화학합성 농약을 대체하기 위한 친환경 방제법 개발을 위해 1, 2, 4%의 도꼬마리 열매 수용성 추출물을 이용하여 팥바구미 살충력을 실험하였다. 실험한 추출 농도에서 곤충 치사율, 기피성, 산란율과 성충 우화 및 팥의 피해 억제에 효과를 보여 주었다. 4%의 추출물은 처리 후 2일 후에26%의 치사율과 처리 후 3시간에 53.3%의 기피율을 각각 보였다. 또한 4%의 추출물을 팥과 섞었을 때 가장 낮은 산란율(113.7/암컷)과, 성충 우화율이 가장 많이 억제(37.0%)되었고 종자 손상을 가장 적게(42.3%) 받았다. 이러한 결과는 도꼬마리 추출물이 저장곡물을 팥바구미로부터 보호하는 종합적 방제수단으로 사용될 수 있다.

Semen can be divided into two parts. One is cellular part which contains sperms the other is liquid part which is called by seminal plasma. The seminal plasma is a nutritive and protective medium for the sperms. Fructose, which is major energy source, is supplied to sperms swim to female oocyte. Alkalic property protects sperms from hostile environment of female reproductive organ. Also, seminal plasma induces tolerance to preexisted immune cells, and changes intra‐uterine environment to better conditions for fertilized embryos to implant. However, the effects of seminal plasma in in vitro culture of fertilized embryos are unclear. Second fraction of fresh semen was obtained from a normal farm pig. The semen was centrifuged to remove sperms, and then supernatant was filtrated. The filtered seminal plasma was stored in — 30℃. In this study, electrically activated and chemically activated porcine embryos were employed to investigate the developmental rate after 2 hours treatment of none, 0.1%, 0.5%, and 1% seminal plasma in culture media by two days of activation. Both electrically and chemically activated embryos, cleavage rate and cell numbers of blastocysts were not significant difference within four groups. Blastocyst formation rate of electrically activated embryos also did not show significant difference within any groups. However 0.1% seminal plasma treatment group showed significantly increase of blastocyst formation rate in chemically activated group (None; 24.8%, 0.1%; 31.7%, 0.5%; 19.4, and 1%; 16.5%, respectively. p<0.05).

In the present study, we investigated the effect of porcine follicular fluid (PFF) concentration (10% vs. 1%) and protein-free media (PFF 0%) on maturation of porcine oocytes in vitro and analysed difference in gene expression in resulting blastocysts following parthenogenetic activation. Three groups were tested; 1) 10% PFF: Tissue culture medium (TCM) 199+10% PFF; 2) 1% PFF: TCM 199+1% PFF; and 3) 0.1% PVA: TCM 199+0.1 PVA. Cumulus-oocyte-complexes were cultured in the respective media containing gonadotrophin (1 ug/ml), epidermal growth factor (10 ng/ml), cystein (0.57 mM), sodium pyruvate (0.91 mM), insulin (5 ug/ml), 9-cis retinoic acid (5 nM) for 20～22 h and then without hormonal supplements for an additional 20-22 h. Data was analyzed using statistical analysis system(SAS) program. There was no significant difference in oocyte maturation rate. However, significantly higher (p<0.05) proportions of embryos developed to the blastocyst stage when oocytes were matured in 10% PFF group (45%) than in the 1% PFF group (31.1%). The total cell numbers were not significantly different among groups (52 ± 1.3 vs. 54.6±3.1 vs. 54.4±2.5, respectively). The relative abundance (ratio to beta-actin mRNA) of gene transcripts related to apoptosis in blastocysts was measured by real- time PCR. The expression of anti-apoptotic gene (BclxL) was up-regulated and the expression of pro-apoptotic gene (Bax) was down-regulated in 10% PFF group than in the other groups. Therefore, it can be concluded that supplementation of 10% PFF during in vitro maturation improves embryo development by reduction of apoptosis. * This study was supported by IPET (#311011-05-1-SB010), RNL Bio (#550-20120006), MKE (#10033839-2011-13), Institute for Veterinary Science, the BK21 program and TS Corporation.

