Lactic Acid Bacteria Research Articles

Screening appropriate lactic acid bacteria as adjunct starter to enhance characteristic flavor and sensory profiles of Huangjiu (Chinese rice wine)

Lactic acid bacteria (LAB) are crucial for enhancing Huangjiu flavor, yet no standardized criteria exist for selecting LAB strains for this purpose. This study aimed to investigate the suitability of screening appropriate LAB strains as adjunct starters for Huangjiu fermentation. Twenty-five LAB strains were assessed for their stress resistance and fermentation characteristics. Co-fermentation with Saccharomyces cerevisiae was conducted to explore its role in flavor development, as this yeast greatly contribute to desirable flavor profiles. Notably, Lactiplantibacillus plantarum AR1018, AR1026 and Weissella confusa AR1038 significantly reduced 3-methyl-1-butanol production while increasing levels of ethyl hexanoate and ethyl octanoate. These strains were selected for further analysis of their impact on the physicochemical properties, volatile organic compounds, and sensory attributes of Huangjiu. The LAB starters enhanced total acidity, sugar and protein utilization and ester synthesis, while reducing ethanol and alcohol levels. AR1018 achieved a 16.51% reduction in 3-methyl-1-butanol, whereas AR1038 exhibited a 27.32% increase in ester content compared to the control. Principal component analysis revealed distinct flavor characteristic between Huangjiu produced with AR1018 and AR1038, and sensory evaluation confirmed enhanced desirable attributes, especially for AR1038. This study highlights the potential of LAB starters in improving the flavor quality of rice wine and similar fermented beverages.

Comparative Effects of Crude Extracts and Bioactive Compounds from Bidens pilosa and Bidens alba on Nonspecific Immune Responses and Antibacterial Activity Against Vibrio sp. in Coculture with Lactic Acid Bacteria in Hybrid Grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂).

This study investigated the effect of substances on nonspecific immune responses of head kidney leukocytes, the antimicrobial activity against Vibrio sp., as well as the time-kill of Vibrio sp. by combining the substances with lactic acid bacteria (LAB) and Pediococcus sp. The substances are B. pilosa hot water extract, B. pilosa powder extract, B. pilosa methanol extract, B. pilosa ethanol extract, B. alba hot water extract, B. alba powder extract, B. alba methanol extract, B. alba ethanol extract, and bioactive compounds, namely cytopiloyne, flavonoid, phenol, ethyl caffeate, luteolin, chlorogenic acid, butein, and linoleic acid. The results showed that some of them were nontoxic to the head kidney leukocytes, which can increase the phagocytic rate, phagocytic index, and respiratory burst. These substances were able to inhibit the growth of Vibrio sp.; they can even completely kill the pathogenic bacteria. The largest of the inhibition zone formed from the EC group at a concentration range of 5-50 µg/mL against V. parahaemolyticus, V. alginolyticus, and V. harveyi with a value of 19.7 ± 0.56, 19.3 ± 1.53, and 20.6 ± 1.53 mm. Furthermore, the time-kill studies showed that the LAB and P. acidilactici can completely kill the Vibrio sp. at 6 h incubation time, mainly in the group of combination with EC.

Utilizing pomegranate extracts for enhancing yogurt quality and preservation

As consumers become increasingly conscious of synthetic preservatives, the demand for natural antioxidant and antibacterial agents is rising. This study investigates the potential of pomegranate (Punica granatum) extracts to control yogurt spoilage. Methanolic and ethanolic extracts of pomegranate peel, seed, and leaf were analyzed for total phenolic content (TPC), antioxidant activity, and inhibitory effects on ‘yogurt starter strains’ and selected food-related bacteria (Bacillus cereus, Staphylococcus aureus, Staphylococcus sciuri, Escherichia coli, Salmonella enterica subsp. enterica serovar Typhi, and Shigella flexneri). Yogurt samples treated with pomegranate extracts were stored at 4 °C for 28 days. Sensory evaluation was performed on fresh yogurt samples. Physicochemical properties (pH, titratable acidity (TTA), and antioxidant activity) and microbiological changes (aerobic plate count (APC), coliform count, lactic acid bacteria (LAB) count, and total yeast and mold count (TYMC)) were examined weekly. Among the extracts tested, methanolic and ethanolic extracts of peel had the highest TPC (603 and 576.9 mg GAE/g) and the strongest antioxidant activity with the lowest IC50 (141.8 and 183.43 µg/mL). Overall, pomegranate peel, seed, and leaf extracts inhibited foodborne bacteria more effectively than probiotic strains, indicating significant antibacterial potential against spoilage microorganisms. Based on a minimum inhibitory concentration (MIC) test, ethanolic pomegranate extracts were incorporated into lab-made yogurt at 8 mg/mL. Throughout the storage period, all yogurt samples maintained a standard pH (≤ 4.6) and TTA (> 0.5 %). Yogurt fortified with pomegranate peel and leaf extracts exhibited greater antioxidant activity and a significantly lower aerobic plate count while retaining LAB probiotic levels (≥ 106 CFU/g), ensuring safety for consumption (TYMC ≤ 102 CFU/g) and acceptability in terms of five selected quality attributes. Consequently, pomegranate peel and leaf extracts present a promising natural alternative to potassium sorbate in yogurt preservation, maintaining probiotic properties without compromising quality.

