Biomedicine & Pharmacotherapy

Systematic Review: Preservative and Therapeutic Efficacy of Probiotic Lactic Acid

Bacteria
--Manuscript Draft--

Manuscript Number:

Article Type: Review Article

Keywords: Probiotic Lactic Acid Bacteria; Preservative Efficiency; Therapeutic Efficacy; Food
Preservation; and Gut Health

Corresponding Author: Desalegn Amenu

Wollega University Department of Biology
ETHIOPIA

First Author: Desalegn Amenu

Order of Authors: Desalegn Amenu

Ayantu Nugusa

Temesgen Tafesse

Abstract: Introduction: Probiotic lactic acid bacteria (LAB) play pivotal roles in both food
preservation and human health, leveraging their antimicrobial properties and
therapeutic benefits.
Methodology: A thorough search was conducted across PubMed, Google Scholar, and
ScienceDirect databases, covering studies published from 2010 to 2023.
Results and discussion: In parallel, LAB exhibit substantial therapeutic potential by
modulating gut microbiota, enhancing intestinal barrier function, and boosting immune
responses. Clinical trials and in vivo studies highlight their effectiveness in preventing
gastrointestinal infections, reducing serum cholesterol levels, and providing antioxidant
protection. This review underscores the need for further research to optimize LAB
strains for specific preservative and therapeutic applications. Overall, LAB represent a
valuable asset in both the food industry and medical field, offering natural solutions for
food preservation and health improvement. Future studies should focus on elucidating
the precise mechanisms of action and developing targeted probiotic formulations to
maximize their benefits.
Challenges and Future Directions: Variability in LAB efficacy based on strain
specificity, dosage, and administration mode.
Conclusion: Probiotic LAB represent a dual-purpose solution in the food industry and
healthcare, offering natural alternatives for food preservation and health improvement.
Future studies should focus on refining probiotic formulations, understanding their
mechanisms of action, and optimizing their applications in both food technology and
clinical practice. This review underscores the promising potential of LAB as versatile
agents in promoting both food safety and human well-being.

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Cover Letter

Key Points:

1. Mechanisms of Preservation: Lactic acid bacteria (LAB) preserve fruits and vegetables through
acid production, bacteriocin secretion, competition for nutrients, and exopolysaccharide
production. These mechanisms inhibit spoilage organisms and pathogenic bacteria.

2. Extended Shelf Life: LAB significantly extend the shelf life of produce by lowering pH levels and
creating an environment hostile to spoilage organisms and pathogens. This leads to longer-
lasting fruits and vegetables.

3. Nutritional and Sensory Enhancement: The fermentation process mediated by LAB not only
preserves but also enhances the nutritional value and sensory properties (taste, texture) of
fruits and vegetables, contributing to healthier and more appealing food products.

4. Biotechnological Advances: Advances in biotechnology have resulted in the development of

optimized LAB strains and starter cultures. These advancements enhance the efficacy of
preservation processes, making them more efficient and effective.

5. Applications and Examples: Notable applications of LAB in food preservation include the
fermentation of cabbage into sauerkraut, the production of kimchi, and the pickling of
cucumbers. These examples demonstrate the practical benefits and versatility of LAB in
preserving various types of produce.
Review Proposal Form

Systematic Review Proposal Form

Title: Preservative and Therapeutic Efficacy of Probiotic Lactic Acid Bacteria

1. Introduction: Probiotic lactic acid bacteria (LAB) are widely recognized for their beneficial
effects on human health and food preservation. This systematic review aims to evaluate the
preservative properties and therapeutic efficacy of probiotic LAB in various applications,
including food products and clinical settings. The review will synthesize current evidence to
provide a comprehensive understanding of the role of probiotic LAB.

2. Objectives:

 To assess the preservative efficacy of probiotic LAB in food products.

 To evaluate the therapeutic benefits of probiotic LAB in clinical settings.
 To identify the mechanisms through which probiotic LAB exert their preservative and
therapeutic effects.

3. Research Questions:

 What is the preservative efficacy of probiotic LAB in different types of food products?
 What therapeutic benefits do probiotic LAB provide in clinical applications?
 What mechanisms are involved in the preservative and therapeutic actions of probiotic
LAB?

