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Fulltext A Review on the Biotechnological Applications of the Operational Group Bacillus amyloliquefaciens

Ngalimat MS, Yahaya RSR, Baharudin MMA, Yaminudin SM, Karim M, Ahmad SA, et al.

Microorganisms, 2021 Mar 17;9(3).
PMID: 33802666 DOI: 10.3390/microorganisms9030614

Bacteria under the operational group Bacillus amyloliquefaciens (OGBa) are all Gram-positive, endospore-forming, and rod-shaped. Taxonomically, the OGBa belongs to the Bacillus subtilis species complex, family Bacillaceae, class Bacilli, and phylum Firmicutes. To date, the OGBa comprises four bacterial species: Bacillus amyloliquefaciens, Bacillus siamensis, Bacillus velezensis and Bacillus nakamurai. They are widely distributed in various niches including soil, plants, food, and water. A resurgence in genome mining has caused an increased focus on the biotechnological applications of bacterial species belonging to the OGBa. The members of OGBa are known as plant growth-promoting bacteria (PGPB) due to their abilities to fix nitrogen, solubilize phosphate, and produce siderophore and phytohormones, as well as antimicrobial compounds. Moreover, they are also reported to produce various enzymes including α-amylase, protease, lipase, cellulase, xylanase, pectinase, aminotransferase, barnase, peroxidase, and laccase. Antimicrobial compounds that able to inhibit the growth of pathogens including non-ribosomal peptides and polyketides are also produced by these bacteria. Within the OGBa, various B. velezensis strains are promising for use as probiotics for animals and fishes. Genome mining has revealed the potential applications of members of OGBa for removing organophosphorus (OPs) pesticides. Thus, this review focused on the applicability of members of OGBa as plant growth promoters, biocontrol agents, probiotics, bioremediation agents, as well as producers of commercial enzymes and antibiotics. Here, the bioformulations and commercial products available based on these bacteria are also highlighted. This review will better facilitate understandings of members of OGBa and their biotechnological applications.

Matched MeSH terms: Probiotics
Fulltext A Revolutionizing Approach to Autism Spectrum Disorder Using the Microbiome

Johnson D, Letchumanan V, Thurairajasingam S, Lee LH

Nutrients, 2020 Jul 03;12(7).
PMID: 32635373 DOI: 10.3390/nu12071983

The study of human microbiota and health has emerged as one of the ubiquitous research pursuits in recent decades which certainly warrants the attention of both researchers and clinicians. Many health conditions have been linked to the gut microbiota which is the largest reservoir of microbes in the human body. Autism spectrum disorder (ASD) is one of the neurodevelopmental disorders which has been extensively explored in relation to gut microbiome. The utilization of microbial knowledge promises a more integrative perspective in understanding this disorder, albeit being an emerging field in research. More interestingly, oral and vaginal microbiomes, indicating possible maternal influence, have equally drawn the attention of researchers to study their potential roles in the etiopathology of ASD. Therefore, this review attempts to integrate the knowledge of microbiome and its significance in relation to ASD including the hypothetical aetiology of ASD and its commonly associated comorbidities. The microbiota-based interventions including diet, prebiotics, probiotics, antibiotics, and faecal microbial transplant (FMT) have also been explored in relation to ASD. Of these, diet and probiotics are seemingly promising breakthrough interventions in the context of ASD for lesser known side effects, feasibility and easier administration, although more studies are needed to ascertain the actual clinical efficacy of these interventions. The existing knowledge and research gaps call for a more expanded and resolute research efforts in establishing the relationship between autism and microbiomes.

Matched MeSH terms: Probiotics/therapeutic use
Fulltext A metagenomic comparison of clearwater, probiotic, and Rapid BFTTM on Pacific whiteleg shrimp, Litopenaeus vannamei cultures

Waiho K, Abd Razak MS, Abdul Rahman MZ, Zaid Z, Ikhwanuddin M, Fazhan H, et al.

