The Varroa mite, Varroa destructor, a parasitic mite that afflicts honey bees, has become increasingly resistant to acaricides like fluvalinate due to its widespread use. The target site insensitivity mechanism, mediated by the L925V/M/I mutations in the voltage-gated sodium channel, plays a major role in resistance. Additionally, cytochrome P450 monooxygenases (Cyp450s) appear to function as a metabolic resistance factor; however, no Cyp450-mediated resistance mechanism has been reported to date. The aim of this study was to identify and characterize Cyp450s associated with fluvalinate resistance. A synergistic bioassay confirmed the involvement of Cyp450s in conferring tolerance or resistance to fluvalinate. Correlation analysis between mortality data and the expression levels of Cyp450 genes led to the identification of several candidates that may play a crucial role in fluvalinate resistance. Analysis of tissue distribution patterns revealed that these genes were most abundantly expressed in the cuticle and synganglion. This suggests that, despite their relatively low expression level, they may play a critical role in protecting the target site from fluvalinate due to its predominant expression in neuronal tissues. Functional analysis, in conjunction with baculovirus expression, demonstrated that fluvalinate has high inhibition rates against the recombinant candidate Cyp450s, suggestive of their strong interaction with fluvalinate. We discussed the potential utilization of their expression levels as a molecular marker for diagnosing metabolic resistance in field-collected Varroa mites.

A 13-year-old mixed dog was referred to the animal medical center, Gyeongsang National University. Lung masses were diagnosed at the left cranial and caudal lobes through diagnostic imaging, and consequently, left pneumonectomy was performed using a self-cutting linear endoscopic stapler. The pulmonary arteries, veins, and bronchus of each lung lobe were sealed and resected at once, and any air leakage or bleeding was not observed after the surgery. Compared to the conventional ligation method, the self-cutting linear endoscopic stapler has the advantage of significantly reducing the operation time and enabling simple and reliable sealing.

To identify and compare the venom components and expression patterns of some bees/wasps, venom gland-specific transcriptome analyses were conducted for 14 Aculeate bees/wasps. Most of the allergens and pain-producing factors showed extremely high expression levels in social wasps, implying that social wasps have evolved to use venom to defend the colony against intruders. Acid phosphatase and tachykinin, which are known as allergens and neurotoxic peptides, were found with high frequencies in the venom glands of solitary wasps. This suggests that solitary wasps might use their venom for catching and preserving prey. In the venom glands of bumblebees, little or no transcripts of major allergens or pain producing factors were identified, implying that bumblebees venoms are relatively less toxic than those of social or solitary wasps. Taken together, the differential expression patterns of venom genes in some Aculeate bees/wasps implies that bees/wasps have unique groups of highly expressed venom components, which appear to have evolved in response to both ecological and behavioral influences.

Mosquitoes are primary medical insect pests due to their diseases transmission as vectors. In Korea, the insecticide-resistant populations of disease vector mosquito species, such as Anopheles sinensis, Culex pipiens and Culex tritaeniorhynchus, have constantly increased. Thus, management of insecticide resistance to major insecticides including pyrethroids and organophosphates is required for more efficient control of resistant populations. In this study, the quantitative sequencing (QS) protocols were established to detect the frequencies of three mutations (the L1014F on voltage sensitive sodium channel and the G119S and F331W on acetylcholinesterase 1) that are associated with either pyrethroids or organophosphates. Based on the QS protocol using newly designed non-polymorphic primers, resistance allele frequencies (RAFs) were estimated in field populations of An. sinensis, Cx. pipiens and Cx tritaeniorhynchus collected from an identical site in Korea. The dynamics of each resistance allele frequency over time in the same populations were also evaluated.

