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An age-dependent aspect of resistance to Cryptosporidium muris (strain MCR) infection was monitored in Syrian golden hamsters, Mesocricetus auratus, at 1-, 5- and 10-week of age and in ICR mice. Mus musculus, at 3-, 12-, and 15-week of age orally inoculated with a single dose of 2 x 10(6) oocysts, respectively. The prepatent periods for both animals were similar, independent of age, but the patency was significantly longer in younger hamsters (P < 0.001) and a long tendency in younger mice. Hamsters infected at 1-week of age excreted about 10 times higher oocysts than those at 5- and 10-week of age. However, the total oocyst output was similar among mice of different ages. There was a good correlation between the length of the patency and the total oocyst output in hamsters (R = 0.9646), but not in mice (R = 0.4561). The immunogenicity of the parasite to homologous challenge infections was very strong in hamsters and relatively strong in mice. These results indicate that acquired resistance to C. muris infection is age-related and the innate resistance is independent of age of hamsters, and that both innate and acquired resistance, on the contrary, are irrespective of age of mice.

Two-day-old commercial chicks were inoculated orally with 2 x 10(6) oocysts of Cryptosporidium baileyi and vaccinated with 10(3.5) EID50/head of a commercially available avian infectious bronchitis (IB) live virus vaccine at 4 and 14 days following inoculation. Chicks infected with C. baileyi were shown to have an immunosuppressive effect on IB virus. It is concluded that infection with the protozoon in early life may increase their susceptibility to IB.

A total of nine Korean native kids and two Corriedale lambs, 1-20 days old, were each inoculated per os with a single dose of 2 x 10(7) oocysts of Cryptosporidium muris (strain MCR) originated from mice to elucidate the kinetics and developmental stages of the coccidium in small ruminants. Irrespective of host's age, the prepatent period for both animals ranged from 19 to 35 days (28.1 days, on the average) and the patent period 16-85 days (47.8 days), and the total oocyst outputs showed enormous differences. Infection with greater numbers of oocyst outputs was not ordinarily established by transmission experiments. Oocysts discharged from the kids retained their infectivity by the mouse titration method. The immunogenicity of the coccidium and oocyst reproduction were proven by challenge infection and administration of prednisolone acetate, respectively. All the developmental stages of the coccidium in parasitophorous vacuoles were found by transmission electron microscopy in the pits of the gastric glands of a kid inoculated with oocysts and then necropsied on day 44 postinoculation. It indicated the full course of the host-parasite relationship in kids and lambs as well as mice.

Hemagglutination-inhibition titers (log2) to Newcastle disease (ND) virus were chronologically observed in chicks, which were orally inoculated with 5 x 10(5) oocysts of Cryptosporidium baileyi at 2 days of age and subsequently vaccinated with inactivated ND virus at 4 and 21 days postinoculation. In general, the titers were considerably lower in the infected chicks than those in the uninfected control throughout the experimental period (p < 0.01), and rapid negative seroconversions were observed in the infected chicks. The titers reached a peak on weeks 2 and 4 post-booster-vaccination in the control and infected chicks, respectively. Thus, C. baileyi infection was shown to have an immunosuppressive effect on ND vaccination when the agent was given to 2-day-old chicks. It is suggested that C. baileyi infection in chicks may increase the host susceptibility to ND virus.

Hemagglutinin (HA) titers to sRBC were chronologically observed in chickens orally inoculated at 2 days of age with 5 x 10(5) oocysts of Cryptosporidium baileyi. All the infected chickens exhibited negligible HA titers by 44 days postinoculation (PI). The titers were elevated as time progressed, and peaked on day 52 PI, declined gradually thereafter, and eventually reached to normal titers on day 92 PI. On the contrary, the titers in uninfected chickens were higher in comparison with infected chickens during the experiment. Chickens infected with the protozoa showed normal oocyst shedding profiles during this period. These data suggest that C. baileyi infection suppress development of humoral immunity to sRBC in chickens. It is possible that impairment of the bursa of Fabricius by cryptosporidiosis rendered chickens vulnerable to other pathogens.

Eight 2-day-old SPF chickens were each inoculated orally with a single dose of 5 x 10(5) oocysts of Cryptosporidium baileyi, and immunoglobulin G (IgG) antibody responses were chronologically measured by indirect immunofluorescent antibody (IFA) assay. Anti-C. baileyi IgG antibody levels remained high (1:106.67 to 1:512.00) for at least 4 months with 330 days of a detectable period. Ten days after the negative conversion, each chicken was re-challenged with 1 x 10(7) oocysts of the same species. Subsequent infection in 340-day-old individuals caused sudden elevated IgG antibody levels and the titer peaked on day 28 postchallenge inoculation (PCI), at 1:1.024 with a 65 days of detection period. Chickens in primary infection showed oocyst shedding profiles, but did not exhibit any oocyst shedding before or after experimental reinfection.

We have successfully maintained Cryptosporidium muris by cryopreservation. Oocysts were suspended in distilled water, stored at -20 degrees C for 24 hrs, and then cryopreserved at -70 degrees C. Cryopreserved specimens were slowly thawed at 5 degrees C. Oocysts, which had been cryopreserved for 15 months without cryoprotective agents, retained their infectivity by the mouse titration method. Oocysts stored at 5 degrees C in 2.5% potassium dichromate failed to retain their infectivity beyond 6.5 months.

