Multiple Bacterial Pathogens Research Articles

Antibacterial potential of Thymus linearis essential oil collected from Wasturwan mountain: A combination of experimental and theoretical studies involving in silico molecular docking simulation of the major compounds against Novobiocin-resistant mutant of DNA Gyrase-B

Antibiotic resistant bacteria are immune to most antibiotics and are therefore very difficult to treat and in most cases lead to death. As such there is a pressing need for alternative and more efficient antibacterial drugs which can target these drug-resistant strains as well. The objective of this research work was to investigate the antibacterial properties of Thymus linearis essential oil (EO) against multiple disease-causing bacterial pathogens. Additionally, the study aimed to examine the molecular docking and molecular dynamic (MD) simulations of the primary components of the EO with the essential bacterial proteins and enzymes. Gas chromatography-mass spectrometry was employed to analyse the chemical composition of Thymus linearis EO. The initial screening for antibacterial properties involved the use of disc diffusion and microdilution techniques. Molecular docking studies were conducted utilising Autodock Vina. The outcomes were subsequently visualised through BIOVIA Discovery Studio. MD simulations were conducted using iMODS, an internet-based platform designed for MD simulations. The essential oil (EO) was found to contain 26 components, with thymol, carvacrol, p-cymene, and γ-terpinene being the primary constituents. The study findings revealed that Thymus linearis EO demonstrated antibacterial effects that were dependent on both the dose and time. The results of molecular docking studies revealed that the primary constituents of the EO, namely thymol, carvacrol, and p-cymene, exhibited robust interactions with the active site of the bacterial DNA gyrase enzyme. This finding provides an explanation for the antibacterial mechanism of the EO. The results indicate that Thymus linearis EO possesses potent antibacterial properties against the MDR microorganisms. Molecular docking analyses revealed that the essential oil's primary components interact with the amino acid residues of the DNA-Gyrase B enzyme, resulting in a favourable docking score.

Multiple infections of zoonotic pathogens in wild Brandt's voles (Lasiopodomys brandtii).

The frequent interactions of rodents with humans make them a common source of zoonotic infections. Brandt's vole is the dominant rodent species of the typical steppe in Inner Mongolia, and it is also an important pest in grassland. To obtain an initial unbiased measure of the microbial diversity and abundance in the blood and intestinal tracts and to detect the pathogens carried by wild Brandt's voles in Hulun Buir, Inner Mongolia. Twenty wild adult Brandt's voles were trapped using live cages, and 12 intestinal samples were collected for metagenomic analysis and 8 blood samples were collected for meta-transcriptomic analysis. We compared the sequencing data with pathogenic microbiota databases to analyse the phylogenetic characteristics of zoonotic pathogens carried by wild voles. A total of 122 phyla, 79 classes, 168 orders, 382 families and 1693 genera of bacteria and a total of 32 families of DNA and RNA viruses in Brandt's voles were characterized. We found that each sample carried more than 10 pathogens, whereas some pathogens that were low in abundance were still at risk of transmission to humans. This study improves our understanding of the viral and bacterial diversity in wild Brandt's voles and highlights the multiple viral and bacterial pathogens carried by this rodent. These findings may serve as a basis for developing strategies targeting rodent population control in Hulun Buir and provide a better approach to the surveillance of pathogenic microorganisms in wildlife.

Integration of microbiome and Koch’s postulates to reveal multiple bacterial pathogens of whitish muscle syndrome in mud crab, Scylla paramamosain

