프로바이오틱스는 다양한 환경에서 분리되어 왔으며, Bacillus 균주는 유산균과 같은 일반적인 프로바이오틱스의 한계를 보완할 수 있는 내생포자 형성 능력으로 인해 프로바이오틱스 균주 중에서 유리하다. 본 연구의 목적은 한국에서 오염되지 않은 환경으로 알려진 울릉도 토양에서 분리된 Bacillus 균주의 프로바이오틱스 잠재성을 조사하는 것이다. 토양 시료는 울릉도 각지에 서 채취하였으며, 항생제 내성과 효소 활성을 평가하였다. 항생제에 내성을 나타내지 않는 6개 의 Bacillus 균주를 확보하였고 이후에 효소 활성을 검사하였다. 인체에 악영향을 미칠 수 있 는 β-glucuronidase 효소는 활성이 나타나지 않았으며, 시중에서 판매되고 있는 Bacillus 프로바 이오틱스와 유사한 효소의 활성과, 추가적으로 펩타이드 가수분해효소인 Leucine arylamidase 의 활성이 나타난 NB-1 균주가 확인되었다. 이에, 추가적인 효능 연구 및 안정성 실험을 통해 프로바이오틱스 균주로서의 가능성을 모색해야 할 필요가 있다고 판단된다.

A novel L. harbinensis VF was isolated from fermented vinegar and identified through biochemical characteristics and 16S rRNA sequencing analysis. Characteristics of probiotics were studied for acid and bile salt tolerances, hemolytic activity, antibiotic resistance, antimicrobial activity, cell surface hydrophobicity, and aggregation. The survival rates of the isolate were maintained at 68.9% and 95.6% after 3 h incubation at pH 2.0 and 2.5 and were over 74% at bile salt concentrations of 0.3% and 0.5%. The hemolytic activity was confirmed to be -hemolysis. The isolate showed broad antibiotics-resistance in over 12 antibiotics except for trimethoprim/sulfamethoxazole, compared to L. plantarum (KCTC 3108), used as control, and previously reported L. harbinensis. Antimicrobial activity was confirmed against pathogens, B. cereus, E. coli O157:H7, L. monocytogenes, and S. typhimurium, except S. aureus, and it was attributed to produced organic acids (low pH). The hydrophobicity for xylene and chloroform were 16.7% and 20.4%, respectively, lower than those of L. plantarum (41.2% & 49.9%). The auto-aggregation and coaggregation with pathogens were 83.8% and 49.2-87.6% at 24 h incubation, respectively, higher than those of L. plantarum. From this study, L. harbinensis VF is likely to be used as a probiotic strain and is a strong candidate for commercial application.

This in vitro study investigated the enhancement of rumen bacterial adhesion on a substrate to increase the digestibility of rice straw in Hanwoo cattle. The rice straw was pretreated with enzymes, probiotics, or ammonia, and the effects on the enhancement of bacterial adhesion and fiber degradation were analyzed using in vitro rumen fermentation. Enzyme treatment included spraying of cellulase and xylanase at 40 and 120 U per g of rice straw, respectively; Saccharomyces cerevisiae culture of 1.0×107 CFU was sprayed as a probiotic treatment per gram of rice straw; ammonia was sprayed at 0.3% per gram of rice straw. Following enzyme treatment, Fibrobacter succinogenes formed a higher adhesion colony than the control group (7.26±0.03 and 8.43±0.20) after 6h and 12h of in vitro culture (p<0.05), respectively. Attachment of Ruminococcus flavefaciens also increased following enzyme treatment (p<0.05) after 6 and 12 h compared to that of the control (5.18±0.06 and 6.60±0.15); and R. albus attachment showed a significant increase compared to that of the control (5.94±0.15) after 6 h of incubation (p<0.05). Probiotic treatment increased attachment of F. succinogenes in comparison with untreated rice straw after 6 h and 12 h of fermentation (p<0.05); R. flavefaciens attachment showed an increase only after 6 h of culture (p<0.05); R. albus was not affected. Attachment of F. succinogenes, R. flavefaciens, and R. albus increased with ammonia treatment after 6 h and 12 h (p<0.05). Dry matter digestibility was higher after the enzyme treatment (3.45±0.21 and 7.04±0.09) than in the control group(1.85±0.08 and 3.94±0.04) after 6 and 12 h of in vitro culture (p<0.05), respectively. It was also higher than that of untreated rice straw after probiotic and ammonia treatments (p<0.05). There was an increase in the enhancement levels of bacterial adhesion depending on the type of fibrolytic bacteria following enzyme, probiotic, and ammonia treatment. These treatments improved digestibility. This enhancement is considered to be greater following enzyme and ammonia treatments than with probiotics.

