한우 및 이상산 송아지의 Neospora caninum의 감염에 대한 면역조직화학 적 및 혈청학적 관찰

손성봉 ( Seong Bong Son ),정원일 ( Won Il Jeong ),정규식 ( Kyu Shik Jeong ),이차수 ( Cha Soo Lee ) 한국동물위생학회 2004 한국동물위생학회지 (KOJVS) Vol.27 No.1

The present study was carried out to investigate the prevalence of Neospora caninum infection in Korean indigenous cattle and calves of abnormal deliveries and focus on correlation between malformation and N caninum infection. To determine the prevalence of antibodies to N caninum, sera of 473 Korean indigenous cattle from slaughter house were tested for N caninum antibodies using indirect fluorescence assay. Of the 473 cattle sera, 9.5% (45/473) showed positive against N caninurn Regional seropositive rates of the samples were 16.7% (5/85), 11.0% (11/100), 8.8% (21/240) and 5.9% (5/85) at Kyonggi, Gyeongbuk, Daegu and Kyongnam province, respectively. In female, seropositive rates were 17.5% (25/143) and 6.1% (20/330) in male. During the period from march 2000 to August 2001, 55 abnormal deliveries of Korean indigenous cattle including abortion, stillbirth and congenital malformation were examined by histopathological, immunohistochemical and serological methods for evidence of N caninum infection. Of the 55 abnormal deliveries, only 5 calves showed positive reaction against N caninum in serological test. In microscopical observation, gliosis and nonsuppurative myositis were observed. However, Neospora-like organisms were not detected by either periodic acid-schiff (PAS) reaction or immunohistochemical technique. Taken together all these data, this study indicate that N caninum infection was widespread in breeding farms of Korean native cattle, caninum infection was not recognized, but correlation between malformation and N.

정읍지역에서 사육중인 한우에서 Neospora caninum 항체 양성율 조사

정재명 ( Jae Myong Jeong ),권미순 ( Mi Soon Kweon ),윤여백 ( Yeo Baik Yoon ),한규삼 ( Kyu Sam Han ) 한국동물위생학회 2005 한국동물위생학회지 (KOJVS) Vol.28 No.2

This survey was carried out to investigate the seroprevalence of antibodies to Neospora caninum in Korean indigenous cattle that was representative livestock raised in Jeongeup province Jeonbuk Korea. A total of 1,162 sera were tested for N caninum antibodies using ELISA(Herdcheck anti-Neospora, IDEXX Laboratories Inc., Westbrook, Maine USA). 15(1.3%) sera were positive by ELISA. A total of 290 farms were tested, 11(3.8%) farms were positive. Among the seroprevalence of cattle according to the areas, 8 of 44 counties were infected N caninum. Sero-positive 11 farms of N caninum antibodies using ELISA test and 62 of 301 sera (20.6%) were infected to N caninum. We found that cattle be raised in Jeongeup province was slightly infected to N caninum. It seems to be infected of N caninum an early stage, but the positive rates was variable that it was 6%~62%. It was supposed to the highest positive rate of 3 farms had some factors to be infected. The factors were supposed to keep dogs yearly, located the base of hills, and almost feed with imported dried grass. Farmers suppose to the cause of abortion was not the neosporosis but also broke out an accident or was infected Brucella spp. The results of Brucella Rose Bengal Test(RBT) were all negative.

공주, 연기지역 젖소의 Neospora caninum 항체 양성율 및 갓 태어난 송아지의 감염양상 조사

손정훈,박배근,서상희,손화영,조성환,류시윤 한국임상수의학회 2011 한국임상수의학회지 Vol.28 No.1

To examine the seroprevalence of Neospora caninum (N. caninum) infections in cattle raised at farms where an abortion had previously occurred in Gongju city and Yeongi-gun of Chungnam Province, 280 head of cattle from six farms were examined by ELISA. Thirty-two pregnant cattle from four farms were examined to determine the abortion rate in N. caninum-infected pregnant cattle and the infection pattern in newborn calves. The six farms where the abortions had previously occurred demonstrated positive reactions to N. caninum. Although the mean seropositive rate of the six farms was 17.5%, the range of seropisitivity was 3.4-57.1% due to the difference in seroprevalence between farms. The general pattern of seropositivity in cows was higher than in heifers. In addition, the abortion rate of N. caninumpositive in 18 out of 32 pregnant dams raised in four farms was 16.7%. The seroprevalence of newborn calves from N. caninum-positive pregnant animals differed according to the farm but was 86.7% overall. In contrast, 14 newborn calves from the N. caninum-negative pregnant cattle showed negative responses. This suggests that N. caninum is endemic and occurs through vertical transmission from chronically-infected dams. Therefore, aborted dams should be discarded.

