Proteomic analysis of Salmonella pullorum infected liver tissue in chicken

Urantulkhuur.B,Baatartsogt.O,H. K. Lim,S.W Suh,Y.Y Kim,C.H Lee,P.K. Mandal,K. D. Choil 한국가금학회 2008 한국가금학회 정기총회 및 학술발표회 Vol.25 No.-

Salmonellosis in poultry is a significant health problem which causes substantial economic loss. One of the common causative agents of chicken salmonellosis is Salmonella pullorum (SP). The goal of this work was to characterize and identify gene expression profiles of chicken liver tissue after infected Salmonella pullorum. In particular, this report focuses on identification of the major proteins that could be identified in Salmonella infected liver. Hundred Hy-Line layer chicks of 10 days old were divided into 5 groups of 20 each. A1, A2, A3 group were infected with high dose(3×10⁹cfu/㎖) of S.P orally and A4, A5 group were kept as non-infected control. One chick was taken randomly from each group after 0, 3, 6, 9, and 12 days and liver was collected. Proteins were separated from 6 days chicks' liver. Two-dimensional electrophoresis was done and mass spectrometry(MS) will identify the protein spots. More than 289 protein spots were detected on silver stained 2DE gels. Three new proteins were found to be expressed in the infected chicken liver. Five proteins were found to have up regulated expression in infected chicken.

Isolation and characterization of a new Methanoculleus bourgensis strain KOR-2 from the rumen of Holstein steers

Urantulkhuur Battumur,이만희,배귀석,김창현 아세아·태평양축산학회 2019 Animal Bioscience Vol.32 No.2

Objective: To isolate and identify new methanogens from the rumen of Holstein steers in Korea. Methods: Representative rumen contents were obtained from three ruminally cannulated Holstein steers (793±8 kg). Pre-reduced media were used for the growth and isolation of methanogens. Optimum growth temperature, pH, and sodium chloride (NaCl) concentration as well as substrate utilization and antibiotic tolerance were investigated to determine the physiological characteristics of the isolated strain. Furthermore, the isolate was microscopically studied for its morphology. Polymerase chain reaction of 16S rRNA and mcrA gene-based amplicons was used for identification. Results: One strain designated as KOR-2 was isolated and found to be a non-motile irregular coccus with a diameter of 0.2 to 0.5 μm. KOR-2 utilized H2+CO2 and formate but was unable to metabolize acetate, methanol, trimethylamine, 2-propanol, and isobutanol for growth and methane production. The optimum temperature and pH for the growth of KOR-2 were 38°C and 6.8 to 7.0, respectively, while the optimum NaCl concentration essential for KOR-2 growth was 1.0% (w/v). KOR-2 tolerated ampicillin, penicillin G, kanamycin, spectromycin, and tetracycline. In contrast, the cell growth was inhibited by chloramphenicol. Phylogenetic analysis of 16S rRNA and mcrA genes revealed the relatedness between KOR-2 and Methanoculleus bourgensis. Conclusion: Based on the physiological and phylogenetic characteristics, KOR-2 was thought to be a new strain within the genus Methanoculleus and named Methanoculleus bourgensis KOR-2.

Isolation and characterization of new Methanosarcina mazei strains KOR-3, -4, -5, and -6 from an anaerobic digester using pig slurry

Urantulkhuur Battumur,윤영만,배귀석,김창현 아세아·태평양축산학회 2017 Animal Bioscience Vol.30 No.8

Objective: An experiment was conducted to isolate and identify new methanogens in Korea from an anaerobic digester that uses pig slurry. Methods: An anaerobic digestate sample was collected from an anaerobic digester using pig slurry. Pre-reduced media were used for the growth and isolation of methanogens. Growth temperature range, pH range, NaCl concentration range, substrate utilization, and antibiotic tolerance were investigated to determine the physiological characteristics of isolated methanogens. The isolates were also examined microscopically for their morphology and Gram-stained. Polymerase chain reaction of 16S rRNA and mcrA gene-based amplicons was used for identification purpose. Results: Four strains, designated KOR-3, -4, -5, and -6, were isolated and were non-motile, irregular coccoid, and 0.5 to 1.5 μm in diameter. Moreover, the cell walls of isolated strains were Gram-negative. KOR-3 and KOR-4 strains used acetate for methane production but did not use H2+CO2, formate, or methanol as a growth substrate KOR-5 and KOR-6 strains utilized acetate, methanol, and trimethylamine for methanogenesis but did not use H2+CO2 or formate as a growth substrate. The optimum temperature and pH for growth of four strains were 39°C and 6.8 to 7.2, respectively. The optimum concentration of NaCl for growth of KOR-3, KOR-5, and KOR-6 were 1.0% (w/v). The optimum NaCl concentration for KOR-4 was 0.5% (w/v). All of the strains tolerated ampicillin, penicillin G, kanamycin, streptomycin, and tetracycline; however, chloramphenicol inhibited cell growth. Phylogenetic analysis of 16S rRNA and mcrA genes demonstrated that strains KOR-3, -4, -5, and -6 are related to Methanosarcina mazei (M. mazei, 99% sequence similarity). Conclusion: On the basis of physiological and phylogenetic characteristics, strains KOR-3, -4, -5, and -6 are proposed to be new strains within the genus Methanosarcina, named M. mazei KOR-3, -4, -5, and -6.

