International Microbiology最新文献

The main objective of this study was to assess cellulolytic probiotic strains from traditional fermented beverages such as palm wine in order to supplement the animal feed and strengthen the gut health of the animal for better digestibility and absorption. In the present study, different types of microbes were isolated from traditionally prepared palm wine and analyzed for their probiotic nature. For any microbe to be probiotic in nature, it has to sustain the harsh conditions of the human gastrointestinal tract such as acid tolerance, bile tolerance at the lower range of pH, and other properties like auto aggregation test, cell surface hydrophobicity test with non-polar hydrocarbons for evaluating its capabilities to adhere to the intestinal cells and antimicrobial nature against pathogens. Bacillus mycoides strain PR04 and Bacillus subtilis strain PR21 were found to be resistant to acid and bile in simulated artificial gastrointestinal tract model, found to be than 55% hydrophobic with xylene and n-hexadecane and also showed antimicrobial activity greater towards pathogenic strains like Pseudomonas aeruginosa, Staphylococcus aureus, Candida albicans, and Salmonella typhimurium respectively. The cellulolytic activity of the isolates PR04 and PR21 was evaluated in (0.2–2) % CMC (carboxymethyl cellulose) plate. Bacillus mycoides PR04 and Bacillus subtilis PR21 could degrade carboxymethyl cellulose, filter paper, and sugarcane bagasse. The degradation of sugarcane bagasse was confirmed by Scanning electron microscopy and filter paper degradation after 4 days of incubation at 37 °C. Cellulase gene of the identified Bacillus sp. strains was amplified by primers CF5′-ACAGGATCCGATGAAAACGGTCAATTTCTATTTT-3′ and CR5′-ACTCTCGAGATTGGGTTCTGTTCCCAAT-3′. This study proposes potential probiotic Bacillus mycoides PR04 (Accession no. OR625070) and Bacillus subtilis PR21 (Accession no. OR625072) in the application as an animal feed additive to assist in its digestibility and encourage the gut health.

The broad-spectrum antibacterial capabilities of fatty acids (FAs) and their reduced propensity to promote resistance have rendered as a promising substitute for conventional antibiotics. The structural significance of fatty acid production with the other lipids is a major energy source, and signal transduction has drawn a great deal of research attention to these biomolecules. Saturated and monounsaturated fatty acids reduce virulence by preventing harmful opportunistic bacteria like Pseudomonas aeruginosa and Chromobacterium violaceum from activating their quorum sensing (QS) systems. In this finding, the fatty acids capric acid, caprylic acid, and monoelaidin were selected to evaluate their anti-QS activity against the C. violaceum and P. aeruginosa. At the minimum inhibitory concentration (MIC) and sub-MIC concentration of the three fatty acids, the virulence factor production of both the bacteria was quantified. The virulence factors like EPS, biofilm quantification and visualization, and motility assays were inhibited in the dose-dependent manner (MIC and sub-MIC) for both the organisms whereas this pattern was followed in the pyocyanin, pyoverdine, rhamnolipid, protease of P. aeruginosa and the violacein, and chitinase of C. violaceum. In all these biochemical assays, the capric acid could effectively reduce the production and further validated at gene expression level by RT-qPCR. The study on the gene expression for all these virulence factors reveals that the capric acid inhibited the growth of both the organisms in a higher fold than the caprylic and monoelaidin. The in silico approach of structural validation for the binding of ligands with the proteins in the QS circuit was studied by molecular docking in Schrodinger software. The Las I and Las R in P. aeruginosa and the CviR of C. violaceum protein structures were docked with the selected three fatty acids. The capric acid binds to the pocket with the highest binding score of all the proteins than the caprylic and monoelaidin fatty acids. Thus, capric acid proves to be the therapeutic biomolecule for the anti-QS activity of opportunistic bacteria.

Graphical Abstract

Background

Coagulase-negative staphylococci (CoNS) are evolving as major reservoirs and vectors of unusual and critical antimicrobial resistance (AMR) mechanisms.

