Isolation and  In Vitro Screening of Lactic Acid Bacteria  from Some Local Brewed Drinks, Rotten Fruits, Pig  Hindgut and their Characterization

The aim of this study was to screen, isolate and characterize Lactic acid bacteria from brewed drinks, certain rotten fruits and hindguts of pigs, using biochemical and Polymerase Chain Reaction (PCR) methods and to compare them with patented probiotics. Briefly, 200 ml each of Local brews kunu zaki, nono and burukutu were purchased separately from five different points and later pooled to form one liter each. Near rotten pineapples (17) and oranges (3) were purchased, and segments of the hindgut (cecum, colon and rectum) of six slaughtered pigs were collected. Bacteria growths were identified according to their morphological and biochemical characteristics. An in vitro fermentation of four grains, namely: sorghum ( Sorghum bicolor ) yellow and red varieties, Millet ( Pennisetum americanum ) and Maize ( Zae mays ) were later performed using the isolated Lactobacillus bacteria from the local brews, fruits and pig hindgut and compared with the patented bacteria. This process was repeated with 0.01ml of an over - night culture of MRS broth concentration containin g one of the two 10 5 and 10 7 Cfu/ml Lab isolate from local drinks, fruits and pig hindgut isolates. Selected fermenters from pig hindgut and burukutu successfully decreased pH of Millet, Maize and Sorghum from 6 to 3.80 and from 6 to 3.92 respectively henc e can be used as fermenters. Molecular confirmation of Lactobacillus species in pig hind gut and burukutu using direct PCR method and the amplification of the genomic DNA showed that the genus level of Lactobacillus was the same for all the isolates and th e homology analysis inferred from the 16S RNA sequence clearly verified that all the strains were Lactobacillus species. L. casei and L. acidiphilus were identified as species type from burukutu . But none from pig hind gut. Therefore it was concluded that Millet, Maize and Sorghum can be effectively used as components of fermented pig diets and that local brewed drink; burukutu and pig hind gut content are good sources of Lactobacillus bacteria f or fermentation of feed for feeding pigs.

Download Full-text

Identification and Characterization Indigenous of Lactobacillus spfrom Bovine Rumen Fluidof Slaughterhouse

KnE Life Sciences ◽

10.18502/kls.v3i6.1182 ◽

2017 ◽

Vol 3 (6) ◽

pp. 549

Author(s):

Tri Nurhajati ◽

Koesnoto Soepranianondo ◽

Widya Paramita Lokapirnasari ◽

Adriana Monica Sahidu

Keyword(s):

Lactic Acid ◽

Nucleotide Sequence ◽

Lactic Acid Bacteria ◽

16S Rdna ◽

Biochemical Characterization ◽

Rumen Fluid ◽

Lactobacillus Rhamnosus ◽

Bovine Rumen ◽

Pcr Method

The discovery and characterisation of indigenous lactic acid bacteria (LAB) are important for diversity microbes as candidate probiotic. This research was aimed to identify lactic acid bacteria isolate from isolation process of local bovine rumen fluid from slaughterhouse in Surabaya Indonesia. Genotypic testing was conducted by analyzing 16S rDNA and biochemical identification. DNA of sample isolate was isolated and then amplified in vitro through the PCR method. Determination of nucleotide sequence of 16S rDNA was performed with sequencing method. The result of nucleotide sequence was than compared with GenBank database. The BLAST was then applied to identify the phylogenetic tree. Based on the biochemical characterization and nucleotide sequences, that isolate was identified as Lactobacillus rhamnosus subsp TG15. The result of this research showed that L.rhamnosus subsp TG15 showed viability bacteria in MRSA as control as much as 1.1 x 108 CFU/ml, mean while in MRSA pH 2, L.rhamnosus subsp TG15 showed its viability as much as 9.3 x 106 CFU/ml. Viability of isolate on bile tolerance 0.3% was 2.4 x 107 CFU/ml). Index antagonist bacteria test on S.aureus showed inhibition diameter as much as 2.0 mm and in antagonist test on E.coli as much as 2.5 mm. Based on the result, it could be concluded that this research found a new strain of lactic acid bacteria, L.rhamnosus subsp TG15 and that isolate has ability as the probiotic candidate. Keywords: L.rhamnosus subsp TG15; survival on acidity; bile salts; S.aureus and E.coli

Download Full-text

Potential of lactic acid bacteria for the reduction of fumonisin exposure in African fermented maize based foods

World Mycotoxin Journal ◽

10.3920/wmj2017.2184 ◽

2017 ◽

Vol 10 (4) ◽

pp. 309-318 ◽

Cited By ~ 2

Author(s):

