• 대한수의학회지
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• 제43권4호
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• pp.677-681
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• 2003

Currently, both the indirect fluorescent antibody test (IFAT) and enzyme linked immunosorbent assay (ELISA) have been used to detect Neospora caninum antibodies. Several factors such as the buffers, the conjugate, the pattern of fluorescence, and the cross reactivity with other apicornplexan protozoan, may result in poorly correlated data. The present study was undertaken to develop and evaluate the Neospora agglutination test (NAT) for the detection and quantification of IgG antibodies to N. caninum from various animal species. Compared to the ELISA method, the NAT with a cutoff value of 1:512 gave a high index of coincidence (kappa=0.807) and no cross reactivity to Toxoplasma gondii antiserum. Hence, this NAT method, which did not require a species-specific secondary antibody and expensive tools, would be easily available for the detection of antibodies to N. caninllm of various animal species.

• 대한수의학회지
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• 제49권4호
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• pp.291-295
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• 2009

A dead stray cat was necropsied for zoonotic feline disease monitoring. Grossly, there were no specific lesions. Major microscopic lesions included lymphocytic meningoencephalitis, malacia, and tissue cysts in the cerebral and cerebellar cortex. The size and shape of tissue cysts were identical to those of Apicomplexa including Toxoplasma (T.) gondii. Bradyzoites in the tissue cyst were strongly positive for T. gondii by immunohistochemistry. Electron microscopy revealed that bradyzoites within the tissue cyst were similar to the morphological features of T. gondii. Fresh tissue samples were examined by a polymerase chain reaction assay and resulted in a specific band of T. gondii only in the brain. Based on the results, this case was diagnosed as toxoplasmosis. This is the first case of toxoplasmic meningoencephalitis in a cat in Korea.

• Parasites, Hosts and Diseases
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• 제46권1호
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• pp.45-48
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• 2008

To evaluate the usefulness of the Korean Isolate-1 (KI-1) antigen for serodiagnosis of toxoplasmosis, antigen profiles of KI-1 tachyzoites were analyzed in comparison with RH tachyzoites by SOS-PAGE and immunoblotting. ELISA was performed on latex agglutination (LA)-positive and negative serum samples using KI-1 and RH antigens. Immunoblotting of the KI-1 antigen showed multiple antigen bands with molecular sizes of 22-105 kDa. Among them, 1 and 6 common bands were noted against a KI-1-infected and a RH-infected human serum, respectively, which represented differences in antigenic profiles between KI-1 and RH tachyzoites. However, all 9 LA-positive human sera were found positive by ELISA, and all 12 LA-negative sera were negative by ELISA; the correlation between the ELISA titers and LA titers was high (r = 0.749). Our results suggest that tachyzoites of KI-1 may be useful for serodiagnosis of human toxoplasmosis.

• 대한수의학회지
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• 제15권2호
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• pp.309-314
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• 1975

In survey for internal parasites of 395 heads of swine by fecal examination at ict, incidences of each parasite were obtained as follows: Giardia lamblia 1.0% Entamoehaspp. 55.4 Eimeria& Isospora spp. 22.5 Balantidium coli 66.6 Metastrongylus elongatus 17.6 Ascaris suum 25.6 Oesophagostomun dentatum 29.1 Hyostrongylus ryubidus 14.6 Trichuris suis 4.2 Strongyloides ransomi 7.2 Mecistocirrus digitatus 1.0 Check-list for the internal parasites of swine made by all the materials repor years from 1920 to 1975 in Korea is as follows: No. Parasites Habitat References 1. Ascaris lumbricoides small intestine Kawamura(1923) 2. Oesophagostomum dentatum large intestine Kawamura(1923) 3. Echinococcus veterinorum lung & liver Kawamura(1923) 4. Cysticercus cellulosae muscle Yunoba(1923) 5. Sarcooystis sp. muscle Arahayase(1927) 6. Entamoeba polecki intestine Kuwabara(1931) 7. Balantidium coli large intestine Huruyama(1931) 8. Metastrongylus elongatus lung Lee(1956) 9. Gongylonema pulckrum oesophagus Isshiki(1960) 10. Ascarops strongylina stomach Isshiki(1960) 11. Cysticercus tenuicollis peritoneum Isshiki(1960) 12. Cysticercus bovis? diaphragm Isshiki(1960) 13. Toxoplasma gondii interna organs Mun(1960) 14. Trichuris suis large intestine Lee et al.(1963) 15. Stephamirus dentatus feces Lee et al.(1963) 16. Spirometra mansonides fat layer of muscle Jang(1964) 17. Hyostrongylus rubidus stomach Kim et al.(1969) 18. Strongyloides ransomi feces Kim et al.(1969) 19. Eimeria perminuta feces Jang(1972) 20. E. debrieki feces Jang(1972) 21. E. polita feces Jang(1972) 22. E. scabra feces Jang(1972) 23. E. scrofae feces Jang(1972) 24. Isospora suis feces Jang(1972) 25. Entamoeba coli feces Jang(1975) 26. Mecistocirrus digitatus feces Jang(1975) 27. Giardia lamblia feces Jang(1975).

