Traditional Fermented Buffalo Milk Research Articles

Lactococcus lactis D4 Has Potential Effect to Alleviate Inflammation and Reverse Dysbiosis in Colitis Rat Model

BACKGROUND: Inflammatory bowel disease (IBD) is a prevalent chronic inflammatory disorder of the gastrointestinal tract that may lead to colorectal cancer if untreated. Several medications for IBD have adverse side effects. Thus, there are high interest for searching alternative treatment to treat IBD. Lactococcus lactis D4 isolated from dadih, a traditional fermented buffalo milk product, was investigated for the effect on gut inflammation and microbial composition in the colitis rat model.METHODS: Eighteen male Sprague-Dawley rats were divided into three groups; control rats, colitis-induced rats, and colitis-induced rats treated with L. lactis D4 isolate. The control group received water, while the colitis-induced rats were given azoxymethane (AOM) and dextran sodium sulphate (DSS). Rats’ feces were collected for the analysis of gut microbiota diversity by next-generation sequencing and for the measurement of transforming growth factor (TGF)-β, nuclear factor kappa-B (NF-κB), interleukin (IL)-6, and tumor necrosis factor (TNF)-α colonic expression using reverse transcription-quantitative polymerase chain reaction (RT-qPCR).RESULTS: L. lactis D4 administration was able to reduce inflammation in colitis by decreasing IL-6 expression (0.87 vs. 0.73), while expression of TGF-β, NF-κB, and TNF-α were increased compared to the model group, suggesting a complex immunomodulatory effect. Gut microbiota sequencing revealed a similarity between the control and treatment groups, suggesting L. lactis D4 has great potential to ameliorate gut microbiota dysbiosis in colitis rats.CONCLUSION: L. lactis D4 has a beneficial effect in decreasing pro-inflammatory cytokines and is able to reserve dysbiosis in colitis rat model. Thus, L. lactis D4 might be used as a therapeutic agent for IBD.KEYWORDS: colitis, gut microbiota, inflammation, inflammatory bowel disease, Lactococcus lactis D4, probiotic, proinflammatory cytokines

Ampiang-Dadih-a combination of Indonesian traditional fermented buffalo milk and black glutinous rice-prevents hypercholesterolemia and liver cell degeneration in vivo: A pilot study.

We aimed to evaluate the potential of Ampiang-Dadih (AD), a combination of Indonesian traditional fermented buffalo milk (Dadih) and black glutinous rice flakes (Ampiang) as an anti-hypercholesterolemic agent and to prevent liver-cell degeneration using a rat model. A mixture of black glutinous rice powder (0.3 gm/gm feed) and fermented buffalo milk (3.74/200 gm BW) was prepared to obtain AD. Fifteen adult male Sprague Dawley rats were divided into three groups of five animals each: (A) negative control group (distilled water; 5 weeks), (B) hypercholesterolemia group (1% cholesterol per feed; 5 weeks), and (C) preventive AD group (1% cholesterol and AD; 5 weeks). The blood lipid profiles were measured at weeks 2, 4, and 5. The liver enzyme activity, cholesterol level, and histology were observed at the end of week 5. AD administration simultaneously with cholesterol in Group C significantly prevented an increase in total plasma cholesterol and low-density lipoprotein levels compared to Group B. Alanine transaminase and aspartate transaminase were maintained at normal levels in Group C. Furthermore, the levels of liver cholesterol and liver cell degeneration in Group C were also maintained because of AD administration compared to that in Group B. This study demonstrated that AD has the potential to be developed as a functional food for hypercholesterolemia prevention.

