• Parasites, Hosts and Diseases
• /
• v.32 no.1
• /
• pp.7-12
• /
• 1994

A calf and 50 mice were infected with Cryptosporidium parvum and their fecal materials were collected and treated 10 ether extraction (EE), followed by discontinuous sucrose gradients (DSG) method. EE method was to remove some of fat or lipid from feces. Sediments were washed by centrifugation ($1,500{\;}{\times}{\;}g$ for 10 min., 3 times) In phosphate-buffered saline and then these washed sediments were sleeved sequentially through stainless steel screens with a final mesh of 250 ($61{\;}{\mu\textrm{m}}$ porosity) to remove other debris. After sieving, the materials were suspended in 2.5% potassium dlchromate solution. Oocysts were counted by using a hemocytometer and the recovery rate of pure oocysts was calculated on the basis of the count. Following centrifugation ($1,500{\;}{\times}{\;}g$ for 30 min.) by DSG method, most oocysts were recovered at the interface between a gravity of 1.103 and 1.064. The recovery rates of pure oocysts from the fecal suspension of the calf ($3.8{\;}{\times}{\;}10^7/ml$) and the mouse ($3.2{\;}{\times}{\;}10^6/ml$) treated with EE method were 81.6% and 51.6%, respectively. It is suggested that the recovery rate was dependent on the number of oocysts In each suspension treated with EE method. To get the 50% recovery rate, there must be more than $2{\;}{\times}{\;}10^6$ oocysts per ml of the fecal suspension treated with EE method. By the combination of the two methods it was possible to isolate C. parvum oocysts from normal feces of the calf and mouse as well as from dlarrhelc feces.

• Parasites, Hosts and Diseases
• /
• v.33 no.1
• /
• pp.45-54
• /
• 1995

Two-day-old chickens{\;}and{\;}mallards were orally inoculated with one of % doses varying from $2{\;}{\times}{\;}10^2{\;}to{\;}2{\;}{\times}{\;}10^6$ of C. bailevi 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 $2{\;}{\times}{\;}10^2and{\;}2{\;}{\times}{\;}10^3$ oocysts foiled 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 $2{\;}{\times}{\;}10^6$ 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 $2{\;}{\times}{\;}10^2{\;}and{\;}2{\;}{\times}{\;}10^3$ 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 $2{\;}{\times}{\;}10^2,{\;}2{\;}{\times}{\;}10^3{\;}and{\;}2{\;}{\times}{\;}10^4$ oocysts. Just prior to challenge infection, phagocytic activity of peritoneal macrophages (Me) and the number of peripheral Mc in both birds were significantly decreased in the carrageenan treated groups as compared to the control groups. Mild challenge inection in both birds denoted that the immunogenicity of C. bailelli to the birds was very strong, despite MB blocker carrageenan administration.

• Parasites, Hosts and Diseases
• /
• v.29 no.2
• /
• pp.149-160
• /
• 1991

Each of SPF mice(Scl: ICR strain, 3-week-old males) was inoculated with 5$\times$104 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. tsuris was larger than C. parvum at almost every developmental stages, the sixte 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. tsuris 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).

• Parasites, Hosts and Diseases
• /
• v.47 no.3
• /
• pp.293-297
• /
• 2009

Improved methods for detection of Cryptosporidium oocysts in environmental and clinical samples are urgently needed to improve detection of cryptosporidiosis. We compared the sensitivity of 7 PCR primer sets for detection of Cryptosporidium parvum. Each target gene was amplified by PCR or nested PCR with serially diluted DNA extracted from purified C. parvum oocysts. The target genes included Cryptosporidium oocyst wall protein (COWP), small subunit ribosomal RNA (SSU rRNA), and random amplified polymorphic DNA. The detection limit of the PCR method ranged from $10^3$ to $10^4$ oocysts, and the nested PCR method was able to detect $10^0$ to $10^2$ oocysts. A second-round amplification of target genes showed that the nested primer set specific for the COWP gene proved to be the most sensitive one compared to the other primer sets tested in this study and would therefore be useful for the detection of C. parvum.

• Korean Journal of Veterinary Service
• /
• v.30 no.2
• /
• pp.269-274
• /
• 2007

The aim of this study was to examine the epidemiological features of Eimeria in calves with acute diarrhea. Samples were collected from between 15 days and 90 days old calves (n=83) in Gimje area from March 2006 to March 2007. Feces of bloody diarrhea were examined for the presence of Eimeria oocysts using a sucrose flotation method. Out of 83 calves, 62 (74.6%) had Eimeria oocysts. In the results of monthly analysis, the highest prevalence (12.0%) of Eimeria oocysts was found on June. In the seasonal infection rate, spring was the highest prevalence (30.1%), followed by summer (24.0%). Furthermore, the highest prevalence (44.5%) was found in calves from between 31 - 60 days old in the analysis by ages. However, there was no significant differences between female and male sex even though the prevalence was slightly bigger in female than in male. The prevalence of the present study to detect Eimria oocysts for infection may have been affected by weather-conditions in the spring. Young calves should be separated to minimize the infection from cattle as much as possible. Additional studies are necessary to find other factors for infection and combining molecular methods with a highly sensitive system for Eimeria detection could be a reliable and economic way of Eimeria eradication.

