Search Results (77)

Background/Objectives: The growing threat of antimicrobial resistance has intensified the search for alternative therapeutic approaches. Quorum sensing (QS) inhibition, which disrupts bacterial communication and virulence, represents a promising approach to mitigating infection. Given the complexity of the sponge holobiont, sponge-associated microorganisms may demonstrate QS inhibitory properties and serve as potential sources of novel anti-virulence agents. This study aimed to investigate the QS inhibitory potential of sponge-associated Bacillus species against Pseudomonas aeruginosa, a multidrug-resistant pathogen that relies on QS for virulence regulation. Methods: Ninety-eight bacterial isolates were obtained from seven intertidal South African sponges. Biosensor-based sandwich assays using Chromobacterium violaceum identified 15 isolates with putative QS inhibition (QSI) activity, including five classified as Bacillus species via 16S rRNA gene sequencing. Crude extracts from these isolates, cultivated in medium Mannitol (Mann) and medium 5294, were screened for their ability to inhibit QS-regulated virulence factors in P. aeruginosa. Results: Extracts, particularly from medium 5294, exhibited significant QSI activity without cytotoxic effects. The five most potent extracts, i.e., Bacillus mobilis SP2-AB7 (5294), Bacillus wiedmannii SP5-AB7 (Mann), B. mobilis SP2-AB7 (Mann), and Bacillus cereus SP1-AB4 (Mann and 5294), inhibited both Las- and Rhl-regulated virulence factors, including pyocyanin, pyoverdine, elastase, protease, rhamnolipid production, motility, and initial adhesion, achieving inhibition rates of up to 93% (p < 0.05). Molecular analysis confirmed the presence of the aiiA lactonase gene in key isolates, while GC-MS and FTIR profiling revealed medium-specific differences in metabolite production. Conclusions: Sponge-associated Bacillus species from KwaZulu-Natal exhibit robust QSI activity against P. aeruginosa, highlighting their potential as sources of alternative anti-virulence agents. Further characterization and in vivo validation are needed to assess their therapeutic application in combatting resistant infections. Full article

This research reports ruminal and fecal microbiota composition of lactating dairy cows enrolled in a study aimed at investigating the effects of a fermentation extract derived from Bacillus licheniformis (BLFE), monensin (Rumensin^®; R), and their interactions on feed efficiency (FE, FE = milk yield/DMI). In a completely randomized design, 48 Holstein cows at 108 ± 35 days in milk were matched for parity and assigned to monensin (0 or 17.6 g/kg of DM) and BLFE (0 or 166 mg/kg of DM) in a 2 × 2 factorial arrangement. Treatments were fed daily for 63 d, including a 21 d adaptation period followed by a 42 d measurement period (P2). On d 38 and d 39 of P2, rumen-fluid (RF) and fecal samples were collected. DNA from RF and feces was sequenced using 16S rRNA gene-amplicon sequencing on an Illumina MiSeq platform. Fecal and RF volatile fatty acid (VFA) concentrations were analyzed, and propionate/acetate (P: A) was determined. The BLFE increased milk yield (3.3 kg/d) and FE (1.20 to 1.28), when fed alone rather than with monensin, while monensin increased energy-corrected milk yield (2.5 kg/d, p < 0.05), regardless of the BLFE in the diet. The BLFE tended to increase ruminal Firmicutes/Bacteroidetes (F: B) when fed alone, while alpha and beta diversities remained unmodified. The BLFE increased the abundances of Bifidobacterium (p = 0.02) and Erysipelotrichaceae_UCG-002 (p = 0.01) in RF, whereas monensin increased and decreased the abundances of Oscillospirales_ge (p = 0.02) and an unclassified Clostridia genus (p = 0.03), respectively. The monensin-suppressed Clostridia were negatively associated with ruminal P: A (r = −0.66; p < 0.01) and feed efficiency (r = −0.30; p = 0.04). The BLFE and monensin interactively affected several fecal genera (p < 0.05), but they had negligible or weak correlations with fecal P: A and FE. Overall, the results showed the ability of dietary supplementations of monensin and BLFE to increase milk production performance and FE by modulating ruminal rather than lower-gut microbiota composition, this is predominantly attributed to the ratio between the Firmicutes and Bacteroidetes abundances in lactating dairy cows. Full article