PeerJ, 2023;11:e15758.
PMID: 37790619 DOI: 10.7717/peerj.15758

Biofloc technology improves water quality and promote the growth of beneficial bacteria community in shrimp culture. However, little is known about the bacteria community structure in both water and gut of cultured organisms. To address this, the current study characterised the metagenomes derived from water and shrimp intestine samples of novel Rapid BFTTM with probiotic and clearwater treatments using 16S V4 region and full length 16S sequencing. Bacteria diversity of water and intestine samples of Rapid BFTTM and probiotic treatments were similar. Based on the 16S V4 region, water samples of >20 μm biofloc had the highest abundance of amplicon sequence variant (ASV). However, based on full length 16S, no clear distinction in microbial diversity was observed between water samples and intestine samples. Proteobacteria was the most abundant taxon in all samples based on both 16S V4 and full length 16S sequences. Vibrio was among the highest genus based on 16S V4 region but only full length 16S was able to discern up to species level, with three Vibrios identified-V. harveyi, V. parahaemolyticus and V. vulnificus. Vibrio harveyi being the most abundant species in all treatments. Among water samples, biofloc water samples had the lowest abundance of all three Vibrios, with V. vulnificus was present only in bioflocs of <20 μm. Predicted functional profiles of treatments support the beneficial impacts of probiotic and biofloc inclusion into shrimp culture system. This study highlights the potential displacement of opportunistic pathogens by the usage of biofloc technology (Rapid BFTTM) in shrimp culture.

Matched MeSH terms: Probiotics*
Fulltext A randomized double-blind placebo-controlled trial of probiotics in post-surgical colorectal cancer

Zaharuddin L, Mokhtar NM, Muhammad Nawawi KN, Raja Ali RA

BMC Gastroenterol, 2019 Jul 24;19(1):131.
PMID: 31340751 DOI: 10.1186/s12876-019-1047-4

BACKGROUND: Our study aimed to determine the effect of probiotic consumption containing six viable microorganisms of 30 × 1010 cfu Lactobacillus and Bifidobacteria strains for six months on clinical outcomes and inflammatory cytokines (TNF-α, IFN-γ, IL-6, IL-10, IL-12, IL-17A, IL-17C and IL-22) in patients with colorectal cancer.
METHODS: Fifty-two patients with colorectal cancer were randomized at four weeks after surgery to receive either a placebo (n = 25) or 30 billion colony-forming unit (CFU) of a mixture of six viable strains including 107 mg of Lactobacillus acidophilus BCMC® 12,130, Lactobacillus lactis BCMC® 12,451, Lactobacillus casei subsp BCMC® 12,313, Bifidobacterium longum BCMC® 02120, Bifidobacterium bifidum BCMC® 02290 and Bifidobacterium infantis BCMC® 02129 (n = 27). Patients were instructed to take the product orally twice daily for six months. Infection status, diarrhea or hospital admission were recorded throughout the study. Blood was taken pre- and post-intervention to measure TNF-α, IFN-γ, IL-6, IL-10, IL-12, IL-17A, IL-17C and IL-22 using ELISA multiplex kit.
RESULTS: The majority of cases (~ 70%) were in Duke's C colorectal cancer for both groups. No surgical infection occurred and no antibiotics were required. Chemotherapy induced diarrhea was observed in both groups. Significant reduction in the level of pro-inflammatory cytokine, TNF-α, IL-6, IL-10, IL-12, IL-17A, IL-17C and IL-22 were observed in CRC patients who received probiotics as compared to pre-treatment level (P

Matched MeSH terms: Probiotics
Fulltext A randomized, double-blind, placebo-controlled, clinical trial on probiotic soy milk and soy milk: effects on epigenetics and oxidative stress in patients with type II diabetes

