시험관내에서 인공배양한 제 3기 자충 및 성충을 이용한 구충효능 선발시험

지차호,박승준,Jee, Cha-ho,Park, Seung-jun 대한수의학회 1998 大韓獸醫學會誌 Vol.38 No.3

The in vitro screening tests against the in vitro cultivated $L_3$ of Ascaris suum (in vitro $L_3$), which were cultivated from the embryonated egg to third-stage larva on 7 days in culture(DIC) and the in vivo rat's lung-derived $L_3$ of Ascaris suum (in vivo $L_3$), which were recovered from the lungs of rat on 7 days after infection, carried out in order to compare the anthelmintic efficacy of in vitro $L_3$ and that of in vivo $L_3$ in RPMI medium 1640 with 5% bovine calf serum. And also a screening test of efficacy against adult worms of Trichuris suis performed. The efficacies of screening tests were as follows : 1. The screening efficacies of abamectin and ivermectin against the in vitro $L_3$ were all 100% at the 10ppm concentration in RPMI medium 1640 on 5 DIC. 2. The screening efficacies of abamectin and ivermectin against the in vivo $L_3$ were all 100% at the 20ppm on 5 DIC or at 40ppm on 3 DIC. 3. The screening efficacies of abamectin and ivermectin against the adult worms of Trichuris suis were all 100% at 20ppm on 4 DIC. And therefore, the in vitro cultivated $L_3$ of Ascaris suum were used in the screening test as well as the in vivo rat's lung-derived $L_3$ of Ascaris suum. And also the adult worms such as Trichuris suis and filaroids which is small size and difficult to cultivate to vitro, were used in the screening test in vitro.

제1위 섬모충(rumen ciliates)을 이용한 동물성 단백질(치어용 사료) 개발

지차호,현공율,Jee, Cha-ho,Hyun, Gong-yool 대한수의학회 1995 大韓獸醫學會誌 Vol.35 No.2

This study was carried out to develop the animal protein(feed for fry) that was isolated, purified and lyophilized the rumen ciliates from the healthy rumen contents which have $10^5-10^6/g$ ciliates and were discarded in abattoirs. The rumen ciliates are non-pathogenic, anaerobic and the weight of this protozoa is 2% of rumen content. The rumen protozoan and bacterial proteins both have a biological value for rats of 80-81, which is higher than the 72 of brewer's yeasts. Furthermore, the true digestibility and net protein utility of the protozoan protein are 91 and 73, much higher than those of bacterial(74 and 60) or yeast(84 and 60) proteins. The amino acids of rumen protozoa is nutritionally superior than the others. The size of rumen ciliates is $30-200{\times}20-110{\mu}m$ and so we had isolated and purified the rumen ciliates from the rumen contents by the physical methods. The purified rumen protozoa was lyophilized with freezing dryer. The results of this experiment were as follows : 1. Population dynamics of protozoan ciliates in slaughtered rumens; % of samples which small ciliates were predominated was 82.5%(52/63) and that of large ciliates was 17.5%(11/63). 1) predominant species of small ciliates were Entodinium ovinum and E nanellum. 2) predominant species of large ciliates were Epidinium ecaudatum and Diploplastron affine. 2. The lyophilized rumen ciliates which were isolated and purified from 1 kg of rumen content at the pH 6.2-6.8 was about 7.0 gram. 3. The nutrient analysis of lyophilized rqmen ciliates(LRC) was as follows: 1) Proximate analysis of the LRC and the composition of fry feed; moisture 8.05%(below 10.0), protein 35.37%(45), fat 5.39%(4.5), fiber 1.23%(below 2.5), ash 2.25%(below 15.0), Ca 0.26%(below 2.0), P 0.14%(below 1.1), energy 4,608.11(fish meal 5000 cal/g) 2) Amino acids (% in crude protein) of the LRC and the rotifer(Brachionus plicatilis); Arg 5.19%(4.50), His 2.50%(1.55), Ile 5.29%(3.45), Leu 8.11%(5.85), Lys 10.34%(6.15), Met 2.25% (0.85), Phe 5.66%(3.80), Thr 5.14% (3.45), Val 4.18%(3.90), Ala 4.13%(3.35), Asp 13.26%(8.25), Glu 16.62%(9.20), Gly 4.23%(3.10), Pro 3.25%(5.05), Ser 4.85%(3.85), Tyr 5.04%(3.05) 3) Fatty acids(% in fat) of the LRC and the rotifer(biological feed ; Brachionus plicatilis); myristic acid(C14:0) 3.27%(0.3), myristoleic acid(C14:1) 0.83%(-), palmitic acid(C16:0) 39.11% (23.5), palmitoleic acid(C16:1) 2.81%(2.0), stearic acid(C18:0) 9.36%(5.6), oleic acid(C18:1) 25.54%(3.5), linoleic acid(C18:2) 15.05%(32.9), linolenic acid(C18:3) 1.74%(9.8). Judging from the above investigated results, the analytical data of proximate analysis, amino acids, fatty acids of the purified and lyophilized rumen protozoa are reasonable for the feed of freshwater fishes(fry and fingerling). But it was disappointed of our expectation that the crude protein of lyophilized rumen ciliates contains low percentage, it was thought that because of the small ciliates(starch digester) in beef cattle rumens which were administered the concentrated feed, is much difficult to isolate and purify than the large ciliates(fiber digester).

