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In numerous countries, the utilization of plants for both nutritional and therapeutic purposes is a common practice. However, the inadvertent use of these plants can pose risks due to their active molecules or microbiota. The traditional use of Herniaria glabra L. (H. glabra) plant in treating various diseases is well-known; however, its application in yogurt production raises concerns. In this study, Bacillus pumilus isolated from H. glabra was identified through 16s rRNA sequencing and MALDI-TOF MS (Matriks Assisted Lazer Desorption Ionization Time of Flight Massspectrometry). The bacterium's resistance under simulated gastrointestinal tract (GIT) conditions was assessed, followed by investigations into its aggregation ability, antibiotic resistance, hemolytic activity, and antagonistic potential through in vitro tests. The study revealed that B. pumilus exhibited 100% resistance to GIT conditions. Notably, the bacterium demonstrated strong autoaggregation (34.48%) and coaggregation abilities (49.82% for Escherichia coli, 49.13% for Listeria monocytogenes), signifying a potent aggregative potential. Sensitivity to most tested antibiotics was observed, while no antagonistic activity against tested bacteria was evident. Furthermore, the bacterium exhibited β-hemolytic activity, indicative of potential virulence. The findings suggest that this resistant yet virulent bacterium, with its hemolytic activity, could disrupt the GIT balance, posing serious health risks. The study underscores the need for caution and awareness regarding the potential dangers posed by bacteria in plant microbiota in herbal therapies. [ABSTRACT FROM AUTHOR]

Polygalacturonases (PG) are the enzymes that cause depolymerization of pectin. In current study, bacterial strain was isolated from rotten sample and then purified. It was screened for endo-polygalacturonase activity using PSAM media. It had shown positive response for endo-PG activity. Bacterial DNA was isolated and 16 S rRNA gene amplification was done using universal primer pair of 8 F and 1492 R. Bacterial strain was also amplified by 16 S rRNA gene for sequencing. BLAST of gene sequence on NCBI database had shown that bacterial isolate was identified as Bacillus pumilus. Endo-polygalacturonase enzyme was produced by submerged fermentation to optimize culture conditions by RSM in JMP-12 software. The optimized parameters had shown maximum endo-polygalacturonase activity of 153.2 U/mL/min after using 3 g of orange peels as substrate, 3 mL of inoculum size, 6.5 pH buffer, 40°C temperature and 3 days of incubation. After optimizing fermentation conditions, endo-polygalacturonase was precipitated using 70 % concentration of ammonium sulfate. This partially precipitated sample was dialyzed with dialysis tube and used to find activity of endo-polygalacturonase (1620.6 U/mL/min). This sample was applied to gel filtration chromatography for further purification. Endo-polygalacturonase activity increased many times 2231.44 U/mL/min after gel filtration. The Molecular weight of endo-polygalacturonase was 46 kDa after SDS PAGE characterization. High Vmax value and alkaliphilic nature of endo-polygalacturonase will make it good candidate for food and feed industry near future. [Display omitted] • A bacterial strain was isolated from rotten sample. • Bacillus pumilus was identified through 16 S rRNA gene sequencing. • Maximum enzyme activity of 153.2 U/mL/min was achieved under optimized conditions. • The characterized endo-polygalacturonase has great potential in industrial applications. • Polygalacturonase had shown maximum activity after column chromatography. [ABSTRACT FROM AUTHOR]

The efficacy of ultra-high-pressure homogenisation (UHPH) in inactivating Bacillus pumilus ATCC 27142 and Bacillus subtilis ATCC 6633 spores suspended in sheep and cow milk was investigated. The UHPH treatment was conducted at 200 and 250 MPa with an inlet temperature of 85 °C, resulting in homogenising valve temperatures of 117 °C and 127 °C, respectively. To isolate the role of temperature and pressure in the inactivation of bacterial spores, the UHPH treatment was repeated at 250 MPa with a lower inlet temperature of 70 °C that resulted in a valve temperature of 117 °C. Increasing the pressure and valve temperature resulted in increased inactivation. At 250 MPa with a valve temperature of 127 °C, greater than 5 log CFU/mL reduction was achieved in B. pumilus and B. subtilis spores in both milk types. Reductions of 0.61 ± 0.03 log CFU/mL and 0.62 ± 0.09 log CFU/mL in B. pumilus spores and 1.18 ± 0.04 log CFU/mL and 1.30 ± 0.07 log CFU/mL in B. subtilis spores were obtained at 250 MPa with a valve temperature of 117 °C in sheep and cow milk, respectively. The spore inactivation was influenced by both the pressure and temperature, suggesting a synergistic effect, with the latter playing a critical role in the lethality of the treatment. No significant differences in the inactivation of either strain was observed between sheep and cow milk. [ABSTRACT FROM AUTHOR]

Stem nematode (Ditylenchus destructor Thorne) is considered one of the most economically devastating species affecting sweet potato production. Biocontrol offers a sustainable strategy for nematode control. This study conducted a pot experiment to evaluate the biocontrol efficacy of Paecilomyces lilacinus CS-Z and Bacillus pumilus Y-26 against the stem nematode, as well as to examine their influence on the bacterial communities in the sweet potato rhizosphere. The findings indicated that B.pumilus Y-26 and P.lilacinus CS-Z exhibited respective suppression rates of 82.9% and 85.1% against the stem nematode, while also stimulating sweet potato plant growth. Both high-throughput sequencing and Biolog analysis revealed distinct impacts of the treatments on the bacterial communities. At the phylum level, B.pumilus Y-26 enhanced the abundance of Actinobacteria but reduced the abundance of Cyanobacteria, with P.lilacinus CS-Z exhibiting similar effects. Additionally, the treatment with B.pumilus Y-26 resulted in increased abundances of Crossiella, Gaiella, Bacillus, and Streptomyces at the genus level, while the treatment with P.lilacinus CS-Z showed increased abundances of Crossiella and Streptomyces. In contrast, the abundance of Pseudarthrobacter was reduced in the treatment with B.pumilus Y-26. Conversely, the application of the nematicide fosthiazate exhibited minor influence on the bacterial community. The findings indicated that the application of P.lilacinus CS-Z and B.pumilus Y-26 led to an increase in the relative abundances of beneficial microorganisms, including Gaiella, Bacillus, and Streptomyces, in the rhizosphere soil. In conclusion, P.lilacinus CS-Z and B.pumilus Y-26 demonstrated their potential as environmentally friendly biocontrol agents for managing stem nematode disease of sweet potato. [ABSTRACT FROM AUTHOR]

Oak decline is a major concern in Iran. A literature search with the terms 'Iran' AND 'oak' OR 'Quercus' from 1963 through 2023 resulted in 635 articles with over 200 dealing specifically with oak decline. In Iran, oak decline is most severe in the Zagros forests, followed by the Arasbaran and Hyrcanian forests. Of Iran's 11 native oak species, Quercus brantii is the most affected. Several factors have been identified as contributing to oak decline in Iran. Some economic and social factors include high rural unemployment, livestock overgrazing and uncontrolled cutting of live tree branches with the wood used for firewood and foliage used as food for farm animals. These pruning wounds can serve as entry points for pathogens and attract insects. Important abiotic factors included extended drought, increasing air temperature and wind speed, and increased levels of airborne dust and heavy metals. Important pathogens identified in the early 2000s included Armillaria mellea and charcoal rot fungus (Biscogniauxia mediterranea). More recently, many other fungal pathogens (e.g., Discula quercina, Neoscytalidium dimidiatum, Obolarina persica, Paecilomyces formosus and Phaeoacremonium tuscanicum) and bacterial pathogens (e.g., Bacillus pumilus, Brenneria goodwinii, Gibbsiella quercinecans, Rahnella victoriana and Stenotrophomonas maltophilia) were found in association with declining oaks. Among the insects, severe oak defoliation has resulted from various Lepidoptera in the families Erebidae, Geometridae and Tortricidae. Similarly, several bark‐ and wood‐boring beetles in the families Buprestidae and Cerambycidae were reported from declining oaks, with the buprestid Agrilus hastulifer being one of the most common. It is hoped that this review will improve our understanding of the many factors involved in oak decline in Iran and promote the development of management strategies. [ABSTRACT FROM AUTHOR]

One of the most important phytosanitary diseases affecting the production and marketing of the 'Hass' avocado fruit is anthracnose, mainly caused by the fungus Colletotrichum gloeosporioides. This disease is controlled with synthetic fungicides, which can cause harm to humans and the environment. Due to this, safer alternatives have been sought, such as biological control. The objective of this study was to isolate and characterize Bacillus spp. strains that biocontrol C. gloeosporioides in 'Hass' avocado fruits. The isolation, purification, and molecular identification of bacteria with biocontroller capacity was carried out. In vitro and in vivo tests were carried out against the pathogen C. gloeosporioides. Furthermore, the percentage inhibition of spore germination, enzymatic profile, and nutrient assimilation were determined. Forty-three native bacterial strains were isolated from the 'Hass' avocado fruit. The strains that showed the greatest antagonistic capacity in vivo against C. gloeosporioides were Bacillus pumilus AB31, B. thuringiensis AB30, B. thuringiensis AB7, and B. thuringiensis AB21. These strains could produce extracellular enzymes, such as phosphatases, proteases, and β-glucosidase, as well as the assimilation of D-ribose and trehalose. The above favors their antagonistic activity. Four strains of the genus Bacillus (one B. pumilus and three B. thuringiensis) can control C. gloeosporioides in 'Hass' avocado. [ABSTRACT FROM AUTHOR]

