• Korean Journal of Veterinary Research
• /
• v.33 no.4
• /
• pp.649-655
• /
• 1993

A series of experiments on the effects of ${\gamma}-irradiation$ was performed to reveal the pathogenicity of ${\gamma}-irradiated$ oocysts of E tenella from Cobalt-60 and its progeny. The SPF chickens were inoculated with differnt doses of radiation and inoculum. The level of 100 Gy ${\gamma}-irradiation$ from $^{60}Co$ and the level of inoculum with $1{\times}10^4$ oocysts were recognized more pathogenic than those of the other groups by comparison of body weight gains, blood in feces and lesion scores. The signs of blood in feces, lesion score and the number of excreted oocysts in the feces were revealed as the lowest in the group of the ${\gamma}-irradiated$ oocysts, the average in the group of the 1st and the 3rd progeny, and the highest in the group of non-irradiated oocysts of E $tenell\grave{a}$. The body weight gain of the group immunized with ${\gamma}-irradiated$ oocysts of E tenella was higher than those of the non-irradiated, the 1st and 3rd progeny groups. The body weight gain of the groups immunized with the 1st and the 3rd progeny of E tenella were higher than that of the non-irradiated group. The feed conversion ration of the group immunized with ${\gamma}-irradiated$ oocysts of E tenella was lower than those of the non-irradiated, the 1st and the 3rd progeny groups. The feed conversion ratios of the group immunized with the 1st and 3rd progeny of ${\gamma}-irradiated$ oocysts were lower than that of the group infected with non-irradiated E tenella. The anticoccidial index(ACI 190.6) in the chickens immunized with the ${\gamma}-irradiated$ oocysts of E tenella and those(ACI 142.8 and 107.4) of the 1st and the 3rd progeny groups were higher than that (ACT 87.4) of the group infected with non-irradiated E tenella. It was thought that the pathogenicity of ${\gamma}-irradiated$ E tenella would be recovered according to increase the number of generation passaged in chicken.

• Parasites, Hosts and Diseases
• /
• v.33 no.4
• /
• pp.377-382
• /
• 1995

Three-week-old ICR SPF mice were orally inoculated with one of 5 doses ranging from $2{\;}\times{\;}10^2{\;}to{\;}2{\;}\times{\;}10^6$ oocysts of Crwptosporidium tsuris (strain MCR) per mouse. Oocyst inoculation was directly proportional to the amount of oocysts shed and was inversely proportional to the period required for peals 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 $2{\;}\times{\;}10^6$ oocysts or the same species. Marked seroconversion for IgG antibody accompanied recovery from mice inoculated with $5{\;}\times{\;}10^5$ 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. nuris (strain MCR) is very strong, despite $M{\phi}$ blocker carrageenan administration.

• Parasites, Hosts and Diseases
• /
• v.34 no.2
• /
• pp.121-126
• /
• 1996

For the detection of Cwptospori,mum oocysts, fecal samples were collected from 201 calves which showed diarrhea. Among the 201 samples, 29 samples (14.4%) were positive for Cwptosporinium spry. by the DMSO-modified acid-fast stain (MAFS) , 23 samples (11.4%) were positive by commercial kit (Meridian Diagnostics, Cincinnati, Ohiol and 23 by the indirect immunofluorescence antibody (IFA )assay employing the monoclonal antibody (mAb C6). When tested by both IFA and MAFS, 20 fecal samples were positive for Cwptosporinium oocysts whereas 169 fecal samples were negative. If the MAFS is considered a standard method for oocyst detection, the IFA showed 69% of sensitivity and 98% of specificity. When tested by both IFA and commercial kit, 22 fecal samples were positive for Cwptospori,mum oocysts while 177 samples were negative. One sample tested by IFA was found to be false negative, when compared with the results by commercial kit. The sensitivity of IFA was calculated as high as 96%; the specificity as 99% and the predictive value was also 99%. In the present study, IFA employing the nAb C6 revealed that 23 samples (11.4%) were positive among the 201 calves showing diarrhea. Of 23 IFA positive samples, 4 samples (5%) showed cryptosporidial oocysts more than 105 OPG Therefore. it is concluded that the calves showing cryptosporidial oocysts more than 105 OPG in the feces were highly associated with clinical cryptosporidiosis.