This study investigated the isolation and formulation of a bacterial conditioner as a biostimulant for Triticum durum (durum wheat) under salinity stress. An Algerian alkaline–saline soil was sampled, characterized for its physical and chemical characteristics and its culturable and total microbial community (16S rRNA gene metabarcoding). Three bacterial strains showing high 16S rRNA gene similarity to Pseudomonas putida, Bacillus proteolyticus, and Niallia nealsonii were selected for their plant growth-promoting (PGP) traits under different salinity levels, including phosphate solubilisation (194 µg mL⁻¹), hormone production (e.g., gibberellin up to 56 µg mL⁻¹), and good levels of hydrocyanic acid, ammonia, and siderophores. N. nealsonii maintained high indole production under saline conditions, while B. proteolyticus displayed enhanced indole synthesis at higher salt concentrations. Siderophore production remained stable for P. putida and N. nealsonii, whereas for B. proteolyticus a complete inhibition was registered in the presence of salt stress. The consortium density and application were tested under controlled conditions using Medicago sativa as a model plant. The effective biostimulant formulation was tested on Triticum durum under greenhouse experiments. Bacterial inoculation significantly improved plant growth in the presence of salt stress. Root length increased by 91% at 250 mM NaCl. Shoot length was enhanced by 112% at 500 mM NaCl. Total chlorophyll content increased by 208% at 250 mM NaCl. The chlorophyll a/b ratio increased by 117% at 500 mM. Also, reduced amounts of plant extracts were necessary to scavenge 50% of radicals (−22% at 250 mM compared to the 0 mM control). Proline content increased by 20% at both 250 mM and 500 mM NaCl. These results demonstrate the potential of beneficial bacteria as biostimulants to mitigate salt stress and enhance plant yield in saline soils. Full article

This study assessed the viable and culturable microbial diversity that remained on equipment surfaces after hygiene procedures in Brazilian poultry and fish slaughterhouses. Food-contact surface samples were collected using sterile swabs in poultry (n = 50) and fish (Oreochromis niloticus, n = 50) slaughterhouses. The swab samples were used to prepare culture plates to recover viable and culturable cells. The grown plates were washed, and the total DNA of the cell suspension was extracted with a commercial kit. Sequencing of the total DNA extracted from cultures was targeted at the V3 and V4 regions of the 16S rRNA. DNA reads were analyzed by QIIME2 software, with results expressed in relative frequency (%RF). Alpha and beta diversity indexes were analyzed considering the spots of sample collection, type of industry, surfaces (smooth or modular), and materials (polypropylene, stainless steel, or polyurethane). The results showed that in the poultry slaughterhouse, the most abundant genera were Acinetobacter (27.4%), Staphylococcus (7.7%), and Pseudomonas (5.3%), while for the fish slaughterhouse, there was a higher abundance of Staphylococcus (27.7%), Acinetobacter (17.2%), and Bacillus (12.5%). Surface characteristics influenced the microbial diversity, with Acinetobacter spp. dominating modular surfaces and Staphylococcus spp. prevailing on smooth surfaces. The results obtained indicate there is an important resident microbiota that persists even after hygiene processes, and surface-specific cleaning strategies should be developed. Full article

The COVID-19 pandemic prompted widespread adoption of intensified hand hygiene practices, raising concerns about their medium-term impact on the skin microbiome. This study investigates alterations in the hand microbiome of healthy adults during the pandemic compared to pre-pandemic periods in Majorca, Spain. A total of 30 volunteers (16 women, 14 men; mean age 44.1 ± 8.8 years) were sampled between 2014 and 2021. Palm swabs were collected following WHO guidelines, alongside measurements of skin pH, temperature, and handwashing frequency. Bacterial DNA was extracted and analyzed via 16S rRNA (V3-V4) metagenomic sequencing to assess microbial diversity and composition. Results revealed a significant decline in microbial diversity during the COVID-19 period, accompanied by a marked shift in the community structure. The Firmicutes phylum dominated, with Bacillales increasing from 30.7% to 84.1%, primarily driven by a surge in Staphylococcus species (e.g., S. pasteuri). Conversely, S. hominis and Actinomycetales nearly disappeared. No significant associations were observed with gender or handwashing frequency. The skin temperature increased during the pandemic, while the pH remained stable. The Staphylococcus/Bacillus ratio shifted significantly, favoring Staphylococcus dominance. These findings, derived from a geographically limited population in Majorca, Spain, demonstrate that stringent hygiene measures during COVID-19 reduced microbial diversity and restructured hand microbiome composition. The study underscores the necessity for balanced hygiene strategies that mitigate pathogen transmission while preserving beneficial microbial communities critical to skin health. Full article