Human papillomavirus type 16 (HPV16) belongs to viruses of the high-risk type and is associated by overexpression of E6 and E7 oncoproteins, which determine the oncogenic properties of the virus, such as immortalization and malignant transformation of proliferating epithelial cells. The biogenesis of redox-sensitive proteins E6 and E7 at the early stages of viral infection leads to blocking of the cell antioxidant defense system and ubiquintin-dependent degradation of p53 and Rb tumor suppressors. Maintaining high rates of tumor cell proliferation contributes to an increase in the level of reactive oxygen species (ROS) and a shift in the redox balance towards oxidative processes. Reduced glutathione (GSH) provides antioxidant protection to tumor cells through S-glutathionylation of thiol groups of redox-sensitive proteins, which leads to the appearance of multidrug-resistant forms of cancer. In this regard, drugs restoring redox balance and increasing susceptibility to antitumor therapy are of particular importance. We have established that, Bacillus pumilus RNase (binase) modulates the redox-dependent regulatory mechanisms that ensure tumor cell resistance to apoptosis in HPV-16-positive SiHa cells of cervical squamous cell carcinoma,. Binase in nontoxic concentrations initiates a number of pre-apoptogenic changes, i.e., decreases ROS and reduced glutathione (GSH) levels, suppresses the expression of the E6 oncoprotein, activates the expression of the p53 tumor suppressor, and reduces the mitochondrial potential of tumor cells. Binase-induced disruption of the integrity of the mitochondrial membrane is a signal for activation of the mitochondrial apoptosis pathway. [ABSTRACT FROM AUTHOR]

It is thought to be risk-free, environmentally benign, and safe for biological processes to produce zinc oxide nanoparticles from renewable resources. This study examined Cassia javanica's ability to create ZnONPs. The generated ZnONPs were analyzed using a variety of techniques, such as TEM, FTIR spectroscopy, UV-Vis spectroscopy, and XRD analysis. The antibacterial potential of ZnONPs has been investigated using both Agar well diffusion and microtitreplate (MTP) methods. One method used to evaluate ZnONPs' capacity to scavenge free radicals at different concentrations was the DPPH method. The permanent zinc oxide (ZnO) shape and the naturally occurring crystal structure of ZnONPs were validated by the XRD data. ZnONPs showed antibacterial activity with MICs of 31.7 μg/mL toward Bacillus subtilis, 62.5 μg/mL for Salmonella typhimurium, Escherichia coli while Clostridium sporogenes and Bacillus pumilus was 125μg/mL. Furthermore, ZnONPs demonstrated a range of antibiofilm activities toward Staphylococcus aureus (MRSA). ZnONPs showed an intriguing antioxidant capacity, achieving IC50 of 109.3 μg/ml μg/mL. Additionally, ZnONPs demonstrated low toxic effect on Vero cell with IC50 154.01 μg/mL as well as possible anticancer action when applied to the carcinoma cell lines HepG2 with IC50 of 47.48 μg/mL. Furthermore, ZnONPs at 62.5 μg/mL had a promising antiviral impact against HSV1 and COX B4, with antiviral activities of 75.4% and 65.8%, respectively. [ABSTRACT FROM AUTHOR]

Plastic pollution has quadrupled over the past years and has become a global concern due to its direct impact on life forms. The present study analysed whether the plastic debris in aquatic environments could act as the substratum for the antimicrobial-resistant (AMR) bacteria to form biofilm for survival. We have collected various plastic debris (n = 32) from six sites of the Periyar River, the drinking water source for the entire city and one of the most polluted rivers in Kerala (India). The chemical composition of plastics was screened via FTIR analysis and found that they comprised two types, viz., polyethylene and polypropylene. Bacteria isolated from the samples were screened for the AMR characteristics towards eight different classes of antibiotics. All isolates showed 100% resistance towards colistin and obtained the MAR index value of 0.1–0.4 range. Six representative bacterial isolates with high multiple antibiotic resistance (MAR) index were selected and identified by 16sRNA sequencing as Lysinibacillus mangiferihumi, Bacillus pumilus, Bacillus safensis, Bacillus cereus, Bacillus altitudins and Bacillus pumilus. In vitro biofilm formation was experimented on the purchased plastic samples in artificial media and river water using two selected strains, Bacillus pumilus and Bacillus cereus. Significant variations were observed in biofilm growth in different media (P < 0.05) regardless of plastic types (P > 0.05). The extracellular polymeric substances (EPS) and the characteristic holes on the surface morphology were visualized in SEM analysis, thus indicating the conditioning of the plastics by the isolates for biofilm formation. [ABSTRACT FROM AUTHOR]

Cadmium (Cd) is a well-known pollutant in agricultural soil, affecting human health through the food chain. To combat this issue, Ca + Mg (25 mg L-1) nanocomposite and Bacillus pumilus, either alone or combined, were applied to rice plants under Cd (5 mg kg-1, 10 mg kg-1) contamination. In our study, growth and yield traits demonstrated the beneficial influence of Ca + Mg and B. pumilus application in improving rice defense mechanism by reducing Cd stress. Combined Ca + Mg and B. pumilus application increased SPAD (15), total chlorophyll (18), chlorophyll a (11), chlorophyll b (22), and carotenoids (21%) with Cd (10 mg kg-1), compared to the application alone. Combined Ca + Mg and B. pumilus application significantly regulated MDA (15), H2O2 (13), EL (10), and O2 •- (24%) in shoots under Cd (10 mg kg-1), compared to the application alone. Cd (10 mg kg-1) increased the POD (22), SOD (21), APX (12), and CAT (13%) in shoots with combined Ca + Mg and B. pumilus application, compared to the application alone. Combined Ca + Mg and B. pumilus application significantly reduced Cd accumulation in roots (22), shoots (13), and grains (20%) under Cd (10 mg kg-1), compared to the application alone. Consequently, the combined application of Ca + Mg and B. pumilus is a sustainable solution to enhance crop production under Cd stress. [ABSTRACT FROM AUTHOR]

Selenium nanoparticles (SeNPs) are used in several sectors as antitumor, antimicrobial, and environmental adsorbents. Thus, the present research objective was the production of bacterial-SeNPs as an active and environmentally-friendly antibacterial and adsorbent agents and application into novel nanocomposite filter. From a total of 25 samples (soil, wastewater, and water) obtained from different locations in Egypt, 60 selenium-resistant bacterial isolates were obtained (on a mineral salt medium supplemented with selenium ions). After screening (based on the conversion of selenium from ionic form to nanoform), a superior bacterial isolate for SeNPs formation was obtained and molecular identified as Bacillus pumilus isolate OR431753. The high yield of SeNPs was noted after optimization (glucose as carbon source, pH 9 at 30 °C). The produced SeNPs were characterized as approximately 15 nm-diameter spherical nanoparticles, in addition to the presence of organic substances around these particles like polysaccharides and aromatic amines (protein residues). Also, they have antibacterial activity increased after formation of nanocomposite with nano-chitosan (SeNPs/NCh) against several pathogens. The antibacterial activity (expressed as a diameter of the inhibitory zone) averaged between 2.1 and 4.3, 2.7 and 4.8 cm for SeNPs and SeNPs/NCh, respectively compared with 1.1 to 1.8 cm for Amoxicillin. The produced nanoselenium/chitosan was used as a biofilter to remove mercury (Hg) and AgNPs as model chemicals with serious toxicity and potential pollutant for water bodies in many industries. The new SeNPs/NCh biofilter has proven highly effective in individually removing mercury and AgNPs from their synthetic wastewaters, with an efficiency of up to 99%. Moreover, the removal efficiency of AgNPs stabilized at 99% after treating them with the syringe filter-Se nanocomposite for 4 cycles of treatment (5 min each). [ABSTRACT FROM AUTHOR]

The purpose of this study was to isolate, identify, and evaluate the antibacterial and probiotic potential of bacteria from honeydew honey collected in Poland. Isolates (189 colonies from 10 honey samples) were evaluated for their antimicrobial activity against Staphylococcus aureus, Bacillus cereus, Escherichia coli, and Yersinia enterocolitica, and then identified by MALDI-TOF-MS. The isolates with the greatest antimicrobial properties were screened for their probiotic potential. The total number of bacteria isolated from honey did not exceed the value of 2.5 × 102 CFU/mL. The Bacillus pumilus/altitudinis, B. licheniformis, and Bacillus cereus groups were the dominant identified bacteria. Almost 16% of the isolates expressed antibacterial potential against three pathogenic bacteria, over 20% against two, while almost 34% of the isolates did not inhibit any. The survival rate of the isolates under gastrointestinal tract conditions was higher after 4 h of exposure to bile salts (>60% survival rate for 66.66% of the isolates), while at pH 2.0, it was lower (>50% survival rate for 44% of the isolates). The most resistant isolate B. pumilus/altitudinis survived at a rate of 77% at low pH and 108% with bile salts. These results confirmed that honeydew honey is a promising reservoir of bacteria that produces metabolites with antimicrobial and probiotic potential. [ABSTRACT FROM AUTHOR]

In this study, the effect of inoculation with PGPR on growth, sulfur and DL-sulforaphane content of broccoli seedlings was evaluated. Rhizobacterial strains of Bacillus pumilus R44, Pseudomonas tolaasii P61 and P. tolaasii A46 were screened for total indole production and P solubilization. Broccoli seedlings were inoculated with these strains in order to establish a greenhouse experiment. Seedlings were harvested 30 days after inoculation to determine fresh weight, dry weight, root volume, height, and relative chlorophyll content, also S uptake and sulforaphane content. Treatment with PGPR resulted in an increase in almost all the variables evaluated compared to the control. Outstanding results were obtained with the A46 and R44 strains, being 18 and 49% higher for sulforaphane content. P61 was the strain with the best results related to indole production (384% higher than A46), phosphate solubilization (287% higher than A46), and root volume (44% higher than control). There were no significant differences in relative chlorophyll content and sulfur accumulation among the treatments. This is the first work that showed the positive effect of PGPR on sulforaphane content in broccoli seedlings, a compound reported with anticancer activity. [ABSTRACT FROM AUTHOR]

