Search results for: Lactobacillus salivarus

Authors: Thalía Moreno-García Malo, Santiago Torres-Ríos, María G. González-Cruz, María M. Hernández-Arroyo, Sergio R. Trejo-Estrada

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Agave atrovirens is the main source of agave sap, the raw material for the production of pulque, an artisanal fermented beverage, traditional since prehispanic times in the highlands of central Mexico. Agave sap is rich in glucose, sucrose and fructooligosaccharides, and strongly differs from agave syrup from A. tequilana, which is mostly a high molecular weight fructan. Agave sap is converted into pulque by a highly diverse microbial community which includes bacteria, yeast and even filamentous fungi. The bacterial diversity has been recently studied. But the composition of consortia derived from directed enrichments differs sharply from the whole fermentative consortium. Using classical microbiology methods, and selective liquid and solid media formulations, either bacterial or fungal consortia were developed and analyzed. Bacterial consortia able to catabolize specific prebiotic saccharides were selected and preserved for future developments. Different media formulations, selective for bacterial genera such as Bifidobacterium, Lactobacillus, Pediococcus, Lactococcus and Enterococcus were also used. For yeast, specific media, osmotic pressure and unique carbon sources were used as selective agents. Results show that most groups are represented in the enrichment cultures; although very few are recoverable from the whole consortium in artisanal pulque. Diversity and abundance vary among consortia. Potential bacterial probiotics obtained from agave sap and agave juices show tolerance to hydrochloric acid, as well as strong antimicrobial activity.

Keywords: Agave, pulque, microbial consortia, prebiotic activity

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Authors: Hanan Al-Khalaifah, Afaf Al-Nasser

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Probiotics and prebiotics claimed to serve as effective alternatives to antibiotics in the poultry. This study aims to investigate the effect of different probiotics and prebiotics on the economic impact analysis of the use of probiotics and prebiotics in broiler feed. The study involved four broiler cycles, two during winter and two during summer. In the first two cycles (summer and winter), different types of prebiotics and probiotics were used. The probiotics were Bacillus coagulans (1 g/kg dried culture) and Lactobacillus (1 g/kg dried culture of 12 commercial strains), and prebiotics included fructo-oligosaccharides (FOS) (5 g/kg) and mannan-oligosaccharide (MOS) derived from Saccharomyces cerevisiae (5 g/kg). Based on the results obtained, the best treatment was chosen to be FOS, from which different ratios were used in the last two cycles during winter and summer. The levels of FOS chosen were 0.3, 0.5, and 0.7% of the diet. From an economic point of view, it was generally concluded that in all dietary treatments, food was consumed less in cycle 1 than in cycle 2, the total body weight gain was more in cycle 1 than cycle 2, and the average feed efficiency was less in cycle l than cycle 2. This indicates that the weather condition affected better in cycle 1. Also, there were very small differences between the dietary treatments in each cycle. In cycle 1, the best total feed consumption was for the FOS treatment, the highest total body weight gain and average feed efficiency were for B. coagulans. In cycle 2, all performance was better in FOS treatment. FOS significantly reduced the Salmonella sp. counts in the intestine, where the environment was driven towards acidity. FOS was the best on the average taste panel study of the produced meat. Accordingly, FOS prebiotic was chosen to be the best treatment to be used in cycles 3 and 4. The economic impact analysis generally revealed that there were no big differences between the treatments in all of the studied indicators, but there was a difference between the cycles.

Keywords: antibiotic, economic impact, prebiotic, probiotic, broiler

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Authors: Minh-Phuong Ngoc Bui, Abdennour Abbas

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Microbial spoilage of food/drug has been a constant nuisance and an unavoidable problem throughout history that affects food/drug quality and safety in a variety of ways. A simple and rapid test of fungi and bacteria in food/drugs and environmental clinical samples is essential for proper management of contamination. A number of different techniques have been developed for detection and enumeration of foodborne microorganism including plate counting, enzyme-linked immunosorbent assay (ELISA), polymer chain reaction (PCR), nucleic acid sensor, electrical and microscopy methods. However, the significant drawbacks of these techniques are highly demand of operation skills and the time and cost involved. In this report, we introduce a rapid method for detection of bacteria and fungi in food/drug products using a specific interaction between a reducing agent (tris(2-carboxylethyl)phosphine (TCEP)) and the microbial surface proteins. The chemical reaction was transferred to a transduction system using gold nanoplates-enhanced chemiluminescence. We have optimized our nanoplates synthetic conditions, characterized the chemiluminescence parameters and optimized conditions for the microbial assay. The new detection method was applied for rapid detection of bacteria (E.coli sp. and Lactobacillus sp.) and fungi (Mucor sp.), with limit of detection as low as single digit cells per mL within 10 min using a portable luminometer. We expect our simple and rapid detection method to be a powerful alternative to the conventional plate counting and immunoassay methods for rapid screening of microorganisms in food/drug products.

Keywords: microorganism testing, gold nanoplates, chemiluminescence, reducing agents, luminol

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Authors: Nurul Aqilah Mohd Zaini, Afroditi Chatzifragkou, Dimitris Charalampopoulos

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D-Lactic acid production is gaining increasing attention due to the thermostable properties of its polymer, Polylactic Acid (PLA). In this study, D-lactic acid was produced in microbial cultures using Lactobacillus coryniformis subsp. torquens as D-lactic acid producer and hydrolysates of Dried Distillers Grains with Solubles (DDGS) as fermentation substrate. Prior to fermentation, DDGS was first alkaline pretreated with 5% (w/v) NaOH, for 15 minutes (121oC/ ~16 psi). This led to the generation of DDGS solid residues, rich in carbohydrates and especially cellulose (~52%). The carbohydrate-rich solids were then subjected to enzymatic hydrolysis with Accellerase® 1500. For Separate Hydrolysis and Fermentation (SHF), enzymatic hydrolysis was carried out at 50oC for 24 hours, followed by fermentation of D-lactic acid at 37oC in controlled pH 6. The obtained hydrolysate contained 24 g/l glucose, 5.4 g/l xylose and 0.6 g/l arabinose. In the case of Simultaneous Saccharification and Fermentation (SSF), hydrolysis and fermentation were conducted in a single step process at 37oC in pH 5. The enzymatic hydrolysis of DGGS pretreated solids took place mostly during lag phase of L. coryniformis fermentation, with only a small amount of glucose consumed during the first 6 h. When exponential phase was started, glucose generation reduced as the microorganism started to consume glucose for D-lactic acid production. Higher concentrations of D-lactic acid were produced when SSF approach was applied, with 28 g/l D-lactic acid after 24 h of fermentation (84.5% yield). In contrast, 21.2 g/l D-lactic acid were produced when SHF was used. The optical pu rity of D-lactic acid produced from both experiments was 99.9%. Besides, approximately 2 g/l acetic acid was also generated due to lactic acid degradation after glucose depletion in SHF. SSF was proved an efficient towards DDGS ulilisation and D-lactic acid production, by reducing the overall processing time, yielding sufficient D-lactic acid concentrations without the generation of fermentation by-products.

Keywords: DDGS, alkaline pretreatment, SSF, D-lactic acid

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Authors: Samuel E. Okere, Anthony E. Ataga

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Introduction: The microbial ecology of Pleurotus osteratus and Pleurotus tuber–regium spent mushroom substrate (SMS) were characterized to determine other ways of its utilization. Materials and Methods: The microbiological properties of the spent mushroom substrate were determined using standard methods. This study was carried out at the Microbiology Laboratory University of Port Harcourt, Rivers State, Nigeria. Results: Quantitative microbiological analysis revealed that Pleurotus osteratus spent mushroom substrate (POSMS) contained 7.9x10⁵ and 1.2 x10³ cfu/g of total heterotrophic bacteria and total fungi count respectively while Pleurotus tuber-regium spent mushroom substrate (PTSMS) contained 1.38x10⁶ and 9.0 x10² cfu/g of total heterotrophic bacteria count and total fungi count respectively. The fungi species encountered from Pleurotus tuber-regium spent mushroom substrate (PTSMS) include Aspergillus and Cladosporum species, while Aspergillus and Penicillium species were encountered from Pleurotus osteratus spent mushroom substrate (POSMS). However, the bacteria species encountered from Pleurotus tuber-regium spent mushroom substrate include Bacillus, Acinetobacter, Alcaligenes, Actinobacter, and Pseudomonas species while Bacillus, Actinobacteria, Aeromonas, Lactobacillus and Aerococcus species were encountered from Pleurotus osteratus spent mushroom substrate (POSMS). Conclusion: Therefore based on the findings from this study, it can be concluded that spent mushroom substrate contain microorganisms that can be utilized both in bioremediation of oil-polluted soils as they contain important hydrocarbon utilizing microorganisms such as Penicillium, Aspergillus and Bacillus species and also as sources of plant growth-promoting rhizobacteria (PGPR) such as Pseudomonas and Bacillus species which can induce resistance on plants. However, further studies are recommended, especially to molecularly characterize these microorganisms.

