During January-November 2012, a total of 2,041 quarter milk samples were collected from dairy cattle of 82 dairy farms nationwide. About 42% (870/2,041) of the samples that had somatic cell counts (SCC) of ≥ 200,000 cells/ml were subjected to microbiological examination. No bacteria was isolated from 95 of 870 (10.9%) samples. Among 1,237 bacteria isolated from the rest 775 samples, 1,085 were identified with VITEK: more than half (52.1%, 645/1,237) of the isolates were gram negative bacillus. Gram positive cocci including Staphylococcus accounted for 35% of the isolates and almost none of gram positive bacilli isolated. Excluding Coagulase-negative Staphylococcus (CNS), the most frequently isolated bacterial species was Escherichia coli (11.2%, 138/1, 237), followed by Klebsiella pneumoniae (8.1%,100/1,237), Staphylococcus aureus (7.1%, 88/1, 237), Enterobacter cloacae (6.0%, 74/1, 237), and Serratia marcescens (3.5%, 43/1, 237). The most common resistance of S. aureus was to penicillin (77.4%) and ampicillin (73.0%), while no resistance was observed against gentamicin and cephalothin. Although CNS presented resistance to all antimicrobials tested but the most prevalent resistance was to penicillin (35.6%) and ampicillin (37.0%). The pattern of antimicrobial resistance observed in CNS was similar to that of S. aureus, but the rates were much lower than those of S. aureus. E. coli also showed resistance to all the antimicrobials tested, although the rates were not very high. The highest resistance of E. coli was to cephalothin (39.4%) and ampicillin (36.2%), while most of the strains (98.0%) showed sensitivity to amikacin. The results of this study provide information on current situation of bovine mastitis in Korea.

This paper describes the epidemiological characteristics of bovine tuberculosis in Korea during January 2000 to September 2004, when the incidence of bovine tuberculosis increased markedly: a total of 1,054 herds (4,197 cattle) were confirmed to be infected with Mycobacterium bovis during this period. Based on the record of epidemiological investigation, introduction of purchased cattle (22.9%, 125/545) into a farm was the most frequent transmission route of M. bovis infection. On 31.7% (335/1,054) of the infected farms, recurrent infection occurred more than once before the disease has been eradicated completely. The highest rate of recurrence was detected around 70 days after the initial test of the infected herd, which seems to be related to current regulation on the test of animals that cohabited with those previously diagnosed with infection in farms, rather than to the characteristic of the disease. Although the current eradication program has been effective in controlling the disease in dairy cattle in Korea, control measures more specific to beef cattle may be needed because infection rate in beef cattle continues to increase in recent years.

To determine current rate of antimicrobial resistance, a total of 236 isolates from milk samples of dairy cattle with mastitis in Korea during 2010-2011 were examined against 12 antimicrobials using disc diffusion method: 67 Staphylococcus aureus, 74 coagulase-negative Staphylococcus spp. (CNS), and 95 Escherichia coli isolates. The isolates examined in this study were submitted by Local Veterinary Service Laboratories located in 13 provinces and metropolitan cities nationwide. The highest rates of resistance among S. aureus isolates were against ampicillin (56.7%) and penicillin (56.7%), followed by kanamycin (11.9%). All S. aureus isolates were sensitive to lincomycin, amikacin, and cephalothin. Only one isolate showed resistance to tetracycline and oxacillin, respectively. Less than 10% of the S. aureus isolates presented resistance to erythromycin, neomycin, and gentamicin. Among CNS isolates, the most frequently observed resistance was to lincomycin (44.5%), followed by penicillin (28.3%), ampicillin (18.9%), tetracycline (17.5%), kanamycin (13.5%), and erythromycin (9.4%). All or most of the CNS isolates were sensitive to cephalothin, amikacin, neomycin, and gentamicin. The highest rate of resistance among E. coli isolates was against tetracycline (26.3%), followed by streptomycin (21%), neomycin (15%), kanamycin (12.6%), and gentamicin (10.5%). Amikacin was the only antimicrobial to which no E. coli isolates showed resistance. Around 10% of the S. aureus isolates and 15% of the CNS isolates showed resistance against three or more antimicrobials simultaneously, while more than 30% of the E. coli isolates did.