This study investigated the effect of pH on poly-γ-l-diaminobutanoic acid (γ-PAB) production by Bacillus pumilus in batch fermentation. In the natural fermentation where pH was not controlled, pH decreased from initial 7.0 to 3.0 in 18 h and γ-PAB production was 428.6 mg/L. In the pH-controlled fermentation, B. pumilus tended to proliferation at higher pH, while γ-PAB synthesis was favorable at lower pH, in which the optimal pH for γ-PAB production was 4.2, and γ-PAB yield reached 2284.5 mg/L. Adopting a pH shock strategy which lasted 9 h in the pre-fermentation phase, biomass (OD600) and γ-PAB yield of B. pumilus were obtained as 61.3 and 2794.6 mg/L, respectively, which were 10.8% and 22.4% higher than those in batch fermentation without pH shock. Subsequent fermentation of repeated pH shocks showed that a further higher productivity could be achieved, in which the final OD600 reached 65.1, and γ-PAB production reached as high as 3482.3 mg/L, which were increased by 6.2% and 17.1% compared with those in single pH shock, respectively. This study demonstrated that B. pumilus can synthesize more γ-PAB at suboptimal pH and provided a novel approach to regulate γ-PAB synthesis. [ABSTRACT FROM AUTHOR]

Mycotoxins, toxic compounds produced by fungi, pose significant risks to food safety and human health. This study investigates the bio-detoxification potential of 238 strains of lactic acid bacteria (LAB) and Bacillus spp., previously isolated from cereals (including mycotoxin-contaminated grains), against the emerging mycotoxin, enniatin B (ENB), and the prevalent mycotoxin, deoxynivalenol (DON). Out of the tested strains, 26 demonstrated notable mycotoxin reduction capabilities, including 2 Bacillus pumilus and 24 Bacillus licheniformis strains. B. licheniformis strains MA572, MA695, MA696, TR174a, TR284, TR363, and TR466a degraded ENB to levels below the detection limit, and six strains reduced DON by 30–35%; B. licheniformis TR251b and TR374 showed the highest DON reduction with 35.7%. The most promising strains for bio-detoxification were B. licheniformis TR284, which achieved a 100% reduction in ENB and a 28.6% reduction in DON and B. licheniformis TR388 with a 97.5% reduction in ENB and a 31.9% reduction in DON. None of the tested LAB strains significantly reduced either mycotoxin. These findings highlight the promising potential of B. licheniformis strains in bio-detoxifying mycotoxin-contaminated cereal products. Further research into the underlying detoxification mechanisms and safety aspects is essential to develop effective bio-detoxification strategies for enhancing food safety. [ABSTRACT FROM AUTHOR]

Abstract Stem nematode (Ditylenchus destructor Thorne) is considered one of the most economically devastating species affecting sweet potato production. Biocontrol offers a sustainable strategy for nematode control. This study conducted a pot experiment to evaluate the biocontrol efficacy of Paecilomyces lilacinus CS-Z and Bacillus pumilus Y-26 against the stem nematode, as well as to examine their influence on the bacterial communities in the sweet potato rhizosphere. The findings indicated that B.pumilus Y-26 and P.lilacinus CS-Z exhibited respective suppression rates of 82.9% and 85.1% against the stem nematode, while also stimulating sweet potato plant growth. Both high-throughput sequencing and Biolog analysis revealed distinct impacts of the treatments on the bacterial communities. At the phylum level, B.pumilus Y-26 enhanced the abundance of Actinobacteria but reduced the abundance of Cyanobacteria, with P.lilacinus CS-Z exhibiting similar effects. Additionally, the treatment with B.pumilus Y-26 resulted in increased abundances of Crossiella, Gaiella, Bacillus, and Streptomyces at the genus level, while the treatment with P.lilacinus CS-Z showed increased abundances of Crossiella and Streptomyces. In contrast, the abundance of Pseudarthrobacter was reduced in the treatment with B.pumilus Y-26. Conversely, the application of the nematicide fosthiazate exhibited minor influence on the bacterial community. The findings indicated that the application of P.lilacinus CS-Z and B.pumilus Y-26 led to an increase in the relative abundances of beneficial microorganisms, including Gaiella, Bacillus, and Streptomyces, in the rhizosphere soil. In conclusion, P.lilacinus CS-Z and B.pumilus Y-26 demonstrated their potential as environmentally friendly biocontrol agents for managing stem nematode disease of sweet potato.

Abstract Selenium nanoparticles (SeNPs) are used in several sectors as antitumor, antimicrobial, and environmental adsorbents. Thus, the present research objective was the production of bacterial-SeNPs as an active and environmentally-friendly antibacterial and adsorbent agents and application into novel nanocomposite filter. From a total of 25 samples (soil, wastewater, and water) obtained from different locations in Egypt, 60 selenium-resistant bacterial isolates were obtained (on a mineral salt medium supplemented with selenium ions). After screening (based on the conversion of selenium from ionic form to nanoform), a superior bacterial isolate for SeNPs formation was obtained and molecular identified as Bacillus pumilus isolate OR431753. The high yield of SeNPs was noted after optimization (glucose as carbon source, pH 9 at 30 °C). The produced SeNPs were characterized as approximately 15 nm-diameter spherical nanoparticles, in addition to the presence of organic substances around these particles like polysaccharides and aromatic amines (protein residues). Also, they have antibacterial activity increased after formation of nanocomposite with nano-chitosan (SeNPs/NCh) against several pathogens. The antibacterial activity (expressed as a diameter of the inhibitory zone) averaged between 2.1 and 4.3, 2.7 and 4.8 cm for SeNPs and SeNPs/NCh, respectively compared with 1.1 to 1.8 cm for Amoxicillin. The produced nanoselenium/chitosan was used as a biofilter to remove mercury (Hg) and AgNPs as model chemicals with serious toxicity and potential pollutant for water bodies in many industries. The new SeNPs/NCh biofilter has proven highly effective in individually removing mercury and AgNPs from their synthetic wastewaters, with an efficiency of up to 99%. Moreover, the removal efficiency of AgNPs stabilized at 99% after treating them with the syringe filter-Se nanocomposite for 4 cycles of treatment (5 min each).

Culicoides oxystoma Kieffer (Diptera: Ceratopogonidae) has been vectoring several arboviruses, protozoa and nematodes, leading to mortality and morbidity of livestock and wild ruminants in the tropics and subtropics. Insight into the bacterial communities associated with the vector species must be worked out. This work tries to inventorize the bacterial communities associated with this important vector species. Acquisition of gut microbiota may be the parental origin, while some are obtained through feeding during larval stages. Culicoides oxystoma possesses semi‐aquatic life cycle strategies for egg‐laying and larval survival. The bacteria associated with C. oxystoma were compared throughout (i) life stages: egg, larval instars, pupa, adult: male and female obtained from laboratory colony; (ii) field‐collected adult: male and age‐graded females; and (iii) natural breeding substrate and artificial rearing substrate. The culture‐dependent bacteria were identified by Sanger sequencing of 16S rRNA, and haemolytic bacteria were screened on blood agar. Results show that Firmicutes and Proteobacteria are the predominant Phyla, of which Bacillus spp. was the most abundant across the life stages. Across the life history, Bacillus cereus, Bacillus pumilus, Bacillus tropicus, Lysinibacillus sp. and Paenibacillus sp. were retrieved routinely. Bacillus cereus and Alcaligenes faecalis were detected in the lab‐reared specimens and shared between the natural breeding site and rearing medium. From the adults trapped across two locations, B. cereus, Bacillus flexus, A. faecalis, Enterococcus faecium and Pseudomonas sp. were isolated. The bacterial species associated with this vector may influence various physiological traits, such as vectorial capacity, digestion and larval development, which need further investigation. [ABSTRACT FROM AUTHOR]

