Providencia Alcalifaciens Research Articles

Innate immune Galectin-7 specifically targets microbes that decorate themselves in blood group-like antigens

SummaryAdaptive immunity can target a nearly infinite range of antigens, yet it is tempered by tolerogenic mechanisms that limit autoimmunity. Such immunological tolerance, however, creates a gap in adaptive immunity against microbes decorated with self-like antigens as a form of molecular mimicry. Our results demonstrate that the innate immune lectin galectin-7 (Gal-7) binds a variety of distinct microbes, all of which share features of blood group-like antigens. Gal-7 binding to each blood group expressing microbe, including strains of Escherichia coli, Klebsiella pneumoniae, Providencia alcalifaciens, and Streptococcus pneumoniae, results in loss of microbial viability. Although Gal-7 also binds red blood cells (RBCs), this interaction does not alter RBC membrane integrity. These results demonstrate that Gal-7 recognizes a diverse range of microbes, each of which use molecular mimicry while failing to induce host cell injury, and thus may provide an innate form of immunity against molecular mimicry.

ARGs Prevalence Among Hospitalized COVID‐19 Patients with Bacterial Infections and Resistome abundance

BackgroundThe burden of antimicrobial resistance AMR worldwide is substantial and is growing. Many factors play a role in the emergence of resistance. Our microbiome is a significant reservoir for antimicrobial resistance genes (ARGs). The use and misuse of antimicrobials can select multi‐resistant bacteria and modify the repertoire of ARGs in the gut microbiota. There is clear evidence that hospitalized COVID patients have altered gut microbiota that might affect the prevalence and abundance of AMR. The World Health Organization (WHO) has reported 364 million cases worldwide. Moreover, the CDC announced 72 million positives with more than 4.2 million hospitalized cases in the US alone.ObjectiveThe purpose of the current study is to review AMGs prevalence among hospitalized COVID‐19 patients with bacterial infections and explore the alteration of resistome abundance.MethodologyWe searched the pathogen detection of the National Center for Biotechnology database for hospitalized positive COVID‐19 cases with microbiome sequencing submission using the following keywords: antimicrobial resistance, antibiotic stewardship, prevalence, epidemiology, mechanism of resistance, and COVID‐19, SARS‐CoV‐2, bacterial infection, hospitalization, healthcare‐associated infection, antibiotic resistance, antimicrobial resistance, multi‐drug resistance, Acinetobacter, Klebsiella, Streptococcus, Staphylococcus, Pseudomonas, Escherichia, Enterococcus, Methicillin‐resistant Staphylococcus aureus, New Delhi Metallo‐β‐lactamase‐producing carbapenem‐resistant enterobacterales, Severe acute respiratory syndrome coronavirus 2, Vancomycin‐resistant enterococcus. Web‐based pipelines ResistoXplorer and Qiime2 and different databases, including RESfinder, Card, and ARGminer, are used for the metagenomics analysis. They are mainly based on programming languages ‐ JavaScript (version: 1.2.0) and R (version: 4.1.0).ResultsOur preliminary results showed that, to date, 11,129 isolates were detected worldwide between the declaration of the covid as a pandemic and January 2022. Moreover, the isolates prevalence was Acinetobacter baumannii at 39.7%, followed by Klebsiella pneumonia at 32.2%, then E.coli and Shigella at 11.7%, Enterobacter at 5.4%, and Pseudomonas aeruginosa 2.5%. Finally, infections such as Klebsiella oxytoca, Citrobacter freundii, and Providencia alcalifaciens were around 10%. We explored the genetic diversity of MRGs, including: aac(6′)‐Ib′, aadA1, and(3′′)‐IIa, aph(3′′)‐Ib, aph(3′)‐Ia, aph(6)‐Id, armA, arr‐2, blaADC‐30, blaOXA‐23, blaOXA‐66, blaOXA. Currently, we are working on WGS and 16s sequencing analysis, and our future work includes data analysis on abundance profiles and exploring the resistome signatures generated from AMR metagenomics of the microbiota of positive COVID‐ hospitalized patients.ConclusionWe are anticipating that there will be a significant change take place in the gut microbiome, hence the ARGs and resistome abundance in patients with COVID‐19.

