We sequenced 17,329 bp of mitochondrial genome (mitogenome) of the black dwarf honey bee, Apis andreniformis (Hymenoptera: Apidae), that lacks ~200 bp of the A+T-rich region for the completion of the genomic sequence. The gene arrangement of A. andreniformis mitogenome is identical to that of A. cerana. However, the genome contains 5 additional tRNALeu(CUN) located 4 copies between tRNAMet and tRNAGln, and 1 copy between tRNAGln and tRNAAla, along with the typical sets of genes (13 protein-coding genes, 22 tRNAs, and 2 rRNAs) including regular tRNALeu(CUN) and the A+T-rich region (at least 923 bp). Only 1 copy of tRNALeu(CUN) differed by 1 bp from other 4 copies of tRNALeu(CUN). Each additional tRNALeu(CUN) is followed by nearly identical 68-bp long repeat sequence (95.6% identity). All 13 protein coding genes have typical start codons found in insect mitochondrial PCGs (2 ATA, 9 ATT, and 2 ATG).

Patients referred to the Veterinary Medical Center at Chungbuk National University were analyzed. This study aimed to provide information to clinicians in Korea on the factors affecting fracture incidence in patients adapted to Korean companion animal culture. Eighty cases showing- bone fractures were investigated from January, 2005 to De- cember, 2011. Fractures were mainly observed in miniature dogs such as Yorkshire terrier (12%), Poodle (12%), and Maltese (9%). Almost 50% of all cases were less than 1 year old. Total bone fractures in summer and fall constituted 62%. Fracture incidence was higher in male dogs (54%) than female dogs (46%). Hindlimb fracture (37%) was the most common. Distribution of limb fractures was higher in the femur (19), followed by the tibia/fibula (15), radius/ulna (13), humerus (11), and mandible (8). Pelvic fracture was observed in 19 cases. Simple fracture (84%) was the most common fracture type, followed by comminuted fracture (16%). Distal portion was the most common fracture site in long bones (67%), followed by middle (26%) and proxi- mal bones (7%). Fractures were mainly caused by traffic accidents (43%), whereas 28.5% of fractures were due to falling down. Plate & screw (26%) was the most common method of fracture repair, followed by intramedullary pin (24%), wire (11%), external skeletal fixation (10%), screw (9%), amputation (7%), cross pin (6%), IM pin and wire (5%), and rush pin (1%). Among 80 patients, 79%of pa- tients showed a good prognosis after surgery, whereas 13% patients did not experience a good result. This study shows that miniature dogs are prone to falling and fractures, espe- cially in the distal part of long bones.

An efficient plant regeneration protocol is developed for a standard-type chrysanthemum. When petal segments derived from flower buds (4 or 8cm in diameter) were used as the culture material, the highest shoot regeneration frequency (96%) was obtained on a Murashige and Skoog (MS) medium supplemented with 0.5 mg/L IAA, 2 mg/L BA, 3% sucrose, and a 0.8% agar. Pre-culturing the explants under dark conditions for 14 days produced better results for the shoot regeneration frequency than the explants cultured under a continuous 16 h photoperiod (40 μmolm-2 s -1 ). The shoot regeneration frequency ranged from 19.0% for the Shinmato cultivar to 89.1% for the Baeksun cultivar. Activated charcoal (0.2%) enhanced the root formation of the regenerated shoots in a hormone-free MS medium. The rooted plantlets were acclimatized and successfully established in a greenhouse.

Brucellosis is an important bacterial zoonosis in humans and domestic animals. Brucella spp. are taken up, and survive within non-professional and professional phagocytes. In common belief, diabetes mellitus increases susceptibility to pathogenic infection. In this study, Brucella (B.) abortus was inoculated into a diabetic animal model, db/db mice, in order to show the course of brucellosis in diabetic state. The liver proliferation, bacterial burden of the liver, level of cytokines in serum and macrophage migration into liver, were investigated at 14 days post-infection. In comparison with the uninfected control mice, the results revealed that the weight of the liver of infected db/db mice was higher but with lower bacterial load in this organ. The level of MCP-1 mRNA expression in the liver was lower, the levels of IL-12p70, IL-10, TNF-α and IFN-γ in serum was significantly higher and the macrophages migration was significantly lower in infected mice than in the control group. In conclusion, this present study suggested that MCP-1 suppression by B. abortus infection may inhibit the macrophages migration, and consequently may induce to abrogate the bacterial survival in db/db mouse liver. Furthermore, the increased inflammatory cytokines may contribute to inhibition of B. abortus proliferation in diabetic mice.