Due to its high digestibility, rich nutrient profile, and potential probiotic content, goat milk is an essential nutritional resource, particularly for individuals with cow milk allergies. This review summarises the current state of microbial diversity in goat milk, emphasising the implications for quality, safety, and probiotic potential. This systematic review adhered to PRISMA guidelines, conducting a comprehensive literature search across PubMed, ScienceDirect, and Google Scholar using keywords related to microbial profiling in goat milk. The inclusion criteria targeted English-language studies from 2000 to 2025 that utilised high-throughput or next-generation sequencing methods. Out of 126 articles screened, 84 met the eligibility criteria. The extracted data focused on microbial diversity, profiling techniques, and their respective strengths and limitations in evaluating probiotic potential and spoilage risks. The review addresses the challenges linked to microbial spoilage and the composition and functional roles of microbial communities in goat milk. With species such as Bacillus and Pseudomonas playing crucial roles in fermentation and spoilage, key findings emphasise the prevalence of microbial phyla, including Proteobacteria, Firmicutes, and Actinobacteria in goat milk. The review also explores the probiotic potential of the goat milk microbiota, highlighting the health benefits associated with strains such as Lactobacillus and Bifidobacterium. Significant discoveries underline the necessity for advanced multi-omics techniques to thoroughly define microbial ecosystems and the substantial gaps in breed-specific microbiota research. Important findings illustrate the need for enhanced multi-omics techniques, given the challenges of host RNA and protein interference, low microbial biomass, and limited goat-specific reference databases, for optimising probiotic development, spoilage prevention strategies, and integrating metagenomics, metabolomics, metaproteomics, and metatranscriptomics to improve milk quality and safety as some of the future research objectives. This study emphasises the importance of understanding goat milk microbiology to advance dairy science and enhance human health. Full article

Chestnut blight, caused by Cryphonectria parasitica (Murrill) M.E. Bar, is a destructive fungal disease threatening chestnut cultivation and production. In response to the limitations of chemical control, biological control using antagonistic microbes has gained increasing attention. A rhizosphere-derived bacterium, strain D39, was isolated from healthy chestnut trees and identified as Bacillus amyloliquefaciens based on morphological characteristics and the phylogenetic analysis of 16S rRNA and gyrA genes. The antifungal activity of strain D39 against C. parasitica was evaluated using dual-culture and double-layer Oxford cup assays. The strain exhibited broad-spectrum and stable antagonistic effects and harbored five key genes associated with antimicrobial compound biosynthesis (srfAA, ituC, fenD, bmyB, and bacA), as confirmed by PCR. A 145 kDa extracellular protein with strong antifungal activity was extracted and purified by ammonium sulfate precipitation, DEAE ion-exchange chromatography, and Sephadex G-75 gel filtration. LC-MS analysis identified the protein as a serine peptidase belonging to the S8 family, and its structure was predicted using multiple bioinformatic tools. In pot experiments, treatment with the strain D39 significantly reduced disease severity, achieving control efficiencies of 66.07% and 70.89% at 10 and 20 days post-inoculation, respectively. These results demonstrate that B. amyloliquefaciens D39 has strong potential as a biocontrol agent against chestnut blight, offering an effective and environmentally friendly alternative for disease management. Full article