Hariri M, Salehi R, Feizi A, Mirlohi M, Ghiasvand R, Habibi N

Genes Nutr, 2015 Nov;10(6):52.
PMID: 26577825 DOI: 10.1007/s12263-015-0503-1

This clinical trial aimed to discover the effects of probiotic soy milk and soy milk on MLH1 and MSH2 promoter methylation, and oxidative stress among type II diabetic patients. Forty patients with type II diabetes mellitus aged 35-68 years were assigned to two groups in this randomized, double-blind, controlled clinical trial. Patients in the intervention group consumed 200 ml/day of probiotic soy milk containing Lactobacillus plantarum A7, while those in the control group consumed 200 ml/d of conventional soy milk for 8 weeks. Fasting blood samples, anthropometric measurements, and 24-h dietary recalls were collected at the baseline and at the end of the study, respectively. Probiotic soy milk significantly decreased promoter methylation in proximal and distal MLH1 promoter region (P 0.05). The consumption of probiotic soy milk improved antioxidant status in type II diabetic patients and may decrease promoter methylation among these patients, indicating that probiotic soy milk is a promising agent for diabetes management.

Matched MeSH terms: Probiotics
Fulltext A review on Lactococcus lactis: from food to factory

Song AA, In LLA, Lim SHE, Rahim RA

Microb Cell Fact, 2017 04 04;16(1):55.
PMID: 28376880 DOI: 10.1186/s12934-017-0669-x

Lactococcus lactis has progressed a long way since its discovery and initial use in dairy product fermentation, to its present biotechnological applications in genetic engineering for the production of various recombinant proteins and metabolites that transcends the heterologous species barrier. Key desirable features of this gram-positive lactic acid non-colonizing gut bacteria include its generally recognized as safe (GRAS) status, probiotic properties, the absence of inclusion bodies and endotoxins, surface display and extracellular secretion technology, and a diverse selection of cloning and inducible expression vectors. This have made L. lactis a desirable and promising host on par with other well established model bacterial or yeast systems such as Escherichia coli, Saccharomyces [corrected] cerevisiae and Bacillus subtilis. In this article, we review recent technological advancements, challenges, future prospects and current diversified examples on the use of L. lactis as a microbial cell factory. Additionally, we will also highlight latest medical-based applications involving whole-cell L. lactis as a live delivery vector for the administration of therapeutics against both communicable and non-communicable diseases.

Matched MeSH terms: Probiotics/therapeutic use
Fulltext A review on the effects of probiotics and antibiotics towards Clostridium difficile infections

Hazirah, A., Loong, Y.Y., Rushdan, A.A., Rukman, A.H., Yazid, M.M.

Pertanika Journal of Science & Technology, 2013;21(2):293-302.
MyJurnal

Clostridium difficile can cause severe diseases with significant morbidity and mortality in infected patients.
The rate of Clostridium difficile infection is high in North America and European countries. Metronidazole
and vancomycin have been recommended as the treatments of choice since 1990s. Recurrent infection
due to Clostridium difficile is common after several days of antibiotic administration. Probiotics have
been used in these patients as an adjunct treatment with some successful findings. However, a detailed
investigation on the use of probiotic for infected patients is still needed, particularly for its real efficacy.

Matched MeSH terms: Probiotics
A standard quantitative method to measure acid tolerance of probiotic cells

Chan ES, Lee PP, Ravindra P, Krishnaiah K, Voo WP

Appl Microbiol Biotechnol, 2010 Mar;86(1):385-91.
PMID: 20033402 DOI: 10.1007/s00253-009-2384-y

The aim of this work was to develop a standard quantitative method to measure the acid tolerance of probiotic cells when exposed to a simulated gastric fluid. Three model strains of different cell concentrations were exposed to a standard simulated gastric fluid of fixed volume. The fluid pH ranged from pH 1.5 to 2.5. In general, the death kinetics followed an exponential trend. The overall death constant, k (d), for all strains was found to be in a power relationship with the pH value and the initial cell concentration, and it can be expressed as k(d)=k(AII) (pH(-9.0)N(0)(-0.19)) where k (AII) is defined as the acid intolerance indicator and N (0) is the initial cell concentration (CFU/ml). This equation was validated with the experimental data with an average R (2) of 0.98. The acid intolerance of cells can be quantitatively expressed by the k (AII) values, where higher value indicates higher intolerance. In conclusion, a standard quantitative method has been developed to measure the acid tolerance of probiotic cells. This could facilitate the selection of probiotic strains and processing technologies.

