Positive and negative aspects of bacteriophages and their immense role in the food chain.

Bacteriophages infect and replicate inside a bacterial host as well as serve as natural bio-control agents. Phages were once viewed as nuisances that caused fermentation failures with cheese-making and other industrial processes, which lead to economic losses, but phages are now increasingly being observed as being promising antimicrobials that can fight against spoilage and pathogenic bacteria. Pathogen-free meals that fulfil industry requirements without synthetic additives are always in demand in the food sector. This study introduces the readers to the history, sources, and biology of bacteriophages, which include their host ranges, absorption mechanisms, lytic profiles, lysogenic profiles, and the influence of external factors on the growth of phages. Phages and their derivatives have emerged as antimicrobial agents, biodetectors, and biofilm controllers, which have been comprehensively discussed in addition to their potential applications in the food and gastrointestinal tract, and they are a feasible and safe option for preventing, treating, and/or eradicating contaminants in various foods and food processing environments. Furthermore, phages and phage-derived lytic proteins can be considered potential antimicrobials in the traditional farm-to-fork context, which include phage-based mixtures and commercially available phage products. This paper concludes with some potential safety concerns that need to be addressed to enable bacteriophage use efficiently.

In vitro safety assessment of electrohydrodynamically encapsulated Lactiplantibacillus plantarum CRD7 and Lacticaseibacillus rhamnosus CRD11 for probiotics use.

The current study aimed to validate the safety of electrohydrodynamically encapsulated Lactiplantibacillus plantarum CRD7 and Lacticaseibacillus rhamnosus CRD11 in accordance with guidelines of FAO/WHO and ICMR/DBT. In vitro assays such as mucin degradation, hemolysis of blood cells, antimicrobial susceptibility pattern, possession of virulence factors, biogenic amine, and ammonia production were assessed. In results, the cross-streak and co-culture techniques revealed that CRD7 and CRD11 were compatible in vitro. Upon visual inspection through scanning electron and fluorescence microscopy, the integrity of bacterial cell membrane was confirmed even after the encapsulation process. CRD7 and CRD11 were non-hemolytic and showed negative responses to gelatinase, urease, and DNase activities. Non-mucinolytic activity of CRD7 and CRD11 was verified by measuring cell growth rate (p < 0.05) in different modified media followed by SDS-PAGE. High-performance liquid chromatography analysis revealed that both the strains did not produce biogenic amines (putrescine, cadaverine, histamine, and tyramine). Neither of the Lactobacillus strains produced ammonia after growing in BHI broth for 5 days at 37 °C. L-lactate production was highest (p < 0.05) in CRD11 (8.83 g/L), followed by CRD7 (8.16 g/L), whereas the lowest (p < 0.05) D-lactate was registered for encapsulated CRD11 (0.33 g/L) and CRD7 (0.49 g/L). The antibiogram profile determined through minimum inhibitory concentration showed that CRD7 and CRD11 were sensitive to key antibiotics suggested by EFSA except for gentamycin and kanamycin. PCR data on virulence genes demonstrated that both strains were safe for probiotic use. Moreover, CRD7 and CRD11 strains caused insignificant (p > 0.05) changes in the cell viability of Caco-2 cells as estimated by MTT (98.94-99.50%) and NR uptake (95.42-97.03%) assays and showed sensitivity to human serum. According to the results of these evaluated attributes, it is concluded that L. plantarum CRD7 and L. rhamnosus CRD11 are safe, non-toxic to human epithelial cells, and thus may be potentially suitable for various food/feed applications.

Antibacterial activity and mechanism of essential oils in combination with medium-chain fatty acids against predominant bovine mastitis pathogens.