Until recently, there have been many researches about the freezing methods and several methods of cryopreservation. Hypothermic preservation has been used to complement the embryo freezing technology. There is a study to show the successful results for long-term hypothermic preservation. For that reason, FBS and BSA are commonly added to the culture medium to support embryo development. We investigated the effectiveness of hypothermic preservation method at 4℃ according to embryonic developmental stages for Hanwoo embryos and evaluated the effect of FBS and BSA on Hanwoo embryos as a supplemental reagent in hypothermic preservation medium after recovering preserved embryos from hypothermic preservation. The present study reported that survival and hatching rates of embryos at morula stage following storage at 4℃ is Day 7 group was significantly higher (p < 0.05) compared than those of other groups (p < 0.05). As a result, the survival and hatching rates of embryos at the blastocyst stage following storage at 4℃ result is showed that significantly higher (p < 0.05) survival rates than those of other groups an Day 6. The result showed that hatching rate at Day 6 and 7 were significantly lower (p < 0.05) compared with other groups. The result regarding the survival and hatching rates of bovine embryos following storage at 4℃ for 72 h in various concentrations of BSA are shown The results showed that survival rate of 1% BSA group was not significantly different (p < 0.05) compare with control (FBS) group. Also, the results showed that hatching rate of control (FBS) and 1% BSA were significantly different (p < 0.05) compared with other groups. In conclusion, our result demonstrated that the hypothermic preservation did not effect on the survival and hatching rates of embryos after recovering. In addition, the supplementation of BSA in preservation medium showed no difference in the embryo developmental competence after hypothermic preservation compared to FBS treatment. With that, BSA can be an alternative reagent for the hypothermic preservation medium as an energy source and pH buffer.

The hypopharyngeal gland (HPG) of the honeybee worker produces royal jelly (RJ) and has a developmental cycle closely related to the division of labor. In this study, we investigated to compare the HPG acini diameter of differently aged worker bees with high royal jelly producing colony (HRC) or less producing colony (LRC). Additionally, we also evaluated whether the fresh weight of the head is a reliable indicator of the developmental status of HPG. The HRC showed a significantly higher RJ production about two-times as compared with those of the LRC. We measured the HG-diameters on days 1, 3, 6, 9, 12, 15. The microscopic analysis revealed that the acini size of the HRC was significantly larger than the LRC. In addition, the acini diameter of HRC was 15% longer than the LRC on the first day after emerging. It was shown that the fastest development during 3 days which is preparing for nurse the brood. The HPG acini diameters increased in both colonies in a similar fashion until day 12 and then decreased. We also compared the fresh head weight of the experimental colonies, differences were similar to the development of HPG. Therefore, high royal jelly production may have a positive correlation between HPG acini size and the fresh head weight.

벼메뚜기는 분류학적으로 메뚜기목(Orthoptera), 메뚜기과(Acrididae)에 속하는 곤충으로 작물에 피해를 주는 해충으로 인식하고 있지만 예로부터 단백질 보충을 위하여 채집하여 튀기거나 볶음요리 로 이용해 왔다. 벼메뚜기는 갈색거저리, 쌍별귀뚜라미 등과 함께 식품공전에 등록되어 있으나 1년 1세대로 가을철에 채집하여 이용하는 실정으로 공급확대에 한계가 있다. 최근 우리나라에서 벼메뚜기를 사육하고자 하는 농가와 다양한 용도로 가공을 통해 이용하려는 수요가 점차 증가하고 있으나 연중사육기술이 개발되지 않아 대량사육하는 농가가 거의 없는 실정이다. 이에 벼메뚜기의 연중대량사육기술 뿐만 아니라 인공사료 및 자동화 사육시설 개발에 따른 생산비 절감 기술 개발이 필요한 실정이다. 벼메뚜기의 먹이로는 여름작물로 옥수수, 수단그래스, 겨울작물로 밀, 보리 등의 볏과작물이 있으며 많은 양의 생엽과 발육에 적합한 작물의 선발 및 볏과작물의 통곡실 가루를 이용한 인공사료의 개발로 먹이공급 부족 시 15~20일 급여 가능하며, 벼메뚜기 사육시설로 단독형, 연결형, 절충형 등의 사육시설을 개발하여 연결형 사육시설의 수확량과 생존율이 가장 높았으며, 예취 급이 노동력이 단독형의 16% 소요되었다.