Providencia alcalifaciens is a Highly Antimicrobial Resistant Bacteria Found in a Suburban Creek

Antibiotic resistant bacteria have been found in diverse environments all over the world with greater rates of resistance in those locations most closely in contact with human populations. The Navajo Creek is a small, suburban waterway 15 miles southwest of Chicago, IL that flows through neighborhoods and forest preserves. Among bacteria isolated from the creek, resistance to five different antibiotics has been discovered, and metagenomic analysis of these populations shows diversity among the resistant microbes. Strikingly, one bacterial species, Providencia alcalifaciens,shows resistance to all five antibiotics simultaneously. Full genome analysis reveals that this species displays a diverse set of resistance genes to overcome these drugs. The interplay between these genes and the drugs they defeat is still being determined. Additionally, the full range of P. alcalifaciensin the local environment is being measured by identifying the microbe in regional water, soil, and animal waste. This project offers careful analysis of a highly resistant microbe and potential pathogen that resides in a suburban waterway.

Analysis of Antibiotic Resistance and Biofilm-Forming Capacity in Tetracycline-Resistant Bacteria from a Coastal Lagoon

Concerns have been raised regarding co-selection for antibiotic resistance among bacteria exposed to antibiotics used as growth promoters for some livestock and poultry species. Tetracycline had been commonly used for this purpose worldwide, and its residue has been associated with selection of resistant bacteria in aquatic biofilms. This study aimed to determine the resistance profile, the existence of some beta-lactamases genes and the capacity to form biofilm of bacteria isolated from water samples previously exposed to tetracycline (20 mg/L). Thirty-seven tetracycline-resistant bacterial strains were identified as Serratia marcescens, Escherichia coli, Morganella morganii, Pseudomonas aeruginosa, Citrobacter freundii, Providencia alcalifaciens, and Enterococcus faecium. The highest percentage of resistance was for ampicillin (75.75%) and amoxicillin/clavulanic acid (66.66%) in the Gram-negative bacteria and an E. faecium strain showed high resistance to vancomycin (minimum inhibitory concentration 250 μg/mL). Among the strains analyzed, 81.09% had multidrug resistance and eight Gram-negatives carried the blaOXA-48 gene. All strains were able to form biofilm and 43.23% were strong biofilm formers. This study suggests that resistant bacteria can be selected under selection pressure of tetracycline, and that these bacteria could contribute to the maintenance and spread of antimicrobial resistance in this environment.

Closed Genome Sequences of Providencia alcalifaciens Isolates from Dogs.

ABSTRACTEight Providencia alcalifaciens isolates from eight different dogs in Norway with acute hemorrhagic diarrhea were sequenced. Based on Illumina and Oxford Nanopore Technologies sequencing, all of the genomes were complete and closed after hybrid assembly.

Galectin-9 recognizes and exhibits antimicrobial activity toward microbes expressing blood group–like antigens

While adaptive immunity recognizes a nearly infinite range of antigenic determinants, immune tolerance renders adaptive immunity vulnerable to microbes decorated in self-like antigens. Recent studies suggest that sugar-binding proteins galectin-4 and galectin-8 bind microbes expressing blood group antigens. However, the binding profile and potential antimicrobial activity of other galectins, particularly galectin-9 (Gal-9), has remained incompletely defined. Here, we demonstrate that while Gal-9 possesses strong binding preference for ABO(H) blood group antigens, each domain exhibits distinct binding patterns, with the C-terminal domain (Gal-9C) exhibiting higher binding to blood group B than the N-terminal domain (Gal-9N). Despite this binding preference, Gal-9 readily killed blood group B–positive Escherichia coli, whereas Gal-9N displayed higher killing activity against this microbe than Gal-9C. Utilization of microarrays populated with blood group O antigens from a diverse array of microbes revealed that Gal-9 can bind various microbial glycans, whereas Gal-9N and Gal-9C displayed distinct and overlapping binding preferences. Flow cytometric examination of intact microbes corroborated the microbial glycan microarray findings, demonstrating that Gal-9, Gal-9N, and Gal-9C also possess the capacity to recognize distinct strains of Providencia alcalifaciens and Klebsiella pneumoniae that express mammalian blood group–like antigens while failing to bind related strains that do not express mammalian-like glycans. In each case of microbial binding, Gal-9, Gal-9N, and Gal-9C induced microbial death. In contrast, while Gal-9, Gal-9N, and Gal-9C engaged red blood cells, each failed to induce hemolysis. These data suggest that Gal-9 recognition of distinct microbial strains may provide antimicrobial activity against molecular mimicry.