The ovarian development and oviposition response of two vector beetles of pine wood nematode, Monochamus alternatus and M. saltuarius, was investigated. The ovary structure of two beetles was investigated by means of light microscopy. The ovary of both beetles was composed of two ovaries that are connected by a common oviduct. Each of the two ovaries was consisted of twelve ovarioles, the functional units of female oogenesis. The ovary type was meroistic telotrophic with nurse cells and oocytes in the tropharium. Ovarian development of two vector beetles was completed at 12-14 days after maturation-feeding pine twigs. Aspect of accumulation of vitellogenins in the ovary of two vectors showed difference in developmental stages and major yolk proteins, differently from vitellogenin. To investigate oviposition performance of two vector beetles, we provided P. thunbergii and P. koraiensis bolts for egg laying and collected emerged adults from each pine bolts in the next year. M. saltuarius females made more oviposition wounds and entrance hole of larvae than M. alternatus on pine bolts. We also investigated whether two beetles can transmission of Bursaphelenchus xylophilus and the European type of B. mucronatus via oviposition to noninfected dead pines. 48-83% from newly emerged adults of two beetles were carried two species nematode. This results suggested that two species beetles can transmit nematodes through oviposition performance and transmitted nematodes successfully propagate in non-infected dead pines.

A survey was conducted in the northern conifer forests of Korea from February to May in 2009 for the Monochamus species vectored Bursaphelenchus species. Dead pine trees infested with bark beetle larvaes were collected from Pinus densiflora and P. koraiensis located in the following target areas: Pocheon, Yeongi, Jincheon, Chuncheon, and Bonghwa. A cerambycid beetle, M. saltuarius was only found from sampled log piles in target areas. Bursaphelenchus species carried by M. saltuarius was only isolated in Pocheon, Gyeonggi province. The nematodes recovered from 17 (71%) of 24 beetles from P. koraiensis and 12 (75%) of 16 beetles from P. densiflora. The average number of nematode per adult was 1,974 and 11,823 from P. koraiensis and P. densiflora, respectively. The nematode was also recovered from the inner bark of both Pinus species infested with M. saltuarius larvae. Bursaphelenchus species identification was performed by morphological characters and molecular analysis with ITS-RFLP patterns and sequences of ITS and D2D3 region. Both morphological and molecular characters match well with the original description of the European type of B. mucronatus. This is the first report of M. saltuarius species as a vector of the European type of B. mucronatus on both P. densiflora and P. koraiensis in Korea.

Since pine wilt disease was first reported in Busan in 1988, this disease has become a serious threat to pine forests in Korea. A general survey for Bursaphelenchus xylophilus group (B. xylophilus, B. mucronatus) nematodes were carried out to determine their geographic range, host tree species and insect vectors, Monochamus species. Based on the survey results, B. xylophilus were found over 57 cities or district and a similar nematode, B. mucronatus also has been isolated over 30 cities or district including Jeju and Pocheon. The Asian type of B. mucronatus was founded from Pinus thunbergii and P. densiflora, generally associated with M. alternatus as its insect vector. The European type of B. mucronatus which was limited distribution to the central areas, such as Yeongwol, Bonghwa, Yeongdeok, was also found on P. koraiensis and P. desiflora associated with M. saltuarius. Both Monochamus species transmitted both Bursaphelenchus species and co-existed in Muju and Yeongdeok predicted as their range margin. Their range margins shifted northwards compared to 2006, seemingly as a result of climate change. Recently, the European type of B. mucronatus has been identified in other several areas since it was first reported in Pocheon in 2008. Several reports indicate the potential pathogenicity of European type of B. mucronatus and other species, B. sexdentati and B. lenoni in Europe. Therefore, sustainable survey and the study for the newly invading Bursaphelenchus species as well as the member of B. xylophilus group should be performed to protect Korean forests.

Monodispersed and nano-sized Cu powders were synthesized from copper sulfate pentahydrate inside a nonionic polymer matrix by using wet chemical reduction process. The sucrose was used as a nonionic polymer network source. The influences of a nonionic polymer matrix on the particle size of the prepared Cu powders were characterized by means of X-ray diffraction), scanning electron microscopy), and particle size analysis). The smallen Cu powders with size of approximately 100 nm was obtained with adding of 0.04M sucrose at reaction temperature of . The particle size of the Cu powders prepared by the reduction inside polymer network was strongly dependent of the sucrose content and reaction temperature.