메탄생성균의 순수배양시 메탄생성억제 화학물질의 첨가가 메탄생성에 미치는 영향

Urantulkhuur Battumur,이만희,김창현 강원대학교 동물생명과학연구소(구 강원대학교 동물자원공동연구소) 2017 동물자원연구 Vol.28 No.3

The objectives of the present study were to investigate the effect of five chemical inhibitors on methane production of the pure cultures of methanogens. To examine the methanogenesis inhibition by 2-bromoethane sulfonic acid (BES), 3-bromopropane sulfonic acid (BPS), lumazine, propynoic acid and ethyl 2-butynoate, they were added into the pure cultures of Methanobacterium formicicum KOR-1 and KOR-12, Methanoculleus bourgensis KOR-2, Methanosarcina mazei KOR-3, KOR-7, KOR-9 and KOR-10, Methanosarcina vacuolata (DSM 1232), Methanobacterium bryantii (ATCC 33272), Methanosarcina mazei (ATCC BAA159) and Methanosarcina barkeri (ATCC 43240). Each methanogen was anaerobically incubated at 39℃ for 6 and 10 days, and then methane gas production was measured. All methanogens were sensitive to BES, lumazaine and ethyl 2-butynoate and as the addition levels of the chemicals increased, the methanogenesis was linearly reduced. BPS and propynoic acid failed to inhibit any of the methanogens. There were important differences among species of methanogens regarding their sensitivity to the different inhibitors. In general, the strains of Methanosarcina species were the most resistant to BES, lumazaine and ethyl 2-butynoate and the strains of Methanobacteriales order the least resistant. The different resistances to chemical inhibitors may be caused by different cell membrane structure, substrate utilization for methanogenesis and competition with other microorganisms for electron receptors. In conclusion, differences among methanogens habituating in anaerobic ecosystems, such as the rumen of ruminants regarding their resistance to chemical inhibitors should be considered when strategies of inhibition of methanogenesis are designed.

Isolation and Characterization of a New Methanobacterium formicicum KOR-1 from an Anaerobic Digester Using Pig Slurry

Battumur, Urantulkhuur,Yoon, Young-Man,Kim, Chang-Hyun Asian Australasian Association of Animal Productio 2016 Animal Bioscience Vol.29 No.4

A new methanogen was isolated from an anaerobic digester using pig slurry in South Korea. Only one strain, designated KOR-1, was characterized in detail. Cells of KOR-1 were straight or crooked rods, non-motile, 5 to $15{\mu}m$ long and $0.7{\mu}m$ wide. They stained Gram-positive and produced methane from $H_2+CO_2$ and formate. Strain KOR-1 grew optimally at $38^{\circ}C$. The optimum pH for growth was 7.0. The strain grew at 0.5% to 3.0% NaCl, with optimum growth at 2.5% NaCl. The G+C content of genomic DNA of strain KOR-1 was 41 mol%. The strain tolerated ampicillin, penicillin G, kanamycin and streptomycin but tetracycline inhibited cell growth. A large fragment of the 16S rRNA gene (~1,350 bp) was obtained from the isolate and sequenced. Comparison of 16S rRNA genes revealed that strain KOR-1 is related to Methanobacterium formicicum (98%, sequence similarity), Methanobacterium bryantii (95%) and Methanobacterium ivanovii (93%). Phylogenetic analysis of the deduced mcrA gene sequences confirmed the closest relative as based on mcrA gene sequence analysis was Methanobacterium formicicum strain (97% nucleic acid sequence identity). On the basis of physiological and phylogenetic characteristics, strain KOR-1 is proposed as a new strain within the genus Methanobacterium, Methanobacterium formicicum KOR-1.

닭의 추백리 감염 후 간장 내의 단백체 변화 연구

김윤연,Urantulkhuur'B,Baatartsogt'O,서상원,김영교,안광철,최강덕 한국가금학회 2009 한국가금학회 정기총회 및 학술발표회 Vol.26 No.-

Salmonella pullorum in poultry is a signficant health problem which causes substantial economic loss. This study was conducted to characterize and identify protein expression profiles of chicken liver after S. pullorum challenge. To identify the differentially expressed chicken liver proteins after S. pullorum, we have used two-dimensional electrophoresis with MALDI-TOF mass spectrometry. Total 14 protein spots were selected from two-dimensional electrophoresis. 8 proteins were up-regulated and 6 proteins were down-regulated at the chicken liver infected S. pullorum. In conclusion, this study could be used to the determine characterize and identify protein expression profiles of chicken liver infected S. pullorum.