P. Dawlal ◽

C. Brabet ◽

M.S. Thantsha ◽

E.M. Buys

Keyword(s):

Lactic Acid ◽

Lactic Acid Bacteria ◽

Storage Conditions ◽

Starter Cultures ◽

Lactobacillus Delbrueckii ◽

Lactobacillus Species ◽

Probiotic Properties ◽

Cereal Products ◽

The Stability

Maize, which contributes to a large portion of the African diet and serves as the base substrate for many fermented cereal products, has been reported to be contaminated with fumonisins. This study aimed to evaluate the in vitro ability of predominant lactic acid bacteria (LAB) in African traditional fermented maize based foods (ogi and mahewu) to bind fumonisin B1 (FB1) and B2 (FB2), as well as the stability of the complex at different pH and temperatures, in particular observed during ogi fermentation and under its storage conditions (time, temperature). The percentage of bound fumonisins was calculated after analysing the level of fumonisins not bound to LAB after a certain incubation time, by HPLC. The results revealed the ability of all tested LAB strains to bind both fumonisins, with binding efficiencies varying between strains and higher for FB2. Binding of fumonisins increased with a decrease in pH from 6 to 4 (observed during the ogi fermentation process) and from 4 to 2 (acidic pH in the stomach), and an increase in temperature (from 30 to 37 °C). The percentage of FB1 and FB2 bound to LAB at pH 4 decreased after 6 days of storage at 30 °C for all LAB strains, except for Lactobacillus plantarum (R1096) for which it increased. Lactobacillus species (L. plantarum and Lactobacillus delbrueckii) were the most efficient in binding FB1 and FB2, whereas Pediococcus sp. was less efficient. Therefore, the Lactobacillus strains tested in this study can be recommended as potential starter cultures for African traditional fermented maize based foods having detoxifying and probiotic properties.

Download Full-text

Antimicrobial potential of lactic acid bacteria from domestic chickens (Gallus domesticus) from south Celebes, Indonesia, in different growth phases: in vitro experiments supported by computational docking

Iranian Journal of Microbiology ◽

10.18502/ijm.v12i1.2519 ◽

2020 ◽

Author(s):

Dirayah Rauf Husain ◽

Syahrul Gunawan ◽

Sulfahri Sulfahri

Keyword(s):

Lactic Acid ◽

Lactic Acid Bacteria ◽

Pathogenic Bacteria ◽

Gallus Domesticus ◽

Diffusion Method ◽

Subtilis Strain ◽

Antimicrobial Compounds ◽

E Coli ◽

16S Rna

Background and Objectives: Pathogenic bacterial infection is one of the factors that can cause extensive losses in poultry farming. Pathogenic bacteria that infect domestic chickens (Gallus domesticus) include Escherichia coli. This study has investigated antimicrobial compounds from probiotic bacteria isolated from the digestive tract of domestic chickens origi- nating from Takalar Regency, South Sulawesi, Indonesia. Materials and Methods: Lactic acid bacteria were grown on de Man–Ragosa–Sharpe agar medium for 24 hours. The bac- terial isolate with the best inhibitory power was identified as Bacillus subtilis (B. subtilis), based on 16S RNA sequences. Antimicrobial activity of the selected lactic acid bacteria was tested on the pathogenic bacteria, E. coli and Staphylococcus aureus. Using well diffusion method. In this study, in silico study was conducted to examine the structure and binding affin- ity of lactic acid bacteria against E. coli and S. aureus. Molecular docking experiments were performed using the PyRx 0.8 software. Results: This study showed that the bacteria were B. subtilis strain PATA-5. The response of inhibition of antimicrobial compounds produced by B. subtilis strain PATA-5 maximum in the stationary phase. The bactericidal properties of B. subtilis strain PATA-5 were categorized as strong against Gram-negative E. coli, i.e., 30.5 mm, when compared to Gram-positive S. aureus, i.e., 17.5 mm. Conclusion: B. subtilis strain PATA-5 is capable to produce natural antibiotic cyclic lipopeptides, namely surfactin.

Download Full-text

In vitro adhesion of lactic acid bacteria and bifidobacteria to Caco-2P and IEC-18 cells

Acta Alimentaria ◽

10.1556/aalim.34.2005.1.12 ◽

2005 ◽

Vol 34 (1) ◽

pp. 91-99 ◽

Cited By ~ 5

Author(s):

K. Szekér ◽

J. Beczner ◽

A. Halász ◽

Á. Mayer ◽

J.M. Rezessy-Szabó ◽

...

