocystis species or T gondii.     

Studies on synchronous egress of coccidian parasites (<Emphasis Type="Italic">Neospora caninum,Toxoplasma gondii,Eimeria bovis)</Emphasis> from bovine endothelial host cells mediated by calcium ionophore A23187

Neospora caninum, Toxoplasma gondii and Eimeria bovis are coccidian parasites of veterinary importance. Tachyzoites of N. caninum and T. gondii and sporozoites of E. bovis are able to invade and replicate in endothelial cells in vivo and in vitro. As it holds true for all eukaryotic cells, the survival of parasitized host cells and the parasites themselves should be dependent on ion balances, especially on extra- and intracellular calcium concentrations. Addition of the calcium ionophore A23187 reliably did release merozoites from mature N. caninum and T. gondii meronts grown in cultured primary bovine umbilical vein endothelial cells (BUVEC). Extent and time course of merozoite release depended on both, maturity of the meronts and concentration of the calcium ionophore. Attempts, however, to achieve synchronous release of merozoites from E. bovis first generation meronts by ionophore treatment failed, suggesting a different biological behaviour of this parasite. According to microscopical observations, the quite variable time of E. bovis macromeront maturation and a hampered merozoite exit owing to dense parasite-induced cytoskeleton elements surrounding the meront may be a reason for the lack of inducible synchronous release. Electronic supplementary materials The online version of this article (doi: ) contains supplementary material, which is available to authorized users.     

Canine coccidiosis attributed to an Isospora ohioensis-like organism: a case report

Coccidiosis due to an Isospora ohioensis-like organism was diagnosed in a 10-week-old pup. The pup had diarrhea, dehydration, and weight loss before it died 7 days after onset of clinical signs. Lesions were limited to the intestinal tract. Beginning in the distal end of the small intestine and extending through cecum and colon were mild histiocytic proliferation in the lamina propria and multifocal cryptitis. Numerous meronts and gamonts of an unidentified coccidium were in or near the lesions. Parasites were in the villous epithelium, lamina propria, and intestinal glands of the distal one-half of the small intestine, cecum, and colon. It was concluded that infection was due to an unidentified coccidium with oocysts structurally similar to those of I ohioensis.     

The detection of IgM and IgG antibodies against Babesia bigemina in bovine sera using semi-defined antigens in enzyme immunoassays

Soluble extracts prepared from Babesia bigemina merozoites were tested for antigenicity in class-specific enzyme immunoassays currently being evaluated for the differential serodiagnosis of bovine babesiosis. Intact merozoites were harvested from erythrocytes from an experimentally-infected calf by controlled hypotonic lysis and differential ultra-centrifugation. The merozoites were disrupted by ultrasonication and a crude soluble extract obtained by ultracentrifugation. Fractionation of the crude extract on calibrated Sephadex G-200 columns consistently produced 4 fractions with molecular weights of 600, 40, 15 and 5 k (k = 10(3) daltons). Only the 600 and 15 k fractions proved to be antigenic when reacted against bovine immune sera. These fractions were incorporated into IgM- and IgG-specific enzyme immunoassays and used to determine the kinetics of the host-antibody responses to infection. The use of semi-defined antigens allowed assay standardization and good reproducibility of the results. A calf infected with a cryopreserved stabilate of B. bigemina originating from adult Boophilus microplus ticks developed a mild transient fever from 6-4 days post-infection (d.p.i.) and low parasitaemia levels from 7-16 d.p.i. IgG-antibodies first appeared at 7 d.p.i., peaked in intensity at 12 d.p.i. and then persisted at these levels until the end of the test period at 49 d.p.i. IgM-antibodies appeared at 7 d.p.i., peaked in intensity from 12-22 d.p.i., but then declined to low levels by 28 d.p.i. The importance of this transitory IgM-antibody response in the serodiagnosis of acute B. bigemina infections remains to be determined in clinical and field situations.     

Antibody response against endogenous stages of an attenuated strain of Eimeria tenella

The application of attenuated vaccines for the prevention of chicken coccidiosis has increased exponentially in recent years. In Eimeria infections, protective immunity is thought to rely on a strong cell mediated response with antibodies supposedly playing a minor role. However, under certain conditions antibodies seem to be significant in protection. Furthermore, antibodies could be useful for monitoring natural exposure of flocks to Eimeria spp. and for monitoring the infectivity of live vaccines. Our objective was to investigate the chicken antibody response to the different parasite life cycle stages following infection with an attenuated strain of Eimeria tenella. Western blotting analysis of parasite antigens prepared from the lining of caeca infected with the attenuated strain of E. tenella revealed two dominant antigens of 32 and 34 kDa, apparently associated with trophozoites and merozoites that were present at high concentrations between 84 and 132 h post-infection. When cryosections of caeca infected with E. tenella were probed with IgY purified from immune birds the most intense reaction was observed with the asexual stages. Western blotting analysis of proteins of purified sporozoites and third generation merozoites and absorption of stage-specific antibodies from sera suggested that a large proportion of antigens is shared by the two stages. The time-courses of the antibody response to sporozoite and merozoite antigens were similar but varied depending on the inoculation regime and the degree of oocyst recirculation.     

Ultrastructural observations on different developmental stages of Goussia sinensis (Chen, 1955), a parasite of the silver carp (Hypophthalmichthys molitrix Valenciennes, 1844)

The development of Goussia sinensis, a coccidium parasitizing the intestine of the silver carp (Hypophthalmichthys molitrix) was studied by electron microscopy. All stages developed in the epithelial cells, less frequently in the goblet cells, and were located within a parasitophorous vacuole. In some cases one cell was invaded by several merozoites. Eight to sixteen merozoites were formed within the meront by ectomerogony. The ultrastructural processes characteristic of gamogony were the same as those found for Goussia spp. parasitizing other species of fish. A hitherto unknown mechanism of oocyst wall formation was observed. The oocyst membrane developing within the zygote surrounded only part of the zygote material. Thus, a small part of the zygote material left the oocyst proper. It is suggested that this zygote residue and the necrotic host cell constitute the so-called "yellow bodies" which include the excreted oocyst. The oocyst wall was 40 to 60 nm thick. Oocyst sporulation took place within the fish. The sporocysts consisted of two hemispheres connected by sutures and had a 100 to 120 nm thick double wall. They were surrounded by sporocyst veils fixed to the oocyst wall by membranes.     

Cryptosporidium parvum: investigation of sporozoite excystation in vivo and the association of merozoites with intestinal mucus

Enteric cryptosporidiosis was studied in the small intestine of five-day-old sucking mice after infection with 10(6) Cryptosporidium parvum oocysts. It was shown that excystation and the majority of subsequent endogenous stages occurred predominantly in the ileum. During the first three days of infection the number of merozoites collected in ileal washings increased over 100-fold to approximately 10(6) merozoites per mouse on the third day. In contrast to control mice, wash fluid from infected mice contained numerous strands of dislodged mucus. Estimates of mucus in the ileal washings of infected mice were similar to those made in controls until day 4 after infection when they increased and remained high throughout the remainder of the experiment. This study describes a method whereby ileal mucus washings from C parvum infected infant mice could be used as a rich source of merozoites.