• 한국동물위생학회지
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• 제34권4호
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• pp.303-311
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• 2011

Prevalence of infectious agents, including Brucella abortus (BA), Mycobacterium bovis (MB), bovine leukemia virus (BLV), M. avium subsp. paratuberculosis (MP), Neospora caninum (NC) and Toxoplasma gondii (TG), was investigated in all the cattle raised in Ulleung island during 2007~2010. For BA, the prevalences in head and farm were 8.1% (44/545) and 5.5% (4/73) in 2007, all negative in 2008~2009, and 0.5% (4/774) and 1.7% (1/58) in 2010, respectively. For MB, no sample was positive by PPD or ELISA in 2007~2010. For BLV and MP, no sample was positive by ELISA in 2007~2009. For NC, seroprevalences in head and farm were 0.2% (1/545) and 1.4% (1/73), respectively, in 2007 and all negative in 2008~2009. For TG, seroprevalences in head and farm were 17.6% (97/552) and 54.8% (34/62) by ELISA in 2009. By regions, the seroprevalences of TG in Ulleung-eup, Seo-myeon and Buk-myeon were 26.0%, 9.8% and 16.7%, respectively, which had significant differences (P<0.0001). Tiger cattle were more resistant to TG infection than Hanwoo. The seroprevalence of TG in summer was higher than in autumn. The seroprevalence of TG in cows was higher than in oxen. The seroprevalence of TG in cattle was increased with age. In conclusion, this study indicates that the prevalences of six infectious diseases, except for TG which are widely spread, are relatively low in cattle reared in Ulleung island.

• 대한의생명과학회지
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• 제9권2호
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• pp.85-91
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• 2003

Gentiana scabra var. buergeri (G. scabra) is a herb known to have therapeutic effect in infection diseases. We studied cellular activation and antitoxoplasmosis in macrophages after G. scabra stimulation. Macrophage activation was detected by nitrite production. Macrophages were treated with G. scabra extracted with water or methanol. Maximal nitrite production was detected in macrophages after stimulation of G. scabra extract 0.1 mg/ml. Maximal nitrite concentration was 23.22 0.003 uM/L in macrophages after water extract of G. scabra and was 24.07 1.41 uM/L after methanol extract of G. scabra. Effect of G. scabra in the phagocytic capacity of macrophages was monitored by using PI (percentage of macrophage infected by T gondii) method. The minimum PI (42.5 2.31) was detected in macrophages treated by water extract of G. scabra 0.1 mg/ml before infection of T gondii. We also examined toxoplasmastatic capacity of macrophage using FI (fold increase) method. The minimum FI (4.46 1.16) was shown in macrophages after water extract of G. scabra 0.1 mg/ml pretreatment before infection. Under electron microscope, proliferation of T gondii was inhibited by extract of G. scabra treatment in macrophages and the mitochondrion and lysosomal vacuoles within cells were increased. Taken together, G. scabra extract activates macrophages and induces toxopalsmastatic activity after T gondii infection. It is suggested that G. scabra may be used as a therapeutic drug against toxoplasmosis.

