Novel in-genome based analysis demonstrates the evolution of OmpK37, antimicrobial resistance gene from a potentially pathogenic pandrug resistant Klebsiella pneumoniae MS1 isolated from healthy broiler feces.

Antimicrobial resistance transmission from farm animals to humans is a critical health concern and hence a detailed molecular surveillance is essential for tracking the spread and consequent evolution of antimicrobial resistance. In this study, a pan-drug resistant Klebsiella pneumoniae MS1 strain was isolated from a healthy broiler farm and studied. From the results of the study, MS1 was found to be is resistant to 18 tested antibiotics and has a high-risk to be pathogenic to humans with a probability of 0.80. The whole genome sequencing data of MS1 was used to predict the presence of antimicrobial resistance genes and pathogenicity. The genome analysis has revealed MS1 to have 34 AMR genes. Out of these, the AMR gene OmpK37 codes for an important protein involved in cell permeability and hence in antibiotic resistance. Further analysis was carried out by using an in-genome analysis method to understand the evolution of OmpK37 and the underlying reason for the emergence of resistance. From the detailed analysis, the current study could demonstrate for the first time the evolution of OmpK37 from OmpC. Though structurally OmpK37 was very similar to other porins present in MS1, it was found to have higher mutability as a distinguishing feature which makes it an important protein in monitoring the evolving resistances in microorganisms.

Predicting Human Risk with Multidrug Resistant Enterobacter hormaechei MS2 having MCR 9 Gene Isolated from the Feces of Healthy Broiler Through Whole-Genome Sequence-Based Analysis.

The zoonotic spread of antimicrobial resistance (AMR) and the associated infections are becoming a major threat to the human population worldwide. Strategies to identify the potential pathogen dissemination by seemingly healthy livestock are at a nascent stage and it is of significant importance to monitor environmental evolution of AMR. In this study, a multidrug resistant strain (MDR) of Enterobacter hormaechei MS2 isolated from the feces of healthy broiler chicken has been characterized by whole-genome sequencing-based method. Here, the isolate was primarily subjected to antimicrobial susceptibility testing followed genome sequencing and analysis. From the antimicrobial susceptibility testing result, the strain was found to be resistant to multiple classes of drugs including the colistin which is an important  last resort drug used to treat infectious diseases. The resistome prediction of genomic data further revealed the presence of 7 perfect and 26 strict hits including those for MCR-9 and FosA2. The pathogenicity prediction has also demonstrated the strain to have the potential to be a human pathogen with 0.72 probability. The phylogenetic analysis has also supported the zoonotic potential of the strain due to its clustering with isolates from both human and livestock-associated host groups. The results of the study suggest the need for a strong surveillance system to identify the opportunistic zoonotic pathogens to prevent a silent AMR menace mediated by them. Carriage of multi-drug resistant strains in the livestock gut microbiome is also a serious concern as it has high AMR transmissibility through contact and supply chain activities.

Bioactive Metabolites of Serratia sp. NhPB1 Isolated from Pitcher of Nepenthes and its Application to Control Pythium aphanidermatum.

Plant-associated bacteria have already been considered as the store house of bioactive compounds that confer the plant growth promotion and disease protection. Hence, the unique plant parts have already been expected to harbor diverse microbial communities with multi-beneficial properties. Based on this, the current study has been designed to identify the potential of Serratia sp. NhPB1 isolated from the pitcher of Nepenthes plant for its activity against the infamous pathogen Pythium aphanidermatum. The in vitro antifungal, plant growth promoting and enzymatic activities of the isolate indicated its promises for agricultural application. The isolate NhPB1 was also demonstrated to have positive effect on Solanum lycopersicum and Capsicum annuum, due to its plant beneficial metabolites. From the results of LC-MS/MS analysis, the isolate has also been revealed to have the ability to synthesize bioactive compounds including salicylic acid, cyclodipeptides, acyl homoserine lactone, indole-3-acetic acid, and serrawettin W1. These identified compounds and their known biological properties make the isolate characterized in the study to have significant promises as an eco-friendly solution for the improvement of agricultural productivity.

Prevalence of Antimicrobial Resistance Among the Hydrogen Sulfide Producing Bacteria Isolated on XLD Agar from the Poultry Fecal Samples.

Poultry products remain as one of the most popular and extensively consumed foods in the world and the introduction of hydrogen sulfide (H2S) producing antibiotic resistant bacterial species into it is an emerging challenge. The current study has been designed to analyze the distribution of antibiotic resistance among the H2S producing bacteria isolated from the fecal samples of chickens from different poultry farms. Here, twenty bacterial isolates were selected based on their ability to produce H2S on XLD agar, and the16S rDNA sequencing was carried out for their molecular identification. The results showed the isolates as belong to Salmonella spp. and Citrobacter spp. and in the antibiotic susceptibility test (AST), three of the Salmonella strains were found to be resistant to antibiotics such as tetracycline, doxycycline, nalidixic acid, and amikacin. Also, fourteen Citrobacter strains showed resistance towards azithromycin, and furthermore, eleven of them were also resistant to streptomycin. Resistance towards tetracycline was observed among five of the Citrobacter strains, and seven were resistant to doxycycline. Further molecular screening by the PCR has showed three of the Salmonella strains along with eight Citrobacter isolates to have tetA gene along with four of the Citrobacter strains to have co-harbored blaTEM gene. The results on biofilm formation have also demonstrated three Salmonella strains along with nine Citrobacter strains to have the ability to form moderate biofilm. The study thus describes the occurrence of H2S producing multidrug-resistant bacteria in poultry feces, which might contribute towards the dissemination of antibiotic resistance genes to other microorganisms including human pathogens with likely risk to treat disease conditions.