The list of standard abbreviations for JDS is available at adsa.org/jds-abbreviations-24. Nonstandard abbreviations are available in the Notes. Objectives were to evaluate the effects of a multistrain Bacillus -based (Bacillus subtilis and Bacillus pumilus blend) direct-fed microbial (DFM) on production, metabolism, inflammation biomarkers and gastrointestinal tract (GIT) permeability during and following feed restriction (FR) in mid-lactation Holstein cows. Multiparous cows (n = 36; 138 ± 53 DIM) were randomly assigned to 1 of 3 dietary treatments: (1) control (CON; 7.5 g/d rice hulls; n = 12), (2) DFM10 (10 g/d Bacillus DFM, 4.9 × 109 cfu/d; n = 12) or 3) DFM15 (15 g/d Bacillus DFM, 7.4 × 109 cfu/d; n = 12). Before study initiation, cows were fed their respective treatments for 32 d. Cows continued to receive treatments during the trial, which consisted of 3 experimental periods (P): P1 (5 d) served as baseline for P2 (5 d), during which all cows were restricted to 40% of P1 DMI, and P3 (5 d), a "recovery" where cows were fed ad libitum. On d 4 of P1 and on d 2 and 5 of P2, GIT permeability was evaluated in vivo using the oral paracellular marker Cr-EDTA. As anticipated, FR decreased milk production, insulin, glucagon, and BUN but increased nonesterified fatty acids. During recovery, DMI rapidly increased on d 1 then subsequently decreased (4.9 kg) on d 2 before returning to baseline, whereas milk yield slowly increased but remained decreased (13%) relative to P1. The DFM10 cows had increased DMI and milk yield relative to DFM15 during P3 (10%). Overall, milk lactose content was increased in DFM cows relative to CON (0.10 percentage units), and DFM10 cows tended to have increased lactose yield relative to CON and DFM15 during P3 (8% and 10%, respectively). No overall treatment differences were observed for other milk composition variables. Circulating glucose was quadratically increased in DFM10 cows compared with CON and DFM15 during FR and recovery. Plasma Cr area under the curve was increased in all cows on d 2 (9%) and 5 (6%) relative to P1. Circulating LPS binding protein (LBP), serum amyloid A (SAA), and haptoglobin (Hp) increased in all cows during P2 compared with baseline (31%, 100%, and 9.0-fold, respectively). Circulating Hp concentrations continued to increase during P3 (274%). Overall, circulating LBP and Hp tended to be increased in DFM15 cows relative to DFM10 (29% and 81%, respectively), but no treatment differences were observed for SAA. Following feed reintroduction during P3, fecal pH initially decreased (0.62 units), but returned to baseline levels whereas fecal starch markedly increased (2.5-fold) and remained increased (82%). Absolute quantities of a fecal Butyryl-CoA CoA transferase (but) gene associated with butyrate synthesis, collected by fecal swab were increased in DFM10 cows compared with CON and DFM15 cows. In summary, FR increased GIT permeability, caused inflammation, and decreased production. Feeding DFM10 increased some key production and metabolism variables and upregulated a molecular biomarker of microbial hindgut butyrate synthesis, while DFM15 appeared to augment immune activation. [ABSTRACT FROM AUTHOR]

Simple Summary: Assessing the quality of raw milk is crucial for humans' well-being. This study aimed to evaluate the bacterial composition and antimicrobial resistance in the tea cup liners of dairy cow milking equipment. The objective was to determine the potential risk of bacterial contamination in raw milk during the milking process and the transmission of antimicrobial-resistant bacteria through milk sources. The study found that Bacillus spp. were the most prevalent bacteria identified on the teat cup liners. A significant proportion of these bacteria showed significant resistance to various antibiotics, such as lincomycin, sulfadiazine, and streptomycin. Multiple antimicrobial resistance genes were also detected, indicating the potential transmission of antimicrobial-resistant bacteria in the milk samples. These findings underscore the imperative to enhance disinfection protocols within dairy farms and milking equipment. The results accentuate the enduring peril of disseminating antimicrobial-resistant bacteria via contaminated milk sources. Taking heed of this finding will contribute to the sustainable development of the dairy industry while mitigating the hazards associated with food safety. Increasing awareness of the potential dissemination of antimicrobial-resistant bacteria through dairy products can foster the implementation of appropriate measures to ensure the secure production and consumption of such products, thereby safeguarding public health and welfare. Global milk consumption exceeds 800 million tons a year and is still growing. Milk quality and its products are critical to human health. A teat cup makes direct contact with the cow's teats during milking and its cleanliness is very important for the quality of raw milk. In this study, the microorganism from post-milking teat cup liners were collected from six dairy farms in Shandong Province of China, the bacterial species were identified using microbial mass spectrometry, the minimum inhibitory concentrations of the isolated strains against ten antimicrobial agents were determined using the broth microdilution method, and the antimicrobial resistance genes were detected by PCR. The results indicated that the most frequently isolated bacteria in this study were Bacillus licheniformis (39/276, 14.13%), followed by Bacillus pumilus (20/276, 7.25%), Bacillus cereus (17/276, 6.16%), and Bacillus subtili (16/276, 5.80%). The isolates exhibited the highest average resistance to lincomycin (87.37%), followed by sulfadiazine (61.05%) and streptomycin (42.63%); the highest detection rate of resistance genes was Sul1 (55.43%), followed by ant(4') (51.09%), tet(M) (25.36%), blaKPC (3.62%) and qnrS (3.62%). These findings imply the necessity for enhanced measures in disinfecting cow udders and milking equipment, highlighting the persistently challenging issue of antimicrobial resistance in Shandong Province. [ABSTRACT FROM AUTHOR]

We studied the occurrence of endophytic fungi and bacteria in the roots of F. gigantea, a woodland perennial grass common throughout Europe and temperate Asia. The taxonomic assignment was carried out according to the isolate's colony and cytological species morphotype characteristics and confirmed by the assessment of the standard DNA sequences, ITS, RPB2, SSU, and TEF1-a for fungi and 16S rDNA for bacteria. Our study has shown that F. gigantea roots are the habitat to a wide range of fungi and bacteria. The occurrence of fungal structures was determined in ~40% of the roots examined by Trypan Blue staining. In a surface-sterile root-cutting culture on PDA medium, we obtained isolates of six endophytic fungi species: four members of Ascomycota—Alternaria alternata, Cadophora fastigiata, Chaetomium funicola, and Microdochium bolleyi—and two of Basidiomycota—Coprinellus sp. and Sistotrema brinkmannii. In addition, we report bacteria co-occurring endophytically in the roots of this grass. The Firmicutes group was the most prevalent, consisting of four Gram-positive, endospore-forming bacteria taxa. The isolates were identified as Bacillus pumilus, Bacillus sp., Lysinibacillus sp., and Priestia aryabhattai. Moreover, two Gram-negative bacteria were detected—Kosakonia sp. (Proteobacteria) and Pedobacter sp. (Bacteroidetes). Thus, applying the isolate-culture approach, we identified a set of microorganisms in the roots of a typical grass native to the deciduous forest floor. The functional roles of these endophytes are diverse, and many of them, saprotrophs and decomposers of wood and plant debris, are linked to the decomposition of organic matter. This is the first detailed report on fungal and bacterial endophytes inhabiting the roots of F. gigantea. This study fills in a research gap on endophytes associated with the below-ground parts of Festuca spp., hitherto extensively studied for Epichloë/Neotyphodium associations in their foliar parts. [ABSTRACT FROM AUTHOR]

Acute hepatopancreatic necrosis disease, caused by Vibrio parahaemolyticus strains carrying the pirA and pirB toxin genes (VpAHPND), has been causing great economic losses in Asia and America in the shrimp farming industry. Numerous strains are resistant to antibiotics. However, supplementation with probiotic antagonists has become a more desirable treatment alternative. Fourteen strains of microorganisms were assessed for their potential to inhibit VpAHPND in vitro activity. The bacteria with the highest activity were challenged with VpAHPND-infected Pacific white shrimp Litopenaeus vannamei. Furthermore, the genomic characteristics of probiotic bacteria were explored by whole-genome sequencing. We identified the Sonora strain as Bacillus pumilus, which possesses positive proteolytic and cellulolytic activities that may improve shrimp nutrient uptake and digestion. Challenge trials showed a low cumulative mortality (11.1%). B. pumilus Son has a genome of 3,512,470 bp and 3734 coding sequences contained in 327 subsystems. Some of these genes are related to the biosynthesis of antimicrobial peptides (surfactins, fengycin, schizokinen, bacilibactin, and bacilysin), nitrogen and phosphorus metabolism, and stress response. Our in vitro and in vivo findings suggest that B. pumilus Sonora has potential as a functional probiotic. [ABSTRACT FROM AUTHOR]

We study the carbonic anhydrase (CA) pathway using autochthonous CA-producing bacteria as a means of inducing calcite precipitation, which acts as a biocement to improve the engineering soil properties. Forty different microbial strains producing CA were isolated from the foundation soil of a railway embankment in Prickwillow, UK. Three of the best CA-producing strains were selected and identified by DNA sequencing as Bacillus licheniformis, Bacillus toyonensis and Bacillus pumilus with CA activity values respectively of 1.79 U/ml, 1.42 U/ml and 1.55 U/ml. To optimise the treatments, we investigated the effect of pH, temperature, zinc co-factor and cementation solution molarity on the growth and CA activity and bioprecipitates, with CO2 added in the form of bicarbonate. Scanning electron microscope (SEM) analysis of the bioprecipitates showed that these had characteristic morphologies of calcite and vaterite crystals. The formation of calcite was further corroborated by FT-IR and Raman analysis of bioprecipitates. The precultured bacteria were injected into the fine-grained soil together with cementation solution. Unconfined compressive strength in treated soil increased up to 1 MPa, and its calcium carbonate content increased by 2.78%. This, as well as the stability of the treated soil upon water immersion, proved the biocementation of the fine-grained soil. These findings suggest the potential of employing the CA biocementation route for soil stabilisation pending further development of the technique. [ABSTRACT FROM AUTHOR]

The current study attempted to develop a bacterial consortium which can degrade chlorpyrifos pesticide efficiently. Five morphologically different bacterial strains, capable to tolerate high chlorpyrifos concentration, were isolated from pesticide-contaminated soil. Further, possible combinations of the isolates were formed and each combination was screened and chlorpyrifos degradation percentage and half-lives were calculated. The consortium CON11, comprising of four isolates (CP28, CP30, CP31, and CP34) showed the highest chlorpyrifos degradation. The individual independent variables of chlorpyrifos degradation by CON11 viz. temperature (°C), pH, incubation time (days) and pesticide concentration (ppm) were optimized using one-factor at-a-time experiments. The consortium isolates were then identified by the molecular tools 16S rRNA gene typing and the sequences were submitted to GenBank. The four bacterial isolates thus identified are Bacillus amyloliquefaciens CP28 (MH667455), Bacillus pumilus CP30 (MH667456), Bacillus marisflavi CP31 (MN715880), and Bacillus subtilis CP34 (MH667458). These isolates which comprise CON11 consortium could degrade 91% of 100 ppm chlorpyrifos after 6 days of incubation at 30 °C and pH 7 with a half-life of 1.73 days indicating the promising potential of the consortium, CON11, in the cleanup of chlorpyrifos contaminated sites. [ABSTRACT FROM AUTHOR]