The Rio Bravo/Grande River is a binational water resource between Mexico and the United States and supports diverse anthropogenic activities. However, limited studies on its microbiological composition focus on molecular techniques. Therefore, the aim of this study was to characterize the bacteriome and identify potentially pathogenic bacteria in surface waters of the Rio Bravo/Grande in northeastern Tamaulipas, Mexico, using the 16S rRNA gene metabarcoding technique. Surface water samples were collected from the localities of Diaz Ordaz, Reynosa, and Matamoros between 2016 and 2017. DNA extraction and sequencing were performed, focusing on the V3–V4 region of the 16S rRNA gene. A taxonomic analysis revealed the presence of 13 bacterial phyla, with Proteobacteria (40%), Firmicutes (28%), and Actinobacteria (10.2%) being the most abundant. At the genus level, Bacillus, Pseudomonas, and Acinetobacter were predominant, while 16 potentially pathogenic species, including Acinetobacter baumannii and Vibrio vulnificus, were identified. Alpha and beta diversity analyses highlighted significant differences in the bacterial diversity among the sampling sites, indicating that the river has some capacity to recover from anthropogenic and environmental disturbances. This study underscores the need for the continuous monitoring of the Rio Bravo/Grande to protect public health and maintain the water quality in the face of increasing anthropogenic pressures. Full article

Crab roe sauce (CRS) is prone to spoilage due to microbial contamination. Therefore, this study aimed to investigate the inhibitory effects and mechanisms of a mixed bio-preservative (0.025% ε-polylysine hydrochloride (ε-PL) + 0.01% nisin (NS) + 0.01% tea polyphenols (TPs)) on the specific spoilage bacteria (SSB) in CRS. First, the SSB in CRS were isolated and identified by 16S rRNA sequencing. Two isolates were selected as representative strains based on their enzymatic spoilage potential and spoilage capability in CRS. By comparing the inhibition zones, ε-PL, NS, and TPs were selected from five conventional bio-preservatives (ε-PL, NS, TPs, grape seed extract (GSE), and rosemary extract (RE)) to prepare the mixed bio-preservative. The results showed that the minimum inhibitory concentration (MIC) of the mixed bio-preservative against Bacillus pumilus and Bacillus subtilis was 56.3 µg/mL. The growth curves and cell viability tests revealed that the mixed bio-preservative reduced the viability of both strains. The conductivity, alkaline phosphatase activity, and nucleic acid and soluble protein leakage indicated that the mixed bio-preservative disrupted the integrity of the cell walls and membranes of the two isolates in a concentration-dependent manner. Scanning electron microscopy further confirmed the damage to the cell membranes of the two isolates by the mixed bio-preservative. Overall, the mixed bio-preservative exhibited excellently inhibitory effects on the SSB and could be a promising method for the preservation of CRS. Full article

Ticks are important vectors of pathogenic bacteria that cause diseases in both humans and animals. Analysis of tick microbiota via massive sequencing allows rapid and comprehensive identification of almost all bacteria inhabiting ticks. This has improved the detection of emerging pathogens and has helped define their relationship with public health. In Mexico, the brown dog tick (Rhipicephalus sanguineus sensu lato) is a public health problem, especially in northeast Durango. In the present study, the bacterial microbiota of this tick was determined using third-generation massive sequencing (PacBio, V1–V9 region of the 16S rRNA gene); bacteria with pathogenic potential that are transmitted by salivation and those that can be transmitted by accidental regurgitation of the parasite were also identified. In 2024, 60 dogs were searched for unfed ticks; then, 15 groups of female ticks and 15 groups of male ticks were formed, with each group consisting of 30 individuals. DNA was extracted from each tick pool, and the complete 16S rRNA gene was amplified (PacBio). Bioinformatics analysis was performed in QIIME2 (Quantitative Insights into Microbial Ecology) to obtain amplicon sequence variants (ASVs). Alpha and beta diversity metrics, as well as statistical analyses, were performed to test for differences between the microbiota of females and males. The bacterial taxa were classified into 21 phyla, 24 classes, 81 orders, 137 families, 339 genera, and 565 species. The male microbiota presented a significantly greater number of ASVs and a greater phylogenetic diversity index (FaithPD). Additionally, the unweighted UniFrac metric was significantly different between the sexes. The endosymbiont Coxiella mudrowiae was significantly more abundant in females, and Ehrlichia canis was more abundant in males. The pathogens E. canis and Anaplasma platys (transmitted by salivation) were detected, as well as 75 species of potential pathogens recorded in this tick that could enter the host in case of accidental regurgitation of the parasite (e.g., Staphylococcus, Streptococcus, Acinetobacter, Corynebacterium, Proteus, Clostridum, Klebsiella, Bacillus, Escherichia, Fusobacterium, and Pasteurella). It is necessary to continue analyzing the microbiota of ticks through massive sequencing for the benefit of public health and to establish new alternatives for controlling these parasites. Full article

