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Effect of Probiotics on Sperm Quality in the Adult Mouse. 益生菌对成年小鼠精子质量的影响
IF 4.4 2区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-10-23 DOI: 10.1007/s12602-024-10388-z
Ana Sanchez-Rodriguez, Ingrid I D Idrovo, Rocío Villafranca, Nerea Latorre, Juan Antonio Rielo, Ane Laburu, Sandra Nieto-Román, Daniel Heredia, Rubén González, Virginia García-Cañas, Diego Laxalde, Carolina Simó, David R Vieites, Eduardo R S Roldan

The administration of probiotics for the treatment of different diseases has gained interest in recent years. However, few studies have evaluated their effects on reproductive traits. The objective of this study was to examine the effect of two mixtures of probiotics, a commercial probiotic (Vivomixx®) and a mix of Lacticaseibacillus rhamnosus GG and Faecalibacterium duncaniae A2-165, on sperm quality in a mouse model. Adult male mice (8 months old) were used for two experimental and one control groups (n = 5 each). The probiotics or physiological serum (control) was administered orally, twice a week, during 5 weeks. Sperm were collected from the cauda epididymis, and their total number, motility, kinematics, morphology, and acrosome integrity were assessed in recently collected samples and after a 60-min in vitro incubation. Results showed a higher percentage of normal sperm in both experimental groups, with fewer head abnormalities than in the control. Differences were found among groups in the morphometry of sperm heads, being more elongated in mice treated with probiotics. Sperm from probiotic-treated mice showed similar total motility when compared to the controls, although the proportion of progressively moving sperm and their vigor of motility were lower. Sperm swimming descriptors were measured with a CASA system. Velocity parameters were similar among groups whereas linearity was higher in mice treated with the commercial probiotic. These results suggest that the administration of probiotics may increase the proportion of sperm with normal morphology and lead to modifications in sperm head shape that may enhance sperm swimming. Studies using a longer administration period would be useful in further characterizing the effect of these probiotic mixtures on sperm quality and fertilization capacity.

近年来,使用益生菌治疗不同疾病的研究越来越受到关注。然而,很少有研究评估其对繁殖性状的影响。本研究旨在考察两种益生菌混合物(一种商用益生菌(Vivomixx®)和一种鼠李糖乳杆菌 GG 与粪肠球菌 A2-165 的混合物)对小鼠精子质量的影响。成年雄性小鼠(8 个月大)分为两个实验组和一个对照组(每组 5 只)。实验组口服益生菌或生理血清(对照组),每周两次,为期 5 周。从附睾尾部收集精子,在最近收集的样本中和经过 60 分钟的体外培养后,对精子的总数、运动能力、运动学、形态学和顶体完整性进行评估。结果显示,与对照组相比,两个实验组的正常精子比例更高,头部畸形的精子更少。不同实验组的精子头部形态存在差异,使用益生菌的小鼠精子头部更长。与对照组相比,经益生菌处理的小鼠精子的总运动能力相似,但逐渐运动的精子比例和运动活力较低。精子游动描述符是用 CASA 系统测量的。各组之间的速度参数相似,而使用商业益生菌治疗的小鼠的线性度更高。这些结果表明,服用益生菌可能会增加具有正常形态的精子比例,并导致精子头部形状的改变,从而增强精子的游动能力。延长给药时间的研究将有助于进一步确定这些益生菌混合物对精子质量和受精能力的影响。
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引用次数: 0
Alteration of Cecal Microbiota by Antimicrobial Peptides Enhances the Rational and Efficient Utilization of Nutrients in Holstein Bulls. 抗菌肽对盲肠微生物群的改变可提高荷斯坦公牛对营养物质的合理有效利用。
IF 4.4 2区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-10-23 DOI: 10.1007/s12602-024-10379-0
Jinping Shi, Yu Lei, Zemin Li, Li Jia, Pengjia He, Qiang Cheng, Zhao Zhang, Zhaomin Lei