Efficacy of grapefruit seed extract and lactic acid bacteria in reducing Clostridioides difficile infection via gut microbiota modulation

Antibiotic-induced dysbiosis is a key factor that causes Clostridioides difficile infection (CDI). The current first-line drugs for initial CDI are antibiotics, and finding an alternate therapy to prevent recurrent CDI is essential. In this study, the anti-C. difficile activities of two isolated probiotic strains (Pediococcus pentosaceus CD5 and Lactobacillus salivarius CD4) and grapefruit seed extract (GSE) were investigated in vitro and in vivo. Specifically, GSE inhibited the growth of C. difficile at a minimal inhibitory concentration (MIC) of 40 ppm. Similarly, live cells of the two probiotics, as well as their cell-free culture supernatants, with or without pH adjustment, substantially suppressed C. difficile growth. In a CDI mouse model, C. difficile colonization in the cecum and colon inflammation were significantly reduced by GSE, followed by the probiotic treatments. A metagenomic analysis of cecal feces revealed that GSE and probiotic treatments modulated gut microbial compositions and enhanced the relative abundances of butyrate-producing bacteria. Therefore, GSE and the novel probiotics can potentially be administered as alternative therapies to alleviate CDI by modifying the gut microbiota.

The Impact of Makgeolli Consumption on Gut Microbiota: An Enterotype-Based Preliminary Study.

Makgeolli, a traditional Korean liquor, contains components such as lactic acid bacteria and dietary fiber, which can induce changes in the gut microbiome. Since variations in microbiome responses may exist between enterotypes-classifications based on the dominant bacterial populations in the gut-we hypothesized that the consumption of makgeolli leads to enterotype-dependent differences in gut microbial structures among healthy participants. This study aimed to determine the effect of makgeolli consumption on gut microbial structures by stratifying all participants into two enterotype groups: Bacteroides-dominant type (B-type, n = 7) and Prevotella-dominant type (P-type, n = 4). The B-type showed an increase in alpha diversity, while no significant difference was observed in the P-type following makgeolli consumption. The composition of gut microbiota significantly changed in the B-type, whereas no noticeable alteration was observed in the P-type after makgeolli consumption. Notably, Prevotella exhibited the most significant changes only in the P-type. In line with the increased abundance of Prevotella, the genes associated with carbohydrate metabolism, including pentose/glucuronate interconversions, fructose/mannose metabolism, starch/sucrose metabolism and amino sugar/nucleotide sugar metabolism were significantly enriched following makgeolli consumption in the P-type. These findings suggest that makgeolli consumption induces enterotype-dependent alterations in gut microbial composition and metabolic pathways, highlighting the potential for personalized dietary interventions.

In vitro Biocontrol of Aflatoxin B1 Through Reduction Using Lactic Acid Bacteria for Animal Heath

In vitro Biocontrol of Aflatoxin B1 Through Reduction Using Lactic Acid Bacteria for Animal Heath

Silage Making of Napier Grass and Sugarcane Top at Different Proportions: Evolution of Natural Fermentation Characteristics, Chemical Composition, and Microbiological Profile