Embryo transfer (ET) is the final procedure for getting pregnancy through assisted reproductive technology such as IVF (in vitro fertilization), SCNT (somatic cell nuclear transfer). In our laboratory, the porcine cloned embryos loaded in ET medium are carried for 3 hours by portable incubator because of the great distance from the laboratory to the experimental farm. Thus, before transferring into recipient, porcine cloned embryos are exposed in vitro condition for long time. Medium which is used in this process is the TALP (Tyrode’s medium supplemented with 10 mM HEPES), but it includes little nutrients for embryo. Thus, the aim of this study is to determine whether ET media containing nutrients affect the in vitro development of embryos compared to TALP. For the experiment, porcine zygote medium (PZM)-5 which has amino acids for developing embryo was chosen as ET medium containing nutrients, added 10 mM Hepes as PZM-5 does not contain buffering system. For experiment, we carried out parthenogenesis through a chemical method using Thi/DTT. Parthenogenetic embryos were cultured in PZM-5 for 2 days, and then they were randomly divided into two group; loaded in a straw with TALP or PZM-5-Hepes, respectively. They were stored in a portable incubator for 3 hours to simulate the time consumed in ET, thereafter embryos in both TALP and PZM-5-Hepes groups were respectively cultured in PZM-5 for additional 5 days. All experiments were repeated 5 times. In result, blastocyst formation rate were 22.46%±1.47 and 23.17%± 2.13, respectively and total cell number were 32.9±2.22 and 37.09±2.18, respectively. There is no significant difference between TALP and PZM-5-Hepes groups. * Further study will investigate effect of PZM-5-Hepes on in vivo development of porcine cloned embryo. This study was supported by IPET (#311011-05-1-SB010), RNL Bio (#550-20120006), Institute for Veterinary Science, the BK21 program and TS Corporation.

Adipose tissue-derived mesenchymal stem cells (ASCs) are very interesting in several laboratory animals and humans because they are easy to harvest and expand to generate millions of cells from a small quantity of fat. ASCs are known as useful materials for clinical applications in human cell therapy and as a donor cell in somatic cell nuclear transfer (SCNT). Here, we investigated if 1) minipig ASCs can be isolated, self-renewed and differentiated into multiple tissue lineages, 2) ASCs can be a suitable donor cell type for generation of cloned pig. In order to isolate ASC, adipose tissues were collected from inguinal region of a 6-year-old female minipig. The ASCs were attached to the culture dish with a fibroblast-like morphology. They expressed cell-surface marker characteristics of stem cell, underwent osteogenic, adipogenic, myogenic, neurogenic and chondrogenic differentiation when exposed to specific differentiation-inducing conditions. To investigate its potential as donor cell for cloning, we respectively carried out SCNT using ASC, adult skin fibroblast (ASF) and fetal fibroblast (FF) derived from same minipig. The ratio of blastocysts to 2-cell embryos and total cell number of blastocysts were monitored as experimental parameters. In results, cleavage and developmental competence to blastocysts rate showed no significant difference among the three groups. On the other hand, total cell numbers of blastocysts derived from ASC and FF were significantly higher than in ASF (89±7.9 and 105±5.5 vs. 57.5±5.2, respectively). Our results demonstrated that ASC have potential compared to ASF and FF in terms of the in vitro development and blastocyst formation ability. In further study, we will investigate the in vivo developmental ability of ASC as donor cell for pig cloning. * This study was supported by IPET (#311011-05-1-SB010), RNL Bio (#550-20120006), Institute for Veterinary Science, the BK21 program, TS Corporation and Optifarm Solution.

An assessment was made of the biological control potential of mud loaches, Misgurnus mizolepis, toward Culex pipiens molestus, in laboratory condition and septic tank and rainwater storage tank (RST) systems. Results were compared with those of temephos 20% emulsifiable concentrate (EC) and Bacillus thuringiensis israelensis (BTI). In the laboratory tests, all M. mizolepis survived on waters from the settling tank of aerobic septic tank (AST), sump tank of AST, and RST. However, all M. mizolepis died within 3 h after introduction in the settling tank and sump tank waters of anaerobic septic tank (AnAST). Gill or dorsal fin inflammation was detected in the dead mud loaches. M. mizolepis consumed an average of 968–1087, 901–986, and 993–1087 of 1500 third instars of Cx. p. molestus in AST settling tank, AST sump tank, and RST waters, respectively. In the AST and RST systems, predation of Cx. p. molestus by mud loaches at a release rate of 900 larvae/fish resulted in complete mosquito control from the first wk after treatment through the end of the survey period for 16 wk. The average mosquito reduction rates by temephos 20% EC and BTI treatments were 28.6 and 2.1% 2 wk post-treatment, respectively. Mud loaches merit further study as a potential biological control agent for the control of mosquito populations in light of global efforts to reduce the level of highly toxic synthetic insecticides in the aquatic environment.

In haplodiploid sex determination, females are sexually reproduced from fertilized diploid eggs, and males from unfertilized haploid eggs. Haplodiploid sex determination seems simple in that sex depends simply on the ploid level. However, the underlying genetic mechanisms are thought to be much more complicated than expected. Among them, a powerful proposed mechanism is genomic imprinting. All epigenetic on-off systems require target genes, unless the systems target histone proteins on chromosomes. For Hymenoptera, a good candidate target gene in terms of sex determination is known either as feminizer (fem) or transformer (tra) in many insects. These two genes are essential for expressing femaleness. In most Hymenopteran insects, the maternal tra seems to be methylated and consequently not expressed, while the paternally derived tra gene is not methylated. Therefore, a fertilized egg with the paternally derived active tra gene will develop into a functional female. Like all Hymenoptera, ants (Formicidae) have haplodiploid sex determination. In Vollenhovia emeryi, however, queens are produced clonally while workers derive from fertilized eggs. Males are haploid, likewise deriving from fertilized eggs, but only after selective elimination of their maternal genome. Under the conventional genomic imprinting model, we would have expected that the opposite pattern of what is observed in others. Here we present extraordinary sex determination and suggest our hypothesis about genomic imprinting pattern in V. emeryi