Three-week-old ICR SPF mice were orally inoculated with one of 5 doses ranging from 2 x 10(2) to 2 x 10(6) oocysts of Cryptosporidium muris (strain MCR) per mouse. Oocyst inoculation was directly proportional to the amount of oocysts shed and was inversely proportional to the period required for peak oocyst production and to the prepatent period. Peak oocyst production occurred between fifteen and thirty-one days with a patent period from 61 to 64 days. Three days after all mice stopped shedding oocysts, they were orally challenged with a single dose of 2 x 10(6) oocysts of the same species. Marked seroconversion for IgG antibody accompanied recovery from mice inoculated with 5 x 10(5) oocysts. Mice administered with carrageenan excreted a small number of oocysts for 49.0 days on the average after challenge inoculation (ACI) and control mice for 14.2 days in a dose-independent fashion. Just before challenge infection, phagocytic activity of peritoneal macrophages (M phi) and the number of peripheral M phi were dramatically decreased. Mild challenge infection implies that the immunogenicity of C. muris (strain MCR) is very strong, despite M phi blocker carrageenan administration.

Two-day-old chickens and mallards were orally inoculated with one of 5 doses varying from 2 x 10(2) to 2 x 10(6) of C. baileyi oocysts per individual. Generally, the more oocysts inoculated were, the longer the patent periods were, and the more oocysts shedding were. Meanwhile increasing the inoculative dose, the prepatent periods were shortened except that mallards inoculated with 2 x 10(2) and 2 x 10(3) oocysts failed to produce the oocysts. The more parasites involving oocysts appeared from the chicken in comparison to the mallard. In the chickens challenged with a single dose of 2 x 10(6) oocysts, a small number of oocysts were detected from feces on days 4-14 after challenge infection (ACI) in all of carrageenan administered groups and in the control groups inoculated with 2 x 10(2) and 2 x 10(3) oocysts. In the mallards, a few oocysts were also recognized on days 5-15 ACI in all of carrageenan treated groups and in the control groups inoculated with 2 x 10(2), 2 x 10(3) and 2 x 10(4) oocysts. Just prior to challenge infection, phagocytic activity of peritoneal macrophages (Mø) and the number of peripheral Mø in both birds were significantly decreased in the carrageenan treated groups as compared to the control groups. Mild challenge infection in both birds denoted that the immunogenicity of C. baileyi to the birds was very strong, despite Mø blocker carrageenan administration.

Each of SPF chicken (Hi-Line strain, 2-day-old males) was inoculated with 2.5 or 5 x 10(4) oocysts by stomach tube. The oocyst was the medium type of Cryptosporidium previously isolated from Korean chicken origin, and passed in 2-day-old SPF chicken. The patterns of oocyst discharge were monitored daily, and in order to observe the ultrastructure of the developmental stages, the bursa of Fabricius of the chicken was examined by transmission electron microscopy (TEM) on the 12th day postinoculation. The prepatent period for 8 chicken was 5.9 days postinoculation on the average, and the patent period was 12.9 days. The number of oocysts discharged per day for the chicken was reached peak on day 12 postinoculation on the average. A large number of oocysts was found in fecal samples obtained from inoculated chicken on days 8-14 postinoculation. The ultrastructural feature of almost every developmental stage of the medium type from chicken was very similar to that of Cryptosporidium previously isolated from mammalia including human and birds except for the attachment site of C. muris to the mucus cell from mammalia, but dimension of the oocysts from fecal samples of the medium type was different from those of C. meleagridis and mammalia origin. The above results reveal that the medium type of Cryptosporidium of Korean chicken origin is identified as Cryptosporidium baileyi.

Each of SPF mice(Scl: ICR strain, 3-week-old males) was inoculated with 5 x 10(4) oocysts of Cryptosporidium by stomach tube. The oocysts were large type one which was previously isolated from Korean mice, and passaged in 3-week-old SPF mice. The patterns of oocyst discharge were monitored daily, and in order to observe the ultrastructure of developmental stages the stomach of the mice was examined by transmission electron microscopy (TEM) at 4 weeks post-inoculation. The prepatent period for 6 mice was 5.6 days post-inoculation on the average, and the patent period was 63.2 days. The number of oocysts discharged per day from the mice reached peak on day 36.6 post-inoculation on the average. A large number of oocysts were found in fecal samples obtained from inoculated mice on days 30-50 post-inoculation. C. muris was larger than C. parvum at almost every developmental stages, the size difference being 1.4 times in oocysts, 2.4 times in sporozoites, 1.6 times in merozoites, and 1.5 times in microgametes. The ultrastructural features of the attachment site of C. muris to the mucus cells were remarkably different from those of C. parvum and its closely related species. The anterior projection of the protozoa (C. muris), the outer aspect of which was surrounded by a thick filamentous process of the host cell, has not been reported at any developmental stages of C. parvum or its closely related species. The size of the oocysts of strain RN 66 was larger than that of Korean mice origin. The above results reveal that the large type Cryptosporidium of Korean mice origin is identified as Cryptosporidium muris and this type was named as C. muris (strain MCR).

Cryptosporidium, a coccidian protozoa, commonly causes a self-limiting diarrheal illness in humans and animals. Fecal samples from various animals in Chonbuk district were observed using Sheather's flotation technique, Kinyoun's modified acid-fast staining, and osmic acid pre-fixed Giemsa staining. The oocysts were detected in 74 cages (29.6%) out of 250 cages of mature mice, 26 (13.3%) out of 195 mature house rats, 75(15.0%) out of 4-week-old 500 fowls, 98(19.9%) out of 6 to 8-month-old 500 pigs, and 111(22.2%) out of 2 to 5-year-old 500 dairy cattle, respectively. The degree of prevalence was slight in general, but actual prevalence was higher than infection rate because the detection rates were higher in repeated-preparation examinations in comparison to the first examination. Meanwhile, large and small types of oocysts were detected from mice, house rats, pigs, and cattle, and medium type from fowls.