Ligilactobacillus murinus Strains Isolated from Mice Intestinal Tract: Molecular Characterization and Antagonistic Activity against Food-Borne Pathogens.

Considering the objectives of "One Health" and the Sustainable development Goals "Good health and well-being" for the development of effective strategies to apply against bacterial resistance, food safety dangers, and zoonosis risks, this project explored the isolation and identification of Lactobacillus strains from the intestinal tract of recently weaned mice; as well as the assessment of antibacterial activity against clinical and zoonotic pathogens. For molecular identification, 16S rRNA gene-specific primers were used and, via BLAST-NCBI, 16 Ligilactobacillus murinus, one Ligilactobacillus animalis, and one Streptococcus salivarius strains were identified and registered in GenBank after the confirmation of their identity percentage and the phylogenetic analysis of the 16 Ligilactobacillus murinus strains and their association with Ligilactobacillus animalis. The 18 isolated strains showed antibacterial activity during agar diffusion tests against Listeria monocytogenes ATCC 15313, enteropathogenic Escherichia coli O103, and Campylobacter jejuni ATCC 49943. Electrophoretic and zymographic techniques confirmed the presence of bacteriolytic bands with a relative molecular mass of 107 kDa and another of 24 kDa in Ligilactobacillus murinus strains. UPLC-MS analysis allowed the identification of a 107 kDa lytic protein as an N-acetylmuramoyl-L-amidase involved in cytolysis and considered a bacteriolytic enzyme with antimicrobial activity. The 24 kDa band displayed similarity with a portion of protein with aminopeptidase function. It is expected that these findings will impact the search for new strains and their metabolites with antibacterial activity as an alternative strategy to inhibit pathogens associated with major health risks that help your solution.

Organic trace minerals on productive performance, egg quality and immune response in Bovans White laying hens.

This study was performed in 360 laying Bovans White hens and aimed to evaluate the effect of Carbo-Amino-Phosphate-Chelates (CAFQ) as a mineral premix and its 30% reduction by compared to inorganic trace minerals (ITM) concerning its performance, egg quality, tibia breaking strength and immunity. Sixty-week-old hens were assigned into three treatments with 10 replicates of 12 birds each. Treatments were as follows: (a) (100% ITM), (b) (100% CAFQ) and (c) (70% CAFQ). Based on the results obtained during 12 weeks, under the current test conditions, improved yolk colour (p < 0.05); shell breaking strength (p < 0.05); and storage time (p < 0.05) were observed in hens fed with a diet reduced by 70% in CAFQ inclusion showed similar performance behaviour and better egg breaking strength regarding with ITM, suggesting the possibility of minimizing the inclusion of trace minerals in laying hen diets improving environmental impact, in contrast to the use of inorganic sources of minerals.

Nanostructured chitosan/propolis formulations: characterization and effect on the growth of Aspergillus flavus and production of aflatoxins.

A great diversity of agricultural products is susceptible to contamination caused by Aspergillus flavus. To reduce fungal contamination, the application of natural products has been proposed, including chitosan and propolis, due to its broad and recognized antimicrobial activity on several microorganisms. Currently, the application of nanotechnology allows for a greater activity to be more reactive and efficient. The objectives of this research were to characterize by TEM and Z potential some of the studied nanoparticles and to determine the in vitro antifungal activity of the formulations and the production of aflatoxins of the treated fungus. For this, individual treatments and different nanoformulations were elaborated by varying the percentage of the components such as chitosan solution, chitosan nanoparticles, an extract of propolis, nanoparticles of propolis, glycerol and canola oil. The final concentrations of the formulations were of 20%, 30% and 40% and the control consisted of Czapeck-dox agar medium. TEM micrographies showed a spherical morphology in a range of 2.3-3.0 nm with values of Z potential from 18.5 to 116.2 nm. Compared to the untreated fungus, the highest effect was seen in the parameter of spore germination, since inhibition was of c. a. 97% corresponding to the formulation containing chitosan + propolis nanoparticles + chitosan nanoparticles + propolis extract at the highest concentration of 40%. At this same concentration, the production of aflatoxins was 100% inhibited with the treatment with chitosan at 1%. Since these results are under carefully controlled conditions, further research should be extended to different fruit and vegetables affected by this fungus.

Ameliorative Effects of Neutral Electrolyzed Water on Growth Performance, Biochemical Constituents, and Histopathological Changes in Turkey Poults during Aflatoxicosis.

Different in vitro and in silico approaches from our research group have demonstrated that neutral electrolyzed water (NEW) can be used to detoxify aflatoxins. The objective of this investigation was to evaluate the ability of NEW to detoxify B-aflatoxins (AFB1 and AFB2) in contaminated maize and to confirm detoxification in an in vivo experimental model. Batches of aflatoxin-contaminated maize were detoxified with NEW and mixed in commercial feed. A total of 240 6-day-old female large white Nicholas-700 turkey poults were randomly divided into four treatments of six replicates each (10 turkeys per replicate), which were fed ad libitum for two weeks with the following dietary treatments: (1) control feed containing aflatoxin-free maize (CONTROL); (2) feed containing the aflatoxin-contaminated maize (AF); (3) feed containing the aflatoxin-contaminated maize detoxified with NEW (AF + NEW); and (4) control feed containing aflatoxin-free maize treated with NEW (NEW). Compared to the control groups, turkey poults of the AF group significantly reduced body weight gain and increased feed conversion ratio and mortality rate; whereas turkey poults of the AF + NEW group did not present significant differences on productive parameters. In addition, alterations in serum biochemical constituents, enzyme activities, relative organ weight, gross morphological changes and histopathological studies were significantly mitigated by the aflatoxin-detoxification procedure. From these results, it is concluded that the treatment of aflatoxin-contaminated maize with NEW provided reasonable protection against the effects caused by aflatoxins in young turkey poults.