Keywords: characterization, microorganisms, mushroom, spent substrate

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Authors: Mohammad Aladwan, Adelia Skripova

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Bioremediation aspects of crude oil-polluted fields can be achieved by isolation and identification of bacterial species from oil-contaminated soil in order to choose the most active isolates and increase the strength of others. In this study, oil-degrading bacteria were isolated and identified from oil-contaminated soil samples in northeastern Jordan. The bacterial growth count (CFU/g) was between 1.06×10⁵ and 0.75×10⁹. Eighty-two bacterial isolates were characterized by their morphology and biochemical tests. The identified bacterial genera included: Klebsiella, Staphylococcus, Citrobacter, Lactobacillus, Alcaligenes, Pseudomonas, Hafnia, Micrococcus, Rhodococcus, Serratia, Enterobacter, Bacillus, Salmonella, Mycobacterium, Corynebacterium, and Acetobacter. Molecular identification of a universal primer 16S rDNA gene was used to identify four bacterial isolates: Microbacterium esteraromaticum strain L20, Pseudomonas stutzeri strain 13636M, Klebsilla pneumoniae, and uncultured Klebsilla sp., known as new strains. Our results indicate that their specific oil-degrading bacteria isolates might have a high strength of oil degradation from oil-contaminated sites. Staphylococcus intermedius (75%), Corynebacterium xerosis (75%), and Pseudomonas fluorescens (50%) showed a high growth rate on different types of hydrocarbons, such as crude oil, toluene, naphthalene, and hexane. In addition, monooxygenase and catechol 2,3-dioxygenase were detected in 17 bacterial isolates, indicating their superior hydrocarbon degradation potential. Total petroleum hydrocarbons were analyzed using gas chromatography for soil samples. Soil samples M5, M7, and M8 showed the highest levels (43,645, 47,805, and 45,991 ppm, respectively), and M4 had the lowest level (7,514 ppm). All soil samples were analyzed for heavy metal contamination (Cu, Cd, Mn, Zn, and Pb). Site M7 contains the highest levels of Cu, Mn, and Pb, while Site M8 contains the highest levels of Mn and Zn. In the future, these isolates of bacteria can be used for the cleanup of oil-contaminated soil.

Keywords: bioremediation, 16S rDNA gene, oil-degrading bacteria, hydrocarbons

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Authors: Alloysius C. Ogodo, Ositadinma C. Ugbogu, Uzochukwu G. Ekeleme

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Food poisoning and infection by bacteria are of public health significance to both developing and developed countries. Samples of ogi (akamu) prepared from white and yellow variety of maize sold in Uturu and Okigwe were analyzed together with the laboratory prepared ogi for microbial quality using the standard microbiological methods. The analyses showed that both white and yellow variety had total bacterial counts (cfu/g) of 4.0 ×107 and 3.9 x 107 for the laboratory prepared ogi while the commercial ogi had 5.2 x 107 and 4.9 x107, 4.9 x107 and 4.5 x107, 5.4 x107 and 5.0 x107 for Eke-Okigwe, Up-gate and Nkwo-Achara market respectively. The Staphylococcal counts ranged from 2.0 x 102 to 5.0 x102 and 1.0 x 102 to 4.0 x102 for the white and yellow variety from the different markets while Staphylococcal growth was not recorded on the laboratory prepared ogi. The laboratory prepared ogi had no Coliform growth while the commercially prepared ogi had counts of 0.5 x103 to 1.6 x 103 for white variety and 0.3 x 103 to 1.1 x103 for yellow variety respectively. The Lactic acid bacterial count of 3.5x106 and 3.0x106 was recorded for the laboratory ogi while the commercially prepared ogi ranged from 3.2x106 to 4.2x106 (white variety) and 3.0 x106 to 3.9 x106 (yellow). The presence of bacteria isolates from the commercial and laboratory fermented ogi showed that Lactobacillus sp, Leuconostoc sp and Citrobacter sp were present in all the samples, Micrococcus sp and Klebsiella sp were isolated from Eke-Okigwe and ABSU-up-gate markets varieties respectively, E. coli and Staphylococcus sp were present in Eke-Okigwe and Nkwo-Achara markets while Salmonella sp were isolated from the three markets. Hence, there are chances of contracting food borne diseases from commercially prepared ogi. Therefore, there is the need for sanitary measures in the production of fermented cereals so as to minimize the rate of food borne pathogens during processing and storage.

Keywords: ogi, fermentation, bacterial quality, lactic acid bacteria, maize

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Authors: Genesis Julyus T. Agcaoili, Esperanza C. Cabrera

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Thirty (30) isolates of lactic acid bacteria (LAB) from traditionally-prepared fermented products specifically fermented soy-bean paste, fermented mustard and fermented rice-fish mixture were studied for their in vitro antimicrobial and cytotoxic activities. Seventeen (17) isolates were identified as Lactobacillus plantarum, while 13 isolates were identified as Enterococcus spp using 16s rDNA sequences. Disc diffusion method was used to determine the antibacterial activity of LAB against Staphylococcus aureus (ATCC 25923) and Escherichia coli (ATCC 25922), while the modified agar overlay method was used to determine the antifungal activity of LAB isolates on the yeast Candida albicans, and the dermatophytes Microsporum gypseum, Trichophyton rubrum and Epidermophyton floccosum. The filter-sterilized LAB supernatants were evaluated for their cytotoxicity to mammalian colon cancer cell lines (HT-29 and HCT116) and normal human dermal fibrolasts (HDFn) using resazurin assay (PrestoBlueTM). Colchicine was the positive control. No antimicrobial activity was observed against the bacterial test organisms and the yeast Candida albicans. On the other hand, all of the tested LAB strains were fungicidal for all the test dermatophytes. Cytotoxicity index profiles of the supernatants of the 15 randomly picked LABs and negative control (brain heart infussion broth) suggest nontoxicity to the cells when compared to colchicine, whereas all LAB supernatants were found to be cytotoxic to HT-29 and HCT116 colon cancer cell lines. Results provide strong support for the role of the lactic acid bacteria studied in antimicrobial treatment and anticancer therapy.

Keywords: antimicrobial, fermented products, fungicidal activity, lactic acid bacteria, probiotics

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Authors: Henry O. Udeha, Kwaku G. Duodub, Afam I. O. Jideanic

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Finger millet (FM) [Eleusine coracana] is considered as a potential ‘‘super grain’’ by the United States National Academies as one of the most nutritious among all the major cereals. The regular consumption of FM-based diets has been associated with reduced risk of diabetes, cataract and gastrointestinal tract disorder. Hyperglycaemic, hypocholesterolaemic and anticataractogenic, and other health improvement properties have been reported. This study examined the effect of fermentation time and microbial source on physicochemical properties, phenolic compounds and antioxidant activity of two finger millet (FM) malt flours. Sorghum was used as an external reference. The grains were malted, mashed and fermented using the grain microflora and Lactobacillus fermentum. The phenolic compounds of the resulting beverage were identified and quantified using ultra-performance liquid chromatography (UPLC) and mass spectrometer system (MS). A fermentation-time dependent decrease in pH and viscosities of the beverages, with a corresponding increase in sugar content were noted. The phenolic compounds found in the FM beverages were protocatechuic acid, catechin and epicatechin. Decrease in total phenolics of the beverages was observed with increased fermentation time. The beverages exhibited 2, 2-diphenyl-1-picrylhydrazyl, 2, 2՛-azinobis-3-ethylbenzthiazoline-6-sulfonic acid radical scavenging action and iron reducing activities, which were significantly (p < 0.05) reduced at 96 h fermentation for both microbial sources. The 24 h fermented beverages retained a higher amount of total phenolics and had higher antioxidant activity compared to other fermentation periods. The study demonstrates that FM could be utilised as a functional grain in the production of non-alcoholic beverage with important phenolic compounds for health promotion and wellness.