We previously observed that supplementation with antimicrobial peptides facilitated the average daily weight gain, net meat, and carcass weights of Holstein bulls. To expand our knowledge of the possible impact of antimicrobial peptides on cecum microbiota, further investigations were conducted. In this study, 18 castrated Holstein bulls with insignificant weight differences and 10 months of age were split randomly into two groups. The control group (CK) was fed a basic diet, whereas the antimicrobial peptide group (AP) was supplemented with 8 g of antimicrobial peptides for 270 days. After slaughter, metagenomic and metabolomic sequencing analyses were performed on the cecum contents. The results showed significantly higher levels of amylase, cellulase, protease, and lipase in the CK than in the AP group (P ≤ 0.05). The levels of β-glucosidase and xylanase (P ≤ 0.05), and acetic and propionic acids (P ≤ 0.01), were considerably elevated in the AP than in the CK group. The metagenome showed variations between the two groups only at the bacterial level, and 3258 bacteria with differences were annotated. A total of 138 differential abundant genes (P < 0.05) were identified in the CAZyme map, with 65 genes more abundant in the cecum of the AP group and 48 genes more abundant in the cecum of the CK group. Metabolomic analysis identified 68 differentially expressed metabolites. Conjoint analysis of microorganisms and metabolites revealed that Lactobacillus had the greatest impact on metabolites in the AP group and Brumimicrobium in the CK group. The advantageous strains of the AP group Firmicutes bacterium CAG:110 exhibited a strong symbiotic relationship with urodeoxycholic acid and hyodeoxycholic acid. This study identified the classification characteristics, functions, metabolites, and interactions of cecal microbiota with metabolites that contribute to host growth performance. Antimicrobial peptides affect the cecal microorganisms, making the use of nutrients more efficient. The utilization of hemicellulose in the cecum of ruminants may contribute more than cellulose to their production performance.

我们以前曾观察到,补充抗菌肽有助于提高荷斯坦公牛的平均日增重、净肉和胴体重量。为了进一步了解抗菌肽对盲肠微生物群可能产生的影响,我们进行了进一步的研究。在这项研究中,18 头体重差异不明显、年龄为 10 个月的阉割荷斯坦公牛被随机分成两组。对照组(CK)饲喂基本饲料,而抗菌肽组(AP)则连续 270 天添加 8 克抗菌肽。屠宰后,对盲肠内容物进行了元基因组和代谢组测序分析。结果显示,CK 组的淀粉酶、纤维素酶、蛋白酶和脂肪酶水平明显高于 AP 组(P ≤ 0.05)。AP组的β-葡萄糖苷酶和木聚糖酶水平(P≤0.05)以及乙酸和丙酸水平(P≤0.01)均明显高于CK组。元基因组仅在细菌水平上显示了两组之间的差异,共注释了 3258 个存在差异的细菌。共有 138 个差异丰度基因(P
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引用次数: 0
Genomic Characterization and Probiotic Properties of Lactiplantibacillus pentosus Isolated from Fermented Rice. 从发酵大米中分离出的五联乳杆菌的基因组特征和益生特性
IF 4.4 2区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-10-21 DOI: 10.1007/s12602-024-10378-1
Athira Cheruvari, Rajagopal Kammara

The aim of the study was the preliminary genetic and phenotypic characterization of a potential probiotic strain of Lactiplantibacillus pentosus (strain krglsrbmofpi2) obtained from traditionally fermented rice. Genome sequencing revealed that the strain has a 3.7-Mb genome with a GC content of 46 and a total of 3192 protein-coding sequences. Using bioinformatic methods, we have successfully identified phage genes, plasmids, pathogenicity, antibiotic resistance and a variety of bacteriocins. Through comprehensive biochemical and biophysical analyses, we have gained valuable insights into its auto-aggregation, co-aggregation, antibiotic resistance, hydrophobicity, antioxidant activity and tolerance to simulated gastrointestinal conditions. The safety evaluation of the isolated L. pentosus was performed on the basis of its haemolytic activity. Our studies have shown that this strain has a strong antagonistic activity against the priority pathogens identified by the World Health Organization such as Vibrio cholerae, Clostridium perfringens, Salmonella enterica subsp. enterica ser. Typhi, Escherichia coli, Listeria monocytogenes and Staphylococcus aureus. It is essential to fully understand the genetic and functional properties of the L. pentosus strain before considering its use as a useful probiotic in the food industry.