This study utilized Aspergillus flavus to produce selenium nanoparticles (Se-NPs) in an environmentally friendly and ecologically sustainable manner, targeting several medicinal applications. These biosynthesized Se-NPs were meticulously characterized using X-ray diffraction (XRD), Fourier-transform infrared (FT-IR) spectroscopy, transmission electron microscope (TEM), and UV–visible spectroscopy (UV), revealing their spherical shape and size ranging between 28 and 78 nm. We conducted further testing of Se-NPs to evaluate their potential for biological applications, including antiviral, anticancer, antibacterial, antioxidant, and antibiofilm activities. The results indicate that biosynthesized Se-NPs could be effective against various pathogens, including Salmonella typhimurium (ATCC 14028), Bacillus pumilus (ATCC 14884), Staphylococcus aureus (ATCC 6538), Clostridium sporogenes (ATCC 19404), Escherichia coli (ATCC 8739), and Bacillus subtilis (ATCC 6633). Additionally, the biosynthesized Se-NPs exhibited anticancer activity against three cell lines: pancreatic carcinoma (PANC1), cervical cancer (Hela), and colorectal adenocarcinoma (Caco-2), with IC50 values of 177, 208, and 216 μg/mL, respectively. The nanoparticles demonstrated antiviral activity against HSV-1 and HAV, achieving inhibition rates of 66.4% and 15.1%, respectively, at the maximum non-toxic concentration, while also displaying antibiofilm and antioxidant properties. In conclusion, the biosynthesized Se-NPs by A. flavus present a promising avenue for various biomedical applications with safe usage. [ABSTRACT FROM AUTHOR]

Due to the emergence of drug-resistant microorganisms, the search for broad-spectrum antimicrobial compounds has become extremely crucial. Natural sources like plants and soils have been explored for diverse metabolites with antimicrobial properties. This study aimed to identify microorganisms from agricultural soils exhibiting antimicrobial effects against known human pathogens, and to highlight the chemical space of the responsible compounds through the computational metabolomics-based bioprospecting approach. Herein, bacteria were extracted from soil samples and their antimicrobial potential was measured via the agar well diffusion method. Methanolic extracts from the active bacteria were analyzed using the liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF-MS) technique, and the subsequent data was further analyzed through molecular networking approach which aided in identification of potential anti-microbial compounds. Furthermore, 16S rRNA gene sequencing enabled identification of the active bacterial isolates, where isolate 1 and 2 were identified as strains of Bacillus pumilus, whilst isolate 3 was found to be Bacillus subtilis. Interestingly, isolate 3 (Bacillus subtilis) displayed wide-ranging antimicrobial activity against the tested human pathogens. Molecular networking revealed the presence of Diketopiperazine compounds such as cyclo (D-Pro-D-Leu), cyclo (L-Tyr-L-Pro), cyclo (L-Pro-D-Phe), and cyclo (L-Pro-L-Val), alongside Surfactin C, Surfactin B, Pumilacidin E, and Isarrin D in the Bacillus strains as the main anti-microbial compounds. The application of the molecular networking approach represents an innovation in the field of bio-guided bioprospection of microorganisms and has proved to be an effective and feasible towards unearthing potent antimicrobial compounds. Additionally, the (computational metabolomics-based) approach accelerates the discovery of bioactive compounds and isolation of strains which offer a promising avenue for discovering new clinical antimicrobials. Finally, soil microbial flora could serve an alternative source of anti-microbial compounds which can assist in the fight against emergence of multi-drug resistance bacterial pathogens. [ABSTRACT FROM AUTHOR]

Ciprofloxacin (CPX), is a broad-spectrum fluoroquinolone antibiotic widely used to treat bacterial infections in humans and animals. In recent years, ecological problems caused by the disposal of CPX-waste product have attracted increasing attention. This study aimed to evaluate the ability of different bacteria to remove CPX from water. The ability of five bacterial strains namely; Acenitobacter sp., Acenitobacter lwofii, Bacillus. pumilus, Burkholderia phytofirmans, and Mesorihizobium sp. were examined in this study. Tests were performed on Luria Bertani (LB) as well as on Mueller-Hinton (MH) agar at different CPX concentrations (20-100 mg L-1). All five tested bacterial strains showed growth on LB as well as on MH agar plates at all concentrations between 20 to 100 mg L-1 CPX. Three bacterial species: Acenitobacter lwofii, Bacillus pumilus, and Mesorihizobium sp. were reported more resistant to CPX of the five strains tested. In a shake flask study, the strain B. pumilus initially showed higher degradation rate (75.53%) followed by Acenitobacter lwofii (72.3%), Mesorihizobium sp. (70.32), Burkholderia phytofirmans (67.10%), and Acenitobacter sp. (63.2%) at low CPX concentration (5 mg L-1) in minimal saline media. However, Mesorihizobium sp. showed higher degradation efficiency (76.32%) at a higher CPX concentration (10 mg L-1). A group of three bacterial strains, A. lowfi, B. pumilus, and Mesorihizobium sp. showed good CPX degradation rate (95.4%), and achieved higher degradation efficiency than the single strains at 20 mg L-1 CPX concentration. Overall, the results suggest that using a combination of bacteria may be a useful tool for the remediation of antibiotic pollution. In addition, this study reveals the ability of previously isolated bacteria to biodegrade CPX as a sole source of carbon, providing new insights into the biodegradation of CPX. [ABSTRACT FROM AUTHOR]

The use of nitrogen-fixing bacteria in leguminous plants is a widespread approach, and the exploration of symbiotic bacteria that are efficient in biological nitrogen fixation (BNF) continues to be explored. The aim of the present study was to evaluate the effect of inoculation and coinoculation of mung bean plants with different combinations of Bradyrhizobium (B. elkanii BR 2003, B. pachyrhizi BR 3262, B. yuanmingense BR 3267, B. paxllaeri BR 10398 and B. icense BR 10399), Azospirillum baldaniorum (Sp245) and Bacillus pumilus (UFPEDA 472) on the contribution of biomass, the concentration of nitrogen (N) compounds and BNF. The experiment was carried out under greenhouse conditions with pots containing washed and autoclaved sand. Mung bean seeds were inoculated or coinoculated (double or triple) with Bradyrhizobium, A. baldaniorum and/or Bacillus pumilus, and one absolute control (not inoculated) was used. The experimental design was a completely randomized design with 21 treatments harvested in two different periods (flowering and pod maturation). Inoculation and coinoculation positively influenced the number of nodules, shoot dry weight, N accumulated, total N content and inorganic and organic compounds (free ammonia, nitrate, ureides and leghemoglobin), indicating that there was efficiency in BNF and synergistic interaction between the bacteria used and the mung bean plants. Inoculation with Bradyrhizobium species and the combination of these strains with A. baldaniorum and Bacillus pumilus positively influenced N fixation and metabolism in mung bean plants, especially when B. elkanii BR 2003 and B. pachyrhizi BR 3262 were used. [ABSTRACT FROM AUTHOR]

Poly‐L‐lactic acid (PLLA) is currently the most abundant bioplastic; however, limited environmental biodegradability and few recycling options diminish its value as a biodegradable commodity. Enzymatic recycling is one strategy for ensuring circularity of PLLA, but this approach requires a thorough understanding of enzymatic mechanisms and protein engineering strategies to enhance activity. In this study, we engineer PLLA depolymerizing subtilisin enzymes originating from Bacillus species to elucidate the molecular mechanisms dictating their PLLA depolymerization activity and to improve their function. The surface‐associated amino acids of two closely related subtilisin homologues originating from Bacillus subtilis (BsAprE) and Bacillus pumilus (BpAprE) were compared, as they were previously engineered to have nearly identical active sites, but still varied greatly in PLLA depolymerizing activity. Further analysis identified several surface‐associated amino acids in BpAprE that lead to enhanced PLLA depolymerization activity when engineered into BsAprE. In silico protein modelling demonstrated increased enzyme surface hydrophobicity in engineered BsAprE variants and revealed a structural motif favoured for PLLA depolymerization. Experimental evidence suggests that increases in activity are associated with enhanced polymer binding as opposed to substrate specificity. These data highlight enzyme adsorption as a key factor in PLLA depolymerization by subtilisins. [ABSTRACT FROM AUTHOR]

As we know the recent pandemic, coronavirus disease (COVID-19) due to SARS CoV-2 virus has led to an increase in the consumption of various drugs as medicines by the patients. Paracetamol (acetaminophen, APAP) act as an emerging contaminant classified among the class pharmaceutical and personal care pollutant (PPCP) and is detected in wastewater and sewage systems. The enrichment culture approach was used for the isolation APAP-degrading bacterium wastewater sample. Microscopic examination, biochemical and 16S rRNA sequence analysis showed that the isolate PYP-2 belongs to the Bacillus pumilus strain. Shake flask and batch culture degradation studies have shown that the strain can degrade APAP. Further, the response surface methodology (RSM) plot was used to know the best physical condition for biodegradation by optimization study. The optimum pH of 5.0, temperature of 30 °C, agitation speed of 146 rpm, and APAP 267 mg/L concentration were reported for PYP-2-based degradation. Bacterial biomass kinetic analysis was performed at the best physical condition, and the results showed that the specific growth rate (µ) was 713 mg/L. Oxalic acid, 2-isopropyl-5-methyl cyclohexanone, and phenothiazine were the intermediates of the APAP degradation pathway detected by the GC-MS chromatogram peaks. Therefore, this research has shown that Bacillus pumilus strain PYP-2 has the metabolic capacity to biodegrade APAP, providing new tools for bioremediation. [ABSTRACT FROM AUTHOR]