To identify the venom components and their expression patterns of some Aculeata bees/wasps, venom gland-specific transcriptome analysis was conducted. FPKM values were normalized with the average of the transcription level of reference gene (a-tubulin). Common components in both solitary and social wasp venoms include hyaluronidase, phospholipase A2, metalloendopeptidase, etc. Although it has been expected that more diverse bioactive components with the functions of prey inactivation and physiology manipulation are present in solitary wasps, the information on venom compositions of solitary wasps obtained in this study was not sufficient to generalizae this notion. Nevertheless, some neurotoxic peptides (e.g., pompilidotoxin and dendrotoxin-like peptide) and proteins (e.g., insuline-like peptide binding protein) appear to be specific to solitary wasp venom. In contrast, several proteins, such as venom allergen 5 protein, venom acid phosphatase, and various phospholipases, appear to be relatively more abundant in social wasp venom. In the venom gland trancsriptome of bumblebees, major allergens or pain producing factors were barely identified, implying that bumblebee venoms are relatively less toxic than those of social or solitary wasps.

Somatic cell nuclear transfer (SCNT) is the technique which generates embryos by transferring diploid nucleus into an enucleated oocyte, it has produced specific animals successfully in a variety of species. However, the developmental capacity of SCNT embryos is still relatively lower than that of embryos produced in vivo. Oocyte is a kind of lipid rich cells, its quality limits the efficiency of embryo production. L-carnitine is a co-enzyme facilitating the transportation of long chain fatty acids across the inner mitochondria membrane where fatty acids are used for generating adenosine triphosphate (ATP) via beta-oxidation. It also has antioxidant actions which may protect mitochondrial membranes and DNA against damage induced by reactive oxygen species (ROS). Whether L-carnitine is functional in bovine SCNT embryos are unknown. Therefore, the objective of this study was to examine the effects of L-carnitine on oocyte maturation and developmental competence of subsequent SCNT embryos. L-carnitine was supplemented during IVM, then intracellular ROS and GSH levels, mitochondrial activity, gene expression of COCs were analyzed at the end of IVM. SCNT embryos were produced subsequently, apoptosis detection and gene expression evaluation were performed in blastocysts. In the results, treatments with 1.5 mM and 3 mM L-carnitine significantly improved maturation rates (P<0.05). Treatments with 3 mM L-carnitine effectively induced improvement in nuclear maturation, intracellular GSH levels and mitochondrial activity, as well as a reduction in intracellular ROS levels (P<0.05). mRNA levels of CPT1A, ACAA1, ACAA2, AREG, EREG, SOD1, GPX4, GLUT1 and CDC2 transcripts were effectively up-regulated by 3 mM L-carnitine treatments (P < 0.05). Similarly, 3mM L-carnitine induced an increase in blastocyst developmental rates and an improvement in blastocyst quality (P<0.05). Our study indicates that L-carnitine treatment during IVM improves oocyte nuclear maturation and subsequent SCNT embryo development.

Somatic cell nuclear transfer (SCNT) has been considered for preserving genetically valuable or endangered animals. Sapsaree is a Natianal Monument in Korea to maintian a pure pedigree. The aim of this study was to produce azoospermia Sapsaree using SCNT and identify normal reproductive ability of cloned azoospermia Sapsaree. Ear skin biopsy was performed on a thirteen-year-old azoospermia Sapsaree and ear skin fibroblasts were isolated for SCNT as donor cells. The fibroblasts were injected into enucleated in vivo matured oocytes, the couplets were electrical fused by two pulse of direct current (55 V for 15 μs) using titanium and platinum fusion needle and activated by calcium ionophore. Cloned embryos were surgically transferred into oviducts of natuarally estrus cycle synchronized recipient dogs. The fusion rate of platinum needle was 70%, which was higher than those of titanium needle (64.1%). Developmental rate to the 8 cells and 10 cell stages was higher in platinum needle group (24% and 16%, respectively) than those of platinum needle group (14.8% and 3.1%, respectively). Total 35 SCNT embryos were transferred into oviducts of 3 recipient dogs and one recipient finally delivered a puppy by caesarean section. As results, this study demonstrated that platinum fusion needle could be successfully make the reconstructed embryos and improve the efficiency of canine SCNT. Cloning azoospermia Sapsaree may contribute to conserve genetically valuable and unique pedigree. And further study should be confirm whether cloned live dog is azoospermia.