本研究的目的是对从传统发酵大米中获得的潜在益生菌株戊糖乳杆菌(菌株 krglsrbmofpi2)进行初步遗传和表型鉴定。基因组测序显示,该菌株的基因组为 3.7 兆字节,GC 含量为 46,共有 3192 个蛋白质编码序列。利用生物信息学方法,我们成功鉴定了噬菌体基因、质粒、致病性、抗生素抗性和多种细菌素。通过全面的生物化学和生物物理分析,我们对其自动聚集、共聚集、抗生素耐药性、疏水性、抗氧化活性和对模拟胃肠道条件的耐受性有了宝贵的认识。根据其溶血活性,对分离出的五味子乳杆菌进行了安全性评估。我们的研究表明,该菌株对世界卫生组织确定的重点病原体具有很强的拮抗活性,如霍乱弧菌、产气荚膜梭菌、肠炎沙门氏菌亚种、伤寒杆菌、大肠埃希氏菌等。这些病原体包括霍乱弧菌、产气荚膜梭菌、肠炎沙门氏菌亚种、伤寒杆菌、大肠杆菌、李斯特菌和金黄色葡萄球菌。在考虑将五味子乳杆菌菌株用作食品工业中有用的益生菌之前,必须充分了解其遗传和功能特性。
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引用次数: 0
A Review on Bacteriocin Extraction Techniques from Lactic Acid Bacteria. 从乳酸菌中提取细菌素技术综述
IF 4.4 2区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-10-21 DOI: 10.1007/s12602-024-10384-3
Sharleen Livina Isaac, Amalia Mohd Hashim, Fadzlie Wong Faizal Wong, Muhamad Afiq Mohamed Akbar, Wan Nur Ismah Wan Ahmad Kamil

Lactic acid bacteria (LAB) are widely known for the production of secondary metabolites such as organic acids and other bioactive compounds such as bacteriocins. Finding a broad application in food and healthcare, bacteriocins have received increased attention due to their inherent antimicrobial properties. However, the extraction of bacteriocins is often plagued with low yields due to the complexity of the extraction processes and the diversity of bacteriocins themselves. Here, we review the current knowledge related to bacteriocin extraction on the different extraction techniques for isolating bacteriocins from LAB. The advantages and disadvantages of each technique will also be critically appraised, taking into account factors such as extraction efficiency, scalability and cost-effectiveness. This review aims to guide researchers and professionals in selecting the most suitable approach for bacteriocin extraction from LAB by illuminating the respective advantages and limitations of various extraction techniques.

乳酸菌(LAB)因生产有机酸等次级代谢产物和细菌素等其他生物活性化合物而广为人知。由于其固有的抗菌特性,细菌素在食品和医疗保健领域有着广泛的应用,受到越来越多的关注。然而,由于提取过程的复杂性和细菌素本身的多样性,细菌素的提取往往存在产量低的问题。在此,我们回顾了目前与细菌素提取相关的知识,即从 LAB 中分离细菌素的不同提取技术。考虑到提取效率、可扩展性和成本效益等因素,我们还将对每种技术的优缺点进行严格评估。本综述旨在通过阐明各种提取技术各自的优势和局限性,指导研究人员和专业人员选择最适合的方法从 LAB 中提取细菌素。
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引用次数: 0
Heyndrickxia coagulans LMG S-24828 Is a Safe Probiotic Strain Capable of Germinating in the Human Gut. Heyndrickxia coagulans LMG S-24828 是一种能够在人体肠道中发芽的安全益生菌株。
IF 4.4 2区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-10-21 DOI: 10.1007/s12602-024-10383-4
Robin Duncan, Giacomo Mantegazza, Giorgio Gargari, Elena Pierallini, Rosario Russo, Simone Guglielmetti

