Serratia Fonticola Research Articles

Antibiotic concentrations induce morphological changes and increase biofilm formation in multi-antibiotic and heavy metal resistant Kluyvera cryocrescens and Serratia fonticola

Water pollution is the biggest challenge that has rendered existing water resources unusable due to contamination with antibiotics and heavy metals. Antibiotics are often used to treat bacterial diseases. Heavy metals, on the other hand, are micro-pollutants that pose a threat to aquatic systems, especially when they accumulate in nature. Increasing pollution and the uncontrolled use of antibiotics have exposed bacteria to non-lethal concentrations (sub-MIC), potentially leading to resistance. In this study, Kluyvera cryocrescens and Serratia fonticola were isolated from a freshwater source and characterised. The resistance profiles of the isolates to 16 antibiotics and 8 heavy metals were determined, revealing that they are multidrug-resistant. The effects of sub-MICs (MIC/2 and MIC/4) of antibiotics on biofilm formation, siderophore production, and cell morphology of bacteria were analysed. It was found that at some sub-MIC values of kanamycin, tetracycline, meropenem, erythromycin, and clarithromycin, biofilm formation by K. cryocrescens increased. An increase in biofilm production was also observed in S. fonticola at sub-MIC values of imipenem, meropenem, ceftazidime, ciprofloxacin, and clarithromycin. Moreover, significant morphological changes were observed in both isolates following treatment with meropenem, ciprofloxacin, and ceftazidime. After treatment with meropenem, the typical rod-shaped (bacillary) morphology of the isolates shifted to a round (coccoid) form. In contrast, the bacteria developed into long filaments after treatment with ciprofloxacin and ceftazidime. These changes in the bacteria may favour the development of resistance and pose challenges for the prevention and treatment of diseases. Therefore, it is crucial to understand how sub-MIC levels of antimicrobial agents alter the virulence properties of bacteria.

Enhancing tomato growth and soil fertility under salinity stress using halotolerant plant growth-promoting rhizobacteria

Soil salinization is a critical issue that not only hampers the efficiency and sustainability of global agricultural production but also poses significant challenges to the achievement of sustainable development goals across environmental, economic, and social dimensions. Halotolerant plant growth-promoting rhizobacteria (HPGPR) have the potential to mitigate abiotic stress, foster plant growth, and bolster the stress resistance capabilities of crops. This study conducted the isolation, identification, and characterization of HPGPR originating from a saline-alkali orchard area in northwest China. The efficacy of the isolated bacterial strains was evaluated through potted plant experiments, assessing the growth of tomato plants under in vitro conditions and under varying salinity stress. Ultimately, the study investigated the influence of these HPGPR on soil physicochemical properties, enzymatic activities, and the structure and composition of the microbial community. Upon isolating 12 bacterial strains, we conducted an in vitro assessment of their salt tolerance, ultimately singling out three robust isolates, which exhibited exceptional salt tolerance. Detailed 16S rRNA gene sequencing and meticulous taxonomic evaluation systematically assigned these isolates to Priestia endophyticus GSCK1 (accession number: OR569048), Bacillus atrophaeus GSCK2 (accession number: OR569061), and Serratia fonticola GSCK6 (accession number: OR569062), respectively. These strains exhibited notable biochemical and plant growth-promoting traits, including enzymatic activities and the production of indole-3-acetic acid. They significantly enhanced plant growth metrics and soil fertilities, particularly strain GSCK6, which also reshaped the soil microbial community, augmenting beneficial microbe abundance. The HPGPR treatment notably improved soil pH, nutrient availability, enzymatic activities, and reduced soil electrical conductivity, underscoring their potential in agricultural resilience against salinity. The eco-friendly salt stress mitigation strategy of HPGPR not only enhances soil quality and promotes plant growth by regulating the composition and function of microbial communities, but also provides a novel solution for global agricultural production. This approach is conducive to increasing crop yield and quality, reducing the limitations of saline-alkali land on agricultural production, and promoting food security and sustainable agricultural development.

