• Title/Summary/Keyword: modified acid fast stain

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Development of Cryptosporidium parvum in cell culture (세포배양에서 Cryptosporidium parvum의 발육)

  • Kim, Bo-sook;Joo, Hoo-don;Wee, Sung-hwan;Kim, Tae-jong
    • Korean Journal of Veterinary Research
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    • v.35 no.2
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    • pp.317-326
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    • 1995
  • The purpose of this study was to establish a method for in vitro culture of C parvum isolated in Korea by determination of suitable cell model to complete development of this parasite. The result obtained were summerized as follows: 1. To determine the most suitable cell line, six types of cell line were examined by microscopy. All cell lines were infected with C parvum and showed the highest infection score in HmLu cells. 2. The staining methods including DMSO-modified acid-fast(A-F) stain, hematoxylin-eosin(H & E) stain and immunofluorescence antibody(IFA) stain were applied to examine the infection of C parvum in cell culture. These staining methods were possible to examine the infection of C parvum in cell culture. The most sensitive one was IFA staining technique. 3. Developmental stages of C parvum in HmLu cell were observed. After the initial 8 hour incubation period, some trophozoites were observed. The meronts and gametes were appeared at 24-48 hour post inoculation(PI), and oocysts were observed firstly at 48-72 hour PI. 4. In H & E stain, the parasite appeared as basophilic within parasitophorous vacuole membrane(PVM) and lying in cytoplasm at near the nucleus of the host cells. It was able to distinguish the type I, type II meronts and gametes. 5. In DMSO-modified acid-fast stain, specific stained parasites were appeared firstly after 48 hour PI. The parasites were showed with different degrees of staining bright red color within PVM. 6. The endogenous stages of parasites in HmLu cell recovered at 48, 96, 120 and 144 hour after inoculation were reacted with rabbit immunized serum in immunofluorescence antibody and avidin-biotin complex peroxidase staining technique.

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Opportunistic Parasites among Immunosuppressed Children in Minia District, Egypt

  • Abdel-Hafeez, Ekhlas H.;Ahmad, Azza K.;Ali, Basma A.;Moslam, Fadia A.
    • Parasites, Hosts and Diseases
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    • v.50 no.1
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    • pp.57-62
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    • 2012
  • A total of 450 stool samples were collected from inpatient and outpatient clinics of Pediatric Department, Minia University Hospital, Minia District, Egypt. Two groups of patients were studied, including 200 immunosuppressed and 250 immunocompetent children. Stool samples were subjected to wet saline and iodine mounts. A concentration technique (formol-ether sedimentation method) was carried out for stool samples diagnosed negative by wet saline and iodine mounts. Samples were stained by 2 different methods; acid fast stain (modified Ziehl-Neelsen stain) and Giemsa stain. Total 188 cases (94%) were diagnosed positive for parasitic infections among immunosuppressed children, whereas 150 cases (60%) were positive in immunocompetent children ($P$<0.0001). The most common protozoan infection in immunosuppressed group was $Cryptosporidium$ $parvum$ (60.2%), followed by $Blastocystis$ $hominis$ (12.1%), $Isospora$ $belli$ (9.7%), and $Cyclospora$ $caytenensis$ (7.8%). On the other hand, $Entamoeba$ $histolytica$ (24.6%) and $Giardia$ $lamblia$ (17.6%) were more common than other protozoans in immunocompetent children.

An epidemiological survey of Cryptosporidium parvum infection in randomly selected inhabitants of Seoul and Chollanam-do (서울 및 전남 지역 주민의 작은와포자충 감염에 대한 역학조사)