Ensuring the viability and efficacy of probiotic microorganisms during manufacturing and gastrointestinal transit remains challenging, particularly for sensitive strains such as certain lactic acid bacteria and bifidobacteria. This has led to increased interest in spore-forming bacteria, such as Heyndrickxia coagulans (formerly Bacillus coagulans), which can endure environmental stresses through their endospore forms. This study presents a comprehensive analysis of the probiotic potential of strain LMG S-24828, originally isolated from healthy human feces. The genomic analysis confirmed the strain's taxonomic placement within the species H. coagulans and revealed no extrachromosomal plasmid DNA, suggesting genetic stability. Safety assessments demonstrated that LMG S-24828 does not produce D-lactate, deconjugate bile salts, or exhibit hemolytic activity, and it lacks transmissible antibiotic resistances. Phenotypic tests showed the strain's metabolic versatility, including its ability to hydrolyze complex carbohydrates and adhere to intestinal epithelial cells. Moreover, LMG S-24828 exhibited robust survival and germination during in vitro and in vivo gastrointestinal simulations, with evidence of significant spore germination in the human gut. These findings suggest that H. coagulans LMG S-24828 possesses several advantageous traits for probiotic applications, warranting further clinical evaluation to confirm its health benefits.

在生产和胃肠道转运过程中确保益生菌微生物的活力和功效仍具有挑战性,尤其是对某些乳酸菌和双歧杆菌等敏感菌株而言。因此,人们对凝结芽孢杆菌(Heyndrickxia coagulans,前身为凝结芽孢杆菌)等孢子形成菌越来越感兴趣,因为它们可以通过内生孢子形式承受环境压力。本研究对最初从健康人粪便中分离出来的菌株 LMG S-24828 的益生潜力进行了全面分析。基因组分析确认了该菌株在分类学上属于凝结球菌属(H. coagulans),并且没有发现染色体外的质粒 DNA,这表明该菌株具有遗传稳定性。安全性评估表明,LMG S-24828 不产生 D-乳酸,不分解胆盐,也不表现出溶血活性,而且缺乏可传播的抗生素耐药性。表型测试表明,该菌株具有多功能代谢能力,包括水解复杂碳水化合物和粘附肠上皮细胞的能力。此外,LMG S-24828 在体外和体内胃肠道模拟试验中表现出强大的存活和发芽能力,并有证据表明其孢子在人体肠道中大量发芽。这些研究结果表明,凝结球菌 LMG S-24828 具有益生菌应用的若干优势特性,值得进一步进行临床评估,以确认其对健康的益处。
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引用次数: 0
Unveiling the Potential of Protein-Based Sustainable Antibacterial Materials. 揭示基于蛋白质的可持续抗菌材料的潜力。
IF 4.4 2区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-10-18 DOI: 10.1007/s12602-024-10381-6
Iqra Ghouri, Muslum Demir, Shahid Ali Khan, Muhammad Adil Mansoor, Mudassir Iqbal