I am better than I look: genome based safety assessment of the probiotic Lactiplantibacillus plantarum IS-10506

BackgroundSafety of probiotic strains that are used in human and animal trials is a prerequisite. Genome based safety assessment of probiotics has gained popularity due its cost efficiency and speed, and even became a part of national regulation on foods containing probiotics in Indonesia. However, reliability of the safety assessment based only on a full genome sequence is not clear. Here, for the first time, we sequenced, assembled, and analysed the genome of the probiotic strain Lactiplantibacillus plantarum IS-10506, that was isolated from dadih, a traditional fermented buffalo milk. The strain has already been used as a probiotic for more than a decade, and in several clinical trials proven to be completely safe.MethodsThe genome of the probiotic strain L. plantarum IS-10506 was sequenced using Nanopore sequencing technology, assembled, annotated and screened for potential harmful (PH) and beneficial genomic features. The presence of the PH features was assessed from general annotation, as well as with the use of specialised tools. In addition, PH regions in the genome were compared to all other probiotic and non-probiotic L. plantarum strains available in the NCBI RefSeq database.ResultsFor the first time, a high-quality complete genome of L. plantarum IS-10506 was obtained, and an extensive search for PH and a beneficial signature was performed. We discovered a number of PH features within the genome of L. plantarum IS-10506 based on the general annotation, including various antibiotic resistant genes (AMR); however, with a few exceptions, bioinformatics tools specifically developed for AMR detection did not confirm their presence. We further demonstrated the presence of the detected PH genes across multiple L. plantarum strains, including probiotics, and overall high genetic similarities between strains.ConclusionThe genome of L. plantarum IS-10506 is predicted to have several PH features. However, the strain has been utilized as a probiotic for over a decade in several clinical trials without any adverse effects, even in immunocompromised children with HIV infection and undernourished children. This implies the presence of PH feature signatures within the probiotic genome does not necessarily indicate their manifestation during administration. Importantly, specialized tools for the search of PH features were found more robust and should be preferred over manual searches in a general annotation.

Physicochemical and microbiological study of Dadiah jelly candy with the addition of Clitoria ternatea flower extract

Dadiah is a traditional fermented buffalo milk originating from West Sumatra, Indonesia and has health benefits. The aim of this study was to evaluate the effect of the addition of Clitoria ternatea flower extract addition (0,5%, 1%, 1,5%, and 2%) on the moisture content, hardness, total lactic acid bacteria, antioxidant activity, and total phenolic content of dadiah jelly candies. The results showed that the addition of Clitoria ternatea extract increased the moisture content, decreased the hardness value, increased the total phenolic content, and enhanced the antioxidant activity of the jelly candies. Compared to the dadiah jelly candies without Clitoria ternatea addition, jelly candies with Clitoria ternatea addition at any concentration did not differ significantly (P > 0.05) in total lactic acid bacteria, whereas the pH value increased as the concentration of the Clitoria ternatea extract increased. Overall, the use of Clitoria ternatea flower extract can improve the functional properties of dadiah jelly candies, with the best concentration of 2% of the Clitoria ternatea extract on the antioxidant activity and total phenol content.

Immune Response and Nutritional Status of Infants’ Birth after Supplementation with Dadih and Zinc during Pregnancy: A Prospective Intervention Study

BACKGROUND: Dadih is an Indonesian traditional fermented buffalo milk produced and consumed by the secretory immunoglobulin A (sIgA) level and nutritional status of infants aged 4–10 months. AIM: This study aims to assess the effect of dadih and zinc supplementation given to pregnant mothers for 6 months on fecal. MATERIALS AND METHODS: A prospective randomized controlled trial was conducted in two districts on 138 pregnant mothers for 6 months, randomly assigned into three groups, namely, control, dadih, combination of dadih and zinc groups. Then, the combination of groups was supplemented to expecting mothers, respectively, 6 times a week during 6 months. Anthropometric measurement and fecal sIgA of infants were assessed. RESULTS: The prevalence of nutritional status of infants who mothers were supplemented during pregnancy with dadih, dadih and zinc, and, normal diet, respectively, are 10% stunting, 5.0% underweight, and 5.0% wasting infants; and 22.5% stunted, 20.0% underweight, and 5.0% wasting infants were found in expecting mothers supplemented with it and without any supplementation (control group). The percentage of stunting in 8 months and older infants tended to reduce: 28.57%, 31.25%, and 11.76%. Their mothers were supplemented with all groups. CONCLUSION: Dadih and zinc are potential for improving nutritional status of the newborn. In addition, the two types of supplementation are recommended to pregnant mothers, to reduce the prevalence of stunting. Potential probiotic contained in Dadih which consumed in adequate amounts can help the absorption of nutrients and increase immunity of mothers and infants amounts confer a health benefit. Acceleration and growth of infants to achieve optimal growth and development should be carried out before they are under two years old.

