Phenotypic Assessment of Probiotic and Bacteriocinogenic Efficacy of Indigenous LAB Strains from Human Breast Milk

Breast milk is the combination of bioactive compounds and microflora that promote newborn’s proper growth, gut flora, and immunity. Thus, it is always considered the perfect food for newborns. Amongst their bioactives, probiotic communities—especially lactic acid bacteria (LAB)—are characterized from breast milk over the first month of parturition. In this study, seven LAB were characterized phenotypically and genotypically as Levilactobacillus brevis BDUMBT08 ({"type":"entrez-nucleotide","attrs":{"text":"MT673657","term_id":"1863149999"}}MT673657), L. gastricus BDUMBT09 ({"type":"entrez-nucleotide","attrs":{"text":"MT774596","term_id":"1873222494"}}MT774596), L. paracasei BDUMBT10 ({"type":"entrez-nucleotide","attrs":{"text":"MT775430","term_id":"1874262304"}}MT775430), L. brevis BDUMBT11 ({"type":"entrez-nucleotide","attrs":{"text":"MW785062","term_id":"2009762994"}}MW785062), L. casei BDUMBT12 ({"type":"entrez-nucleotide","attrs":{"text":"MW785063","term_id":"2009820773"}}MW785063), L. casei BDUMBT13 ({"type":"entrez-nucleotide","attrs":{"text":"MW785178","term_id":"2010296340"}}MW785178), and Brevibacillus brevis M2403 ({"type":"entrez-nucleotide","attrs":{"text":"MK371781","term_id":"1548854744"}}MK371781) from human breast milk. Their tolerance to lysozyme, acid, bile, gastric juice, pancreatic juice, and NaCl and potential for mucoadhesion, auto-aggregation, and co-aggregation with pathogens are of great prominence in forecasting their gut colonizing ability. They proved their safety aspects as they were negative for virulence determinants such as hemolysis and biofilm production. Antibiogram of LAB showed their sensitivity to more than 90% of the antibiotics tested. Amongst seven LAB, three isolates (L. brevis BDUMBT08 and BDUMBT11, and L. gatricus BDUMBT09) proved their bacteriocin producing propensity. Although the seven LAB isolates differed in their behavior, their substantial probiotic properties with safety could be taken as promising probiotics for further studies to prove their in vivo effects, such as health benefits, in humans.     

Evaluation of Probiotic Potential of Bacteriocinogenic Lactic Acid Bacteria Strains Isolated from Meat Products

In this study, the probiotic potential of five bacteriocin-producing lactic acid bacteria (LAB) strains, isolated from meat products, was investigated. They were presumptively identified as Lactococcus lactis subsp. cremoris CTC 204 and CTC 483, L. lactis subsp. hordinae CTC 484, and Lactobacillus plantarum CTC 368 and CTC 469 according to morphological, biochemical, and physiological characteristics. Analysis of genetic variability (random amplified polymorphic (RAPD)-PCR) and whole-cell proteins (SDS-PAGE) revealed similarity between Lactobacillus strains and variability among Lactococcus strains. The evaluation of the probiotic potential showed that the five LAB strains were tolerant to pH 2.0, and only strain CTC 469 was tolerant to the lowest concentration of the bile salts evaluated (0.1%). All strains showed survival or growth ability at 4, 25, and 37 °C, and tolerance at ??20 °C. Although strain CTC 204 in TSB Broth supplemented with MgSO₄ showed the highest intensity of biofilm production, this compound was produced by all of them. The safety assessment showed that no thermonuclease, hemolytic, or gelatinase activities were detected. All strains were resistant to erythromycin and sensitive to amoxicillin and phenoxymethylpenicillin; furthermore, CTC 204 was resistant to chloramphenicol, CTC 368 and CTC 469 to chloramphenicol and vancomycin, CTC 483 to tetracycline and vancomycin, and CTC 484 to clindamycin and chloramphenicol. The evaluated strains showed biogenic amine production; the lowest levels were produced by CTC 204 and CTC 368 strains. It was concluded that CTC 204 and CTC 368 strains have the greatest potential for becoming probiotics.     

Phenotypic and Genotypic Identification of the Most Acidifiers LAB Strains Isolated from Fermented Food

Biology Bulletin - Over 18 strains of lactic acid bacteria “LAB” isolated from different traditionally fermented products mainly sourdough and fermented vegetables, have been...     