The co-ensiling technique is widely used to improve silage quality; however, it remains unclear as to what high-quality silages can be made by co-ensiling Napier grass (NG) with Sugarcane top (ST). The aim of this study was to evaluate the fermentation characteristics, chemical composition, and microbiological profile of silage produced from mixtures of NG and ST in varying ratios. Silage was prepared using a small-scale fermentation system, and treatments were designed as control silage (NG ensiled alone) or with 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, and 100%ST on a fresh matter basis with six replicates, respectively. Increasing ST in the silage reduced the contents of crude protein, ash, acetic acid, butyric acid, ammonia-N, as well as pH, but increased the contents of dry matter, ether extract, neutral detergent fiber, acid detergent fiber, water-soluble carbohydrate, lactic acid, and lactic acid bacteria. Lactobacillales and Enterobacterales were the dominant orders, with Lactiplantibacillus and Weissella as the dominant genera. Co-ensiling NG with ST enhanced microbial diversity and richness. ST, as a local by-product, is a viable additive to improve NG silage quality and nutrition. This study suggests that good-quality silages can be produced with NG: ST ratios of 40:60 to 20:80 and that these silages offer an opportunity to optimize the nutrient supply for ruminants.

Sumac extract as a natural preservative in mayonnaise: Effects on lipid oxidation, microbial growth, physicochemical, rheological, and sensory characteristics

The present study was conducted to evaluate the effect of sumac extract (SE) as a natural antioxidant and antimicrobial agent on the quality characteristics of low-fat mayonnaise during 63 days of storage. The SE was used at levels of 1 % (SE1%), 3 % (SE3%), and 5 % (SE5%) in the formulations. The two control treatments were produced with chemical preservatives (C1) and without preservatives (C2). The SE caused a significant decrease in peroxide and anisidine values compared to controls after 63 days (4.73–5.77 meq/kg vs. 6.33–6.73 meq/kg for peroxide values, and 6.29–11.21 vs. 12–13.60 for anisidine values, P < 0.05). The higher inhibitory effects of SE on microbial growth in mayonnaises were revealed after 63 days, where the total viable count declined from the highest value (36.5 log CFU/g) in C2 to the lowest count (12 log CFU/g) in SE5%. The yeasts and molds count in SE1% (4.5 log CFU/g) were lower than the counts of 20.5 log CFU/g and 23.5 log CFU/g observed in C1 and C2, respectively. Moreover, significant reductions in Lactic Acid Bacteria were observed in SE-enriched treatments (P < 0.05). The phenolic and flavonoid contents of mayonnaise treatments besides antioxidant capacity were enhanced by increasing SE concentrations. However, the lowest storage modulus and complex viscosity were observed in SE5%. The highest overall acceptance belonged to C1 followed by SE1% at the end of storage. Finally, SE has the potential to be used as a natural preservative in mayonnaise.

Lactic acid bacteria supplementation: A bioprotective approach to mitigating cadmium-induced toxicity and modulating gene expression in murine models

This study aimed to assess the effects of different strains of lactic acid bacteria, namely LeviLactobacillus brevis (AC10), Lacticaseibacillus rhamnosus (AC11), and Pediococcus acidilactici (AC15), on mice exposed to cadmium-induced oxidative stress. The study assessed weight gain, liver enzymes, antioxidant enzymes, immunoglobulin factors, lipid peroxidation, and gene expression in liver and brain of mice. The findings revealed that the AC10 and AC11 strains had a higher ability to absorb Cd as compared to AC15. The in vivo analysis demonstrated that the dietary dual supplementation of AC10 and AC11 resulted in significant (p < 0.05) improvements, including increased body weight and food intake, reduced cadmium tissue deposition, decreased lipid peroxidation, enhanced cellular antioxidant redox potential, suppressed inflammation genes in the liver and brain tissues, and improved morpho-characteristics of the jejunum in mice challenged by cadmium-induced toxicity. The multiple mechanisms of action, including heavy metal sequestration, antioxidant enhancement, and maintenance of intestinal integrity, highlight the potential of these probiotics’ intervention as a viable approach to counteract the deleterious effects of cadmium exposure.

Safety Assessment and Evaluation of Probiotic Potential of Lactobacillus bulgaricus IDCC 3601 for Human Use