Whole-genome sequence analysis and probiotic characteristics of Lactococcus lactis Subsp. lactis strain Lac3 isolated from traditional fermented buffalo milk (Dadih)

BackgroundProbiotics are live microorganisms that provide beneficial effects on the host’s health when exploited in adequate amounts. This study aimed at carrying out whole-genome sequence analysis and in vitro potential probiotic characteristics of Lactococcus lactis subsp. lactis strain Lac3 isolated from the spontaneously fermented buffalo milk named Dadih. ResultsThe results from de novo assembly indicated that the assembled genome consisted of 55 contigs with a genome size of 2,441,808 bp ~ (2.44 Mb), and GC % content of 34.85%. The evolution history result showed that the strain Lac3 was closely related to Lactococcus lactis species deposited in NCBI with a sequence similarity ≥ 99.93%. L. lactis subsp. lactis Lac3 was non-pathogenic with a probability of 0.21 out of 1 and had a pathogenicity score of zero (0), and neither harbored virulence factors nor acquired antibiotic resistance phenotypes. L. lactis subsp. lactis Lac3 exhibited the potential probiotic characteristics to tolerate acid at pH (2.0 and 5.0), salinity (1–5% NaCl), bile salt of (0.3–1.0%) and had auto-aggregation capacity increased from 6.0 to 13.1%. ConclusionThis study described a novel strain of Lactococcus lactis subsp. lactis called Lac3, which exhibits probiotic properties that could be beneficial in the development of probiotics.

Purification and Partial Characterization of a Bacteriocin Produced by Lactobacillus pentosus 124-2 Isolated from Dadih

The fermentation process results in the presence of lactic acid bacteria (LAB). Traditional fermented buffalo milk from the Indonesian province of West Sumatra is known as dadih. Bacteriocins are naturally occurring antimicrobial peptides that can be produced by LAB. These bacteriocins have the potential to be used as natural preservatives. This research was conducted with the intention of isolating and partially characterizing a bacteriocin that was generated by Lactobacillus pentosus 124-2 that was isolated from dadih. Using MRS agar media and 16sRNA, the LAB that was found in dadih was successfully isolated. Bacteriocins were produced using media consisting of MRS broth. After the bacteriocins were obtained, they were subjected to a series of purification processes, one of which included ammonium sulfate (70%) followed by gel filtration chromatography for additional purification. According to the findings, the strain of LAB that was discovered in the dadih was Lactobacillus pentosus 124-2. The specific activity of the bacteriocins rose, allowing for the effective completion of the purification process. The pure bacteriocins had the greatest specific activity values, at 60.59 AU/mg, while the yield values were 0.41% and 3.15-fold. Bacteriocins in their purest form were able to inhibit the growth of Salmonella species as well as Staphylococcus aureus. The characterization results included partial bacteriocins that were resistant to pH 2, 4, and 6; antibacterial activity that was was stable at a temperature range of 25 °C to 121 °C; and resistance to the addition of 2%, 4%, and 6% salt concentrations. Microbial stability against the addition of surfactants EDTA, SDS, and Tween 80 was also obtained. In light of these findings, a bacteriocin derived from L. pentosus possesses the possible ability to be utilized in the food business as a biopreservative.