Keywords: antioxidant activity, eleusine coracana, fermentation, phenolic compounds

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Authors: Shilpi Malik, Ramneek Kaur, Archita Gupta, Deepshikha Yadav, Ashwani Mathur, Manisha Singh

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Glucosamine (GS) is the most abundant naturally occurring amino monosaccharide and is normally produced in human body via cellular glucose metabolism. It is regarded as the building block of cartilage matrix and is also an essential component of cartilage matrix repair mechanism. Besides that, it can also be explored for its prebiotic potential as many bacterial species are known to utilize the amino sugar by acquiring them to form peptidoglycans and lipopolysaccharides in the bacterial cell wall. Glucosamine can therefore be considered for its fermentation by bacterial species present in the gut. Current study is focused on exploring the potential of glucosamine as prebiotic. The studies were done to optimize considerable concentration of GS to reach GI tract and being fermented by the complex gut microbiota and food grade GS was added to various Simulated Fluids of Gastro-Intestinal Tract (GIT) such as Simulated Saliva, Gastric Fluid (Fast and Fed State), Colonic fluid, etc. to detect its degradation. Since it was showing increase in microbial growth (CFU) with time, GS was Further, encapsulated to increase its residential time in the gut, which exhibited improved resistance to the simulated Gut conditions. Moreover, prepared microspehres were optimized and characterized for their encapsulation efficiency and toxicity. To further substantiate the prebiotic activity of Glucosamine, studies were also performed to determine the effect of Glucosamine on the known probiotic bacterial species, i.e. Lactobacillus delbrueckii (MTCC 911) and Bifidobacteriumbifidum (MTCC 5398). Culture conditions for glucosamine will be added in MRS media in anaerobic tube at 0.20%, 0.40%, 0.60%, 0.80%, and 1.0%, respectively. MRS media without GS was included in this experiment as the control. All samples were autoclaved at 118° C for 15 min. Active culture was added at 5% (v/v) to each anaerobic tube after cooling to room temperature and incubated at 37° C then determined biomass and pH and viable count at incubation 18h. The experiment was completed in triplicate and the results were presented as Mean ± SE (Standard error).The experimental results are conclusive and suggest Glucosamine to hold prebiotic properties.

Keywords: gastro intestinal tract, microspheres, peptidoglycans, simulated fluid

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Authors: Ziaul Islam, Asad Sultan, Sarzamin Khan

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Organic acids and micro encapsulated phyto essential oils have revealed great potential as an antibiotic replacement and as an additive to work tremendously for the health maintenance of broiler chicken. To explore more about organic acids, a total of 600 day-old broiler chicks (Cobb-500) were procured from a local hatchery and distributed into 5 treatment groups having 6 replicates of 20 birds each; the duration of the biological trial was of 35 days. Group T1 served as a control group that were fed on corn soy-based diet only. T2 were fed with a diet having 6% poultry by-product meal (PBM) diet, T3, T4, and T5 were served as the same diet as T2 but supplemented with an organic acid, phyto essential oils alone, and a combination, respectively. The findings declared significant improvement (p<0.05) in body weight gain and FCR in groups T3, T4, and T5 while feed intake was not affected. European broiler performance indicators like production efficiency factor (EPEF) and broiler index (EBI) were improved significantly (p<0.05) by the treatments T3, T4, and T5 compared with T1 and T2. Carcass evaluation depicted significantly better (p<0.05) dressed and eviscerated weight along with carcass yield (T3, T4, T5). Broilers fed organic acid and phyto essential oils supplemented diet had significantly lower (p<0.05) Clostridium perfringens, Escherichia coliand Salmonella and increased Lactobacillus counts. Likewise, antibody titer against ND, IB, and IBD were also significantly (p<0.05) improved by the treatments T3, T4 and T5compared with the T1and T2. Litter moisture content was significantly (p<0.05) reduced by treatmentsT3, T4, and T5 on day 28 and 35 compared with the T1 and T2. These findings of the present study revealed that supplementation of organic acids blend and phyto-essential oils as an as an substitute to improve the performance of broilers without the use of feed antibiotics in broilers fed with 6% poultry by-product meal based diet.

Keywords: organic acid, phyto essential oils, growth performance, PBM, gut health, microbiota, immunity

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Authors: Muhammad Adeel Arshad, Shaukat Ali Bhatti, Faiz-ul Hassan

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Manipulating microbial ecosystem in the rumen is considered as an important strategy to optimize production efficiency in ruminants. In the past, antibiotics and synthetic chemical compounds have been used for the manipulation of rumen fermentation. However, since the non-therapeutic use of antibiotics has been banned, efforts are being focused to search out safe alternative products. In tropics, crop residues and forage grazing are major dietary sources for ruminants. Poor digestibility and utilization of these feedstuffs by animals is a limiting factor to exploit the full potential of ruminants in this area. Hence, there is a need to enhance the utilization of these available feeding resources. One of the potential strategies in this regard is the use of direct-fed microbes. Bacteria and fungi are mostly used as direct-fed microbes to improve animal health and productivity. Commonly used bacterial species include lactic acid-producing and utilizing bacteria (Lactobacillus, Streptococcus, Enterococcus, Bifidobacterium, and Bacillus) and fungal species of yeast are Saccharomyces and Aspergillus. Direct-fed microbes modulate microbial balance in the gastrointestinal tract through the competitive exclusion of pathogenic species and favoring beneficial microbes. Improvement in weight gain and feed efficiency has been observed as a result of feeding direct-fed bacteria. The use of fungi as a direct-fed microbe may prevent excessive production of lactate and harmful oxygen in the rumen leading to better feed digestibility. However, the mechanistic mode of action for bacterial or fungal direct-fed microbes has not been established yet. Various reports have confirmed an increase in dry matter intake, milk yield, and milk contents in response to the administration of direct-fed microbes. However, the application of a direct-fed microbe has shown variable responses mainly attributed to dosages and strains of microbes. Nonetheless, it is concluded that the inclusion of direct-fed microbes may mediate the rumen ecosystem to manage lactic acid production and utilization in both clinical and sub-acute rumen acidosis.

Keywords: microbes, roughages, rumen, feed efficiency, production, fermentation

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Authors: Amina Aloui, Jalila Ben Salah-Abbès, Abdellah Zinedine, Amar Riba, Noel Durand, Catherine Brabet, Didier Montet, Samir Abbès

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Mycotoxin contamination of food and feed-isa globaconcern, both economically and for public health. Aflatoxin M1 (AFM1) is the principal hydroxylated metabolite of aflatoxin B1. It is frequently found in milk and other dairy products. It is responsible for the development of hepatocellular carcinoma and immunotoxic in humans and animals. The reduction of its bioavailabilitybecomesa great demand in order to protect human and animal health. The use of probiotic bacteria and clay are demonstrated to be able to bind AFM1 in vitro. This study aimed to investigate, in vivo, the activity of two-component mixture: L. rhamnosusGAF06 (LR) and bentonite for reducing the oxidative stress and the histological alterationsinduced by AFM1 in the liver andkidneys. For the experiment, male mice were divided into 7 groups (6 mice/group) and treated, orally, by AFM1, alone or in combination with LR and/or bentonite, for 10 days as follows: group 1 control, group 2 treated with LR alone (2.108 CFU/ml), group 3 treated with bentonite alone (1g/kg), group 4 treated with AFM1 alone (100μg/kg), group 5 co-treated with LR+AFM1, group 6 co-treated with bentonite+AFM1, group 7 co-treated with bentonite+LR+AFM1. At the end of the treatment, the mice were sacrificed, and the livers and kidneys were collected for histological assays. Intracellular antioxidant activities and lipid peroxidation were also studied. The results showed that AFM1causeddamage in liver and kidney tissues, being evidence of hepatotoxicity and nephrotoxicity marked by necrotic cells. It increased the MDA level and decreased the antioxidant enzyme activities (SOD) in both organs. In contrast, the co-treatment with AFM1 plus LR and/or bentonitesignificantly improved the hepatic and renal tissues, regulated kidney, and liver antioxidant enzyme activities. This improvement was more remarkable with the administration of LR-bentonite mixture with AFM1.LR and bentonite alone showed to be safe during the treatment. This mixture can be a promising candidate for future applications in biotechnological processes that aimed to detoxify AFM1in food and feed.