The surge in bacterial growth and the escalating resistance against a multitude of antibiotic drugs have burgeoned into an alarming global threat, necessitating urgent and innovative interventions. In response to this peril, scientists have embarked on the development of advanced biocompatible antibacterial materials, aiming to counteract not only bacterial infections but also the pervasive issue of food spoilage resulting from microbial proliferation. Protein-based biopolymers and their meticulously engineered composites are at the forefront of this endeavor. Their potential in combating this severe global concern presents an approach that intersects the domains of biomedicine and environmental science. The present review article delves into the intricate extraction processes employed to derive various proteins from their natural sources, unraveling the complex biochemical pathways that underpin their antibacterial properties. Expanding on the foundational knowledge, the review also provides a comprehensive synthesis of functionalized proteins modified to enhance their antibacterial efficacy, unveiling a realm of possibilities for tailoring solutions to specific biomedical and environmental applications. The present review navigates through their antibacterial applications; from wound dressings to packaging materials with inherent antibacterial properties, the potential applications underscore the versatility and adaptability of these materials. Moreover, this comprehensive review serves as a valuable roadmap, guiding future research endeavors in reshaping the landscape of natural antibacterial materials on a global scale.

细菌数量激增,对多种抗生素药物的耐药性不断升级,已成为一个令人担忧的全球性威胁,需要采取紧急和创新的干预措施。为了应对这一危险,科学家们已着手开发先进的生物相容性抗菌材料,目的不仅是为了应对细菌感染,还为了应对微生物繁殖导致的食品腐败这一普遍问题。以蛋白质为基础的生物聚合物及其精心设计的复合材料正处于这一领域的前沿。它们在解决这一全球关注的严重问题方面的潜力提供了一种生物医学和环境科学领域交叉的方法。本综述文章深入探讨了从天然来源中提取各种蛋白质所采用的复杂提取工艺,揭示了支撑其抗菌特性的复杂生化途径。在基础知识的基础上,这篇综述还对功能化蛋白质进行了全面综合,对其进行修饰以增强其抗菌功效,从而为特定的生物医学和环境应用提供量身定制的解决方案。本综述介绍了这些材料的抗菌应用;从伤口敷料到具有固有抗菌特性的包装材料,其潜在应用凸显了这些材料的多功能性和适应性。此外,本综述还是一份宝贵的路线图,可指导未来的研究工作,在全球范围内重塑天然抗菌材料的面貌。
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引用次数: 0
Shaping the Future of Functional Foods: Using 3D Printing for the Encapsulation and Development of New Probiotic Foods. 塑造功能食品的未来:利用三维打印技术封装和开发新型益生菌食品。
IF 4.4 2区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-10-18 DOI: 10.1007/s12602-024-10382-5
Josemar Gonçalves de Oliveira Filho, Larissa Graziele Rauber Duarte, Diego Oliveira Bonfim, Mateus Kawata Salgaço, Luiz Henrique Caparelli Mattoso, Mariana Buranelo Egea

Consumers have been demanding foods that, besides providing nutrition, bring some health benefits, known as functional foods. The insertion of probiotics in foods is a strategy for developing functional foods. Still, it has been a challenge because these matrices have different pHs and undergo different process temperatures and times that can reduce the viability of these microorganisms. In this sense, encapsulation using 3D printing emerges to protect probiotic microorganisms and ensure that they reach the intestine viable and carry out the expected beneficial action. Thus, this review evaluates the current advancements in 3D printing to encapsulate and develop novel probiotic foods. Research has shown that 3D printing effectively encapsulates probiotic microorganisms, preserving their viability throughout the gastrointestinal tract. Studies have proven the effectiveness of 3D printing encapsulation in protecting probiotics during processing, storage, and digestion. Innovative formulations for 3D bioprinted products with probiotics, such as food structures based on cereals, mashed potatoes, and cream, have been developed. Producing products with shelf life and combining applications of phytochemicals and probiotics aims to improve personalized nutrition, textural characteristics, and sensory attributes of the foods produced by this emerging approach. Therefore, 3D printing of foods with probiotics has the potential to create new products that meet this demand.

