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Viability, Acid and Bile Tolerance of Spray Dried Probiotic Bacteria and Some Commercial Probiotic Supplement Products Kept at Room Temperature

Dianawati D, Mishra V, Shah NP

J Food Sci, 2016 Jun;81(6):M1472-9.
PMID: 27145163 DOI: 10.1111/1750-3841.13313

Production of probiotic food supplements that are shelf-stable at room temperature has been developed for consumer's convenience, but information on the stability in acid and bile environment is still scarce. Viability and acid and bile tolerance of microencapsulated Bifidobacterium spp. and Lactobacillus acidophilus and 4 commercial probiotic supplements were evaluated. Bifidobacterium and L. acidophilus were encapsulated with casein-based emulsion using spray drying. Water activity (aw ) of the microspheres containing Bifidobacterium or L. acidophilus (SD GM product) was adjusted to 0.07 followed by storage at 25 °C for 10 wk. Encapsulated Bifidobacterium spp. and Lactobacillus acidophilus and 4 commercial probiotic supplement products (AL, GH, RE, and BM) were tested. Since commercial probiotic products contained mixed bacteria, selective media MRS-LP (containing L-cysteine and Na-propionate) and MRS-clindamycin agar were used to grow Bifidobacterium spp. or L. acidophilus, respectively, and to inhibit the growth of other strains. The results showed that aw had a strong negative correlation with the viability of dehydrated probiotics of the 6 products. Viable counts of Bifidobacterium spp. and L. acidophilus of SD GM, AL, and GH were between 8.3 and 9.2 log CFU/g, whereas that of BM and RE were between 6.7 and 7.3 log CFU/g. Bifidobacterium in SD GM, in AL, and in GH products and L. acidophilus in SD GM, in AL, and in BM products demonstrated high tolerance to acid. Most of dehydrated probiotic bacteria were able to survive in bile environment except L. acidophilus in RE product. Exposure to gastric juice influenced bacterial survivability in subsequent bile environment.

Matched MeSH terms: Bile Acids and Salts/pharmacology
Fulltext Bile acids and their respective conjugates elicit different responses in neonatal cardiomyocytes: role of Gi protein, muscarinic receptors and TGR5

Ibrahim E, Diakonov I, Arunthavarajah D, Swift T, Goodwin M, McIlvride S, et al.

Sci Rep, 2018 05 08;8(1):7110.
PMID: 29740092 DOI: 10.1038/s41598-018-25569-4

Bile acids are recognised as bioactive signalling molecules. While they are known to influence arrhythmia susceptibility in cholestasis, there is limited knowledge about the underlying mechanisms. To delineate mechanisms underlying fetal heart rhythm disturbances in cholestatic pregnancy, we used FRET microscopy to monitor cAMP release and contraction measurements in isolated rodent neonatal cardiomyocytes. The unconjugated bile acids CDCA, DCA and UDCA and, to a lesser extent, CA were found to be relatively potent agonists for the GPBAR1 (TGR5) receptor and elicit cAMP release, whereas all glyco- and tauro- conjugated bile acids are weak agonists. The bile acid-induced cAMP production does not lead to an increase in contraction rate, and seems to be mediated by the RI isoform of adenylate cyclase, unlike adrenaline-dependent release which is mediated by the RII isoform. In contrast, bile acids elicited slowing of neonatal cardiomyocyte contraction indicating that other signalling pathways are involved. The conjugated bile acids were found to be partial agonists of the muscarinic M2, but not sphingosin-1-phosphate-2, receptors, and act partially through the Gi pathway. Furthermore, the contraction slowing effect of unconjugated bile acids may also relate to cytotoxicity at higher concentrations.