Keywords: aflatoxin M1, bentonite, L. rhamnosus GAF06, oxidative stress, prevention

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Authors: Zelal Polat, Şebnem Harsa, Semra Ülkü

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Lactic acid exists in nature optically in two forms, L(+), D(-)-lactic acid, and has been used in food, leather, textile, pharmaceutical and cosmetic industries. Moreover, L(+)-lactic acid constitutes the raw material for the production of poly-L-lactic acid which is used in biomedical applications. Microbially produced lactic acid was aimed to be recovered from the fermentation media efficiently and economically. Among the various downstream operations, ion exchange chromatography is highly selective and yields a low cost product recovery within a short period of time. In this project, Lactobacillus casei NRRL B-441 was used for the production of L(+)-lactic acid from whey by fermentation at pH 5.5 and 37°C that took 12 hours. The product concentration was 50 g/l with 100% L(+)-lactic acid content. Next, the suitable resin was selected due to its high sorption capacity with rapid equilibrium behavior. Dowex marathon WBA, weakly basic anion exchanger in OH form reached the equilibrium in 15 minutes. The batch adsorption experiments were done approximately at pH 7.0 and 30°C and sampling was continued for 20 hours. Furthermore, the effect of temperature and pH was investigated and their influence was found to be unimportant. All the adsorption/desorption experiments were applied to both model lactic acid and biomass free fermentation broth. The ion exchange equilibria of lactic acid and L(+)-lactic acid in fermentation broth on Dowex marathon WBA was explained by Langmuir isotherm. The maximum exchange capacity (qm) for model lactic acid was 0.25 g La/g wet resin and for fermentation broth 0.04 g La/g wet resin. The equilibrium loading and exchange efficiency of L(+)-lactic acid in fermentation broth were reduced as a result of competition by other ionic species. The competing ions inhibit the binding of L(+)-lactic acid to the free sites of ion exchanger. Moreover, column operations were applied to recover adsorbed lactic acid from the ion exchanger. 2.0 M HCl was the suitable eluting agent to recover the bound L(+)-lactic acid with a flowrate of 1 ml/min at ambient temperature. About 95% of bound L(+)-lactic acid was recovered from Dowex marathon WBA. The equilibrium was reached within 15 minutes. The aim of this project was to investigate the purification of L(+)-lactic acid with ion exchange method from fermentation broth. The additional goals were to investigate the end product purity, to obtain new data on the adsorption/desorption behaviours of lactic acid and applicability of the system in industrial usage.

Keywords: fermentation, ion exchange, lactic acid, purification, whey

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Authors: Sergio D’Ambrosioa, Azza Dobousa, Chiara Schiraldia, Donatella Ciminib

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Probiotics are living microorganisms that give beneficial effects while consumed. Lactic acid bacteria and bifidobacteria are among the most representative strains assessed as probiotics and exploited as food supplements. Numerous studies demonstrated their potential as a therapeutic candidate for a variety of diseases (restoring gut flora, lowering cholesterol, immune response-enhancing, anti-inflammation and anti-oxidation activities). These beneficial actions are also due to biomolecules produced by probiotics, such as exopolysaccharides (EPSs), that demonstrate plenty of beneficial properties such as antimicrobial, antitumor, anti-biofilm, antiviral and immunomodulatory activities. Limosilactobacillus fermentum is a widely studied member of probiotics; however, few data are available on the development of fermentation and downstream processes for the production of viable biomasses for potential industrial applications. However, few data are available on the development of fermentation processes for the large-scale production of probiotics biomass for industrial applications and for purification processes of EPSs at an industrial scale. For this purpose, L. fermentum strain was isolated from buffalo milk and used as a test example for biotechnological process development. The strain was able to produce up to 109 CFU/mL on a (glucose-based) semi-defined medium deprived of animal-derived raw materials up to the pilot scale (150 L), demonstrating improved results compared to commonly used, although industrially not suitable, media-rich of casein and beef extract. Biomass concentration via microfiltration on hollow fibers, and subsequent spray-drying allowed to recover of about 5.7 × 1010CFU/gpowder of viable cells, indicating strain resistance to harsh processing conditions. Overall, these data demonstrate the possibility of obtaining and maintaining adequate levels of viable L. fermentum cells by using a simple approach that is potentially suitable for industrial development. A downstream EPS purification protocol based on ultrafiltration, precipitation and activated charcoal treatments showed a purity of the recovered polysaccharides of about 70-80%.

Keywords: probiotics, fermentation, exopolysaccharides (EPSs), purification

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Authors: Deeba N. Baig, Ateeqa Ijaz, Saloome Rafiq

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Camel is a religious and culturally significant animal in Asian and African regions. In Pakistan Dromedary and Bactrian are common camel breeds. Other than the transportation use, it is a pivotal source of milk and meat. The quality of its milk and meat is predominantly dependent on the geographical location and variety of vegetation available for the diet. Camel milk (CM) is highly nutritious because of its reduced cholesterol and sugar contents along with enhanced minerals and vitamins level. The absence of beta-lactoglobulin (like human milk), makes CM a safer alternative for infants and children having Cow Milk Allergy (CMA). In addition to this, it has a unique probiotic profile both in raw and fermented form. Number of Lactic acid bacteria (LAB) including lactococcus, lactobacillus, enterococcus, streptococcus, weissella, pediococcus and many other bacteria have been detected. From these LAB Lactobacilli, Bifidobacterium and Enterococcus are widely used commercially for fermentation purpose. CM has high therapeutic value as its effectiveness is known against various ailments like fever, arthritis, asthma, gastritis, hepatitis, Jaundice, constipation, postpartum care of women, anti-venom, dropsy etc. It also has anti-diabetic, anti-microbial, antitumor potential along with its robust efficacy in the treatment of auto-immune disorders. Recently, the role of CM has been explored in brain-gut axis for the therapeutics of neurodevelopmental disorders. In this connection, a lot of grey area was available to explore the probiotics and therapeutics latent in the CM available in Pakistan. Thus, current study was designed to explore the predominant probiotic flora and antimicrobial potential of CM from different local breeds of Pakistan. The probiotics have been identified through biochemical, physiological and ribo-typing methods. In addition to this, bacteriocins (antimicrobial-agents) were screened through PCR-based approach. Results of this study revealed that CM from different breeds of camel depicted a number of similar probiotic candidates along with the range of limited variability. However, the nucleotide sequence analysis of selected anti-listerial bacteriocins exposed least variability. As a conclusion, the CM has sufficient probiotic availability and significant anti-microbial potential.

Keywords: bacteriocins, camel milk, probiotics potential, therapeutics

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Authors: Regiane A. Oliveira, Carlos E. Vaz Rossell, Rubens Maciel Filho

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Lactic acid, besides a valuable chemical may be considered a platform for other chemicals. In fact, the feasibility of hemicellulosic sugars as feedstock for lactic acid production process, may represent the drop of some of the barriers for the second generation bioproducts, especially bearing in mind the 5-carbon sugars from the pre-treatment of sugarcane bagasse. Bearing this in mind, the purpose of this study was to use the hemicellulosic liquor from sugarcane bagasse as a substrate to produce lactic acid by fermentation. To release of sugars from hemicellulose it was made a pre-treatment with a diluted sulfuric acid in order to obtain a xylose's rich liquor with low concentration of inhibiting compounds for fermentation (≈ 67% of xylose, ≈ 21% of glucose, ≈ 10% of cellobiose and arabinose, and around 1% of inhibiting compounds as furfural, hydroxymethilfurfural and acetic acid). The hemicellulosic sugars associated with 20 g/L of yeast extract were used in a fermentation process with Lactobacillus plantarum to produce lactic acid. The fermentation process pH was controlled with automatic injection of Ca(OH)2 to keep pH at 6.00. The lactic acid concentration remained stable from the time when the glucose was depleted (48 hours of fermentation), with no further production. While lactic acid is produced occurs the concomitant consumption of xylose and glucose. The yield of fermentation was 0.933 g lactic acid /g sugars. Besides, it was not detected the presence of by-products, what allows considering that the microorganism uses a homolactic fermentation to produce its own energy using pentose-phosphate pathway. Through facultative heterofermentative metabolism the bacteria consume pentose, as is the case of L. plantarum, but the energy efficiency for the cell is lower than during the hexose consumption. This implies both in a slower cell growth, as in a reduction in lactic acid productivity compared with the use of hexose. Also, L. plantarum had shown to have a capacity for lactic acid production from hemicellulosic hydrolysate without detoxification, which is very attractive in terms of robustness for an industrial process. Xylose from hydrolyzed bagasse and without detoxification is consumed, although the hydrolyzed bagasse inhibitors (especially aromatic inhibitors) affect productivity and yield of lactic acid. The use of sugars and the lack of need for detoxification of the C5 liquor from sugarcane bagasse hydrolyzed is a crucial factor for the economic viability of second generation processes. Taking this information into account, the production of second generation lactic acid using sugars from hemicellulose appears to be a good alternative to the complete utilization of sugarcane plant, directing molasses and cellulosic carbohydrates to produce 2G-ethanol, and hemicellulosic carbohydrates to produce 2G-lactic acid.

