Biofilms are complexly structured communities of microorganisms composed of surface-attached microorganisms, where their effects on the host have been controversial. In this study, we investigated the potential biofilm-forming capacity of Lacticaseibacillus rhamnosus LRH020 (DSM25568) by detecting genes known to promote biofilm formation. It was shown that the aggregation substance gene (asa 1) was presented in the LRH020 strain. Therefore, we investigated the phenotypic activities of the gene asa1 via two methods: biofilm formation and autoaggregation activity. It was shown that the strain LRH020 had significantly less ability to form biofilm compared to the positive control strain Enterococcus faecalis ATCC 19433. Furthermore, LRH020 exhibited biofilm-forming activity comparable to Lacticaseibacillus rhamnosus GG (LGG), widely used probiotics. The auto-aggregation activity of LRH020 was also within the safe range similar to that of LGG. In conclusion, this study shows that both biofilmforming and auto-aggregation activities of the LRH020 are comparable to one of the most studied probiotics strains, LGG.

Probiotics are live microorganisms that confer health benefits onto the host when administered at adequate doses. Most widely used probiotics, such as lactobacilli and bifidobacteria, are known to be elements of healthy gut microflora and hence are not considered a threat to the host. However, probiotics may pose a risk in certain populations with compromised immune systems or defects in gut barrier functions. Herein, we evaluated the safety of Bifidobacterium breve BB077, according to the safety evaluation guidelines for probiotics produced by the National Institute of Food and Drug Safety Evaluation (NIFDS). The results show that B. breve BB077 is both non-hemolytic and non-cytolytic. In contrast, B. breve BB077 exhibited higher streptomycin and tetracycline resistance than the suggested NIFDS standard cut-off values. Hence, a genetic analysis of the streptomycin and tetracycline resistance genes was performed to determine the origin of antimicrobial resistance. Streptomycin and tetracycline resistance was shown have arisen from chromosomal mutations and considered intrinsic to the taxonomic group. In conclusion, the B. breve BB077 strain might be safe for human consumption.

The detection of the genome-based antibiotic resistance gene is an essential analysis process for the purpose of verifying the safety of probiotic strains, including lactic acid bacteria. In this study, 4 analysis platforms (AMRFinderPlus, staramr, rgi, ABRicate) were used for cross-comparison of 782 genomes corresponding to 19 kinds of probiotic species notified as functional foods. As a result of the analysis, the relatively fewest number of antibiotic resistance genes were detected in strains belonging to the order Lactobacillales, and antibiotic resistance genes were detected in 322 genomes used in the case of 2 types of Enterococcus genus. In addition, the presence and type of antibiotic resistance gene detection showed a lot of difference even for the same genome sequence depending on the database and analysis algorithm used by the analysis platform. These results can be confused in evaluating the potential for transmission of antibiotic resistance genes inherent in specific lactic acid bacteria and predicting potential risks that may occur in the future. Accordingly, it is judged that the antibiotic resistance gene-related analysis criteria need to be established more clearly and specifically in the safety evaluation of probiotic bacteria.

This mini-review focused on the current advances in the development and the application of the antibacterial agents produced from probiotics strains against foodborne pathogens. Inhibitory activity of the pathogen growth could be achieved by co-culture with probiotics and/or the treatment of metabolites extracted from probiotics culture, whereas strain-dependent efficacy was mainly reported according to the bacterial species of the target pathogens. To overcome the limited antibacterial spectrum and the efficacy of the metabolites from probiotics, the recent research highlight the discovery of novel bioactive substances with broad range of the inhibitory activity of foodborne pathogens and mode-of-action which has not been reported as the major research goal. Moreover, understanding the distribution of functional and regulation genes coding the production of the antibacterial metabolites based on the bacterial genome analysis can provide the clues for the mechanisms of the pathogen control by using probiotics. Major strategies on the application of the genomics in this research area can be represented as follows: 1) functional annotation specialized for antimicrobial proteins, 2) assessment of the antibacterial effects followed by the general/functional annotation, 3) genome and metabolites analysis for the purification of antimicrobial proteins, 4) comparative genomics and the characterization of antimicrobial potential. Especially the relationship between phenotype-genotype based on genomic bioinformation with the results of the practical efficacy tests of bacterial metabolites as an active substance of disinfectants and/or preservatives can be expected to act as supportive research for broadening our knowledge regarding the key metabolic pathways associated with the production of antibacterials from probiotics.