시험관내에서 돼지회충(Ascaris suum) 함자충란(L₂)의 인공배양

지차호,박승준,Jee, Cha-ho,Park, Seung-jun 대한수의학회 1998 大韓獸醫學會誌 Vol.38 No.1

The cultivation for development of Ascaris suum from the second-stage larvae($L_2$) embryonated egg and the third-stage of rat-derived larvae($L_3$) recovered from lung of rats were performed to use the screening test of anthelmintics in vitro. The preparations of larvae for cultivation were that the artificially-hatched $L_2$ incubated the embryonated eggs of Ascaris suum in 0.1% formalin solution at $25^{\circ}C$ for 28 days and the rat-derived larvae($L_3$) recovered from the lung of rat infected with the embryonated eggs of Ascaris suum on 7 days after infection(DAI). The cultivation for development of Ascaris suum from the embryonated eggs($L_2$) and the rat-derived larvae($L_3$) for 14 days in RPMI medium 1640(with 5% bovine calf serum) were as follows : 1. The sizes of the liberated larvae($L_2$) which were artificially hatched from embryonated eggs with glass beads(diameter 5mm) were $190{\sim}250{\mu}m$ on 1 days in culture(DIC). The second-stage larvae were molted into third-stage larvae(early $L_3$; $250{\sim}300{\mu}m$) and the features of these larvae were first observed such as cephalic cuticle, esophageal lumen and anus etc. on 5 DIC and the sizes of late third-stage larvae were $250{\sim}450{\mu}m$ on 10 DIC. The sizes of early fourth-stage larvae($L_4$) were $500{\sim}700{\mu}m$ and the features of these larvae were more pronounced in internal organs on 15 DIC. 2. The sizes of third-stage larvae($L_3$) recovered from the lung of rats were $1,340{\sim}1,370{\mu}m$ and the feartures of cephalic cuticle, esophageal lumen, intestine, rectum, anus were visualized by inverted microscope on 1 DIC. The fourth-stage larva($L_4$) completed by third ecdysis were recognizable and sizes of early fourth-stage larvae were developed as $1,400{\sim}2,200{\mu}m$ on 5 DIC. The sizes of middle fourth-stage of larva were $1,900{\sim}2,300{\mu}m$ and the thickened epithelial rectum was observed on 10 DIC. The rectum and anus of late fourth-stage larva($L_4$ $2,500{\sim}3,200{\mu}m$) had developed completely in RPMI medium 1640 on 15 DIC.