Keywords: fermentation, lactic acid, hemicellulosic sugars, sugarcane

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Authors: Komal Khan, Hafsa Zaneb, Saima Masood, Muhammad Younus, Sanan Raza

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Cigarette smoke induces many physiological and pathological changes in respiratory tract like goblet cell hyperplasia and regional distention of airspaces. It is also associated with elevation of inflammatory profiles in different airway compartments. As probiotics are generally known to promote mucosal tolerance, it was postulated that prophylactic use of probiotics can be helpful in reduction of respiratory damage induced by cigarette smoke exposure. Twenty-four adult mice were randomly divided into three groups (cigarette-smoke (CS) group, cigarette-smoke+ Lactobacillus (CS+ P) group, control (Cn) group), each having 8 mice. They were exposed to cigarette smoke for 28 days (6 cigarettes/ day for 6 days/week). Wright-Giemsa staining of bronchoalveolar lavage fluid (BALF) was performed in three mice per group. Tissue samples of trachea and lungs of 7 mice from each group were processed by paraffin embedding technique for haematoxylin & eosin (H & E) and alcian blue- periodic acid-Schiff (AB-PAS) staining. Then trachea (goblet cell number, ratio and loss of cilia) and lungs (airspace distention) were studied. The results showed that the number of goblet cells was increased in CS group as a result of defensive mechanism of the respiratory system against irritating substances. This study also revealed that the cells of CS group having acidic glycoprotein were found to be higher in quantity as compared to those containing neutral glycoprotein. However, CS + P group showed a decrease in goblet cell index due to enhanced immunity by prophylactically used probiotics. Moreover, H & E stained tracheas showed significant loss of cilia in CS group due to propelling of mucous but little loss in CS + P group because of having good protective tracheal epithelium. In lungs, protection of airspaces was also much more evident in CS+ P group as compared to CS group having distended airspaces, especially at 150um distance from terminal bronchiole. In addition, a comprehensive analysis of inflammatory cells population of BALF showed neutrophilia and eosinophilia was significantly reduced in CS+ P group. This study proved that probiotics are found to be useful for reduction of changes in micro-architecture of the respiratory system. Thus, dietary supplementation of probiotic as prophylactic measure can be useful in achieving immunomodulatory effects.

Keywords: cigarette smoke, probiotics, goblet cells, airspace enlargement, BALF

Procedia PDF Downloads 364

Authors: J. M. Cruz, X. Vecıno, L. Rodrıguez-López, J. M. Dominguez, A. B. Moldes

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The cosmetic and personal care industry are the fields where biosurfactants could have more possibilities of success because in this kind of products the replacement of synthetic detergents by natural surfactants will provide an additional added value to the product, at the same time that the harmful effects produced by some synthetic surfactants could be avoided or reduced. Therefore, nowadays, consumers are disposed to pay and additional cost if they obtain more natural products. In this work we provide data about the potential of biosurfactants in the cosmetic and personal care industry. Biosurfactants from corn steep liquor, that is a fermented and condensed stream, have showed good surface-active properties, reducing substantially the surface tension of water. The bacteria that usually growth in corn steep liquor comprises Lactobacillus species, generally recognize as safe. The biosurfactant extracted from CSL consists of a lipopeptide, composed by fatty acids, which can reduce the surface tension of water in more than 30 units. It is a yellow and viscous liquid with a density of 1.053 mg/mL and pH=4. By these properties, they could be introduced in the formulation of cosmetic creams, hair conditioners or shampoos. Moreover this biosurfactant extracted from corn steep liquor, have showed a potent antimicrobial effect on different strains of Streptococcus. Some species of Streptococcus are commonly found weakly living in the human respiratory and genitourinary systems, producing several diseases in humans, including skin diseases. For instance, Streptococcus pyogenes produces many toxins and enzymes that help to stabilize skin infections; probably biosurfactants from corn steep liquor can inhibit the mechanisms of the S. pyogenes enzymes. S. pyogenes is an important cause of pharyngitis, impetigo, cellulitis and necrotizing fasciitis. In this work it was observed that 50 mg/L of biosurfactant extract obtained from corn steep liquor is able to inhibit more than 50% the growth of S. pyogenes. Thus, cosmetic and personal care products, formulated with biosurfactants from corn steep liquor, could have prebiotic properties. The natural biosurfactant presented in this work and obtained from corn milling industry streams, have showed a high potential to provide an interesting and sustainable alternative to those, antibacterial and surfactant ingredients used in cosmetic and personal care manufacture, obtained by chemical synthesis, which can cause irritation, and often only show short time effects.

Keywords: antimicrobial activity, biosurfactants, cosmetic, personal care

Procedia PDF Downloads 257

Authors: M. Acin-Albiac, P. Filannino, R. Coda, Carlo G. Rizzello, M. Gobbetti, R. Di Cagno

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Demand for smart management of large amounts of agro-food by-products has become an area of major environmental and economic importance worldwide. Brewers' spent grain (BSG), the most abundant by-product generated in the beer-brewing process, represents an example of valuable raw material and source of health-promoting compounds. To the date, the valorization of BSG as a food ingredient has been limited due to poor technological and sensory properties. Tailored bioprocessing through lactic acid bacteria (LAB) fermentation is a versatile and sustainable means for the exploitation of food industry by-products. Indigestible carbohydrates (e.g., hemicelluloses and celluloses), high phenolic content, and mostly lignin make of BSG a hostile environment for microbial survival. Hence, the selection of tailored starters is required for successful fermentation. Our study investigated the metabolic strategies of Leuconostoc pseudomesenteroides and Lactobacillus plantarum strains to exploit BSG as a food ingredient. Two distinctive BSG samples from different breweries (Italian IT- and Finish FL-BSG) were microbially and chemically characterized. Growth kinetics, organic acid profiles, and the evolution of phenolic profiles during the fermentation in two BSG model media were determined. The results were further complemented with gene expression targeting genes involved in the degradation cellulose, hemicelluloses building blocks, and the metabolism of anti-nutritional factors. Overall, the results were LAB genus dependent showing distinctive metabolic capabilities. Leuc. pseudomesenteroides DSM 20193 may degrade BSG xylans while sucrose metabolism could be furtherly exploited for extracellular polymeric substances (EPS) production to enhance BSG pro-technological properties. Although L. plantarum strains may follow the same metabolic strategies during BSG fermentation, the mode of action to pursue such strategies was strain-dependent. L. plantarum PU1 showed a great preference for β-galactans compared to strain WCFS1, while the preference for arabinose occurred at different metabolic phases. Phenolic compounds profiling highlighted a novel metabolic route for lignin metabolism. These findings will allow an improvement of understanding of how lactic acid bacteria transform BSG into economically valuable food ingredients.

Keywords: brewery by-product valorization, metabolism of plant phenolics, metabolism of lactic acid bacteria, gene expression

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Authors: Shohei Matsuo, Saki Mikai, Hiroshi Morita

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Objectives: Shochu is a popular Japanese distilled spirits. In the production of shochu, the filamentous fungus Aspergillus kawachii has traditionally been used. A. kawachii produces two types of starch hydrolytic enzymes, α-amylase (enzymatic liquefaction) and glucoamylase (enzymatic saccharification). Liquid culture system is a relatively easy microorganism to ferment with relatively low cost of production compared for solid culture. In liquid culture system, acid-unstable α-amylase (α-A) was produced abundantly, but, acid-stable α-amylase (Aα-A) was not produced. Since there is high enzyme productivity, most in shochu brewing have been adopted by a solid culture method. In this study, therefore, we investigated production of Aα-A in liquid culture system. Materials and methods: Microorganism Aspergillus kawachii NBRC 4308 was used. The mold was cultured at 30 °C for 7~14 d to allow formation of conidiospores on slant agar medium. Liquid Culture System: A. kawachii was cultured in a 100 ml of following altered SLS medium: 1.0 g of rice flour, 0.1 g of K2HPO4, 0.1 g of KCl, 0.6 g of tryptone, 0.05 g of MgSO4・7H2O, 0.001 g of FeSO4・7H2O, 0.0003 g of ZnSO4・7H2O, 0.021 g of CaCl2, 0.33 of citric acid (pH 3.0). The pH of the medium was adjusted to the designated value with 10 % HCl solution. The cultivation was shaking at 30 °C and 200 rpm for 72 h. It was filtered to obtain a crude enzyme solution. Aα-A assay: The crude enzyme solution was analyzed. An acid-stable α-amylase activity was carried out using an α-amylase assay kit (Kikkoman Corporation, Noda, Japan). It was conducted after adding 9 ml of 100 mM acetate buffer (pH 3.0) to 1 ml of the culture product supernatant and acid treatment at 37°C for 1 h. One unit of a-amylase activity was defined as the amount of enzyme that yielded 1 mmol of 2-chloro-4-nitrophenyl 6-azide-6-deoxy-b-maltopentaoside (CNP) per minute. Results and Conclusion: We experimented with co-culture of A. kawachii and lactobacillus in order to get control of pH in altered SLS medium. However, high production of acid-stable α-amylase was not obtained. We experimented with yoghurt or milk made an addition to liquid culture. The result indicated that high production of acid-stable α-amylase (964 U/g-substrate) was obtained when milk made an addition to liquid culture. Phosphate concentration in the liquid medium was a major cause of increased acid-stable α-amylase activity. In liquid culture, acid-stable α-amylase activity was enhanced by milk, but Fats and oils in the milk were oxidized. In addition, Tryptone is not approved as a food additive in Japan. Thus, alter SLS medium added to skim milk excepting for the fats and oils in the milk instead of tryptone. The result indicated that high production of acid-stable α-amylase was obtained with the same effect as milk.