이상의 결과를 종합하면, 혼합유산균 2종(Bifidobacterium animalis ssp. Lactis Bf141와 Lactobacillus rhamnosus Lb102)의 섭취는 고지방식이 유도 비만 마우스에서 체중, 체지방, 제지방, 골밀도 등 주요 체성분에 영향을 주지 않았다. 혼합유산균은 식욕 조절 효과를 위해 측정한 사료 섭취량에도 영향을 주지 않았으며, 간 조직 무게에도 영향을 미치지 않았다. 인슐린저항성과 포도당신생합성의 주요 지표인 공복혈당량 또한 혼합유산균 급여에 의해 변화 하지 않았다. 또한 혼합유산균은 심혈관질환의 지표로 사용되는 혈중 중성지방 및 총 콜레스테롤에 영향을 주지 않았으며, 체내 지방의 소화 및 흡수율에서도 영향을 미치지 않았다. 결론적으로, 고지방식이유도 비만 마우스를 이용하여 혼합유산균(Bf141 + Lb102)의 항비만 생리활성 을 검증한 결과, 유의미한 물리적, 대사적 표현형 개선은 검증되지 않았다. 따라서, 추후 개별 보다 다양한 혼합 조건 및 농도로 연구를 설계하여 혼합유산균의 항비만 효과를 검증할 필요성이 있다.

This study investigated various levels of sodium nitrite and probiotics (SNPro) combination as an alternative to zinc oxide on the growth performance, immune response, intestinal microflora, and morphology of weaned pigs. One hundred and ninety-two weaned pigs (Landrace×Yorkshire×Duroc) with an average body weight of 6.51±0.15 kg were randomly assigned to four treatments(n=6) on the basis of their initial body weight. Experimental period was divided into phase 1 and 2 (each 14 days). The dietary treatments were: 1) Basal diet (control), 2) SNPro1 (control+0.01% SNPro), 3) SNPro2 (control+0.02% SNPro), 4) SNPro3 (control+0.03% SNPro). The average daily gain when SNPro was added to the diet was 288, 309, 319, 324 g in phase 1, 355, 387, 410, 407 g in phase 2 and 321, 348, 364, 366 g in the overall. The concentration of interleukin-8 and interleukin-10 in serum when SNPro was added to the diet were 15, 13.5, 13, 12.8 ng/ml and 165, 162, 155, 145 ng/ml (p<0.05) but toll-like receptor 4 and immunoglobulin G levels in serum were no significantly different. The colonization of Escherichia coli in the ileum and Salmonella spp. in the caecum were significantly decreased as SNPro level increased (p<0.05). However, the population of Lactobacillus spp. did not differ among the groups. Although villus height and villus height to crypt depth ratio were not significantly affected by the treatments, crypt depth in the jejunum was 599, 586, 615, 599 ㎛ as SNPro level increased (p<0.05). In conclusion, SNPro had beneficial effects on growth performance, immune response, intestinal microflora and morphology weaned pigs. Therefore, SNPro not only can be considered as an alternative for the pharmacological level of zinc oxide in weaning pigs but also ideal dietary SNPro level was 0.02%.

This research aimed to investigate the effects of the inoculation method and diverse ingredients on the quality properties and probiotics viability of pudding with milk and/or soymilk. The probiotic strain (Lactobacillus acidophilus, LA-5) was inoculated into the yogurt base and fermented until the pH is 5.10 ± 0.05. The fermented yogurt was added into pudding base (yogurt inoculation), or LA-5 was directly inoculated (direct inoculation). During the storage of the puddings at 4±1oC for 4 wks, the quality characteristics (pH, acidity, texture) and the probiotics viability of pudding were measured and compared. As a result, the puddings with yogurt inoculation showed significantly lower pH and higher acidity than those with direct inoculation. In texture properties, including hardness, chewiness, and gumminess, the addition of rice powder increased those in milk pudding more but the addition of inulin in the milk-soymilk pudding more. The addition of rice powder increased probiotics viability more than inulin in milk and milk-soymilk puddings. Therefore, adding rice and/or inulin can potentially improve the probiotics viability and quality characteristics of pudding.