Molecular Characterization of Bacteria Associated with Vended Suya Meat in Port Harcourt

The contamination of vended food with microorganisms especially pathogenic microbes is a public health hazard that could result to gastroenteritis. The aim of this study was to identify by molecular techniques bacteria associated with vended suya meat in part of Port Harcourt. Forty (40) ready-to-eat suya meat were randomly bought from 10 vendors across four locations: Rumuokoro, Rukpokwu, Nkpolu, and Choba. Sampling was carried out for a period of 3 months (September to November, 2019). The total heterotrophic bacterial counts of the vended suya meat for Rumuokoro, Rukpokwu, Nkpolu, and Choba were 1.04×106, 3.4×106, 1.49×106 and 2.04×106 CFU/g, respectively. While the total coliform counts of the vended suya meat for Rumuokoro, Rukpokwu, Nkpolu and Choba were 0.36×105, 3.21×105, 2.45×105 and 6.39×105 CFU/g, respectively. The total heterotrophic bacterial counts of vended suya meat bought from vendors in Choba were significantly higher (P≤ .05) than those bought from the Rumuokoro and Rukpokwu vendors. Similarly, the coliform counts of the suya meat bought from Choba vendors were significantly higher (P≤ .05) higher than the coliform counts of vended suya meat bought from vendors in Rumuokoro and Nkpolu. Twenty-eight bacterial isolates: Staphylococcus delphini, Staphylococcus lugdunensis, Bacillus subtilis, Staphylococcus pasteuri, Paenibacillus pectinolytic, Lysinibacillus fusiforms, Bacillus aerius, Serratia nematodephila, Providencia alcalifaciens, Klebsiella singaporensis, Pseudomona aeruginosa, E. coli, Pseudomonas fluorescens and Proteus myxofaciens were identified from the vended suya meat. The molecular characterization of 16S rRNA of the isolates showed 99-100% similarity to other species in the NCBI data base. The evolutionary distances computed were in agreement with the phylogenetic placement of the 16S rRNA of the isolates Providencia and the Bacillus spp respectively and revealed a close relatedness to Providencia stuartii and Bacillus flexus respectively. The 16S rRNA of Bacillus, Pseudomonas, and Lysinibacillus spp revealed a close relatedness to Bacillus flexus, Pseudomonas aeruginosa, and Lysinibacillus fusiformis. The frequency of occurrence of bacterial isolates across the locations was: Pseudomonas aeruginosa (7.14), Bacillus flexus (7.14), Bacillus spp (14.29), Staphylococcus sp (14.29), Staphylococcus lugdunensis (10.71), Proteus sp (10.71), Lynsibacillus macroides (3.57), E. coli (10.71), Serratia spp (10.71), Klebsiella spp (7.14) and Providencia alcalifaciens (3.57). These bacterial genera could pose serious health challenges especially if they are consumed in quantities required to cause infections as many have been linked to causing gastroenteritis and other forms of infections. Proper hygiene compliance during preparation and packaging is recommended to eliminate or reduce microbial populations and types.

Isolation and Characterization of Different Bacterial Strains from Soils, With the Potential of Degrading Polychlorinated Biphenyls (PCBs) Under Aerobic Conditions

Polychlorinated biphenyl (PCBs) are persistent organic compounds used between 1930 and 1970 as thermal insulators in electrical equipment such as transformers, electrical condensers, heat exchangers, and hydraulic systems. Currently, this xenobiotic is a potential source of pollution to soil and water resources. This research aimed to isolate and characterize bacteria strains possessing a high potential to degrade PCBs contained in used dielectric oils. The bacteria strains were isolated from a soil sample taken from “Alao” Hydroelectric Power Center. Four culture media were used for the soil sample culturing: Agar for Standard Methods Medium (STD), Phosphorus, Ammonium, and Salts Medium (PAS), Minimum Medium (Mm), and Minimum Medium (Mm) supplemented with 25 µL of oil contaminated with 43,6 ppm of PCBs. The isolation, screening, and stabilization process were carried out using three culture media: STD, PAS, and Mm, each supplemented with 25 µL of oil contaminated with 43,6 ppm of PCBs. Nineteen bacterial colonies were isolated and exposed to a higher concentration of PCBs. Eighteen colonies could tolerate 250 µL of oil contaminated with 500 ppm of PCBs. Once stabilized and purified these bacteria strains, were characterized according to their microscopic and macroscopic characteristics and tolerance to heavy metals. Through biochemical tests, 18 strains were identified belonging to Providencia alcalifaciens, Proteus mirabillis, Providencia rettgeri, Providencia stuartii, Pseudomona alcaligenes, Morganella morganii, and Shigella disenteriae

