• Title/Summary/Keyword: Specific pathogen free

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Prophylactic Effects of Bifidobacterium longum HY8001 against Escherichia coli O157:H7 and Salmonella typhimurium DT104 Enteric Infection and Evaluation of Vero Cytotxin Neutralizing Effects (Bifidobacterium longum HY8001 균주의 Escherichia coli O157:H7과 Salmonella typhimurium DT104 장곤 내 감염 예방효과 및 Vero cytotoxin 중화효과)

  • 양수진;윤장원;서근석;구혜정;김소현;배형석;백영진;박용호
    • Microbiology and Biotechnology Letters
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    • v.27 no.5
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    • pp.419-425
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    • 1999
  • Prophylactic effects of Bifidobacterium longum HY8001, Korean isolate, against Escherichia coli O157:H7 and Salmonella typhimurium DT104 enteric infection were examined at four groups of specific pathogen free(SPF)-ICR mouse for each pathogen. B. longum HY8001+B. typhimurium DT104+B. longum HY8001(BL+ST+BL) group and B. longum HY8001+E. coli O157:H7+B. longum HY8001(BL+E+BL) group were fed with B. longum HY8001 before and after E. coli O157:H7 or s. typhimurium DT104 challenge, while B. longum HY8001+S. typhimurium DT104(BL+ST) and B. longum HY8001+e. coli O157:H7(BL+E) groups were fed with B. longum HY8001 only before E. coli O157:H7 or S. typhimurium DT104 challenge. E. coli O157:H7(E) and S. typhimurium DT104(ST) groups were challenged with each pathogen without B. longum HY8001 administration and control groups were administered with phosphate buffered solution(PBS). After the oral administration with B. longum HY8001(109cfu), th emice were challenged with E. coli O157:H7(2$\times$1010cfu) or S. typhimurium DT104(108cfu) and the mortality rate and the fecal shedding of challenged pathogen were also examined define the reactivity of the B. longum HY8001. Production of toxin neutralizing substance(s) of B. longum HY8001 was determined by cell cytotoxicity assay using Vero cells. Fecal shedding of th eS. typhimurium DT104 was significantly decreased in BL+ST+BL group fed with B. longum HY8--1 before and after challenge(p<0.05), while the fecal shedding s of S. typhimurium DT104 in BL+ST and St groups remained more than 106cfu. the protective effect of the B. longum HY8001 against E. coli O157:H7 was significantly high only in BL+E+BL group fed with b. longum Hy8001 before and after E. coli O157:H7 challenge from the result of fecal E. coli O157:H7 isolation rate, mortality rate, and intestinal contents culture to detect E. coli O157:H7. the mortality rate of the BL+e and E groups. The cytopathic effect (CPE) of the Vero cytotoxin (Shiga like toxin I & II) in Vero cell was neutralized in B. longum HY8001 culture supernatant added wells which indicate the presence of soluble Vero cytotxin neutralizing substance(s) in B. longum HY8001 culture suprnatant.

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Characterization analysis of Rongchang pig population based on the Zhongxin-1 Porcine Breeding Array PLUS

  • Dong Leng;Liangpeng Ge;Jing Sun
    • Animal Bioscience
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    • v.36 no.10
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    • pp.1508-1516
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    • 2023
  • Objective: To carry out a comprehensive production planning of the existing Rongchang pig population from both environmental and genetic aspects, and to establish a closed population with stable genetic diversity and strict pathogen control, it is necessary to fully understand the genetic background of the population. Methods: We genotyped 54 specific pathogen free (SPF) Rongchang pigs using the Zhongxin-1 Porcine Breeding Array PLUS, calculated their genetic diversity parameters and constructed their families. In addition, we also counted the runs of homozygosity (ROH) of each individual and calculated the value of inbreeding coefficient based on ROH for each individual. Results: Firstly, the results of genetic diversity analysis showed that the effective population size (Ne) of this population was 3.2, proportion of polymorphic markers (PN) was 0.515, desired heterozygosity (He) and observed heterozygosity (Ho) were 0.315 and 0.335. Ho was higher than He, indicating that the heterozygosity of all the selected loci was high. Secondly, combining the results of genomic relatedness analysis and cluster analysis, it was found that the existing Rongchang pig population could be divided into four families. Finally, we also counted the ROH of each individual and calculated the inbreeding coefficient value accordingly, whose mean value was 0.09. Conclusion: Due to the limitation of population size and other factors, the genetic diversity of this Rongchang pig population is low. The results of this study can provide basic data to support the development of Rongchang pig breeding program, the establishment of SPF Rongchang pig closed herd and its experimental utilization.