혈청학적 진단을 위한 돼지 편충의 체항원, 배설/분비항원의 분리 및 비교

지차호,이철순,박승준,Jee, Cha-ho,Lee, Chul-soon,Park, Seung-jun 대한수의학회 1999 大韓獸醫學會誌 Vol.39 No.1

Swine whipworm(Trichuris suis) is cosmopolitan nematode which can cause serious pathology in immature stage(larva2~larva5) of infected pigs, such as anorexia, diarrhea, anemia, and death in heavy infections. In this larval stages, it is very difficult to diagnose the infection of whipworm and to differentiate from other common swine gastrointestinal disorders such as 21 day scours which are associated with TGE virus, rota virus, coccidium, and the stress of weaning. In this experiment, the isolated antigens of Trichuris spp. were carried out to examine the structure and specificity of antigens and to select the reasonable antigens which would be used in serological diagnosis by electrophoresis, Western blotting, ELISA. The results of this experiment were as follows; 1. The common fractions of each Trichuris suis antigen were identified 28,32,45, 80kDa by SDS-PAGE with silver stain and four major fractions could be detected in positive swine sera by Western blot analysis. 2. The OD(optical density) values of somatic and excretory-secretory antigens which were reacted against positive(negative) sera from pigs infected with Trichuris suis by ELISA reader were; 1) OD values($mean{\pm}SD$) of adult somatic antigen against positive(negative) sera were $0.30{\pm}0.12(0.09{\pm}0.006)$ and third-stage larva of somatic antigen were $0.28{\pm}0.038(0.10{\pm}0.005)$. And OD values of excretory-secretory antigens of adult and third-stage larva were $0.24{\pm}0.031(0.11{\pm}0.005)$ and $0.08{\pm}0.013(0.10{\pm}0.003)$, respectively. 2) OD values of adult somatic, larval somatic antigen and adult excretory-secretory antigen response to positive sera were significantly (p<0.01) associated with negative swine sera. And the Cut-off OD values(minimum positive value) were determined to be mean negative value plus 3 SD that would minimized the risk of false positives. 3. The OD values of somatic antigens of T suis and T vulpis against swine positive(negative) sera were $0.30{\pm}0.120(0.09{\pm}0.006)$ and $0.25{\pm}0.141(0.09{\pm}0.003)$. These data mean that the somatic antigens of T suis and T vulpis were able to diagnose T vulpis infection in dogs as well as T suis infection in pigs. These results suggest that somatic antigen of third-stage larva and excretory-secretory antigen of adult T suis could be used the diagnostic antigen by serological test(ELISA) in immature Trichuris spp. infection.

연충감염우에 있어서 충란검사(蟲卵檢査)에 의한 구충효능(驅蟲效能) 평가방법(評價方法)에 관한 연구(硏究)

지차호,장두환,윤희정,Jee, Cha-ho,Jang, Du-hwan,Youn, Hi-jeong 대한수의학회 1984 大韓獸醫學會誌 Vol.24 No.1

In the evaluation of anthelmintic efficacy by fecal egg counts for cattle naturally infected with helminths, the reasonable technique of fecal egg counts and the reliable guidelines were determined as follows; 1. Modified technique of Harashigeru and Kim's sedimental tube was the most reasonable in fluke egg counts. 2. Universal egg counting technique was preciser than McMaster egg counting technique and was lower in coefficient variation. 3. Fecal egg counts of pretreatment should be carried out twice and mean of the epg should be calculated. 4. Fecal egg counts of posttreatment should be carried out 3 times and established at suitable intervals in consideration of anthelmintic mechanism and withdrawal days of anthelmintics. 5. If nematodal eggs are not found by UECT in posttreatment, direct flotation method should be carried out. And if positive, this epg was calculated at 20 (factor of UECT; $40{\times}1/2$).