Extended-spectrum ß-lactamase (ESBL)- and Carbapenemase-producing Enterobacterales Isolated from Fresh Herbs and Salads at Retail Level in Switzerland

Fresh produce is usually consumed raw or minimally processed, making it a potential vehicle for the transmission of antimicrobial-resistant (AMR) microorganisms to humans. The objective of the study was to assess the occurrence of extended-spectrum ß-lactamase (ESBL)- and carbapenemase-producing Enterobacterales (ESBL-E and CPE), respectively, in 118 fresh herbs and 101 bagged salads collected at retail level in Switzerland and to characterize the isolates’ phenotypic and genotypic properties using culture-based methods and whole genome sequencing (WGS).Of the fresh herbs, 6/118 contained ESBL-E and 7/118 yielded CPE. Of the salads, 13/101 contained ESBL-E and 1/101 CPE. Antimicrobial susceptibility testing (AST) identified 9/29 isolates as multidrug-resistant (MDR).ESBL-E were Escherichia coli (n = 6), Klebsiella pneumoniae (n = 4) Enterobacter chuandaensis (n = 1), and Kluyvera spp. (n = 1) carrying ß-lactamase (bla) genes belonging to the cefotaximase-München (blaCTX-M)-groups, Proteus spp. (n = 1) containing Hôpital-Universitaire-de-Genève-bla (blahugA), Raoultella ornithinolytica (n = 1) carrying sulfhydryl reagent variable bla (blaSHV), and Serratia fonticola (n = 7) carrying S. fonticula bla (blaFONA) genes. CPE were Enterobacter asburiae (n = 1) E. cloacae (n = 6) and E. vonholyi (n = 1) carrying imipenemase bla (blaIMI) genes.Several K. pneumoniae sequence types (STs) were identified (ST967, ST628, ST219, and ST1823), which have been linked to human disease and nosocomial outbreaks. They carried blaCTX-M-15 on plasmids detected globally in environmental and clinical samples. E. coli (ST10, ST48, ST609, ST2040, ST6215 and ST3580) and enterotoxigenic E. coli (ETEC) ST2040 carrying blaCTX-M-15 were found. E. cloacae (ST820 and ST1516) with blaIMI-1 have been found previously in clinical settings and community outbreaks.The occurrence and consumption of fresh produce containing MDR ESBL-E and CPE pose substantial public health risks and raise significant food safety concerns.

Comparison of antimicrobial resistant Enterobacterales isolates from the dairy production environment in low and high zinc containing regions

Heavy metals occur naturally in the environment, and their concentration varies in soil across different regions. However, the presence of heavy metals may influence the antimicrobial resistance (AMR) in bacterial populations. Therefore, the objective of this study was to investigate and characterise the antimicrobial resistance profiles of Enterobacterales in soil and bovine milk filters from high and low zinc-containing regions in Ireland. In total, 50 soil samples and 29 milk filters were collected from two geographic locations with varying soil zinc concentrations. Samples were cultured for the enumeration and detection of Enterobacterales. Specifically, extended-spectrum beta-lactamase-producing Enterobacterales, carbapenem-resistant Enterobacterales and ciprofloxacin-resistant Enterobacterales were isolated using selective media. Species identification was performed using MALDI-TOF. The phenotypic resistance profiles of selected Enterobacterales were determined by disk diffusion testing, following EUCAST and CLSI criteria; while, the genotypic resistance profiles of the same isolates were determined by whole genome sequencing (WGS). Heavy metal concentrations were also measured for all soil samples. A total of 40 antimicrobial resistant Enterobacterales were identified in soil (n = 31) and milk filters (n = 9). The predominant species detected in the high zinc-containing region was Escherichia coli in both sample types (soil n = 10, milk filters n = 2), while in the low zinc-containing region Serratia fonticola was predominant in soil samples (n = 8) and E. coli in milk filters (n = 4). Ten E. coli isolates identified from soil samples in the high zinc-containing region were multidrug resistant, showing resistance to all the antimicrobials tested, except for carbapenems. The WGS findings confirmed the phenotypic resistance results. Moreover, zinc resistance-associated genes and genes encoding for efflux pumps were identified. The current study revealed distinct phenotypic resistance profiles of Enterobacterales in low and high zinc-containing regions, and highlighted the benefit of utilising milk filters for AMR surveillance in dairy production.