SPF 닭에서 재조합 H9N3 조류 인플루엔자 백신의 효능과 안전성 평가

  • Sin, Jeong-Hwa;Mo, In-Pil
    • Proceedings of the Korea Society of Poultry Science Conference
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    • 2006.11a
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    • pp.90-91
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    • 2006
  • To reduce the economic impact and control Low pathogenic avian influenza (LPAI), vaccination with inactivated vaccine has been considered in this country. We tried to develop inactivated vaccine with reassorted H9N3 AI virus which has different type of neuraminidase compare to those of field AI virus. Before reassorted vaccine was produced, we confirm the virus as master seed by limiting dilution, RT-PCR and sequencing method. Also, we evaluate the biological characteristics of the virus to find out the possibility of prevention against field infection of AI virus. Finally, we evaluate the safety and efficacy of the vaccine made of reassorted AI virus in the specific pathogen free (SPF) chickens. After limiting dilution, we choose RV7CE4 as a vaccine candidate and compare the gene sequence of this vaccine strain to those of AI05GA which is parents strain. Compared to amino acid sequences of specific gene of AI05GA and RV7CE4, exhibited a high degree of amino acid sequence homology. In the safety and efficacy test, there were no specific clinical signs or mortality. Reassorted H9N3 viruses were reisolated in cloaca swab on 5 days post inoculation. In the vaccine study, once or twice vaccination was performed and challenged with H9N2 field virus (01310). Vaccine has no adverse effect on birds and formed good immune capability which reduce viral shedding in the birds infected with 01310. Based on the above result, we developed reassorted H9N3 vaccine which will efficiently prevent the low pathogenic AIV (H9N2) infection in the poultry farms.

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The Use of the Pathogen-specific Bacteriophage BCP8-2 to Develop a Rice Straw-derived Bacillus cereus-free Starter Culture (단일 박테리오파지를 이용한 볏짚 유래 Bacillus cereus free 스타터 컬쳐의 개발)

  • Bandara, Nadeeka;Chung, Seo-Jin;Jeong, Do-Youn;Kim, Kwang-Pyo
    • Korean Journal of Food Science and Technology
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    • v.46 no.1
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    • pp.115-120
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    • 2014
  • The purpose of this study was to develop a rice straw-derived Bacillus cereus (B. cereus)-free starter culture for traditional soybean fermented products using a B. cereus-specific bacteriophage, BCP8-2. To determine the optimal medium that supports the growth of rice straw-derived microorganisms and BCP8-2 activity, 5 different culture media were tested. The 5% ground bean (GB) medium was selected for further study. No B. cereus was detected in the BCP8-2-treated rice straw in GB medium, whereas B. cereus at a level of $10^7$ CFU/mL was recovered in the no-phage control. The total bacterial count reached approximately $10^9$ CFU/mL regardless of phage addition. When the 16S rRNA sequence-based microbial community was monitored using denaturing gradient gel electrophoresis (DGGE) and pyrosequencing, a similar microbial community was observed in the phage-treated and control samples. In conclusion, we demonstrate that phage can be used to prepare a rice straw-derived B. cereus-free starter culture with minimal effect on natural microflora.

Phenotypical changes of lymphocyte subsets infiltrated in the skin lesions induced experimentally by very virulent strain of Marek's disease virus in chickens (마렉병 바이러스 강독주의 실험 접종에 의해 유발된 닭 피부병변에 침윤한 림프구 표현형의 변화)

  • Cho, Kyoung-Oh
    • Korean Journal of Veterinary Research
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    • v.41 no.3
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    • pp.373-380
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    • 2001
  • Marek's disease virus (MDV) can cause skin lesions including inflammatory to tumorous. The phenotypical changes of lymphocytes infiltrating in the skin lesions induced by MDV were not clear. Therefore, the skin biopsies taken at weekly intervals for 8 weeks from the same specific-pathogen free chickens inoculated with Md/5 MDV were examined to analysis the phenotypical changes of lymphocytes. Histologically skin lesions progressed from initial inflammatory to late tumorous. Sequentially CD4+ T lymphocytes increased gradually in number from initial skin lesions and were major composition cells in the tumor lesions. Regardless of inflammatory or tumor lesions, CD8+ T cells and ${\gamma}{\delta}$ T cells infiltrated particularly in the dermis and subcutaneous on which MDV was actively replicated in the feather follicle epithelium(FFE). In addition, IgG bearing B lymphocytes in considerable number infiltrated in the dermis and subcutaneous tissues. From these results, the development of MDV-induced skin lesions was inflammatory following tumorous. In addition, each CD8+, ${\gamma}{\delta}$ and CD4+ T cells and B cell might act to protect MDV replication in the FFE or tumor cells which turned on lytic cycle.