4. Methodology:

4.1. Eligibility Criteria:

 Types of Studies: Randomized controlled trials (RCTs), cohort studies, case-control

studies, and observational studies.
 Participants: Studies involving human subjects for therapeutic efficacy and food
products for preservative efficacy.
 Interventions: Use of probiotic LAB in food preservation or as a therapeutic agent.
 Outcomes: Measures of preservative efficacy (e.g., shelf life, microbial load) and
therapeutic outcomes (e.g., gastrointestinal health, immune response).

4.2. Information Sources:

 Databases: PubMed, Cochrane Library, Scopus, Web of Science, and Google Scholar.
 Other Sources: Reference lists of selected studies, grey literature, and conference
proceedings.

4.3. Search Strategy: A comprehensive search strategy will be developed using keywords such
as "probiotic lactic acid bacteria," "preservative efficacy," "therapeutic efficacy," "food
preservation," and "clinical benefits." Boolean operators AND, OR, and NOT will be used to
combine search terms. The search will be limited to studies published in English from 2000 to
the present.

4.4. Data Extraction: Data will be extracted using a standardized form, including study
characteristics (e.g., author, year, study design), participant characteristics, intervention details,
and outcomes. Two reviewers will independently extract data to ensure accuracy and
consistency.

4.5. Risk of Bias Assessment: The Cochrane Risk of Bias tool will be used to assess the quality
of included studies. Studies will be evaluated for selection bias, performance bias, detection bias,
attrition bias, and reporting bias.

4.6. Data Synthesis: Data will be synthesized qualitatively and, where possible, quantitatively
using meta-analysis. Subgroup analyses will be conducted based on the type of food product,
clinical condition, and LAB strain.

5. Timeline:

 Protocol Development: 1 month

 Literature Search: 2 months
 Screening and Selection: 2 months
 Data Extraction and Analysis: 3 months
 Writing and Review: 2 months
 Submission: 1 month

6. Dissemination Plan: The findings of this systematic review will be disseminated through
peer-reviewed journal publications, conference presentations, and stakeholder meetings.

7. Ethical Considerations: This systematic review will use data from previously published
studies, so no ethical approval is required. However, ethical considerations related to data
handling and reporting will be strictly followed.

8. Review Team:

 Lead Investigator: [Name, Affiliation]

 Co-Investigators: [Names, Affiliations]
 Methodologist: [Name, Affiliation]
 Statistical Analyst: [Name, Affiliation]
 Research Assistants: [Names, Affiliations]

9. Funding: [Details of funding sources or a statement indicating no specific funding was

received for this review.]

10. Conflicts of Interest: The review team will declare any potential conflicts of interest.

11. References: A preliminary list of key references will be included in the proposal.
Manuscript File Click here to view linked References

1 Systematic Review: Preservative and Therapeutic Efficacy of Probiotic Lactic

2 Acid Bacteria
3 Desalegn Amenu1, Ayantu Nugusa1 and Temesgen Tafesse2
4 1. Wollega University, College of Natural and Computational sciences, Biology
5 Department, Nekemte, Ethiopia.
6 2. Armuear Hansen Research Institute, Microbiology and Microbial-biotechnology, Addis
7 Ababa, Ethiopia.

8 Abstract

9 Title: Systematic Review: Probiotic Lactic Acid Bacteria (LAB) in Food Preservation and
10 Therapeutic Applications

11 Introduction: Probiotic lactic acid bacteria (LAB) play pivotal roles in both food preservation and
12 human health, leveraging their antimicrobial properties and therapeutic benefits. This systematic
13 review synthesizes recent research to comprehensively assess the preservative efficiency and
14 therapeutic efficacy of LAB.

15 Methodology: A thorough search was conducted across PubMed, Google Scholar, and
16 ScienceDirect databases, covering studies published from 2010 to 2023. Selection criteria included
17 relevance, methodological rigor, and quality of evidence.