Characterization of antimicrobial resistant Enterobacterales isolated from spinach and soil following zinc amendment

Antimicrobial resistant bacteria can occur in the primary food production environment. The emergence and dissemination of antimicrobial resistance (AMR) in the environment can be influenced by several factors, including the presence of heavy metals. The aim of this study was to examine the presence and characteristics of antimicrobial resistant Enterobacterales in soils and spinach grown in soils with and without zinc amendment. A total of 160 samples (92 soil and 68 spinach) were collected from two locations, in which some plots had been amended with zinc. Samples were cultured on selective agars for detection of extended-spectrum beta-lactamase-producing Enterobacterales (ESBL), carbapenem-resistant Enterobacterales and ciprofloxacin-resistant Enterobacterales. Samples were also cultured for enumeration of total Enterobacterales. Isolates were identified by MALDI-TOF. Antimicrobial susceptibility testing was carried out in accordance with EUCAST and CLSI criteria. The whole genome sequence (WGS) of selected isolates was determined. Inductively coupled plasma atomic emission spectrometry was also performed on soil samples in order to measure the concentration of zinc. In total 20 antimicrobial resistant Enterobacterales were isolated from the soil (n = 8) and spinach samples (n = 12). In both sample types, Serratia fonticola (n = 16) was the dominant species, followed by Escherichia coli (n = 1), Citrobacter freundii (n = 1) and Morganella morganii (n = 1) detected in spinach samples, and Enterobacter cloacae (n = 1) detected in a soil sample. The WGS identified genes conferring resistance to different antimicrobials in agreement with the phenotypic results; 14 S. fonticola isolates were confirmed as ESBL producers and harboured the blaFONA gene. Genes that encoded for zinc resistance and multidrug efflux pumps, transporters that can target both antimicrobials and heavy metals, were also identified. Overall, the findings of this study suggest the presence of zinc did not influence the AMR Enterobacterales in soil or spinach samples.

Biosynthesis of Iron Oxide Nanoparticles and Combination with Glyphosate Herbicide and Effect Against Cogon Grass Control

For enhancing herbicide efficacy a total of 42 soil samples, five species of bacteria that grew the most were selected for regarding other isolates: (Enterobacter cloacae, Serratia fonticola, Aeromonas hydrophila, Pantoea spp., Pseudomonas mendocina). Extracellular extracts of these bacteria were made and then used to manufacture iron oxide nanoparticles biologically. It was observed after measuring the average diameter by AFM (Atomic Force Microscopy) for each NPs prepared were 65.14, 29.21, 63.87, 57.89 and 36.59 nm, respectively. The synthesis conditions were a pH of 7 and 50°C. Based on the AFM values, Serratia fonticola was selected as having the smallest average diameter of 29.21 nm to complete the rest of the techniques (UV-VIS, AFM, FTIR and FE-SEM). The wavelength of Fe2O3NPs by using UV-VIS is 359 nm, Image FE-SEM displays spherical Fe2O3NPs in Nano-cluster form and the average volume is 29.21 nm by AFM. Utilizing the Randomized Complete Block Design methodology with 6 treatments, two factors: Glyphosate herbicide at the recommended dose and three concentrations of the prepared (Fe2O3NPs) from Serratia fonticola were 0.5, 1 and 1.5 g/ml. Halfa plant was controlled using these treatments, and after 30 days, the efficiency of the treatments was measured on a set of characteristics. It was found the lowest number of plants and the highest percentage of control was in the treatment 5 glyphosate and Fe2O3 NPs con. (1.5g/ml) was 1.26, 5.33 and 90%, respectively. The study effectively shows the potential of employing Fe2O3NPs generated from S. fonticola to improve the efficacy of Glyphosate for Cogon management.

Characteristics of Genomes of Opportunistic Bacteria in Zoogenic Ecosystems of the Russian Western Arctic Islands

Introduction: Arctic ecosystems are the most important object of microbiological surveillance that helps monitor a rapidly changing natural environment affected by climate change and industrial development of the northern territories. Objective: To assess the epidemic potential of opportunistic bacteria associated with animals from the Arctic islands of the Barents and Kara Seas. Materials and methods: We have studied four strains of gram-negative opportunistic bacteria (Serratia fonticola, Aeromonas salmonicida, Yersinia kristensenii, and Yersinia rochesterensis) isolated in 2021–2022 during microbiological testing of 46 samples of zoogenic biologic materials from the natural ecosystems of Vaigach Island, Novaya Zemlya and Franz Josef Land archipelagos. Genomes of the isolated microbial strains were sequenced using the Illumina MiSeq sequencing system and, after de novo assembly using the SPAdes 13.0 genomic assembler, annotated with RAST (Rapid Annotation using Subsystem Technology). Results: The strains were shown to possess a number of virulence factors and antibiotic resistance genes, which allows them to be considered as potential pathogens. Classes A, B, and C beta-lactamases were found in the genomes of all the bacteria under study, and operons for siderophores and type IV secretion systems were typical of virulence factors. In addition, the pertussis-like YtxA enterotoxin was identified in Yersinia genomes. Conclusions: Our findings indicate the possibility of emergence of natural foci of notifiable infections on the islands in the high-latitude Arctic promising in terms of tourism development and economic growth.