Neospora caninum에 의한 젖소의 반복유산

김재훈,황의경,손현주,진영화,윤순식,김대용,Kim, Jae-hoon,Hwang, Eui-kyung,Sohn, Hyun-joo,Jean, Young-hwa,Yoon, Soon-seek,Kim, Dae-yong 대한수의학회 1998 大韓獸醫學會誌 Vol.38 No.4

A case of repeated bovine abortion caused by Neospora caninum is described. The 3-year-old Holstein cow, which was seropositive to N caninum antibody by indirect immunofluoresent assay, aborted consecutively within a 10-month period. Two fetuses of 6- or 5-month of gestation were aborted in June 1996 and March 1997, respectively. Histologically, multifocal necrotizing encephalitis, nonsuppurative periportal hepatitis, myocarditis, and myositis were observed in both fetuses. Tachyzoites or tissue cysts detected in the brain, heart and liver were positive to N caninum but negative to Toxoplasma gondii by immunohistochemical method. The results of our study demonstrate that repeated abortion due to N caninum actually occurs in Korea. This is believed to the first report of repeated abortion associated with N caninum in Korea.

Neospora caninum에 자연 감염된 임신우로부터 유산된 태아의 병리조직학적 관찰

손정훈(Jeong-Hoon Son),조성환(Sung-Whan Cho) 한국생명과학회 2010 생명과학회지 Vol.20 No.10

충남 공주시와 연기군의 목장 4곳에서 Neospora에 자연 감염된 임신우로부터 유산된 태아를 병리조직학적, 전자현미경적으로 관찰하였다. 유산된 태아는 전신 부종과 함께 부검시 복강과 흉강은 혈액성ㆍ장액성 액체로 충만되어 있었다. 광학현미경적으로 심장과 간에서는 충ㆍ출혈 및 심근세포와 간세포의 괴사를 동반한 대식 세포, 임파구, 단핵구 세포로 구성된 많은 수의 염증세포 침윤이 관찰되었고, 간에서는 간세포의 세포질과 간질 조직에서 tachyzoite 집락이 관찰되었다. 뇌에서는 신경세포내와 주위에서 크기가 다양한 많은 수의 조직낭포가 관찰되었고, 조직낭포는 구형으로 두꺼운 막으로 싸여 있었으며 내부에는 많은 bradyzoite를 포함하고 있었다. 또한, 뇌조직에서는 충ㆍ출혈 및 국소성 괴사와 함께 광범위한 신경교증이 관찰되었고 국소성 괴사 부위 주위와 혈관주위에 소교세포로 구성된 많은 수의 염증세포 침윤이 관찰되었다. 전자현미경적으로 조직낭포의 막 두께는 대략 1 ㎛ 로 모양은 불규칙하였고, 내부에는 길이 2-5 ㎛, 폭 1-2 ㎛인 100여 개의 bradyzoite가 관찰되었다. Bradyzoite의 핵은 posterior tip으로부터 1-1.5 ㎛ 전방에 존재하였고, amylopectin 과립, 전자밀도가 높고 작은 구형인 과립과 전자밀도가 상대적으로 낮으나 큰 구형의 과립, 전자밀도가 균질한 rhoptry 및 zoite pellicle에 대해 수직방향으로 달리는 microneme이 핵과 posterior tip 사이의 세포질 내에서 관찰되었다. 이상의 본 연구는 ELISA검사를 통해 Neospora 항체에 양성반응을 나타내는 임신우로부터 유산된 태아에서 발견된 조직낭포를 전자현미경적으로 관찰하여 N. caninum임을 확인하였다. This study examined the histopathologic and electron microscopic findings of aborted fetuses from pregnant dairy cows naturally infected with Neospora caninum (N. caninum) at four farms in Gongju city and Yeonki gun of Choongnam province. Systemic subcutaneous edema was observed in the aborted fetuses. The necropsy revealed considerable serosanguinous fluid in the body cavity of the aborted fetuses. Light microscopy showed the infiltration of many inflammatory cells consisting of macrophages, lymphocytes and mononuclear cells, accompanied by congestion, hemorrhage and necrosis of myocardiac cells and hepatocytes in the liver and heart of the aborted fetuses. In the liver, clusters of tachyzoites were formed in the cytoplasm of hepatocytes and the interstitial tissue. In the brain, many tissue cysts of various sizes were observed in the nerve cells and their adjacent areas. Tissue cysts had a round shape and contained a large amount of bradyzoite. In addition, there was diffuse gliosis accompanied by congestion and hemorrhage and focal necrosis in the brain. Infiltration of microglial cells were observed at the periphery of the focal necrosis and perivascular area in the brain. Electron microscopy showed that the tissue cyst wall had a thickness of approximately 1 ㎛ with an irregular shape. On the interior side, more than 100 bradyzoites with lengths of 2-5 ㎛ and widths of 1-2 ㎛ were observed. The nucleus of in the bradyzoites was located approximately 1-1.5 ㎛ anterior to the posterior tip of the zoite. In the cytoplasm between the nucleus and the posterior tip, there were many amylopectin granules, electron-dense small-sized and electron-thin large-sized round granules, homogeneously electron-dense rhoptries and micronemes oriented perpendicularly to the zoite pellicle. To summarize, tissue cysts were identified on electron microscopy from the aborted fetus from N. caninum seropositive pregnant cow by the ELISA. This led to the confirmed presence of N. caninum.

