육계 생산성 및 맹장내 미생물에 대한 유산균의 첨가 효과

김상호,박수영,유동조,나재천,최철환,박용윤,이상진,류경선 한국가금학회 2000 韓國家禽學會誌 Vol.27 No.1

This study was carried out to examine the effects of Lactobacillus spp. on performance and cecal microflora in broiler chicks. Eight diets consisting of basal diet(C), C+Lactobacillus crispatus BC7 (T2), C+Lactobacillus reuteri BC9 (T3), C+L.reuteri BC5+L.crispatus BC9 (T4), C+L. reuteri BC5 (T1)+L.reuteri BC9 (T5), C+L.crispatus BC7+L.reuteri BC9 (T6) and C+L.reuteri BC5+L.crispatus BC7+L.reuteri BC9 (T7), were fed to Ross male broiler chicks for 5 weeks. The level of supplemented Lactobacillus spp. was 107 cfu/g diet. Body weight and feed intake were measured every week, and cecal microfla was counted at 1 and 5 weeks for Lactobacillus and yeast. Body weight increased signigicantly in supplemental Lactobacillus treatments from 2 weeks of age (p<0.05). Chicks in T2, T5 and T5 of treatments were heavier than those of other treatments at 5 weeks of age(p<0.05). Viability was not different significantly. Feed intake and feed conversion also were not different, although feed conversion improved slightly in supplemental Lactobacillus treatments. Lactobacillus spp. of cecal content was increased in supplemental Lactobacillus treatments at 1 week, but no significance was found. The number of yeast in cecum was not different from that of supplemental Lactobacillus. In conclusion, Lactobacillus, L.crispatus BC7 and L.reuteri BC9 from broiler cecum could contribute to the increase in body weight with supplemented mono-or mixing of di-lactobacilli. The results indicate that Lactobacillus feeding can benefit to intestinal lactobacillus at early growing broiler.

김치에서 분리한 Lactobacillus sp. JC-7과 Lactobacillus acidophilus 88간의 Electrofusion 최적조건 설정

조영배,최현정,백형석,전홍기 부산대학교 유전공학연구소 1997 분자생물학 연구보 Vol.13 No.-

김치 발효 숙성기간을 연장하고 신선도를 오랫동안 유지 할 수 있는 김치발효 starter의 개발을 목적으로 최적숙성기에 있는 김치로부터 bacteriocin생성능이 없는 유산균을 분리하여 동정하였으며, bacteriocin생성능이 우수한 L.acidophilus 88을 융합시키기 위해 electrofusion에 대한 최적 조건을 검토하였다. 분리균주는 Lactobacillus속으로 동정되어 Lactobacillus sp. JC-7이라 명명하였다. Electrofusion에 의해 생성된 융합주를 식별하기 위해 streptomycin(2.5㎎/ml)에 내성을 나타내는 Lactobacillus sp. JC-7변이주와 kanamycin(600㎍/ml)에 내성을 나타내는 L.acidophilus 88변이주를 분리하였다. Electrofusion을 100V/㎝, 120msec(72ohms,1670 capacitance) 에서 수행했을 때 융합효율이 가장 양호하였으며 전기장의 세기와 시간이 중가할수록 융합 효율이 현저히 감소하였다. 2가 양이온은 농도가 중가할수록 대체적으로 융합효율을 감소시키는 경향을 나타내었으나 1mM MGCl_2에서는 대조군에 비해 융합효율이 약간 증가하였다. PEG매개에 의한 융합법의 융합효율을 비교한 결과, 융합효율은 chemical fusion<electrofusion<electrofusion+20% PEG순을 나타내었다. Evaluation of Optimum Conditions for the Electrofusion between Lactobacillus sp. JC-7 Isolated from Kimchi and Lactobacillus acidophilus 88. Young-Bae Jo, Hyun-Jung Choi, Hyung-Suk Baik and Hong-Ki Jun*. Department of Microbiology, Pusan National University, Pusan 609-735,Korea-A lactic acid bacterium was isolated from kimchi. The isolated strain was identified as the genus Lactobacillus thrugh its morphological characteristics and named as Lactobacillus sp. JC-7. The optimum conditions for the electrofusion between streptomycin(2.5㎎/ml)resistant mutant of Lactobacillus acidophilus 88 and kanamycin(600㎍/ml) resistant mutant Lactobacillus sp. JC-7 were evaluated. The highest number of fusants were obtained at a capacitance value of 120msec(1670㎌), a field strength of 100V/㎝,and a pulse controller setting of 72Ω. The potimum pH of elecroporation buffer was 7.5 and the concentration of divalent cation was 1mM MG^2+. Electrofusants were efficiently obtained by addition 20% polyethylene glycol to electroporation buffer. The yield of fusion was better than that of using polyethylene glycol mediated chemical induction.