High burden of multidrug resistant bacteria detected in Little Akaki River

Spread of antimicrobial-resistant bacteria between humans and animals occurs when the environment is contaminated with animal and human wastes. A total of 30 samples were collected from the Akaki river to identify antimicrobial-resistant bacteria. Bacterial enumeration and characterization was done by spreading serially diluted water samples on MacConkey agar. Sixty four bacterial isolates were identified and susceptibility tested using VITEK 2. The most frequently identified bacteria were Providencia alcalifaciens 10 (15.6%), Kluyvera cryocrescens 9 (14.1%) and Citrobacter freundii 7(10.9%), respectively. Multiple drug resistant bacteria were identified, constituting 17 (28%) of the 64 identified isolates. Multiple antimicrobial resistance (MAR) index of the six sites laid in the range 0.13–0.27, being the highest score located downstream of all the sampling sites. Species MAR index varied from 0.12 to 0.40. Out of 64 isolates, 54 (84.4%) of them were resistant to Ampicillin. On the contrary, most of the isolates were sensitive to Amikacin and meropenem. In conclusion, our findings indicated E.coli count was above the WHO permissible levels. The predominant isolates were P. alcalifaciens, and C. freundii. The MAR index of major isolates was greater than two, implying the study sites were exposed to high-risk sources of human or animal contamination.

An Official Outbreak Investigation of Acute Haemorrhagic Diarrhoea in Dogs in Norway Points to Providencia alcalifaciens as a Likely Cause.

Simple SummaryAn official outbreak investigation to reveal the cause and possible common exposures of dogs suffering acute haemorrhagic diarrhoea (AHD) was performed by the veterinary authorities in Norway in 2019. The outbreak had been reported by private veterinarians who were consulting a greater number of dogs than usual with severe AHD. Epidemiological and diagnostic investigations pointed to the bacteria Providencia alcalifaciens as a possible cause of the outbreak. Whole genome sequencing of bacterial strains from 51 dogs showed that they were almost identical, which implies that the dogs had been exposed to a common source of infection. However, epidemiological investigations did not reveal a common source. Further studies are needed to investigate the disease-causing properties of P. alcalifaciens in dogs.An outbreak investigation was initiated in September 2019, following a notification to the Norwegian Food Safety Authority (NFSA) of an unusually high number of dogs with acute haemorrhagic diarrhoea (AHD) in Oslo. Diagnostic testing by reporting veterinarians had not detected a cause. The official investigation sought to identify a possible common cause, the extent of the outbreak and prevent spread. Epidemiological data were collected through a survey to veterinarians and interviews with dog owners. Diagnostic investigations included necropsies and microbiological, parasitological and toxicological analysis of faecal samples and food. In total, 511 dogs with acute haemorrhagic diarrhoea were registered between 1 August and 1 October. Results indicated a common point source for affected dogs, but were inconclusive with regard to common exposures. A notable finding was that 134 of 325 faecal samples (41%) cultured positive for Providencia alcalifaciens. Whole genome sequencing (WGS) of 75 P. alcalifaciens isolates from 73 dogs revealed that strains from 51 dogs belonged to the same WGS clone. Findings point to P. alcalifaciens as implicated in the outbreak, but investigations are needed to reveal the pathogenic potential of P. alcalifaciens in dogs and its epidemiology.