Immuno-regulatory Effect of Dryopteriscrassirhizoma Extract on Chicken Splenocyte

Baatartsogt. O,Urantulkhuur. B,S.W. Suh,H.K. Lim,Y.K. Kim,K.O.Park,J.H. Kim,S.H. Kim,P. K. mandal,K. D. Choi 한국가금학회 2008 한국가금학회 정기총회 및 학술발표회 Vol.25 No.-

We investigated the effect of herbal extract of Dryopteris crassirhizoma on the cell viability, immune response, and nitric oxide production of chicken splenocyte. Nitrite levels were assayed in the culture supernatant of splenocyte after 24h by the Griess method. The expressions of pro-inflammatory cytokines such as interleukin (IL-2, IL-6, IL-8, IL-18) and the housekeeping gene GAPDH were studied using RT-PCR. Splenocyte were cultured with the herbal extracts at 37℃ in 96-well plate for 24h. Not cytotoxic effect was observed at 20 ㎍ and 40 ㎍. The NO production increased dose dependently with the treatment of herbal extract. which suggest that this herbal extract has no anti-inflammatory effect. In cytokine expression it was observed that the herbal extract induced the expression of IL-2 and suppressed the expression of IL-6 and IL-18, however there was no effect on the expression of IL-8. It may be concluded that the Dryopteris crassirhizoma extract is non toxic and have no anti-inflammatory effect and it has immune stimulation effect through IL-2.

Bioremediation strategy for petroleum-contaminated soil in Mongolia

Buyan Chuluun,Ninjbadgar Batkhuyag,Urantulkhuur Battumur,Jeong Seong Pil 대한환경공학회 2019 대한환경공학회 학술발표논문집 Vol.2019 No.-

Detection and analysis of immunological genes in response to the Infection of Salmonella gallinarum in chicken

Baatartsogt. O,H. K. Lim,Urantulkhuur. B,S. W. Suh,J. D. Kang,B. G. Jang,J. H. Kim,S. H. Kim,J. H. Lee,K. D. Choi 한국가금학회 2007 한국가금학회 정기총회 및 학술발표회 Vol.24 No.-

Fowl typhoi d (FT) i s a severe systemic disease of chickens causing heavy economic losses to the poultry industry through mortality, reduced egg production and culling of precious breeding stocks. It is hard to be discovered by clinical sign, pathology and immunology. The immune response to S. gallinarum is poorly characterized because of its virulence. In this study, the total RNA isolated by spleen lymphocytes for Affymetrix GeneChip and RT-PCR analysis. Total RNA was extracted using TRIzol (Invitrogen, Carlsbad, CA). These results indicate that expression of LY86, FN1, K60, CCL19, ah22 increased and during immune response at chickens B-FVI was decreased in fowl typhoid. In conclusion, the differential expressed genes in spleen lymphocytes from S. gallinarum infected and uninfected control groups were related with immunological defense system of chickens. The several identified gene may be used as valuable in formation for immune response mechanims of FT biology.

가금티푸스 감염시 Saccharomyces exiguus를 포함한 사료 첨가제가 산란계의 면역 반응에 미치는 영향

서상원,임희경,김대영,Baatartsogt. O,Urantulkhuur. B,P.K. Mandal,이치호,이준헌,최강덕 한국가금학회 2008 한국가금학회 정기총회 및 학술발표회 Vol.25 No.-

This experiment was conducted to investigate the effects of Saccharomyces exiguss supplementation on mortality and immune response with fowl typhoid infection in layer chicks. A total 162 layer chicks of 10 days age were divided into control and different groups. Mortality was checked daily, Body weight was checked at 0, 3 and 6 day post infection(dpi) and blood was collected at 12 dpi for measuring of IgG and IgA by ELISA. There was no mortality in control groups with or without yeast. The mortality between 6~15 dpi was highest (68~84%) in infected group without yeast. However, the mortality reduced (36~44%) significantly in the groups with yeast orally or by both orally and through feed (20~36%). Weight gain was significantly higher in control groups than the infected groups and there was no difference between the infected groups. IgG level increase in both control and infected chicken when given orally than when given both orally and though feeding. In case of IgM though the level increased in control group but it has gradually decreased in the infected group with the use of yeast. The yeast feeding has reduced mortality of chicks from fowl typhoid, though it is not very effective on weight gain and immune response.