유산균의 첨가 급여가 산란 생산성, 소화기관 미생물 변화 및 계란 품질에 미치는 영향

김상호,박수영,유동조,이상진,강보석,최철환,류경선 한국가금학회 2000 韓國家禽學會誌 Vol.27 No.3

A feeding trial was carried out effect of supplemental Lactobacillus on productivity, egg quality and intestinal microflora in 320 21 weeks - old laying hens for 12 week. Supplemented Lactobacillus strains were Lactobacillus amylovorus LLA7(LA), Lactobacillus crispatus LLA9(LC) and Lactobacillus vaginalis LLA11(LV). Three strains mixed to basal diet which containing 2,800㎉/kg ME, 16% CP with none, LA, LC, LV, LA+LC, LA+LV, LC+LV and LC+LC+LV. Supplemental level was 10(sup)7 cfu/g diet. Egg production was tended to increase with adding Lactobacilus, but not difference significantly. Average egg weight was heavier in adding Lactobacillrs compared to the none, and heaviest in LA+LV, LC+LV(P〈0.05). In periodic observation, the gap of egg weight with adding Lactobacillus or not was severe persisting laying periods. The diet containg MC or LV was better than LA, which means the difference by Lactobacillus strains for egg weight. Daily egg mass also increased in adding Lactobacillus about 1.1 to 2.3 g/hen, but not difference significantly. Feed intake and feed conversion were not difference regardless Lactobacillus strains and laying periods. Haugh unit improved with adding Lactobacillus. Cecal Lactobacillus spp. was increased with adding Lactobacillus(P〈0.05), didn't observed E. coli depression. In summary, supplemental Lactobacillus could improve for egg production, egg weight, egg mass and egg white. And those of effect expect much beneficial with mixing Lactobacillus which established well as single strain.

Effect of the Transformed Lactobacillus with Phytase Gene on Pig Production Performance, Nutrient Digestibility, Gut Microbes and Serum Biochemical Indexes

Yin, Q.Q.,Chang, J.,Zuo, R.Y.,Chen, L.Y.,Chen, Q.X.,Wei, X.Y.,Guan, Q.F.,Sun, J.W.,Zheng, Q.H.,Yang, X.,Ren, G.Z. Asian Australasian Association of Animal Productio 2010 Animal Bioscience Vol.23 No.2

In order to improve the availability of phytase and probiotics together, a phytase gene from Aspergillus ficuum has been expressed in Lactobacillus. In this study, the transformed Lactobacillus with phytase gene was fed to pigs to determine its effect on pig production, feed conversion and gut microbes. Forty eight, 60-day-old, castrated pigs (Duroc${\times}$Landrace${\times}$Pietrain) were assigned to 6 groups, 8 pigs for each group. Group 1 was the control, group 2 was added with chlortetracycline (500 mg/kg), group 3 was added with the transformed Lactobacillus (500 mg/kg) with 20% (w/w) of calcium monohydrogen phosphate (CMP, $CaHPO_{4}$) removed, group 4 was added with the natural Lactobacillus (500 mg/kg) with 20% (w/w) of CMP removed, group 5 was added with the transformed Lactobacillus (500 mg/kg) with 40% (w/w) of CMP removed, group 6 was added with phytase (500 mg/kg) with 40% (w/w) of CMP removed. The results showed: i) the average daily gain (ADG) was improved in groups 2, 3 and 4 (p<0.05); ii) the diarrhea rates in the groups added with Lactobacillus were lower than in the other groups (p<0.05), in which the transformed Lactobacillus had more effect on reducing digestive disease; iii) the transformed Lactobacillus was most effective in improving the digestibilities of crude protein (CP), calcium (Ca), phosphorus (P), compared with the other groups (p<0.05); iv) Lactobacillus could increase lactic acid bacterium number and ammonia concentrations, and decrease pH values and E. coli number in pig feces (p<0.05); v) the phytase activity in the feces of pigs fed with the transformed Lactobacillus was 133.32 U/g, which was higher than in group 4 (9.58 U/g, p<0.05), and was almost the same as group 6 (135.94 U/g); vi) the transformed Lactobacillus could increase serum concentrations of IgA, triglyceride, and glutamic oxaloacetic transaminase activity (p<0.05), and had no significant effect on other serum indexes (p>0.05).

