Discovery of antimicrobial peptides clostrisin and cellulosin from Clostridium: insights into their structures, co-localized biosynthetic gene clusters, and antibiotic activity.

Antimicrobial resistance presents a substantial threat to global public health, demanding urgent attention and action. This study focuses on lanthipeptides, ribosomally encoded peptides that display significant structural diversity and hold promising potential as antibiotics. Genome mining was employed to locate biosynthetic gene clusters (BGCs) containing class II lanthipeptide synthetases encoded by lanM genes. A phylogenetic study analyzing homologous sequences of functional LanM sequences revealed a unique evolutionary clade of 17 LanM proteins associated with 12 Clostridium bacterial genomes. In silico exploration identified nine complete BGCs, including one super-cluster containing two co-localized operons from Clostridium cellulovorans 743B, that encode for two new peptides named clostrisin and cellulosin. Each operon was heterologously expressed in Escherichia coli. Molecular weights associated with the expected post-translational modifications of the purified lanthipeptide were confirmed by MS-MS/MS analysis for cellulosin, while clostrisin was not post-translationally modified. Both peptides demonstrated antimicrobial activity against multidrug-resistant bacteria, such as a clinical strain of Staphylococcus epidermidis MIQ43 and Pseudomonas aeruginosa PA14. This is the first report of lanthipeptides from the Clostridium genus produced with its native biosynthetic machinery, as well as chemically and biologically characterized. This study showcases the immense potential of genome mining in identifying new RiPP synthetases and associated bioactive peptides.

Implementing an Antimicrobial Stewardship Program in an Oncology Center in Lima, Peru: A Model for Low- and Middle-Income Countries.

Our center launched the first antimicrobial stewardship program in Peru. From 2016 to 2023, the proportion of antimicrobial prescriptions audited increased from 60% to 95%, and 65% to 95% of recommendations were accepted. Vancomycin and meropenem use dropped by 95% and 84%, respectively. The proportion of recommendations for surgical prophylaxis exceeded 90%.

Competitive exclusion products as an antimicrobial alternative to control Salmonella Heidelberg in broilers.

Intestinal infections caused by non-typhoidal Salmonella spp., along with antimicrobial resistance spread are a major food safety concern worldwide. Here, we evaluate the potential of competitive exclusion products developed by anaerobic or aerobic conditions to control systemic infection, cecal colonization, fecal excretion, and improve the intestinal health in broilers challenged by Salmonella Heidelberg (SH). A total of 105 day-old chickens were randomly distributed into three experimental groups: A (untreated control), B (treated with anaerobic culture), and C (treated with aerobic culture). During 21 days, morphometric parameters of the small intestine were analyzed using microscopy, fecal excretions by cloacal swabs, systemic infection, and cecal colonization by colony-forming unit counts (CFU/g). The results indicated the lowest number of positive swabs (45.33%) recovered from Group C, followed by Group B (71.8%) and Group A (85.33%). The bacterial enumeration revealed the lowest amounts in Group C at the necropsy realized in 5-, 7-, and 14-days post-infection (DPI) (P = 0.0010, P = 0.0048, and P = 0.0094, respectively). Statistical differences between intestinal morphometrics were observed in the Group C at 21 DPI. Our results suggest that the product developed under aerobic conditions can improve intestinal health, protecting birds against SH.

In vitro analysis of postbiotic antimicrobial activity against Candida Species in a minimal synthetic model simulating the gut mycobiota in obesity.

Gut fungal imbalances, particularly increased Candida spp., are linked to obesity. This study explored the potential of Lactiplantibacillus plantarum cell-free extracts (postbiotics) to modulate the growth of Candida albicans and Candida kefyr, key members of the gut mycobiota. A minimal synthetic gut model was employed to evaluate the effects of Lactiplantibacillus plantarum postbiotics on fungal growth in mono- and mixed cultures. Microreactors were employed for culturing, fungal growth was quantified using CFU counting, and regression analysis was used to evaluate the effects of postbiotics on fungal growth. Postbiotics at a concentration of 12.5% significantly reduced the growth of both Candida species. At 24 h, both C. albicans and C. kefyr in monocultures exhibited a decrease in growth of 0.11 log CFU/mL. In contrast, mixed cultures showed a more pronounced antifungal effect, with C. albicans and C. kefyr reductions of 0.62 log CFU/mL and 0.64 log CFU/mL, respectively. Regression analysis using the Gompertz model supported the antifungal activity of postbiotics and revealed species-specific differences in growth parameters. These findings suggest that L. plantarum postbiotics have the potential to modulate the gut mycobiota by reducing Candida growth, potentially offering a therapeutic approach for combating fungal overgrowth associated with obesity.