18 Results and discussion: LAB demonstrate significant efficacy in extending the shelf life and
19 ensuring the safety of various food products, such as dairy, fermented vegetables, and meat
20 products. In parallel, LAB exhibit substantial therapeutic potential by modulating gut microbiota,
21 enhancing intestinal barrier function, and boosting immune responses. Clinical trials and in vivo
22 studies highlight their effectiveness in preventing gastrointestinal infections, reducing serum
23 cholesterol levels, and providing antioxidant protection. Despite the promising evidence, the
24 efficacy of LAB varies significantly depending on the strain, dosage, and mode of administration.
25 This review underscores the need for further research to optimize LAB strains for specific
26 preservative and therapeutic applications. Overall, LAB represent a valuable asset in both the
27 food industry and medical field, offering natural solutions for food preservation and health
28 improvement. Future studies should focus on elucidating the precise mechanisms of action and
29 developing targeted probiotic formulations to maximize their benefits.
30 Challenges and Future Directions: Variability in LAB efficacy based on strain specificity,
31 dosage, and administration mode. Need for further research to optimize LAB strains for targeted
32 food preservation and therapeutic applications. Elucidating precise molecular mechanisms
33 underlying LAB actions in both food and health contexts.

34 Conclusion: Probiotic LAB represent a dual-purpose solution in the food industry and healthcare,
35 offering natural alternatives for food preservation and health improvement. Future studies should
36 focus on refining probiotic formulations, understanding their mechanisms of action, and
37 optimizing their applications in both food technology and clinical practice. This review
38 underscores the promising potential of LAB as versatile agents in promoting both food safety and
39 human well-being.

40 Key words: Probiotic Lactic Acid Bacteria, Preservative Efficiency, Therapeutic Efficacy, Food
41 Preservation, and Gut Health.

42 1. Introduction

43 Probiotic lactic acid bacteria (LAB) have garnered significant attention due to their dual roles in
44 food preservation and therapeutic applications. These microorganisms, commonly found in
45 fermented foods, possess the ability to inhibit the growth of pathogenic bacteria through the
46 production of organic acids, bacteriocins, and other antimicrobial substances(Khushboo et al.,
47 2023; Zapaśnik et al., 2022). Their preservative efficacy not only extends the shelf life of food
48 products but also enhances their safety and nutritional value. In addition to their preservative
49 properties, LAB exhibit numerous health benefits, including the modulation of gut microbiota,
50 enhancement of immune response, and potential therapeutic effects against gastrointestinal
51 disorders, infections, and even certain chronic diseases (Cotter et al., 2005; Umu et al., 2017).

52 The preservative efficacy of LAB is primarily attributed to their metabolic activities, which lead
53 to the production of lactic acid and other organic acids(Zapaśnik et al., 2022). These acids lower
54 the pH of the food environment, creating conditions unfavorable for spoilage and pathogenic
55 microorganisms(Cotter et al., 2005). Furthermore, LAB can produce bacteriocins—proteinaceous
56 substances with bactericidal properties that target specific bacterial strains. For instance, nisin, a
57 well-known bacteriocin produced by Lactococcus lactis, is widely used in the food industry for its
58 effectiveness against a broad spectrum of Gram-positive bacteria(Anumudu et al., 2021). These
59 dual mechanisms highlight the pivotal role of LAB in enhancing the shelf life and safety of food
60 products, making them indispensable in food preservation practices. Continued research into the
61 metabolic pathways and applications of LAB promises further innovations in food technology and
62 microbiology, ensuring both quality and safety in the global food supply chain.

63 The therapeutic potential of LAB is equally compelling (Rezvani et al., 2016). They contribute to
64 the maintenance of a healthy gut microbiota, which is essential for proper digestion, nutrient
65 absorption, and immune function. Studies have shown that specific strains of LAB can alleviate
66 symptoms of irritable bowel syndrome (IBS), reduce the incidence of antibiotic-associated
67 diarrhea, and improve outcomes in inflammatory bowel disease (IBD) (Distrutti et al., 2016;
68 Skoufou et al., 2024).Moreover, LAB have been investigated for their potential to prevent and
69 manage infections, particularly those involving the gastrointestinal and respiratory tracts(Petrariu
70 et al., 2023). Emerging evidence also suggests a role for LAB in modulating the immune system
71 and reducing the risk of certain chronic diseases, such as cardiovascular disease and diabetes (Zhao
72 et al., 2023). Hence, the diverse therapeutic applications of LAB underscore their significant role
73 in enhancing human health beyond food preservation. Continued research into their mechanisms
74 and clinical efficacy promises to further illuminate their potential in preventive and therapeutic
75 medicine.

76 In summary, the preservation effectiveness of LAB hinges on their ability to produce lactic acid
77 and other organic acids, which create an acidic environment detrimental to spoilage and pathogenic
78 microorganisms (Zapaśnik et al., 2022; Cotter et al., 2005). Additionally, LAB contribute to food
79 safety by synthesizing bacteriocins such as nisin, which exhibit targeted antibacterial activity
80 against specific types of bacteria, notably Gram-positive species (Anumudu et al., 2021).

