Lactic acid bacteria as an eco-friendly approach in plant production: Current state and prospects.

Since the late nineteenth century, the agricultural sector has experienced a tremendous increase in chemical use in response to the growing population. Consequently, the intensive and indiscriminate use of these substances caused serious damage on several levels, including threatening human health, disrupting soil microbiota, affecting wildlife ecosystems, and causing groundwater pollution. As a solution, the application of microbial-based products presents an interesting and ecological restoration tool. The use of Plant Growth-Promoting Microbes (PGPM) affected positive production, by increasing its efficiency, reducing production costs, environmental pollution, and chemical use. Among these microbial communities, lactic acid bacteria (LAB) are considered an interesting candidate to be formulated and applied as effective microbes. Indeed, these bacteria are approved by the European Food Safety Authority (EFSA) and Food and Drug Administration (FDA) as Qualified Presumption of Safety statute and Generally Recognized as Safe for various applications. To do so, this review comes as a road map for future research, which addresses the different steps included in LAB formulation as biocontrol, bioremediation, or plant growth promoting agents from the isolation process to their field application passing by the different identification methods and their various uses. The plant application methods as well as challenges limiting their use in agriculture are also discussed.

Control of Listeria monocytogenes in a fresh cheese using aromatic and medicinal plants and enterocin: a comparative study.

The aim of this study was to investigate the effectiveness of essential oils (EOs) or crude extracts (CEs) of eight aromatic and medicinal plants (AMPs) and its association with enterocin OS1 on Listeria monocytogenes and food spoilage bacteria in Moroccan fresh cheese. The cheese batches were treated with EO of Rosmarinus officinalis, Thymus vulgaris, Syzygium aromaticum, Laurus nobilis, Allium sativum, Eucalyptus globulus, or CE of Crocus sativus and Carthamus tinctorius, and/or enterocin OS1, and stored for 15 days at 8°C. The data were subjected to correlations analysis, variance analysis, and principal components analysis. Results clearly showed a positive correlation between L. monocytogenes reduction and storage time. Moreover, reduction on Listeria counts induced by Allium-EO and Eucalyptus-EO reached 2.68 and 1.93 Log CFU/g with respect to untreated samples after 15 days, respectively. Similarly, enterocin OS1 alone has significantly reduced the L. monocytogenes population with 1.46 Log CFU/g. The most promising result was the synergy observed between many AMPs and enterocin. Indeed, treatments with Eucalyptus-EO + OS1 and Crocus-CE + OS1 decreased the Listeria population to undetectable after only 2 days and throughout the storage period. These findings suggest a promising application/use of this natural combination, which preserves the safety and long-lasting conservation of fresh cheese.

Subchronic toxicity study in BALBc mice of enterocin AS-48, an anti-microbial peptide produced by Enterococcus faecalis UGRA10.

Few studies have examined the use of animal models to evaluate the in-vivo toxicity of antimicrobial peptides, but such research is essential to their safe use in foods. This study was performed to evaluate any adverse effects of enterocin AS-48, a circular bacteriocin produced by Enterococcus strains, when administered to BALB/c mice at concentrations of 50, 100, and 200 mg/kg in the diet for 90 days. Animals dosed with nisin at a dietary concentration of 200 mg/kg served as a reference treated group. There were no deaths in any of the animal groups, and the AS-48 treatment produced no abnormalities or clinical signs on body weights, food consumption, urinalysis, haematology, or blood biochemistry. Furthermore, there were no significant differences in the weights of liver, spleen, heart, kidneys, and intestines between control mice and those treated with AS-48 or nisin. The histopathological study showed moderate vacuolar degeneration in hepatocytes of some animals fed 100 or 200 mg/kg AS-48 (3/10 and 2/10 respectively). However, this anomaly was lower than in the group treated with nisin (5/10). Conclusively, no toxicologically significant changes were associated in BALB/c mice fed with 50, 100, and 200 mg/kg enterocin AS-48 for 90 days.

Environmental factors shape the community of symbionts in the hoopoe uropygial gland more than genetic factors.

Exploring processes of coevolution of microorganisms and their hosts is a new imperative for life sciences. If bacteria protect hosts against pathogens, mechanisms facilitating the intergenerational transmission of such bacteria will be strongly selected by evolution. By disentangling the diversity of bacterial strains from the uropygium of hoopoes (Upupa epops) due to genetic relatedness or to a common environment, we explored the importance of horizontal (from the environment) and vertical (from parents) acquisition of antimicrobial-producing symbionts in this species. For this purpose, we compared bacterial communities among individuals in nonmanipulated nests; we also performed a cross-fostering experiment using recently hatched nestlings before uropygial gland development and some nestlings that were reared outside hoopoe nests. The capacity of individuals to acquire microbial symbionts horizontally during their development was supported by our results, since cross-fostered nestlings share bacterial strains with foster siblings and nestlings that were not in contact with hoopoe adults or nests also developed the symbiosis. Moreover, nestlings could change some bacterial strains over the course of their stay in the nest, and adult females changed their bacterial community in different years. However, a low rate of vertical transmission was inferred, since genetic siblings reared in different nests shared more bacterial strains than they shared with unrelated nestlings raised in different nests. In conclusion, hoopoes are able to incorporate new symbionts from the environment during the development of the uropygium, which could be a selective advantage if strains with higher antimicrobial capacity are incorporated into the gland and could aid hosts in fighting against pathogenic and disease-causing microbes.

Hexavalent chromium removal by a novel Serratia proteamaculans isolated from the bank of Sebou River (Morocco).