消费者一直要求食品除了提供营养外,还能带来一些健康益处,这就是所谓的功能性食品。在食品中添加益生菌是开发功能食品的一种策略。然而,这仍然是一项挑战,因为这些基质具有不同的 pH 值,并经历不同的加工温度和时间,这些都会降低这些微生物的活力。从这个意义上说,利用三维打印技术进行封装可以保护益生菌微生物,并确保它们能在肠道中存活并发挥预期的有益作用。因此,本综述评估了目前在三维打印封装和开发新型益生菌食品方面取得的进展。研究表明,三维打印技术能有效封装益生菌微生物,在整个胃肠道中保持其活力。研究证明,3D 打印封装技术可在加工、储存和消化过程中有效保护益生菌。目前已开发出含有益生菌的 3D 生物打印产品的创新配方,如基于谷物、土豆泥和奶油的食品结构。生产具有保质期的产品,并结合植物化学物质和益生菌的应用,旨在改善这种新兴方法生产的食品的个性化营养、质地特征和感官属性。因此,含有益生菌的 3D 打印食品有可能创造出满足这一需求的新产品。
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引用次数: 0
Complete Genome Sequence and Probiotic Characterization of Lactobacillus delbrueckii subsp. Indicus DC-3 Isolated from Traditional Indigenous Fermented Milk. 从传统土著发酵乳中分离的德尔布鲁贝克乳杆菌亚种(Lactobacillus delbrueckii subsp.
IF 4.4 2区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-10-17 DOI: 10.1007/s12602-024-10385-2
Deepti N Chaudhari, Jayesh J Ahire, Anupama N Devkatte, Amit A Kulthe

In this study, Lactobacillus delbrueckii subsp. indicus DC-3 was isolated from Indian traditional indigenous fermented milk Dahi and identified using whole genome sequencing. The safety of the strain was evaluated using genetic and phenotypic analyses, such as the presence of virulence factors, mobile and insertion elements, plasmids, antibiotic resistance, etc. Besides this, the strain was comprehensively investigated for in vitro probiotic traits, biofilm formation, antibacterials, and exopolysaccharide (EPS) production. In results, the strain showed a single circular chromosome (3,145,837 bp) with a GC content of 56.73%, a higher number of accessory and unique genes, an open pan-genome, and the absence of mobile and insertion elements, plasmids, virulence, and transmissible antibiotic resistance genes. The strain was capable of surviving in gastric juice (83% viability at 3 h) and intestinal juice (71% viability at 6 h) and showed 42.5% autoaggregation, adhesion to mucin, 8.7% adhesion to xylene, and 8.3% adhesion to Caco-2 cells. The γ-hemolytic nature, usual antibiotic susceptibility profile, and negative results for mucin and gelatin degradation ensure the safety of the strain. The strain produced 10.5 g/L of D-lactic acid and hydrogen peroxide, capable of inhibiting and co-aggregating Escherichia coli MTCC 1687, Proteus mirabilis MTCC 425, and Candida albicans ATCC 14,053. In addition, the strain showed 90 mg/L EPS (48 h) and biofilm formation. In conclusion, this study demonstrates that L. delbrueckii subsp. indicus DC-3 is unique and different than previously reported L. delbrueckii subsp. indicus strains and is a safe potential probiotic candidate.