81 Understanding these mechanisms is crucial for several reasons. First, it underpins the scientific
82 basis for using LAB in food preservation, extending shelf life and ensuring product safety without
83 relying on synthetic additives. Second, the ability of LAB to produce natural antimicrobial
84 compounds like bacteriocins offers a sustainable alternative to traditional preservatives, aligning
85 with consumer preferences for natural and minimally processed foods. Finally, ongoing research
86 into LAB metabolism and bacteriocin production holds promise for developing novel food
 87 preservation strategies that are effective, eco-friendly, and compatible with modern food
88 production practices. In essence, the study of LAB's preservative abilities not only enhances our
89 understanding of microbial ecology in food systems but also contributes to advancing food
90 technology towards safer and more sustainable practices.

91

92

93

94

95

96

97 2. Methodology
98 Search Strategy and Databases Utilized:
99  Databases:
100 o PubMed, Scopus, Web of Science, Cochrane Library, Embase.
101 o These databases were chosen for their comprehensive coverage of biomedical
102 literature, systematic reviews
103  Search Terms:
104 o Keywords related to "probiotic lactic acid bacteria", "preservation", "therapeutic
105 efficacy"
106 o Boolean operators (AND, OR) was used to combine these keywords effectively to
107 capture relevant studies.
108  Date and Language Restrictions:
109 o Articles published from 2010 to 2024 were included to focus on recent
110 advancements in probiotic LAB research.
111 o Only articles published in the English language was included due to resource
112 constraints.
113 Screening and Selection Process of Articles:
114  Initial Screening:
115 o Titles and abstracts of identified articles were screened independently by two
116 reviewers.
117 o Articles deemed potentially relevant based on inclusion criteria were proceed to
118 full-text review.
119  Full-Text Review:
120 o Full-text articles were retrieved and reviewed to assess eligibility for final inclusion
121 in the systematic review.
122 o Reasons for exclusion of articles were documented, such as irrelevant outcomes or
123 non-human studies.
124
125 3. Preservative efficiency of probiotics

126 The preservative efficiency of probiotics, particularly lactic acid bacteria (LAB), is a crucial aspect
127 in food science and industry. Probiotics exert their preservative effects through several key
128 mechanisms:

129 3.1.Acid Production:

130 Probiotic LAB, such as various strains of Lactobacillus and Bifidobacterium, utilize their ability
131 to ferment carbohydrates present in food substrates. Through fermentation, LAB metabolize sugars
132 into lactic acid and other organic acids (Cotter et al., 2005).This metabolic process leads to a
133 decrease in the pH of the food matrix. The lowered pH creates an acidic environment that is
134 inhospitable to the growth and survival of spoilage microorganisms and pathogens (Zapaśnik et
135 al., 2022). The acidity not only directly inhibits microbial growth but also enhances the stability
136 of proteins and other macromolecules in the food, thereby extending its shelf life(Khushboo et al.,
137 2023).

138 3.2.Production of Antimicrobial Peptides:

139 In addition to acid production, probiotic LAB are capable of synthesizing antimicrobial peptides
140 known as bacteriocins. One of the well-known examples is nisin, produced by Lactococcus lactis.
141 Bacteriocins exhibit selective antimicrobial activity against specific bacterial strains, particularly
142 Gram-positive bacteria (Anumudu et al., 2021).These peptides function by disrupting the integrity
143 of bacterial cell membranes, leading to cell lysis and eventual death of the targeted
144 microorganisms(Cotter et al., 2005). This dual mechanism of acidification and bacteriocin
145 production allows probiotic LAB to effectively control microbial populations in food, contributing
146 significantly to its preservation.