Vollenhovia emeryi ant is distinguished by its wing morphology; short winged (SW) and long winged (LW). Its reproduction shows a bizarre genetic caste system distinct from other social hymenopteran insects. Unfertilized eggs undergo genome duplication and develop into clonal gynes. Fertilized eggs develop either into workers or males. The fate of the fertilized eggs is determined whether maternal genome loss (MGL) takes place after fertilization. Eggs with MGL become haploid males with only paternal half of the genome. Without MGL, the eggs become workers with maternal and paternal half of the genome. In this research, we analysed 5 nuclear genes of SW and LW individual ants. Among them, two genes from an SW male are identical to those of LW, and one gene from the SW male seems a variant of LW. The result indicates that SW males are derived from LW colonies. From the genetic relatedness point of view individuals in the same castes are genetically identical. On the other hand, between workers and two reproductives, the relatedness is asymmetrical and there is even no gene sharing between gynes and males. The conventional genetic relatedness by Hamilton is revised under this condition.

The ant species, Vollenhovia emeryi, is distributed in Far East. The species can be divided into two major groups by their wing morphology of reproductives: short-winged and long-winged. A nationwide survey of the species was conducted for analyzing the mitochondrial haplotype diversity and genetic population structure. We collected 91 samples from 40 locations. A total of the 1239 bp partial COI (cytochrome C oxidase 1) region was used for the analyses. We found the total of 21 haplotypes. The mitochondrial haplotypes may correspond to the wing morphology. The genetic population structure examined potential geographic barriers of gene flow such as distance, mountains, rivers and plains which are non-mountain areas to prevent dispersal through mountain range. The result implied that no barriers considered in this study affected differently gene flow. Therefore, the behavioral characteristics of the ant may be the causal constraint of its genetic exchange.

This study was carried out to examine the safety of Coccimuel-S, an anticoccidiol, using male and female ICR mice. Mice were orally administered Coccimuel-S at dose levels of 250, 500, 1,000 and 2,000 / body weight for singledose toxicity test. There were no significant differences in change of body weight between control and all groups treated with Coccimuel-S. Also, general toxic symptoms, clinical signs and mortality were not observed. In the hematological analysis, none of the parameters were affected by Coccimuel-S. This suggests that there are no negative effects on homeostasis and immunity. In blood biochemistry analysis, none of the markers were affected by administration of Coccimuel-S. Similarly, there were no significant effects on markers for liver, kidney, skeletal and heart muscle functions in all treated-groups. No remarkable lesions were detected in these organs with macroscopic examination. Since there were no adverse effects of Coccimuel-S in single oral toxicity tests, even at higher doses than normal, it was concluded that Coccimuel-S could be candidates as a safe anticoccidials for the treatment of poultry coccidiosis.

Minipigs are regarded as one of the most important laboratory animal in that anatomical and physiological properties are similar to human and their reproduction efficiency is relatively higher compared to other large animal species. Particularly, several diseases that cannot be mimicked in rodent models are successfully occurred or induced in pig models therefore it has been interested in a valuable model for human diseases. Pigs are also ‘standard’ species in xenotransplantation research. To maximize experimental outcome using minipigs, establishment and management of proper animal facility, right animal husbandry and control of pathogens are very important. In this review, we summarized several international guidelines related with minipigs published by several companies or governments and discuss optimal conditions for providing informative ideas to the researchers who want to use minipigs in their future studies.

Vollenhovia emeryi chosenica (Wheeler) (Hymenoptera: Myrmicinae) is an ant species frequently found in forests. In nature, two phenotypically distinct forms are found e.g. long winged and short winged. Unlike other hymenopteran insects, the ant is unique in its mode of reproduction. In this species, queens are clonally reproduced from unfertilized eggs. On the other hand, workers develop from fertilized eggs. Strikingly, haploid males are reproduced from fertilized eggs after destroying the maternal half of the genome e.g. maternal genome loss (MGL) consequently only with the paternal half of the genome. We collected the ant colonies nationwide in 2011. In this study, we demonstrate that the ant is infected with Wolbachia, the bacterial reproductive manipulator in various insects. Interestingly, only the long winged morphs seem to be infected. Furthermore, most colonies are mulitple-infected except two colonies collected from Chuncheon and Mt. Deogyu. We will discuss potential interactions among the Wolbachia infection polymorphism and wing morphology, and evolution of clonal reproduction and MGL.