Matched MeSH terms: Probiotics*
Adhesion ability and cytotoxic evaluation of lactobacillus strains isolated from malaysian fermented fish (pekasam) on ht-29 and ccd-18co intestinal cells

Ida Muryany, Ahmad Rohi Ghazali, Nor Fadilah Rajab, Hing HL, Ina-Salwany, Mohd Zamri Saad, et al.

Sains Malaysiana, 2018;47:2391-2399.

Bacterial adhesion to host cells is the most important probiotic character. However, the adhesion of probiotic should not
affect the viability of the host cells. In this study, Lactobacillus plantarum strain L8, Lactobacillus plantarum strain L20
and Lactobacillus pentosus strain S1 were tested for their cytotoxic effects through MTT assay and their ability to adhere
and colonize on HT-29 and CCD-18Co intestinal cells as detected microscopically using light microscopy and Scanning
Electron Microscopy (SEM). No cytotoxicity effects were observed on both intestinal cells following 24 h treatment with
all Lactobacillus strains. Additionally, all strains demonstrated strong adhesive activity where more than 100 bacteria
adhered to both intestinal cells although differences in the adhesion scores observed among different strains. The adhesion
as observed via SEM showed an autoagreggative pattern and adhered as clusters on the surface of both intestinal cells.
In conclusion, all three Lactobacillus strains are non-cytotoxic to both cells with strong adhesion ability on intestinal
cells and this study also proved that Malaysian fermented fish are good source of probiotic bacteria.

Matched MeSH terms: Probiotics
Administration of Bacillus subtilis strains in the rearing water enhances the water quality, growth performance, immune response, and resistance against Vibrio harveyi infection in juvenile white shrimp, Litopenaeus vannamei

Zokaeifar H, Babaei N, Saad CR, Kamarudin MS, Sijam K, Balcazar JL

Fish Shellfish Immunol, 2014 Jan;36(1):68-74.
PMID: 24161773 DOI: 10.1016/j.fsi.2013.10.007

In this study, vegetative cell suspensions of two Bacillus subtilis strains, L10 and G1 in equal proportions, was administered at two different doses 10(5) (BM5) and 10(8) (BM8) CFU ml(-1) in the rearing water of shrimp (Litopenaeus vannamei) for eight weeks. Both probiotic groups showed a significant reduction of ammonia, nitrite and nitrate ions under in vitro and in vivo conditions. In comparison to untreated control group, final weight, weight gain, specific growth rate (SGR), food conversion ratio (FCR) and digestive enzymatic activity were significantly greater in the BM5 and BM8 groups. Significant differences for survival were recorded in the BM8 group as compared to the control. Eight weeks after the start of experiment, shrimp were challenged with Vibrio harveyi. Statistical analysis revealed significant differences in shrimp survival between probiotic and control groups. Cumulative mortality of the control group was 80%, whereas cumulative mortality of the shrimp that had been given probiotics was 36.7% with MB8 and 50% with MB5. Subsequently, real-time RT-PCR was employed to determine the mRNA levels of prophenoloxidase (proPO), peroxinectin (PE), lipopolysaccharide- and β-1,3-glucan- binding protein (LGBP) and serine protein (SP). The expression of all immune-related genes studied was only significantly up-regulated in the BM5 group compared to the BM8 and control groups. These results suggest that administration of B. subtilis strains in the rearing water confers beneficial effects for shrimp aquaculture, considering water quality, growth performance, digestive enzymatic activity, immune response and disease resistance.

Matched MeSH terms: Probiotics/pharmacology*
Ageing and the gut-brain axis: lessons from the Drosophila model

Tan FHP, Shamsuddin S, Zainuddin A

Benef Microbes, 2023 Dec 13;14(6):591-607.
PMID: 38350490 DOI: 10.1163/18762891-20230056