酶类药物现状、问题及展望

随着生物制药的迅速发展,许多酶类药物应运而生,在治疗代谢疾病、心血管疾病、癌症等诸多疾病上发挥着越来越重要的作用。但是酶类药物也存在一些不足,如潜在的免疫原性、较短的体内半衰期,以及较差的组织靶向性,影响了酶类药物的疗效和应用。为克服这些缺点,人们已开发出多种技术,如通过糖基化、聚乙二醇修饰等分子工程技术提升酶蛋白药效,另一方面酶基因疗法也已成功用于多种酶缺陷疾病的治疗。基于酶类药物的迅速发展和广泛的应用前景,本文对酶类药物的现状进行较详细的阐述,并对酶类药物的优势、所存在的问题及未来发展趋势进行分析和评述。     

Bacteriocinogenic Potential of Bacillus amyloliquefaciens Isolated from Kimchi,a Traditional Korean Fermented Cabbage

Bacteriocin production is considered a favorable property for various beneficial cultures. In addition to their potential as biopreservatives, bacteriocins are also promising alternatives for the control of multidrug-resistant pathogens and the inhibition of some viruses and cancer cells. The objective of this study was to screen and characterize a bacteriocin-producing strain with the aim of its future application for control of Listeria monocytogenes, an important food-borne pathogen. A total of 22 potentially bacteriocinogenic strains active against L. monocytogenes ATCC15313 were isolated from locally produced kimchi through a three-level approach. Pure cultures were obtained according to good microbiological practices and differentiated through RAPD-PCR using the primers OPL01, OPL09, and OPL11. Altogether, 5 strains were selected for further study. Specific focus was given to strain ST05DL based on its specific inhibitory activity against L. monocytogenes ATCC15313, while not affecting different strains belonging to the genera Lactobacillus, Pediococcus, Leuconostoc, and Weissella, most of which are beneficial microorganisms. The strain ST05DL was identified as Bacillus amyloliquefaciens based on its sugar fermentation profile obtained through API50CHB analysis and 16S rRNA partial sequencing. The antimicrobial compound produced by B. amyloliquefaciens ST05DL was found to be sensitive to pepsin and α-chymotrypsin, evidence of its proteinaceous nature. The presence of skim milk, NaCl, Tween 80, glycerol, and SDS did not affect the antimicrobial activity. The addition of 20% cell-free supernatant (CFS) obtained from a 24-h culture of B. amyloliquefaciens ST05DL to an exponentially growing culture of L. monocytogenes ATCC15313 successfully inhibited the test microorganisms during the monitored 10-h incubation. Optimal bacteriocin production by B. amyloliquefaciens ST05DL was observed during the stationary phase at 12 h (800 AU/mL) and remained stable for the next 15 h. The ratio between live and dead cells during this period was 74.37% and 25.66%, respectively, as determined by flow cytometry. The presence of the virulence genes hblA, hblB, hblC, nheA, nheB, and nheC was not detected in the total DNA of B. amyloliquefaciens ST05DL, and the strain was resistant only to ampicillin out of 10 tested antibiotics. Future evaluation of expressed bacteriocin/s by B. amyloliquefaciens ST05DL (amino acid sequence, molecular mass, cytotoxicity, detailed mode of action, etc.), will be the next step in the characterization and its potential application as biopreservative and/or pharmaceutical product.

     

Investigation into the Potential of Bacteriocinogenic Lactobacillus plantarum BFE 5092 for Biopreservation of Raw Turkey Meat

The bacteriocin-producing Lactobacillus plantarum BFE 5092 was assessed for its potential as a protective culture in the biopreservation of aerobically stored turkey meat. This strain produces three bacteriocins, i.e. plantaricins EF, JK and N. The absolute expression of Lactobacillus plantarum BFE 5092 16S rRNA housekeeping gene, as well as l-ldh, plnEF and plnG genes as determined by quantitative, real-time-PCR, revealed that these genes were expressed to similar levels when the strain was grown at 8 and 30 °C in MRS broth. On turkey meat, Lactobacillus plantarum BFE 5092 did not grow but survived, as indicated by similar viable cell numbers during a 9-day storage period at 8 °C. When inoculated at 1 × 10⁷ CFU/g on the turkey meat and subsequently stored at 10 °C, the culture did again not show good growth. Lactobacillus plantarum BFE 5092 could not inhibit the growth of naturally occurring listeriae or Gram-negative bacteria on the turkey meat at 10 °C, or that of Listeria monocytogenes when it was co-inoculated at a level of 1 × 10⁵ CFU/g. Gene expression analyses showed that the bacteriocin genes were expressed on turkey meat stored at 10 °C. Moreover, the investigation into the absolute expression of the three plantaricin genes of Lactobacillus plantarum BFE 5092 in co-culture with Listeria monocytogenes on turkey meat by qRT-PCR showed that the plantaricin genes were indeed expressed during the low-temperature storage condition. The Lactobacillus plantarum BFE 5092 strain overall could not effectively inhibit L. monocytogenes and therefore it would not make a suitable protective culture for biopreservation of turkey meat stored aerobically at low temperature.     