Keywords: acid-stable α-amylase, liquid culture, milk, shochu

Procedia PDF Downloads 284

Authors: Noah Emil Glisik

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Alcohol use disorder (AUD) presents significant health challenges worldwide and is particularly concerning in Slovenia, where high alcohol consumption contributes to elevated rates of comorbidities, including depression and suicide. This review examines emerging evidence linking gut microbiome dysbiosis to AUD, exploring whether gut microbiome alterations merely result from alcohol use or actively contribute to the persistence of addiction. Additionally, it discusses how microbial changes may influence psychological symptoms, including anxiety and depressive states, which are closely associated with suicidality in this population. To address gaps in existing research, a systematic literature search was conducted through PubMed, Web of Science, and ScienceDirect. Inclusion criteria focused on studies examining gut microbiome changes in AUD, particularly those assessing gut-brain axis interactions and microbial species impacting inflammation and neurotransmitter pathways. Studies were excluded if they lacked peer review or did not specifically assess microbiome effects on mental health outcomes. A qualitative literature review approach was applied, synthesizing findings into key themes on microbial changes, neuroinflammatory pathways, and treatment implications. Data were organized into tables to provide a clear comparison of microbiota alterations across studies, highlighting specific bacterial species and their potential effects on AUD. This review emphasizes patterns in AUD patients, where reductions in anti-inflammatory species, such as Faecalibacterium prausnitzii and Roseburia intestinalis, coincide with increases in pro-inflammatory bacteria like Enterococcus faecalisand Lactobacillus rhamnosus. These shifts contribute to increased gut permeability and systemic inflammation, potentially influencing the kynurenine pathway, which is linked to depressive symptoms and elevated alcohol cravings. Furthermore, the review explores the potential of probiotic therapies targeting these microbial imbalances as adjunctive treatments for AUD, particularly those focusing on strains that support anti-inflammatory pathways and gut barrier integrity. Restoring microbial homeostasis through probiotics or fecal microbiota transplantation may not only reduce inflammation but also alleviate mental health symptoms associated with addiction, including suicidality. The findings underscore the need for further clinical trials assessing microbiome-targeted therapies as innovative, multifaceted approaches to AUD treatment in Slovenia and beyond.

Keywords: alcohol use disorder, gut-brain axis, microbiome dysbiosis, probiotic therapy.

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Authors: M. Naimi, M. B. Khaled

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The aim of the present work was to test in vitro inhibition of food pathogens and spoilage bacteria by crude bacteriocins from autochthonous lactic acid bacteria. Thirty autochthonous lactic acid bacteria isolated previously, belonging to the genera: Lactobacillus, Carnobacterium, Lactococcus, Vagococcus, Streptococcus, and Pediococcus, have been screened by an agar spot test and a well diffusion assay against Gram-positive and Gram-negative harmful bacteria: Bacillus cereus, Bacillus subtilis ATCC 6633, Escherichia coli ATCC 8739, Salmonella typhimurium ATCC 14028, Staphylococcus aureus ATCC 6538, and Pseudomonas aeruginosa under conditions means to reduce lactic acid and hydrogen peroxide effect to select bacteria with high bacteriocinogenic potential. Furthermore, crude bacteriocins semiquantification and heat sensitivity to different temperatures (80, 95, 110°C, and 121°C) were performed. Another exploratory test concerning the response of St. aureus ATCC 6538 to the presence of crude bacteriocins was realized. It has been observed by the agar spot test that fifteen candidates were active toward Gram-positive targets strains. The secondary screening demonstrated an antagonistic activity oriented only against St. aureus ATCC 6538, leading to the selection of five isolates: Lm14, Lm21, Lm23, Lm24, and Lm25 with a larger inhibition zone compared to the others. The ANOVA statistical analysis reveals a small variation of repeatability: Lm21: 0.56%, Lm23: 0%, Lm25: 1.67%, Lm14: 1.88%, Lm24: 2.14%. Conversely, slight variation was reported in terms of inhibition diameters: 9.58± 0.40, 9.83± 0.46, and 10.16± 0.24 8.5 ± 0.40 10 mm for, Lm21, Lm23, Lm25, Lm14and Lm24, indicating that the observed potential showed a heterogeneous distribution (BMS = 0.383, WMS = 0.117). The repeatability coefficient calculated displayed 7.35%. As for the bacteriocins semiquantification, the five samples exhibited production amounts about 4.16 for Lm21, Lm23, Lm25 and 2.08 AU/ml for Lm14, Lm24. Concerning the sensitivity the crude bacteriocins were fully insensitive to heat inactivation, until 121°C, they preserved the same inhibition diameter. As to, kinetic of growth , the µmax showed reductions in pathogens load for Lm21, Lm23, Lm25, Lm14, Lm24 of about 42.92%, 84.12%, 88.55%, 54.95%, 29.97% in the second trails. Inversely, this pathogen growth after five hours displayed differences of 79.45%, 12.64%, 11.82%, 87.88%, 85.66% in the second trails, compared to the control. This study showed potential inhibition to the growth of this food pathogen, suggesting the possibility to improve the hygienic food quality.

Keywords: exploratory test, lactic acid bacteria, crude bacteriocins, spoilage, pathogens

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Authors: N. Selvi Gunel, L. M. Oktay, H. Memmedov, B. Durmaz, H. Kalkan Yildirim, E. Yildirim Sozmen

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Object: Propolis which consists of compounds that are accepted as antioxidant, antimicrobial, antiseptic, antibacterial, anti-inflammatory, anti-mutagenic, immune-modulator and cytotoxic, is frequently used in current therapeutic applications. However, some of them result in allergic side effects, causing consumption to be restricted. Previously our group has succeeded in producing a new biotechnological product which was less allergenic. In this study, we purpose to optimize production conditions of this biologically-transformed propolis and determine the cytotoxic effects of obtained new products on colon cancer cell line (HCT-116). Method: Firstly, solid propolis samples were dissolved in water after weighing, grinding and sizing (sieve-35mesh) and applied 40 kHz/10 min ultrasonication. Samples were prepared according to inoculation with Lactobacillus plantarum in two different proportions (2.5% and 3.5%). Chromatographic analyzes of propolis were performed by UPLC-MS/MS (Waters, Milford, MA) system. Results were analysed by UPLC-MS/MS system MassLynx™ 4.1 software. HCT-116 cells were treated with propolis examples at 25-1000 µg/ml concentrations and cytotoxicity were measured by using WST-8 assay at 24, 48, and 72 hours. Samples with biological transformation were compared with the non-transformed control group samples. Our experiment groups were formed as follows: untreated (group 1), propolis dissolved in water ultrasonicated at 40 kHz/10 min (group 2), propolis dissolved in water ultrasonicated at 40 kHz/10 min and inoculated 2.5% L. plantarum L1 strain (group 3), propolis dissolved in water ultrasonicated at 40 kHz/10 min and inoculated 3.5% L. plantarum L3 strain (group 4). Obtained data were calculated with Graphpad Software V5 and analyzed by two-way ANOVA test followed by Bonferroni test. Result: As a result of our study, the cytotoxic effect of propolis samples on HCT-116 cells was evaluated. There was a 7.21 fold increase in group 3 compared to group 2 in the concentration of 1000 µg/ml, and it was a 6.66 fold increase in group 3 compared to group 1 at the end of 24 hours. At the end of 48 hours, in the concentration of 500 µg/ml, it was determined 4.7 fold increase in group 4 compared to group 3. At the same time, in the concentration of 750 µg/ml it was determined 2.01 fold increase in group 4 compared to group 3 and in the same concentration, it was determined 3.1 fold increase in group 4 compared to group 2. Also, at the 72 hours, in the concentration of 750 µg/ml, it was determined 2.42 fold increase in group 3 according to group 2 and in the same time, in the concentration of 1000 µg/ml, it was determined 2.13 fold increase in group 4 according to group 2. According to cytotoxicity results, the group which were ultrasonicated at 40 kHz/10min and inoculated 3.5% L. plantarum L3-strain had a higher cytotoxic effect. Conclusion: It is known that bioavailability of propolis is halved in six months. The data obtained from our results indicated that biologically-transformed propolis had more cytotoxic effect than non-transformed group on colon cancer cells. Consequently, we suggested that L. plantarum-transformation provides both reduction of allergenicity and extension of bioavailability period by enhancing healthful polyphenols.