• Korean Journal of Veterinary Research
• /
• v.35 no.1
• /
• pp.149-158
• /
• 1995

The purpose of the present study was to understand the pathogenesis of infections in piglets inoculated with C parvum isolated from mice alone and combined with porcine rotavirus (S-80). Thirteen 10-day piglets were divided in four groups; Three, A group, were only given by C parvum. Four, B group, were orally administrated with firstly porcine rotavirus and then C patvum. Three, C group, were orally inoculated with porcine rotavirus alone. The rest, D group, were used as controls. During the experiment, there were daily recorded clinical signs including diarrhea to each pig. According to the periodic intervals for necropsy, all pigs were sacrificed from 4 to 12 days after the final inoculation of C parvum. Location and distribution of two pathogens, C parvum and rotavirus, in the intestinal mucosa of piglets tested were examined by pathological and immunohistological means. In addition, parasitological test using the feces of piglets was applied for the detection of cryptosporidial oocysts as well. A group showed diarrhea from 4 to 6 days post-inoculation(PI) and also discharged C parvum oocysts in feces during the day 4 to 7 PI. In tissue sections of jejunum and ileum, cryptosporidial oocysts were observed a few on the top of villi with slightly fusion. B group represented sign of diarrhea and discharge of oocysts from 2 to 11 days PI. There were some cryptosporidial oocysts both in the jejunal lumen and in the lumen of mucosal glands. As progressed, oocysts were most commonly distributed on the tip of villi of jejunum. Histopathologically there were also mild to moderately fused, attenuated focal desquamated, congested villi and mononuclear cell infiltration of varying degrees in the lamina propria of small intestine and colon at the day 4 and 7 PI. C group showed slightly to mildly attenuated and fused top of villi and mildly mucosal congestion. D group as controls was grossly and histopathologically normal in all parts of intestine. The present results indicate that the piglets inoculated with C parvum only are certainly milder in pathogenesis including duration of clinical course and severity of lesion than those in piglets concurrently infected with porcine rotavirus and C parvum. Also the strain (VRI-CN91) of C parvum used in the study has very low pathogenicity to occur enteritis of piglets.

• Parasites, Hosts and Diseases
• /
• v.56 no.2
• /
• pp.205-210
• /
• 2018

Waterborne parasitic protozoa, particularly Giardia lamblia and Cryptosporidium spp., are common causes of diarrhea and gastroenteritis worldwide. The most frequently identified source of infestation is water, and exposure involves either drinking water or recreation in swimming pools or natural bodies of water. In practice, studies on Cryptosporidium oocysts and Giardia cysts in surface water are challenging owing to the low concentrations of these microorganisms because of dilution. In this study, a 3-year monitoring of Cryptosporidium parvum, Giardia lamblia, and Naegleria fowleri was conducted from August 2014 to June 2016 at 5 surface water sites including 2 lakes, 1 river, and 2 water intake plants. A total of 50 water samples of 40 L were examined. Cryptosporidium oocysts were detected in 22% of samples and Giardia cysts in 32%. Water at the 5 sampling sites was all contaminated with Cryptosporidium oocysts (0-36/L), Giardia cysts (0-39/L), or both. The geometric mean concentrations of Cryptosporidium and Giardia were 1.14 oocysts/L and 4.62 cysts/L, respectively. Thus, effective monitoring plans must take into account the spatial and temporal parameters of contamination because they affect the prevalence and distribution of these protozoan cysts in local water resources.

• Parasites, Hosts and Diseases
• /
• v.52 no.3
• /
• pp.263-271
• /
• 2014

PCR detection of intestinal protozoa is often restrained by a poor DNA recovery or by inhibitors present in feces. The need for an extraction protocol that can overcome these obstacles is therefore clear. $QIAamp^{(R)}$ DNA Stool Mini Kit (Qiagen) was evaluated for its ability to recover DNA from oocysts/cysts directly from feces. Twenty-five Giardia-positive, 15 Cryptosporidium-positive, 15 Entamoeba histolytica-positive, and 45 protozoa-free samples were processed as control by microscopy and immunoassay tests. DNA extracts were amplified using 3 sets of published primers. Following the manufacturer's protocol, the kit showed sensitivity and specificity of 100% towards Giardia and Entamoeba. However, for Cryptosporidium, the sensitivity and specificity were 60% (9/15) and 100%, respectively. A series of optimization experiments involving various steps of the kit's protocol were conducted using Cryptosporidium-positive samples. The best DNA recoveries were gained by raising the lysis temperature to the boiling point for 10 min and the incubation time of the InhibitEX tablet to 5 min. Also, using a pre-cooled ethanol for nucleic acid precipitation and small elution volume ($50-100{\mu}l$) were valuable. The sensitivity of the amended protocol to Cryptosporidium was raised to 100%. Cryptosporidium DNA was successfully amplified by either the first or the second primer set. When applied on parasite-free feces spiked with variable oocysts/cysts counts, ${\approx}2$ oocysts/cysts were theoretically enough for detection by PCR. To conclude, the Qiagen kit with the amended protocol was proved to be suitable for protozoan DNA extraction directly from feces and support PCR diagnosis.