Subinhibitory concentrations of nisin enhance virulence gene expression in Staphylococcus aureus and increase mortality in Galleria mellonella.

Alternative strategies for controlling Staphylococcus aureus and other pathogens have been continuously investigated, with nisin, a bacteriocin widely used in the food industry as a biopreservative, gaining increasing attention. In addition to its antimicrobial properties, bacteriocins have significant effects on genome functionality even at inhibitory concentrations. This study investigated the impact of subinhibitory concentrations of nisin on S. aureus. Culturing in the presence of 0.625 µmol l-1 nisin, led to the increased relative expression of hla, saeR, and sarA, genes associated with virulence while expression of the sea gene, encoding staphylococcal enterotoxin A (SEA), decreased. In an in vivo experiment, Galleria mellonella larvae inoculated with S. aureus cultured in the presence of nisin exhibited 97% mortality at 72 h post-infection, compared to over 40% of larvae mortality in larvae infected with S. aureus. A comprehensive understanding of the effect of nisin on the transcriptional response of virulence genes and the impact of these changes on the virulence of S. aureus can contribute to assessing the application of this bacteriocin in food and medical contexts.

Metagenomic Analysis of Rhizospheric Bacterial Community of Citrus Trees Expressing Phloem-Directed Antimicrobials.

Huanglongbing, also known as citrus greening, is currently the most devastating citrus disease with limited success in prevention and mitigation. A promising strategy for Huanglongbing control is the use of antimicrobials fused to a carrier protein (phloem protein of 16 kDa or PP16) that targets vascular tissues. This study investigated the effects of genetically modified citrus trees expressing Citrus sinensis PP16 (CsPP16) fused to human lysozyme and ß-defensin-2 on the soil microbiome diversity using 16S amplicon analysis. The results indicated that there were no significant alterations in alpha diversity, beta diversity, phylogenetic diversity, differential abundance, or functional prediction between the antimicrobial phloem-overexpressing plants and the control group, suggesting minimal impact on microbial community structure. However, microbiota diversity analysis revealed distinct bacterial assemblages between the rhizosphere soil and root environments. This study helps to understand the ecological implications of crops expressing phloem-targeted antimicrobials for vascular disease management, with minimal impact on soil microbiota.

Moderate and serious adverse reactions to antimicrobials among hospitalized children: A systematic review.

AIMS: This systematic review aimed to investigate the occurrence of moderate and severe adverse drug reactions (ADRs) to antimicrobials among hospitalized children. METHODS: The PubMed/Medline, Cochrane Library, Embase, Web of Science, Scopus, Lilacs and CINAHL databases were searched in April 2023 to systematically review the published data describing the characteristics of moderate and severe ADRs to antimicrobials among hospitalized children. The search was carried out without date restrictions, up to the search date (April, 2023). RESULTS: At the end of the selection process, 30 articles met the inclusion criteria. Cutaneous reactions were the primary serious clinical manifestations in most articles (19/30), followed by erythema multiforme (71 cases), Stevens-Johnson syndrome (72 cases), and toxic epidermal necrolysis (22 cases). The main antimicrobials involved in moderate and severe ADRs were penicillins, cephalosporins and sulfonamides. Regarding the primary outcomes, 30% (9/30) of the articles reported deaths, and 46.7% (14/30) of studies reported increased lengths of hospital stay, need for intensive care, and transfer to another hospital. Regarding the main interventions, 10% (3/30) of the articles mentioned greater monitoring, suspension, medication substitution or prescription of specific medications for the symptomatology. CONCLUSIONS: The findings of this review could be used to identify areas for improvement and help health professionals and policymakers develop strategies. In addition, we emphasize the importance of knowing about ADRs so that there is adequate management to avoid undesirable consequences.