Impact of operational conditions on drinking water biofilm dynamics and coliform invasion potential.

The revelation that even low concentrations of coliforms can infiltrate into mature drinking water biofilms highlights a potential public health concern. Nowadays, the measurement of coliform bacteria is used as an indicator for fecal contamination and to control the effectiveness of disinfection processes and the cleanliness and integrity of distribution systems. In Flanders (Belgium), 533 out of 18,840 measurements exceeded the established norm for the coliform indicator parameter in 2021; however, the source of microbial contamination is mostly unknown. Here, we showed that mature biofilms, are susceptible to invasion of Serratia fonticola. These findings emphasize the importance of understanding and managing biofilms in drinking water distribution systems, not only for their potential to influence water quality, but also for their role in harboring and potentially disseminating pathogens. Further research into biofilm detachment, long-term responses to operational changes, and pathogen persistence within biofilms is crucial to inform strategies for safeguarding drinking water quality.

Studying some of the optimal conditionals for the best bacterial isolates from soil and its effects on their growth rates

The current study aimed to use some bacterial isolates from the local soil of Baghdad city by study the effects of temperature, pH and incubation period on the growth rates of isolated bacteria and choose the optimal conditions for their diversity and for understanding bacterial growth and their requirements for survival and proliferation. This information can be applied to obtain their high growth rate for use in various fields such as agriculture, medicine and environmental sciences in the future. And it used to assess the degree of variation in across bacteria species in pH, temperature and incubation period. A number of local bacterial isolates as Enterobacter cloacae, Aeromonas hydrophila, Pantoea spp., Pseudomonas mendocina and Serratia fonticola identified in this study. The growth rate of the bacterial isolates was tested by growing each bacterial isolate in a different range of pH (4, 7 and 10) and under the influence of several temperatures (52, 35 and 50) and within different incubation periods (24, 48 and 72) hours The optimal conditions were determined by measuring the optical density (absorbance) at a wavelength of 600 nm, which represents bacterial growth. The optimal conditions for E. cloacae were determined to be a pH of 10 and a temperature of 35°C after a 72hour incubation period. Similarly, A. hydrophila exhibited optimal growth at a pH of 7 and a temperature of 35°C after a 72hour incubation period. Pantoea spp. demonstrated optimal growth at a pH of 7 and a temperature of 25°C after a 24hour incubation period. P. mendocina displayed optimal growth at a pH of 7 and a temperature of 25°C after a 72hour incubation period. Lastly, S. fonticola exhibited optimal growth at a pH of 7 and a temperature of 50°C after a 72hour incubation period.

Ecological features of trace elements tolerant microbes isolated from sewage sludge of urban wastewater treatment plant

Worldwide wastewater treatment plants generate enormous amounts of sewage sludge, and their further disposal depends on the treatment technologies applied and spontaneously occurring microbiological processes. From different ages urban sewage sludge, 12 strains of bacteria with simultaneous tolerance to two or more trace elements: Co, Ni, Cu, Zn, Cd and Pb at concentration of 3-5 mmol were isolated and identified by PCR of target genes and Sanger sequencing methods. The isloated metal(loids) tolerant strains belong to the species, i.e., Serratia fonticola, Rhodococcus qingshengii, Pseudomonas fragi, Pseudomonas extremaustralis, Pseudomonas cedrina, Stenotrophomonas maltophilia, Serratia liquefaciens and Citrobacter freundii. The ecological features of the isolated strains were studied. The optimal growth temperatures for most strains was 15–30°C at pH range of 5–9, although some strains grew at 7°C (Pseudomonas fragi SS0-4, Serratia fonticola SS0-9 and Serratia fonticola SS12-11). Satisfactory growth of two strains (Serratia fonticola SS0-1and Citrobacter freundii SS60-12) was noted in an acidic medium at pH 4. Most of the strains grew in the NaCl concentration range of 1–5%. The isolated bacteria resistant to high concentrations of trace elements can be used for the effective mineralization of sewage sludge and for the decontamination of wastewater.

Carbapenemase and extended-spectrum beta-lactamase-producing bacteria in waters originating from a single landfill in Slovenia.