• Parasites, Hosts and Diseases
• /
• v.54 no.3
• /
• pp.339-344
• /
• 2016

The genus Sarcocystis is not usually considered as an important enteric pathogen in immune compromised patients. It might be expected that species for which humans are the final host (Sarcocystis hominis and Sarcocystis suihominis as well as possibly others) would be encountered increasingly often in immunodeficient persons. This study aimed to address how to detect and differentiate Sarcocystis oocysts and/or sporocysts from enteric protozoans in the diarrheal samples of immunodeficient patients in Shiraz, Iran. Diarrheal samples of 741 immunodeficient patients with recurrent persistent or chronic diarrhea were examined by microscopy and molecular biological analysis. Oocysts-positive samples were 68 Cryptosporidium spp., 9 Cystoisospora belli (syn. Isospora belli), 2 Cyclospora cayetanensis, and 15 microsporidia (Enterocytozoon bieneusi). Sarcocystis-like sporocysts found from a woman were identified as Sarcocystis cruzi through 18S rDNA amplification and phylogenetic analysis. To the best of our knowledge, this is the first report of S. cruzi from a human.

• Parasites, Hosts and Diseases
• /
• v.55 no.2
• /
• pp.129-135
• /
• 2017

A total of 60 samples were collected from 35 swimming pools in Beijing, China, and the presence of Cryptosporidium and Giardia were investigated. The results showed that 16.7% and 15.0% of samples were positive for Cryptosporidium oocyst and Giardia cysts, respectively, with a mean concentration of 0.30 oocysts/10 L and 0.27 cysts/10 L. The oocysts and cysts were found to have higher rates of occurrence in August than in May. Genotyping confirmed the presence of Cryptosporidium hominis, C. parvum, and Giardia assemblages A and B, all of which were associated with human infections. The predominant species/assemblages were C. hominis and Giardia assemblage A. Analyses of the relationships between parasite oocysts/cysts, indicator bacteria, and physical-chemical parameters revealed that there was no correlation between 2 parasites and fecal bacterial indicators, whilst there was a significant correlation between protozoa and urea concentration, which indicates that urea concentration rather than fecal bacterial indicators might be an appropriate index for chlorine-resistant protozoa in swimming pools. This study provides useful information to improve the safety of swimming pool water and deduce the risk of protozoan infections.

• Korean Journal of Veterinary Research
• /
• v.51 no.3
• /
• pp.249-252
• /
• 2011

Cell culture systems for the protozoan Eimeria are not yet available. The present study was conducted to develop an animal model system by inoculating animals with a live Eimeria vaccine. This study was conducted on 3-day-old chickens (n = 20) pretreated with cyclophosphamide. The chickens were divided into 2 groups: the control group (n = 10) and the inoculated group that received the live Eimeria vaccine (n = 10). During the study period, we compared the clinical signs, changes in body weight, and number of oocysts shed in the feces of the control and inoculated group. This study showed that oocyst shedding was significantly higher in the chickens inoculated with live Eimeria oocysts than in the control chickens. Moreover, body weight gain was lesser in the animals in the inoculated group than in the control animals. Fecal oocyst shedding was observed in the inoculated animals. On the basis of these findings, we suggest that live Eimeria vaccination with cyclophosphamide pretreatment may be used to obtain an effective animal model for studying protozoan infections. This animal study model may eliminate the need for a tedious continuous animal inoculation process every 6 months because the live coccidiosis vaccine contains live oocysts.

• Korean Journal of Veterinary Service
• /
• v.21 no.4
• /
• pp.391-400
• /
• 1998

In order to survey the prevalence of intestinal parasites in dogs, 304 fecal samples were taken from dogs in Taejon city, The prevalence and identification of intestinal parasites were determined by the fecal examinations using sheather's floating technique and sedimentation methods and then Cryptosporidium oocysts were identified by kinyoun's modified acid fast stainning method. The results were obtained as follows ; 1. Parasite eggs and oocysts were detected in 105 samples (34.5%) from 304 cases of dog feces. 2. The 11 kinds of eggs and oocysts were isolated from the feces of dog. Those were Ancylostoma caninum (12.1%, 37 dogs), Trichuris vulpis (11.5%, 35 dogs), Toxocara canis (10.2%, 31 dogs), Isospora sp (7.2%, 22 dogs), Cryptosporidium sp (3.6%, 11 dogs), Toxascaris leonine (1.9%, 6 dogs), Strongyloides sp (1.9%, 6 dogs), Taenia sp (0.6%, 2 dogs), Diphylidium caninum (0.3%, 1 dog), Spirometra sp (0.3%, 1 dog) and Clonorchis sinensis (0.3%, 1 dog). 3. It was mixed infection such as single, double, triple and quadruple, 64.8%, 25.7%, 8.6% and 0.9%, respectively. 4. In indiviually-raised 4095, the infectious late of T canis (11.4%), A Caninum(13.2%), Cryptosporidium sp (6.1%), T leontna (2.6%) were higher than those of group raised dogs. But the infectious rate of T vulpis (12.1%) in group raised dogs was higher than that of individually-raised dogs. 5. Adults of Demodex and Sarcoptes which have been found in this survey are excluded in this report.