本研究从印度传统本土发酵牛奶 Dahi 中分离出了 Lactobacillus delbrueckii subsp.利用遗传和表型分析评估了该菌株的安全性,如是否存在毒力因子、移动和插入元件、质粒、抗生素耐药性等。此外,还对该菌株的体外益生特性、生物膜形成、抗菌和外多糖(EPS)产生进行了全面研究。结果显示,该菌株具有单环染色体(3,145,837 bp),GC 含量为 56.73%,有较多的附属基因和独特基因,泛基因组开放,不存在移动和插入元件、质粒、毒力基因和可传播的抗生素耐药基因。该菌株能在胃液(3 小时后存活率为 83%)和肠液(6 小时后存活率为 71%)中存活,并显示出 42.5% 的自身聚集性、对粘蛋白的粘附性、对二甲苯的 8.7% 粘附性和对 Caco-2 细胞的 8.3% 粘附性。该菌株的γ-溶血性、通常的抗生素敏感性以及粘蛋白和明胶降解的阴性结果确保了其安全性。该菌株能产生 10.5 克/升的 D-乳酸和过氧化氢,能够抑制和共聚大肠杆菌 MTCC 1687、变形杆菌 MTCC 425 和白色念珠菌 ATCC 14 053。此外,该菌株还表现出 90 毫克/升的 EPS(48 小时)和生物膜形成。总之,本研究证明了 L. delbrueckii 亚种 indicus DC-3 与之前报道的 L. delbrueckii 亚种 indicus 菌株相比具有独特性和差异性,是一种安全的潜在候选益生菌。
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引用次数: 0
Characterization of a Peptidoglycan-Degrading Protein: Biochemical and Antimicrobial Characteristics, Antibiotic Synergism, and Delivery System Innovation. 肽聚糖降解蛋白的特征:一种肽聚糖降解蛋白的特性:生化和抗菌特性、抗生素协同作用和给药系统创新。
IF 4.4 2区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-10-16 DOI: 10.1007/s12602-024-10374-5
Viviane C Oliveira, Nathália G Rosa-Garzon, Amanda C S D Rocha, Rachel M Monteiro, Yasmim G Gonçalves, Marcelo Kravicz, Maria A Juliano, Luiz Juliano, Priscyla D Marcato, Claudia H Silva-Lovato, Hamilton Cabral, Evandro Watanabe

The global health threat posed by antibiotic resistance has led to new research involving bacteriophage-encoded enzymes. This study characterized a new peptidoglycan-degrading protein and evaluated its synergism with colistin and its antimicrobial efficacy when conjugated with polycationic-polymer nanoparticles. The gene that codes for endolysin in the vB_PaeM_USP2, a Pseudomonas aeruginosa bacteriophage, was cloned and expressed in Escherichia coli. The recombinant endolysin (rEnd2) was purified and its biochemical properties were determined using peptidoglycan substrate. The enzymatic activity was measured through peptidoglycan layer degradation and a decrease in turbidity of permeabilized Gram-negative bacteria. The antimicrobial activity of rEnd2, alone and in combination with colistin, was evaluated by checkerboard assay. The antibacterial activity of the cationic lipid oleylamine (OAM) conjugated with rEnd2 (OAM-rEnd2) was evaluated by time killing assay. The rEnd2 is structurally analogue with other endolysins and showed muramidase activity. The rEnd2 maintained higher activity between pH 6.0 to 7.5, had maximum activity at 35 °C, and was not affected by chaotropic and reducing reagents. It was sensitive to an increase in surfactant concentration, being inactivated by sodium dodecyl sulfate and cetyltrimethylammonium bromide. Ions exhibited neither a positive nor a negative effect on enzyme activity. The rEnd2 showed clear muralytic activity and decreased turbidity of permeabilized Gram-negative bacteria. However, it did not control bacterial growth despite the combination with an antibiotic and its complexation with polycation (OAM-rEnd2 nanoparticle conjugate). The rEnd2 did not show clear antimicrobial activity suggesting further optimization of conditions for its activity or engineering and modification.