Synthesis and Isolation of Phenol- and Thiol-Derived Epicatechin Adducts Prepared from Avocado Peel Procyanidins Using Centrifugal Partition Chromatography and the Evaluation of Their Antimicrobial and Antioxidant Activity.

Polyphenols from agro-food waste represent a valuable source of bioactive molecules that can be recovered to be used for their functional properties. Another option is to use them as starting material to generate molecules with new and better properties through semi-synthesis. A proanthocyanidin-rich (PACs) extract from avocado peels was used to prepare several semi-synthetic derivatives of epicatechin by acid cleavage in the presence of phenol and thiol nucleophiles. The adducts formed by this reaction were successfully purified using one-step centrifugal partition chromatography (CPC) and identified by chromatographic and spectroscopic methods. The nine derivatives showed a concentration-dependent free radical scavenging activity in the DPPH assay. All compounds were also tested against a panel of pathogenic bacterial strains formed by Listeria monocytogenes (ATCC 7644 and 19115), Staphylococcus aureus (ATCC 9144), Escherichia coli (ATCC 11775 and 25922), and Salmonella enterica (ATCC 13076). In addition, adducts were tested against two no-pathogenic strains, Limosilactobacillus fermentum UCO-979C and Lacticaseibacillus rhamnosus UCO-25A. Overall, thiol-derived adducts displayed antimicrobial properties and, in some specific cases, inhibited biofilm formation, particularly in Listeria monocytogenes (ATCC 7644). Interestingly, phenolic adducts were inactive against all the strains and could not inhibit its biofilm formation. Moreover, depending on the structure, in specific cases, biofilm formation was strongly promoted. These findings contribute to demonstrating that CPC is a powerful tool to isolate new semi-synthetic molecules using avocado peels as starting material for PACc extraction. These compounds represent new lead molecules with antioxidant and antimicrobial activity.

Dictyostelium discoideum-assisted pharmacognosy of plant resources for discovering antivirulence molecules targeting Klebsiella pneumoniae.

The rise of antibiotic-resistant bacterial strains represents an important challenge for global health, underscoring the critical need for innovative strategies to confront this threat. Natural products and their derivatives have emerged as a promising reservoir for drug discovery. The social amoeba Dictyostelium discoideum is a potent model organism in this effort. Employing this invertebrate model, we introduce a novel perspective to investigate natural plant extracts in search of molecules with potential antivirulence activity. Our work established an easy-scalable developmental assay targeting a virulent strain of Klebsiella pneumoniae, with Helenium aromaticum as the representative plant. The main objective was to identify tentative compounds from the Helenium aromaticum extract that attenuate the virulence of K. pneumoniae virulence without inducing cytotoxic effects on amoeba cells. Notably, the methanolic root extract of H. aromaticum fulfilled these prerequisites compared to the dichloromethane extract. Using UHPLC Q/Orbitrap/ESI/MS/MS, 63 compounds were tentatively identified in both extracts, 47 in the methanolic and 29 in the dichloromethane, with 13 compounds in common. This research underscores the potential of employing D. discoideum-assisted pharmacognosy to discover new antivirulence agents against multidrug-resistant pathogens.

Carvacrol and thymol effect in vapor phase on Escherichia coli and Salmonella serovar Typhimurium growth inoculated in a fresh salad.