Materials and methods

In this study, the genomic characterization of 26 multidrug-resistant (MDR)-CoNS (S. borealis, S. saprophyticus, S. sciuri, S. hominis, S. epidermidis, S. pasteuri, S. hyicus, S. simulans, S. haemolyticus, and S. arlettae) previously obtained from the nasal cavity of healthy nestling storks, humans who had no contact with animals, pigs, and pig farmers, as well as dogs and dog owners from Spain was performed. High-quality draft genomes obtained by Illumina sequencing technology were used to determine their resistome, virulome, mobile genetic elements, and CRISPR-Cas types.

The relatedness of three CoNS species with publicly available genomes was assessed by core-genome single nucleotide polymorphisms (SNPs).

Results

AMR genes to all classes of antibiotics in staphylococci were detected including unusual ones (mecC, ermT, and cfr), of which their corresponding genetic organizations were analyzed. About 96.1% of the MDR-CoNS strains harbored diverse adherence or immune evasion genes. Remarkably, one enterotoxin-C and -L-carrying S. epidermidis-ST595 strain from a nestling stork was detected. Moreover, various plasmid bound-biocide resistance genes (qacACGJ) were identified in 34.6% of the MDR-CoNS. Two genes that encode for cadmium and zinc resistance (cadD, czrC) were found, of which czrC predominated (42.3%). Complete CRISPR-Cas system was detected in 19.2% of the CoNS strains, of which cas-1, -2, and -9 predominated, especially in 75% of the S. borealis strains. The phylogenetic analysis identified clusters of related S. epidermidis lineages with those of other countries (SNP < 100). Also, highly related S. borealis isolates (SNP < 10) from pigs was confirmed for the first time in Spain.

Conclusion

These findings showed that various ecological niches harbor CoNS that presented MDR phenotypes mediated by multiple AMR genes carried by mobile genetic elements with relatively low frequency of intact CRISPR-Cas systems. Furthermore, the transmission of some CoNS species in humans and animals is strongly suggested.

Free-living amoebae infections are on the rise while the prognosis remains poor. Current therapies are ineffective, and there is a need for novel effective drugs which can target Naegleria, Balamuthia, and Acanthamoeba species. In this study, we determined the effects of a nano-formulation based on flavonoid patuletin-loaded gallic acid functionalized zinc oxide nanoparticles (PA-GA-ZnO) against Acanthamoeba, Balamuthia, and Naegleria trophozoites. Characterization of the nano-formulation was accomplished utilizing analytical tools, namely Fourier-transform infrared spectroscopy, drug entrapment efficiency, polydispersity index, dimensions, and surface morphologies. Anti-amoebic effects were investigated using amoebicidal assay, cytopathogenicity assay, and cytotoxicity of the nano-formulation on human cells. The findings revealed that nano-formulation (PA-GA-ZnO) displayed significant anti-amoebic properties and augmented effects of patuletin alone against all three brain-eating amoebae. When tested alone, patuletin nano-formulations showed minimal toxicity effects against human cells. In summary, the nano-formulations evaluated herein depicts efficacy versus Acanthamoeba, Balamuthia, and Naegleria. Nonetheless, future studies are needed to comprehend the molecular mechanisms of patuletin nano-formulations versus free-living amoebae pathogens, in addition to animal studies to determine their potential value for clinical applications.

Obligate endosymbiont bacteria associated with insects are naturally providing their hosts with essential nutrients such as vitamins and amino acids and biological services including protection from pathogens. In this study, we aimed to investigate the presence of Wolbachia infection among males and females of the parasitic apricot seed wasp (ASW) Eurytoma samsonowi Vassiliev (Vassiliev Petrograd 11: 1-15, 1915) (Hymenoptera: Eurytomidae), a very harmful pest of apricot (Prunus armeniaca), in the oasis of Gafsa, Southern-West of Tunisia. The detection of Wolbachia infection was assessed based on the amplification of the Wolbachia surface protein (wsp) gene and a multilocus sequence typing (MLST) as a universal genotyping tool for Wolbachia involving the analyses of genes gatB, coxA, hcpA, fbpA, and ftsz. Confirming the screening results, Wolbachia was detected in the natural apricot wasp for the first time, with a significant difference between males (5%) and females (59%) based on wsp gene. All Wolbachia strains identified in E. samsonowi were clustered among supergroups B of Wolbachia.