The steady decline of physiological function and increased vulnerability to age-related disorders are two features of the complicated biological process of ageing. As a key organ for nutrient absorption, metabolism, and immunological regulation, the gut plays a major part in the ageing process. Drosophila melanogaster, a well-established model organism, has emerged as a significant tool for exploring the intricate rapport between the gut and ageing. Through the use of Drosophila models, the physiological and molecular elements of the gut-brain axis have been thoroughly explored. These models have also provided insights into the mechanisms by which gut health impacts ageing and age-related illnesses. Drosophila's gut microbiota experience dysbiosis with age which has been linked to age-related diseases. To prevent this and promote healthy ageing in Drosophila, gut microbiota modification methods, such as dietary restriction in tandem with time-restricted feeding, administration of pro-, pre- and synbiotics, as well as pharmaceutical interventions have been generated with positive impacts. The article also covers the drawbacks and difficulties of investigating the gut via the Drosophila. Thus, with an emphasis on the lessons discovered from Drosophila research, this review provides an extensive description of the current studies on the role of the gut-brain axis in ageing and health.

Matched MeSH terms: Probiotics*
Agrowaste-based nanofibers as a probiotic encapsulant: fabrication and characterization

Fung WY, Yuen KH, Liong MT

J Agric Food Chem, 2011 Aug 10;59(15):8140-7.
PMID: 21711050 DOI: 10.1021/jf2009342

This study explored the potential of soluble dietary fiber (SDF) from agrowastes, okara (soybean solid waste), oil palm trunk (OPT), and oil palm frond (OPF) obtained via alkali treatment, in the nanoencapsulation of Lactobacillus acidophilus . SDF solutions were amended with 8% poly(vinyl alcohol) to produce nanofibers using electrospinning technology. The spinning solution made from okara had a higher pH value at 5.39 ± 0.01 and a higher viscosity at 578.00 ± 11.02 mPa·s (P < 0.05), which resulted in finer fibers. FTIR spectra of nanofibers showed the presence of hemicellulose material in the SDF. Thermal behavior of nanofibers suggested possible thermal protection of probiotics in heat-processed foods. L. acidophilus was incorporated into the spinning solution to produce nanofiber-encapsulated probiotic, measuring 229-703 nm, visible under fluorescence microscopy. Viability studies showed good bacterial survivability of 78.6-90% under electrospinning conditions and retained viability at refrigeration temperature during the 21 day storage study.

Matched MeSH terms: Probiotics/chemistry*
Allantoin, a Potential Metabolite That Promotes AMPK Phosphorylation and Suppresses Cholesterol Biosynthesis Via the Mevalonate Pathway and Bloch Pathway

Yap PG, Choi SB, Liong MT

Appl Biochem Biotechnol, 2020 May;191(1):226-244.
PMID: 32125649 DOI: 10.1007/s12010-020-03265-2

This study aimed to evaluate the effect of probiotic administration on obese and ageing models. Sprague Dawley rats were subjected to high-fat diet (HFD) and injected with D-galactose to induce premature ageing. Upon 12 weeks of treatment, the faecal samples were collected and subjected to gas chromatography-mass spectrophotometry (GC-MS) analysis for metabolite detection. The sparse partial least squares discriminant analysis (sPLS-DA) showed a distinct clustering pattern of metabolite profile in the aged and obese rats administered with probiotics Lactobacillus plantarum DR7 and L. reuteri 8513d, particularly with a significantly higher concentration of allantoin. Molecular docking simulation showed that allantoin promoted the phosphorylation (activation) of adenosine monophosphate-activated kinase (AMPK) by lowering the substrate free energy of binding (FEB) and induced the formation of an additional hydrogen bond between Val184 and the substrate AMP. Allantoin also suppressed cholesterol biosynthesis by either inducing enzyme inhibition, occupying or blocking the putative binding site to result in non-spontaneous substrate binding, as in the cases of 3-hydroxy-methylglutaryl-coA reductase (HMGCR), mevalonate kinase (MVK) and lanosterol demethylase (LDM) where positive FEBs were reported. These results demonstrated the potential of allantoin to alleviate age-related hypercholesterolaemia by upregulating AMPK and downregulating cholesterol biosynthesis via the mevalonate pathway and Bloch pathway.

Matched MeSH terms: Probiotics/pharmacology*
Fulltext Alteration of the Gut Microbiome in Normal and Overweight School Children from Selangor with Lactobacillus Fermented Milk Administration

Joseph N, Clayton JB, Hoops SL, Linhardt CA, Mohd Hashim A, Mohd Yusof BN, et al.