• Parasites, Hosts and Diseases
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• 제39권3호
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• pp.241-246
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• 2001

The antigenic domain of the major surface protein (Nc-p43) of Neospora caninum was examined by polymerase chain reaction of its gene fragments and recombinant expression as GST fusion proteins. The fragments of Nc-p43 were as follow: a total open reading frame (OFR), T: OFR without signal sequence and C-terminal hydrophobic sequence, S: N-terminal 2/3 parts of S, A: C-terminal 2/3 parts, P; N-terminal 1/3 part, X: middle 1/3 part Y; and C-terminal 1/3 part, Z, respectively. The DNA fragments were cloned into pGEX-47 vector. Recombinant plasmids transformed into Escherichia coli of BL21 pLysS (DE3) strain were induced to express GST or GST fused fragments of Nc-p43 such as 69 kDa protein for T,66 kDa for S, 52 kDa for A,53 kDa for P, and 40 kDa proteins for X, Y, and Z, respectively in SDS-PAGE. The Nc-p43 fragments of T, S, and P reacted with a bovine serum of neosporosis while those of A, X, Y, and Z together with GST did not in the western blot. These findings suggest that the antigenic domain of Nc-p43 of N. caninum may be localized in the C-terminal 2/3 parts. Together with Al9 clone in SAGI of Toxoplasma gondii (Nam et at., 1996), the P fragment of Nc-p43 could be used as efficient antigens to diagnose and differentiate those infections with both species .

• Parasites, Hosts and Diseases
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• 제57권2호
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• pp.83-92
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• 2019

Based on the reactive oxygen species (ROS) regulatory properties of diphenyleneiodonium (DPI), we investigated the effects of DPI on host-infected T. gondii proliferation and determined specific concentration that inhibit the intracellular parasite growth but without severe toxic effect on human retinal pigment epithelial (ARPE-19) cells. As a result, it is observed that host superoxide, mitochondria superoxide and $H_2O_2$ levels can be increased by DPI, significantly, followed by suppression of T. gondii infection and proliferation. The involvement of ROS in anti-parasitic effect of DPI was confirmed by finding that DPI effect on T. gondii can be reversed by ROS scavengers, N-acetyl-L-cysteine and ascorbic acid. These results suggest that, in ARPE-19 cell, DPI can enhance host ROS generation to prevent T. gondii growth. Our study showed DPI is capable of suppressing T. gondii growth in host cells while minimizing the un-favorite side-effect to host cell. These results imply that DPI as a promising candidate material for novel drug development that can ameliorate toxoplasmosis based on ROS regulation.

• Parasites, Hosts and Diseases
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• 제34권3호
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• pp.197-206
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• 1996

율무 수침엑스(Co-Ex)가 대식세포내 톡소포자충(Toxoplosmo gondii RH strain)의 번식에 억제 작용이 있다는 보고(Soh et al.., 1994)에 이어 본 연구는 Co-Ex내의 유효성분(effective component)을 추구하기 위하여 이미 탈지조작과 단백질 변성과정을 거친 Co-Ex를 Tris-buffer solution에 녹인 후 원심과정으로 그 상청(WC1) 그리고 WC1을 ammonium sulfate 처리 후 원심한 상청(WC2)과 침사(WC3)를 분리하였고 WC1은 다시 column chromatography에 의하여 WC4 WC5 WC6로 분획하여 시료로 사용하였으며. 대식세포 활성에 기여하는 것으로 이미 알려진 $IFN-{\gamma}$, LPS 등을 참고자료로 하였다. 각 시험자료들의 대식세포 활성화로 인한 반응질소 중간산물(RN1) 생산량. 대식세포의 톡소포자충 감염상 대식세포내 톡소포자충 번식상 등을 비교 조사하였다 10% FCS 첨가 DMEM 배지에 배양하였고 이에 각종 시료(BRM)를 실험목적에 따라 배지에서 24시간 작용시킨 뒤 RNI를 조사하였고 이어 톡소포자충 $1\times10^6$을 30분 감염시킨 뒤 새 배지에 옮겨 48시간 배양한 뒤 Pl, Fl 등을 조사하였다. NO 생산은 각례 대조(시료 비투여)에 비하여 높았고 $IFN-{\gamma}$ 첨가는 생산량(${\mu}M/l$)을 더욱 증가시켰다(예 Control 7.5 Control + $IFN-{\gamma}$ 14.2 WC2 26.2 WC2 + $IFN-{\gamma}$ 57.8). 포자충감염율(%)은 대조에 비하여 낮았고 $IFN-{\gamma}$ 첨가시에는 더욱 낮아졌는데 $IFN-{\gamma}$ 첨가예에서의 감염율은 대조 38.0인데 비하여 WC2 19.2. WC1 + WC2 + WC3 7.2로 나타났다. 세포당감염율도 같은 경향으로 INF-$\gamma$ 첨가시험에서 대조 6.0(감염 대식세포 100개 중의 톡소포자충 평균수) WC2 1.7, WCI + WC2 + WC3 1.4 등으로 나타 났다. 결론적으로 율무엑스 수침엑스 중 WC2, WC1 등은 대식세포를 활성화하나 $IFN-{\gamma}$ 첨가는 더욱 그 활성화를 높이며 그 중 WC2가 활성화에 주역을 하는 것으로 사료되는 바이다.