Thermophilic bacteria able to produce exopolysaccharides (EPSs) have become attractive in recent years. EPSs synthesized by thermophiles are worth investigating due to their unexplored structural and biological properties. In this study, EPSs from thermophilic, Gram-positive bacterial isolates were purified and tested for their biological activities. A total of one hundred seven thermophilic bacteria were screened for their ability to produce exopolysaccharides. Nine isolates belonging to Geobacillus, Parageobacillus, Aeribacillus, and Anoxybacillus genera with highest EPS production were chosen, and purified EPSs (20, 61, 74, 76, 78, 89, 106, 134, and 261) were used for biological activity studies. EPS yields of selected thermophilic bacteria ranged between 117 and 419 mg/L. Among the tested EPSs, 61, 106, and 261 showed antibacterial effect against E. faecalis JH2-2 at a final concentration of 1.5 mg/mL. EPS samples had significant antioxidant capacity, especially EPS 134, with the highest DPPH radical scavenging activity of 100% at a concentration of 5 mg/mL and the strongest reducing power. EPS 20 showed the highest lipid peroxidation inhibition effect at a rate of 31%. EPSs displayed weak alpha-amylase inhibition activity when compared with standard acarbose. The prebiotic indices of EPSs 20, 61, 76, 89, 134, and 261 were found to be higher than that of inulin, a representative prebiotic carbohydrate for all tested lactic acid bacteria in the study. All examined EPSs inhibited the biofilms formed by various bacteria depending on the test strain. Results indicated that thermophilic EPSs had remarkable antioxidant, prebiotic, and antibiofilm activities. Therefore, EPSs characterized in this study may have technological applications in health and food fields.

Chilli leaf curl disease (ChiLCD), which is a significant problem in chilli cultivation, is caused by begomoviruses that are transmitted by the whitefly Bemisia tabaci. This disease leads to severe impacts on crop yields. To determine the incidence of begomovirus in the chilli crop, infected chilli leaf samples exhibiting symptoms such as curling, yellowing, reduced leaf size, and overall stunted growth were collected from various districts of Tamil Nadu, namely, Coimbatore, Dharmapuri, Kancheepuram, Karur, Salem, Krishnagiri, Thoothukudi, Thiruvallur, Tiruchirappalli, Virudhunagar, Tiruvannamalai, Tenkasi, and Vellore, during the years 2018-2022. To determine the complete genome sequence of the begomoviruses, the rolling circle amplification (RCA) method was used to clone and sequence the begomovirus genomes from the chilli samples collected from various districts of Tamil Nadu. Here we characterized 17 DNA A genome sequences and 12 betasatellite sequences. BLAST results of the DNA A genome sequences revealed nucleotide identities ranging from 94.2 to 99.7% with five distinct begomovirus species of chilli, namely, chilli leaf curl Salem virus (HM007119), chilli leaf curl virus Bhavanisagar (NC_055130), chilli leaf curl Ahmedabad virus (MW795666), chilli leaf curl virus (NC_055131), and chilli leaf curl Sri Lanka virus (JN555600). BLAST results of the betasatellite sequences showed nucleotide identities of 96 to 98.8% with the tomato leaf curl Bangladesh betasatellite (MZ151286). In the present study, five distinct begomovirus species and one associated betasatellite were found to infect chilli crops in Tamil Nadu. This finding indicates a changing pattern of begomovirus occurrence in the different districts of Tamil Nadu. This study highlights the prevalence of chilli-infecting begomoviruses in the major chilli growing districts of Tamil Nadu, the identification of begomovirus species, and the significance of understanding and managing these viruses to safeguard chilli cultivation in the region.