Evol Bioinform Online, 2020;16:1176934320965943.
PMID: 33281440 DOI: 10.1177/1176934320965943

Childhood obesity is a serious public health problem worldwide. Perturbations in the gut microbiota composition have been associated with the development of obesity in both children and adults. Probiotics, on the other hand, are proven to restore the composition of the gut microbiome which helps reduce the development of obesity. However, data on the effect of probiotics on gut microbiota and its association with childhood obesity is limited. This study aims to determine the effect of probiotics supplement intervention on gut microbiota profiles in obese and normal-weight children. A total of 37 children, 17 normal weight, and 20 overweight school children from a government school in Selangor were selected to participate in this study. Participants were further divided into intervention and control groups. The intervention groups received daily probiotic drinks while the control groups continued eating their typical diet. Fecal samples were collected from the participants for DNA extraction. The hypervariable V3 and V4 regions of 16S rRNA gene were amplified and sequenced using the Illumina MiSeq platform. No significant differences in alpha diversity were observed between normal weight and obese children in terms of the Shannon Index for evenness or species richness. However, a higher intervention effect on alpha diversity was observed among normal-weight participants compared to obese. The participants' microbiome was found to fluctuate throughout the study. Analysis of the taxa at species level showed an increase in Bacteroides ovatus among the normal weight cohort. Genus-level comparison revealed a rise in genus Lachnospira and Ruminococcus in the overweight participants after intervention, compared to the normal-weight participants. The probiotics intervention causes an alteration in gut microbiota composition in both normal and overweight children. Though the association could not be defined statistically, this study has provided an improved understanding of the intervention effect of probiotics on gut microbiome dysbiosis in an underrepresented population.

Matched MeSH terms: Probiotics
Anaerobic Probiotics: The Key Microbes for Human Health

El Enshasy H, Malik K, Malek RA, Othman NZ, Elsayed EA, Wadaan M

Adv. Biochem. Eng. Biotechnol., 2016;156:397-431.
PMID: 26907552

Human gastrointestinal microbiota (HGIM) incorporate a large number of microbes from different species. Anaerobic bacteria are the dominant organisms in this microbial consortium and play a crucial role in human health. In addition to their functional role as the main source of many essential metabolites for human health, they are considered as biotherapeutic agents in the regulation of different human metabolites. They are also important in the prevention and in the treatment of different physical and mental diseases. Bifidobacteria are the dominant anaerobic bacteria in HGIM and are widely used in the development of probiotic products for infants, children and adults. To develop bifidobacteria-based bioproducts, therefore, it is necessary to develop a large-scale biomass production platform based on a good understanding of the ideal medium and bioprocessing parameters for their growth and viability. In addition, high cell viability should be maintained during downstream processing and storage of probiotic cell powder or the final formulated product. In this work we review the latest information about the biology, therapeutic activities, cultivation and industrial production of bifidobacteria.

Matched MeSH terms: Probiotics/metabolism*; Probiotics/therapeutic use*
Angiotensin I-converting enzyme inhibitory activity and bioconversion of isoflavones by probiotics in soymilk supplemented with prebiotics

Yeo SK, Liong MT

Int J Food Sci Nutr, 2010 Mar;61(2):161-81.
PMID: 20085504 DOI: 10.3109/09637480903348122

Lactobacillus sp. FTDC 2113, L. acidophilus FTDC 8033, L. acidophilus ATCC 4356, L. casei ATCC 393, Bifidobacterium FTDC 8943 and B. longum FTDC 8643 were incorporated into soymilk supplemented with fructooligosaccharides (FOS), inulin, mannitol, maltodextrin and pectin. The objective of the present study was to evaluate the effects of prebiotics on the bioactivity of probiotic-fermented soymilk. Proteolytic activity was increased in the presence of FOS, while the supplementation of inulin and pectin increased the angiotensin I-converting enzyme inhibitory activity accompanied by lower IC(50) values. The beta-glucosidase activity was also enhanced in the presence of pectin. This led to higher bioconversion of glucosides to aglycones by probiotics, especially genistin and malonyl genistin to genistein. Results from this study indicated that the supplementation of prebiotics enhanced the in-vitro antihypertensive effect and production of bioactive aglycones in probiotic-fermented soymilk. Therefore, this soymilk could potentially be used as a dietary therapy to reduce the risks of hypertension and hormone-dependent diseases such as breast cancer, prostate cancer and osteoporosis.