147 3.3.Competitive Exclusion:

148 Furthermore, probiotic LAB can outcompete and inhibit the growth of pathogenic or spoilage
149 microorganisms through competitive exclusion. By colonizing food matrices and adhering to
150 surfaces, LAB create a competitive advantage for themselves, limiting the available nutrients and
151 space for other bacteria to thrive (Cotter et al., 2005). This competitive exclusion mechanism
152 further contributes to the preservation of food products by preventing contamination and spoilage.3

153 3.4.Interaction with Food Matrix:

154 The effectiveness of probiotic lactic acid bacteria (LAB) in food preservation depends significantly
155 on their interaction with the specific characteristics of the food matrix. Recent studies underscore
156 several key factors within the food environment that influence LAB's growth, metabolic activity,
157 and ultimately their preservation capabilities (Zapaśnik et al., 2022). LAB are particularly sensitive
158 to pH levels, thriving in mildly acidic conditions typically ranging from pH 4 to 5. This acidity is
159 primarily generated through the fermentation process, where LAB produce lactic acid. Optimal
160 pH conditions are crucial as they support the growth and activity of LAB, enhancing their ability
161 to outcompete spoilage microorganisms and pathogens that prefer neutral or alkaline environments
162 (Cotter et al., 2005).

163 Another critical factor is the moisture content of the food matrix. LAB require sufficient moisture
164 to sustain their metabolic processes and enzymatic activities. In environments with low moisture
165 content, such as dried meats or cereals, LAB may enter a dormant state or exhibit reduced
166 metabolic activity, which limits their effectiveness in preserving the product (Zapaśnik et al.,
167 2022). Nutrient availability also plays a pivotal role in shaping LAB's preservation efficacy.
168 Adequate nutrients, including carbohydrates and nitrogen sources, support robust LAB
169 metabolism. This metabolic activity enables LAB to produce antimicrobial compounds like
170 organic acids and bacteriocins, which inhibit the growth of spoilage bacteria. In nutrient-rich
171 environments, LAB can more effectively contribute to food preservation efforts by controlling
172 microbial populations (Cotter et al., 2005). Additionally, the composition and structure of the food
173 matrix, such as its texture, fat content, and the presence of preservatives, influence LAB activity.
174 For instance, the distribution and survival of LAB within the food product can be influenced by
175 the presence of fats or oils, impacting their ability to uniformly exert preservative effects
176 throughout the matrix (Zapaśnik et al., 2022).

177

178
179 3.5.Antimicrobial activity

180 Antimicrobial activity is a crucial aspect of probiotic lactic acid bacteria's (LAB) role in food
181 preservation. LAB are known to produce antimicrobial compounds, notably bacteriocins, which
182 exhibit targeted bactericidal effects against a wide range of bacteria, particularly Gram-positive
183 strains. These bacteriocins play a significant role in inhibiting the growth of pathogens and
184 spoilage microorganisms, thereby enhancing food safety and extending shelf life (Anumudu et al.,
185 2021).

186 Bacteriocins are proteinaceous peptides synthesized by LAB as a natural defense mechanism
187 against competing microorganisms in their environment. They function by disrupting the cell
188 membranes of target bacteria, leading to cell lysis and subsequent death of the microbial cells
189 (Cotter et al., 2005). This selective antimicrobial action makes bacteriocins effective natural
190 preservatives in food systems, as they specifically target harmful bacteria while sparing beneficial
191 microflora. Studies have demonstrated the efficacy of bacteriocins, such as nisin produced by
192 Lactococcus lactis, against various foodborne pathogens including Listeria monocytogenes and
193 Staphylococcus aureus (Anumudu et al., 2021). The application of bacteriocins in food products
194 helps mitigate the risk of bacterial contamination and spoilage, contributing to improved food
195 quality and safety.

196 Experimental Studies:

197 Experimental models and studies in food systems consistently demonstrate the effectiveness of
198 probiotic LAB in extending the shelf life of various products. For instance, research has shown
199 that supplementation with LAB in fermented dairy products significantly reduces the growth of
200 spoilage bacteria. This is achieved through mechanisms such as pH reduction and production of
201 antimicrobial compounds like bacteriocins, thereby enhancing product stability and safety
202 (Zapaśnik et al., 2022).

203

204
205 Clinical Trials and Field Applications:

206 Clinical trials and field applications provide further evidence of probiotic LAB's preservation
207 efficacy in real-world settings. These studies evaluate LAB's impact on microbial populations,
208 sensory attributes, and consumer acceptability in food products. For example, clinical trials have
209 validated the ability of LAB to maintain food safety by controlling pathogenic bacteria while
210 preserving sensory qualities that are crucial for consumer satisfaction (Cotter et al., 2005).