Molecular Characterization of Gut Bacterial Flora of Honeybee (Apis Mellifera Adansonii) From Some Selected Apiaries in Ogun State, Nigeria

Research related to physiology and pathology of honey bees in particular Apis mellifera adansonii has attracted a lot of attention. The present study is aimed to determine honeybee (Apis mellifera adansonii) gut microbiome from Apiary in Olabisi Onabanjo University and Osoba Avenue Odo-Epo, Odogbolu Local Government. Twenty (20) honeybees workers (A. mellifera) were collected into a small vile containing sugar powder from the apiary located in OOU and Osoba Avenue at Odo-Epo during rainy season in July and transported to Zoology and Environmental Biology laboratory in OOU and kept in ice-cubes (-50C) till daybreak. Standard microbiological analysis for isolation of bacteria was used, adopting Clinical Laboratory Standard Institute procedures. The phylogenetic analyses based on the 16S rDNA gene were further used to characterize the organism in order to establish relationships among them. The results showed microbiota of the studied samples includes; Cedeca davisae, Cronobacter dublinensis, Enterobacter aerogenes, Kluyvera cryocrescens, Klebsiella oxytoca, Providencia vermicola, Salmonella enteric, Providencia alcalifaciens, Serratia nematodiphila, Pseudomonas plecoqlossicida, Klebsiella michiganensis, Serratia marcenscens, Pseudomonas aeruginosa, Escherichia coli, Aeromonas hydrophila and Enterobacter asburiae. Klebsiella spp. was more abundant and prominent in the digestive guts of honeybee workers both in OOU and Osoba Avenue, Odo Epo. The result of the percentage identity and closest accession of the isolates revealed that, Enterobacter aerogenes had the closest accession number and with highest percentage identity of (99%). The findings from this study showed that microbiota component communities of A. mellifera adansonii in OOU were composed of more Gram-negative bacteria than Gram-positive bacteria in Odo Epo.

Changes in the fecal microbiota in dogs with acute hemorrhagic diarrhea during an outbreak in Norway.

BackgroundA severe form of acute hemorrhagic diarrhea syndrome (AHDS) occurred in dogs in the Oslo region of Norway during autumn 2019.ObjectivesTo characterize the fecal microbiota of dogs with AHDS during the outbreak and compare it to that of healthy dogs from the same period and before the outbreak.AnimalsDogs with AHDS (n = 50), dogs with nonhemorrhagic diarrhea (n = 3), and healthy dogs (n = 11) were sampled during the outbreak. In addition, 78 healthy dogs from the same region were sampled before the outbreak between 2017 and 2018.MethodsRetrospective case‐control study. The fecal microbiotas were characterized using 16S rRNA gene amplicon sequencing.ResultsDogs with AHDS had significantly different microbiota composition (R 2 = .07, P < .001) and decreased intestinal diversity relative to healthy dogs from the outbreak period (median, 2.7; range, 0.9‐3.5 vs median, 3.2; range, 2.6‐4.0; P < .001). The microbiota in dogs with AHDS was characterized by a decrease of Firmicutes and an outgrowth of Proteobacteria, with increased numbers of Clostridium perfringens and Providencia spp. Among the Providencia spp., 1 showed 100% sequence identity with a Providencia alcalifaciens strain that was cultivated and isolated from the same outbreak. No Providencia spp. was found in healthy dogs sampled before the outbreak.Conclusions and Clinical ImportanceDogs with AHDS had marked changes in fecal microbiota including increased numbers of Providencia spp. and C. perfringens, which may have contributed to the severity of this illness.

Association of the microbiota dysbiosis in the hepatopancreas of farmed crayfish (Procambarus clarkii) with disease outbreaks