Matched MeSH terms: Probiotics*
Anti-breast cancer effects of live, heat-killed and cytoplasmic fractions of Enterococcus faecalis and Staphylococcus hominis isolated from human breast milk

Hassan Z, Mustafa S, Rahim RA, Isa NM

In Vitro Cell Dev Biol Anim, 2016 Mar;52(3):337-348.
PMID: 26659392 DOI: 10.1007/s11626-015-9978-8

Development of tumour that is resistant to chemotherapeutics and synthetic drugs, coupled with their life-threatening side effects and the adverse effects of surgery and hormone therapies, led to increased research on probiotics' anticancer potentials. The current study investigated the potential of live, heat-killed cells (HKC) and the cytoplasmic fractions (CF) of Enterococcus faecalis and Staphylococcus hominis as anti-breast cancer agents. MCF-7 cell line was treated with 25, 50, 100 and 200 μg/mL each of live, HKC and CF of the bacteria; and cytotoxicity was evaluated for 24, 48 and 72 h using MTT assay. The morphological features of the treated cells were examined by fluorescence microscopy. The stage of cell cycle arrest and apoptosis were quantified by flow cytometry. The bacterial effect on non-malignant breast epithelial cell line, MCF-10A, was assessed using MTT assay for 24, 48 and 72 h. All the three forms of the bacteria caused a significant decrease in MCF-7 (up to 33.29%) cell proliferation in concentration- and time-dependent manner. Morphological features of apoptosis like cell death, cell shrinkage and membrane blebbing were observed. Flow cytometry analyses suggested that about 34.60% of treated MCF-7 was undergoing apoptosis. A strong anti-proliferative activity was efficiently induced through sub-G1 accumulation (up to 83.17%) in treated MCF-7 and decreased number in the G0/G1 phase (74.39%). MCF-10A cells treated with both bacteria showed no significant difference with the untreated (>90% viability). These bacteria can be used as good alternative nutraceutical with promising therapeutic indexes for breast cancer because of their non-cytotoxic effects to normal cells.

Matched MeSH terms: Probiotics
Antimicrobial and bactericidal impacts of Bacillus amyloliquefaciens CECT 5940 on fecal shedding of pathogenic bacteria in dairy calves and adult dogs

Vazquez-Mendoza P, Elghandour MMM, Alaba PA, Sánchez-Aparicio P, Alonso-Fresán MU, Barbabosa-Pliego A, et al.

Microb Pathog, 2018 Jan;114:458-463.
PMID: 29180295 DOI: 10.1016/j.micpath.2017.11.040

Two experiments were carried out to evaluate the bactericidal impacts of Bacillus amyloliquefaciens CECT 5940 on the shedding of faecal pathogenic bacteria in dairy calves (Experiment 1) and in adults dogs (experiment 2). In the calves experiment, a completely randomized design was used to investigate the faecal bacteria profile of Holstein dairy calves fed with either pasteurized waste milk (PWM; n = 9) or a formulated non-medicated milk replacer (NMR; n = 9) for 60 d. The NMR containing sodium-butyrate and the active probiotic B. amyloliquefaciens CECT 5940. In the dogs experiment, addition of same probiotic (i.e., B. amyloliquefaciens CECT 5940) was carried out in two stages. The first stage started from day 7-37, and the second from day 44-71. The assessment of faecal score measured on day 22, 37, 42, 57, 71 and 77 to determine the texture of the stools. Calves received PWM consumed (P

Matched MeSH terms: Probiotics*
Antimicrobial and immunoregulatory effects of Lactobacillus delbrueckii 45E against genitourinary pathogens