The Potential Lactic Acid Bacteria from Dadiah Sianok Bukittinggi City, West Sumatera as Probiotic

Dadiah is a traditional fermented buffalo milk from West Sumatra, Indonesia. It is one of the healthiest drinks because it contains lactic acid bacteria (LAB), which has many health benefits. Lactic Acid Bacteria (LAB) are a group of bacteria that play a role in the fermentation process of food. Lactic Acid Bacteria (LAB) content in dadiah will affect the quality of dadiah in general.West Sumatra has several dadiah producing areas, one of which is the Sianok area located in the city of Bukittinggi. This research aims to find out the potential of Sianokdadiah as a probiotic food. The method used in this research is a survey method with descriptive analysis. Lactic Acid Bacteria (LAB) contained in dadiah was isolated using de Man RogosaSharge (MRS) media. The isolates were then identified based on their morphology and biochemical properties. The tests carried out include gramme stain, catalase test, fermentative type, retention of gastric juice and bile salts. Furthermore, testing using 16S rRNA molecular identification techniques was conducted to determine the species level. The results of the study obtained a total colony of Lactic Acid Bacteria (LAB) from dadiah 89x109 CFU/g. The identification of the single colony found that the Lactic Acid Bacteria (LAB) obtained was included as Gram-positive, with the type of homofermentative fermentation, catalase negative. It has resistance to stomach acid at pH 3 with a viability of 83.7%, bile salt resistance of 0.3%, and viability of 67.3%. Identification of Lactic Acid Bacteria (LAB) using the 16S sRNA gene, the results of running PCR with a base length of 1428bp. Analysis based on phylogenetic trees showed that Lactic Acid Bacteria (LAB)dadiah from the city of SianokBukittinggi has a relationship with Pediococcusacidilactici. From the results of this research, it can be concluded that dadiah from the city of Sianok Bukittinggi has a good enough potential as a probiotic.

Evaluation of the Probiotic Potential of Lactobacillus delbrueckii ssp. indicus WDS-7 Isolated from Chinese Traditional Fermented Buffalo Milk In Vitro.

The present study aimed to evaluate the probiotic potential of lactic acid bacteria (LAB) isolated from Chinese traditional fermented buffalo milk. Out of 22 isolates, 11 were putatively identified as LAB preliminarily. A total of six LAB strains displayed strong adhesion to HT-29 cells and all these strains showed preferable tolerance to artificially simulated gastrointestinal juices. WDS-4, WDS-7, and WDS-18 exhibited excellent antioxidant capacities, including DPPH radical, ABTS+ radical, and superoxide anion scavenging activities. Compared with the other two LAB strains, WDS-7 had a stronger inhibition effect on four pathogens. Based on the 16S rRNA gene sequencing and phylogenetic analysis, WDS-7 was identified as Lactobacillus delbrueckii ssp. indicus and selected to assess the potential and safety of probiotics further. The results revealed that WDS-7 strain had a strong capacity for acid production and good thermal stability. WDS-7 strain also possessed bile salt hydrolase (BSH) activity. Compared to LGG, WDS-7 was a greater biofilm producer on the plastic surface and exhibited a better EPS production ability (1.94 mg/ml as a glucose equivalent). WDS-7 was proved to be sensitive in the majority of tested antibiotics and absence of hemolytic activity. Moreover, no production of biogenic amines and β-glucuronidase was observed in WDS-7. The findings of this work indicated that L. delbrueckii ssp. indicus WDS-7 fulfilled the probiotic criteria in vitro and could be exploited for further evaluation in vivo.

Antimicrobial activity of bacteriocin like inhibitory substance (BLIS) and lactic acid bacteria (LAB) isolated from traditional fermented buffalo milk from West Sumatra, Indonesia

Food preservation by chemical preservatives addition is not recommended in large quantities. However, lactic acid bacteria (LAB) metabolites can be used as a safe alternative. LAB can produce a type of antimicrobial peptide known as bacteriocin. One source of LAB that can be used as a bacteriocin-producing candidate is dadih, a traditional fermented buffalo milk product from West Sumatra, Indonesia. In this study, eight bacterial isolates isolated from dadih were identified using the catalase test and Gram straining and obtained 6 isolates that were taken as LAB. An antimicrobial activity test was carried out using the diffusion method on LAB liquid isolates suspension and crude bacteriocins from the six isolates. The isolates with the highest antimicrobial activity were DK8, the diameter of inhibiting zone of liquid isolates was 15.75 mm (Salmonella sp.), 14.25 mm (Escherichia coli), and 12.25 mm (Staphylococcus aureus). Meanwhile, the diameter of crude bacteriocin inhibition of DK 8 isolates was 8 mm (Salmonella sp.). 8 mm (E. coli), and 8.5 mm (S. aureus). Furthermore, 16S rRNA sequencing confirmed DK8 as Lactobacillus pentosus.