Keyword(s):

Lactic Acid ◽

Lactic Acid Bacteria ◽

In Vitro Adhesion

Download Full-text

Improving ensiling characteristics by adding lactic acid bacteria modifies in vitro digestibility and methane production of forage-sorghum mixture silage

Scientific Reports ◽

10.1038/s41598-021-81505-z ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Chatchai Kaewpila ◽

Pongsatorn Gunun ◽

Piyawit Kesorn ◽

Sayan Subepang ◽

Suwit Thip-uten ◽

...

Keyword(s):

Lactic Acid ◽

Lactic Acid Bacteria ◽

Nutritive Value ◽

Gas Production ◽

In Vitro Digestibility ◽

Acid Fermentation ◽

Design Factor ◽

Forage Sorghum ◽

Randomized Design

AbstractImproving the nutrition of livestock is an important aspect of global food production sustainability. This study verified whether lactic acid bacteria (LAB) inoculant could promote ensiling characteristics, nutritive value, and in vitro enteric methane (CH4) mitigation of forage sorghum (FS) mixture silage in attacking malnutrition in Zebu beef cattle. The FS at the soft dough stage, Cavalcade hay (CH), and cassava chip (CC) were obtained. The treatments were designed as a 4 × 2 factorial arrangement in a completely randomized design. Factor A was FS prepared without or with CH, CC, and CH + CC. Factor B was untreated or treated with Lactobacillus casei TH14. The results showed that all FS mixture silages preserved well with lower pH values below 4.0 and higher lactic acid contents above 56.4 g/kg dry matter (DM). Adding LAB boosted the lactic acid content of silages. After 24 h and 48 h of in vitro rumen incubation, the CC-treated silage increased in vitro DM digestibility (IVDMD) with increased total gas production and CH4 production. The LAB-treated silage increased IVDMD but decreased CH4 production. Thus, the addition of L. casei TH14 inoculant could improve lactic acid fermentation, in vitro digestibility, and CH4 mitigation in the FS mixture silages.

Download Full-text

Co-Microencapsulated Black Rice Anthocyanins and Lactic Acid Bacteria: Evidence on Powders Profile and In Vitro Digestion

Molecules ◽

10.3390/molecules26092579 ◽

2021 ◽

Vol 26 (9) ◽

pp. 2579

Author(s):

Carmen-Alina Bolea ◽

Mihaela Cotârleț ◽

Elena Enachi ◽

Vasilica Barbu ◽

Nicoleta Stănciuc

Keyword(s):

Lactic Acid ◽

Lactic Acid Bacteria ◽

Oryza Sativa L ◽

Milk Proteins ◽

Ethanolic Extract ◽

Dry Weight ◽

Hot Water ◽

Raw Material ◽

Black Rice

Two multi-functional powders, in terms of anthocyanins from black rice (Oryza sativa L.) and lactic acid bacteria (Lactobacillus paracasei, L. casei 431®) were obtained through co-microencapsulation into a biopolymer matrix composed of milk proteins and inulin. Two extracts were obtained using black rice flour as a raw material and hot water and ethanol as solvents. Both powders (called P1 for aqueous extract and P2 for ethanolic extract) proved to be rich sources of valuable bioactives, with microencapsulation efficiency up to 80%, both for anthocyanins and lactic acid bacteria. A higher content of anthocyanins was found in P1, of 102.91 ± 1.83 mg cyanindin-3-O-glucoside (C3G)/g dry weight (DW) when compared with only 27.60 ± 17.36 mg C3G/g DW in P2. The morphological analysis revealed the presence of large, thin, and fragile structures, with different sizes. A different pattern of gastric digestion was observed, with a highly protective effect of the matrix in P1 and a maximum decrease in anthocyanins of approximatively 44% in P2. In intestinal juice, the anthocyanins decreased significantly in P2, reaching a maximum of 97% at the end of digestion; whereas in P1, more than 45% from the initial anthocyanins content remained in the microparticles. Overall, the short-term storage stability test revealed a release of bioactive from P2 and a decrease in P1. The viable cells of lactic acid bacteria after 21 days of storage reached 7 log colony forming units (CFU)/g DW.