Bnfaga AA, Lee KW, Than LTL, Amin-Nordin S

J Biomed Sci, 2023 Mar 23;30(1):19.
PMID: 36959635 DOI: 10.1186/s12929-023-00913-7

BACKGROUND: Lactobacilli are essential microbiota that maintain a healthy, balanced vaginal environment. Vaginitis is a common infection in women during their reproductive years. Many factors are associated with vaginitis; one of them is the imbalance of microbiota in the vaginal environment. This study aimed to evaluate the antimicrobial properties of Lactobacillus delbrueckii 45E (Ld45E) against several species of bacteria, namely, Group B Streptococcus (GBS), Escherichia coli, Klebsiella spp., and Candida parapsilosis, as well as to determine the concentration of interleukin-17 (IL-17) in the presence of Ld45E.
METHODS: The probiotic characteristics of Ld45E were evaluated by examining its morphology, pH tolerance, adhesive ability onto HeLa cells, hemolytic activity, antibiotic susceptibility, and autoaggregation ability. Then, the antimicrobial activity of Ld45E was determined using Ld45E culture, cell-free supernatant, and crude bacteriocin solution. Co-aggregation and competition ability assays against various pathogens were conducted. The immunoregulatory effects of Ld45E were analyzed by measuring the proinflammatory cytokine IL-17. A p-value less than 0.05 was considered statistical significance.
RESULTS: Ld45E is 3-5 mm in diameter and round with a flat-shaped colony. pH 4 and 4.5 were the most favorable range for Ld45E growth within 12 h of incubation. Ld45E showed a strong adhesion ability onto HeLa cells (86%) and negative hemolytic activities. Ld45E was also sensitive to ceftriaxone, cefuroxime, ciprofloxacin, and doxycycline. We found that it had a good autoaggregation ability of 80%. Regarding antagonistic properties, Ld45E culture showed strong antimicrobial activity against GBS, E. coli, and Klebsiella spp. but only a moderate effect on C. parapsilosis. Cell-free supernatant of Ld45E exerted the most potent inhibitory effects at 40 °C against all genital pathogens, whereas bacteriocin showed a robust inhibition at 37 °C and 40 °C. The highest co-aggregation affinity was observed with GBS (81%) and E. coli (40%). Competition ability against the adhesion of GBS (80%), E. coli (76%), Klebsiella (72%), and C. parapsilosis (58%) was found. Ld45E was able to reduce the induction of the proinflammatory protein IL-17.
CONCLUSIONS: Ld45E possessed antimicrobial and immunoregulatory properties, with better cell-on-cell activity than supernatant activity. Thus, Ld45E is a potential probiotic candidate for adjunct therapy to address vaginal infections.

Matched MeSH terms: Probiotics*
Association of body mass index and intestinal (faecal) Streptococcus in adults in Xining city, China P.R

Ma M, Su J, Wang Y, Wang L, Li Y, Ding G, et al.

Benef Microbes, 2022 Dec 07;13(6):465-472.
PMID: 36264094 DOI: 10.3920/BM2021.0046

Body mass index (BMI) and gut microbiota show significant interaction, but most studies on the relationship between BMI and gut microbiota have been done in Western countries. Relationships that are also identified in other cultural backgrounds are likely to have functional importance. Hence here we explore gut microbiota in adults living in Xining city (China P.R.) and relate results to subject BMI. Analysis of bacterial 16s rRNA gene was performed on faecal samples from participants with normal-weight (n=24), overweight (n=24), obesity (n=11) and type 2 diabetes (T2D) (n=8). The results show that unweighted but not weighted Unifrac distance was significantly different when gut microbiota composition was compared between the groups. Importantly, the genus Streptococcus was remarkably decreased in both obese subjects and subjects suffering from T2D, as compared to normal-weight subjects. Accordingly, strong association was identified between the genus Streptococcus and BMI and especially Streptococcus salivarius subsp. thermophiles was a major contributor in this respect. As previous studies have shown that Streptococcus salivarius subsp. thermophiles is also negatively associated with obesity in Western cohorts, our results suggest that this species is a potential probiotic for the prevention of obesity and related disorders.

Matched MeSH terms: Probiotics*

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