BackgroundFor more than a century, the Koch’s postulates have been the golden rule for determining the causative agents in diseases. However, in cases of multiple pathogens-one disease, in which different pathogens can cause the same disease, the selection of microorganisms that regress infection is hard when Koch’s postulates are applied. Microbiome approaches can obtain relatively complete information about disease-related microorganisms and can guide the selection of target microorganisms for regression infection. In the present study, whitish muscle syndrome (WMS) of Scylla paramamosain, which has typical symptoms with whitish muscle and blackened hemolymph was used as an example to establish a new research strategy that integrates microbiome approaches and Koch’s postulates to determinate causative agents of multiple pathogens-one disease.ResultsMicrobiome results revealed that Aeromonas, Acinetobacter, Shewanella, Chryseomicrobium, Exiguobacterium, Vibrio and Flavobacterium, and Kurtzmaniella in hemolymph were bacterial and fungal indicators for WMS. A total of 23 bacteria and 14 fungi were isolated from hemolymph and muscle tissues, and among the bacteria, Shewanella chilikensis, S. xiamenensis, Vibrio alginolyticus, S. putrefaciens, V. fluvialis, and V. parahaemolyticus were present in hemolymph and/or muscle tissues in each WMS crab, and the last three species were also present in three Healthy crabs. The target bacteria and fungi were further screened to regression infections based on two criteria: whether they belonged to the indicator genera for WMS, whether they were isolated from both hemolymph and muscle tissues in most WMS crabs. Only S. chilikensis, S. putrefaciens, S. xiamenensis, V. alginolyticus, V. fluvialis, and V. parahaemolyticus met both two criteria. The six bacteria that met both two criteria and six fungi and another bacterium that unmatched any of two criteria were used to perform regression infection experiments based on Koch’s postulates. S. chilikensis, S. putrefaciens, S. xiamenensis, V. alginolyticus, V. fluvialis, and V. parahaemolyticus met both two criteria, and the results indicate that they cause WMS in crabs independently.ConclusionsThis study fully demonstrated that our research strategy that integrates the microbiome and Koch’s postulates can maximize the ability to catch pathogens in one net for the situation of multiple pathogens-one disease.EgKJnBsvf5xgzELdpRDt_QVideo

Microhabitat modeling of the invasive Asian longhorned tick (Haemaphysalis longicornis) in New Jersey, USA

The Asian longhorned tick (Haemaphysalis longicornis) is a vector of multiple arboviral and bacterial pathogens in its native East Asia and expanded distribution in Australasia. This species has both bisexual and parthenogenetic populations that can reach high population densities under favorable conditions. Established populations of parthenogenetic H. longicornis were detected in the eastern United States in 2017 and the possible range of this species at the continental level (North America) based on climatic conditions has been modeled. However, little is known about factors influencing the distribution of H. longicornis at geographic scales relevant to local surveillance and control. To examine the importance of local physiogeographic conditions such as geology, soil characteristics, and land cover on the distribution of H. longicornis we employed ecological niche modeling using three machine learning algorithms - Maxent, Random Forest (RF), and Generalized Boosting Method (GBM) to estimate probability of finding H. longicornis in a particular location in New Jersey (USA), based on environmental predictors. The presence of H. longicornis in New Jersey was positively associated with Piedmont physiogeographic province and two soil types - Alfisols and Inceptisols. Soil hydraulic conductivity was the most important predictor explaining H. longicornis habitat suitability, with more permeable sandy soils with higher hydraulic conductivity being less suitable than clay or loam soils. The models were projected over the state of New Jersey creating a probabilistic map of H. longicornis habitat suitability at a high spatial resolution of 90×90 meters. The model's sensitivity was 87% for locations sampled in 2017-2019 adding to the growing evidence of the importance of soil characteristics to the survival of ticks. For the 2020-2022 dataset the model fit was 57%, suggestive of spillover to less optimal habitats or, alternatively, heterogeneity in soil characteristics at the edges of broad physiographic zones. Further modeling should incorporate abundance and life-stage information as well as detailed characterization of the soil at collection sites. Once critical parameters that drive the survival and abundance of H. longicornis are identified they can be used to guide surveillance and control strategies for this invasive species.

Inferring the heritability of bacterial traits in the era of machine learning.

The R codes and data used in the numerical experiments are available at: https://github.com/tienmt/her_MLs.

Combined Effect of Isolated Bacteriophage and Neem Extract on Isolated Multiple Drug-Resistant Pathogenic Escherichia coli E1 From Well Water.