김치에서 분리한 Lactobacillus sp. JC-7과 Lactobacillus acidophilus 88간의 Electrofusion 최적조건 설정

조영배,최현정,백형석,전홍기 한국산업미생물학회 1997 한국미생물·생명공학회지 Vol.25 No.2

김치 발효 숙성기간을 연장하고 신선도를 오랫동안 유지할 수 있는 김치 발효 starter의 개발을 목적으로 최적 숙성기에 있는 김치로부터 bacteriocin 생성능이 없는 유산균을 분리하여 동정하였으며, bacteriocin 생성능이 없는 분리균주와 bacteriocin생성능이 우수한 L. acidophilus 88을 융합시키기 위해 electrofusion에 대한 최적 조건을 검토하였다. 분리균주는 Lactobacillus속으로 동정되어 Lactobacillus sp. JC-7이라 명명하였다. Electrofusion에 의해 생성된 융합주를 식별하기 위해 streptomycin (2.5 ㎎/㎖)에 내성을 나타내는 Lactobacillus sp. JC-7 변이주와 kanamycin(600 ㎍/㎖)에 내성을 나타내는 L. acidophilus 88 변이주를 분리하였다. Electrofusion을 100 V/㎝, 120 msec(72 ohms, 1670 capacitance)에서 수행했을 때 융합효율이 가장 양호하였으며 전기장의 세기와 시간이 증가할수록 융합 효율이 현저히 감소하였다. 2가 양이온은 농도가 증가할수록 대체적으로 융합효율을 감소시키는 경향을 나타내었으나 1 mM MgCl_2에서는 대조군에 비해 융합효율이 약간 증가하였다. PEG 4000을 electrofusion buffrer에 20%되도옥 첨가했을 때 가장 양호한 융합효율을 나타내었으며 그 이상에서는 오히려 융합효율이 감소하였다. Electrofusion법과 PEG매개에 의한 융합법의 융합 효율을 비교한 결과, 융합효율은 chemical fusion<electofusion<electrofusion+20% PEG 순을 나타내었다. A lactic acid bacterium was isolated from kimchi. The isolated strain was identified as the genus Lactobacillus through its morphological, cultural, and physiological characteristics and named as Lactobacillus sp. JC-7. The optimum conditions for the electrofusion between streptomycin (2.5 ㎎/㎖) resistant mutant of Lactobacillus acidophilus 88 and kanamycin (600 ㎍/㎖) resistant mutant of Lactobacillus sp. JC-7 were evaluated. The highest number of fusants were obtained at a capacitance value of 120 msec (1670 ㎌), a field strength of 100 V/㎝, and a pulse controller setting of 72Ω. The optimum pH of electroporation buffer was 7.5 and the concentration of divalent cation was 1 mM Mg^2+. Electrofusants were efficiently obtained by addition 20% polyethylene glycol to electroporation buffer. The yield of fusion was better than that of using polyethylene glycol mediated chemical induction.