Isolation, identification and characterization of Lactobacillus species diversity from Meekiri: traditional fermented buffalo milk gels in Sri Lanka

Traditional fermented buffalo milk gel; Meekiri, is a popular buffalo milk-derived product in Sri Lanka. Predominantly, it is produced using the back-slopping (adding a small amount of the previous fermentate) technique, following the life-long traditions available at the cottage level. Hence, diverse and unclassified starter cultures are likely to be established across the varying geographical regions of Meekiri production. In the present study, we aimed to elucidate the diversity of lactic acid bacteria (LAB) and their characteristics including probiotic properties from major Meekeri production areas (n = 22) in Sri Lanka. Lactic acid bacteria was isolated from locally produced Meekiri samples (n = 23) and characterized based on morphological, biochemical, physiological profiles and potential of probiotic properties. The isolates revealed five different colony and cell morphologies and were classified as heterofermenters, homofermenters and facultative heterofermenters based on CO2 production using glucose. None of the isolates showed the ability to grow either at 5 °C or 0 °C, while 71 % and 100 % survival of the isolates were observed at 15 °C and 45 °C, respectively. Amplified ribosomal DNA restriction analysis (ARDRA) primarily grouped the isolates into three distinct clusters based on their DNA banding patterns. Subsequently, 16S rRNA sequencing of isolates revealed the presence of four species namely, Limosilactobacillus fermentum (n = 18), Latilactobacillus curvatus (n = 2), Lactobacillus acidophilus (n = 2) and Lactiplantibacillus plantarum (n = 1) and in the phylogenetic analysis, it was represented by four distinctive clades. All the isolated species demonstrated promising probiotic potential with antibiotic sensitivity, antimicrobial properties, bile acid tolerance and acid tolerance. In conclusion, traditional back-slopping Meekiri in Sri Lanka contains diverse LAB, with a negligible geographical variation at species-level. Our work provides a strong foundation and insights into future applications in starter culture development for the fermentation of buffalo's milk.

Comparative lactic acid bacteria (LAB) profiles during dadih fermentation with spontaneous and back-slopping methods, as identified by terminal-restriction fragment length polymorphism (T-RFLP)

The diversity of lactic acid bacteria (LAB) present during the manufacture of traditional fermented buffalo milk from West Sumatra, known as dadih, was studied via a culture-independent approach using terminal-restriction fragment length polymorphism (T-RFLP) to compare the dynamic diversity in back-slopping and spontaneous fermentation methods. Total LAB and pH were measured in freshly prepared buffalo milk and in \textit{dadih} fermented for 24 and 48 hours. The results indicated significant differences between the fermentation methods, with higher total LAB, and greater phylotype richness and relative abundance being identified in the back-slopping method. Terminal fragment lengths (TRFs) of 68 and 310 bp were common to both techniques, similar to those of Lactobacillus fermentum, Fructobacillus pseudoficulneus, Leuconostoc citreum, Leuconostoc kimchii, and Leuconostoc sp. The changes in phylotype number (species number) and relative abundances of LAB communities identified are expected to produce data needed to formulate the best fermentation process for dadih manufacturing. A 24-hour back-slopping fermentation method is recommended, as fermentation time of longer than 24 hours reduced viable LAB significantly. Our results also indicated that the T-RFLP technique is not only clearly sensitive enough and adequate for segregating LAB diversity in both fermentation methods, but that it also provides good information regarding the structure of microbial communities and their composition change during the fermentation process.