Matched MeSH terms: Bile Acids and Salts/metabolism*
Cholesterol homeostasis associated with probiotic supplementation in vivo

Lim PS, Loke CF, Ho YW, Tan HY

J Appl Microbiol, 2020 Nov;129(5):1374-1388.
PMID: 32356362 DOI: 10.1111/jam.14678

AIMS: To determine the mechanism underlying the serum cholesterol reduction effect by probiotics isolated from local fermented tapioca (Tapai).
METHODS AND RESULTS: Lactic acid bacteria strains were isolated and examined for acid tolerance, bile salt resistance and hypocholesterolemic properties. Among the isolates, Lactobacillus plantarum TAR4 showed the highest cholesterol reduction ability (48·01%). The focus in the in vivo trial was to elucidate the cholesterol balance from findings pertaining to serum cholesterol reduction in rat model fed with high fat diet via oral administration. Rats fed with high-cholesterol diet supplemented with Lact. plantarum TAR4 showed significant reduction in serum total cholesterol (29·55%), serum triglyceride (45·31%) and liver triglyceride (23·44%) as compared to high-cholesterol diet (HCD) group. There was a significant increment in faecal triglyceride (45·83%) and faecal total bile acid (384·95%) as compared to HCD group.
CONCLUSIONS: The findings showed that probiotic Lact. plantarum TAR4 supplementation reduced the absorption of bile acids for enterohepatic recycling and increased the catabolism of cholesterol to bile acids and not by suppressing the rate of cholesterol synthesis.
SIGNIFICANCE AND IMPACT OF STUDY: Probiotic supplements could provide a new nonpharmacological alternative to reduce cardiovascular risk factors.

Matched MeSH terms: Bile Acids and Salts/metabolism
Hypocholesterolemic Effects of Kenaf Seed Oil, Macroemulsion, and Nanoemulsion in High-Cholesterol Diet Induced Rats

Cheong AM, Jessica Koh JX, Patrick NO, Tan CP, Nyam KL

J Food Sci, 2018 Mar;83(3):854-863.
PMID: 29412455 DOI: 10.1111/1750-3841.14038

This study aimed to evaluate the effect of kenaf seed oil (KSO), kenaf seed oil-in-water macroemulsion (KSOM), kenaf seed oil-in-water nanoemulsions (KSON), and emulsifier mixtures (EM) on serum lipid profile, liver oxidative status, and histopathological changes in high-cholesterol fed rats. Stability and characteristic of KSOM and KSON were carried out prior to in vivo study. Forty-two Sprague-Dawley rats were divided into 7 groups (6 rats each) and induced hypercholesterolemia by feeding high cholesterol diet (HCD) for 14 days prior to treatments. Different treatments were introduced on day 15 to 29 while supplemented with HCD and removal of HCD during treatment on day 30 to 43, except for HCD group. Body weight and serum lipid profiles were measured at 3 different points: after hypercholesterolemia was induced, on day 29, and at the end of the experiment. Relative liver weight, atherogenic index, coronary risk index, and fecal total bile acids were also determined at the end of experiment. KSON showed significantly higher stability than KSOM and FTIR exhibited good encapsulation of KSO after 1.5 years of storage. Serum total cholesterol, low density lipoprotein cholesterol, lipid peroxidation levels in HCD group without treatment were significantly higher compared to normal control group and all treatment groups. All samples demonstrated hypocholesterolemic effect, but KSON exhibited higher efficiency in cholesterol-lowering properties, weight control and decreased liver fat as confirmed by histopathological evaluation. The overall results revealed that the efficacy of different treatments was in descending order of KSON, KSO, KSOM, and EM.
PRACTICAL APPLICATION: Kenaf seed oil-in-water nanoemulsion (KSON) has the potential to be used as a natural alternative to the synthetic hypocholesterolemic drug in the future. However, larger sample size and clinical trial are needed to confirm on this potential application. In addition, treatment with KSON was suggested to prevent cardiovascular disease and fatty liver.