Multiple drug-resistant Escherichia coli (E. coli) is a serious cause of concern, and they can be observed in hospital settings, natural environment, and animals. Dissemination of multiple drug-resistant (MDR) E. coli can pose a high risk to public health. Moreover, they are hard to control with commercial antibiotics, since they have acquired resistance against most of them. Therefore, to control multiple drug-resistant bacteria, alternative strategies have been adopted such as phage therapy, herbal remedies, nanoparticles etc. In the current study, the combined application of neem leaf extract and bacteriophage is used to control an isolated multiple drug-resistant E. coli E1. We have applied 0.1 mg/ml concentration of neem extract in combination with an isolated phage vB_EcoM_C2 of 1011 titer and found that the combinatorial treatment approach significantly controls the growth of E. coli E1 as compared to a single non-combinatorial treatment. In this study, every E. coli cell is targeted by 2 antimicrobials (phage and neem extract) at the same time, which is more effective as compared to the sole treatment. Implementation of the neem extract with phage opens a new alternative approach to the chemotherapeutics for the control of multiple drug-resistant bacterial pathogens. This approach may be effective, economical, and eco-friendly to combat MDR.

Design, synthesis and antibacterial activity of a class of novel molecular engineering analogues of Tachyplesin I

Tachyplesin I (TP-I) is a marine-derived antimicrobial peptide isolated from the hemocyte membrane of Japanese horseshoe crab ( Tachypleus tridentatus ) with broad antimicrobial spectrum. It consists of 17 amino acid residues and shows a characteristic structure with three tandem repeats of a tetrapeptide sequence. The three tandem repeats of a tetrapeptide sequence are the key components of the amphipathic β-hairpin structure of TP-I and we proposed that these fragments were critical for the antimicrobial activity, and designed a class of novel molecular engineering analogs of TP-I via re-combination of the three tetrapeptide fragments with different numbers of disulfide bonds in a search of therapeutically valuable lead compound. The synthesis of cyclic peptides with 1/2 disulfide bond(s) are challenging and we successfully prepared most of them using the orthogonal Cys protection strategy. The antimicrobial assay result revealed that the molecular engineering analogs of TP-I via fragment re-combination with 2 disulfide bonds maintained the antimicrobial potency across the range of microbes and confirmed the importance of tetrapeptide fragments and disulfide bonds. Of particular interest, analogue TP-I-R2 exhibited good antimicrobial activity against most of the tested microbes with MIC ranging from 4 to 128 μg/mL. Moreover, TP-I-R1, TP-I-R1b and TP-I-R3 particularly exhibited higher potency of more than 16/16/8-fold against Grampositive Bacillus pumilus strains than TP-I, TP-I-R2 and TP-I-R4 showed comparable potency against Gramnegtive Salmonella typhimurium strains with TP-I. These compounds may be considered as promising lead for further development toward new treatments for infections caused by multiple bacterial pathogens. • A class of novel molecular engineering analogues of Tachyplesin I (TP-I) were designed and synthesized. • TP-I-R2 exhibited broader antimicrobial spectrum and good antimicrobial potency. • TP-I-R2 could be considered as promising lead for the dicovery of novel antibiotics.

Microbial lectome versus host glycolipidome: How pathogens exploit glycosphingolipids to invade, dupe or kill.

Glycosphingolipids (GSLs) are ubiquitous components of the cell membranes, found across several kingdoms of life, from bacteria to mammals, including humans. GSLs are a subclass of major glycolipids occurring in animal lipid membranes in clusters named “lipid rafts.” The most crucial functions of GSLs include signal transduction and regulation as well as participation in cell proliferation. Despite the mainstream view that pathogens rely on protein–protein interactions to survive and thrive in their hosts, many also target the host lipids. In particular, multiple pathogens produce adhesion molecules or toxins that bind GSLs. Attachment of pathogens to cell surface receptors is the initial step in infections. Many mammalian pathogens have evolved to recognize GSL-derived receptors. Animal glycosphingolipidomes consist of multiple types of GSLs differing in terminal glycan and ceramide structures in a cell or tissue-specific manner. Interspecies differences in GSLs dictate host specificity as well as cell and tissue tropisms. Evolutionary pressure exerted by pathogens on their hosts drives changes in cell surface glycoconjugates, including GSLs, and has produced a vast number of molecules and interaction mechanisms. Despite that abundance, the role of GSLs as pathogen receptors has been largely overlooked or only cursorily discussed. In this review, we take a closer look at GSLs and their role in the recognition, cellular entry, and toxicity of multiple bacterial, viral and fungal pathogens.

