Considerable attention has been focused on the cryopreservation of semen and estrus induction in dog, as consequence of poor productivity caused by long anestrus period, in order to enhance the productivity of youngs and to preserve the breeds. The objectives of this study were to develop a treatment protocol for estrus induction. Fifty infertilie dogs (age 2∼3 years) were selected for the study and divided into three different estrus induction treatment groups. Group 1: dogs (n=15) were given clomifane (0.1 mg/kg) orally for five days at 12 hr intervals. Group 2: dogs (n=15) were given bromocriptine (50 /kg) orally for five days at 12 hr intervals, followed by single injection intravenously of 500 IU GnRH (Group 3, n=20) when pro-estrus occurred. After being treated, the dogs were evaluated for the rates of estrus induction and time interval lapses from treatment to beginning of the pro-estrus. The estrus induction rates were significantly (P<0.05) higher in both group 2 (9/15, 73.3%) and group 3 (16/20, 80.0%) than that of group 1 (9/15, 60.0%), but did not differ in the groups 2 and 3. No differences were observed in the time interval lapses from treatment to beginning of the pro-estrus in group 2 (7.7 1.2 days) and group 3 (6.9 2.0 days), but significantly (P<0.05) shorter than that of group 1 (9.5 2.1 days). In conclusion, the estrus induction rate of dogs treated with a combination of GnRH and bromocriptine was more effective than use of clomifene or bromocriptine only.

개의 인공수정에 사용할 정자의 보존방법을 확립하기 위하여, 동결속도와 응해 온도를 설정하여 적절한 동결방법을 정립하고자 본 실험을 실시하여 다음과 같은 결과를 얻었다. 동결의 방법에 있어서는 액체질소의 표면으로부터 17 cm 높이에서 동결하는 -3/min의 동결속도로 실시하여 37에서 2분간 응해하는 방법이 가장 좋은 결과를 보였다. 생존성과 운동성에 있어서의 차이는 없지만 첨체의 intact한 비율은 약간 낮은 결과를 보였으며, 이의 보완을 위해, 액

Artificial insemination (AI) with frozen or cooled semen is widely used in commercial fields of cattle and pig. Little is known about characteristics of canine sperm after freezing or cooling. For both practical and commercial goal, the canine semen treated with cooling and freezing should be carried out to exam the fundamentals, including sperm motility, survivability and fertilizing capacity. The aim of this study, thus, was to identify the effects of extended exposure to 4 on canine semen by motility, survivability, acrosomal changes following different duration. Fifteen ejaculates collected by digital manipulation twice per week from 3 dogs (Shih-Tzu) were divided to 16 aliquots after adding Tris-egg yolk (TE) buffer formulated by our laboratory, and cooled from 37 to 4, by ramp rate of 0.6/min. Each sample was evaluated by their motility, survivability and the acrosomal status at 0h (control), 2h, 12h and 1 d~10 d, respectively. The motility of spermatozoa was graded to 6 levels using the modified method of Seager. The survivability of sperm was assessed using an epifluorescence microscope after Fert/Light (Mole-cular Probes Inc.) staining. To estimate the proportion of the spermatozoa of intact acrosome, 200 spermatozoa were assessed in randomly selected fields, using epifluorescence microscope after FITC/PSA (Sigma) staining. At 2 h after cooling, the motility of most spermatozoa were assessed to be grade 0 and 1. At 12 h, high number of sperm were in grade 0 to 1, however, it was significantly (P<0.05) lower than that of 2 h. From 1 d to 4 d, ~50% of sperm was assessed to grade 0 to 1. On day 7, a little sperm were in grade 0 to 1. No sperm showed motility on day 10. Sperm motility was rapidly reduced by the percent of 10% of grade 0 to 1. From 2 h to 6 h, the number of live sperm was 90% and the sperm chilled for 10 days lived>50%. Acrosomal intact of spermatozoa exposed to 4 for 2 h was 51%, supposed the sperm of control was 100%. Our results suggest that 1) this is easy to transfer and preservation for short periods 2) AI can be used by semen chilled for 6-Day.