Lactic acid bacteria (LAB) are probiotic microorganisms widely used for their health benefits in the food industry. However, recent concerns regarding their safety have highlighted the need for comprehensive safety assessments. In this study, we aimed to evaluate the safety of L. bulgaricus IDCC 3601, isolated from homemade plain yogurt, via genomic, phenotypic, and toxicity-based analyses. L. bulgaricus IDCC 3601 possessed a single circular chromosome of 1,865,001 bp, with a GC content of 49.72%, and 1910 predicted coding sequences. No virulence or antibiotic resistance genes were detected. Although L. bulgaricus IDCC 3601 exhibited antibiotic resistance to gentamicin and kanamycin, this resistance is an intrinsic feature of this species. L. bulgaricus IDCC 3601 did not produce biogenic amines and did not exhibit hemolytic activity. Phenotypic analysis of enzyme activity and carbohydrate fermentation profiles revealed the metabolic features of L. bulgaricus IDCC 3601. Moreover, no deaths or abnormalities were observed in single-dose oral toxicity tests, suggesting that L. bulgaricus IDCC 3601 has no adverse effect on human health. Finally, L. bulgaricus IDCC 3601 inhibited the growth of potential carbapenem-resistant Enterobacteriaceae. Therefore, our results suggest that L. bulgaricus IDCC 3601 is a safe probiotic strain for human consumption.

Innovative Approaches to Camembert Cheese: Optimizing Prebiotics and Coagulation Conditions for Enhanced Quality and Nutrition

The objective of this study is to investigate how different factors, such as lactic acid bacteria, prebiotics (flaxseed powder, watercress seed powder, okra mucilage), and coagulation temperature influence the final quality of curd by conducting three optimization experiments and implementing a structured experimental plan. In the first phase, milk coagulation was assessed at 45 °C with various combinations of lactic acid bacteria (probiotics) and prebiotics (powdered flaxseed and watercress). In the second investigation phase, the effects of lowered probiotic and prebiotic (powdered flaxseed and watercress) concentrations were examined at the coagulation temperature of 38 °C. We investigated the concentration of lactic acid bacteria at 3 mg/mL of milk and the effects of temperature and prebiotics (okra mucilage and flaxseed powder). We observed short milk clotting time (2 s) using the optimized mixture (0.18 mg of probiotics, 1.5 mg of flaxseed powder, and 1.147 mg of watercress powder) per 10 mL of milk. It contrasts with the classical coagulation way optimized at (5.9 and 9.5 s), which were generated at optimal temperatures of 45 and 45.7 °C, respectively. Our new mixture improves the fermentation process of camembert cheese at 38 °C. This cheese had a high flavonoid content, fewer lactic bacteria and molds, a homogeneous texture, and no outer crust, and exceptional sensory attributes such as a creamy and fluid paste. These attributes suggest its potential benefits as a dairy product for individuals with cardiovascular and gastrointestinal conditions.

Functional and physicochemical characterization of a novel pearl millet—Soy milk‐based synbiotic beverage

AbstractThis study presents the physicochemical and functional characterization of a novel fermented pearl millet–soy milk‐based synbiotic beverage. The synbiotic fermented pearl millet beverage was optimized at 40% pearl millet and 10.41% sugar, with hedonic score of more than eight for all the responses, through response surface methodology. The fermented synbiotic beverage, with Lactiplantibacillus plantarum strains exceeding 108 colony‐forming units/mL, exhibited superior nutritional and physicochemical qualities when compared with the unfermented counterpart. The viability percent of lactic acid bacteria in the developed beverage was found to be significantly high (87.83%). The survivability of probiotic culture was 76.71% in simulated gastric juice and 73.87% in simulated intestinal juice after sample storage for 25 days. The result showed that millets stand out as a promising raw material for developing fermented nondairy synbiotic beverages, particularly when combined with microbial strains such as L. plantarum.

The Benefits and Applications of Lactobacillus plantarum in Food and Health: A Narrative Review

Lactobacillus plantarum is a type of gram-positive lactic acid bacteria. This bacterium is considered a safe probiotic and, many applications and benefits including prolonging food shelf-life, enhancing antioxidant activity, improving food flavor characteristics and antimicrobial activities in the food industry, and application as a potential starter for dairy products have been attributed to it. Various studies have also emphasized its health-giving properties. As a result, the features and wide application of this bacterium, as well as the safety of L. plantarum and its strains, have made it a popular probiotic in the food and medical industries. Thus, in the present study keywords including L. plantarum and Lactiplantibacillus plantarum along with application, benefits, food, health, anti-oxidant, anti-diabetic, anti-obesity, anti-inflammatory, antiviral, and anti-depression were searched in databases of PubMed, Scopus, Web of Science, Sience direct and Google Scholar with no time restriction. Then, important features, benefits, and uses of L. plantarum were categorized and discussed. The ability of L. plantarum on the food such as prolonging food shelf-life, enhancing antioxidant activity, improving food flavor characteristics and antimicrobial activities in the food industry, and as a potential starter for dairy products is effective. In addition, several studies have emphasized of L. plantarum health-giving properties