Bovine mastitis has become a significant economic importance for the dairy industry. Concerns regarding poor milk quality and emergence of bacterial resistance have necessitated to develop an alternative therapeutic approach to antibiotics for the treatment of mastitis. Saturated medium-chain fatty acids (MCFAs) and essential oils (EOs) are known natural antimicrobials, but their combined effect has not been investigated extensively. The objective of the present investigation was to examine the bactericidal effect of various combined treatments of eight EOs and three saturated MCFAs to inactivate predominant mastitis pathogens, including Staphylococcus aureus ATCC 29213; Escherichia coli ATCC 25922; Klebsiella pneumoniae ATCC 27736 and Streptococcus agalactiae ATCC 27956. The minimum inhibitory concentration (MIC) values confirmed that all the tested pathogens were variably susceptible to both EOs and saturated MCFAs. Among essential oils, carvacrol (CAR), trans-cinnamaldehyde (TC) and thymol (TM) showed the highest inhibitory activity at concentration 0·38-1·32 mg/mL. Carvacrol exhibited effective additive antibacterial activity in combined treatment with octanoic acid (OA) in terms of its fractional inhibitory index (0·63-0·88) and time-kill effect in reducing about 6 log CFU/mL bacterial cells in less than 5 min. The effort was also made to elucidate the mechanism of antibacterial action of CAR and OA against selected mastitis pathogens by observing changes in cell microstructure, permeability and integrity of cell membrane and their membrane potential. After adding CAR and OA at MIC level, there were obvious changes in cell morphology, leakage of small electrolytes and macromolecules at the initial few hours of treatment i.e. within 1-2 h were observed. Our results indicated that CAR and OA could be evaluated as alternatives or adjuncts to antibiotics as intramammary infusion or topical application to treat bovine mastitis, significantly improving the microbiological safety of milk.

Efficacy and mechanism of carvacrol with octanoic acid against mastitis causing multi-drug-resistant pathogens.

In the present investigation, we determined the in vitro antimicrobial activity of eight essential oils (EOs) and three medium-chain fatty acids (MCFAs) alone and in combination against Staphylococcus aureus ATCC 700698, Klebsiella pneumoniae ATCC 700603, and E. coli FcW5. The interactions between EOs and MCFAs were determined by fractional inhibitory concentration indices. Moreover, mode of action of selected bioactive components was studied by changes in bacterial surface charge, morphology, and membrane integrity assays. Among EOs, carvacrol (CAR), trans-cinnamaldehyde (TC), and thymol (TM) showed strong antimicrobial activity. In combination study, CAR+OA (octanoic acid), CAR+DA (decanoic acid), and TM+OA were observed as the most significant (P≤0.05) which were also confirmed through time-kill plots. Based on these results, CAR+OA were found to be most efficacious in terms of killing time (P≤0.05). Changes in the surface charge, morphology, and membrane integrity upon the combined treatment of CAR+OA were also observed, which ultimately leads to cell death. Results suggest that CAR+OA when used in combination offer a significant (P≤0.05) additive antimicrobial activity against the selected pathogenic bacteria. Therefore, these natural bioactive molecules could be interesting alternatives to conventional therapy for the control of mastitis caused by multi-drug-resistant pathogens in bovine animals to ensure the milk safety.

Aflatoxin M1 Detoxification Ability of Probiotic Lactobacilli of Indian Origin in In vitro Digestion Model.

Aflatoxin M1 (AFM1) is known to be a potent carcinogen and continues to pose a public health concern through the consumption of contaminated dairy foods. It is anticipated that consumption of lactic acid bacteria capable of binding aflatoxins can reduce the risk of AFM1 on human health to a certain extent. Seldom reports have hinted the possibility of using lactic acid bacteria for the biological detoxification of AFM1. Hence, the present study was conducted to assess the ability of selected probiotic Lactobacillus strains for their AFM1 binding ability in PBS and to reduce its bioaccessibility in artificially contaminated skim milk using an in vitro digestion model. Eleven tested probiotic strains illustrated various degrees of AFM1 binding ability ranging from 4.13 to 64.16%. Five among the 11 probiotic strains were subsequently selected for detailed studies on the basis of highest binding potential after 24 h of incubation period. The stability of bacterial-AFM1 complex was assessed by repeated washings with AFM1 free PBS. The observation on bacterial-AFM1 complex stability showed small release of AFM1 in first and second wash (17.30 to 0.98%) where as none was detectable in the third wash. However, upon chloroform extraction, 88.57 to 92.30% of bound AFM1 was released from the bacterial cells which indicate AFM1 binding to the bacterial cell surface rather than absorption or degradation of AFM1 by bacterial cells. During the in vitro digestion test in skim milk, bioaccessibility of AFM1 was reduced to a scale of 32.61 to 52.84% in the presence of selected strains of probiotic lactobacilli. The present findings suggest that selected probiotic strains could be potentially used to mitigate the toxic effects of AFM1 in the contaminated milk and milk products and thereby enhance food safety.