소와 견에 있어서 Unopette가 정자의 형태학적 검사 및 정자농도의 검사를 위하여 사용될 수 있는가를 알아 보기 위하여본 연구를 수행하였다. 소정액 및 견정액을 Unopette에 희석한 후 3-5에 보존하면서 시간경과에 따라서 위상차현미경하에서 관찰하여 다음과 같은 결과를 얻었다. 1. Unopette를 사용하여 관찰한 정자는 48시간까지는 hematoxylin-eosin을 사용하여 정자보다는 높은 정상정자율을 나타내넜다. 2. Unopette를 사

This study was designed to evaluate the sensory characteristics and consumer preference of dog meat foods as compared with beef ones. The sensory evaluation was conducted by a 10-member trained panel and 109 persons ranging in age from 23 to 59 participated in the consumer research. The results were summarized as follows: 1. The sensory characteristics. 1) In case of the meats boiled in water, it did not show any significant differences between dog and cow's meat in color as well as off-flavor. On the contrary, the other characteristics such as odor, tenderness, juiciness and oiliness of dog meat were evaluated stronger than those of beef. 2) when the meats were cooked as Tang (a kind of soup), the dog meat did not show any significant differences from beef not only in color and off-flavor but also in odor. 2. The consumer preference. 1) It appeared that consumers somewhat preferred beef Tang to dog meat Tang. However, they rated dog meat Tang as the 'neither liked nor disliked' food on an average. 2) Male consumers showed higher preference than female did for the dog meat tang. On the overall, dog meat foods are regarded to have some desirable sensory characteristics and can be acceptable to most people.

In this study, the kinds of Dog Meat Cooking, side dishes, ingredients, seasonings and recipes were surveyed in 21 Dog Meat Cooking restaurants in Korea from July to August of 1989. 1. Actually, there were four Dog Meat Cooking recipe. Tang (soup:湯) has been served in 20 (95.2%) restaurants and Sukyuk(boiled in water:熟肉) in 19(90.5%) ones. Junkol(boiling vegetables and meat with seasoning:煎骨) and Muchim(sauteed with seasoning:무침) has been done in 16(76.2%) and 10(47.6%) restaurants, respectively. 2. According to the frequencies, the main seasonings when served were roasted perillar powder (95.2%), soybean paste (95.2%), vinegar(81.0%), Dadegi (mixed much red pepper powder, minced ginger, minced garlic, minced onion and black pepper powder, 66.7%), pepper(61.9%), salt(61.9%), salt(61.9%), minced ginger(57.1%), minced garlic(57.1%) and prepared mustard(38.1%). 3. The number of side dishes were 26. Among vegetables, green pepper(90.5%), sliced garlic(81.0%) were served. Chinese cabbage(61.9%) and Kagtuki(42.9%) out of Kimchi and leek salad(28.6%) were also served. 4. The total 17 ingredients were used in Dog Meat Cooking. The major one were white onion (100%), perillar leaf(72.2%), leek(66.6%) and parsley(47.2%).

1670년부터 1943년까지의 문헌 16권을 통하여 견육(犬肉)요리의 종류, 조리법, 양념, 부재료를 조사한 것은 다음과 같다. 1) 견육(犬肉)요리는 14종류로 분류되며, 연대순으로 보면 「찌는법」(증(蒸)) 20회(40%), 「견육순대」 1회(2%), 「견육꽂이구이느르미」 1회(2%), 「견육느르미」 1회(2%), 「국(갱(羹),탕(湯))」 12회(24%), 「익힌 고기 다시 찌는 법」 4회(8%), 「구장」(구장(狗醬)) 4회(8%), 「구적」(구적(狗炙)), 「구장과 젖」(구장(狗醬)과 해), 「구포」(구포(狗脯)), 「구족초」(구족초(狗足炒)),「구이진초」(구이진초(狗彛唇炒)), 「백숙」등은 각각 1회(2%)이며 50회 기록되었다. 빈도면에서는 「찌는 법」 「국」 「익힌 고기 다시 찌는 법」 「구장」의 순으로 「찌는 법」이 주를 이루고 있었다. 2) 견육요리의 전처리에 있어서 씻는 과정은 내장만 씻고 고기는 씻지 않는 것(52.6%)이 전부를 씻는 것(26.3%)보다 많았고, 내장을 많이(42.1%) 이용하였으며 익히는 과정에서는 거의 쪄낸 후(84.2%) 나름대로의 요리를 만들었고 삶어서 이용한 것은(9.0%) 거의 없었다. 3) 전체 견육요리에 이용된 양념은 22가지이며 그 중 많이 이용된 것은 유장(42.1%), 참깨가루(39.4%), 후추가루(36.8%), 식초(36.8%), 간장(28.9%), 고춧가루(26.3%), 참기름(23.6%), 만초가루(23.6%), 천초가루(23.6%) 등이고, 부재료는 5가지이며 그 중 많이 이용된 것은 파(파의 흰부분과 합함 34.2%)와 미나리(21.0%)이다.