Characterization of Dadih: Traditional Fermented Buffalo Milk of Minangkabau

Dadih or dadiah is traditional fermented buffalo milk of Minangkabau, which occurs spontaneously. Dadih is commonly served as ampiang dadih, or other dishes. The microbiota found in dadih are dominated by lactic acid bacteria, and yeasts are also found. The lactic acid bacteria provide functional value, such as antimicrobial, hypocholesterolemic, antimutagenic, antioxidant, and immunomodulatory properties, as well as being the source of γ-aminobutyric acid (GABA) as an anti-stress agent and folate. Nevertheless, many challenges were observed in dadih production, including the limitation of buffalo milk production due to decreasing populations of buffalo in the last two decades, unstandardized dadih production due to the spontaneous fermentation in natural bamboo tubes, and safety problems as no heat treatment is applied in the production of dadih. These problems impede the development of dadih production, thus is it important to improve buffalo cultivation through artificial insemination programs, using different types of milk and pasteurization processes in dadih production, and incubator development to accelerate the fermentation period.

Survival of Lactobacillus plantarum dad 13 in probiotic cheese making

Lactobacillus plantarum Dad 13 is a group of Lactic Acid Bacteria, which was isolated from “Dadih” (traditional fermented buffalo milk) has known as indigenous probiotic from Indonesia. Probiotics are defined as living microorganisms and have health benefits with consumption for use as a supplement of food with the amount of cell viability at least 108 cells. L. plantarum frequently used as a starter in probiotic drink, especially in fermented milk. Milk is a valuable source of nutritional substances with the composition of water, protein, fat, sugars, mineral salts, vitamins, and enzymes for the living of microorganisms. To study the potential of L. plantarum Dad 13 in milk, we present an updated inventory of L. plantarum Dad 13 used in milk to making cheese. These we are applied L. plantarum Dad 13 to produce lactic acid for making curds. Combination treatment of biomass production used for cheese making that was biomass production using coconut water and MRS medium. The different combinations of a medium can influence the biomass viability of L. plantarum Dad 13 in cheese. The result showed that the viability of L. plantarum Dad 13 in cheese using two kinds of media during the production of biomass (i.e., coconut water and MRS) were almost similar during two months of storage, that was 103 cfu/g. They decreased on viability after two-month storage was about 3 log cycles. The result showed that viable the cell of L. plantarum Dad 13 in this cheese did not agree with the criterion of a minimum of 106 – 108 cfu/g viable cells as a probiotic product. Overall, local isolates of L. plantarum Dad 13 can be applied in the process of cheese making.

Half-life Estimation of Encapsulated Enterococcus faecium IS-27526 by Accelerated Shelf Life Testing (ASLT)

Probiotics are live microbes with beneficial effect to the host when administrated in adequate quantities. Ensuring the functionality of probiotics can be a challenge for manufacturers due to harsh environment conditions during processing, handling and storage. Enterococcus faecium IS-27526 is one of novel probiotic strain isolated from dadih, traditional fermented buffalo milk of West Sumatra. The purpose of this study was to determine the effect of microencapsulation, moisture absorber, and storage temperature on the half-life of probiotic powder estimated by Accelerated Shelf Life Testing (ASLT). There were two treatment groups, namely free cell (FC), and microencapsulated cell (EC), with additional treatment in each group, with addition of moisture absorber (MA) and without moisture absorber, stored at various temperatures (5°C, 27°C and 37°C) for 21 days and viable counts was assessed every 7 days. Storage temperature significantly affects the half-life of probiotic powder (p<0.05). At 5°C, EC significantly showed longer half-life than EC-MA, 140.14 ± 5.701 and 113.45 ± 3.242 days, respectively, at 27°C, EC significantly shorter half-life than EC-MA, 38.64 ± 6.831 and 42.35 ± 0.973 days, respectively. Microencapsulation and moisture absorber didn’t show significant effect on half-life (p>0.05) at various temperature storage.