Matched MeSH terms: Bile Acids and Salts/metabolism
Fulltext Familial hypercholesterolaemia: an updated overall management

Noor Alicezah Mohd Kasim, Chua Yung An, Hapizah Nawawi

Journal of Clinical and Health Sciences, 2020;5(2):19-38.
MyJurnal

Familial hypercholesterolaemia (FH), the commonest and serious but potentially treatable
form of inherited dyslipidaemias, is characterised by severely elevated plasma low-density
lipoprotein-cholesterol (LDL-C) level, which subsequently leads to premature coronary artery
disease (pCAD). Effectiveness of FH early detection and treatment is supported by the
outcome of several international cohort studies. Optimal FH management relies on
prescription of statins either alone or together with other lipid-lowering therapies (LLT).
Intensive lifestyle intervention is required in parallel with LLT, which should be commenced at
diagnosis in adults and childhood. Treatment with high intensity statin should be started as
soon as possible. Combination with ezetimibe and/or bile acid sequestrants is indicated if
target LDL-C is not achieved. For FH patients in the very-high risk category, if their LDL-C
targets are not achieved, despite being on maximally tolerated statin dose and ezetimibe,
proprotein convertase subtilisin/kexin type1 inhibitor (PCSK9i) is recommended. In statin
intolerance, ezetimibe alone, or in combination with PCSK9i may be considered. Clinical
evaluation of response to treatment and safety are recommended to be done about 4-6 weeks
following initiation of treatment. Homozygous FH (HoFH) patients should be treated with
maximally tolerated intensive LLT and, when available, with lipoprotein apheresis. This review
highlights the overall management, and optimal treatment combinations in FH in adults and
children, newer LLT including PCSK9i, microsomal transfer protein inhibitor, allele-specific
oligonucleotide to ApoB100 and PCSK9 mRNA. Family cascade screening and/or screening
of high-risk individuals, is the most cost-effective way of identifying FH cases and initiating
early and adequate LLT.

Matched MeSH terms: Bile Acids and Salts
Fulltext Screening of pathogenic bacteria from sea cucumber Acaudina molpadioides in Pulau Langkawi, Kedah

Abigail Li Yen Lew, Nurzafirah Mazlan, Siti Marwanis Anua, Thung Tze Young

Malaysian Journal of Medicine and Health Sciences, 2020;16(3):42-46.
MyJurnal

Introduction: The outbreaks of foodborne diseases have been linked to the consumption of contaminated seafood. This research aims to screen the bacteria from the sea cucumbers Acaudina molpadioides collected from Pulau Langkawi. Methods: A total of 22 sea cucumber samples were collected randomly from Pulau Langkawi, Kedah, Malaysia. The samples were isolated and identified for the presence of bacteria using the conventional culture-based method. Presumptive bacteria colonies were subjected to various biochemical and antimicrobial susceptibility tests. Results: There were no bacterial growth in Hektoen Enteric (HE) agar and Thiosulphate-Citrate-Bile Salt (TCBS) agar. Positive samples were isolated from MacConkey (MAC) agar with 6 samples were Staphylococcus spp. (27.27%), 14 samples were Proteus spp. (63.63%) and 2 samples were Bacillus spp. (9.01%). Among these isolates, highest resistance was found against Ampicillin (45%) followed by Tetracycline (40%). Conclusion: The results indicate that the sea cucumbers Acaudina molpadioides were contaminated with potential bacteria. There is a need for adequate consumer protection measures.