The present study aimed to examine the effect of dietary Ptecticu tenebrifer powder mixtures as pet dogfeed ingredients on crude fat and ash digestibility. Three groups of feeds Feed A, Feed B, and Feed C supplied from three farms were fed to a total of 45 dogs. The dietary Ptecticu tenebrifer powder mixture were prepared by mixing 25 g of Ptecticu tenebrifer powder with 100 g of canned food. Feed A, Feed B, and Feed C containing dietary Ptecticu tenebrifer powder mixtures were fed to 15 dogs of each breed of bichon, poodle, and chihuahua that were divided into three groups following a completely randomized design. For measuring the crude fat and crude ash digestibility, manure of each dog breed from each group were collected. Crude fat digestibility was not statistically significant among the dog breeds fed with feed C (p>0.05), but overall there was a statistical difference between the feed and the group by dog breed (p<0.05). In terms of crude ash digestibility, the three types of feed showed differences with respect to dog breeds (p<0.05). However, the group with no significant difference was observed in Feed B by dog breed (p>0.05). In conclusion, feeding Ptecticu tenebrifer powder mixture to dog breeds had no positive effect on the crude fat and ash digestibility and can be used as pet dogfeed ingredients.

A rapid and simple method for the quantitative determination of volatile fatty acids (VFAs; propionic acid, n-butyric acid, i-valeric acid and n-valeric acid) and indoles (phenol, p-cresol, 4-ethyl phenol, indole and skatole) in pig slurry and dog excrement using solid-phase micro-extraction (SPME) coupled to gas chromatography was evaluated. 50/30 ㎛ DVB/CAR/PDMS (Divinylbenzene/Carboxen/Polydimethylsiloxane) fiber was used to extract the target compounds in aqueous media. Sample amount and adsorption time was standardized for the routine analysis. Detection limits were from 0.11 to 0.15 ㎍/L for VFAs and from 0.12 to 0.28 ㎍/L for indoles and the correlations observed (R2) were 0.975～1.000. This method was applied to the pig slurry, fertilizer, compost and dog excrement. In nearly all cases, the indoles were detected in concentrations of higher than their limits of detection (DOLs). But the VFAs in swine manure were below their DOLs.

Cleft palates with or without cleft lip is one of the most common congenital craniofacial defects in dogs. It has been reported that monogenic autosomal recessive inheritance caused this defect in this species. However, here, we aimed to report cleft palate in a cloned dog. A fibroblast cell line was established from skin tissues of an eight-year-old German shepherd dog. Blood was collected from oocyte donor dogs, and serum progesterone concentration was measured by chemiluminescence enzyme immunoassay method. Ovulation was determined when serum progesterone results reached 5-10 ng/ml, and in vivo matured oocytes were collected surgically about 72 hr after ovulation. Donor cells were cultured with Dulbecco’s modified Eagle medium supplemented with 10% (v/v) fetal bovine serum until confluence. An in vivo matured oocyte was enucleated, and a donor cell was injected into the perivitelline space. The oocyte-cell couplet was electrically fused, and chemically activated. Reconstructed embryos were transferred to an oviduct of a recipient. Pregnancy diagnosis was performed 27 days after the embryo transfer, and ultrasonography of fetal heart beat, and rectal temperature and serum progesterone value of recipient was monitored until the day of delivery. Microsatellite analysis was performed using genomic DNA of cell donor, clones, and oocyte donors. As results, a total of 74 cloned embryos were transferred to five recipients, and one recipient diagnosed as pregnant with two fetuses by ultrasonography and radiology. Caesarean section was performed on day 58 after embryo transfer due to a decreased heart beat of a fetus, which was lower than 180. Two cloned puppies with 640g and 320g of birth weight were delivered safety, but the small one was born with a cleft palate. Microsatellite analysis results of both clones were identical with the cell donor. Cleft palate of the clone was surgically corrected on day 40 after birth. To our knowledge, there has been no report about cleft palate in cloned dogs, and also, no report about clones with different phenotype of cleft palate in dogs. Therefore, this study can give a clue of cleft palate in dogs, which might not be a genetic cause. Further studies about aberrant epigenetic reprogramming in those clones are needed.