Symbiotic microorganisms in the hepatopancreas have been found in various crustaceans, and their vital role in maintaining host health has drawn increasing attention. However, the information about the symbiotic microbiota in the hepatopancreas of freshwater crayfish (Procambarus clarkii) is scarce. In this study, the high-throughput sequencing technology was used to characterize the bacterial communities in the hepatopancreas of P. clarkii at different health statuses, including healthy, anorexic, moribund, and whitish muscle statuses. Results showed that the hepatopancreatic microbiota of healthy crayfish was dominated by the phyla Firmicutes, Proteobacteria, and Bacteroidota and Clostridium sensu stricto 11, Proteus, C. sensu stricto 1, Paraclostridium, and Weissella. Distinct differences were found in the structure, composition, and predicted function of the hepatopancreatic microbiota between the healthy and the sick crayfish. The LEfSe analysis revealed that the symbiotic bacterial species, including Proteus penneri, C. sensu stricto 11 and Lactococcus garvieae, and the potential probiotics, such as Weissella cibaria and Lactobacillus murinus, in the hepatopancreas of healthy crayfish were significantly enriched than those of sick crayfish, whereas the opportunistic pathogens, including Citrobacter freundii, Plesiomonas shigelloides, C. sensu stricto 7, and Terrisporobacter, in the hepatopancreas of sick crayfish were significantly enriched than those of healthy crayfish. In addition, compared with that of healthy crayfish, the hepatopancreas of moribund crayfish had significantly enriched bacterial genera, such as Dubosiella, Candidatus, Bacilloplasma, and Phreatobacter, whereas the hepatopancreas of crayfish with whitish muscle disease was observed with a significant enrichment of some opportunistic pathogens, including Morganella morganii, Providencia alcalifaciens, Vagococcus fluvialis, Clostridium lundense, and Bacteroides. This apparent dysbiosis, which showed the increase in opportunistic pathogens and the decrease in symbiotic microorganisms (including potential probiotics), might represent a specific microbial signature of disease occurrence. Furthermore, the PICRUSt2 analysis revealed that the microbial functions involved in carbohydrate metabolism and xenobiotic biodegradation in the hepatopancreas of sick crayfish were significantly enriched than those of healthy crayfish, whereas the microbial functions involved in amino acid metabolism, lipid biosynthesis, biosynthesis of other secondary metabolites, energy metabolism, nucleotide metabolism, transport and catabolism, and immune system in the hepatopancreas of healthy crayfish were significantly enriched compared with those of sick crayfish. This study revealed the hepatopancreatic microbiota dysbiosis associated with disease outbreaks, providing potential indicators for assessing the health status of crayfish and new clues for exploring the potential role of the hepatopancreatic microorganisms in affecting host health and disease.

Comparative Genomic Analysis Reveals Genetic Mechanisms of the Variety of Pathogenicity, Antibiotic Resistance, and Environmental Adaptation of Providencia Genus.

The bacterial genus Providencia is Gram-negative opportunistic pathogens, which have been isolated from a variety of environments and organisms, ranging from humans to animals. Providencia alcalifaciens, Providencia rettgeri, and Providencia stuartii are the most common clinical isolates, however, these three species differ in their pathogenicity, antibiotic resistance and environmental adaptation. Genomes of 91 isolates of the genus Providencia were investigated to clarify their genetic diversity, focusing on virulence factors, antibiotic resistance genes, and environmental adaptation genes. Our study revealed an open pan-genome for the genus Providencia containing 14,720 gene families. Species of the genus Providencia exhibited different functional constraints, with the core genes, accessory genes, and unique genes. A maximum-likelihood phylogeny reconstructed with concatenated single-copy core genes classified all Providencia isolates into 11 distant groups. Comprehensive and systematic comparative genomic analyses revealed that specific distributions of virulence genes, which were highly homologous to virulence genes of the genus Proteus, contributed to diversity in pathogenicity of Providencia alcalifaciens, Providencia rettgeri, and Providencia stuartii. Furthermore, multidrug resistance (MDR) phenotypes of isolates of Providencia rettgeri and Providencia stuartii were predominantly due to resistance genes from class 1 and 2 integrons. In addition, Providencia rettgeri and Providencia stuartii harbored more genes related to material transport and energy metabolism, which conferred a stronger ability to adapt to diverse environments. Overall, our study provided valuable insights into the genetic diversity and functional features of the genus Providencia, and revealed genetic mechanisms underlying diversity in pathogenicity, antibiotic resistance and environmental adaptation of members of this genus.