The efficacy of standard therapeutic strategies for Helicobacter pylori infection is decreasing over time due to the emergence of drug-resistant strains. At the our previous study, we reported that the lactic acid bacterium Lactobacillus paracasei HP7 isolated from kimchi - a fermented vegetable dish widely consumed in Korea - had inhibitory effects against H. pylori in vitro and in vivo. In this study, we are aimed to investigate the synergistic effect of probiotic HP7 and the extracts of Perilla frutescens (PFE) and Glycyrrhiza glabra (GGE) to alleviate Helicobacter pylori-induced damages. H. pylori infection in KATO III cells decreased mucin gene expression over time. The reduction of mucin gene expression was significantly recovered at 6 hours when GGE, PFE, and Lactobacillus HP7 were co-treated. H. pylori infection of C57BL/6 mice was demonstrated by H. pylori-IgG concentration, and results showed significant increase in mice post-infected with H. pylori. Blood Tumor Necrosis Factor (TNF)-α levels were elevated 4 weeks after H. pylori inoculation. However, the treatment of a combined mixture of HP7 including GGE and PFE induced significant decrease of TNF-α level (p<0.01) and showed significant decrease of TNF-α level as compared as the level by the positive control Glycyrrhiza glabra acetone extract (Deglycyrrhizinated Licorice, DGL) treatment (p<0.05). Additionally, the treatment of a combined mixture of HP7 including GGE and PFE decreased gastric inflammation and mucosal lesions in the stomach of H. pylori-infected mice. These results demonstrate that he treatment of a combined mixture of HP7 including GGE and PFE treatment can inhibit H. pylori growth and is thus a promising treatment for patients with gastric symptoms such as gastritis that are caused by H. pylori infection.

The purpose of this study was to analyze the effects of the pre-treatment method on the measurement of probiotic cell counts. The probiotic cell count was not significantly different in the pre-treatment method such as experimenters, diluted solution, medium, and homogenization duration. The mean value of probiotic cell count with capsule was 2.2×1010±9.5×109 CFU/g. This probiotic cell count was converted into 2.8×1010±1.2×1010 CFU/g based on the net weight. The mean value of probiotic cell count without capsules was 4.3×1010±1.8×1010 CFU/g. As a result of this comparison, probiotic cell count showed significant difference with and without capsules. Thus, it is suggested that the probiotic cell count is measured by removing the capsule in capsule probiotics.