개에서 이물, 신생물, 유문동비후, 위수술, 전해질불균형, 위확장성 염전 등에 의한 비정상적인 위 배출시간은 임상에서 소화기 질환으로 중요하다. 그러므로 위장관 운동이상에 대한 정확한 진단을 위하여 정상적인 위장관 운동시간에대한 자료가 필요하다. 이 연구의 목적은 개에서 기존의 BIPS를 이용하는 검사 방식이 아닌 국내에서 개발한 방사선비투과성 Kolomark를 이용한 위배출시간 검사에 대한 임상적 유용성에 대한 기초자료를 제공하기 위한 것이다.Beagle 9마리가 이번 실험에 사용되었으며 평균 체중 평균 10.3kg이며 평균 2.5살이었다. 검사를 위해 12시간 금식을시행하였으며 화학적 보정은 하지 않은 채 검사 직전 하루 사료급여량의 25%용량에 Kolomark 1개의 capsule과 함께 급여하였고 2, 4, 8, 12시간 때 Ventrodorsal, Right lateral자세로 촬영하였다. 관심판독부분은 분문에서 위유문부까지의 위장 전체를 관찰하였으며 분석방법으로는 각 시간대별로 위장 내에 남아있는 Kolomark를 카운트하여 비모수검정인 Friedman 검정방법을 이용하여 P값이 0.05 미만인 경우를 유의한 것으로 판정하였다. 구강으로 Kolomark를 섭식후 위에서 소장으로 완전히 Kolomark가 빠져나가는데 걸리는 평균시간은 7.55시간이었다. 이번 연구에서 성숙한 개에서 음식물 투여 후에 나타나는 위장관 통과시간을 Kolomark를 이용하여 정상적인 위장관 운동시간에 대한 기초자료가되리라 판단된다.

The canine major histocompatibility complex (MHC) is referred to dog leukocyte antigens (DLA), which is known to be the most polymorphic genetic system in canine species. Many cloned dogs have been produced since Snuppy, first cloned dog, there was no research about genetic identity of MHC among cloned animals. Recently in Lee’s group, two non-transgenic cloned beagles (BG1, 2) were produced by somatic cell nuclear transfer (SCNT) using fetal fibroblast (BF). Also, four transgenic cloned beagles (Ruppy 1-3, 5) were generated using transgenic BF transfected with Red fluorescent protein (RFP) gene. We hypothesize that non-transgenic (BG1, 2) and transgenic (Ruppy 1-3, 5) cloned beagles derived from identical donor cells have the same immunological genetic characteristic except for RFP gene insertion in the genome. Thus, the aim of this study is to confirm the immunological identity of DLA class II in cloned beagles produced using same nuclear donor cell. Genomic DNA was extracted from blood of BG1, BG2, Ruppy 1, 2, 3 and 5. Genomic DNA of normal two control beagle, no correlation with BF was also investigated for rulling out the possibility that beagles were inbred. Forward and reverse primers used for DLA-DQA1 and DQB1 respectively were DQAF: 5’-TAAGGTTCTTTTCTCCCTCT-3’ and DQAR: 5’-GGACAGATTCAGTGAAGAGA-3’ DQBR:5’-CTCACTGGCCCGGCTGTCTC-3’ and DQBR: 5’-CACCTCGC CGCTGCAACGTG-3’. Polymerase Chain Reaction (PCR) products were purified, sequenced directly using the Big Dye Terminator kit. Sequencing analysis was performed on an automated 3730xl DNA analyzer. In experiment 1, sequence of DLA-DQ alpha 1 (DQA1) and DLA-DQ beta 1 (DQB1) exon 2, hypervariabel region, was compared in BG1 and BG2. Experiment 2 also compared the sequence of DQA1 and DQB1 among Ruppy 1, 2, 3 and 5. Experimental 3 compared sequence of DQA1 and DQB1 among all cloned dogs (BG1, BG2 and Ruppy 1-3, 5). As a result, BG1 and BG2 have same allele for DQA1 and DQB1 as we expected. They share DQA1*00101 and DQB1*02901 in experiment 1. In experiment 2, Ruppy 1, 2, 3 and 5 also have identical DQA1*00101 and DQB1*02901 allele. No discrimination between transgenic dogs and cloned dogs was seen in DQA1 and DQB1 Allele in experiment 3. DQA1, DQB1 allele was identified as *00101 and *02901 in all dogs. We provided the allele identity of DQA1and DQB1 in cloned beagles, which can be used as preliminary data for immunological related studies. In conclusion, transgenic cloned dogs despite of red fluorescent protein genes being inserted in their nuclear DNA were immunologically compatible with non-transgenic cloned dogs. We demonstrated that cloned beagles produced using identical nuclear donor were immunologically compatible.