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The Probiotic and Adherence Properties of Lactobacillus reuteri Pg4 Expressing the Rumen Microbial β-Glucanase

  • Yu, B.;Liu, J.R.;Hsiao, F.S.;Lee, T.T.;Chiou, P.W.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.9
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    • pp.1324-1329
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    • 2008
  • This study was conducted to evaluate the potential of the transformed Lactobacillus reuteri Pg4 (T-Pg4) harboring the ${\beta}$-glucanase gene as a poultry probiotic. The probiotic properties of the T-Pg4 strain were evaluated in vitro by their adherence capability and acid and bile salt tolerance, and were evaluated in vivo by their survival and adhesion in the gastrointestinal tract (GIT) of specific-pathogen-free (SPF) chickens. The results showed that the T-Pg4 strain exhibited resistance to acidic conditions and contact with bile salt, and adhered efficiently to the crop and intestinal epithelial cells of chickens in vitro. The T-Pg4 strain also could survive and colonize the gastrointestinal epithelium of the experimental SPF chickens in vivo. In addition, radial enzyme diffusion was used to demonstrate that the Lactobacillus spp. randomly isolated from the GIT of the SPF chickens fed T-Pg4 possessed ${\beta}$-glucanase secretion capability. These findings have demonstrated that the transformed L. reuteri Pg4 survives transit through the stomach and intestine, and may secrete ${\beta}$-glucanase in the chicken GIT. Therefore, it is suggested that this organism could be used as a multifunctional poultry probiotic.

Genotoxicity Test of Bojungikkeehapdaechilki-tang water extract (보중익기합대칠기탕(補中益氣合大七氣湯) 추출물의 유전독성 평가)

  • Hwang, Hui-Jeung;Byun, Joon-Seok;Heo, Jin-Il
    • Herbal Formula Science
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    • v.14 no.1
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    • pp.141-167
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    • 2006
  • The genotoxicity of water extract of Bojungikkeehapdaechilki-tang was tested by In Vitro Chromosome Aberration Test. Bacterial Reverse Mutation Assay and Micronucleus test according to OECD Guidelines and KFDA Guidelines. The obtained results were as follows : 1. Chromosome Aberration Test: In Vitro Chromosome Aberration Test of Bojungikkeehapdaechilki-tang extracts was carried out using cultured Chinese hamster lung cells in the presence and absence of metabolic activation system(S-9 mix). No significant changes in the number of aberrant metaphases having structural and number of aberrations were detected in Bojungikkeehapdaechilki-tang extracts treated groups. 2. Bacterial Reveres Mutation Assay: Bojungikkeehapdaechilki-tang extracts was evaluated for its potential to induce reverse mutation in the histidine auxotroph strains of Salmonella typhimurium such as TA100, TA1535, TA98 and TAl537 and the tryptophan auxotroph strain of Escherichia coli WP2 uvrA. No significant changes in the number of revertant colonies compared to its negative control were detected in Bojungikkeehapdaechilki-tang extracts treated groups against all 5 strains. 3. Micronucleus test: Micronucleus test of Bojungikkeehapdaechilki-tang extracts were performed using specific pathogen free 7-week old male ICR mouse. No significant changes in the number of micronucleated polychromatic erythrocytes among 2000 polychromatic erythrocytes compared to negative control were detected in all Bojungikkeehapdaechilki-tang extracts treated groups. In summarized above-mentioned results, it is concluded that Bojungikkeehapdaechilki-tang extracts have not genotoxicity against In Vitro Chromosome Aberration Test, Bacterial Reverse Mutation Assay and Micronucleus test.