This study aimed to evaluate the antimicrobial effect of thymol and carvacrol in inhibiting Escherichia coli and Salmonella serovar Typhimurium inoculated on a fresh green salad through the vapor phase. A film-forming solution was prepared by dissolving starch, sorbitol, and variying concentrations of carvacrol, thymol, and a mixture of both. The film-forming solution containing the respective antimicrobial agent was then added lid, which was sealed rigidly and hermetically to achieve different concentrations (105 mg/L of air of carvacrol, 105 mg/L of air of thymol, and a mixture of 52 mg/L of air of carvacrol and 52 mg/L of air of thymol). Each active package contained fresh green salad inoculated with E. coli or Salmonella serovar Typhimurium. The active packages were then sealed and refrigerated at a temperature of 6 °C for 48 h. Growth/inhibition curves were modelled using the Weibull equation, and consumer acceptance was evaluated. Carvacrol can reduce up to 0.5 log-cycles, while thymol can reach almost 1 log cycle. Blending the components with half the concentration has a synergistic effect, inhibiting up to 2.5 log cycles. Consumer ratings revealed no significant differences between the packages. However, the average score was 5.4 on a 9-point hedonic scale, evaluators' comments did not indicate dislike or a strong taste characteristic of thymol and carvacrol.

Utilization of Whey for Eco-Friendly Bio-Preservation of Mexican-Style Fresh Cheeses: Antimicrobial Activity of Lactobacillus casei 21/1 Cell-Free Supernatants (CFS).

Using whey, a by-product of the cheese-making process, is important for maximizing resource efficiency and promoting sustainable practices in the food industry. Reusing whey can help minimize environmental impact and produce bio-preservatives for foods with high bacterial loads, such as Mexican-style fresh cheeses. This research aims to evaluate the antimicrobial and physicochemical effect of CFS from Lactobacillus casei 21/1 produced in a conventional culture medium (MRS broth) and another medium using whey (WB medium) when applied in Mexican-style fresh cheese inoculated with several indicator bacteria (Escherichia coli, Salmonella enterica serovar Typhimurium, Staphylococcus aureus, and Listeria monocytogenes). The CFSs (MRS or WB) were characterized for organic acids concentration, pH, and titratable acidity. By surface spreading, CFSs were tested on indicator bacteria inoculated in fresh cheese. Microbial counts were performed on inoculated cheeses during and after seven days of storage at 4 ± 1.0 °C. Moreover, pH and color were determined in cheeses with CFS treatment. Lactic and acetic acid were identified as the primary antimicrobial metabolites produced by the Lb. casei 21/1 fermentation in the food application. A longer storage time (7 days) led to significant reductions (p < 0.05) in the microbial population of the indicator bacteria inoculated in the cheese when it was treated with the CFSs (MRS or WB). S. enterica serovar Typhimurium was the most sensitive bacteria, decreasing 1.60 ± 0.04 log10 CFU/g with MRS-CFS, whereas WB-CFS reduced the microbial population of L. monocytogenes to 1.67 log10 CFU/g. E. coli and S. aureus were the most resistant at the end of storage. The cheese's pH with CFSs (MRS or WB) showed a significant reduction (p < 0.05) after CFS treatment, while the application of WB-CFS did not show greater differences in color (ΔE) compared with MRS-CFS. This study highlights the potential of CFS from Lb. casei 21/1 in the WB medium as an ecological bio-preservative for Mexican-style fresh cheese, aligning with the objectives of sustainable food production and guaranteeing food safety.

Multi-locus sequence typing, antimicrobials resistance and virulence profiles of Salmonella enterica isolated from bovine carcasses in Minas Gerais state, Brazil.

The aim of this study was to identify virulence and antimicrobial resistance profiles and determine the sequence type (ST) by multilocus sequence typing (MLST) of Salmonella enterica isolates from bovine carcasses from slaughterhouse located in Minas Gerais state, Brazil, and its relationship with bovine isolates obtained on the American continent based on sequence type profile. The MLST results were compared with all Salmonella STs associated with cattle on American continent, and a multi-locus sequence tree (MS tree) was built. Among the 17 S. enterica isolates, five ST profiles identified, and ST10 were the most frequent, grouping seven (41.2%) isolates. The isolates presented 11 different profiles of virulence genes, and six different antibiotics resistance profiles. The survey on Enterobase platform showed 333 Salmonella STs from American continent, grouped into four different clusters. Most of the isolates in the present study (13/17), were concentrated in a single cluster (L4) composed by 74 STs. As a conclusion, five different STs were identified, with ST10 being the most common. The isolates showed great diversity of virulence genes and antibiotics resistance profiles. Most of the isolates of this study were grouped into a single cluster composed by 74 STs formed by bovine isolates obtained on the American continent.