Petroleum serves as a vital energy source, but extensive exploration can lead to frequent accidents, resulting in soil contamination from oil spills, which poses risks to both ecosystems and human health. Bioremediation emerges as a potent method for addressing polluted environments, involving the deliberate selection and application of microorganisms. In the present study, bacteria present in the petroleum oil contaminated soil were isolated by enrichment culture technique using Bushnall Hass (BS) media supplemented with petroleum oil as a sole carbon source. The isolated bacterial strains were characterized by morphological and biochemical tests and further characterized through molecular tools ie, sequencing method. Three bacterial isolates were morphologically characterized as gram-positive, spherical, cocci, and rod shaped further 16S rRNA sequence analysis revealed that the isolates are closely related to Staphylococcus hominis, Staphylococcus sp., and Bacillus pumilus, respectively. The optimum growth was at pH 7.0 and temperature 37 °C. All strains were susceptible to various antibiotics, and resistant to various heavy metals and have no antagonistic effect with oil degrading bacteria. All strains were grown in five different concentrations of petroleum oil (1%, 1.50%, 2%, 2.50%, and 3%), respectively. Study revealed that all the isolates showed their degradation efficacy in 3% v/v petroleum oil. So, this research clearly demonstrates that the isolated bacterial strains could be effectively used for bioremediation of petroleum-contaminated areas. [ABSTRACT FROM AUTHOR]

To the best of our knowledge, this paper presents the first report on the microbial status of wild bees (A. dorsata and A. florea) honey collected from different northern states of India. In total, 17 bacterial morphotypes (9 from A. dorsata and 8 from A. florea) were recovered. The bacterial load (log CFU/g) was observed to be maximum (4.01) in A. dorsata honey from Sirmour-Puruwala whereas, lowest load (3.49) was recorded in Punjab-Rupnagar honey. Likewise, A. florea honey from Sirmour-Kala Amb and Punjab-Doraha were recorded with maximum and minimum bacterial loads of 4.17 and 3.40, respectively. Observance of lower bacterial load than hazardous limit ensured its safety for human consumption. Three most dominant bacteria viz., AD1, HF7 and AF4 were identified as Micrococcus endophyticus (MT938911), Bacillus subtilis subsp. stercoris (MT764923) and Bacillus pumilus (MT764923), respectively through 16S rRNA ribotyping. Moreover, the inhibitory activity of honey was good to excellent against Salmonella typhi NCTC 786 and fair to good against Bacillus subtilis ATCC 6633, Pseudomonas aeruginosa ATCC 10662 and Klebsiella pneumoniae ATCC 13883. The Escherichia coli ATCC 1041 was the most sensitive bacterium among all. These results justify wild bees honey as a natural remedy for curing and preventing numerous human ailments. [ABSTRACT FROM AUTHOR]

ABSTRACT: The widespread use of antibiotics causes the development of antibiotic-resistant bacterial strains, which have a severe impact on poultry productivity and human health. As a result, research is continuing to develop safe natural antibiotic alternatives. In the current study, Bacillus pumilus SA388 was isolated from the chicken feces and confirmed to be a probiotic. The selected strain was tested for its antimutagenic and antioxidant capabilities before being employed as a probiotic food supplement and antibiotic alternative. The effect of B. pumilus SA388 impact on broiler chickens' growth performance, gut microbiome, blood biochemical markers, immunological response, and meat quality was also studied. B. pumilus SA388 showed significant bactericidal activity against Streptococcus pyogenes, Listeria monocytogenes, Staphylococcus aureus, Escherichia coli, Salmonella typhi, and Klebsiella pneumonia. A total of 200 chickens were used in the present study, divided equally among four experimental groups (ten birds per group with 5 replicates): group 1 (control, G1) received a basal diet without B. pumilus SA388, group 2 (G2) received a basal diet supplemented with 0.4 mg/kg of B. pumilus SA388, group 3 (G3) received a basal diet supplemented with 0.8 mg/kg of B. pumilus SA388, and group 4 (G4) received a basal diet supplemented with 1.6 mg/kg of B. pumilus SA388. Over 35 d, the B. pumilus SA388-supplemented groups outperformed the G1 in terms of body weight gain, performance index, and feed conversion ratio, with a preference for the G4 treatment. The levels of alanine aminotransferase (ALT), aspartate transaminase (AST), low-density lipoprotein (LDL), and total cholesterol decreased significantly (P < 0.05) with increasing B. pumilus SA388 dosages compared to the control G1 group. Dietary supplementation of B. pumilus SA388 at 1.6 mg/kg (G4) significantly (P < 0.05) resulted in improved lipid profile, immunological response, thyroid function, and gut microbiota compared to the control group (G1). Compared to the broilers in the control treatment (G1), the addition of B. pumilus SA388 to broilers in G4 significantly (P < 0.05) enhanced juiciness, tenderness, aroma, and taste. Adding B. pumilus SA388 to chicken feed at different doses significantly (P < 0.05) decreased average feed intake while increasing economic and relative efficiency measures. In conclusion, B. pumilus SA388 has been proven to be an effective antibiotic and nutritional supplement.

ABSTRACT Bacillus pumilus exhibits substantial economic significance, with its metabolism, adaptability, and ecological functions regulated by its bacteriophages. Here, we isolated and characterized a novel temperate phage vB_BpuM-ZY1 from B. pumilus derived from mangrove sediments by mitomycin C induction. Phage vB_BpuM-ZY1 is a typical myophage, which has an icosahedral head with a diameter of 43.34 ± 2.14 nm and a long contractible tail with a length of 238.58 ± 5.18 nm. Genomic analysis indicated that vB_BpuM-ZY1 encodes genes for lysogeny control, and its life cycle may be intricately regulated by multiple mechanisms. vB_BpuM-ZY1 was predicted to employ P2-like 5'-extended-cos packaging strategy. In addition, genome-wide phylogenetic tree and proteome tree analyses indicated that vB_BpuM-ZY1 belongs to the Peduoviridae family but forms a separate branch at a deeper taxonomic level. Particularly, the comparative genomic analysis showed that vB_BpuM-ZY1 has less than 70% intergenomic similarities with its most similar phages. Thus, we propose that vB_BpuM-ZY1 is a novel Bacillus phage belonging to a new genus under the Peduoviridae family. The protein-sharing network analysis identified 44 vB_BpuM-ZY1-related phages. Interestingly, these evolutionarily related myophages infect a broad range of hosts across different phyla, which may be explained by the high structural variations of the host recognition domain in their central spike proteins. Collectively, our study will contribute to our understanding of Bacillus phage diversity and Bacillus-phage interactions, as well as provide essential knowledge for the industrial application of B. pumilus.IMPORTANCEAlthough recent metagenomics research has obtained a wealth of phage genetic information, much of it is considered “dark matter” because of the lack of similarity with known sequences in the database. Therefore, the isolation and characterization of novel phages will help to interpret the vast unknown viral metagenome data and improve our understanding of phage diversity and phage-host interactions. Bacillus pumilus shows high economic relevance due to its wide applications in biotechnology, industry, biopharma, and environmental sectors. Since phages influence the abundance, metabolism, evolution, fitness, and ecological functions of bacteria through complex interactions, the significance of isolation and characterization of novel phages infecting B. pumilus is apparent. In this study, we isolated and characterized a B. pumilus phage belonging to a novel viral genus, which provides essential knowledge for phage biology as well as the industrial application of B. pumilus.

Cadmium (Cd) is a well-known pollutant in agricultural soil, affecting human health through the food chain. To combat this issue, Ca + Mg (25 mg L−1) nanocomposite and Bacillus pumilus, either alone or combined, were applied to rice plants under Cd (5 mg kg−1, 10 mg kg−1) contamination. In our study, growth and yield traits demonstrated the beneficial influence of Ca + Mg and B. pumilus application in improving rice defense mechanism by reducing Cd stress. Combined Ca + Mg and B. pumilus application increased SPAD (15), total chlorophyll (18), chlorophyll a (11), chlorophyll b (22), and carotenoids (21%) with Cd (10 mg kg−1), compared to the application alone. Combined Ca + Mg and B. pumilus application significantly regulated MDA (15), H2O2 (13), EL (10), and O2•– (24%) in shoots under Cd (10 mg kg−1), compared to the application alone. Cd (10 mg kg−1) increased the POD (22), SOD (21), APX (12), and CAT (13%) in shoots with combined Ca + Mg and B. pumilus application, compared to the application alone. Combined Ca + Mg and B. pumilus application significantly reduced Cd accumulation in roots (22), shoots (13), and grains (20%) under Cd (10 mg kg−1), compared to the application alone. Consequently, the combined application of Ca + Mg and B. pumilus is a sustainable solution to enhance crop production under Cd stress.

This study investigates whether Bacillus pumilus TS1 improves growth performance and alleviates inflammatory damage in broilers and explored its feasibility as an antibiotic alternative. We divided 240 one-day-old AA308 white-finned broilers into five groups (con, LPS, TS1L + LPS, TS1M + LPS and TS1H + LPS). The TS1L + LPS, TS1M + LPS and TS1H + LPS groups were fed TS1 for 15 days by gavage. The LPS, TS1L + LPS, TS1M + LPS and TS1H + LPS groups were injected intraperitoneally with 1 mg/kg LPS for three days. We investigated the probiotic and anti-inflammatory activities by measuring body weight, sequencing the intestinal flora and examining the structure of tissues by using pathological stain, real-time PCR, Western blotting and immunohistochemical detection. TS1 could improve growth performance and intestinal flora composition, also reduced different organ damage and inflammatory cytokine expression in serum and organs. The mechanism may involve upregulating HSP60 and HSP70 expression, targeting and regulating Nrf2 and P38 MAPK and modulating NF-κB and HO-1 expression at the transcriptional level in different organs. B. pumilus TS1 alleviated Inflammatory injury caused by LPS and attenuated the inflammatory response in broilers, and these effects were achieved through MAPK and Nrf2 regulation of HSPs/HO-1 in different organs. The above results suggested broilers fed with TS1 could release the LPS caused organ damage, and the most suggested dosage was 1.4 × 108 CFU/mL.