Groundwater, rainwater, and leachate associated with a single landfill were analysed to detect extended-spectrum beta-lactamase (ESBL)-producing and carbapenemase (CP)-producing bacteria. After cultivation on three commercial selective-differential media, 240 bacterial isolates were obtained and identified by matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS). Isolates from clinically relevant species were further genotyped by enterobacterial repetitive intergenic consensus polymerase chain reaction, and tested for antibiotic susceptibility and presence of CPs and ESBL enzymes. Two ESBL-producing isolates and two isolates producing CPs were detected in rainwater, groundwater, and leachate: Klebsiella oxytoca complex with the gene for the ESBL enzyme CTX-M-1 and the gene for the CP OXA-48, Serratia fonticola with the gene for the ESBL enzyme FONA-2, and Pseudomonas aeruginosa with the gene coding Verona integron-encoded Metallo-beta-lactamases (VIM) metallo-beta-lactamase. Our study indicates that bacteria with ESBL and CP genes can be present in landfill-associated waters.

In vitro synthesis and biochemical characterization of acyl-homoserine lactone synthase and its deletion mutant.

Quorum sensing can induce density-dependent gene expressions that cause various problems. For quorum-sensing inhibition, fundamental solutions such as gene manipulation are required, and acyl-homoserine lactone synthase (AHL synthase), which synthesizes the universal quorum-sensing signal of gram-negative bacteria, can be used as a target. In this study, researchers synthesized His-tagged AHL synthase and its deletion mutant that lacks the active site and compared their biochemical characteristics. His-YpeI, the 6x His-tagged AHL synthase of Serratia fonticola, and His-ΔYpeI, its deletion mutant, were designed, and their property conservation were examined using in silico projection tools. For in vitro synthesis of enzymes, the His-YpeI CFPS template was synthesized by in vitro gene synthesis, and the His-ΔYpeI CFPS template was obtained by deletion PCR. CFPS was performed and the products were purified with the 6x His-tag. The enzymes' properties were compared using an enzymatic assay. The bioinformatic analysis confirmed the conservation of biochemical properties between 6x His-tagged and untagged enzymes, including helix-turn-helix interactions, hydropathy profiles, and tertiary structure between His-YpeI and YpeI and between His-ΔYpeI and ΔYpeI. His-YpeI and His-ΔYpeI synthesized by CFPS were found to have the expected molecular weights and demonstrated distinct differences in enzyme activity. The analyzed enzymatic constants supported a significant decrease in substrate affinity and reaction rate as a result of YpeI's enzyme active site deletion. This result showed that CFPS could be used for in vitro protein synthesis, and quorum sensing could be inhibited at the enzymatic level due to the enzyme active site's deletion mutation.

Potential Pathogenicity of Escherichia coli Isolated from the Stools of Healthy Children Suffering from Diarrhea Admitted to Hospitals in Southern Benin

Aims: Infant mortality linked to infectious diseases remains a major public health concern, especially in the context of antibiotic resistance. This work aimed to study the potential pathogenicity of Escherichia coli strains isolated from the stools of children suffering from diarrhea and healthy from 0 to 15 years old admitted to hospitals in southern Benin.
 Methodology: The isolation and characterization of the strains were carried out according to standard microbiology methods. Molecular characterization was carried out by searching for eight virulence factors (Stx1, Stx2, eaeA, bfpB, LT, InVE, IpaH, AggR) and three antibiotic resistance genes (blaTEM, blaSHV, blaCTX -M) by the PCR technique.
 Results: This study shows that 47.06% of the children sampled suffered from diarrhea, against 52.94% who were healthy. A total of 9 species of Enterobacteriaceae were identified in different proportions: Escherichia coli (66.66%), Klebsiella ornithinolytica (13.72%), Klebsiella pneumonia (5.88%), Enterobacter aerogenes (3.92%), Escherichia vulneris (1.96%), Salmonella arizonae (1.96%), Salmonella spp (1.96%), Enterobacter sakazakii (1.96%), Serratia fonticola (1.96%). All E. coli showed total resistance to amoxicillin, ceftazidime, and cefotaxime. In addition, they showed variable resistance to norfloxacin (95.33%), trimethoprim (83.33%), and sulfamethoxazole (95.24%). 70.83% of E. coli strains isolated from apparently healthy children carry resistance genes blaTEM (12.5%), blaSHV (25%), and blaCTX-M (33.33%). While 66.67% of strains of E. coli isolated from children with diarrhea carry resistance genes blaTEM (12.5%), blaSHV (37.5%), and blaCTX-M (16.67%). Only EPEC was isolated and only in diarrheal children, with a frequency of 20.83%.
 Conclusion: The relatively high prevalence of ECD in this study is remarkable, and it highlights the fact that E. coli is an important agent of infectious diarrhea and could be the leading cause of gastroenteritis in Benin.