Hipparchia autonoe belongs to the family Nymphalidae (Lepidoptera) and is designated as an endangered insect and national monument in Korea. It only inhabits a very restricted area on Mt. Halla but is widely distributed in several Asian countries including Mongolia. A previous study conducted to understand the genetic relationship between Mt. Halla and Mongolian H. autonoe for conservation purposes suffered from a limited number of samples. Therefore, we sequenced the DNA barcode region of an additional 36 H. autonoe individuals, combined them with previous data from 19 individuals, and performed phylogenetic and population genetic analyses. Furthermore, the internal transcribed spacer 2 (ITS2) region was also sequenced from the 36 samples as a nuclear DNA marker. The existence of independent haplotypes, sequence types, and significant FST estimates (p < 0.05) between Mt. Halla and Mongolian populations indicated hampered gene flow between the populations. Nevertheless, an absence of a reciprocal monophyletic group in Mt. Halla and Mongolian populations by cytochrome oxidase subunit I gene- and ITS2-based phylogeny suggests that the genetic isolation of the Mt. Halla population from the Mongolian populations seemed not large enough to consider them independent genetic entities.

In the present study, the 17,694-bp long complete mitochondrial genome (mitogenome) of the dwarf honey bee, Apis florea (Hymenoptera: Apidae), is described with an emphasis on the noteworthy triplicated tRNAser(AGN) region and an extraordinary long A+T-rich region with repeat regions. The gene arrangement of A. florea mitogenome is identical to that of A. mellifera, but has triplicated tRNASer(AGN), each of which contains the precedent 44 bp-long and following another 64 bp-long repeats plus one complete first repeat abutting to tRNAMet. A total of 1,610-bp long two repeat regions in 1,987 bp-long A+T-rich region is composed of nearly identical 141 ~ 219-bp long five tandem repeats and 50 ~ 52-bp long 12 tandem repeats that are encompassed by three non-repeat sequences. One of the common interpretations for such repeat sequence is slipped-strand mispairing and unequal crossing-over events during DNA replication.

The deleted in colorectal cancer (DCC) protein mediates attractant responses to netrin during axonogenesis. In the rat trigeminal ganglia (TG), axons must extend toward and grow into the trigeminal nerve to innervate target tissues such as dental pulp. Our present study aimed to investigate the exp- ression of DCC in the TG. Four developmental timepoints were assessed in the experiments: postnatal days 0, 7 and 10 and adulthood. RT-PCR and western blotting revealed that the expression of DCC mRNA and protein does not signi- ficantly change throughout development. Immunohistoche- mistry demonstrated that DCC expression in the TG was detectable in the perikarya region of the ganglion cells du- ring development. Nerve injury at 3 and 5 days after the man- dibular nerve had been cut did not induce altered expression of DCC mRNA in the TG. Moreover, DCC-positive cell bodies also showed similar immunoreactive patterns after a nerve cut injury. The results of this study suggest that DCC constituti- vely participates in an axonogenesis attractant in ways other than expression regulation.

As the increase of chrysanthemum demand on chrysanthemum increases in Korea, the production of high quality chrysanthemum is needed. Chrysanthemum stunt viroid (CSVd) is one of the important viroid, which infects chrysanthemum and induces diseases that affects the decrease of quality and yield. To solve this problem, we used different size of meristem of chrysanthemum ‘Ency’ for shoot tip culture and also that of combined with heat treatment at 37οC. The efficiency of CSVd elimination was influenced by the size of shoot tip. The small-sized of meristems with 1 or 2 leaf primodia were regenerated into the highest number of CSVd-free plantlets. By RT-PCR, the 214-bp band corresponding to CSVd was not detected in 22.2% of the total number of tested regenerants from shoot tips with 2 leaf primordia. While, shoot tip culture combined with heat treatment of one-month-old in vitro shoots was not effective for CSVd-elimination. The CSVd-free plants grew more vigorously than CSVd-infected plants in the greenhouse.