Despite their widespread applications in sectors such as pulp and paper, textile, food and beverage, pharmaceuticals, and biofuel production, laccases encounter challenges related to their activity and stability under varying reaction conditions. This review accumulates data on the complex interplay between laccase characteristics and reaction conditions for maximizing their efficacy in diverse biotechnological processes. Benefits of organic media such as improved substrate selectivity and reaction control, and their risks such as enzyme denaturation and reduced activity are reported. Additionally, the effect of reaction conditions such as pH and temperature on laccase activity and stability are gathered and reported. Sources like Bacillus pumilus, Alcaligenes faecalis, Bacillus clausii, and Bacillus tequilensis SN4 are producing laccases that are both thermo-active and alkali-active. Additionally, changes induced by the presence of various substances within reaction media such as metals, inhibitors, and organic solvents are also reported. Bacillus pumilus and Bacillus licheniformis LS04 produce the most resistant laccases in this case. Finally, the remarkable laccases have been highlighted and the proper laccase source for each industrial application is suggested. [ABSTRACT FROM AUTHOR]

Intensive poultry farming faces challenges like gut inflammation in the absence of antibiotics, resulting in reduced productivity, heightened susceptibility to enteric diseases, and other complications. Alternative strategies are needed to manage inflammation and maintain sustainable poultry production. Yaks living in highaltitude hypoxic environments have specialized gut microbes. However, yak probiotics remain largely uncharacterized. We previously isolated a strain of Bacillus pumilus (named TS2) from yaks and demonstrated its potential as a probiotic in vitro. Therefore, in this study, we evaluated the in vivo growth-promoting, antioxidant, immune, and anti-inflammatory effects of Bacillus pumilus isolated from yaks in broilers. We demonstrated the safety of TS2 isolated from yaks in broilers. Furthermore, we found that TS2 increased the average daily weight gain (ADWG) and reduced the feed conversion ratio (FCR). Supplementation with TS2 also improved the mucosal morphology, the ratio of villi to crypt cells, and enzyme activity. High-throughput sequencing showed that the abundance of Lactobacillus was higher in the TS2 treated broilers. Importantly, the serum level of malondialdehyde (MDA) was reduced and the levels of total antioxidant capacity (T-AOC) and superoxide dismutase (SOD) activity were increased in the low-dose TS2 group, while the inflammatory factors interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were downregulated compared with the control group. We demonstrated that TS2 supplementation can increase the overall growth performance and ameliorate the blood parameters related to inflammation and immunity in broilers. [ABSTRACT FROM AUTHOR]

The study aimed to develop a solid biofertilizer using Bacillus pumilus, focusing on auxin production to enhance plant drought tolerance. Methods involved immobilising B. pumilus in alginate-starch beads, focusing on microbial concentration, biopolymer types, and environmental conditions. The optimal formulation showed a diameter of 3.58 mm ± 0.18, a uniform size distribution after 15 h of drying at 30 °C, a stable bacterial concentration (1.99 × 109 CFU g−1 ± 1.03 × 109 over 180 days at room temperature), a high auxin production (748.8 µg g−1 ± 10.3 of IAA in 7 days), and a water retention capacity of 37% ± 4.07. In conclusion, this new formulation of alginate + starch + L-tryptophan + B. pumilus has the potential for use in crops due to its compelling water retention, high viability in storage at room temperature, and high auxin production, which provides commercial advantages. [ABSTRACT FROM AUTHOR]

Bacillus pumilus TUAT1 acts as plant growth–promoting rhizobacteria for various plants like rice and Arabidopsis. Under stress conditions, B. pumilus TUAT1 forms spores with a thick peptidoglycan (PGN) cell wall. Previous research showed that spores were significantly more effective than vegetative cells in enhancing plant growth. In Arabidopsis , lysin motif proteins, LYM1, LYM3 and CERK1, are required for recognizing bacterial PGNs to mediate immunity. Here, we examined the involvement of PGN receptor proteins in the plant growth promotion (PGP) effects of B. pumilus TUAT1 using Arabidopsis mutants defective in PGN receptors. Root growth of wild-type (WT), cerk1-1, lym1-1 and lym1-2 mutant plants was significantly increased by TUAT1 inoculation, but this was not the case for lym3-1 and lym3-2 mutant plants. RNA-seq analysis revealed that the expression of a number of defense-related genes was upregulated in lym3 mutant plants. These results suggested that B. pumilus TUAT1 may act to reduce the defense response, which is dependent on a functional LYM3. The expression of the defense-responsive gene, WRKY29 , was significantly induced by the elicitor flg-22, in both WT and lym3 mutant plants, while this induction was significantly reduced by treatment with B. pumilus TUAT1 and PGNs in WT, but not in lym3 mutant plants. These findings suggest that the PGNs of B. pumilus TUAT1 may be recognized by the LYM3 receptor protein, suppressing the defense response, which results in plant growth promotion in a trade-off between defense and growth. [ABSTRACT FROM AUTHOR]

The purpose of this experiment was to investigate the effects of Bacillus pumilus (GXUN613L) on the growth performance, serum biochemical indexes, antioxidant capacity, and intestinal health of coho salmon. A total of 240 coho salmon with initial body mass of (130.2±10.1) g were randomly divided into four groups with three replicates in each group and 20 coho salmon in each replicate. Bacillus pumilus (GXUN613L) was supplemented with 0, 2x107, 4x109, and 8x1011 CFU/g in basal diet in CK group, LL group, LM group, and LH group, respectively. The trial period was six weeks. The results showed that the weight gain rate and specific growth rate of coho salmon in LL group and LM group were significantly higher than those in CK group (P<0.05), the protein efficiency ratio of coho salmon in LM group was significantly higher than that in CK group (P<0.05), and the feed conversion ratio was significantly lower than that in CK group (P<0.05). Serum total protein, albumin and total cholesterol in LL group, LM group, and LH group were significantly higher than those in CK group (P<0.05). Serum glucose content and alkaline phosphatase activity in LM group were significantly higher than those in CK group (P<0.05). The total antioxidant capacity, catalase activity, superoxide dismutase, and glutathione peroxidase activity of liver in LM group were significantly higher than those in CK group (P<0.05). The muscular layer thickness and villus height of foregut and hindgut in LM group were significantly higher than those in CK group (P<0.05), and the intestinal microbial diversity was better. The result shows that the addition of Bacillus pumilus (GXUN613 L) in feed can promote the growth, improve the serum indexes and intestinal health, and enhance the antioxidant capacity of coho salmon. The suitable addition amount of GXUN613L is 4x109 CFU/g. [ABSTRACT FROM AUTHOR]

Plant growth-promoting rhizobacteria (PGPR) are biostimulants that favor plant development. We evaluated the effect of PGPR in chilli pepper (Capsicum annuum L.) and lettuce (Lactuca sativa L.). An experimental design of 5 treatments and 4 replicates was used for each crop. The bacterial species were Achromobacter xylosoxidans (C56), Arthobacter pokkalii (JLB4), Bacillus pumilus (AV5) and their combination as a consortium. The variables evaluated were: plant height, shoot diameter, relative chlorophyll index (SPAD) and freshweight of aerial part. In these experiments were observed significant differences with C56 to stimulate 11% plant height compared with the other treatments. Bacterial consortium stimulates 7.8% the relative chlorophyll index in lettuce and showing a trend to stimulate freshweight in both plants. The use of PGPR in consortium is an alternative that should be further studied to minimize the use of chemicals, enhance agricultural practicese and mitigate climate change. [ABSTRACT FROM AUTHOR]

The yak has a unique physiological structure suited to life in anoxic and cold environments at high altitudes. The aim of this study was to isolate Bacillus species with good probiotic properties from yak feces. A series of tests were performed on the isolated Bacillus: 16S rRNA identification, antibacterial activity, tolerance to gastroenteric fluid, hydrophobicity, auto-aggregation, antibiotic sensitivity, growth performance, antioxidants, and immune indexes. A safe and harmless Bacillus pumilus DX24 strain with good survival rate, hydrophobicity, auto-aggregation, and antibacterial activity was identified in the yak feces. Feeding mice with Bacillus pumilus DX24 increased their daily weight gain, jejunal villus length, villi/Crypt ratio, blood IgG levels, and jejunum sIgA levels. This study confirmed the probiotic effects of Bacillus pumilus isolated from yak feces and provides the theoretical basis for the clinical application and development of new feed additives. [ABSTRACT FROM AUTHOR]