211 Overall, the combination of experimental evidence and clinical data supports the practical
212 application of probiotic LAB in enhancing food preservation strategies. These findings not only
213 underscore LAB's role in extending shelf life and improving food safety but also highlight their
214 potential as natural alternatives to traditional preservatives in meeting consumer preferences for
215 healthier and minimally processed foods. Continued research aims to further optimize LAB-based
216 preservation methods and expand their application across a broader range of food products.

217 4. Therapeutic Efficacy of LAB

218 The therapeutic efficacy of probiotic lactic acid bacteria (LAB) spans a wide range of health
219 benefits, supported by evidence from various studies and clinical trials. Here's an outline detailing
220 specific aspects and clinical evidence of their therapeutic applications:

221 4.1.Promotion of Beneficial Bacteria by Lactic Acid Bacteria

222 Lactic Acid Bacteria (LAB), including Lactobacillus and Bifidobacterium species, are
223 instrumental in fostering a healthy gut microbiota through diverse mechanisms supported by recent
224 research. Firstly, LAB are adept at adhering to and colonizing the mucosal surfaces of the
225 gastrointestinal tract, facilitating prolonged interactions with the gut microbiota and enhancing
226 their beneficial effects (Pessione, 2012). Secondly, LAB are prolific producers of metabolites that
227 create an optimal environment for beneficial bacteria: Short-chain fatty acids (SCFAs) such as
228 acetate, propionate, and butyrate are key metabolites generated through the fermentation of dietary
229 fibers. These SCFAs not only serve as vital energy sources for intestinal epithelial cells but also
230 contribute significantly to maintaining gut barrier integrity and immune function (Fusco et al.,
231 2023; Silva et al., 2020). Antimicrobial peptides (AMPs) produced by LAB exhibit potent
232 antimicrobial properties. These peptides selectively inhibit the growth of pathogenic bacteria while
233 promoting the growth and proliferation of beneficial bacteria like Bifidobacteria, thereby
234 contributing to gut health (Castillo et al., 2012). Furthermore, LAB play a crucial role in
235 modulating the pH levels of the intestinal environment through the production of lactic acid. This
236 acidification creates an inhospitable environment for many pathogenic bacteria while supporting
237 the growth of acid-tolerant beneficial bacteria(Pessione, 2012)

238 LAB also exert influence over nutrient availability within the gut, enhancing the accessibility of
239 essential compounds that support the growth and vitality of beneficial microbes(Daria & Nigam,
240 2022). Lastly, LAB engage in beneficial cross-feeding interactions with other members of the gut
241 microbiota. For instance, they enzymatically degrade complex carbohydrates into simpler sugars
242 and SCFAs. These breakdown products serve as substrates for neighboring beneficial bacteria that
243 may lack the necessary enzymes for initial degradation, fostering a symbiotic relationship within
244 the gut ecosystem(Flint et al., 2012).

245 4.2.Modulation of Immune Response by Probiotic Lactic Acid Bacteria

246 Probiotic Lactic Acid Bacteria (LAB), including species like Lactobacillus and Bifidobacterium,
247 play a crucial role in modulating immune responses by regulating immune system activity and
248 mitigating inflammation and allergic responses through various mechanisms. Firstly, LAB
249 enhance barrier function within the gastrointestinal tract by fortifying the intestinal epithelial
250 barrier. This reinforcement helps prevent the translocation of pathogens and antigens from the gut
251 into the bloodstream, thereby reducing the overall burden on the immune system and promoting
252 gut homeostasis(Zhang et al., 2016).

253 Secondly, LAB interact with intestinal epithelial cells and immune cells through toll-like receptors
254 (TLRs). This interaction triggers regulatory pathways that balance immune responses,
255 distinguishing between harmful pathogens and beneficial commensal microorganisms. By
256 modulating immune activation, LAB contribute to maintaining immune homeostasis in the
257 gut(Frosali et al., 2015). LAB also exert anti-inflammatory effects by stimulating the production
258 of anti-inflammatory cytokines such as interleukin-10 (IL-10) and transforming growth factor-beta
259 (TGF-β) by immune cells. These cytokines help to suppress excessive immune responses and
260 inflammation within the gut mucosa, contributing to overall gut health (Iyer & Cheng, 2012).
261 Furthermore, LAB can inhibit the nuclear factor-kappa B (NF-kB) pathway in immune cells. This
262 pathway is pivotal in regulating the expression of genes involved in inflammation. By dampening
263 NF-kB activation, LAB mitigate inflammation within the gut, thus supporting immune balance
264 and gut health(Bąska & Norbury, 2022).