Salmonellosis is a widespread bacterial zoonosis that commonly causes enterocolitis and foodborne poisoning leading to an extensive economic loss in domestic animal industry. Considerably, the emergence of multidrug resistant strains of Salmonella spp. induces further severe problems affecting public health. The present report was designated to investigate the antibacterial efficacies of three common disinfectants including an oxidizing compound disinfectant (OXC), a triple salt (TS) and a quaternary ammonium compound (QAC) against Salmonella typhimurium subjected to the preliminary changes of drug temperature. All solutions of three disinfectants were pre-incubated at different temperature (22, 37 and 63°C) for 1 h prior to exposure to bacteria. The disinfectants and bacteria were diluted with distilled water (DW), hard water (HW) or organic matter suspension (OMS) according to treatment condition. Under the DW condition, the disinfectant efficacy of the QAC at 63°C was higher than that of 22°C. Furthermore, under HW diluent the disinfectant efficacy of the TS pre-warmed at both of 37 and 63°C were increased compared to that of 22°C. Considerably, the efficacy of pre-warmed QAC at both of 37 and 63°C under the OMS diluent were higher than that of 22°C. Conclusively, prewarming at higher temperatures have positive effects on the stability of the antibacterial efficacies of TS and QAC.

In this study, we examined whether Hanganutziu‐Deicher (H‐D) antigens are important as an immunogenic non‐a1,3‐galactose (Gal) epitope in pigs with a disrupted a1,3‐ galactosyltransferase gene. The targeting efficiency of the AO blood genotype was achieved (2.2%) in pig fibroblast cells. A total of 1800 somatic cell nuclear transfer (SCNT) embryos were transferred to 10 recipients. One recipient developed to term and naturally delivered two piglets. The a1,3‐galactosyltransferase activity in lung, liver, spleen, and testis of heterozygote a1,3‐galactosyltransferase gene knockout (GalT‐KO) pigs was significantly decreased, whereas brain and heart showed very low decreasing levels of a1,3‐ galactosyltransferase activity when compared to those of control. Enzyme‐linked lectinosorbent assay showed that the heterozygote GalT‐KO pig had more sialyla2,6‐ and sialyla2,3‐ linked glycan than the control. Furthermore, the heart, liver, and kidney of the heterozygote GalT‐KO pig had a higher N‐glycolylneuraminic acid (Neu5Gc) content than the control, whereas the lung of the heterozygote GalT‐KO pig had Neu5Gc content similar to the control. Collectively, the data strongly indicated that Neu5Gc is a more critical xenoantigen to overcoming the next acute immune rejection in pig to human xenotransplantation.

Genomic reprogramming factors in the GV cytoplasm improved cloning efficiency in mice through the pre‐exposure of somatic cell nuclei to a GV cytoplasmic extract prior to nuclear transfer. In this study, a pig GV oocyte extract (pGV extract) was developed. Treatment of pig fibroblasts with the pGV extract promoted colony formation after 2–3 weeks in culture, concomitant with the expression of stem cell markers (Oct‐4, Rex1, Nanog, Sox2) and repression of differentiated cell markers (CKAP2, NPR3 ). Using fibroblasts transfected with human Oct‐4 promoter‐driven enhanced green fluorescent protein (Oct4‐EGFP), pGV extract treatment induced the reactivation of the Oct‐4 promoter in Oct4 ‐ EGFP cells by 10 days post‐treatment. These transgenic donor cells were injected into 8‐cell embryos. Oct‐4 promoter activity was subsequently detected in most ICM cells of the host blastocyst. Interestingly, reconstructed embryos with pGV extract‐treated Oct4‐ EGFP fibroblast nuclei showed prolonged expression of Oct4 in the ICM of embryos. Additionally, the pGV extract promoted somatic cell reprogramming and cloned embryo development when assessed by measuring histone H3‐K9 hypomethylation, the expression of Oct4 and Nanog in blastocysts, and the production of increased numbers of high‐ quality blastocysts. Under specific culture conditions, pGV extract‐treated fibroblast cells differentiated into neuronal, pancreas, cardiac, and endothelial lineages that were confirmed by antibodies against specific marker proteins. These data provide evidence for the generation of stem‐like cells from differentiated somatic cells by treatment with GV oocyte extracts in pig.