Fecal and Molecular Survey of Neospora caninum in Farm and Household Dogs in Mashhad Area,Khorasan Province, Iran

Gholamreza Razmi 대한기생충학열대의학회 2009 The Korean Journal of Parasitology Vol.47 No.4

Neospora caninum is an important cause of abortion in dairy cattle worldwide. Dog is the definitive host for N. caninum and can infect dairy cattle. The aim of this study is to determine the prevalence of Neospora oocysts in feces of dogs from dairy farms. A total of 174 fecal samples was collected from 89 farm dogs and 85 household dogs during 2006 and 2008. Fecal samples of dogs were microscopically examined for detecting Hammondia Neospora-like oocysts (HNLO) by Mini Parasep??SF fecal parasite concentrator. HNLO were microscopically detected in 4 fecal samples (2.2%). The fecal samples with HNLO were examined by N. caninum-specific PCR. Two of the samples were positive for N. caninum. The 2 positive fecal samples were selected for inoculation to calves. Two inoculated calves were seronegative by ELISA for 4 months post-infection. This is the first report of finding N. caninum DNA in feces of farm dogs in Mashhad area, Iran.

국내 소에서 Neospora caninum의 분리

김재훈,손현주,황의경,황우석,허권,진영화,이병천,이재진,강영배,야마네 이츠로,김대용,Kim, Jae-hoon,Sohn, Hyun-joo,Hwang, Eui-kyung,Hwang, Woo-suk,Hur, Kwon,Jean, Young-hwa,Lee, Byung-chun,Rhee, Jae-chin,Kang, Yung-bai,Yamane, Itsuro,Kim, 대한수의학회 1998 大韓獸醫學會誌 Vol.38 No.1

The Neospora sp. was isolated from the brain of 1 calf via continuous in vitro cultivation in Vero cell. Neospora tachyzoites were observed 45 days after inoculation of the homogenized brain suspension into the Vero cell. The isolated parasite (named tentatively as NCKB-1) was morphologically and ultrastructurally similar to the previously reported Neospora sp isolated in cattle (BPA-1, JPA-1). A comparison of the antigenic reactivity of cultivated tachyzoites with polyclonal antisera to Neospora caninum and Toxoplasma gondii confirmed that this protozoal isolate was similar to N caninum. This is the first report of successful isolation of Neospora sp from cattle in Korea.

소에서 Neospora caninum에 대한 항체가 조사

허인 ( In Heo ),김영진 ( Young Jin Kim ),김희 ( Hui Kim ),허진희 ( Jin Hoi Heo ),박일규 ( Il Gyu Park ),강승원 ( Seung Won Kang ),정우석 ( Woo Seog Jeong ) 한국동물위생학회 2001 한국동물위생학회지 (KOJVS) Vol.24 No.1