A Novel Herbal Formulation versus Chlorhexidine Mouthwash in Efficacy against Oral Microflora.

AIMS AND OBJECTIVES: The aim of the study was to compare and to analyze the antimicrobial efficacy of 0.12% chlorhexidine and new formulated herbal mouthwash after using for 14 days. The objective was to signify whether the noval herbal combination could be a better alternative mouthwash to Chlorhexidine (CHX). MATERIALS AND METHODS: This is a double-blinded, random controlled research study conducted in the Department of Oral Pathology and Microbiology. A total of 200 dental students were selected randomly, comprising of two groups, 100 in each, aged between 18 and 22 years with gingival index of score II. The first group was advised to oral rinse with 0.12% chlorhexidine mouthwash and the second group with new formulated herbal mouthwash for 14 days. Saliva samples were collected on the day 0 (baseline), followed by day 7 and 14 and microbial colony count was performed. The data obtained was statistically analyzed using SPSS version 16. Student's t-test was applied for comparison of the mean microbial count between the two groups. Repeated measures analysis of variance followed by Tukey's post hoc test was applied to assess the changes from day 0 to day 7 to day 14. The statistical significance level was set at P < 0.05. RESULTS: Microbial colonies were reduced better in chlorhexidine group on the day 7 whereas, on day 14, greater reduction was observed in the herbal group in both gender groups with high statistical significance (P < 0.001). CONCLUSIONS: Herbal mouthwash formulation performed effectively well on long-term usage, could be used as an alternative mouthwash to overcome the disadvantages of chlorhexidine.

Bio-prospectus of cadmium bioadsorption by lactic acid bacteria to mitigate health and environmental impacts.

Foodstuffs and water are the key sources of cadmium biomagnifiaction. The available strategies to mitigate this problem are unproductive and expensive for practical large-scale use. Biological decontamination of metals through environmental microbes has been known since long time, whereas lactic acid bacteria (LAB) have not been extensively studied for this purpose. The LAB are known for maintaining homeostasis and suppression of pathogens in humans and animals. They also play a vital role in bioremediation of certain heavy metals. Recently in-vivo research findings strongly complement the in-vitro results in relation to decreased total body cadmium burden in animal model. This review summarizes the currently available information on impact of toxic metal (Cd) on human and animal health as well as cadmium sequestration through microbes placed broadly, whereas preeminent attention grabbed on LAB-cadmium interaction to explore their possible role in bioremediation of cadmium from foods and environment to safeguard human as well as environment health.

Impact of oral cadmium intoxication on levels of different essential trace elements and oxidative stress measures in mice: a response to dose.

The study evaluated the effect of oral intoxication of cadmium and the possible causes of oxidative stress and its preferential accumulation in different organs as well as sub-sequential effects in mice. Twenty-four Swiss albino male mice were divided into three groups viz., normal control group without cadmium chloride (CdCl2), whereas a daily dose of 0.5 and 1.2 mg of CdCl2 was orally administered for a period of a week to dose group 1 (DG-1) and dose group 2 (DG-2), respectively. A significant increase in the severity of cadmium toxicity was observed in animals as evidenced by aggravation in liver enzymes viz., serum alanine aminotransferase and aspartate transaminase, whereas lower levels of antioxidative stress markers in liver and kidney tissues of treated mice were observed as compared to normal control group. A significant depletion of calcium levels in liver tissues of DG-1 (217.36 ± 1.73 µg/g of wet tissues) and DG-2 (186.41 ± 1.56 µg/g of wet tissues) groups, along with Cd accumulation, was observed. To summarize, the current study would increase our understanding with respect to dose-dependent absorption of Cd and its toxicity led to mortality as well as adverse health effects in the body of mice. Graphical abstract ᅟ.