BIRC6 modifies risk of invasive bacterial infection in Kenyan children.

Invasive bacterial disease is a major cause of morbidity and mortality in African children. Despite being caused by diverse pathogens, children with sepsis are clinically indistinguishable from one another. In spite of this, most genetic susceptibility loci for invasive infection that have been discovered to date are pathogen specific and are not therefore suggestive of a shared genetic architecture of bacterial sepsis. Here, we utilise probabilistic diagnostic models to identify children with a high probability of invasive bacterial disease among critically unwell Kenyan children with Plasmodium falciparum parasitaemia. We construct a joint dataset including 1445 bacteraemia cases and 1143 severe malaria cases, and population controls, among critically unwell Kenyan children that have previously been genotyped for human genetic variation. Using these data, we perform a cross-trait genome-wide association study of invasive bacterial infection, weighting cases according to their probability of bacterial disease. In doing so, we identify and validate a novel risk locus for invasive infection secondary to multiple bacterial pathogens, that has no apparent effect on malaria risk. The locus identified modifies splicing of BIRC6 in stimulated monocytes, implicating regulation of apoptosis and autophagy in the pathogenesis of sepsis in Kenyan children.

Bacteriophages: A possible solution to combat enteropathogenic Escherichia coli infections in neonatal goats.

Due to awareness and benefits of goat rearing in developing economies, goats' significance is increasing. Unfortunately, these ruminants are threatened via multiple bacterial pathogens such as enteropathogenic Escherichia coli (EPEC). In goat kids and lambs, EPEC causes gastrointestinal disease leading to substantial economic losses for farmers and may also pose a threat to public health via the spread of zoonotic diseases. Management of infection is primarily based on antibiotics, but the need for new therapeutic measures as an alternative to antibiotics is becoming vital because of the advent of antimicrobial resistance (AMR). The prevalence of EPEC was established using bfpA gene, uspA gene and Stx1 gene, followed by phylogenetic analysis using Stx1 gene. The lytic activity of the isolated putative coliphages was tested on multi-drug resistant strains of EPEC. It was observed that a PCR based approach is more effective and rapid as compared to phenotypic tests of Escherichia coli virulence. It was also established that the isolated bacteriophages exhibited potent antibacterial efficacy invitro, with some of the isolates (16%) detected as T4 and T4-like phages based on gp23 gene. Hence, bacteriophages as therapeutic agents may be explored as an alternative to antibiotics in managing public, livestock and environmental health in this era of AMR.

Rate of Multiple Viral and Bacterial CoInfection(s) in Influenza A/H9N2-Infected Broiler Flocks.