Matched MeSH terms: Bile Acids and Salts
Fulltext CHARACTERISATION OF LACTIC ACID BACTERIA ISOLATED FROM KEFIR MILK MADE FROM DAIRY AND NON-DAIRY SOURCES AND THEIR SENSORY ACCEPTANCE

CHAN KHER ZHI, MOHD NIZAM LANI, MOHD NIZAM LANI, YUSNITA HAMZAH, FAUZIAH TUFAIL AHMAD, HAFIZAH NIK UBAIDILLAH

UMT Journal of Undergraduate Research, 2021;3(2):37-50.
MyJurnal

Kefir is a fermented milk obtained by fermenting milk with kefir grains. The chemical composition ofdairy and non-dairy sources may affect the growth and characterisation of lactic acid bacteria (LAB). In this study, different sources of milk (cow milk) and non-dairy milk (soymilk and coconut milk) were used as the fermentation media for kefir products. The objectives of the study were to isolate and characterise LAB from kefir drink produced from dairy and non-dairy milk. LAB was isolated using different cultural methods, such as MRS Agar, MRS with 0.8% CaCO3, and M17 Agar. The characteristics of the LAB isolates were determined using morphological, biochemical tests and the API 50 CHL kit. The physicochemical composition of the samples was determined using titratable acidity and pH level. Sensory evaluation of the kefir drink samples was also carried out. Results confirmed that the isolates were identified as Lactobacillus buchneri, Lactobacillus brevis 1, Leuconostoc mesenteroides, Lactobacillus acidophilus 3and Lactobacillus plantarum 1. The L. buchneri, L. brevis, Leu. mesenteroides and L. acidophilus are heterofermentative bacteria, whereas L. plantarum is a homofermentative bacterium. Four LAB isolates have the potential to be used as probiotic strains due to their high resistant to low pH and bile salt. The sensory scores of these products range between 5.00 and 8.00 in the 9-point hedonic scale. Most of the sensory panelists preferred cow milk kefir (p < 0.05) compared with coconut milk kefir and soy milk kefir during the sensory evaluation of all attributes. Meanwhile, the preference between coconut milk kefir and soy milk kefir was similar (p>0.05) in all attributes. Therefore, this study will be useful for probiotic manufacturers in the production of alternative probiotic drinks using dairy and non-dairy milk.

Matched MeSH terms: Bile Acids and Salts
Fulltext Heart and bile acids - Clinical consequences of altered bile acid metabolism

Vasavan T, Ferraro E, Ibrahim E, Dixon P, Gorelik J, Williamson C

Biochim Biophys Acta Mol Basis Dis, 2018 04;1864(4 Pt B):1345-1355.
PMID: 29317337 DOI: 10.1016/j.bbadis.2017.12.039

Cardiac dysfunction has an increased prevalence in diseases complicated by liver cirrhosis such as primary biliary cholangitis and primary sclerosing cholangitis. This observation has led to research into the association between abnormalities in bile acid metabolism and cardiac pathology. Approximately 50% of liver cirrhosis cases develop cirrhotic cardiomyopathy. Bile acids are directly implicated in this, causing QT interval prolongation, cardiac hypertrophy, cardiomyocyte apoptosis and abnormal haemodynamics of the heart. Elevated maternal serum bile acids in intrahepatic cholestasis of pregnancy, a disorder which causes an impaired feto-maternal bile acid gradient, have been associated with fatal fetal arrhythmias. The hydrophobicity of individual bile acids in the serum bile acid pool is of relevance, with relatively lipophilic bile acids having a more harmful effect on the heart. Ursodeoxycholic acid can reverse or protect against these detrimental cardiac effects of elevated bile acids.

Matched MeSH terms: Bile Acids and Salts/blood; Bile Acids and Salts/metabolism*; Bile Acids and Salts/chemistry
Fulltext Bile Sensing: The Activation of Vibrio parahaemolyticus Virulence

Letchumanan V, Chan KG, Khan TM, Bukhari SI, Ab Mutalib NS, Goh BH, et al.