抗生素耐药性对全球健康的威胁促使人们对噬菌体编码酶进行新的研究。这项研究对一种新的肽聚糖降解蛋白进行了鉴定,并评估了它与可乐定的协同作用以及与多阳离子聚合物纳米颗粒共轭后的抗菌效果。在大肠杆菌中克隆并表达了铜绿假单胞菌噬菌体 vB_PaeM_USP2 中编码内溶素的基因。纯化了重组内溶菌素(rEnd2),并使用肽聚糖底物测定了其生化性质。酶活性是通过肽聚糖层降解和渗透革兰氏阴性菌浊度降低来测定的。通过棋盘试验评估了 rEnd2 单独或与可乐定联合使用时的抗菌活性。阳离子脂质油胺(OAM)与 rEnd2 共轭(OAM-rEnd2)的抗菌活性通过时间杀伤试验进行了评估。rEnd2 在结构上与其他内溶酶类似,具有莽草酶活性。rEnd2 在 pH 值为 6.0 至 7.5 之间保持较高的活性,在 35 °C 时活性最高,并且不受混沌试剂和还原试剂的影响。它对表面活性剂浓度的增加很敏感,十二烷基硫酸钠和十六烷基三甲基溴化铵会使其失活。离子对酶活性既无积极影响,也无消极影响。rEnd2 具有明显的壁化活性,可降低渗透革兰氏阴性细菌的浑浊度。然而,尽管它与抗生素结合并与多阳离子复合物(OAM-rEnd2 纳米粒子共轭物)复配,却不能控制细菌的生长。rEnd2 没有显示出明显的抗菌活性,这表明需要进一步优化其活性条件或对其进行工程和改性。
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引用次数: 0
A Mixture of Four Probiotic Strains (Probiatop®) Mitigates Food Allergy to Ovalbumin in Mice. 四种益生菌株的混合物(Probiatop®)能减轻小鼠对卵清蛋白的食物过敏。
IF 4.4 2区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-10-15 DOI: 10.1007/s12602-024-10386-1
Vivian C Miranda, Ramon O Souza, Bruno Gallotti, Marcos F A de Oliveira, Ana Maria C Faria, Jacques R Nicoli, Enio Ferreira, Denise C C Machado, Flaviano S Martins

Food allergy is a pathological condition caused by a disruption of oral tolerance. This condition leads to a pro-inflammatory environment that culminates in symptoms that varies from abdominal pain to anaphylaxis and death. The gold standard treatment consists of removing the food that triggers the allergy from diet. However, this conduct can cause nutritional impairment and social restrictions. Therefore, the need for new treatment strategies is notorious. In this context, probiotics are investigated due to their immunomodulatory mechanisms. Therefore, the objective of the present work is to investigate the probiotic potential of a mixture of four probiotic strains (Probiatop®) in an in vivo model of food allergy to ovalbumin (OVA). Our results demonstrated that oral administration of Probiatop® attenuated weight loss and diminished significantly anti-OVA IgE and IgG1 levels. Furthermore, it mitigated proximal jejunum injury, neutrophil recruitment and local IL17 levels. In addition, the probiotic mixture modulated positively the gut microbiota composition by decreasing the levels and frequency of Staphylococcus and yeast. In summary, our data suggest that Probiatop® has the potential to alleviate important symptoms of IgE-mediated food allergy, suggesting its probiotic potential as an adjuvant in the treatment of ovalbumin food allergy.

食物过敏是一种由口腔耐受性紊乱引起的病理状态。这种情况会导致一种促炎症环境,最终出现从腹痛到过敏性休克和死亡的各种症状。金标准疗法包括从饮食中去除引发过敏的食物。然而,这种做法会造成营养障碍和社交限制。因此,对新治疗策略的需求是众所周知的。在这种情况下,益生菌因其免疫调节机制而受到研究。因此,本研究的目的是调查四种益生菌株混合物(Probiatop®)在体内卵清蛋白(OVA)食物过敏模型中的益生潜力。我们的研究结果表明,口服 Probiatop® 可减轻体重下降,并显著降低抗 OVA IgE 和 IgG1 水平。此外,它还减轻了近端空肠损伤、中性粒细胞募集和局部 IL17 水平。此外,益生菌混合物通过降低葡萄球菌和酵母菌的水平和频率,积极调节了肠道微生物群的组成。总之,我们的数据表明,Probiatop® 有可能减轻 IgE 介导的食物过敏的重要症状,这表明其益生菌有可能成为治疗卵清蛋白食物过敏的辅助剂。
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引用次数: 0
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Probiotics and Antimicrobial Proteins
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