Keywords: bio-transformation, propolis, colon cancer, cytotoxicity

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Authors: A. M. Isakhanyan, F. N. Tkhruni, N. N. Yakimovich, Z. I. Kuvaeva, T. V. Khachatryan

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Introduction: At present, the simbiotics based on different genera and species of lactic acid bacteria (LAB) and yeasts are used. One of the basic properties of probiotics is presence of antimicrobial activity and therefore selection of LAB and yeast strains for their co-cultivation with the aim of increasing of the activity is topical. Since probiotic yeast and bacteria have different mechanisms of action, natural synergies between species, higher viability and increasing of antimicrobial activity might be expected from mixing both types of probiotics. Endemic strains of LAB Enterococcus faecium БТK-64, Lactobaccilus plantarum БТK-66, Pediococcus pentosus БТK-28, Lactobacillus rhamnosus БТK-109 and Kluyveromyces lactis БТX-412, Saccharomycopsis sp. БТX- 151 strains of yeast, with probiotic properties and hight antimicrobial activity, were selected. Strains are deposited in "Microbial Depository Center" (MDC) SPC "Armbiotechnology". Methods: LAB and yeast strains were isolated from different dairy products from rural households of Armenia. The genotyping by 16S rRNA sequencing for LAB and 26S RNA sequencing for yeast were used. Combined cultivation of LAB and yeast strains was carried out in the nutrient media on the basis of milk whey, in anaerobic conditions (without shaker, in a thermostat at 37oC, 48 hours). The complex preparations were obtained by purification of cell free culture broth (CFC) broth by the combination of ion-exchange chromatography and gel filtration methods. The spot-on-lawn method was applied for determination of antimicrobial activity and expressed in arbitrary units (AU/ml). Results. The obtained data showed that at the combined growth of bacteria and yeasts, the cultivation conditions (medium composition, time of growth, genera of LAB and yeasts) affected the display of antimicrobial activity. Purification of CFC broth allowed obtaining partially purified antimicrobial complex preparation which contains metabiotics from both bacteria and yeast. The complex preparation inhibited the growth of pathogenic and conditionally pathogenic bacteria, isolated from various internal organs from diseased animals and poultry with greater efficiency than the preparations derived individually alone from yeast and LAB strains. Discussion. Thus, our data shown perspectives of creation of a new class of antimicrobial preparations on the basis of combined cultivation of endemic strains of LAB and yeast. Obtained results suggest the prospect of use of the partially purified complex preparations instead antibiotics in the agriculture and for food safety. Acknowledgments: This work was supported by the RA MES State Committee of Science and Belarus National Foundation for Basic Research in the frames of the joint Armenian - Belarusian joint research project 13РБ-064.

Keywords: co-cultivation, antimicrobial activity, biosafety, metabiotics, lactic acid bacteria, yeast

Procedia PDF Downloads 339

Authors: Fatima Bouazza, Khadija Khedid, Lamiae Amallah, Aziz Mouhaddach, Basma Boukour, Jihane Ennadir, Rachida Hassikou

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In order to combine milk and its derived products conservation and flavoring, Moroccans often used aromatic and medicinal plants. These plant extracts are endowed with several nutritive and therapeutic properties. This study constitutes a first national assessment of physico-chemical quality of sheep’s milk from moroccan Sardi breed and the evaluation of the antibacterial effect of three medicinal plants extracts: Aloe barbadensis Miller, Thymus satureioides and Mentha pulegium on flora isolated from this sheep's milk. 100 milk samples were collected in four regions of Morocco. The bacteria isolated were identified by classical and molecular methods (16S rRNA sequencing) and tested, according to the disk method, for their sensitivity to several antibiotics. The physico-chemical analyzes of sheep’s milk concerned the pH, titratable acidity, density, dry extract, freezing point and contents of: fat, proteins, lactose and calcium. The essential oils (EOs) of T. satureioides and M .pulegium were extracted by hydrodistillation and analyzed by GC / MS, while the Aloe vera leaf pulp was analyzed by the methods of Harborne and HPLC. A total number of 125 bacteria have been identified. Significant resistance to chemical antibiotics has been noted in LABs. The average temperature value of milk is around 57.15 °C, the pH is 6.56, the titratable acidity is around 3.4 ° D, the density is 1.035g / cm³ , the total dry extract is around 169.5g / l, the ash (9.8g / l), the freezing point (- 0.556 °C) while the average fat content is 67.85g / l . The samples richest in fat belong to the region of Settat, cradle of the Sardi breed, with a maximum average value of 74.4g / l. The average protein is 56g / l, lactose (39.92g / l), and calcium (1.855g / l). Analysis of the major components of EOs revealed the dominance of borneol in the case of T. satureioides and of pulegone in M. pulegium. Aloe vera gel contains alkaloids, flavonoids, catechic tannins, saponins and 1.60 µg / ml of aloin. The plant extracts have a bactericidal effect on E. coli, Klebsiellaoxytoca and Staphylococci and bacteriostatic effect on LABs of technological interest (Lactobacillus). As a result of this study, it is believed that the consumption of sardi sheep’s milk would be of nutritional benefit. Its richness in fat and proteins predisposes it for biotechnological development in the manufacture of cheese and yogurt. Also, the use of aromatic and medicinal plants, as natural additives would be of great benefit to flavor and maintain its quality.

Keywords: sheep’s milk, lactic flora, antimicrobial power, aloe barbadensis miller, thymus satureioides, mentha pulegium

Procedia PDF Downloads 124

Authors: Vanessa G. P. Severino, Eliangela Cristina Candida Costa, Nubia Alves Mariano Teixeira Pires Gomides, Lucilia Kato, Afif Felix Monteiro, Maria Anita Lemos Vasconcelos Ambrosio, Carlos Henrique Gomes Martins

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One of the biomes present in Brazil is known as Cerrado, which is a vast tropical savanna ecoregion, particularly in the states of Goiás, Mato Grosso do Sul, Mato Grosso, Tocantins and Minas Gerais. Many species of plants are characterized as endemic and they have therapeutic value for a large part of the population, especially to the rural communities. Given that, the southeastern region of the state of Goiás contains about 21 rural communities, which present a form of organization based on the use of natural resources available. One of these rural communities is named of Coqueiros, where the knowledge about the medicinal plants was very important to this research. Thus, this study focuses on the ethnobotanical survey of this community on the use of Kielmeyera coriacea to treat diseases. From the 37 members interviewed, 76% indicated this species for the treatment of intestinal infection, leukemia, anemia, gastritis, gum pain, toothache, cavity, arthritis, arthrosis, healing, vermifuge, rheumatism, antibiotic, skin problems, mycoses and all kinds of infections. The medicinal properties attributed during the interviews were framed in the body system (disease categories), adapted from ICD 10; thus, 20 indications of use were obtained, among five body systems. Therefore, the root of this species was select to chemical and biological (antioxidant and antimicrobial) studies. From the liquid-liquid extraction of ethanolic extract of root (EER), the hexane (FH), ethyl acetate (FAE), and hydro alcoholic (FHA) fractions were obtained. The chemical profile study of these fractions was performed by LC-MS, identifying major compounds such as δ-tocotrienol, prenylated acylphoroglucinol, 2-hydroxy-1-methoxyxanthone and quercitrin. EER, FH, FAE and FHA were submitted to biological tests. FHA presented the best antioxidant action (EC50 201.53 μg mL-1). EER inhibited the bacterial growth of Streptococcus pyogenes and Pseudomonas aeruginosa, microorganisms associated with rheumatism, at Minimum Inhibitory Concentration (MIC) of 6.25 μg mL-1. In addition, the FH-10 subfraction, obtained from FH fractionation, presented MIC of 1.56 μg mL-1 against S. pneumoniae; EER also inhibited the fungus Candida glabrata (MIC 7.81 μg mL- 1). The FAE-4.7.3 fraction, from the fractionation of FAE, presented MIC of 200 μg mL-1 against Lactobacillus casei, which is one of the causes of caries and oral infections. By the correlation of the chemical and biological data, it is possible to note that the FAE-4.7.3 and FH-10 are constituted 4-hydroxy-2,3-methylenedioxy xanthone, 3-hydroxy-1,2-dimethoxy xanthone, lupeol, prenylated acylphoroglucinol and quercitrin, which could be associated with the biological potential found. Therefore, this study provides an important basis for further investigations regarding the compounds present in the active fractions of K. coriacea, which will permit the establishment of a correlation between ethnobotanical survey and bioactivity.