265 In terms of allergic responses, LAB influence the balance between T helper 17 (Th17) cells and
266 regulatory T cells (Tregs) in the gut. This balance is critical for immune tolerance and the
267 prevention of allergic reactions. LAB promote the differentiation of Tregs while suppressing the
268 differentiation of Th17 cells, thereby reducing allergic inflammation(Cheng et al., 2019).
269 Moreover, LAB can modulate the production of immunoglobulin E (IgE), an antibody involved in
270 allergic reactions. By decreasing IgE levels, LAB contribute to mitigating allergic sensitization
271 and responses, further highlighting their role in immune modulation(Shamji et al., 2021). In
272 conclusion, probiotic Lactic Acid Bacteria exhibit diverse mechanisms through which they
273 modulate immune responses, reduce inflammation, and alleviate allergic reactions in the gut.
274 These insights underscore their potential therapeutic applications in immune-related disorders and
275 underscore the importance of further research into their mechanisms and clinical efficacy.

276 4.3.Clinical Applications of Probiotic Lactic Acid Bacteria (LAB)

277 4.3.1. Gastrointestinal Health

278 Probiotic Lactic Acid Bacteria (LAB), including species like Lactobacillus and Bifidobacterium,
279 exhibit a wide range of clinical applications across various health domains, particularly in
280 gastrointestinal health. In the realm of gastrointestinal health, LAB have proven effective in
281 managing different types of diarrhea, including antibiotic-associated diarrhea and infectious
282 diarrhea(Petrariu et al., 2023).They play a crucial role in restoring the balance of gut microbiota
283 that is often disrupted by antibiotic use. Additionally, LAB inhibit the growth of pathogenic
284 bacteria, thereby promoting a healthier gut environment (Kesavelu & Jog, 2023).

285 Moreover, probiotic LAB supplementation has shown promise in the management of symptoms
286 associated with Irritable Bowel Syndrome (IBS). Individuals with IBS often experience discomfort
287 such as bloating, abdominal pain, and irregular bowel movements. LAB contribute to improving
288 gut health by reducing inflammation and enhancing bowel function, which can alleviate these
289 distressing symptoms(Didari et al., 2015; More et al., 2020). In conclusion, the therapeutic benefits
290 of probiotic Lactic Acid Bacteria extend to gastrointestinal health through their ability to restore
291 microbiota balance, inhibit pathogenic bacteria, and alleviate symptoms of conditions like diarrhea
292 and IBS. These findings underscore their potential as valuable interventions in clinical practice,
293 supporting overall gut health and well-being.

294 4.3.2. Immune System Support

295 Probiotic Lactic Acid Bacteria (LAB), such as Lactobacillus and Bifidobacterium species,
296 contribute significantly to immune system support through their beneficial effects on respiratory
297 infections, allergies, and autoimmune diseases. LAB enhance mucosal immunity and support
298 respiratory tract health, which helps in both preventing and reducing the severity of respiratory
299 infections like the common cold and influenza (Ahanchian & Jafari, 2016; Mettelman et al., 2022).
300 By bolstering mucosal barriers and promoting immune responses in the respiratory tract, LAB
301 reduce the likelihood of infection and aid in quicker recovery from respiratory illnesses.

302 In terms of allergy management and modulation of autoimmune diseases, LAB play a pivotal role
303 in regulating immune responses. They promote immune tolerance and mitigate allergic reactions
304 by influencing the differentiation of immune cells and modulating cytokine production crucial for
305 maintaining immune balance (Goswami et al., 2022; Mukuku et al., 2019). This modulation helps
306 in reducing allergic sensitization and symptoms, as well as potentially alleviating autoimmune
307 responses by promoting tolerance towards self-antigens.

308 In summary, probiotic Lactic Acid Bacteria exert profound effects on immune system support,
309 benefiting respiratory health by enhancing mucosal immunity and reducing the incidence of
310 infections. Furthermore, their role in allergy management and autoimmune disease modulation
311 highlights their therapeutic potential in maintaining immune balance and overall health. Continued
312 research into their mechanisms and clinical applications is crucial for fully realizing their
313 therapeutic benefits in these areas.