Repeated cases of low pathogenic influenza A/H9N2 virus (IAV/H9N2) have been reported in commercial chickens since its emergence in 1998 in Pakistan. However, recently increased mortality and severe respiratory complications under field conditions have been noticed, suggesting concomitant influenza infections with respiratory viral and/or bacterial pathogens. Therefore, the present study aimed to investigate the presence of IAV/H9N2 coinfecting with multiple viral and bacterial pathogens in broiler chicken flocks. We surveyed 60 broiler flocks with respiratory signs from March through July 2019 in Punjab, Pakistan. Suspected flocks were screened for the presence of IAV using a lateral-flow device. Tracheal, cloacal, and bone marrow samples were collected and further tested for seven viral agents (chicken anemia; Newcastle disease; infectious bronchitis; infectious laryngeotracheitis [ILT]; and IAV subtypes H9, H7, and H5) and three bacterial agents (Mycoplasma gallisepticum; Mycoplasma synovae; Ornithobacterium rhinotracheale [ORT]) using PCR assays. Upon initial screening for IAV, 35/60 (58.3%) flocks tested positive. The coinfection of IAV/H9N2 with other pathogens was detected in 25 (71.4%) flocks and only IAV/H9N2 was detected in 10 (28.6%) flocks out of total positive IAV flocks (n = 35). IAV subtypes H5 and H7, ILT, and ORT were not detected throughout the study period. The detection rate of double, triple, and quadruple combinations of coinfections with IAV/H9N2 were 37% (13 flocks), 26% (9 flocks), 9% (3 flocks), respectively. Higher average mortality (28.5%) was found in broiler chicken flocks coinfected with viral and/or bacterial pathogens than in flocks where only H9 low pathogenic IAV/H9N2 was detected (20.8%). In conclusion, higher circulation of IAV/H9N2 with other viral and bacterial pathogens may contribute to higher production and economic losses at the farm level.

Listeria monocytogenes requires cellular respiration for NAD+ regeneration and pathogenesis.

Cellular respiration is essential for multiple bacterial pathogens and a validated antibiotic target. In addition to driving oxidative phosphorylation, bacterial respiration has a variety of ancillary functions that obscure its contribution to pathogenesis. We find here that the intracellular pathogen Listeria monocytogenes encodes two respiratory pathways which are partially functionally redundant and indispensable for pathogenesis. Loss of respiration decreased NAD+ regeneration, but this could be specifically reversed by heterologous expression of a water-forming NADH oxidase (NOX). NOX expression fully rescued intracellular growth defects and increased L. monocytogenes loads >1000-fold in a mouse infection model. Consistent with NAD+ regeneration maintaining L. monocytogenes viability and enabling immune evasion, a respiration-deficient strain exhibited elevated bacteriolysis within the host cytosol and NOX expression rescued this phenotype. These studies show that NAD+ regeneration represents a major role of L. monocytogenes respiration and highlight the nuanced relationship between bacterial metabolism, physiology, and pathogenesis.

In silico and in vivo approaches to recombinant multi-epitope immunogen of GroEL provides efficient cross protection against S. Typhimurium, S. flexneri, and S. dysenteriae

ObjectivesStress or Heat Shock Proteins (HSPs) have been included in various operations like protein folding, autophagy, and apoptosis. HSP families recognize as protective antigens in a wide range of bacteria because they have been conserved through evolution. Due to their homology as well as antigenicity they are competent for applying in cross-protection against bacterial diseases. MethodsIn the present study, bioinformatics approaches utilized to design epitope-based construction of Hsp60 (or GroEL) protein. In this regard, potential B-cell and T-cell epitopes except for allergenic sequences were selected by immunoinformatic tools. The structural and functional aspects of the DNA, RNA, and protein levels were assessed by bioinformatics software. Following in silico investigations, recombinant GroEL multi-epitope of Salmonella typhi was expressed, purified, and validated. Mouse groups were immunized with recombinant protein and humoral immune response was measured by enzyme linked immunosorbent assay (ELISA). Animal challenge against Salmonella Typhimurium, Shigella flexneri, and Shigella dysenteriae was evaluated. Resultsrecombinant protein expression and purification with 14.3 kilodaltons (kDa) was confirmed by SDS-PAGE and western blotting. After animal administration, the immunoglobulins evaluated increase after each immunization. Immunized antisera exhibited 80%, 40%, and 40% protection against the lethal dose infection by S. Typhimurium, S. flexneri, and S. dysenteriae respectively. Passive immunization conferred 50%, 30%, and 30% protection in mice against S. Typhimurium, S. flexneri and S. dysentery respectively. In addition, bacterial organ load had exhibited a significant decrease in colony forming unit (CFU) in the liver and spleen of the immunized mice compared to the control. ConclusionOur study demonstrates the efficacy of S. Typhi recombinant GroEL multi-epitope to consider as a universal immunogen candidate versus multiple bacterial pathogens.