Comprehensive evaluation of Flammulina velutipes residues polysaccharide based on in vitro digestion and human fecal fermentation

Flammulina velutipes residues (FVR) are the waste culture medium derived from the collection of Flammulina velutipes fruiting bodies, with an annual output that remains largely unexplored. The characteristics of digestion and fermentation of Flammulina velutipes residues polysaccharide (FVRP) are still relatively unknown. This study investigated the structure of the gut microbiota through 16 s rDNA gene sequencing and analyzed changes in short-chain fatty acid (SCFA) content via targeted metabolome analysis. The aim was to explore the prebiotic activity of FVRP based on a simulated digestion model combined with an in vitro anaerobic fermentation model. The results demonstrated that FVRP did not exhibit significant changes during in vitro digestion and fermentation but did enhance antioxidant activity. Furthermore, FVRP was found to rapidly reduce the pH value and increase SCFA production in the fermentation broth from lactic acid bacteria and human feces. Notably, FVRP altered the gut microbiota structure, significantly increasing the relative abundance of Firmicutes and Bacteroidota. Thus, FVRP could be considered a promising prebiotic food and feed additive that promotes the generation of short-chain fatty acids by modulating gut microbiota.

Viability of Lactobacillus acidophilus in acidophilus milk during frozen storage and its potential to lower cholesterol: an In vivo study.

The viability and functional activity of lactic acid bacteria (LAB), such as Lactobacillus acidophilus in a fermented milk product is important. One of the functional activities of the LAB in fermented milk is the ability of the LAB to positively impact the human health. This study aimed to determine the effect of frozen storage on the fermented milk, namely acidophilus milk, based on the nutritional milk quality, the viability of L. acidophilus, and its potential to lower cholesterol levels in Wistar rats. The parameters measured was including milk quality (mainly pH, protein content, lactic acid levels, and syneresis), L. acidophilus bacterial viability in frozen storage (for 1, 2, and 3months), and some biological assays to evaluate the potential of milk in lowering blood cholesterol levels in Wistar rats. The result of this study suggests that acidophilus milk quality can be maintained in frozen storage for two months, and it had lactic acid levels of 1.07%, pH of 4.08, and protein levels of 3.33%. Giving acidophilus milk to Wistar rats for 15days could reduce the cholesterol level continuously until 30days of treatment. Therefore, this study proves that acidophilus milk quality can be maintained very well in frozen storage, and its functional properties to lower the cholesterol level of Wistar rats can be achieved after two weeks of consumption.

Lactic Acid Bacteria Viability and Sensory Evaluation of Yogurt Prepared From Different Breeds Goat Milk

Lactic Acid Bacteria Viability and Sensory Evaluation of Yogurt Prepared From Different Breeds Goat Milk

Bacillus clausii: A Review into Story of Its Probiotic Success and Potential Food Applications

Probiotics are live microbiota that can confer their hereditary health benefits upon the host. They can positively alter the diversity of the host’s gut microbiota population. Bacillus clausii is a spore-producing potential probiotic. Its application in the food industry has been highly recommended. Clausii spores are greatly resistant to harsh food processing treatment (heat and industrial pressure) and they can maintain their physiological traits (acid, bile salt) inside the human gut. The utilization of spores can enhance the nutritional viability of livestock, as well as the functionality of on-shelf products. This contemporary review covers the great attributes of B. clausii, including physiological characteristics, modes of action, probiotics benefits, a safety assessment, and the bacteria food industry applications for the purposes of producing enhanced functional foods known as probiotic foods. To our knowledge, although B. clausii has been widely applied in food industry, the amount of the literature that is dedicated to its role in sustainable food production and addresses its potential sporogenous probiotic behavior is fewer as compared to non-sporogenous lactic acid bacteria (Lactobacillus spp. and Bifidobacterium spp.). In our review, we also discovered a lack of empirical evidence on the synbiotic and synergistic behavior of clausii in combination with other active nutrients. Therefore, our review suggests that more studies should be conducted on the potential probiotic contribution of B. clausii in sustainable food production and its synergistic and synbiotic behavior in conjunction with other active nutrients.