돼지 飼料添加劑로써 Fenbendazole의 驅蟲效能評價

池次昊 충북대학교 농업과학기술연구소 1993 農業科學硏究 Vol.10 No.2

The anthelmintic and preventive efficacy of fenbendazloe 4 ppm as feed additive in swine which were naturally infected with and artificially infected A. suum, T. suis and Oesophagostomum spp. during the 60 consecutive days was as follows ; 1.Anthelmintic efficacy of fenbendazole in treatment of pigs which were naturally infected with A. suum, T. suis and Oesphagostomum spp. was 98.73%, 100%, and 100%, respectively. 2.Preventive efficacy of fenbendazole to the establishment of A. suum, T. suis and Oesophagostomum spp., which were artificially infected, was 100%, 96.72%, and 97.83%, respectively. 3.No adverse reactions were observed in pigs treated with fenbendazole 4 ppm in-feed during the experiment. 4.Feed conversion ratio of medicated fenbendazole 4 ppm was better than that of non-medicated control and the weight gain of medicated group was 1.68kg higher than that of non-medicated group. The method of dose, fenbendazole 4 ppd as feed additive during the 60 consecutive days, has very good anthelmintic and preventive efficacy against 3 intestinal nematodes. The marked egg reduction was detected on 10th-30th day of trial by fecal egg conuts. The method of dose in this experiment was the first case in Korea, so the dose of fenbendazole 4ppm has has very good efficacy against A. suum, T. suis and Oesophagestomum spp.. But it is thought that the anthelmintic and preventive efficacy may be lower at the concentration of 4 ppm after some years, because the resistant species of gastrointesinal nematodes against the low level of feed additive anthelmintics would appear at pigsty which is saw-dust fermentation floow system in Korea. But the resistant strains of intestinal nematodes against anthelmintics were not discovered in Korean pig farms. The dose of fenbendazole 4ppm in feed would be reasonable in pigpen of pregnant sow and growing piglets, especially in pigpen of saw-dust fermention floor system in Korea.

가금티푸스 예방을 위한 진드기(옴) 구제방안

지차호 대한양계협회 2001 월간 양계 Vol.33 No.6

人工唾液이 第1胃內 醱酵 및 纖毛蟲의 分布에 미치는 影響

池次昊 충북대학교 농업과학기술연구소 1991 農業科學硏究 Vol.9 No.2

Effects of the experimental artificial saliva on the fiber digestibility, the fermentation values, the population dynamics of the cellulolytic ciliates and bacteria in the rumen of the rams were investigated. Application of the experimental artificial saliva to Holstein cows was also carried out. The results obtained were summarized as follows ; The fiber digestibility and the values of lumen fermentation in the rams were remarkably improved in the treatment group of experimental artificial saliva formulated with sodium bicarbonate(55%), monoammonium phosphate(35%), stabilized rumen extract(2%), sodium chloride(3%), potassium chloride(4%), magnesium chloride(0.5%) and calcium chloride(0.5%) base. The digestibility of cellulose, neutral detergent fiber and acid detergent fiber was significantly improved(p<0.05), whereas, the highly significant increase of the total volatile fatty acid and the soluble phosphorus in the rumen was recognized(p<0.01). Population density of the larger cellulolytic ciliates measured as high as 6.6 times at the 1st week, 16.3 times at the 2nd week, 14.4 times at the 3rd week, and 11.2 times at the 4th week after treatment, respestively, in comparision to the control. Multiplication of the cellulolytic Clostridium sp.(Korean Collection for Type Culture 8440) was also noticed with the level of 1.4×105/ml at the day 12 after treatment. Fractions of the larger lumen ciliates in the treated rams were observed between 60 and 90 μm on the graphs obtained by the Coulter counter channelyzer and it was regared that this type of graphs was applicable to the indirect method to count the rumen ciliates and useful to detect as an indicator of the degree in rumen fermentation. The major species of the larger rumen ciliates in the treated rams were belong to the genera of following order ; Diploplastrom, Polyplastron and Isotricha. The increased milk priductions were noticed as much as 4.5 and 3.3kg/head/day at the 4th and 6th week after treatment, respestively, in the field trial to Holstein cows fed with the specially formulated diet of the experimental artificial saliva and KCTC 8440. Key Words : artifcial saliva, cellulolytic ciliate. cellulolytic bacteria(KCTC 8440), rumen fermentation, ruminant

돼지질병의 실험실진단

지차호,Ji, Cha-Ho 대한양돈협회 1984 養豚 Vol.6 No.10

人工唾液이 第1胃內 醱酵 및 纖毛蟲의 分布에 미치는 影響

지차호 忠北大學校 農業科學硏究所 1992 農業科學硏究 Vol.10 No.2