Seroprevalence and associated factors of viral agents of the bovine respiratory disease complex in buffaloes of Colombia

ABSTRACT: The bovine respiratory disease complex causes sizable losses for livestock producers. Its presentation involves the interaction among multiple viral and bacterial pathogens. Viral agents include: bovine viral diarrhea virus (BVDV), bovine alphaherpesvírus 1 (BoHV-1), bovine parainfluenza virus (bPI3v) and bovine respiratory syncytial virus (BRSV); so far, these viruses have been little studied in the buffalo population. This study is aimed to assess the presence of these viruses in buffalo populations of the department of Córdoba, Colombia. A transversal study was conducted upon assessment of 37 farms and 861 buffaloes. Seroprevalence in animals were as follows: 21.7% (BVDV), 51.5% (BoHV-1), 73.6% (bPI3v) and 58.9% (BSRV), while seroprevalence in farms stood at: 94.6% (BVDV), 100% (BoHV-1), 100% (bPI3v) and 100% (BSRV). Conversely, the multiple seropositivity in buffaloes or past coinfection of viral agents amounted to 23.3% for a single virus, 31.8% for two viruses, 29.4% for three viruses and 7.1% for four viruses; only 8.4% of buffaloes were seronegative for four viruses. This is the first regional epidemiological study in the buffalo populations of Colombia and the results showed viral circulation and multiple seropositivity in animals; in addition, the high prevalence found pose a concerning epidemiological threat.

Synthesis, characterization, and antimicrobial activity of lipophilic N,N′‐bis‐substituted triazolium salts

AbstractA series of N,N′‐bis‐substituted triazolium salts have been synthesized using 1H‐1,2,4‐triazole, 1H‐1,2,3‐triazole, and 1H‐1,2,3‐benzotriazole. Synthesized compounds were tested for their antimicrobial activities against a panel of representative ESKAPE pathogens, which are common culprits of hospital‐acquired bacterial infections. Compounds 7 and 12 displayed significant antimicrobial activities against all pathogens including a vancomycin‐resistant strain of Enterococcus faecium and a multidrug resistant strain of Acinetobacter baumannii. Furthermore, the structure activity relationship of N,N′‐bis‐substituted triazolium salts revealed a potential selectivity of gram‐positive or gram‐negative bacteria based on the heterocycle center. Thus, N,N′‐bis‐substituted triazolium salts exhibit potent antimicrobial properties against multiple ESKAPE bacterial pathogens which warrants a further investigation of their biological activities.

Characterization of bacteriocin and chitinase producing bacterial isolates with broad-spectrum antimicrobial activities