Contamination of the environment due to speedup of anthropogenic activities has become a serious threat to modern humanity. Among the contaminants, the new emerging concern is the heavy metal (HM) contamination in the environment. Because the persistence and harmfulness of heavy metals affect the ecosystem and the health of plants, animals, and humans, they are the most toxic substances in the environment. Among them, Arsenic (As) emerged as major environmental constraint leading to enormous negative effects on the plant, animal, and human health. Even in minute quantity, As is known to cause various critical diseases in humans and toxicity in plants. Research was performed to observe the capability of plant growth–promoting strains of bacteria in enhancing Zea mays (L.) growth in arsenic polluted soil. Total 30 bacterial strains were isolated from the polluted soils, screened for plant growth promotion potential and arsenic tolerance. Eighteen isolates showed resistance to different levels of sodium arsenate (ranging from 0 to 50 mM) in agar plate using LB media. Of 18 isolates, 83.3% produced IAA, methyl red, and hydrogen cyanide; 55.5% exhibited catalase activity; 61.1% showed siderophore production; 88.8% showed phosphate solubilization; and 44.4% showed oxidase, Voges proskauer activity, and KOH solubility. The most efficient isolates SR3, SD5, and MD3 with significant arsenic tolerance and plant growth–promoting (PGP) activity were examined via sequencing of amplified 16S rRNA gene. Isolates of bacteria, i.e., SR3, SD5, and MD3, showing multiple PGP-traits were identified as Bacillus pumilus (NCBI accession number: OR459628), Paenibacillus faecalis (NCBI accession number: OR461560), and Pseudochrobactrum asaccharolyticum (NCBI accession number: OR458922), respectively. Maize seeds treated with these PGPR strains were grown in pots contaminated with 50 ppm and 100 ppm sodium arsenate. Compared to untreated arsenic stressed plants, bacterial inoculation P. asaccharolyticum (MD3) resulted 20.54%, 18.55%, 33.45%, 45.08%, and 48.55% improvement of photosynthetic pigments (carotenoid content, chlorophyll content, stomatal conductance (gs), substomatal CO2, and photosynthetic rate), respectively. Principal component analysis explained that first two components were more than 96% of the variability for each tested parameter. The results indicate that in comparison to other isolates, P. asaccharolyticum isolate can be used as efficient agent for improving maize growth under arsenic polluted soil. [ABSTRACT FROM AUTHOR]

In this study, 22 halophilic bacteria were isolated from plants collected together with rhizosphere soil from habitats with high salt content in and around Erzurum. Various plant growth-promoting (PGP) properties of these isolates (nitrogen fixing, phosphate solubilizing, ACC deaminase, and IAA and siderephore production) were determined. Bacteria positive for PGP properties and various combinations with these bacteria were subjected to pot experiments in saline medium (greenhouse conditions) and their effects on growth parameters (root and stem length, fresh and dry weight, protein, and chlorophyll and carotenoid content) of wheat plants were determined. As a result of the research, the isolates were evaluated as successful isolates 5%-41% yield increase in root length, 3.82%-54.97% in stem length, 7.14%-60.71% in wet weight, 5%-32.5% in dry weight, 13.78%-29.68% in dissolved protein content, 83.40%-120.22% in total chlorophyll content, and 45.79%-120.22% in total carotenoid content. 16S rRNA analysis showed that these 22 halophilic bacterial strains belonged to 11 different bacterial species (Bacillus pumilus, Bacillus endophyticus, Paenobacillus lautus, Planococcus citreus, Staphylococcus sciuri, Staphylococcus saprophyticus, Leucobacter iarius, Sphingomonas echinoides, Bacillus simplex, Bacillus cereus, Pantoea aggloremans). All isolates obtained in this study are new and original isolates. Inoculations with the isolates found in our study were much more effective and provided significant yield increases on wheat plants under saline irrigation compared to previous studies. These results conclude that the obtained isolates promote plant growth under saline conditions and have biofertilizer potential for wheat plants under saline conditions. [ABSTRACT FROM AUTHOR]

Herein, the potential of cotton seed hulls (CSH) as prospective feedstock for producing thermostable and alkaliphilic xylanase by Bacillus pumilus and Bacillus licheniformis during submerged fermentation was demonstrated. Results revealed that CSH predominantly contains holo-cellulose (65.6%), alpha-cellulose (38.8%), hemicelluloses (24.5%), and lignin (18.5%). Compared to the control, 0.5% CSH-supplemented growth media better supported bacterial growth (OD600 nm). Moreover, the extracellular xylanase production increased from 16 h and peaked at 24 h for both B. pumilus (359.2 U/mL) and B. licheniformis (360.8 U/mL). Bacterial xylanase demonstrated considerable stability at a higher temperature (55 °C). Likewise, significant xylanase activity was detected at pH 7.0. However, the enzyme also showed remarkable activity at alkaline pH (At pH 9.0, 84.61%, and 81.89% activity for B. pumilus and B. licheniformis, respectively), indicating alkali tolerance nature. These xylanolytic bacteria potentially depolymerized lignocellulosic fraction with a considerable release of fermentable monosaccharides (1175.10 ± 5.69 and 1278 ± 2.22 ug/mL for B. pumilus and B. lichenformis treated CSH, respectively). The scanning electron micrograph revealed the surface smoothness, cleanliness, and presence of cracks/grooves on xylanase-treated CSH surfaces. FTIR studies confirmed the stretching (at 3318 cm−1 corresponding to the O–H peak of lignin and hemicelluloses) and deformation of inter and intermolecular bonds (at 2950–2850 cm−1 related to aromatic C–H bonds present in cellulose, hemicellulose, and lignin) of lignocellulosic components of CSH. Briefly, the study highlighted the potentiality of CSH as an alternative fermentative substrate for the biosynthesis of xylanase in a biorefinery approach. [ABSTRACT FROM AUTHOR]

Drought and heat are severe environmental stresses that constantly affect plant growth and development. Maize (Zea mays L.) is known for its sensitivity to abiotic stresses, which often causes significant yield losses. With plant growth-promoting rhizobacteria (PGPR), the harmful effects of drought and heat stress on plants can be alleviated. The aim of the present study was to investigate the physiological traits of plants affected by drought stress in a treatment including four PGPR bacteria, two regimes of irrigation and two stages of phenophases, and the analysis of the influence of the individual factors and their combined effects, respectively. In addition, the investigations covered the testing of the nutrient acquisition ability and nutrient use efficiency, the plant growth stimulating effect of PGPR bacteria, and the analysis of the performance trials. The largest amount of soluble orthophosphate was produced by Pseudomonas putida (51.636 µg/cm3). This is significantly more than the amount of phosphate solubilized by Bacillus pumilus (15.601 µg/cm3), Pseudomonas fluorescens (14.817 µg/cm3) and Bacillus megaterium (14.621 µg/cm3). The germination vigour of the seedlings in the treatment with Pseudomonas putida was 55% higher, represented by the value of 15,237.125, in comparison with the control, with the value of 9792.188. The Pseudomonas putida treatment resulted in the most outstanding fresh/dry weight ratio (7.312) compared to the control (7.780), when the interaction between all factors was taken into account under stressed conditions at 50% female flowering stage. The highest dry matter content of the plants (18.344%) compared to the control (17.344%) was measured in the case of the Pseudomonas putida treatment during the milk stage of development under stressed conditions. In the field trial, the plants treated with KD2 (Bacillus pumilus and Pseudomonas putida) presented significantly higher results in the aspect of grain yield (6.278 t/ha) compared to the control (5.468 t/ha). [ABSTRACT FROM AUTHOR]

One of the problems in sea farming is infections that cause mass mortality of crustaceans. To fight infections and improve sanitary conditions, farmers are actively using probiotic preparations. We aimed to study the effect of a new probiotic based on Bacillus toyonensis B-13249 and Bacillus pumilus B-13250 strains on the incubation of Artemia franciscana cysts. Another purpose was to test a possibility of using a convolutional neural network for fast automatic counting of cysts, nauplii, and embryos. A pilot batch of the probiotic was prepared at the Prombiotech Engineering Center, Altai State University, from two strains of spore bacteria from the Center’s collection: B. toyonensis B-13249 and B. pumilus B-13250. The recommended amount of the probiotic was experimentally determined as 0.1 per 2 g of cysts. This concentration increased the number of hatched cysts by 1.4 and 10% in the batches from Lake Bolshoye Yarovoye (Z29.04) and from Lake Kuchuk (C9). It also increased the biomass yield to 7.40 ± 0.69 and 6.80 ± 0.43 g in these two batches, respectively, compared to the control samples where the yields were 5.30 ± 0.60 and 4.60 ± 0.50 g, respectively. The robot counter reduced the sample processing time 15 times and saved the data for further use. The probiotic based on B. toyonensis B-13249 and B. pumilus B-13250 had a positive effect on the hatching rate and biomass yield of A. franciscana. The new method for rapid counting of Artemia, which was based on the convolutional neural network and developed as an application of the Artemeter-1 robot, reduced the processing time and lowered labor costs.

The pharmacological importance of quinoxaline derivatives in antibacterial research is well recognized. This study focuses on the synthesis of new 2,3‐dichloroquinoxaline derivatives containing thioether/ether groups to explore their potential as potent antibacterial agents against various pathogenic bacteria. Most of the compounds exhibited significant antibacterial properties comparable to the standard drug chlorhexidine (CHX). The derivatives of 2‐chloro‐3‐(arylthiol)quinoxaline demonstrated efficacy against Escherichia coli with minimum inhibitory concentrations (MIC) of 2.5 mg/mL and minimum bactericidal concentrations (MBC) of 2.5 to 5.0 mg/mL. These derivatives also showed similar sensitivity to Bacillus pumilus. In addition, molecular docking simulations were performed to investigate the interaction between the synthesized compounds and the DNA gyrase protein (PDB ID: 1KZN), a target for antibiotics. Among the synthesized compounds, 2,3‐bis(3‐nitrophenoxy)quinoxaline exhibited the most favourable docking score of −8.36 kcal/mol, with a binding affinity comparable to that of the reference ligand clorobiocin (−9.3 kcal/mol). [ABSTRACT FROM AUTHOR]