Efecto de la adición de ácido cítrico sobre la degradación de las aflatoxinas y las propiedades funcionales de productos extrudidos de sorgo / Effect of citric acid addition on aflatoxin degradation and on functional properties of extruded sorghum products / Efeito da adição de ácido cítrico sobre a degradação das aflatoxinas e as propriedades funcionais de produtos extrudidos de sorgo

Harina de sorgo contaminada con aflatoxinas (AFB1+AFB2) a una concentración total de 140µg·kg-1, fue sometida a extrusión termoplástica usando un perfil de temperatura de 60-80-100ºC. El contenido de humedad (CH) de la harina fue ajustado a 20, 25 y 30% mediante la adición de diferentes cantidades de ácido cítrico (AC) acuoso a concentraciones de 0; 0,5; 1; 2; 4 y 8N. El perfil de temperatura, en combinación con el CH y la concentración de AC, afectaron significativamente la cantidad de aflatoxinas recuperadas en los extrudidos; el máximo porcentaje de eliminación fue de 35,5%. En los extrudidos procesados con altos CH y concentraciones de AC se encontraron los valores más altos en la diferencia de color (DE), así como los valores más bajos en viscosidad. Los índices de expansión fueron favorecidos a mayores CH y altas concentraciones de AC. Los mayores valores de fuerza al corte se encontraron en los extrudidos elaborados con un mayor CH; sin embargo, no se presentaron diferencias significativas para las diferentes concentraciones de AC evaluadas. Empleando condiciones moderadas durante la extrusión en combinación con altas concentraciones de AC se obtuvieron extrudidos con aceptable pH, color, viscosidad, así como con buenas propiedades funcionales y texturales.

Sorghum flour contaminated by aflatoxin at a concentration of 140µg·kg-1 was extrusion-cooked in a single screw extruder with a temperature profile of 60-80-100°C. The moisture content (MC) of the flour was adjusted to 20, 25 and 30% by adding aqueous citric acid (CA) at concentrations of 0, 0.5, 1, 2, 4 and 8N. The temperature profile in combination with the MC and CA concentrations significantly affected the extent of aflatoxin reduction in the extruded sorghum; the maximum percentage of aflatoxin reduction was 35.5%. In extruded sorghum prepared with high MC and CA concentrations, the highest values in color difference (DE) and smallest values of viscosity were found. The expansion index was favored by higher MC and CA concentrations. The highest cutting force values were found in extruded products prepared with a higher MC, but there were no significant differences between those values and the different CA concentrations evaluated. Using moderate extrusion conditions in combination with higher concentrations of CA, products were obtained with acceptable pH, color, viscosity, as well as with good textural and functional properties.

Farinha de sorgo contaminada com aflatoxinas (AFB1+AFB2) na concentração total de 140µg·kg-1, foi submetida ao proceso de extrusão termoplástica usando um perfil de temperatura de 60-80-100ºC. O conteúdo de umidade (CH) da farinha foi ajustado a 20, 25 e 30% mediante a adição de diferentes quantidades de ácido cítrico (AC) aquoso em concentrações de 0; 0,5; 1; 2; 4 e 8N. O perfil de temperatura, combinado com o CH e a concentração de AC, afetaram significativamente a quantidade de aflatoxinas recuperadas nos extrudidos; a máxima porcentagem de eliminação foi de 35,5%. Nos extrudidos procesados com alto CH e concentração de AC se encontraram os valores mais altos na diferença de cor (DE), assim como os valores mais baixos em viscosidade. Os índices de expansão foram favorecidos a maior CH e altas concentrações de AC. Os maiores valores de força ao corte se encontraram nos extrudidos elaborados com maior CH; no entanto, não se apresentaram diferenças significativas para as diferentes concentrações de AC avaliadas. Utilizando condiciones moderadas durante a extrusão em combinação com altas concentrações de AC se obtiveram extrudidos com aceitável pH, cor, viscosidade, assim como com boas propriedades funcionais e texturais.

[Evaluation of the cytotoxicity of aflatoxin and fumonisin in swine intestinal cells]. / Evaluación de la citotoxicidad de la aflatoxina y la fumonisina en células intestinales de porcino.

The coexistence of the aflatoxin (AFB) and fumonisin (FB) has been widely documented in many parts of the world. However, few studies describing the synergy effect of both mycotoxins in vivo and/or in vitro are available. The objective of this study consisted on evaluating the effect of AFB and FB on the morphology, the capacity of cellular proliferation, cytotoxicity and interleukin-8 (IL-8) synthesis in a porcine intestinal epithelial cell line (IPEC-1). Concerning to the cellular morphology it was only affected in the concentrations higher of AFB (50 microM) and FB (500 microM). However, the cellular proliferation, the cellular damage and synthesis of IL-8 they were affected when present in combination the AFB/FB (1.3/3.7; 2/3.7 and 5/10 microM respectively) with that showed by the individual effect of similar concentrations of these mycotoxins (p < 0.05). Our data indicate that the combination of AFB/FB in low concentrations showed a synergy effect, altering the cellular morphophysiology, which can imply in vivo the entrance of other toxins or biological agents for alteration of the intestinal barrier impacting negatively in the human or animal health.