Keywords: biological activity, ethnobotanical survey, Kielmeyera coriacea Mart., LC-MS profile

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Authors: António Inês, Ana Guimarães, José Maria, Vânia Laranjo, Armando Venâncio, Luís Abrunhosa

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Lactic acid bacteria (LAB) play a key role in the biopreservation of a wide range of fermented food products, such as yogurt, cheese, fermented milks, meat, fish, vegetables (sauerkraut, olives and pickles), certain beer brands, wines and silage, allowing their safe consumption, which gave to these bacteria a GRAS (Generally Recognised as Safe) status. Besides that, the use of LAB in food and feed is a promising strategy to reduce the exposure to dietary mycotoxins, improving their shelf life and reducing health risks, given the unique mycotoxin decontaminating characteristic of some LAB. Mycotoxins present carcinogenic, mutagenic, teratogenic, neurotoxic and immunosuppressive effects over animals and Humans, being the most important ochratoxin A (OTA), aflatoxins (AFB1), trichothecenes, zearalenone (ZEA), fumonisin (FUM) and patulin. In a previous work of our group it was observed OTA biodegradation by some strains of Pediococcus parvulus isolated from Douro wines. So, the aim of this study was to enlarge the screening of the biodetoxification over more mycotoxins besides OTA, including AFB1, and ZEA. This ability was checked in a collection of LAB isolated from vegetable (wine, olives, fruits and silage) and animal (milk and dairy products, sausages) sources. All LAB strains were characterized phenotypically (Gram, catalase) and genotypically. Molecular characterisation of all LAB strains was performed using genomic fingerprinting by MSP-PCR with (GTG)5 and csM13 primers. The identification of the isolates was confirmed by 16S rDNA sequencing. To study the ability of LAB strains to degrade OTA, AFB1 and ZEA, a MRS broth medium was supplemented with 2.0 μg/mL of each mycotoxin. For each strain, 2 mL of MRS supplemented with the mycotoxins was inoculated in triplicate with 109 CFU/mL. The culture media and bacterial cells were extracted by the addition of an equal volume of acetonitrile/methanol/acetic acid (78:20:2 v/v/v) to the culture tubes. A 2 mL sample was then collected and filtered into a clean 2 mL vial using PP filters with 0.45 μm pores. The samples were preserved at 4 °C until HPLC analysis. Among LAB tested, 10 strains isolated from milk were able to eliminate AFB1, belonging to Lactobacillus casei (7), Lb. paracasei (1), Lb. plantarum (1) and 1 to Leuconostoc mesenteroides. Two strains of Enterococcus faecium and one of Ec. faecalis from sausage eliminated ZEA. Concerning to strains of vegetal origin, one Lb. plantarum isolated from elderberry fruit, one Lb. buchnerii and one Lb. parafarraginis both isolated from silage eliminated ZEA. Other 2 strains of Lb. plantarum from silage were able to degrade both ZEA and OTA, and 1 Lb. buchnerii showed activity over AFB1. These enzymatic activities were also verified genotypically through specific gene PCR and posteriorly confirmed by sequencing analysis. In conclusion, due the ability of some strains of LAB isolated from different sources to eliminate OTA, AFB1 and ZEA one can recognize their potential biotechnological application to reduce the health hazards associated with these mycotoxins. They may be suitable as silage inoculants or as feed additives or even in food industry.

Keywords: bio-detoxification, lactic acid bacteria, mycotoxins, food and feed

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Authors: Anuj Tyagi, Balwinder Singh, Naveen Kumar B. T., Niraj K. Singh

Abstract:

Characterization of diverse microbial communities in specific environment plays a crucial role in the better understanding of their functional relationship with the ecosystem. It is now well established that gut microbiome of fish is not the simple replication of microbiota of surrounding local habitat, and extensive species, dietary, physiological and metabolic variations in fishes may have a significant impact on its composition. Moreover, overuse of antibiotics in human, veterinary and aquaculture medicine has led to rapid emergence and propagation of antibiotic resistance genes (ARGs) in the aquatic environment. Microbial communities harboring specific ARGs not only get a preferential edge during selective antibiotic exposure but also possess the significant risk of ARGs transfer to other non-resistance bacteria within the confined environments. This phenomenon may lead to the emergence of habitat-specific microbial resistomes and subsequent emergence of virulent antibiotic-resistant pathogens with severe fish and consumer health consequences. In this study, gut microbiota of freshwater carp (Labeo rohita) was investigated by shotgun metagenomics to understand its taxonomic composition and functional capabilities. Metagenomic DNA, extracted from the fish gut, was subjected to sequencing on Illumina NextSeq to generate paired-end (PE) 2 x 150 bp sequencing reads. After the QC of raw sequencing data by Trimmomatic, taxonomic analysis by Kraken2 taxonomic sequence classification system revealed the presence of 36 phyla, 326 families and 985 genera in the fish gut microbiome. At phylum level, Proteobacteria accounted for more than three-fourths of total bacterial populations followed by Actinobacteria (14%) and Cyanobacteria (3%). Commonly used probiotic bacteria (Bacillus, Lactobacillus, Streptococcus, and Lactococcus) were found to be very less prevalent in fish gut. After sequencing data assembly by MEGAHIT v1.1.2 assembler and PROKKA automated analysis pipeline, pathway analysis revealed the presence of 1,608 Metacyc pathways in the fish gut microbiome. Biosynthesis pathways were found to be the most dominant (51%) followed by degradation (39%), energy-metabolism (4%) and fermentation (2%). Almost one-third (33%) of biosynthesis pathways were involved in the synthesis of secondary metabolites. Metabolic pathways for the biosynthesis of 35 antibiotic types were also present, and these accounted for 5% of overall metabolic pathways in the fish gut microbiome. Fifty-one different types of antibiotic resistance genes (ARGs) belonging to 15 antimicrobial resistance (AMR) gene families and conferring resistance against 24 antibiotic types were detected in fish gut. More than 90% ARGs in fish gut microbiome were against beta-lactams (penicillins, cephalosporins, penems, and monobactams). Resistance against tetracycline, macrolides, fluoroquinolones, and phenicols ranged from 0.7% to 1.3%. Some of the ARGs for multi-drug resistance were also found to be located on sequences of plasmid origin. The presence of pathogenic bacteria and ARGs on plasmid sequences suggested the potential risk due to horizontal gene transfer in the confined gut environment.

Keywords: antibiotic resistance, fish gut, metabolic pathways, microbial diversity

Procedia PDF Downloads 144

Authors: Sandhya Parameswaran, Prajakta Dhuri

Abstract:

Despite great improvements in health care, the world oral health report states that dental problems still persist, particularly among underprivileged groups in both developing and developed countries. Dental caries and periodontal diseases are identified as the most important oral health problems globally. Acidic foods and beverages can affect natural teeth, and chronic exposure often leads to the development of dental erosion, abrasion, and decay. In recent years, there has been an increased interest toward essential oils. These are secondary metabolites and possess antibacterial, antifungal and antioxidant properties. Essential oils are volatile and chemically unstable in the presence of air, light, moisture and high temperature. Hence many novel methods like a liposomal encapsulation of oils have been introduced to enhance the stability and bioavailability. This research paper focuses on two essential oils, clove and cinnamon oil. Clove oil was obtained from Syzygium aromaticum Linn using clavengers apparatus. It contains eugenol and β caryophyllene. Cinnamon oil, from the barks of Cinnamomum cassia, contains cinnamaldehyde, The objective of the current research was to develop a liposomal carrier system containing clove and cinnamon oil and study their synergistic activity against dental pathogens when formulated as a gel. Methodology: The essential oil were first tested for their antimicrobial activity against dental pathogens, Lactobacillus acidophillus (MTCC No. 10307, MRS broth) and Streptococcus Mutans (MTCC No .890, Brain Heart Infusion agar). The oils were analysed by UV spectroscopy for eugenol and cinnamaldehyde content. Standard eugenol was linear between 5ppm to 25ppm at 282nm and standard cinnamaldehde from 1ppm to 5pmm at 284nm. The concentration of eugenol in clove oil was found to be 62.65 % w/w, and that of cinnamaldehyde was found to be 5.15%s w/w. The oils were then formulated into liposomes. Liposomes were prepared by thin film hydration method using Phospholipid, Cholesterol, and other oils dissolved in a chloroform methanol (3:1) mixture. The organic solvent was evaporated in a rotary evaporator above lipid transition temperature. The film was hydrated with phosphate buffer (pH 5.5).The various batches of liposomes were characterized and compared for their size, loading rate, encapsulation efficiency and morphology. The prepared liposomes when evaluated for entrapment efficiency showed 65% entrapment for clove and 85% for cinnamon oil. They were also tested for their antimicrobial activity against dental pathogens and their synergistic activity studied. Based on the activity and the entrapment efficiency the amount of liposomes required to prepare 1gm of the gel was calculated. The gel was prepared using a simple ointment base and contained 0.56% of cinnamon and clove liposomes. A simultaneous method of analysis for eugenol and cinnamaldehyde.was then developed using HPLC. The prepared gels were then studied for their stability as per ICH guidelines. Conclusion: It was found that liposomes exhibited spherical shaped vesicles and protected the essential oil from degradation. Liposomes, therefore, constitute a suitable system for encapsulation of volatile, unstable essential oil constituents.

Keywords: cinnamon oil, clove oil, dental caries, liposomes

Procedia PDF Downloads 194