Influence of inoculated gut bacteria on the development of Bactrocera dorsalis and on its susceptibility to the entomopathogenic fungus, Metarhizium anisopliae

BackgroundSymbiotic interactions between insects and bacteria have been associated with a vast variety of physiological, ecological and evolutionary consequences for the host. A wide range of bacterial communities have been found in association with the oriental fruit fly, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae), an important pest of cultivated fruit in most regions of the world. We evaluated the diversity of gut bacteria in B. dorsalis specimens from several populations in Kenya and investigated the roles of individual bacterial isolates in the development of axenic (germ-free) B. dorsalis fly lines and their responses to the entomopathogenic fungus, Metarhizium anisopliae.ResultsWe sequenced 16S rRNA to evaluate microbiomes and coupled this with bacterial culturing. Bacterial isolates were mono-associated with axenic B. dorsalis embryos. The shortest embryonic development period was recorded in flies with an intact gut microbiome while the longest period was recorded in axenic fly lines. Similarly, larval development was shortest in flies with an intact gut microbiome, in addition to flies inoculated with Providencia alcalifaciens. Adult B. dorsalis flies emerging from embryos that had been mono-associated with a strain of Lactococcus lactis had decreased survival when challenged with a standard dosage of M. anisopliae ICIPE69 conidia. However, there were no differences in survival between the germ-free lines and flies with an intact microbiome.ConclusionsThese findings will contribute to the selection of probiotics used in artificial diets for B. dorsalis rearing and the development of improved integrated pest management strategies based on entomopathogenic fungi.

Determining the prevalence, identity and possible origin of bacterial pathogens in soil.

Soil biomes are vast, exceptionally diverse and crucial to the health of ecosystems and societies. Soils also contain an appreciable, but understudied, diversity of opportunistic human pathogens. With climate change and other forms of environmental degradation potentially increasing exposure risks to soilborne pathogens, it is necessary to gain a better understanding of their ecological drivers. Here we use the Galleria mellonella insect virulence model to selectively isolate pathogenic bacteria from soils in Cornwall (UK). We find a high prevalence of pathogenic soil bacteria with two genera, Providencia and Serratia, being especially common. Providencia alcalifaciens, P. rustigianii, Serratia liquefaciens and S. plymuthica strains were studied in more detail using phenotypic virulence and antibiotic resistance assays and whole-genome sequencing. Both genera displayed low levels of antibiotic resistance and antibiotic resistance gene carriage. However, Serratia isolates were found to carry the recently characterized metallo-β-lactamase blaSPR-1 that, although not conferring high levels of resistance in these strains, poses a potential risk of horizontal transfer to other pathogens where it could be fully functional. The Galleria assay can be a useful approach to uncover the distribution and identity of pathogenic bacteria in the environment, as well as uncover resistance genes with an environmental origin.

Nickel Oxide Nanoparticles Application for Enhancing Biogas Production Using Certain Wastewater Bacteria and Aquatic Macrophytes Biomass

Biogas is one of the practical vitality sources of energy that can be delivered from numerous sorts of biomass. Biogas production from fresh digestate bacterial crude culture; Providencia alcalifaciens and Alcaligenes aquatilis isolated from municipal wastewater treatment plants located in Port Said, Egypt and identified with 16S rRNA; amended with aquatic macrophytes; Pistia stratiotes and Potamogeton pectinatus, collected from some irrigation canals in Port Said, Egypt as a substrate in the presence of different concentrations of chemically prepared nickel oxide nanoparticles (NiONPs) subjected to anaerobic conditions were successfully investigated. The produced biogas was collected by the water displacement method and subsequently measured using GC/MS analysis. As found in this present work, the optimum NiONPs concentration was 2 ppm for biogas production, where the highest productivity was 42.90 mL and 31.33 mL/3.4 g of P. stratiotes and P. pectinatus with enhancing % 87.91% and 82.38%, respectively compared with the control (without NiONPs) at 30 days of incubation period. Biogas production was slightly greater using P. stratiotes than P. pectinatus particularly with the addition of NiONPs. The present examination may have a noteworthy task in the generation of reasonable biogas amounts utilizing monetary wellsprings of reusing the horrible water system trenches macrophytes and control of environmental defilement issues that may have come about in light of these unfortunate macrophytes.

CexE Is a Coat Protein and Virulence Factor of Diarrheagenic Pathogens.