This study was carried out to investigate the prevalence of Neospora caninum infection in dairy cow and Korean native cattle(KNC), raised in several Chungnam province. To determine the prevalence of antibodies to N caninum, a total of five hundred fifty six sera were analyzed by indirected fluorescent antibody (IFA) test. Five hundred thirty three sera were collected from fifteen dairy herds and twenty three sera were taken from fourteen KNC herds from December 1999 to November 2000. Seropositive ratio of the dairy cattle sera were individually or herdly tested and showed 64.2% and 93.3%, respectively. It was recorded with 78.6% and 47.8% in KNC. The seropositive ratio of dairy cattle was depended on the size of ranch. It was 92.2, 60.7 and 57.9% at the size of less than thirty, thirty to seventy and more than seventy one cattle, respectively. However, it was different from the province of Chungnam. The seropositive ratio to N caninum of dairy cattle were 79.5, 53.1, 61.4 and 31.1% at Gongju, Yeongi, Geumsan and Cheongwon, respectively. It showed difference at the growth stage and sex of cattle. The seropositive ratios of N caninum of calf, heifer, premiparous, multiparous (2nd-5th), multiparous(6>th) and bulls confirmed to 25.0, 50.3, 70.3, 71.2, 50.0 and 50.0%, respectively. It was related with brucellosis in cattle. The infected ones with brucellosis were 75.7% of seropositive ratios to N caninum. The results of this study indicated that N caninum infection was widespread in Chungnam province and confirmed existing with brucellosis in cattle.

파라핀 블록 PCR을 이용한 소 네오스포라 감염증의 진단법 확립

이중근,김재훈,김진현,이병천,황우석,윤희정,남호우,진영화,김대용,Lee, Jung-keun,Kim, Jae-Hoon,Kim, Jin-Hyun,Lee, Byung-chun,Hwang, Woo-suk,Youn, Hee-jeong,Nam, Hoo-woo,Jean, Young-wha,Kim, Dae-Yong 대한수의학회 2001 大韓獸醫學會誌 Vol.41 No.3

Neospora caninum infections have been associated with neonatal paresis as well as abortion around the world. Bovine abortion induced by N caninum was first reported in 1997 in Korea. Diagnosis of N caninum infection is usually based on histopathology and immunohistochemical detection of organism. However, often the tissues having lesion suggestive of N caninum infection were negative on immunohistochemistry. Here, we describe establishment of PCR-based diagnostic strategy for N caninum infection using DNA extracted from paraffin blocks containing the lesion. PCR was able to amplify N caninum-specific bands from the paraffin blocks containing at least moderate degree of inflammation. Compared to paraffin-blocks, DNA extracted from fresh tissues were less sensitive than that of paraffin blocks. This PCR-based method can be practically applicable for rapid diagnosis of bovine N caninum infection with high specificity and sensitivity. Based on this method, 17% of bovine abortion surveyed during a designated period was associated with N caninum infection.

Relationship Between IFNγ Production, Antibody and Hormone Levels in Naturally Neospora caninum-infected Pregnant Dairy Cows

손정훈,박배근,손화영,정주영,박상준,김태환,조성환,류시윤 한국임상수의학회 2010 한국임상수의학회지 Vol.27 No.6

Neosporosis is a widespread parasitic disease caused by Neospora caninum, an intracellular protozoan parasite. It causes economic losses due to reproductive failure. The potential relationship between pregnancy outcomes and levels of IFNγ, hormones, and antibodies in naturally N. caninum-infected cows was examined in the blood samples collected every 2 or 4 weeks in 26 pregnant cows from 4 different farms. The mean S/P value of seropositive nonaborting animals (n = 14) reached peak levels 15 weeks prior to parturition, and declined thereafter to parturition. The S/P value 13 weeks prior to abortion in seropositive aborting cows (n=3) remained at high levels, and abortions occurred at 20 (142 days), 26 (185 days), and 28 weeks (199 days) after artificial insemination. IFNγ levels in the seropositive non-aborting group varied by individuals and gestational periods; IFNγ levels stayed at elevated levels or increased abruptly close to abortion in seropositive aborting cows. IFNγ level patterns in the seronegative group (n = 9) were similar to the seropositive non-aborting group, although IFNγ amounts were lower than the seropositive group. The mean progesterone levels in the seropositive non-aborting and seronegative groups decreased markedly 7 weeks prior to parturition. The mean progesterone levels 5 and 7 weeks prior to abortion were lower than the other groups 5and 7 weeks prior to parturition. The mean 17β-estradiol levels in the seropositive aborting cows increased close to abortion; the produced amounts were lower than those of seropositive non-aborting and seronegative groups close to parturition. These results suggested that lower levels of progesterone and β-estradiol in Neospora-infected cows may lead to increases in IFNγ production and in turn may result in abortion.