Antibiotic Resistance in Metal-Tolerant Microorganisms from Treatment Facilities.

The study examines the antibiotic resistance of metal-tolerant bacteria isolated from the wastewater treatment plant of a large city to six antibiotics belonging to the β-lactam antibiotics, aminoglycosides and amphenicols. Resistance of bacteria from sewage sludge multitolerant to heavy metals to 18 antibiotics of the β-lactam antibiotics, tetracyclines, aminoglycosides, diaminopyrimidines, amphenicols and ansamycins was studied also. Out of 10, the metal-tolerant microorganisms isolated from wastewater treatment facilities only the Klebsiella pneumonia strain (tolerant to 3 mM Cu) from the sludge of a secondary settling tank did not show resistance to the studied antibiotics at the concentrations considered. Resistance to the maximum amount of antibiotics was typical for strains Serratia fonticola SS0-1, isolated from fresh sewage sludge and resistant to 5 mmol Cu and 3 mmol Pb, or Stenotrophomonas maltophilia SS0-5, also isolated from fresh sludge and resistant to 3 mmol Zn and Cu. It is possible that bacterial resistance to antibiotics develops not only as a result of the use of antibiotics themselves, but also as a result of environmental pollution with heavy metals, and vice versa.

Evaluating of the Synergistic Effect of three Essential Oils on the Virulence Factors (resistance gene) of Pseudomonas aeruginosa and Serratia fonticola Isolated from Wounds

Depending on the high resistance to antibiotics, five isolates of Pseudomonas aeruginosa and 7 isolates of Serratia fonticola were selected out of 150 bacterial isolates from burn wards in Baghdad hospitals, which were later identified by VITEK2. A susceptibility test was done by using 15 antibiotics. The results showed that all the selected isolates were resistant to antibiotics: AMP, CTX, CAZ, GEN, PIP, TIC and TMP especially, while they were sensitive to IPE. The essential oils of Aloysia citrodora (Family: Verbenaceae), Rosmarinus officinalis (Family: Lamiaceae) and Thymus vulgaris (Family: Lamiaceae) were extracted by the Clevenger. The synergistic effect of essential oils of these plants on bacterial growth were studied by determining the MIC of essential oils for P. aeruginosa and S. fonticola. The MIC concentration of P. aeruginosa was: 2500 ppm of A. citrodora, 2500 ppm of R. officinalis, 2500 ppm of T. vulgaris, whilst the synergistic effect of each two plants was:1250 ppm of A. citrodora with 2500 ppm of R. officinalis, 1250 ppm of A. citrodora with 312.5 ppm of T. vulgaris, 1250 ppm of R. officinalis with 156.25 ppm of T. vulgaris, whereas the synergistic effect of three plants was: 156.25 ppm of A. citrodora with 156.25 ppm of R. officinalis and 39.0625 ppm T. vulgaris. While the MIC concentration of S. fonticola was: 1250 ppm of A. citrodora, 5000 ppm of R. officinalis, 1250 ppm of T. vulgaris, whilst the synergistic effect of each two plants was: 1250 ppm of A. citrodora with 2500 ppm of R. officinalis, 1250 ppm of A. citrodora with 312.5 ppm of T. vulgaris, 1250 ppm of R. officinalis with 156.25 ppm of T. vulgaris, whereas the synergistic effect of three plants was: 312.5 ppm of A. citrodora with 312.5 ppm of R. officinalis and 78.125 ppm T. vulgaris. The results of bacterial gene expression showed that three isolates of P. aeruginosa had a mexB gene expression that ranged between 2.5 - 0.4 (concentration 1) and 2.8 to 0.6 (concentration 2). It increased in two isolates and decreased in one isolate. The melting point ranged between 90.91 - 91.280C compared with 89.89 to 90.330C for fbp gene. Whereas S. fonticola did not have blaKPc gene, instead it had blaTEM gene with the gene expression ranging between 2.4 - 0.1 (concentration 1) and 4.5 - 0.2 (concentration 2). The gene expression increased in two isolates and decreased in five isolates. The melting point ranged between 83.63 - 84.200C compared with 75.82 - 85.560C for gyrB gene.