Preparation of Indigenous Lactic Acid Bacteria Starter Cultures for Large Scale Production of Fermented Milk

&lt;div&gt;Lactobacillus plantarum Dad 13, an indigenous probiotic was examined its ability to be used as a single starter culture or mixed cultures with Streptococcus thermophilus Dad 11 for milk fermentation. Both cultures were isolated from dadih, a traditional fermented buffalo milk. The purposes of this study were to produce indigenous lactic acid bacteria starter cultures using halal growth medium and evaluate their application on large scale fermented milk production. The halal medium was developed using natural compounds such as sucrose, meat peptone, mung bean sprout extract, tomato extract, and young coconut water. Meat peptone was prepared by hydrolysis of halal meat using crude bromelain. Lactic acid bacteria were grown in the halal growth medium then harvested, frozen and freeze-dried. A single freeze-dried starter culture of L. plantarum Dad 13 and frozen mixed cultures of L. plantarum Dad 13 and S. thermophilus Dad 11 were prepared for production of fermented milk drink and yogurt respectively in industrial scale. The growth of these lactic acid bacteria in halal growth medium increased the viable cell to two log cycles (109 CFU/mL) for L. plantarum Dad 13 and one log cycle for S. thermophilus Dad 11 (108 CFU/mL), respectively. The viable cell of freeze-dried L. plantarum Dad 13 and S. thermophilus Dad 11 were 7.57 x 1010 CFU/g and 6.35 x 109 CFU/g, respectively. The number of viable cells and pH of both fermented milk drink and yogurt products was relatively stable to 107 CFU/mL and 108 CFU/mL, respectively during cold storage for four to six weeks. The sensory characteristics of the products were comparable to the ones using commercial starter cultures. It can be concluded that these indigenous starter cultures can be applied for the production of probiotic fermented milk.&lt;/div&gt;

Characteristics of Amino Acid, Micronutrient and Probiotic Isolated from Dadih and Their Benefits for Pregnant Mothers and Outcomes in West Sumatra, Indonesia

Dadih is a traditional fermented buffalo milk consumed by the Minangkabau people in West Sumatera Province, Indonesia, especially in Bukittinggi, Padang Panjang, Solok, Lima Puluh Kota, and Tanah Datar districts. The objective of this study was to identify amino acid and strain of acid lactic bacteria isolated from dadih and to evaluate the benefit of dadih intervention during pregnancy on the nutritional status of an infant&amp;#39;s birth. This study was initiated with DNA extraction from bacterial cultures, Ribosomal DNA amplification, and sequencing, analysis of amino acid using UPLC. The intervention was conducted in two districts on 138 pregnant mothers for 6 months. A prospective cohort study was conducted on a follow up of a previous randomized controlled trial. This study was conducted in two districts in West Sumatera Province from April 2018 to July 2019. Spontaneous probiotic species of dadih strain were identified in this study: Lactobacillus Plantarum, Lactobacillus casei, Lactobacillus durans, Leuconostoc pseudo mesentoroides, and Lactobacillus cactis. Glutamic acid was the highest quantity of non-essential amino acid in dadih (16.28 mg/g), while Lysine and Leucine were the highest essential amino acids (7.22 and 6.42 mg/g). The study revealed that after a 6-month intervention, about 66.7% of infants birth weight &amp;gt; 3000 grams and only 2.1% of infants having birth weight &amp;lt; 2500 grams for Dadih Group and about 60.0% had infant birth weight &amp;gt; 3000 grams and 4.4% infants with low birth in Control Group. The result of this study shows that there are more benefits in pregnant mothers&amp;#39; weight gain and infants&amp;#39; birth weight until a six-month follow up from pregnant mothers in Dadih Group than Control Group. The results of the follow-up study show that infants from mothers supplemented with Dadih Zinc Group have significantly less undernutrition status as compared to the Control Group (p&amp;lt;0,05). The results also show that dadih has a sufficient amount of protein to fulfill the nutritional need for pregnant mothers and the outcomes.

Lactobacillus plantarum IS-10506 promotes renal tubular regeneration in pyelonephritic rats.