The novel Serratia proteamaculans isolated from a chromium-contaminated site was tolerant to a concentration of 500 mg Cr(VI)/l. The optimum pH and temperature for reduction of Cr(VI) by S. proteamaculans were found to be 7.0 and 30 °C, respectively. The Cr(VI) reduction rate decreased with the increase in Cr(VI) concentration from 100 to 400 mg/l, suggesting the enzymatic chromium reduction. Resting and permeabilised cell assays provided the better evidence that chromate reduction in S. proteamaculans is enzymatic. Reduction by cell-free filtrate shows no extracellular chromate-reducing activity, revealing that this activity may be associated to membrane fraction and/or cytosolic fraction. Assays conducted with cytosolic and particulate fraction of S. proteamaculans confirmed the role of membrane-bound proteins in Cr(VI) reduction. Furthermore, chromium reduced by heat-treated cells suggests that membrane-associated chromate reductase activity of S. proteamaculans is preceded by its adsorption on the cell surface.

Combined effect of enterocin AS-48 and high hydrostatic pressure to control food-borne pathogens inoculated in low acid fermented sausages.

The single and combined effects of enterocin AS-48 and high hydrostatic pressure (HHP) on Listeria monocytogenes, Salmonellaenterica, and Staphylococcus aureus was investigated in fuet (a low acid fermented sausage) during ripening and storage at 7 degrees C or at room temperature. AS-48 (148 AU g(-1)) caused a drastic 5.5 log cfu g(-1) decrease in L. monocytogenes (P<0.001) and a significant (P<0.01) inhibition (1.79 logs) for Salmonella at the end of ripening (10 d). After pressurization (400 MPa) and storage Listeria counts remained below 5 cfu g(-1) in all fuets containing AS-48 (pressurized or not). HHP alone had no anti-Listeria effect. HHP treatment significantly reduced Salmonella counts, with lowest levels in pressurized fuets with AS-48. S. aureus showed similar growth for all treatments and storage conditions. These results indicate that AS-48 can be applied alone to control L. monocytogenes and combined with HHP treatment to control Salmonella in fuets.

Antimicrobial chemicals in hoopoe preen secretions are produced by symbiotic bacteria.

Animals frequently use metabolites produced by symbiotic bacteria as agents against pathogens and parasites. Secretions from the preen gland of birds are used for this purpose, although its chemicals apparently are produced by the birds themselves. European hoopoes Upupa epops and green woodhoopoes Phoeniculus purpureus harbour symbiotic bacteria in the uropygial gland that might be partly responsible for the chemical composition of secretions. Here we investigate the antimicrobial activity of the volatile fraction of chemicals in hoopoe preen secretions, and, by means of experimental antibiotic injections, test whether symbiotic bacteria living within the uropygial gland are responsible for their production. Hoopoes produce two different kinds of secretions that differ drastically in their chemical composition. While the malodorous dark secretions produced by nestlings included a complex mix of volatiles, these chemicals did not appear in white secretions produced by non-nesting birds. All volatiles detected showed strong antibacterial activity, and a mixture of the chemicals at the concentrations measured in nestling glands inhibited the growth of all bacterial strains assayed. We found support for the hypothesized role of bacteria in the production of such antimicrobial chemicals because experimental clearance of bacteria from glands of nestlings with antibiotics resulted in secretions without most of the volatiles detected in control individuals. Thus, the presence of symbiotic bacteria in the uropygial gland provides hoopoes with potent antimicrobials for topical use.

Control of Listeria monocytogenes in model sausages by enterocin AS-48.

In this work we describe the control of Listeria monocytogenes CECT 4032 in sausage by adding the enterocin AS-48 producer strains Enterococcus faecalis A-48-32 and Enterococcus faecium S-32-81, and also by adding a semi-purified preparation of the bacteriocin. Addition of preformed AS-48 caused a significant decrease (P<0.01) in the number of viable listeria even at the lowest bacteriocin concentration tested (112 AU/g). At a higher concentration (225 AU/g) listeria were below the detection level (1.99 log units/g) in meat at 3 days of incubation but growth of listeria was observed again after 9 days. For an AS-48 concentration of 450 AU/g, no viable listeria were detected after 6 and 9 days of incubation. When E. faecalis A-48-32 was used as inoculum at approximately 10(7) cfu/g, listeria counts decreased progressively from start of experiment, being below detection level at day 9. The best results were obtained with E. faecium S-32-81, since listeria were undetectable at 6 days of incubation. Bacteriocin concentrations in samples reached concentrations of 60 and 80 AU/g for strains A-48-32 and S-32-81, respectively. These results clearly indicate that AS-48 can be used in the control of L. monocytogenes in sausages.

Control of Staphylococcus aureus in sausages by enterocin AS-48.

Results presented here are the first contribution on the anti-staphylococcal activity of bacteriocin AS-48 in a model meat sausage system. We have examined bacteriocin application, by inoculation with the enterocin AS-48 producer strain Enterococcus faecalis A-48-32 or by adding a semi-purified bacteriocin preparation. AS-48 inhibits proliferation of Staphylococcus aureus in sausages when added at concentrations of 30 or 40µg/g, achieving a significant reduction of 2 and 5.31 log units, respectively, in viable counts (CFU/g) of staphylococci with respect to the untreated control. The presence of bacteriocin also had a moderate negative effect on total lactic acid bacteria. AS-48(+) strain was developed well in the meat mixture, producing sufficient amounts of AS-48 (to a maximum of 76-88 arbitrary units/g) to control growth of staphylococci. The best result was achieved with a bacteriocinogenic strain inoculum of 10(7)CFU/g.