Download Full-text

Effect of lactic acid bacteria inoculants on alfalfa (Medicago sativa L.) silage quality: assessment of degradation (in situ) and gas production (in vitro)

Journal of Integrative Agriculture ◽

10.1016/s2095-3119(16)61424-7 ◽

2016 ◽

Vol 15 (12) ◽

pp. 2834-2841 ◽

Cited By ~ 7

Author(s):

Ce LIU ◽

Yu-jiao LAI ◽

Xiao-nan LU ◽

Ping-ting GUO ◽

Hai-ling LUO

Keyword(s):

Lactic Acid ◽

Lactic Acid Bacteria ◽

Quality Assessment ◽

Medicago Sativa ◽

Gas Production ◽

Medicago Sativa L ◽

Silage Quality

Download Full-text

Probiotic Potential of a Novel Vitamin B2-Overproducing Lactobacillus plantarum Strain, HY7715, Isolated from Kimchi

Applied Sciences ◽

10.3390/app11135765 ◽

2021 ◽

Vol 11 (13) ◽

pp. 5765

Author(s):

Joo-Yun Kim ◽

Eun-Jung Choi ◽

Jae-Ho Lee ◽

Myeong-Seok Yoo ◽

Keon Heo ◽

...

Keyword(s):

Lactic Acid ◽

Lactic Acid Bacteria ◽

Lactobacillus Plantarum ◽

Probiotic Strain ◽

Optimal Growth ◽

Control Group ◽

High Survival ◽

Vitamin B2 ◽

Riboflavin Deficiency

Vitamin B2, also known as riboflavin, is essential for maintaining human health. The purpose of this study was to isolate novel lactic acid bacteria that overproduce vitamin B2 and to validate their potential as probiotics. In this study, Lactobacillus plantarum HY7715 (HY7715) was selected among lactic acid bacteria isolated from Kimchi. HY7715 showed a very high riboflavin-producing ability compared to the control strain due to the high expression of ribA, ribB, ribC, ribH, and ribG genes. HY7715 produced 34.5 ± 2.41 mg/L of riboflavin for 24 h without consuming riboflavin in the medium under optimal growth conditions. It was able to produce riboflavin in an in vitro model of the intestinal environment. In addition, when riboflavin deficiency was induced in mice through nutritional restriction, higher levels of riboflavin were detected in plasma and urine in the HY7715 administration group than in the control group. HY7715 showed high survival rate in simulated gastrointestinal conditions and had antibiotic resistance below the cutoff MIC value suggested by the European Food Safety Authority; moreover, it did not cause hemolysis. In conclusion, HY7715 could be considered a beneficial probiotic strain for human and animal applications, suggesting that it could be a new alternative to address riboflavin deficiency.

Download Full-text

Isolation of lactic acid bacteria from swine milk and characterization of potential probiotic strains with antagonistic effects against swine-associated gastrointestinal pathogens

Canadian Journal of Microbiology ◽

10.1139/cjm-2015-0811 ◽

2016 ◽

Vol 62 (6) ◽

pp. 514-524 ◽

Cited By ~ 8

Author(s):

Sandra Rayén Quilodrán-Vega ◽

Julio Villena ◽

José Valdebenito ◽

María José Salas ◽

Cristian Parra ◽

...

Keyword(s):

Lactic Acid ◽

Lactic Acid Bacteria ◽

Human Milk ◽

Salmonella Enterica ◽

Enterohemorrhagic Escherichia Coli ◽

Salmonella Infection ◽

Antagonistic Effects ◽

Gastrointestinal Pathogens

Probiotics are usually isolated from the gastrointestinal tract of humans and animals. The search of probiotics in human milk is a recent field of research, as the existence of the human milk microbiome was discovered only about a decade ago. To our knowledge, no reports regarding the potential probiotic effect of bacteria from swine milk have been published. In this work, we isolated several lactic acid bacteria from swine milk and evaluated them for them potential as probiotics. Among the isolated strains, Lactobacillus curvatus TUCO-5E showed antagonistic effects against swine-associated gastrointestinal pathogens. TUCO-5E was able to reduce the growth of enterotoxigenic and enterohemorrhagic Escherichia coli strains as well as pathogenic salmonella. In vitro exclusion and displacement assays in intestinal epithelial cells showed a remarkable antagonistic effect for L. curvatus TUCO-5E against Salmonella sp. strain TUCO-I7 and Salmonella enterica ATCC 13096. Moreover, by using a mouse model of Salmonella infection, we were able to demonstrate that preventative administration of L. curvatus TUCO-5E for 5 consecutive days was capable of decreasing the number of Salmonella enterica serovar Typhimurium in the liver and spleen of treated mice, compared with the controls, and prevented dissemination of the pathogen to the blood stream. Therefore, we have demonstrated here that swine milk is an interesting source of beneficial bacteria. In addition, the results of this work suggest that L. curvatus TUCO-5E is a good candidate to study in vivo the protective effect of probiotics against intestinal infection and damage induced by Salmonella infection in the porcine host.

Download Full-text