반응표면분석법을 이용한 Lactobacillus 균주 배양조건의 통계적 최적화

황영민,이희석 한국식품위생안전성학회 2023 한국식품위생안전성학회지 Vol.38 No.5

프로바이오틱스 제품에 대한 수요가 지속적으로 증가하 고 있으며, Lactobacillus 균주가 가장 대중적인 프로바이 오틱스로 널리 사용되고 있다. 프로바이오틱스는 기준에 적합한 균수의 확보가 중요하며 제조원가나 시간 등을 낮 추기 위해 배양법의 개발이 필요하므로 Lactobacillus 생 산을 위한 배양 조건이 최적화되었다. 반응표면방법론에 의한 통계적 최적화에서 반응 변수에 영향을 미치는 독립 변수의 최적 조건은 Lactobacillus acidophilus의 경우 22.55 시간(배양시간), 25o C(배양온도), 3.41%(프리바이오틱스 농 도); Lactiplantibacillus plantarum의 경우 24시간, 30.86o C, 2.00%; Lacticaseibacillus rhamnosus의 경우 66.67시간, 35o C, 3.41%이었다. Lactobacillus의 최적 배양조건은 예측 한 결과와 실제 결과가 밀접하게 일치하는 것을 확인하였 다. 이러한 데이터는 수율 높은 Lactobacillus를 생산하는 데 중요한 포인트를 제공할 것이다. The demand for probiotic products has been steadily increasing, and Lactobacillus strains are widely used and are currently the most popular probiotics. Optimizing culture conditions for Lactobacillus production for use as probiotics will enhance their profitability by reducing production costs and time. Statistical analysis using response surface methodology revealed the following optimal sets of independent variables: 22.55 h (cultivation time), 25°C (cultivation temperature), and 3.41% (w/w, prebiotics concentration) for Lactobacillus acidophilus; 24 h, 30.86°C, and 2% (w/w) for Lactiplantibacillus plantarum; 66.67 h, 35°C, and 3.41% (w/w) for Lacticaseibacillus rhamnosus. Actual outcomes using predicted optimal conditions for Lactobacillus strains have been confirmed to closely match predicted results. This study will provide valuable guidelines for high yield Lactobacillus production.

Development of Species-Specific Primers for PCR Identification of Lactobacillus hilgardii and Lactobacillus farciminis in Kimchi

Myung-Ki Lee,Kyung-Hyung Ku,Young-Jin Kim,Kyung-Hee Kim,Yu-Ri Kim,Hye-Jung Yang 한국식품영양과학회 2010 Preventive Nutrition and Food Science Vol.15 No.2

The aim of this study was to develop species-specific primer sets for kimchi Lactobacillus. Known gene sequences of Lactobacillus 16S rRNA were collected from the NCBI Gene bank, and 69 primer sets were designed using the homologous gene sequence. Six species of kimchi Lactobacilli were used as reference strains: Lactobacillus brevis KCTC3102, Lactobacillus farciminis KCTC3681, Lactobacillus fermentum KCTC3112, Lactobacillus hilgardii KCTC3500, Lactobacillus plantarum KCTC3099, and Lactobacillus sanfranciscensis KCTC3205. PCR amplification and gel electrophoresis were performed to identify the accuracy and specificity of the developed primer set. The results show that the primer set of 5'-aagcctgcgaaggcaag-3' & 5'-aggccaccggctttg-3', 5'-acatactatgcaaatctaagagattagacg-3' & 5'-actgagaatggctttaagagattagcttac-3' resulted in a specific PCR band on L. hilgardii, and primer set of 5'-ctaataccgcataacaactactttcacat-3' & 5'-aacttaataaaccgcctacattctctttac-3' on L. farciminis. The results indicate that the developed primer sets can provide a useful tool for the identification and differentiation of L. hilgardii and L. farciminis from other Lactobacillus species of kimchi.

Development of Species-Specific Primers for PCR Identification of Lactobacillus hilgardii and Lactobacillus farciminis in Kimchi

Lee, Myung-Ki,Ku, Kyung-Hyung,Kim, Young-Jin,Kim, Kyung-Hee,Kim, Yu-Ri,Yang, Hye-Jung The Korean Society of Food Science and Nutrition 2010 Preventive Nutrition and Food Science Vol.15 No.2