Lactobacillus plantarum IS-10506 is a novel probiotic isolated from Indonesian traditional fermented buffalo milk dadih. Probiotics are clinically proven to be effective in accelerating recovery after urinary tract infection (UTI) and in decreasing recurrent UTI in children with or without structural disorders of the urinary tract. This study aimed to investigate the role of the probiotic L. plantarum IS-10506 in activating and regenerating renal epithelial stem cells in pyelonephritic rats. Fifty-five 20-weeks-old male Sprague-Dawley rats were randomised into control, lipopolysaccharide (LPS) and treatment (LPS and probiotic) groups. Pyelonephritis was induced with Escherichia coli ATCC 25922 LPS administered via a urethral catheter on the first experimental day to the LPS and treatment groups. Microencapsulated L. plantarum IS-10506 was orally administered once daily for up to 14 days to the treatment group. On days 3, 5, 7, 10 and 14, five rats per group were sacrificed and their kidneys were immunohistochemically analysed for production of interleukin (IL)-10, lipocalin-2 and Ki-67 in the renal tubular cells. IL-10 production was upregulated starting from day 3 through day 14 in the treatment group (P<0.05) compared with that in the control and LPS groups. Moreover, treatment with the probiotic led to increased activation of renal tubular stem cell, as indicated by a higher lipocalin-2 production, and further proliferation of renal tubular stem cells was documented by a higher Ki-67 production. Taken together, these results indicate that the anti-inflammatory probiotic L. plantarum IS-10506 improves renal injury in pyelonephritic rats by activating endogenous renal tubular stem cells to proliferate into mature renal tubular epithelial cells.

Effect of Lactobacillus casei subsp. casei R-68 Isolated from Dadih on the Procarcinogenic Enzyme Activity and Fecal Microflora Count of Rats Challenged with Pathogenic Bacteria

The human digestive tract is a complex ecosystem that may contain bacteria, yeast, and other microflora, which have harmful and beneficial effects on the host. Species of Lactobacillus and Bifidobacterium are most commonly used as probiotics. Lactobacillus casei subsp. casei R-68 (LCR-68) isolated from dadih, traditional fermented buffalo milk from West Sumatera has the potential to be used as probiotic. The purposes of the present study were to evaluate the ability of strain LCR68 to inhibit the growth of the pathogenic bacteria Listeria monocytogenes FNCC-0156 and Escherichia coli FNCC-19 and reduce the activity of fecal mutagen enzymes in Wistar rats. The in vivo test used 25 male Wistar rats with an average weight of 174 - 176 g. This study consisted of five groups of treatment with five rats of each group. The results show a significant increase in the growth in all groups, although a significantly lower weight gain was observed in rats challenged with Listeria monocytogenes and fed fermented milk LCR-68. The counts of aerobic and anaerobic microbes were the same in all groups. Significantly higher counts of lactic acid bacteria were determined after the application of fermented milk LCR68. Significantly lower counts of Escherichia coli were also observed after the application of fermented milk LCR68. The presence of LCR-68 in fermented milk reduced the activity of β-glucuronidase and β-glucosidase significantly in the feces of Wistar rats. Therefore, the strain R-68 as a probiotic is expected to be able to prevent the formation of procarcinogenic compounds into carcinogens that cause cancer in the digestive tract.

Nutritional quality of milk curd by adding avocado extract (Persea americana, mill) using gombong bamboo (Gigantochloa verticilata)

Curd is a traditional fermented buffalo milk from West Sumatra.Fermentation in curd making occurs spontaneously, meaning thatfermentation occurs naturally without the addition of a starter. This studyaims to determine the nutritional content of cow milk curd with the additionof avocado extract (Persea americana, Mill) various levels using Gombongbamboo (Gigantochloa verticilata).This research was conducted at the Analogy and Physiology Laboratory ofthe Faculty of Agriculture, University of North Sumatra in November 2018-December 2018. The design used in this study was a completelyrandomized design (CRD) with 5 treatments and 4 replications. Theparameters analyzed were Protein, Fat, and pH. The results showed thatcow milk curd with the addition of avocado (Persea americana, Mill)extract various levels using Gombong bamboo (Gigantochloa verticilata)significantly had different effects on the nutritional value of protein, fat andpH. From the nutritional value studied, the highest protein: P1 7.57% withthe addition of avocado fruit extract (Persea americana, Mill) 6%, fataverage: P1 7.08% with the addition of avocado fruit extract (Perseaamericana, Mill) 6%, pH: P4 5.30% with the addition of avocado fruitextract (Persea americana, Mill) 24%.