Bioremediation by bacteria isolated from water contaminated with hydrocarbons

Oil pollution is currently a global problem. However, an oil-contaminated ecology is rich in microorganisms that may utilize petroleum oil and hydrocarbons for growth, feeding, and metabolic processes. In the present study, fifty polluted water samples were collected from five stations (ten samples each) in the Al-Fahama oil refinery in eastern Baghdad. The water contamination parameters of these collected water samples were detected. Then, the percentage of water contamination with some heavy metals (zinc, lead, and cadmium) and radioactive elements (uranium, cesium and actinium) was measured. The proportions of these elements were compared within their limits permitted by the World Health Organization (WHO). Fifty-nine bacterial isolates were isolated from polluted water, and 24 isolates of them succeeded in analyzing crude oil. The results of the current study showed that seven isolates belong to the genus Citrobacter amalonaticus (29.16%), six isolates belong to Enterobacter cloacae (25%), three isolates belonged to both Pseudomonas aeruginosa (12.5%) and Ochrobacterum anthropi (12.5%), and human Ochrobacterum. With a percentage of 12.5%, two isolates of Serratia marcescens (8.3%) and one isolate of each Pseudomonas fluorescens, Serratia fonticola, and Burkholderia pseudomallei (4.16%) of each. The optimum of some conditions for the decomposition process was determined in terms of (pH, temperature and crude oil concentration) and the results showed that the optimum degradation conditions were 35°C at pH equal to 7.5 in the presence of 2% of crude oil. Several experiments were conducted to determine the most efficient isolates for oil analysis. Burkholderia pseudomallei and Pseudomonas fluorescens are the most active bacterial species in their oil degradation. Genes responsible for hydrocarbon analysis were revealed in twenty-four bacterial isolates using a polymerase chain reaction (PCR) assay. The results showed that the ALKB gene (alkane hydroxylase) was observed in all bacterial isolates that succeeded in analyzing crude oil with a percentage equal to 100%, NahAc gene (naphthalene dioxygenase) has been recorded in four isolates (16.7%), these four bacterial isolates were Burkholderia pseudomallei, Pseudomonas aeruginosa, Ochrobacterum anthropic, and Pseudomonas fluorescens. Generally, the isolation rate of both C. amalonaticus and E. cloacae isolates was higher than in other studies, which may be due to the hydrocarbon pollution in isolation; both B. pseudomallei and P. fluorescens isolates were the highest active bacterial species in their oil degradation. Genetic results showed that the AlkB gene was the domain compared with other degradation genes used in the current study, followed by NahAc gene. Keywords: Bioremediation, heavy metal, B. pseudomallei, hydrocarbons, crude oil

Engineered Gut Symbiotic Bacterium-Mediated RNAi for Effective Control of Anopheles Mosquito Larvae.

Anopheles mosquitoes are the primary vectors for the transmission of malaria parasites, which poses a devastating burden on global public health and welfare. The recent invasion of Anopheles stephensi in Africa has made malaria eradication more challenging due to its outdoor biting behavior and widespread resistance to insecticides. To address this issue, we developed a new approach for mosquito larvae control using gut microbiota-mediated RNA interference (RNAi). We engineered a mosquito symbiotic gut bacterium, Serratia fonticola, by deleting its RNase III gene to produce double-stranded RNAs (dsRNAs) in the mosquito larval gut. We found that the engineered S. fonticola strains can stably colonize mosquito larval guts and produce dsRNAs dsMet or dsEcR to activate RNAi and effectively suppress the expression of methoprene-tolerant gene Met and ecdysone receptor gene EcR, which encode receptors for juvenile hormone and ecdysone pathways in mosquitoes, respectively. Importantly, the engineered S. fonticola strains markedly inhibit the development of A. stephensi larvae and leads to a high mortality, providing an effective dsRNA delivery system for silencing genes in insects and a novel RNAi-mediated pest control strategy. Collectively, our symbiont-mediated RNAi (smRNAi) approach offers an innovative and sustainable method for controlling mosquito larvae and provides a promising strategy for combating malaria. IMPORTANCE Mosquitoes are vectors for various diseases, imposing a significant threat to public health globally. The recent invasion of A. stephensi in Africa has made malaria eradication more challenging due to its outdoor biting behavior and widespread resistance to insecticides. RNA interference (RNAi) is a promising approach that uses dsRNA to silence specific genes in pests. This study presents the use of a gut symbiotic bacterium, Serratia fonticola, as an efficient delivery system of dsRNA for RNAi-mediated pest control. The knockout of RNase III, a dsRNA-specific endonuclease gene, in S. fonticola using CRISPR-Cas9 led to efficient dsRNA production. Engineered strains of S. fonticola can colonize the mosquito larval gut and effectively suppress the expression of two critical genes, Met and EcR, which inhibit mosquito development and cause high mortality in mosquito larvae. This study highlights the potential of exploring the mosquito microbiota as a source of dsRNA for RNAi-based pest control.