Probiotics for Aquaculture: Hope, Truth, and Reality.

The use of microorganisms as beneficial crops for human and animal health has been studied for decades, and these microorganisms have been in practical use for quite some time. Nowadays, in addition to well-known examples of beneficial properties of lactic acid bacteria, bifidobacteria, selected Bacillus spp., and yeasts, there are several other bacteria considered next-generation probiotics that have been proposed to improve host health. Aquaculture is a rapidly growing area that provides sustainable proteins for consumption by humans and other animals. Thus, there is a need to develop new technologies for the production practices associated with cleaner and environment-friendly approaches. It is a well-known fact that proper selection of the optimal probiotics for use in aquaculture is an essential step to ensure effectiveness and safety. In this critical review, we discuss the evaluation of host-specific probiotics in aquaculture, challenges in using probiotics in aquaculture, methods to improve the survival of probiotics under different environmental conditions, technological approach to improving storage, and delivery along with possible negative consequences of using probiotics in aquaculture. A critical analysis of the identified challenges for the use of beneficial microbes in aquaculture will help in sustainable aquafarming, leading to improved agricultural practices with a clear aim to increase protein production.

Prevalence and antibiotic resistance in bacterial isolates of dogs with ulcerative keratitis in São Paulo State, Brazil.

OBJECTIVE: Identify microorganisms present in canine eyes affected by ulcerative keratitis and assess its resistance profile to available antimicrobial drugs. METHODS: Samples were collected from 88 canine eyes that exhibited ulcerative keratitis. They were identified using MALDI-TOF and subjected to antimicrobial susceptibility testing by disk diffusion. RESULTS: Among the assessed subjects, brachycephalic dogs accounted for 74.48% (50/83) of the evaluated canines. Among the 88 evaluated eyes, 90.9% (80/88) showed positive cultures, with 11.33% (10/88) of the samples isolating more than one species of bacteria. Of all bacterial isolates identified (90), Gram-positive bacteria accounted for 63.33% (57/90), while Gram-negative bacteria constituted 36.66% (33/90), with predominance of Staphylococcus spp. at 35.55% (32/90) being, Staphylococcus pseudintermedius at 68.75% (22/32), and Pseudomonas aeruginosa at 15.55% (14/90), respectively. Staphylococcus spp. exhibited resistance to penicillin (89.29%), sulfadiazine and trimethoprim (60.71%), and tetracycline (67.86%), while doxycycline (88.89%), cefotaxime (85.71%), chloramphenicol (82.14%), gentamicin, and moxifloxacin (78.57%) showed the highest sensitivity rates. Pseudomonas aeruginosa displayed sensitivity (100%) to gentamicin and imipenem, and resistance (8.33%) to norfloxacin, ciprofloxacin, and cefepime. Similarly, the Enterobacteriaceae family showed higher sensitivity to amikacin and gentamicin (88.89%), imipenem (88.24%), and levofloxacin (87.5%), with pronounced resistance to amoxicillin-clavulanate (50%) and cefazolin (47.06%). This highlights multiresistance in 23.33% (21/90) of the isolates. CONCLUSIONS: The most isolated species in canine ulcerative keratitis are S. pseudintermedius and P. aeruginosa. However, other species were also isolated, demonstrating diversity in ocular microbiota infection. There is a high-rate multidrug resistance associated with canine ulcerative keratitis. Nevertheless, these strains exhibited sensitivity to antimicrobials commonly used in veterinary ophthalmology.

Active Films of Cassava Starch Incorporated with Carvacrol Nanocapsules.

The synthesis of active films with natural antimicrobials from renewable sources offers an alternative to conventional non-biodegradable packaging and synthetic additives. This study aimed to develop cassava starch films with antimicrobial activity by incorporating either free carvacrol or chia mucilage nanocapsules loaded with carvacrol (CMNC) and assess their impact on the physical, mechanical, and barrier properties of the films, as well as their efficacy against foodborne pathogens. The addition of free carvacrol led to a reduction in mechanical properties due to its hydrophobic nature and limited interaction with the polymeric matrix. Conversely, CMNC enhanced elongation at break and reduced light transmission, with a more uniform distribution in the polymeric matrix. Films containing 8% carvacrol exhibited inhibitory effects against Salmonella and Listeria monocytogenes, further potentiated when encapsulated in chia mucilage nanocapsules. These findings suggest that such films hold promise as active packaging materials to inhibit bacterial growth, ensuring food safety and extending shelf life.