Accumulating evidence suggests that chemotherapy can cause long‐term detrimental effects and alter the biology of the recipient environment. Therefore, a subsequent report claimed that the transplantation of female germline stem cells (FGSCs) into the ovaries of recipient mice that were pretreated with a high dose of busulfan and cyclophosphamide (B/C) resulted in the successful production of offspring. Therefore, this study was conducted to further clarify the impact of female germ cell transplantation on female ovaries after B/C treatment. RT‐PCR analysis showed that the period of germ cell depletion coincide with decreased Figla, Lhx8, Nobox, Kit, and Sox3 gene expression in the B/Ctreated ovary. However, depletion of female germ cells is mediated by a Fas/FasL‐, Kit/ Kitl‐, TNF‐, p53‐ and autophagy‐ independent pathway. Also, histological analysis is similar to that of Nobox null‐derived ovaries, indicating that follicle death after B/C treatment might be caused by down‐regulating of Nobox pathway. When female mice during 15 weeks after B/C treatment were checked for reproductive activity, B/C treated mice did not produce their pups. In addition, when 3×106 GFP positive primordial follicles were injected into B/C treated female mouse ovaries, donor follicle were not able to colonize into the ovaries of recipients. In conclusion, these data from a preclinical mouse model strongly suggested that female ovary until 15 weeks after B/C treatment could not support environment for maturing of exogenous FGSCs.

The objective of this study was to assess the effect on post-thawed sperm motility, viability and acrosome integrity of boar semen frozen in the freezing extender with chicken or duck egg yolks. The Sperm rich fraction of ejaculates from three Duroc boars were collected by a glove-hand technique. Samples with more than 80% motile sperm were used for this experiment. Semen was diluted with freezing extender (LEY) containing 11% (v/v) lactose, 20% (v/v) hen egg yolk with 3.5% (v/v) glycerol, and 0.5% (v/v) Orvus Es Paste(OEP, Nova Chemical Sales Inc., Scituate, MA. USA) to yield a final sperm concentration of 5×108 cells/ml. Following complete dilution, semen samples were loaded in 0.5 ml French medium straws (IMV technologies, France) and transferred to programmable semen freezer (SY-LAB Gerate GmbH, Austria). For freezing the semen samples, each straw was cooled from 5℃ to — 5℃ at 6℃/min, auto-seeding at — 5℃ and held for 60sec, samples were then cooled from — 5 to — 80℃ at 40℃/min, and thereafter from — 80℃ to — 150℃ at 60℃/min. The yolks used were sourced from fresh chicken and duck eggs. To evaluate the post-thaw sperm quality, semen was thawed at 38℃ for 20 sec and sperm motility, viability and acrosome integrity were assessed. Motility was assessed for %motile cell characteristics using computer-assisted semen analysis (CASA; SAIS SI-100, Medical supply, Korea). The percentage of sperm viability was assessed using LIVE/DEAD® sperm viability kit (Molecular probes, Eugene, OR, USA). The acrosome integrity was assessed by FITC-PNA staining. Sperm quality in terms of motility, viability and acrosome integrity showed higher after freezing in medium containing duck yolk than chicken yolk. However, there was no significant difference in sperm quality for the different types of yolk(p>0.05). * The result of this study showed that there was no significant difference between the egg yolk types when considering the sperm motility, viability and acrosome integrity of boar semen frozen in the freezing extender with chicken or duck egg yolks.

There are diverse methods of cryopreservation of mammalian embryos with variable degrees of success. Although cryopreservation technique of mammalian embryos has been advanced, freezing stress affect to cellular event such as apoptosis and autophage in embryos. The objective of the study is to investigate the affection of to survival, development, live offspring, apoptosis and autophagy on embryo. Mouse embryos were vitrified and thawed using normal straw and modified cut standard straw (M-CSS), then in vitro cultured until blastocyst stage and transferred to recipient. Recovery rates (100 vs 99.2%), survival rates (99.2 vs 78.6%), developmental rates (18.4 vs 10.7%), total cell numbers (45 vs 37), preganacy rates (34.5 vs 25%) and offspring numbers (10.1 vs 4.9 %) of M-CSS group are significantly higher than those of normal straw vitrified group. Also, rate of apoptosis in blastocysts developed using M-CSS (1.9%) was significantly lower than using normal straw vitrification (2.7%). Apoptosis-related gene, caspase 3, was expressed at the highest level in blastocysts derived from normal straw group. However, no differences of autophagy related gene, Atg6 and expression of LC3 between normal straw and M-CSS groups were observed. In conclusion, the standard vitrification procedure induces mitochondrial apoptosis in zygotes in an autophagy-independent manner, whereas the novel M-CSS procedure may improve embryo vitrification.