Controllable transgenic expression systems in transgenic animal model are valuable to the development of therapeutic approaches in human medical fields. The aim of this study was to 1) produce a transgenic cloned dog using inducible tetracycline vector system, and 2) investigate whether the transgenic cloned dog could be induced the transgene expression using doxycycline (Doxy). Canine fetal fibroblasts were infected with retroviral vectors designed to express the enhanced green fluorescent protein (eGFP) gene under the control of tetracycline-inducible promoter. For somatic cell nuclear transfer (SCNT), nucleus of an in vivo matured oocyte was removed and an eGFP expressed cell cultured with 1 ㎍/㎖ of Doxy was injected. After electrical fusion and chemical activation, the reconstructed embryos were transferred to a recipient and pregnancy diagnosis was performed by ultrasonography. Experiment I evaluated the mean fluorescence intensity (MFI) of infected cells while the cells were cultured in the presence of 1 ㎍/㎖ of Doxy for 5 days, and then in the absence of Doxy for 7 days using fluorescence-activated cell sorter. Experiment II was designed to produce an eGFP controllable transgenic cloned dog via SCNT. For verification of transgenic dog, experiment III was performed Southern Blot analysis and observation in vivo regulation of eGFP expression in the cloned dog treated with 100 ㎎/㎏ of Doxy every 2 days for 2 weeks under ultraviolet light. In experiment IV, western blot was used to detect eGFP increase and decrease in skin tissues of transgenic dog under the presence or absence of Doxy. In the results of experiment I, the MFI for infected cells was rapidly increased to approximately 42.3 times after 3 day-treatment compared to pre-treatment and quickly decreased 3 days after ceasing the treatment. In experiment II, a total of 203 embryos were transferred to nine recipients and three pregnant delivered three pups (Tet-on eGFP 0, Tet-on eGFP 1, and Tet-on eGFP 2) by C-sec and Tet-on eGFP 2 among them is still alive. All cloned pups were genetically identical to the donor cell. Tet-on eGFP 2 showed an apparent in vivo eGFP expression on her body after Doxy administration in experiment III. The result of Sothern blotting showed that the transgene insertion was detected from the three cloned dogs and all organs of Tet-on eGFP 1. Experiment IV indicated that a robust eGFP expression in skin tissue of Tet-on eGFP 2 rapidly increased after Doxy treatment and gradually decreased to basal level on 9 weeks after ceasing the treatment. In conclusion, we report here for the first time an inducible transgenic system in canine species and it can stably induce the transgene expression at intended time. This study has demonstrated the capacity to generate transgenic model dog which could regulate the transgene and it would contribute to human medical research fields.

The cloning of canids was succeeded in 2005, several years after the birth of Dolly the sheep and also after the cloning of numerous other laboratory and farm animal species. The delay of successful somatic cell nuclear transfer (SCNT)was due to the unique reproductive characteristics of the female dogin comparison to other domestic mammals, such as ovulation of immature canine oocyte and a requirement of 25 days for the completion of meiosis within the oviduct (Holst & Phemister, 1971). When the technology for the recovery of in vivo matured oocyte was established, the application of cloning also became possible and cloned dog offspring were obtained. This report summarizes the progress of technical procedures that are required for cloning canids and the application of this technique. The first cloned dog, Snuppy, was achieved using an in vivo-matured oocyte which was enucleated and transferred with an adult skin cell of male Afghan hound. After establishment of a criterion of well-matured oocyte for the improvement of SCNT efficiency, we obtained three cloned female Afghan hound and a toy poodle cloned from 14 year-old aged Poodle using SCNT through this factor. To date, cloned dogs appeared to be normal and those that have reached puberty have been confirmed to be fertile. Through application of canine SCNT technique, first, we demonstrated that SNCT is useful for conserving the breed of endangered animal from extinction through cloning of endangered gray wolves using inter-species SCNT and keeping the pure pedigree through the cloning of Sapsaree, a Korean natural monument. Secondly, we showed possibility of human disease model cloned dog and transgenic cloned dog production through cloning of red fluorescent protein expressing dog. Finally, SCNT can be used for the propagation of valuable genotypes for making elite seed stock and pet dog. In summary, dog cloning is a reproducible technique that offers the opportunity to preserve valuable genetics and a potential step towards the production of gene targeted transgenic cloned dogs for the study of human diseases.

We have studied the effects of dampening papers (Dampened paper, Newspaper, Korean paper, Flower sheet), silica gel (30g, 60g) and temperature (20℃, 25℃) on color changes of Erythronium japonicum. In the treatment of 20℃, color changes were low in treatments with silica gel rather than in a treatment of dampening papers. In particular, newspaper and Korean paper showed much less changes in colors by the combination treatments with 30g of silica gel. Likewise, in the treatment of 25℃, color changes were low in combination treatments with silica gel rather than in a alone treatment of dampening papers. For the combination treatment with 30g of silica gel, low color changes were shown in the divisions of newspaper and Korean paper, while for the combination treatment with 60g of silica gel, low color changes were shown in the those of Korean paper and dampened paper.