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Toxicity Study of Streptococcus pneumoniae Vaccine Administrated Subcutaneously in Rats

  • Park, Sin-Jeong;Seo, Kook-Heon;Han, Sang-In
    • Toxicological Research
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    • v.27 no.2
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    • pp.111-118
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    • 2011
  • This study was performed to evaluate the toxicity of polysaccharide-based Streptococcus pneumoniae vaccine in Specific Pathogen Free (SPF), Sprague-Dawley (SD) rats. S. pneumoniae vaccine was administrated subcutaneously each dose level of high (560 ${\mu}g$/rat), medium (280 ${\mu}g$/rat) and low (140 ${\mu}g$/rat) on days 0, 14, 28. The rats were observed for 2 weeks or 4 weeks after the final injection. During this test, there were no significant dose-dependent changes in body weight, water and food consumption. In urinalysis and serum chemistry, dose-related changes were not detected. In hematology, the percent of neutrophils and lymphocytes in white blood cells were changed significantly. According to the measurement of organ weight, only spleen weight was significantly increased in all groups of administration compared to the control group. In the histopathological examination, an antigen-deposit, vacuolated macrophages, infiltrated inflammatory cells and a formation of granulation tissue were observed at the site of an administration. These results are considered as an outcome by immune responses through a vaccination. Consequently, the results of this study demonstrated that S. pneumoniae vaccine has no toxicity when it was administrated subcutaneously three times in 2-week interval at a high dose of 560 ${\mu}g$/rat.

Metabolism of Ginseng Saponins by Human Intestinal Bacteria (Park II) (사람의 장내세균에 의한 인삼 사포닌의 대사(제2보))

  • Hasegawa, Hideo;Ha, Joo-Young;Park, Se-Ho;Matumiya, Satoshi;Uchiyama, Masamori;Huh, Jae-Doo;Sung, Jong-Hwan
    • Korean Journal of Pharmacognosy
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    • v.28 no.1
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    • pp.35-41
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    • 1997
  • Following ginsenoside-Rb1-hydrolyzing assay, strictly anaerobic bacteria were isolated from human feces and identified as Prevotella oris. The bacteria hydrolyzed ginsenoside Rb1 and Rd to $20-O-{\beta}-D-glucopyranosyl-20(S)-protopanaxadiol$ (I), ginsenoside Rb2 to $20-O-[{\alpha}-L-arabinofuranosyl (1{\rightarrow}6)-{\beta}-D-glucopyranosyl] - 20(S)-protopanaxadiol$ (ll) and ginsenoside Rc to $20-O-[{\alpha}-L-arabinofuranosyl (1{\rightarrow} 6){\beta}-D-g1ucopyranosyl]-20(S)-protopanaxadiol$ (III) like fecal microflora, but did not attack ginsenoside Re nor Rgl (Protopanaxatriol-type). Pharmacokinetic studies of ginseng saponins was also performed using specific pathogen free rats and demonstrated that the intestinal bacterial metabolites I-111, 20(S)- protopanaxatriol(IV) and 20(S)-protopanaxadiol(V) were absorbed from the intestines to $blood(0.4-5.1\;{\mu}g/ml)$ after oral administration with total saponin(1 g/kg/day).

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Ocular Lesions Induced Experimentally by Very Virulent Strain of Marek′s Disease Virus in Chickens (닭의 마렉병 바이러스 강독주 실험접종에 의한 안구병변)

  • Cho, Kyoung-Oh
    • Korean Journal of Veterinary Pathology
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    • v.5 no.2
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    • pp.49-56
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    • 2001
  • Ocular lesions induced in 40 specific-pathogen-free Marek's disease (MD) resistant chicks by inoculation at 1 day of age with very virulent strain of Marek's disease virus (WV) were pathologically examined. Grossly,24/40 (60%) chicks had white gel-like materials in the vitreous body, whereas thickening and discoloration of iris (gray eye) were not observed. Microscopically, characteristic ocular MD lesions were observed in choroid (27/40), ciliary (30/40) and iris (23/40) in which small focal inflammatory to diffuse neoplastic Iymphoid cells were infiltrated. Five out of 40 MDV-inoculated birds revealed necrotizing Iymphomas in choroid. These lesions consisted of necrotic and degenerating Iymphoblasts accompanied by intranuclear inclusion body. There was retinal atrophy and necrosis with inclusion body detected in necrotic ganglion, inner or outer nuclear and infiltrated Iymphoblast cells. Conjunctiva showed lymphoid cell infiltration in 29/40 chicks inoculated with MDV, Vitreous body exhibited mild to severe exudation of eosinophilic proteinaceous material in 24/40 chicks. These lesions were associated with Iymphoid cell infutration, edema and fibrosis of choroid. Pecten (7/40) and optic nerve (13/40) were infiltrated usually mildly with Iymphoid cells. From these results, very virulent strain, Md/5 of MDV caused high incidence of ocular lesions in MD resistant chicks. In addition, Md/5 induced exudation of proteinaceous material into the vitreous body and fibrosis of choroid. Necrotizing ocular Iymphoma lesions in choroid is the first report in the MD literature.

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