CexE is a 12 kDa protein that was originally reported to be present in just three strains of enterotoxigenic Escherichia coli (ETEC); a frequent cause of diarrheal illnesses worldwide. However, an examination of sequenced genomes has revealed that CexE is actually present in a majority of ETEC strains. In addition, homologs of CexE are present in enteroaggregative E. coli (EAEC), Yersinia enterocolitica, Providencia alcalifaciens, and Citrobacter rodentium. Although it has been hypothesized that CexE and its homologs are virulence factors, this has yet to be tested. Thus the primary aim of this study was to determine if these proteins contribute to pathogenicity. Our secondary aim was determine if they are secreted coat proteins. Here we report that all neonatal mice infected with a wild-type strain of C. rodentium perished. In contrast a cexE mutant was significantly attenuated with 45% neonate survival. In adult mice the wild-type strain reached significantly higher loads in the large intestines and were shed in greater numbers than cexE mutants. Secretion of the CexE homolog in EAEC is dependent upon an atypical Type I secretion system that accepts its client from the periplasm rather than the cytoplasm. Insertion mutants of cexC, the putative ATPase of the CexE secretion system, were attenuated in our murine model. In vitro we found that CexC is required for the secretion of CexE to the outer membranes of both ETEC and C. rodentium. Secretion is not constitutive because CexE accumulates in the periplasm when the two pathogens are cultured under noninducing conditions. Although secretion conditions differ between ETEC and C. rodentium, secreted CexE remains predominantly associated with the outer membranes of both species. In aggregate these findings demonstrate that CexE is a secreted coat protein and virulence factor that promotes colonization of host intestinal tissues by enteric pathogens.

Screening and Characterization of Antibiotic Producing Organisms from Waste Dump Soil Sample

This study was designed to screen and characterize antibiotic producing bacteria and fungi from soil samples from a waste dump site in Enugu. The molecular characterization of the isolates was performed by using Zymo Research Quick-DNA fungi/bacteria miniprep kit. The bacteria isolates were Providencia stuartii, Providencia alcalifaciens. Bacillus sp., Streptococcus sp., Micrococcus sp., and Staphylococcus sp while Aspergillus fumigatus and Aspergillus niger were fungal isolates. The efficacy of the Crude Extract (CE) of the antibiotics produced was tested against species of Staphylococcus aureus and Escherichia coli. The CE of antibiotics when compared with a conventional antibiotic (Ciprofloxacin) showed lower antimicrobial activity. The minimum inhibitory concentration of the crude extracts ranged between 12.5% and 25% concentrations while the minimum bactericidal concentration ranged from 25% and 50% concentrations. The findings suggest that these organisms have the potential to produce antibiotics and could be harnessed by pharmaceutical industries for the production of newer antibiotics.

Screening for Bio-flocculant Producing Bacterial Strains from Asa River in Ilorin Kwara State

Abstract Synthetic flocculants in water treatment have been reported to be detrimental to both human health and the environment. Thus, there is a constant search for bio-flocculants that is safe and addresses the effects of synthetic polymers. This study was aimed at isolating bacteria with bio-flocculating potential, their molecular identification and phylogenetic relatedness, and the optimization of their flocculating abilities from an open water (Asa River) in Ilorin Kwara State. The effect of carbon source (glucose, lactose and starch), cations (FeSO4, KCl and CaCl2) and pH (2 to 12) was evaluated on the bio-flocculating activities of the isolates using kaolin clay. The seven (7) novel (new strains) isolates with their respective accession number from National Center for Biotechnology Information (NCBI) after molecular confirmation are Pseudomonas otitidis MTK01 (MK263227), Aeromonas caviea MTK02 (MK263228), Providencia alcalifaciens MTK03 (MK263229), Providencia sp. MTK05 (MK263230), Alcaligenes sp. MTK06 (MK263231), Klebsiella pneumoniae MTK07 (MK263232) and Klebsiella sp. MTK08 (MK263233) while Raoultella ornithinolytica MTK04 was also identified. Phylogenetic tree of relatedness showed close kin of the isolates to established bacteria sequence deposited at the NCBI GenBank. Although the flocculating rate of each isolates varied with different parameters that was used in the study, glucose was the most supportive followed by lactose and starch, CaCl2 was most supportive cation followed by KCl and FeSO4 while pH 12, 6, 8, 10, 2 and 4 were the order of decreasing flocculating rate of the medium. This study has reported the presence of eight (8) bio-flocculating bacteria (out of which 7 are new strains of bacteria) in an open water which has been further optimized for effective flocculating rate and thus provides an ecofriendly and harmless flocculants source that can be employed in water treatment procedures.