Rice transformation method using A. tumefaciens has already been widely used to generate transgenic plants, the transformation rate is still low in most Korean elite cultivars. We made several modifications of the standard protocol especially in the co-cultivation step to improve the efficiency of the rice transformation. The co-culture medium was modified by the addition of three antioxidant compounds (10.5㎎/ℓ L-cysteine, 1mM sodium thiosulfate, 1mM dithiothreitol) and of Agrobacterium growth-inhibiting agent (5㎎/ℓ silver nitrate). Co-cultivation temperature (23. 5℃ for 1 day, 26.5℃ for 6 days) and duration (7 days) were also changed. The plasmid of pMJC-GB-GUS carrying the GUS reporter gene and the bar gene as the selectable marker was used to evaluate the efficiency of the transformation. After co-cultivation, a high level of GUS gene expression was observed in calli treated with the modified method. It is likely that those newly added compounds helped to minimize the damage due to oxidative bursts during plant cell-Agrobacterium interaction and to prevent necrosis of rice cells. And the transformation rate under the modified method was also remarkably increased approximately 8-fold in Heungnambyeo and 2-fold in Ilmibyeo as compared to the corresponding standard method. Furthermore, we could produce the transgenic plants stably from Ilpumbyeo which is a high-quality rice but its transformation rate is extremely low. Transformation and the copy number of transgenes were confirmed by PCR, bar strip and Southern blot analysis. The improved method would attribute reducing the effort and the time required to produce a large number of transgenic rice plants.

Rice transformation method using A. tumefaciens has already been widely used to generate transgenic plants, the transformation rate is still low in most Korean elite cultivars. We made several modifications of the standard protocol especially in the co-cultivation step to improve the efficiency of the rice transformation. The co-culture medium was modified by the addition of three antioxidant compounds (10.5㎎/ℓ L-cysteine, 1mM sodium thiosulfate, 1mM dithiothreitol) and of Agrobacterium growth-inhibiting agent (5㎎/ℓ silver nitrate). Co-cultivation temperature (23. 5℃ for 1 day, 26.5℃ for 6 days) and duration (7 days) were also changed. The plasmid of pMJC-GB-GUS carrying the GUS reporter gene and the bar gene as the selectable marker was used to evaluate the efficiency of the transformation. After co-cultivation, a high level of GUS gene expression was observed in calli treated with the modified method. It is likely that those newly added compounds helped to minimize the damage due to oxidative bursts during plant cell-Agrobacterium interaction and to prevent necrosis of rice cells. And the transformation rate under the modified method was also remarkably increased approximately 8-fold in Heungnambyeo and 2-fold in Ilmibyeo as compared to the corresponding standard method. Furthermore, we could produce the transgenic plants stably from Ilpumbyeo which is a high-quality rice but its transformation rate is extremely low. Transformation and the copy number of transgenes were confirmed by PCR, bar strip and Southern blot analysis. The improved method would attribute reducing the effort and the time required to produce a large number of transgenic rice plants.

Lysine is the first essential amino acid for optimal nutrient quality in rice grain. For the narrow genetic diversities of lysine contents in rice, somaclonal variation was the source of mutation in our breeding program. Biochemical selection was conducted using 1 mM S-(2-aminoethyl) cysteine followed by two passages of 5 mM lysine plus threonine in the callus subculture medium. The lysine contents in endosperm of all progenies recovered from the biochemical selection were higher than those of their donor cultivar 'Hwayeongbyeo'. These elevated lysine levels of mutants were successfully transmitted to M4 generation. The lysine contents in endosperm varied 3.85 to 4.80% compare to their donor cultivar 'Hwayeongbyeo' was 3.85%. Three of high-lysine germplasms, Lys-l, Lys-2 and Lys-7 were selected by biochemical selection and rapid screening methods. DNA analysis showed that a new insertion of Tos 17 which mapped to rice chromosome 11 on the high-lysine mutant, Lys-2.