Front Microbiol, 2017;8:728.
PMID: 28484445 DOI: 10.3389/fmicb.2017.00728

Bacteria must develop resistance to various inhospitable conditions in order to survive in the human gastrointestinal tract. Bile, which is secreted by the liver, and plays an important role in food digestion also has antimicrobial properties and is able to disrupt cellular homeostasis. Paradoxically, although bile is one of the guts defenses, many studies have reported that bacteria such as Vibrio parahaemolyticus can sense bile and use its presence as an environmental cue to upregulate virulence genes during infection. This article aims to discuss how bile is detected by V. parahaemolyticus and its role in regulating type III secretion system 2 leading to human infection. This bile-bacteria interaction pathway gives us a clearer understanding of the biochemical and structural analysis of the bacterial receptors involved in mediating a response to bile salts which appear to be a significant environmental cue during initiation of an infection.

Matched MeSH terms: Bile Acids and Salts
Different effects of two newly-isolated probiotic Lactobacillus plantarum 15HN and Lactococcus lactis subsp. Lactis 44Lac strains from traditional dairy products on cancer cell lines

Haghshenas B, Abdullah N, Nami Y, Radiah D, Rosli R, Khosroushahi AY

Anaerobe, 2014 Dec;30:51-9.
PMID: 25168457 DOI: 10.1016/j.anaerobe.2014.08.009

Lactobacillus and Lactococcus strains isolated from food products can be introduced as probiotics because of their health-promoting characteristics and non-pathogenic nature. This study aims to perform the isolation, molecular identification, and probiotic characterization of Lactobacillus and Lactococcus strains from traditional Iranian dairy products. Primary probiotic assessments indicated high tolerance to low pH and high bile salt conditions, high anti-pathogenic activities, and susceptibility to high consumption antibiotics, thus proving that both strains possess probiotic potential. Cytotoxicity assessments were used to analyze the effects of the secreted metabolite on different cancer cell lines, including HT29, AGS, MCF-7, and HeLa, as well as a normal human cell line (HUVEC). Results showed acceptable cytotoxic properties for secreted metabolites (40 μg/ml dry weight) of Lactococcus lactis subsp. Lactis 44Lac. Such performance was similar to that of Taxol against all of the treated cancer cell lines; however, the strain exhibited no toxicity on the normal cell line. Cytotoxic assessments through flow cytometry and fluorescent microscopy demonstrated that apoptosis is the main cytotoxic mechanism for secreted metabolites of L. lactis subsp. Lactis 44Lac. By contrast, the effects of protease-treated metabolites on the AGS cell line verified the protein nature of anti-cancer metabolites. However, precise characterizations and in vitro/in vivo investigations on purified proteins should be conducted before these metabolites are introduced as potential anti-cancer therapeutics.

Matched MeSH terms: Bile Acids and Salts/metabolism
Growth, bioconversion of isoflavones and probiotic properties of parent and subsequent passages of Lactobacillus upon ultraviolet radiation

Yeo SK, Liong MT

Int J Food Sci Nutr, 2012 Nov;63(7):821-31.
PMID: 22264088 DOI: 10.3109/09637486.2011.652942

The objective of this study was to evaluate the effects of ultraviolet (UV) radiation (UVB; 90 J/m²) on growth, bioconversion of isoflavones and probiotic properties of parent and subsequent passages of L. casei FTDC 2113. UV radiation significantly enhanced (P < 0.05) the growth of parent cells in mannitol-soymilk fermented at 37°C for 24 h. This had led to an enhanced intracellular and extracellular β-glucosidase activity with a subsequent increase in bioconversion of isoflavones in mannitol-soymilk (P < 0.05). UV radiation also promoted (P < 0.05) the tolerance of parent cells towards acidic condition (pH 2 and 3) and intestinal bile salts (oxgall, taurocholic and cholic acid). In addition, parent treated cells also exhibited better (P < 0.05) adhesion ability to mucin and antimicrobial activity compared to that of the control. All these positive effects of UV radiation were only prevalent in the parent cells without inheritance by first, second and third passage of cells. Although temporary, our results suggested that UV radiation could enhance the bioactive and probiotic potentials of L. casei FTDC 2113, and thus could be applied for the production of probiotic products with enhanced bioactivity.