브로일러의 생산성과 닭고기의 품질에 미치는 Bacillus. subtillus, Streptomyces galilaeus 및 Sphingobacteriaceae로 구성된 사료용 복합생균제(PFM, probiotic feed mixture)의 효과를 조사하였다. 병아리 (Ross 308) 240마리를 4개의 처리구로 나누어 35일간 사육하였다. 처리구는 사료용 복합 생균제를 함유하지 않은 대조구(CON, control), 일반생균제(CP3, commercial probiotics 0.3%), 사료용 복합 생균제 3(PFM3, 사료용 복합 생균제 0.3%), 사료용 복합 생균제 5(PFM5, 사료용 복합 생균제 0.5%)로 구분하였다. 체중은 CON과 비교할 때 CP3, PFM3, PFM5가 높았고 PFM3, PFM5는 CP3에 비해 우수하였다(p<0.05). F낭, 흉선, 비장의 무게 및 IgG 수준은 CON과 비교할 때 PFM3, PFM5 및 CP3에서 증가하였으며 PFM3, PFM5는 CP3에 비해 높았다(p<0.05). 맹장의 Lactobacillus 균수는 PFM3, PFM5 및 CP3를 섭취한 브로일러가 CON에 비해서 높았으나 E.coli, Salmonella, coliforms, 총 호기성 균수는 감소하였다(p<0.05). PFM3 및 PFM5를 섭취한 닭가슴살의 보수력은 CON 및 CP3와 비교할 때 증가하였다(p<0.05). 결론적으로 Bacillus subtillus, Streptomyces galilaeus 및 Sphingobacteriaceae를 포함하는 사료용 복합생균제 0.3%가 브로일러의 생산성과 닭고기 품질향상에 기여할 수 있음을 나타낸다.

본 연구는 계란 생산성, 계란품질 및 악취저감에 관한 Bacillus subtillus, Streptomyces galilaeus 및 Sphingobacteriaceae로 구성된 복합생균제(PM, probiotic mixture)의 효과를 조사하였다. 산란계 Hy-Line Brown 240수를 4처리구로 구분하여 6주 동안 사육하였다. 처리구는 복합생균제를 함유하지 않은 대조구(CON, control), 일반생균제(CP3, commercial probiotics 0.3%), 복합생균제 3(PM3, 복합생균제 0.3%), 복합생균제 5(PM5, 복합생균제 0.5%)로 구분하였다. 산란율, 계란 호우유니트, 난각두께, 난각강도, 난황색 및 농후난백 높이는 PM3, PM5가 CON 및 CP3에 비해서 높았다. 계분으로부터 암모니아, E. coli, 총호기성균, Coliform, Salmonella 균수는 CON>CP3>PM3>PM5 순서로 증가하였다. 본 결과는 B. subtillus, S. galilaeus 및 Sphingobacteriaceae로 구성된 복합생균제 0.3%가 산란율, 계란품질 개선 및 분에서 암모니아 발생을 낮추어 양계장 악취저감 효과를 갖는다는 점을 보여 준다.

The purpose of this study was to investigate probiotic characteristics and immune enhancing effects of Bifidobacterium (B.) longum KBB1-26 and BIF-4, B. breve KBB5-22 isolated from human intestine for probiotic use in humans and animals. We measured acid, bile and heat tolerance, antimicrobial activity against pathogenic bacteria, Escherichia (E.) coli, Salmonella (S.) Enteritidis, Staphylococcus (S.) aureus, and Listeria (L.) monocytogenes. Immune enhancing effects of B. longum and B. breve were investigated by measuring nitric oxide (NO), nuclear factor (NF–κb), interleukin-1ß (IL-1ß), interleukin-6 (IL-6), interleukin-12 (IL-12) and tumor necrosis factor- α (TNF- α) in RAW 264.7 cells or RAW BLUE cells. B. longum KBB1-26 was survived at pH 2.0. B. longum KBB1-26 and BIF-4, B. breve KBB5-22 also showed tolerance to 0.3% of oxgall bile salt. B. longum KBB1-26 was able to survive at 70 ℃ and 80 ℃ for 20 min. KBB1-26 showed the antimicrobial inhibition zone to pathogenic bacteria such as E. coli (12 mm), S. Enteritidis (14 mm), S. aureus (14 mm) and L. monocytogenes (41 mm). The production of NO (4.5 ± 0.00μM/mL) and IL-1β (39.7 ± 0.55pg/mL) of KBB1-26 significantly higher than BIF-4 and KBB5-22, respectively. In addition, KBB1-26 and KBB5-22 induce the production of high level of TNF-α and IL-6 in macrophages. Collectively, B. longum KBB1-26 have acid, bile, heat tolerance, antimicrobial activity and immune enhancing effects. These results suggest that KBB1-26 can be used as probiotics for humans and animals.