There is a need for more efficient and eco-friendly approaches to overcome increasing microbial infections. Bacteriocins and chitinases from Bacillus spp. can be powerful alternatives to conventional antibiotics and antifungal drugs, respectively. The purpose of this study was to assess the inhibitory potential of bacteriocins and chitinase enzymes against multiple resistant bacterial and fungal pathogens. Bacterial isolates were selected by growth on minimal salts medium and after that were morphologically and biochemically characterized. The physiochemical characterization of bacteriocins was carried out. The inhibitory potential of bacteriocins towards six pathogenic bacteria was determined by the well diffusion assay while chitinase activity towards three fungal strains was determined by the dual plate culture assay. Two bacterial strains (WW2P1 and WRE4P2), out of nine showed inhibition of K. pneumonia, P. aeruginosa, E. coli and MRSA while WW4P2 was positive against S. typhimurium and E. coli and WRE10P2 against P. aeruginosa, S. pneumoniae. Two bacterial isolates (WW3P1 and WRE10P2) were chosen for further study on the basis of their antifungal activities. Of these, WW3P1 isolate was more effective against A. fumigatus as well as A. niger. The proteinaceous nature of the bacteriocins was confirmed by treatment of the crude extract with proteinase K. It was found that the inhibitory activity of strain WW3P1 against E. coli was highest at 20 °C, and against S. pneumoniae it was at 20 °C and pH 10 after treatment with EDTA. Inhibition by strain the WRE10P2 against P. aeruginosa was highest at 20 °C and pH 14. It was found that EDTA increased the inhibitory activity of strain WW2P1 against P. aeruginosa, K. pneumoniae and E. coli by 2 ± 0.235, 3.5 ± 0.288, 2.5 ± 1.040 times, respectively, of strain WRE4P2 against P. aeruginosa and E. coli by 2.5 ± 0.763, 2.7 ± 0.5 times, respectively, and of strain WRE10P2 against S. pneumoniae by 3 ± 0.6236 times. The isolates have promising inhibitory activity, which should be further analyzed for the commercial production of antimicrobials.Article highlightsThe current study aimed to isolate the microbiome from wheat plant (Triticum aestivum L.), to screen for bacteriocin production and to assess its antimicrobial activity against human pathogens.Forty-one phenotypically different bacterial colonies were subjected to bacteriocin purification from which 25 colonies showed positive reactions.These 25 bacterial isolates were screened against six different human bacterial pathogens using the well diffusion method to check the antimicrobial activity.Out of nine bacterial isolates, WW3P1 and WRE10P2 were able to degrade the chitin and utilize it as their sole energy source. Strain WRE4P2 exhibited partial inactivation in its activity against MRSA after treatment with proteinase K.

Screening of Antagonistic Bacteria from Endophytes against Walnut Blight Pathogen Xanthomonas arboricola pv. juglandis

Walnut blight caused by Xanthomonas arboricola pv. juglandis (Xaj) is the most important bacterial disease in walnut production worldwide. To seek biocontrol agents against Xaj, we screened 152 endophytic bacteria isolated from 87 plants. Through dual-culture method screening, we obtained four antagonistic bacteria, ATE17, BME17, CIE17, and OFE17 which were isolated from Amaranthus tricolor, Bambusa multiplex, Canna indica, and Osmanthus fragrans plants respectively. The inhibition ratios of ATE18, BME17, CIE18, and OFE17 against Xaj on plates were 1.5, 1.6, 1.3, and 1.6, respectively. These indicated they have good biocontrol potential for walnut bacterial blight. Subsequently, the four endophytic bacteria were identified by morphology, Gram staining, Microbial Identification System (fatty acid methyl ester analysis), as well as 16S rDNA and gyrB sequencing. It turns out that all four strains were identified as Bacillus sp. Furthermore, the two strains BME17 and OFE17 can suppress multiple plant fungal pathogens and bacterial pathogens on plates.

Antimicrobial efficacy and activity of ethanolic extract of Piper betle L. on Staphylococcus aureus-infected wound in mice and clinical isolates of multiple drug-resistant bacterial pathogens.

This study aimed to determine the in vivo effectiveness of the ethanolic extract of Piper betle L. leaves against Staphylococcus aureus-infected wounds in mice and its antimicrobial properties on clinical isolates of multiple drug-resistant bacterial pathogens. Twenty mice were divided into four groups. Wounds were created in all mice under anesthesia by excision from the dorsal skin down to the subcutaneous fat and inoculating with S. aureus. After 24 h, the wound of each mouse was treated once daily by application of the respective cream. Group I was treated with mupirocin antibacterial cream; Group II received a cream base containing no active ingredient; Groups III and IV were treated with 2.5% and 5.0% concentrations of P. betle cream, respectively. Further, an in vitro study was performed by adding undiluted, 1:50 and 1:100 dilutions of the four studied creams in normal saline containing 1.5 × 108 CFU/mL of the following bacteria: antimicrobial-susceptible S. aureus, Escherichia coli, Pseudomonas aeruginosa, methicillin-resistant S. aureus, extended-spectrum β-lactamase-producing Escherichia coli, vancomycin-resistant Enterococcus, metallo-βlactamase-producing P. aeruginosa and carbapenem-resistant Klebsiella pneumoniae. The mice in Groups III and IV had significantly faster wound contraction and significantly shorter reepithelialization time than Group II (p < 0.05), which were not significantly different from Group I (p > 0.05). P. betle creams inhibited all studied bacterial strains at full concentration and at a dilution of 1:50. The inhibitory effect was more significant than Groups I and II (p < 0.05), except on S. aureus. Specifically, S. aureus inhibition was not significantly different for Groups III and IV (p > 0.05) when compared with Group I. Cream formulations derived from P. betle ethanolic extract have great potential as antimicrobial agents for the treatment of wound infection. Further clinical tests are recommended to determine the safety and efficacy of these formulations in other mammalian species.