Matched MeSH terms: Bile Acids and Salts/metabolism
Fulltext Probiotic Properties of Exopolysaccharide-Producing Lactobacillus Strains Isolated from Tempoyak

Khalil ES, Abd Manap MY, Mustafa S, Alhelli AM, Shokryazdan P

Molecules, 2018 Feb 13;23(2).
PMID: 29438288 DOI: 10.3390/molecules23020398

Tempoyak is a functional Malaysian food (an acid-fermented condiment) which is produced from the pulp of the durian (Durio zibethinus) fruit. The current study aimed to isolate and identify potential exopolysaccharide (EPS)-producing Lactobacillus strains from tempoyak for potential use as probiotics. Seven isolates (DUR2, DUR4, DUR5, DUR8, DUR12, DUR18, and DUR20) out of 44 were able to produce EPS, and exhibited resistance to acid and bile salt compared to the reference strains Lactobacillus rhmnosus (ATCC53103) and L. plantarum (ATCC8014). The seven isolated strains belonged to five different species-L. plantarum, L. fermentum, L. crispatus, L. reuteri, and L. pentosus-which were identified using API 50 CHL and 16S rRNA gene sequences (Polymerase chain reaction, PCR - based). The seven strains displayed different ability to produce EPS (100-850 mg/L). Isolates exhibited a high survivability to acid (pH 3.0), bile salts (0.3%), and gastrointestinal tract model (<70%). Results showed that the auto-aggregation and cell surface hydrophobicity ranged from 39.98% to 60.09% and 50.80% to 80.53%, respectively, whereas, the highest co-aggregation value (66.44%) was observed by L. fermentum (DUR8) with Pseudomonas aeruginosa. The isolates showed good inhibitory activity against tested pathogens, high antioxidant activity (32.29% to 73.36%), and good ability to reduce cholesterol (22.55% to 75.15%). Thus, the seven tested strains have value as probiotics.

Matched MeSH terms: Bile Acids and Salts/pharmacology
Fulltext Metabolomics analysis of herb-partitioned moxibustion treatment on rats with diarrhea-predominant irritable bowel syndrome

Lin X, Liu X, Xu J, Cheng KK, Cao J, Liu T, et al.

Chin Med, 2019;14:18.
PMID: 31080495 DOI: 10.1186/s13020-019-0240-2

Background: Irritable bowel syndrome (IBS) is a common functional gastrointestinal disorder, which is commonly treated with antidiarrhoeal, antispasmodics, serotonergic agents or laxative agents. These treatments provide relief for IBS symptoms but may also lead to undesired side effects. Previously, herb-partitioned moxibustion (HPM) treatment has been demonstrated to be effective in ameliorating symptoms of IBS. However, the underlying mechanism of this beneficial treatment is yet to be established. The aim of the current study was to systematically assess the metabolic alterations in response to diarrhea-predominant IBS (IBS-D) and therapeutic effect of HPM.
Methods: Proton nuclear magnetic resonance spectroscopy (1H NMR)-based metabolomics approach was used to investigate fecal and serum metabolome of rat model of IBS-D with and without HPM treatment.
Results: The current results showed that IBS-induced metabolic alterations in fecal and serum sample include higher level of threonine and UDP-glucose together with lower levels of aspartate, ornithine, leucine, isoleucine, proline, 2-hydroxy butyrate, valine, lactate, ethanol, arginine, 2-oxoisovalerate and bile acids. These altered metabolites potentially involve in impaired gut secretory immune system and intestinal inflammation, malabsorption of nutrients, and disordered metabolism of bile acids. Notably, the HPM treatment was found able to normalize the Bristol stool forms scale scores, fecal water content, plasma endotoxin level, and a number of IBS-induced metabolic changes.
Conclusions: These findings may provide useful insight into the molecular basis of IBS and mechanism of the HPM intervention.

Matched MeSH terms: Bile Acids and Salts

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