  • 채종일;이상협
    • Parasites, Hosts and Diseases
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    • v.34 no.2
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    • pp.113-120
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    • 1996
  • An epidemiological survey was performed to know the status of Cwptespori,mum sp. infection among the people in Seoul and Chollanam-do in 1992. One village of Chollanam-do (Hwasun-gun) which showed the highest oocyst positive rate was re-surveyed in 1995 for human infection and for cattle also. The subjected areas consisted of 8 urban villages (: dongs) of Seoul and 4 urban (: dongs) and 7 rural (: myonsl villages of Chollanam-do. A total of 3,146 fecal samples was collected randomly, and smears were made from formalin-ether sediments. They were examined for Cwptosporinium oocysts by modified acid fast staining. The overall oocyst positive rate was 79% (248/3,146), but the rate was remarkably different between Seoul and Chollanam-do, 0.5% (4/853) and 10.6% (244/2,293), respectively The average size of oocysts was 4.8 ± 0.5 by 4.2 ± 0.5 ㎛, compatible with C. pcnlum. In Chollanam-do, rural villages showed significantly higher rate (14.0%) than urban villages (3.7%). Especially the people in Iyang-myon, Hwasun- gun, a typical rural village, revealed a very high rate of 40.0% (74/185). Adults aged 51-70 years revealed the highest positive rate among all age groups. At the re-survey of the same village of Hwasun-fun in 1995, 44 (35.2%) of 125 villagers and 14 (93.3%) of IS cattle examined were positive for C. pnnpum oocysts. The results suggest that C. pnnpum is highly prevalent in rural areas of Chollanam-do, and an important source or mode of infection seems to be contaminated water or contact with the feces of infected cattle.

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Prevalence of Cryptosporidium sp among calves and pigs in Jeonbuk province (전북지역의 소와 돼지에서 Cryptosporidium sp의 감염실태 조사)

  • 양홍지;김종승
    • Korean Journal of Veterinary Service
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    • v.24 no.2
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    • pp.133-138
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    • 2001
  • Cryposporidium species have a wide host range. These coccidian parasites are found in close association with epithelial cells of many species of animals including mm. The gastrointestinal tract is most commonly affected in young ruminants and this parasite is thought to be considerable importance in calf diarrhea complex. Major outbreaks of cryptosporidiosis have been reported in calves, lambs, pigs and others including avian species. Cryptosporidiosis is transmitted by oocysts of Cryptosporidium species. Because cryptosporidiosis is common infection among animals, early literature considered it a zoonosis. Human infections contracted from calves, cats, and horse feces. However, the resrvoir host is longer considered the major source of infection. Mild cases of disease have been reported in farm workers. Immunosuppressed, very young and very old persons should avoid contact with this parasite because it may cause severe diarrhea. In order to detect of Crytosporidium sp infection from feces of cattle and pigs at Chonbuk Iksan and Kunsan area, sedimentation and modified acid fast stain were applied. The positive rate of Cryptosporidium sp infection from 1, 176 of cattle and 267 of pigs were 0.5 % and 16.8%, respectively. According to area in Iksan and Kunsan, the positive rates were 0.4% and 0.9% from cattle, and 18.9% and 12.1% from pigs, respectively. In cattle, positive detection rate was 0.6% in milking cows but not in Korean cattle.

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Evaluation for detection of Cryptosporidium oocysts in diarrheal feces of calves (야외 송아지 설사변에서 작은와포자충 검출에 대한 평가)

  • Wi, Seong-Hwan;Ju, Hu-Don;Gang, Yeong-Bae
    • Parasites, Hosts and Diseases
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    • v.34 no.2
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    • pp.121-126
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    • 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.

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A Case of Nocardiosis (Nocardiosis 1예)

  • Kim, Jeong-Hee;Yoon, Ki-Heon;Yoo, Jee-Hong;Kang, Hong-Mo;Suh, Jin-Tae
    • Tuberculosis and Respiratory Diseases
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    • v.39 no.4
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    • pp.355-360
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    • 1992
  • Nocardiosis is an acute, subacute or chronic infection, which usually introduced through the respiratory tract resulting pneumonia, and may develop a disseminated infection, especially subcutaneous abscess and/or CNS infection. It is usually affects the immunocompromized host and may be fatal unless early diagnosis and adequate treatment are performed. There have been only several case reports of nocardial infection in Korea. Nocardiosis is so unfamiliar to many physicians that may be misdiagnosed as pneumonia, tuberculosis, or neoplasm. We have experienced a case of nocardiosis from a patient who had been treated as pneumonia and tuberculosis at first. The 57-year-old male patient had fever, chill, dyspnea and blood tinged purulent sputum for 20 days. Under the impression of bacterial pneumonia, broad spectrum antibiotics were administered for more than 3 weeks without clinical improvement. Although antituberculous drugs began to be administered after acid fast bacilli were found in bronchial aspirate by bronchoscopy, the nocardial infection was suspected due to no clinical response toward antituberculous therapy and the occurrence of multiple subcutaneous abscesses on scalp. The diagnosis was made by modified Ziehl-Neelson stain and culture of the sputum and pus. Nocardia asteroides was identified. After 25 days of trimethoprim-sulfamethoxazole treatment, the patient was much improved and discharged.

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