野生鸡枞菌种长期保存方法比较

野生鸡枞菌种质资源的有效保存是对野生鸡枞加以保护和利用的前提。以自行分离的5个野生鸡枞菌株作为研究对象,采用蒸馏水保藏法和-80°C冻结保藏法对野生鸡枞菌种长期保存的方法进行了实验研究,蒸馏水法分别保存于室温和4°C,-80°C冻结保藏同时采用程控降温法和泡沫盒降温法,保存20个月后对4种不同方法保存的5个菌株的保存效果进行比较。实验结果表明:蒸馏水室温保存法菌种存活率为100%,萌发期较短,为4-10 d,是一种简便、实用、有效而成本低廉的长期保存方法;-80°C冻结保藏法的存活率为56%-76%,萌发期7-16 d,泡沫盒降温法可以很好地控制降温速度,是一种简便有效的控温方法。     

Biomaterials for High‐Energy Lithium‐Based Batteries: Strategies,Challenges, and Perspectives

Developing high‐performance batteries through applying renewable resources is of great significance for meeting ever‐growing energy demands and sustainability requirements. Biomaterials have overwhelming advantages in material abundance, environmental benignity, low cost, and more importantly, multifunctionalities from structural and compositional diversity. Therefore, significant and fruitful research on exploiting various natural biomaterials (e.g., soy protein, chitosan, cellulose, fungus, etc.) for boosting high‐energy lithium‐based batteries by means of making or modifying critical battery components (e.g., electrode, electrolyte, and separator) are reported. In this review, the recent advances and main strategies for adopting biomaterials in electrode, electrolyte, and separator engineering for high‐energy lithium‐based batteries are comprehensively summarized. The contributions of biomaterials to stabilizing electrodes, capturing electrochemical intermediates, and protecting lithium metal anodes/enhancing battery safety are specifically emphasized. Furthermore, advantages and challenges of various strategies for fabricating battery materials via biomaterials are described. Finally, future perspectives and possible solutions for further development of biomaterials for high‐energy lithium‐based batteries are proposed.     

Isolation and Characterization of Pediocin NV 5 Producing <Emphasis Type="Italic">Pediococcus acidilactici</Emphasis> LAB 5 from Vacuum-Packed Fermented Meat Product

A potentially novel antimicrobial compound producing Pediococcus acidilactici LAB 5 was isolated from vacuum-packed fermented meat product. This compound was found active against some species of Enterococcus, Leuconostoc, Staphylococcus and Listeria, many of which are associated with food spoilage and food related health hazards. The strain was found to be a paired cocci which can utilize a broad range of carbohydrates and produce acid identical to the P. acidilactici and P. pentoseus. Since the antimicrobial agent was sensitive to proteolytic enzymes but quite resistant to heat, it was identified as a bacteriocin and was designated as Pediocin NV 5. The molecular weight of the bacteriocin was 10.3 kDa and the bacterium possessed a 5 kbp plasmid responsible for bacteriocin production and also for vancomycin resistance phenotype.     

Anaerobic Processes as the Core Technology for Sustainable Domestic Wastewater Treatment: Consolidated Applications, New Trends, Perspectives, and Challenges

Anaerobic digesters have been responsible for the removal of large fraction of organic matter (mineralization of waste sludge) in conventional aerobic sewage treatment plants since the early years of domestic sewage treatment (DST). Attention on the anaerobic technology for improving the sustainability of sewage treatment has been paid mainly after the energy crisis in the 1970s. The successful use of anaerobic reactors (especially up-flow anaerobic sludge blanket (UASB) reactors) for the treatment of raw domestic sewage in tropical and sub-tropical regions (where ambient temperatures are not restrictive for anaerobic digestion) opened the opportunity to substitute the aerobic processes for the anaerobic technology in removal of the influent organic matter. Despite the success, effluents from anaerobic reactors treating domestic sewage require post-treatment in order to achieve the emission standards prevailing in most countries. Initially, the composition of this effluent rich in reduced compounds has required the adoption of post-treatment (mainly aerobic) systems able to remove the undesirable constituents. Currently, however, a wealth of information obtained on biological and physical-chemical processes related to the recovery or removal of nitrogen, phosphorus and sulfur compounds creates the opportunity for new treatment systems. The design of DST plant with the anaerobic reactor as core unit coupled to the pre- and post-treatment systems in order to promote the recovery of resources and the polishing of effluent quality can improve the sustainability of treatment systems. This paper presents a broader view on the possible applications of anaerobic treatment systems not only for organic matter removal but also for resources recovery aiming at the improvement of the sustainability of DST.