Background/Objectives: Research-engaged academic institutions offer the opportunity to couple undergraduate education/citizen science projects with antimicrobial biodiscovery research. Several initiatives reflecting this ethos have been reported internationally (e.g., Small World, Tiny Earth, MicroMundo, Antibiotics Unearthed). These programs target soil habitats due to their high microbial diversity and promote initial screening with non-selective, nutrient media such as tryptic soy agar (TSA). However, evaluation of published outputs to date indicates that isolate recovery on TSA is consistently dominated by the genera Bacillus, Pseudomonas, and Paenibacillus. In this study, we evaluated the potential of soil extract agar to enhance soil isolate diversity and antibiosis induction outcomes in our undergraduate Antibiotics Unearthed research program. Methods: We comparatively screened 229 isolates from woodland and garden soil samples on both tryptic soy agar (TSA) and soil extract agar (SEA) for antimicrobial activity against a panel of clinically relevant microbial pathogens. Results: On one or both media, 15 isolates were found to produce zones of clearing against respective pathogens. 16S rRNA gene sequencing linked the isolates with three genera: Streptomyces (7), Paenibacillus (6), and Pseudomonas (2). Six of the Streptomyces isolates and one Pseudomonas demonstrated antimicrobial activity when screened on SEA, with no activity on TSA. Furthermore, incorporation of the known secondary metabolite inducer N acetyl-glucosamine (20 mM) into SEA media altered the pathogen inhibition profiles of 14 isolates and resulted in broad-spectrum activity of one Streptomyces isolate, not observed on SEA alone. In conclusion, SEA was found to expand the diversity of culturable isolates from soil and specifically enhanced the recovery of members of the genus Streptomyces. SEA was also found to be a superior media for antibiosis induction among Streptomyces isolates when compared to TSA. It was noted that Paenibacillus isolates’ antibiosis induction demonstrated a strain-specific response with respect to the growth media used. Conclusions: The authors propose SEA inclusion of in soil screening protocols as a cost-effective, complementary strategy to greatly enhance outcomes in undergraduate/citizen science-engaged antimicrobial biodiscovery initiatives. Full article

Vanilla planifolia is grown as a high-value orchid spice for its odor and savor attributes that increase due to the curing process associated with microbial colonization. This tends to influence the aromatic properties of vanilla. Hence, 11 Bacillus sp. strains were isolated from V. planifolia and identified with 16S rRNA gene sequencing. The liquid culture (1 mL of 10⁷ CFU mL⁻¹) of selected Bacillus vallismortis NR_104873.1:11-1518, Bacillus velezensis ZN-S10, and Bacillus tropicus KhEp-2 effectively fermented green-blanched vanilla pods kept at 10 °C during the sweating stage. GC-MS analysis showed that the methanol extract of non-coated, and B. vallismortis treated vanilla detected three (3) volatile compounds, whereas seven (7) components were obtained in B. tropicus and B. velezensis treatment. 4H-pyran-4-one, 2,3-dihydro-3,5-dihydroxy-6-methyl was found in B. velezensis ZN-S10, B. tropicus KhEp-2, and B. vallismortis while it was not present in the control samples. This ketone compound suggested a Maillard reaction resulting in brown-increased aroma pods. Linoleic acid and Hexadecanoic acid ethyl esters were detected only in ZN-S10 strain-coated vanilla. A novel 3-Deoxy-d-mannoic lactone was detected only in B. vallismortis-treated vanilla characterized as a new compound in V. planifolia which suggested that the new compound can be altered with the coating of bacteria in vanilla during fermentation. Thus, the Bacillus strains improved the volatile profile and exhibited a new aroma and flavor profile of vanilla owing to bacteria fermentation during the curing process. Full article