Resistencia antimicrobiana de Enterobacterias causante de infección del tracto urinario en pacientes ambulatorios / Antimicrobial resistance of Enterobacteriaceae causing urinary tract infection in ambulatory patients / Resistência antimicrobiana de Enterobacteriaceae causadora de infecção do trato urinário em pacientes ambulatoriais

Las infecciones del tracto urinario son consideradas un problema de salud a nivel hospitalario y comunitario por el aumento de bacterias resistentes a los antibióticos. Objetivo: Analizar el patrón de susceptibilidad y resistencia antimicrobiana de Enterobacterias causante de infección del tracto urinario. Métodos: Se aplicó una investigación descriptiva de diseño documental. La población fue de 672 registros de urocultivos positivos, recopilados de la base de datos del Laboratorio San Pablo en el periodo 2021-2022. Para su tabulación y análisis los datos obtenidos fueron procesados en el Software SPSS versión 25.0. Resultados: Las ITU se presentan con mayor frecuencia en el género femenino 86,5%. El grupo etario con más afección es la edad adulta 50,4%. El agente etiológico con mayor incidencia fue Escherichia coli 75,74%, Citrobacter Freundii 8,93%, Klebsiella spp 6,10%. La producción de BLEE como mecanismo de resistencia predominaron en las cepas de E.coli y Klebsiella spp. Se encontró un mayor porcentaje de resistencia para Ampicilina y SXT. Los antibióticos con mejor sensibilidad destacaron nitrofurantoína, fosfomicina. Conclusión: La especie con mayor aislamiento, implicada en la etiología de infecciones urinarias sigue siendo E.coli con una sensibilidad alta para nitrofurantoína y fosfomicina.

Urinary tract infections are considered a health problem at hospital and community level due to the increase of antibiotic resistant bacteria. Objective: To analyze the pattern of susceptibility and antimicrobial resistance of Enterobacteriaceae causing urinary tract infection. Methods: A descriptive research of documentary design was applied. The population was 672 records of positive urine cultures, collected from the San Pablo Laboratory database in the period 2021-2022. For tabulation and analysis, the data obtained were processed in SPSS software version 25.0. Results: UTIs occur more frequently in females 86.5%. The age group with the highest incidence was adulthood 50.4%. The etiological agent with the highest incidence was Escherichia coli 75.74%, Citrobacter Freundii 8.93%, Klebsiella spp 6.10%. The production of BLEE as a mechanism of resistance predominated in the strains of E.coli and Klebsiella spp. A higher percentage of resistance was found for Ampicillin and SXT. The antibiotics with the best sensitivity were nitrofurantoin and fosfomycin. Conclusion: The species with the highest isolation, implicated in the etiology of urinary tract infections, continues to be E.coli with a high sensitivity to nitrofurantoin and fosfomycin.

As infecções do trato urinário são consideradas um problema de saúde a nível hospitalar e comunitário devido ao aumento de bactérias resistentes aos antibióticos. Objetivo: Analisar o padrão de suscetibilidade e resistência antimicrobiana das Enterobacteriaceae causadoras de infecções do trato urinário. Métodos: Foi aplicada uma metodologia de investigação documental descritiva. A população foi de 672 registros de culturas de urina positivas, coletados do banco de dados do Laboratório San Pablo no período de 2021-2022. Para tabulação e análise, os dados obtidos foram processados no software SPSS versão 25.0 Resultados: As ITUs ocorreram com maior frequência no sexo feminino 86,5%. A faixa etária com maior incidência foi a adulta 50,4%. O agente etiológico com maior incidência foi a Escherichia coli 75,74%, Citrobacter Freundii 8,93%, Klebsiella spp 6,10%. A produção de BLEE como mecanismo de resistência predominou em E. coli e Klebsiella spp. Foi encontrada uma maior percentagem de resistência para a ampicilina e o SXT. Os antibióticos com melhor sensibilidade foram a nitrofurantoína e a fosfomicina. Conclusão: A espécie com maior isolamento, implicada na etiologia das infecções do trato urinário, continua a ser a E. coli com uma elevada sensibilidade à nitrofurantoína e à fosfomicina.