Efficacy of indigenous probiotic Lactobacillus strains to reduce cadmium bioaccessibility - An in vitro digestion model.

The toxic heavy metal cadmium (Cd) appears as one of the major global threats to human and animal health. Human being and aquatic life are exposed to Cd by breathing, eating, or drinking when industrial effluents released into environment. The study was aimed to identify cadmium-binding Lactobacillus strain to reduce its bioaccessibility in in vitro digestion model. In this context, forty-eight lactobacilli strains isolated and characterized from fermented dairy products and human origin were screened for their Cd biosorption potential using Flame Atomic Absorption Spectroscopy (FAAS). The present study revealed that Cd biosorption potential of 48 lactobacilli strains ranged from 1.0832 ± 0.012 to 3.562 ± 0.03 mg Cd g-1 of cells from initial 10 mg L-1 cadmium chloride (CdCl2) aqueous solution. Lactobacillus plantarum strain HD 48 demonstrated highest biosorption of 3.562 ± 0.03 mg Cd g-1 of cells. Lactobacilli-Cd complex stability indicated its strong stability as even after three washes with Milli-Q water metal desorption was nonsignificant (p < 0.05) and further studies to delineate the influence of Cd (100 mg L-1 CdCl2) on their growth. Moreover, these strains were able to reduce Cd bioaccessibility in the in vitro digestion model in the range of 24.71 to 41.62 %. Transmission electron microscopy (TEM) investigations on Cd bioadsorption also revealed its surface associated bioadsorption phenomenon. These findings depicted that probiotic strain L. plantarum HD 48 was found to be endowed with remarkable Cd biosorption ability as well as reduction in its bioaccessibility. These results suggest that probiotic strain L. plantarum HD 48 has immense potential to sequester Cd from aqueous solution which could be further explored as a potent source to diminish body Cd burden.

Extended shelf life flavoured dairy drink using dissolved carbon dioxide.

Cardamom flavoured dairy beverage prepared using standardized method was carbonated in glass bottles. Carbonation at 50 psi pressure for 30 s was recommended. The pasteurized flavoured drink, carbonated or otherwise was evaluated for sensory, chemical and microbial quality during its refrigerated storage. The uncarbonated control samples were found to be sensorily acceptable up to 14 days, while the carbonated beverage remained acceptable up to 30 days. Carbonation of drink significantly affected the pH and acidity of product without reducing its acceptability. Carbonation resulted in inhibition of microbes, the effect was pronounced on psychrotrophic count. There was a linear but marginal increase in the pH of the carbonated samples till the 17(th) day of storage; the values diminished thereafter. The carbonated samples also had significantly reduced contents of FFA and soluble nitrogen compared to that of uncarbonated control samples as storage progressed beyond 10 days and this was attributed to inhibited microbial growth.

Carbonated fermented dairy drink - effect on quality and shelf life.

Processing conditions were standardized for a carbonated sweetened fermented dairy beverage. The optimum level of carbonation for the beverage filled in 200 ml glass bottles was found to be at 50 psi pressure for 30 seconds. The beverage samples were stored under refrigerated conditions (7 °C) and evaluated at weekly intervals for their sensory, chemical and microbial quality. The uncarbonated control samples were found to keep well till 5 weeks of storage while the carbonated beverage was acceptable up to 12 weeks of storage. Carbonation did not significantly alter the pH of the beverage, while a marginal increase in titratable acidity was recorded for the carbonated samples. Carbonation was found to arrest the development of lipolysis and proteolysis in the beverage during storage. Microbiological investigations established the inhibition of yeast and mold growth due to dissolved CO2.