Ameliorated Viability of Lactic Acid Bacteria in Fruit Juice Isolated from Indigenous Dahi with Prebiotics (Asparagus falcatus and Zingiber officinale)

Dahi is a fermented milk product containing probiotic lactic acid bacteria. This study aimed to isolate, identify, and characterize lactic acid-producing bacteria from native Dahi and evaluate their viability in orange juice using natural prebiotics. Dahi samples were obtained from local shops in Chattogram and Bogura, Bangladesh. Lactic acid-producing bacteria were isolated using MRS (de Mann Rogosa and Sharpe) medium. The isolated bacteria were identified through colony morphology, biochemical tests, and probiotic characteristics. Molecular identification was performed using polymerase chain reaction (PCR) targeting conserved 16S rDNA regions. Isolates of the genus Lactobacillus and Lactococcus lactis sp. Lactis were confirmed and used to develop probiotic orange juice. Prebiotics (Asparagus falcatus and Zingiber officinale) were added to the juice to support probiotic growth. The inoculated cell’s viability and the juice’s physicochemical parameters were evaluated during fermentation (48 hours) and storage (28 days). All fruit juice samples showed a mean number of viable cells of at least 1×105 CFU/mL during the 48-hour fermentation and 28-day storage in the refrigerator. Using natural prebiotics positively affected the survival of lactic acid bacteria, as demonstrated by bacterial colony growth on Petri dishes. Developing probiotic fruit juice enriched with prebiotics could be an effective alternative for individuals allergic or intolerant to milk-based products. Incorporating lactic acid bacteria from native Dahi into orange juice, combined with natural prebiotics, resulted in viable probiotic cells throughout fermentation and storage.

Antioxidant Activity and Other Characteristics of Lactic Acid Bacteria Isolated from Korean Traditional Sweet Potato Stalk Kimchi.

The aim of this study was to examine the biological activity and probiotic properties of lactic acid bacteria (LAB) isolated from sweet potato stalk kimchi (SPK). Various LAB and Bacillus spp. are active in the early stages of the fermentation of kimchi made from sweet potato stalk. Four strains of LAB were identified, including SPK2 (Levilactobacillus brevis ATCC 14869), SPK3 (Latilactobacillus sakei NBRC 15893), SPK8 and SPK9 (Leuconostoc mesenteroides subsp. dextranicum NCFB 529). SPK2, SPK3, SPK8, and SPK9 showed 64.64-94.23% bile acid resistance and 78.66-82.61% pH resistance. We identified over 106 CFU/mL after heat treatment at 75 °C. Four strains showed high antimicrobial activity to Escherichia coli and Salmonella Typhimurium with a clear zone of >11 mm. SPK2 had the highest antioxidative potentials, higher than the other three bacteria, with 44.96 μg of gallic acid equivalent/mg and 63.57% DPPH scavenging activity. These results demonstrate that the four strains isolated from sweet potato kimchi stalk show potential as probiotics with excellent antibacterial effects and may be useful in developing health-promoting products.

Effect of Lentilactobacillus buchneri on Chemical and Microbial Compositions of Herba Leonuri (Leonurus japonicus Houtt.)-Contained Alfalfa Silage

Lactic acid bacteria (LAB) inoculants are commonly used in silage production, yet their effects on silage containing antimicrobial components, such as those found in Leonurus japonicus, remain less explored. Herein, the harvested alfalfa were thoroughly mixed with dried Leonurus japonicus Houtt. (LJH) at a ratio of 9:1 on a fresh weight basis and treated without (CK) or with a lactic acid bacterial inoculant (L; Lentilactobacillus buchneri). The mixtures were stored under anaerobic conditions in vacuum-sealed polyethylene bags for 30 days at ambient temperature. The L-treated silage exhibited high levels of water-soluble carbohydrates (4.98% dry matter (DM)) and acid detergent fiber (27.88% DM). Compared to that of treatment CK, treatment with L increased the acetic acid content of the silage, as a result of increased (p &lt; 0.05) bacterial dominance and decreased (p &lt; 0.05) bacterial richness indices (e.g., Pielou’s E, Shannon, and Simpson) in the pre-storage period. However, these changes gradually reduced as the storage length increased. Treatment L reshaped the bacterial community structure of silage, by increasing the prevalence of Lactobacillus and reducing relative abundances of Enterococcus and Weissella. However, the principal coordinate and Bray–Curtis index analyses illustrated that samples from the L-treated silages exhibited similarities to the CK samples post-fermentation. Overall, the effect of LJH on LAB was only observed in the later stages of fermentation, which did not sufficiently change the silage quality. Hence, using LJH in silage is vital for clean livestock production without compromising the function of LAB when mixed with alfalfa silage.