Control of Escherichia coli in Poultry Using the In Ovo Injection Technique.

Pathogens, such as Escherichia coli (E. coli), have been identified as significant causes of poultry mortality. Poultry can serve as potential sources of E. coli transmission, even when asymptomatic, posing a substantial threat to food safety and human health. The in ovo administration of antimicrobials is crucial for preventing and/or effectively combating acute and chronic infections caused by poultry pathogens. To achieve this goal, it is critical that antimicrobials are properly injected into embryonic fluids, such as the amnion, to reach target tissues and trigger robust antimicrobial responses. Several protocols based on antimicrobials were evaluated to meet these requirements. This review analyzed the impacts of antimicrobial substances injected in ovo on the control of E. coli in poultry. The reduction in infection rates, resulting from the implementation of in ovo antimicrobials, combined with efforts aimed at hygienic-sanitary action plans in poultry sheds, reinforces confidence that E. coli can be contained before causing large scale damage. For example, antimicrobial peptides and probiotics have shown potential to provide protection to poultry against infections caused by E. coli. Issues related to the toxicity and bacterial resistance of many synthetic chemical compounds represent challenges that need to be overcome before the commercial application of in ovo injection protocols focused on microbiological control.

Preventing microbe colonization on avocado (Persea nubigena var. guatemalensis) through metabiotic treatment, a promising postharvest safety improvement.

Introduction: Lactic acid bacteria (LAB) produce various metabolites (i.e. metabiotics) with inhibitory capacity towards harmful foodborne pathogens. Methods: This study aimed to design several antimicrobial formulations based on metabiotics obtained from different native LAB species (Lactobacillus pentosus UTNGt5, Lactococcus lactis UTNGt28, and Weissella cibaria UTNGt21O) and to detect the possible mode of action towards two multidrug resistant Staphylococcus spp. strains isolated from avocado (Persea nubigena var. guatemalensis) fruits. Additionally, the formulation with the highest inhibitory activity was tested ex vitro on avocados at the immature (firm) ripeness stage to evaluate their effect on microorganisms' growth and fruit quality attributes post-harvest. Results and discussion: Out of the top five formulations showing the highest bactericidal effect in vitro at their minimum inhibitory concentration (1 x MIC) on both Staphylococcus spp. targets one candidate annotated P11 (consisting of UTNGt21O and UTNGt28; 1:3, v/v) was selected. Co-cultivation of Staphylococcus strains with P11 formulation results in cell viability reduction by 98%, by impairing the integrity of the cell membrane inducing cytoplasm molecule content leakage, protein profile changes, and finally bacterial death. Even though the total coliforms, Staphylococcus spp., Enterobacter spp., molds, and yeasts counts were not fully eliminated by day 13 of storage, a statistically significant reduction (p < 0.05) in viable cell counts were observed by day 8 upon the P11 treatment compared with non-treated control (C) and treated with a commercial disinfectant (T1) samples, suggesting that P11 formulation inhibited microbial colonization during storage. Likewise, no visible dark spots were observed on the mesocarp (pulp) upon the treatment with P11, whereas T1 and C fruits showed greater dark spots on the pulp as indicative of damage. The quality attributes, such as pH, total soluble solids, total titratable acidity, antioxidant capacity, and total polyphenol content, were not affected by the treatment. Principal Component Analysis (PCA) conducted on these five variables showed a clear separation of samples according to the maturity stage regardless of the treatment. Conclusion: These results suggest that the active metabolites from LAB strains might create a barrier between the exocarp and mesocarp, inhibiting the microorganisms colonization, reducing fruit damage, and lengthening the fruit quality and safety after harvest.