Prolonged communication between oocytes and the surrounding somatic cells is one of the unique reproductive physiology in canine. Paracrine Kit ligand (KITL) signaling is a well-known communication between granulosa cells and the oocyte. KITL is a cytokine growth factor secreted by granulosa cells that signals via the c-kit receptor expressed by oocytes. Paracrine factors, including growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15), exert their effects by binding with the kinase receptors expressed on the granulosa cells. However, the regulations of GDF9 and BMP15 in the canine KITL expression are currently poorly understood. Therefore, we investigated the effects of GDF9 and BMP15 on the expression of KITL in canine ovarian granulosa cells in vitro. In Annexin V assay recombinant GDF9 and BMP15 did not induce apoptosis in the cultured ovarian granulosa cells. When treated, FSH significantly increased KITL expression, and hCG suppressed its expression. When both FSH and hCG were treated, the expression of KITL was affected by GDF9 and BMP15 in dose and time dependent manner in the luteal granulosa cells. GDF9 (10 ng/mL) significantly decreased KITL expression after12 h. BMP15 (10 ng/mL) significantly also decreased KITL expression after 24 h. Western blot and immunochemistry results indicate that GDF9 activated Smad2/3. After blocking ALK 4/5/7 receptors by SB, GDF9 failed to activate Smad2/3, also BMP15 did not activate Smad1/5/8 after blocking ALK 2/3/6 receptors by DM. So GDF9 exerts its effects via using ALK 4/5/7 receptors to activate SMAD2/3 signaling, and BMP15 binds ALK 2/3/6 receptors to activate SMAD1/5/8 signaling. The expression of KITL was not changed by SB or DM treatment. However, the effect of GDF9 and BMP15, which decreased the expression of KITL, was suppressed by SB or DM treatment. In conclusion, this study provides the first evidence that recombinant GDF9 and BMP15 decrease KITL expression in canine ovarian granulosa cells.

The cancer and Parkinson's disease associated protein DJ-1 is multifunctional protein that involves in diverse cellular process. DJ-1 protein has a cellular protective role and promoted cell survival under an oxidative stress. However, the cellular protective mechanism of DJ-1 is not fully understand, and we needs to be further study their functions in novel organisms. In the present study, we investigated the protective role of DJ-1 against induced oxidative stress in canine cell line. On the basis of these experiments, canine DJ-1 overexpressing and null cell lines were established. The stable overexpression and down regulation of DJ-1 efficiency confirmed by the western blot analysis. Subsequently, the DJ-1 gene transfected cell lines and control cells were subjected to induced the oxidative stress, and then cell viability, cell proliferation assay, cellular apoptosis detection analysis (Annexin V and TUNEL assay), intracellular ROS and mitochondrial activity were measured appropriately. The results showed that DJ-1 overexpressed cells were up-regulated cell viability under oxidative stress conditions induced by the rotenone and hydrogen peroxide (H2O2), whereas loss of DJ-1 cells were down-regulated the cell survival activity. Additionally, overexpression of DJ-1 cells increased cell resistance to oxidative stress and inhibited the elevation of cell death and cellular ROS induced apoptosis. Moreover, DJ-1 overexpressed cells was increased mitochondrial functions by using confocal microscopy with MitoTracker staining. On the contrary to this, DJ-1 null cells show defective cellular protection and mitochondria activity against oxidative stress conditions. Our data indicate that canine DJ-1 protein attenuates cellular apoptosis and ROS generation, enhances the cellular survival activity and promote mitochondrial function under the oxidative stress, likewise other mammalian cells. Importantly, DJ-1 overexpression may be an important part of a protective strategy as a sensor for oxidative stress.