Bioelectricity generation through Microbial Fuel Cells using Serratia fonticola bacteria and Rhodotorula glutinis yeast

Nowadays, there is great interest in microbial fuel cells because of the different substrates that can be used in them for electric energy generation. In order to find an alternative and contribute with eco-friendly technologies, this research used the Serratia fonticola bacteria and Rhotula glutinis yeast as a fuel source through laboratory scale microbial fuel cells. A single chamber microbial fuel cell with air cathode was fabricated with a copper foil and a graphite plate as anode and cathode electrode respectively. For the characterization of the cells, physicochemical parameters such as voltage, electric current, pH and electrical conductivity were measured for 30 days and at room temperature (18 ± 2.2 °C). It was possible to generate peak voltage and current values of 0.53 ± 0.01 V and 0.55 ± 0.02 V and current values of 1.76 ± 0.16 mA and 1.52 ± 0.02 mA, for MFCs with bacteria and yeast respectively. In addition, acidic operating pH was observed, and its conductivity peak values around 242 mS/cm. Finally, this work demonstrates the great potential that microorganisms have for the generation of electric current, giving a new and promising way to generate electricity

Quorum sensing inhibition through site-directed mutation by deletion PCR

Quorum sensing induces biofilms and virulence factors that are adverse industrially and medically. Nowadays, quorum sensing inhibitions focus on signal analogs or signal degradation, but these methods have several downsides, which are temporal and affected by several environmental factors. In this research, we used deletion PCR to perform site-directed mutagenesis on the quorum sensing pathway gene and then analyzed its effects on quorum sensing. Serratia fonticola DSM 4576 strain was utilized as the research strain, and the gram-negative bacteria's universal quorum sensing pathway, which is conducted by acyl-homoserine lactone (AHL), was analyzed. The structure and active site of the AHL synthase enzyme encoded by S. fonticola DSM 4576's luxI-type gene were predicted. The gene's partial section solely encodes the enzyme's active site. By using sequence and ligation-independent cloning, the obtained mutagenic gene was cloned into the suicide vector pEX18Ap. The recombinant vector was used to transform wild-type S. fonticola DSM 4576 strains, and the mutants were determined through two-step selections and PCR genotyping. The gene expression level and biofilm formation were quantitatively analyzed through RT-PCR and biofilm assay, and no significant difference was noted in the gene expression between wild types and mutants. However, when mutants were compared to wildtypes, there was a significant decrease in biofilm formation as a result of quorum sensing induced bioreaction. Thus, we propose a quorum sensing inhibitory technique based on enzyme mutation on the quorum sensing pathway, and we proved the feasibility of enzyme active site's site-directed mutation through deletion PCR.

Serratia silvae sp. nov., Isolated from Forest Soil.

The Gram-negative, oxidase-negative, catalase-positive, rod-shaped strain Arafor3T was isolated from forest soil (France). Comparative 16S rRNA gene analysis and phylogenetic analysis based on (1) multilocus sequence analysis (MLSA) with four housekeeping genes (atpD, gyrB, infB and rpoB) and (2) genomes indicated that strain Arafor3T shared 98.83% 16S rRNA gene sequence similarity with the type strain of Serratia fonticola DSM 4576T and was closely related to this same strain in the MLSA and in the phylogenomic tree reconstruction. Average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) comparisons of strain Arafor3T with its nearest neighbor S. fonticola DSM 4576T showed 93.5% identity and 55.7% sequence similarity, respectively, and were lower than the 96% and 70% species-level cut-off values relating to these analyses (Logan et al. in Int J Syst Evol Microbiol 59:2114-21, 2009, https://doi.org/10.1099/ijs.0.013649-0 ). The strain differed from S. fonticola in that it was urease and arginine dihydrolase negative. The major fatty acids of strain Arafor3T are C16:0, C16:1 ω7c/C16:1 ω6c, C14:0, C14:0 3-OH/16:1 isoI, and C18:1 ω7c. The major respiratory quinone is Q8. The polar lipid profile consisted of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, and 6 unknown lipids. The mol G + C% content of the genomic DNA of strain Arafor3T was 53.49%. Hence, Arafor3T represents a novel species within the genus Serratia, for which the name Serratia silvae sp. nov. is proposed. The type strain is Arafor3T (=LMG 32338T = CIP 111939T).