• Parasites, Hosts and Diseases
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• 제35권4호
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• pp.251-258
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• 1997

톡소포자충. RH주 tachyzoite의 항원성 난백질에 대하여 알아보고자 마우스 복강 내 또는 in uipo에서 Hep-2 cell에 계대한 톡소포자충과 감염마우스의 복팡앤으로 SDS-PAGE/immunoblot을 시행하였다 또 lmnoanity chromatoaphy를 이용하여 톡소포자충 항원을 징제한 후 SDS PAGE/immunoblot을 시행하여 항원을 분리하였으며. 톡소포자충 조항원 및 정제항원으로 면역시킨 마우스 혈청을 이용하여 IgG-ELISA를 시행하였다. 톡소포자충 용해물을 면역시킨 노끼의 항혈 청으로 immunoblot을 시행하였을 때 마우스 복강 내로 계대한 톡소포자충에서 76 kDa. 70 kDa. 64 kDa. 53 kDa 46 kDa. 44 kDa. 35 kDa. 25 kDa. 18 kDa 및 13 kDa의 항원대가 관찰되었으며. in vitro에서 Hep-2 cell에 배양한 톡소포자충은 70 kDa. 64 kDa. 53 kDa 35 kDa. 25 kDa 및 13-10 kDa의 항원대가 관찰되어 마우스 복강 내에 계대한 톡소포자충에서 더 많은 항원대가 관찰되었다. 마우스 복강 계대한 tachyzoite 용해물을 immunoaffinity글 시행하여 부분 정제한 후 떤역시킨 초기의 항혈청으로 immunoblot을 하였을때 E-1에서는 97 kDa. 63 kDa. 53 kDa 및 35 kDa이 E-2에서는 53 kDa 및 35 kDa이 나타났다. 한편 감염 마우스 복강 액에서는 76 KDa 53 KDa. 35 kDa 및 29-28 kDa의 항원대가 관찰되었으며. 정상 마우스 복강 액에서도 약하게 76 kDa .35 kDa 및 29-28 kDa의 반응대가 관찰되었다. 감염 마우스 복강앤을 IgG-Sepharose column을 통과시킨 후 시행한 immunoblot에서 E-1은 84 kDa. 76 kDa. 5,B kDa 및 29 kDa이 . E-2에서 53 kDa 및 45 kDa의 항원대가 관찰되었다. 이 실험에서 immunoawnity chromatography를 이용하여 톡소포자충 용해물 및 분비물에서 76 kDa. 63 kDa. 53 kDa. 35 kDa 및 29 kDa의 항원을 불리하였다. 톡소포자충의 조항원과 정제항원 (감염 마우스 복강액. E-1)으로 IgG-ELISA를 시행한 결과. 조항원의 경우 2차 면역 후 1주 후부터 IgG 항체가 증가하였고 정제 항윈은 2차 면역 후 3주 후부터 IgG항체가 증가하였다 (p<0.05).