In the current investigation, the antibacterial activity of zinc oxide nanoparticles (ZnO-NPs) formed by an aqueous extract of Psidium guajava leaves against foodborne pathogenic bacterial strains was investigated. To achieve this goal, 33 bacterial isolates were obtained from spoiled fruits. Among these isolates, 79% showed cellulase activity, 82% showed amylase activity, 81% exhibited xylanase potential, and 65% exhibited lipase activity. Moreover, 12 isolates showed complete hemolysis (β-hemolysis). The identification of these isolates was done using sequencing and amplification of 16s rRNA as Staphylococcus aureus (two strains), Pseudomonas syringae (one strain), E. coli (two strains), Salmonella typhimurium (two strains), Listeria monocytogenes (one isolate), Bacillus cereus (two isolates), and Bacillus subtilis (two isolates). The formed ZnO-NPs by aqueous Psidium guajava leaf extract were characterized using UV, FT-IR, TEM, EDX, XRD, DLS, and Zeta potential. The data revealed the successful formation of a spherical shape, crystallographic structure, and well-arranged ZnO-NPs. FT-IR showed the effect of different functional groups in the plant extract in the formation of ZnO-NPs through reducing, capping, and stabilizing of end products. Moreover, EDX analysis showed that the Zn ion occupied the main component of the produced NPs. Interestingly, the obtained bacterial strains showed varied sensitivity toward green-synthesized ZnO-NPs. The growth inhibition of foodborne pathogenic strains by ZnO-NPs was concentration dependent, forming a zone of inhibition in the range of 20–23 mm at a concentration of 200 µg mL⁻¹, which decreased to 15–18 mm at 100 µg mL⁻¹. Moreover, the values of MIC were 25 and 50 µg mL⁻¹ based on the bacterial strain. Overall, the green-synthesized ZnO-NPs can be a useful approach for inhibiting the growth of spoilage bacterial strains that destroy fruits and hence reduce the harmful effects of traditional treatment methods on the environment and human health. Full article

In this study, a microbial strain was isolated from humus soil to ferment Sophora flavescens and Eleutherococcus sessiliflorus extracts. The isolated microbial was identified as the Bacillus genus by 16S rRNA sequence analysis. The fermented plant extracts exhibited antifungal effects against four types plant pathogen, P. carotorum, B. cinerea, C. fructicola Sau-3, and C. gloeosporioides, according to incubation time. In particular, the fermented plant extracts showed the most activity for Colletotrichum genus in inhibiting mycelium growth. Metabolite changes in fermented S. flavescens and E. sessiliflorus extracts were confirmed through LC-Q-TOF/MS. Flavonoid and peptide derivatives were improved in fermented S. flavescens and E. sessiliflorus extracts compared to their unfermented counterparts. This study suggested that isolated Bacillus microbial fermentation could be a valuable tool in improving the bioactivity of S. flavescens and E. sessiliflorus extracts, with the potential to form more environmentally friendly antifungal agents. Full article

Fire blight, a devastating disease caused by Erwinia amylovora, poses a significant threat to pear and apple trees in Xinjiang province, China. In an effort to combat this pathogen, we isolated 10 bacteria from various components of apple and crabapple trees and conducted screenings to assess their ability to inhibit E. amylovora in vitro. Through biochemical tests and partial 16S rRNA gene sequencing, we identified two promising strains, Priestia megaterium strain H1 and Bacillus subtilis strain I2. These strains were then evaluated for their efficacy in biocontrol under controlled laboratory conditions, focusing on immature fruits and leaves. Remarkably, all selected antagonists exhibited the capability to reduce the severity of the disease on both fruit and leaves. P. megaterium strain H1 and B. subtilis strain I2 exhibited significant reductions in disease incidence on both immature fruits and leaves compared to the control. Specifically, on immature fruits, they achieved reductions of 53.39% and 44.76%, respectively, while on leaves, they demonstrated reductions of 59.55% and 55.53%, respectively. Furthermore, during the study, we detected the presence of lipopeptides, including surfactin, iturins, bacillomycin D, and fengycins, in the methanol extract obtained from these two antagonistic bacteria using thin-layer chromatography (TLC). Based on the results obtained, B. subtilis strain I2 and P. megaterium strain H1 exhibit considerable potential for controlling fire blight. However, further evaluation of their efficacy under natural field conditions is essential to validate their practicality as a biocontrol method. Full article