The aim of this study was to develop species-specific primer sets for kimchi Lactobacillus. Known gene sequences of Lactobacillus 16S rRNA were collected from the NCBI Gene bank, and 69 primer sets were designed using the homologous gene sequence. Six species of kimchi Lactobacilli were used as reference strains: Lactobacillus brevis KCTC3102, Lactobacillus farciminis KCTC3681, Lactobacillus fermentum KCTC3112, Lactobacillus hilgardii KCTC3500, Lactobacillus plantarum KCTC3099, and Lactobacillus sanfranciscensis KCTC3205. PCR amplification and gel electrophoresis were performed to identify the accuracy and specificity of the developed primer set. The results show that the primer set of 5'-aagcctgcgaaggcaag-3' & 5'-aggccaccggctttg-3', 5'-acatactatgcaaatctaagagattagacg-3' & 5'-actgagaatggctttaagagattagcttac-3' resulted in a specific PCR band on L. hilgardii, and primer set of 5'-ctaataccgcataacaactactttcacat-3' & 5'-aacttaataaaccgcctacattctctttac-3' on L. farciminis. The results indicate that the developed primer sets can provide a useful tool for the identification and differentiation of L. hilgardii and L. farciminis from other Lactobacillus species of kimchi.

Lactobacillus Persisters Formation and Resuscitation

( Hyein Kim ),( Sejong Oh ),( Sooyeon Song ) 한국미생물생명공학회 2024 Journal of microbiology and biotechnology Vol.34 No.4

Lactobacillus is a commonly used probiotic, and many researchers have focused on its stress response to improve its functionality and survival. However, studies on persister cells, dormant cells that aid bacteria in surviving general stress, have focused on pathogenic bacteria that cause infection, not Lactobacillus. Thus, understanding Lactobacillus persister cells will provide essential clues for understanding how Lactobacillus survives and maintains its function under various environmental conditions. We treated Lactobacillus strains with various antibiotics to determine the conditions required for persister formation using kill curves and transmission electron microscopy. In addition, we observed the resuscitation patterns of persister cells using single-cell analysis. Our results show that Lactobacillus creates a small population of persister cells (0.0001-1% of the bacterial population) in response to beta-lactam antibiotics such as ampicillin and amoxicillin. Moreover, only around 0.5- 1% of persister cells are heterogeneously resuscitated by adding fresh media; the characteristics are typical of persister cells. This study provides a method for forming and verifying the persistence of Lactobacillus and demonstrates that antibiotic-induced Lactobacillus persister cells show characteristics of dormancy, sensitivity of antibiotics, same as exponential cells, multi-drug tolerance, and resuscitation, which are characteristics of general persister cells. This study suggests that the mechanisms of formation and resuscitation may vary depending on the characteristics, such as the membrane structure of the bacterial species.

Effect of oral spray with Lactobacillus on growth performance, intestinal development and microflora population of ducklings

Zhang, Qi,Jie, Yuchen,Zhou, Chuli,Wang, Leyun,Huang, Liang,Yang, Lin,Zhu, Yongwen Asian Australasian Association of Animal Productio 2020 Animal Bioscience Vol.33 No.3

Objective: The aim of this study is to investigate the effect of oral spray with probiotics on the intestinal development and microflora colonization of hatched ducklings. Methods: In Exp. 1, an one-way factorial design was used to study the antibacterial activity of the probiotics and metabolites on Escherichia coli (E. coli) without antimicrobial resistance. There were four experimental groups including saline as control and Lactobacillus, Bacillus subtilis, combined Lactobacillus and Bacillus subtilis groups. In Exp. 2, 64-day-old ducklings were allotted to 2 treatments with 4 replicated pens. Birds in the control group were fed a basal diet supplemented with Lactobacillus fermentation in the feed whereas birds in the oral spray group were fed the basal diet and administrated Lactobacillus fermentation by oral spray way during the first week. Results: In Exp. 1, the antibacterial activities of probiotics and metabolites on E. coli were determined by the diameter of inhibition zone in order: Lactobacillus>combined Lactobacillus and Bacillus subtilis>Bacillus subtilis. Additionally, compared to E. coli without resistance, E. coli with resistance showed a smaller diameter of inhibition zones. In Exp. 2, compared to control feeding group, oral spray group increased (p<0.05) the final body weight at d 21 and average daily gain for d 1-21 and the absolute weight of the jejunum, ileum and total intestine tract as well as cecum Lactobacillus amount at d 21. Conclusion: Lactobacillus exhibited a lower antibacterial activity on E. coli with resistance than E. coli without resistance. Oral spray with Lactobacillus fermentation during the first week of could improve the intestinal development, morphological structure, and microbial balance to promote growth performance of ducklings from hatch to 21 d of age.