This study investigated the potential applications of Enterococcus hirae MLG3-25-1 exopolysaccharides (EPS), with a focus on their isolation, identification, production, and functional characteristics. After the bacterial strain was cultured in De Man-Rogosa-Sharpe (MRS) medium containing 1% glucose at 37 °C, the EPS was refined, and the highest yield of 0.85 mg/mL was achieved at the 24-h incubation period. Enterococcus hirae MLG3-25-1 was found to be able to produce EPS. The study explored the microstructure of the EPS, which resembles polysaccharide sheets with smooth surfaces, through scanning electron microscope (SEM) analysis. Through Fourier transform infrared spectroscopy (FT-IR) and nuclear magnetic resonance (NMR) analysis, the chemical composition, aligning with glycosidic bond characteristics, has been deciphered. Furthermore, the antimicrobial and antibiofilm activities against pathogenic bacteria, particularly Bacillus sp., demonstrated potential applications in combating antibiotic resistance. The EPS exhibited notable antioxidant activity (89.36% DPPH scavenging), along with high water-holding capacity (575%), emulsifying activity, and flocculation activity, suggesting its potential as a stabilizing agent in the food industry. Overall, this study provides a comprehensive characterization of Enterococcus hirae MLG3-25-1 EPS, emphasizing its diverse applications in antimicrobial, antioxidant, and food-related industries. These findings lay the groundwork for further exploration and utilization of this EPS in various sectors.

In the present study, we assessed the antioxidant and antileishmanial potential from fresh leaves of Ballota (B.) hirsuta essential oil (EO). The GC-MS analysis of B. hirsuta EO revealed that spathulenol and germacrene D were the main components accounting for 26.03% and 19.64% of the total EO, respectively. B. hirsuta EO possesses moderate antioxidant activity, both in neutralizing DPPH radicals and in inhibiting β-carotene bleaching. In addition, it exhibits both high antileishmanial activity and selectivity towards the promastigote and amastigote forms. Specifically, B. hirsuta EO showed an IC₅₀ value of 20.78 µg/mL and 23.62 µg/mL, against the promastigote and amastigote forms of L. infantum, respectively. It also demonstrated an IC₅₀ value of 22.39 and 25.76 µg/mL, against the promastigote and amastigote forms of L. major, respectively. However, it exhibited moderate cytotoxicity, with a selectivity index below 10. The investigation of the molecular mechanism of action revealed that B. hirsuta EO inhibited only the sterol pathway, including CYP51 gene expression. Additionally, in silico analysis indicated that the main compounds of B. hirsuta EO, germacrene and spathulenol, exhibited excellent affinity energy against Leishmania enzymes trypanothione reductase (TryR) and trypanothione synthase (TryS). This denotes the potential of these compounds as promising agents to control leishmaniasis.

The rise of antibiotic resistance in existing pathogens has been identified as a major threat to global healthcare in the twenty-first century. This resistance has consequences such as increased cost and prolonged hospital stays, treatment failure, and ultimately increased risk of patient mortality. It is therefore imperative to develop strategies to combat drug resistance. Combined treatment of common antibiotics and natural compounds is one of the most effective methods against resistant bacterial infections. Gallic acid (GA) is a natural secondary metabolite abundantly found in plants and has significant medicinal effects in various aspects of health. In this research, the antibacterial effects of azithromycin (AZM) and GA alone and in combination with each other were investigated on planktonic and biofilm forms of methicillin-sensitive Staphylococcus aureus (MSSA), methicillin-resistant Staphylococcus aureus (MRSA), and Pseudomonas aeruginosa (P. aeruginosa). The results showed that the combination of AZM/GA had an additive effect against MSSA and P. aeruginosa and a synergistic effect against MRSA. In addition, combining these two agents significantly reduced the minimum biofilm inhibitory concentration (MBIC) of AZM and GA in the MRSA strain. Finally, the level of ROS generation in the effect of AZM plus GA was evaluated in the bacteria. Among the studied strains, ROS production was significantly increased in combination treatment compared to AZM alone in MRSA. The results show that the combination of AZM and GA has a significant effect against MRSA and can be considered as an effective treatment option.