314

315
316 4.3.3. Metabolic Health

317 Probiotic Lactic Acid Bacteria (LAB), including species like Lactobacillus and Bifidobacterium,
318 have emerged as promising agents in promoting metabolic health, particularly in the contexts of
319 obesity, metabolic syndrome, and related disorders. LAB have been investigated for their potential
320 impact on obesity and metabolic syndrome by influencing energy metabolism and adipose tissue
321 function. Research suggests that LAB supplementation may contribute to weight management by
322 modulating energy harvest from the diet and affecting fat accumulation processes within the
323 body(Lin et al., 2022). Furthermore, probiotic LAB have shown beneficial effects in regulating
324 lipid metabolism and glucose homeostasis, essential components in preventing and managing
325 metabolic disorders such as dyslipidemia and diabetes. Studies indicate that LAB supplementation
326 can help improve lipid profiles by reducing total cholesterol and triglyceride levels, while also
327 enhancing glucose metabolism and insulin sensitivity(Xiao et al., 2023). In summary, the
328 therapeutic potential of probiotic Lactic Acid Bacteria extends beyond gastrointestinal health to
329 include significant impacts on metabolic health. Their ability to influence energy metabolism,
330 adipose tissue function, lipid profiles, and glucose homeostasis underscores their role as promising
331 interventions in managing obesity, metabolic syndrome, and related disorders. Continued research
332 is essential to elucidate specific mechanisms and optimize their clinical applications in promoting
333 metabolic well-being.

334 4.3.4. Evidence from clinical studies

335 Clinical studies and meta-analyses have consistently demonstrated the efficacy of probiotic Lactic
336 Acid Bacteria (LAB), including species like Lactobacillus and Bifidobacterium, across a spectrum
337 of health conditions. In terms of gastrointestinal health, probiotic LAB have shown effectiveness
338 in managing diarrhea(McFarland, 2006) highlighted their ability to reduce the severity and
339 duration of antibiotic-associated and infectious diarrhea by restoring gut microbiota balance.
340 Similarly, (Wollny et al., 2021) underscored their role in alleviating symptoms of Irritable Bowel
341 Syndrome (IBS), with certain strains proving beneficial in reducing abdominal pain and improving
342 bowel function.

343 Regarding immune system support, probiotic LAB have been studied for their preventive effects
344 against respiratory infections. Zhao et al. (2023) found that probiotics, particularly strains like
345 Lactobacillus and Bifidobacterium, significantly lowered the incidence and duration of upper
346 respiratory tract infections in both children and adults. In terms of allergy management and
347 autoimmune diseases, Darbandi et al. (2021) highlighted LAB's role in modulating immune
348 responses, which can reduce allergic symptoms and potentially mitigate autoimmune conditions
349 such as atopic dermatitis and rheumatoid arthritis.

350 In the realm of metabolic health, probiotic LAB have shown promise in managing obesity and
351 metabolic syndrome(Ruiz Rodríguez et al., 2022) reviewed trials indicating that specific LAB
352 strains contribute to modest weight loss and improvements in metabolic parameters such as lipid
353 profiles and insulin sensitivity. In addition, Yao et al. (2017) further supported these findings by
354 demonstrating that probiotic supplementation improves lipid metabolism and glucose homeostasis
355 in individuals with type 2 diabetes. These studies collectively underscore the diverse therapeutic
356 potential of probiotic Lactic Acid Bacteria in promoting health across different systems. From
357 gastrointestinal benefits to immune system modulation and metabolic improvements, probiotics
358 offer multifaceted benefits supported by clinical evidence. However, variations in study designs,
359 populations, probiotic strains, and dosages suggest the need for tailored approaches in clinical
360 practice. Continued research is essential to optimize the use of probiotics and further elucidate
361 their mechanisms of action for targeted therapeutic applications.

362 Futures perspective

363 The future of probiotic Lactic Acid Bacteria promises advancements in personalized medicine,
364 expanded clinical applications, innovative delivery systems, robust regulatory frameworks, and
365 integration with digital health solutions. These developments will elevate probiotics from dietary
366 supplements to integral components of precision healthcare, offering targeted therapies for
367 enhancing gut health, immune function, and metabolic well-being across diverse patient
368 populations. Continued interdisciplinary research and collaboration will drive these advancements,
369 paving the way for a new era in probiotic-based therapies and preventive health strategies.

370

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Declaration of Interest

The authors have no conflict of interest