Isolation and Identification of Multiple Drug Resistant Bacterial Pathogens from Well Water Samples in and Around Wolaita Sodo Town, Southern Ethiopia

Water for human consumption is required to be free from any bacteria that might pose a health risk. The worldwide prevalence of pathogen contamination is a serious concern, and enhancing the understanding of major pathogen sources and their significant impacts on water resources is crucial. Further, these organisms underwent multiple drug resistance in their systems. The present study mainly focused on Multiple Drug Resistant (MDR) bacterial pathogens in well water system in and around Wolaita Sodo Town. A total of 380 well water samples were collected and screened for SPC, enrichment process and selective media isolation. Further all the isolates were used to know the sensitivity/resistant patterns by Bauer-Kirby method and based on resistant pattern MDR isolates were assessed and recorded. Amon 380 well water samples screened for water potability by SPC method, 154 samples were positive. Among 154 SPC positive water samples yielded 106 different isolates of bacteria and it includes five genera, namely Escherichia coli, Salmonella spp., Shigella spp., Proteus spp. and Enterococcus spp. All the isolated five genus were confirmed by preliminary characters, colony morphology and biochemical tests. Among 106 isolates, 13 isolates of E. coli, 3 isolates of Salmonella spp., 21 isolates of Shigella spp., 9 isolates of Proteus spp., and 4 isolates of Enterococcus spp. were documented as MDR bacterial pathogens. The high prevalence rate of multiple drug resistant bacterial pathogens in the well water samples could potentially pose a threat to people consuming this water. Therefore, the present study suggesting to monitor the quality of water and strict quality control measures should be put in place to ensure the effective treatment of drinking water.&#x0D; Keywords: Water samples, Standard Plate Count, Antibacterial Susceptibility Test and MDR

S100A9 modulates neutrophil extracellular trap formation thereby enhancing macrophage killing of bacterial pathogens.

Abstract Professional phagocytes form the backbone of the innate immune response and are equipped with an arsenal of antimicrobial processes. While many of these processes have been explored in isolation, how immune cells cooperatively combat bacterial pathogens at the site of infection is unclear. The proteins forming calprotectin, S100A8 and S100A9, are the most abundant cytosolic proteins in neutrophils and important components of the innate immune response. Surprisingly, S100A9-deficient (A9−/−) mice are protected from Staphylococcus aureus infection with lower bacterial burdens in the heart. We have uncovered an intracellular function for S100A9, where A9−/− neutrophils produce more mitochondrial superoxide, thereby promoting the terminal formation of neutrophil extracellular traps (suicidal NETosis). Increased suicidal NETosis does not heighten neutrophil killing of S. aureus in isolation, but rather augments macrophage (Mϕ) killing. More specifically, NET formation enhances Mϕ antibacterial activity coinciding with Mϕ phagocytosis of S. aureus and transferring neutrophil-specific antimicrobial peptides to Mϕs. Finally, similar results are observed in response to other phylogenetically distinct bacterial pathogens implicating this as an immune defense mechanism that broadly enhances antibacterial activity. Based on these results, we propose that intracellular S100A9 is a critical rheostat for neutrophil function during infection. Further, NET formation acts as a conduit for neutrophils to enhance the antibacterial activity of Mϕs, where accelerated suicidal NETosis by A9−/− neutrophils makes them especially adept at increasing the antibacterial activity of Mϕs against multiple bacterial pathogens.