• Parasites, Hosts and Diseases
• /
• v.51 no.4
• /
• pp.461-466
• /
• 2013

From May to June 2012, a waterborne outbreak of 124 cases of cryptosporidiosis occurred in the plumbing system of an older high-rise apartment complex in Seoul, Republic of Korea. The residents of this apartment complex had symptoms of watery diarrhea and vomiting. Tap water samples in the apartment complex and its adjacent buildings were collected and tested for 57 parameters under the Korean Drinking Water Standards and for additional 11 microbiological parameters. The microbiological parameters included total colony counts, Clostridium perfringens, Enterococcus, fecal streptococcus, Salmonella, Shigella, Pseudomonas aeruginosa, Cryptosporidium oocysts, Giardia cysts, total culturable virus, and Norovirus. While the tap water samples of the adjacent buildings complied with the Korean Drinking Water Standards for all parameters, fecal bacteria and Cryptosporidium oocysts were detected in the tap water samples of the outbreak apartment complex. It turned out that the agent of the disease was Cryptosporidium parvum. The drinking water was polluted with sewage from a septic tank in the apartment complex. To remove C. parvum oocysts, we conducted physical processes of cleaning the water storage tanks, flushing the indoor pipes, and replacing old pipes with new ones. Finally we restored the clean drinking water to the apartment complex after identification of no oocysts.

• Korean Journal of Veterinary Research
• /
• v.31 no.4
• /
• pp.501-507
• /
• 1991

Diagnosis of cryptosporidiosis is currently confirmed by the detection of the oocysts or endogenous stages in fecal or tissue samples. Various conventional staining methods and serodiagnostic techniques have been reported, but the latter has far been limited to a few laboratories. Cryptosporidium has recently been reported in mice and chiekens in Korea, but there has been no report on staining methods to the oocysts. The present study was performed by light and scanning electron microscopic observations, and discussed with staining properties of four conventional methods such as dichromate solution floatation method, Carbol fuchsin stain, Auramine-O stain and Giemsa stain method. Cryptosporidial oocysts were isolated from the laboratory mouse. In tissue sections of duodenum, jejunum, ileum, cecum and upper colon, numerous very small, basophilic bodies were observed on the border of mucosal epithelial cells. In scanning electron microscopic observations, a few of developmental stages of Cryptosporidium were seen. Two types of thick and thin-walled oocysts were recognized in the intestinal contents. Mean size of its were $5.19{\pm}0.23{\times}4.31{\pm}0.32{\mu}m$ and $5.14{\pm}0.25{\times}4.27{\pm}0.4{\mu}m$, respectively. Carbol fuchsin and Auramine-O stain methods are recommended as the satisfactory ones for the identification of Cryptosporidium oocysts. Giemsa stain was also recommended as available in the laboratory, because a few of developmental stage fo Cryptosporidium could be seen by it.

• Korean Journal of Veterinary Research
• /
• v.30 no.1
• /
• pp.103-106
• /
• 1990

Effects of ammonia water on the spornlation of coccidial oocysts collected from bovine feces were studied with particular reference to the various levels of ammonia water (1% to 10%) for 30 minute conservation at room temperature. The sporulation rates showed a negative linear coorelation according to the treatment leavels of ammonia water, 85.3% at 1% level to 8.9% at 10% level. The optimum level of ammonia concentration was regarded as 5% to 10% with more than 80% of sporulation inhibition effect.

• Journal of Microbiology and Biotechnology
• /
• v.7 no.3
• /
• pp.189-193
• /
• 1997

Cryptosporidium parvum has been recognized as a significant cause of life-threatening diarrhea in Acquired ImmunoDeficiency Syndrome (AIDS) patients. Clinical diagnosis of cryptosporidial infections has been primarily based on the detection of infective stage, oocysts, in stools. Anti-Cryptosporidium oocyst monoclonal antibody (mAb), IgG2a, recognizing an antigen of 97 kDa was generated to be used for diagnosis of Cryptosporidium infection in AIDS patients using an immunofluorecence. It appeared to react with the surface antigens. Transmission electron micrographs of the infective stage of Cryptosporidium recognized by this mAb demonstrated sporolulated oocysts, which measure $4~6{\mu}m$, and sporozoites excysting from oocysts.

• Korean Journal of Veterinary Service
• /
• v.18 no.2
• /
• pp.152-157
• /
• 1995

A total of 1050 layer and broiler chickens from 63 flocks of 21 poultry farms in South-Kyeonggi area, aged from 1 to 18 weeks, were investigated for the prevalence of Cryptosporidium baileyi infection from May 1992 to March 1993. The results have shown that 44 chickens(4.19%) were infected with C.(baileyi during the period of investigation) Fecal samples were treated by using Sheather's flotating technique and were examined under the light and phase contrast microscopy and then stained by Kinyoun method. Cryptosporidium oocysts were found in 35 out of 650 broilers(5.4%) and 9 out of 400 layer chickens(2.3%) aged mainly from 2 to 12 weeks. The regional infection rate were 4.7% in Pyeong-taek, 5.1% in An-sung and 2.3% in Yong-in, respectively. The average size of isolated oocyst was about $5.23{\times}4.92{\mu}m$ and the oocysts were orally inoculated into 2-day-old SPF chickens for the histological examination of oocyst in bursa of Fabricius. The study has concluded that C. baileyi infects chickens and oocysts were isolated in South-Kyeonggi area.