9-Tricosene Containing Blend of Volatiles Produced by Serratia sp. NhPB1 Isolated from the Pitcher Plant Provide Plant Protection Against Pythium aphanidermatum.

Plant-associated bacteria exhibit diverse chemical means to protect plants from the pathogens. The present study has been conducted to evaluate the volatile-mediated antifungal activity of Serratia sp. NhPB1 isolated from the pitcher plant against the notorious pathogen Pythium aphanidermatum. The study has also evaluated the protective effect of NhPB1 on Solanum lycopersicum and Capsicum annuum leaves and fruits against P. aphanidermatum. From the results, NhPB1 was found to have remarkable activity against the tested pathogen. The isolate was also found to impart disease protection in selected plants as evidenced by the morphological changes. Here, the leaves and fruits of S. lycopersicum and C. annuum control which were treated with the uninoculated LB and distilled water were found to have the presence of P. aphanidermatum growth with lesions and decaying of tissues. However, the NhPB1-treated plants did not show any symptoms of fungal infection. This could further be confirmed by the microscopical examination of tissues by propidium iodide staining. Here, the normal architecture of leaf and fruit tissues could be observed in the NhPB1-treated group, but the tissue invasion by P. aphanidermatum was observed in the control group which further confirms the promises of selected bacteria for biocontrol applications.

Multifarious Plant Probiotic Features of Bacillus sp. W11 Isolated from Vermicast and Its Promises for Biocontrol Activity Against Phytopathogens.

Organic agricultural practice using vermisources is considered as one of the common sustainable strategies in agriculture. Apart from their nutrient content, beneficial microbes associated with natural vermicast serve as an efficient bioinoculant for improving agricultural productivity. Hence, studies on the identification of suitable microbial inoculants with multi-trait plant beneficial properties from these hotspots are highly promising as an ecofriendly substitute against harmful chemical fertilizers. The current study has been designed to isolate bacterial strains with various plant probiotic and biocontrol properties against various fungal phytopathogens. Among the various bacteria obtained in the study, the isolate W11 was found to have remarkable inhibitory activity against a broad range of phytopathogens like Fusarium oxysporum, Pythium myriotylum, and Rhizoctonia solani, in addition to its multiple plant growth-promoting properties. The W11 was further identified as Bacillus sp. by 16S rDNA sequence-based analysis. The W11 treatment was also found to enhance the plant growth parameters of Vigna unguiculata. In addition, the priming of Bacillus sp. W11 on potato tuber has confirmed to protect it from Fusarium wilt caused by Fusarium oxysporum. This highlights the possible protective effect of W11 during the post-harvest storage of potatoes. Also, the metabolite analysis of W11 extract by LC-QTOF-MS/MS analysis has revealed the presence of lipopeptide surfactin derivatives with m/z of 1008, 1022, and 1036 (M + H)+. All the results obtained in the study thus indicate the remarkable agricultural promises of Bacillus sp. W11 isolated from vermicast. Even though vermicast has been studied for its beneficial agricultural applications, the isolation of plant beneficial bacteria from it and detailed characterization of its beneficial effects make the study important.

Spectrophotometric Evaluation of Translucency of Various Commercially Available Zirconium Oxide Ceramic Systems: An In Vitro Study.

AIM: To evaluate and compare the translucency of various commercially available zirconium oxide ceramic systems, i.e., Ceramill® Zolid Classic, Ceramill® Zi, and DD Bio ZX²71 by using a dual beam UV-visible spectrophotometer. MATERIALS AND METHODS: The present study comprised of 21 disk-shaped samples of zirconia for every group, i.e., group I-Ceramill® Zolid Classic, group II-Ceramill Zi®, and group III-DD Bio ZX²71. Furthermore, each group was split into three subgroups and every subgroup had seven samples each one of 0.7, 0.8, and 0.9 mm thickness. The samples were prepared by computer-aided design/computer-aided manufacturing (CAD/CAM) system devised by Amann Girrbach AG in accord with the steps provided by the manufacturer. The entire sample was designed having 10 mm in diameter with 0.7, 0.8, and 0.9 mm thickness for every group. The UV-visible dual beam spectrophotometer equipped with D2 lamp and W lamp was used for the measurement of absorbance and transmittance in order to assess the translucency of the fabricated zirconia samples. RESULTS: The mean value of transmittance % for Ceramill® Zi at 0.8 mm came out to be 0.849 ± 0.024, i.e., the least among all, whereas the mean value of Ceramill® Zolid Classic was 1.408 ± 0.033, being the highest for the same thickness. DD Bio ZX²71 had an intermediate value of 1.274 ± 0.012. The mean value of absorbance for Ceramill® Zi at 0.8 mm came out to be 2.086 ± 0.013, i.e., the maximum among all, whereas the mean value of Ceramill® Zolid Classic was, being the lowest for the same thickness. DD Bio ZX²71 had an intermediate value of 1.902 ± 0.004. CONCLUSION: The present study data suggest that all the materials subjected to evaluation exhibited a substantial translucency. We attempted to study few of the desirable properties, these materials should possess when used for prosthetic rehabilitation with esthetic contentment a clinical setup. There has been an ambiguous distinction that Ceramill® Zi Zirconia supersedes the Ceramill® Zolid Classic and DD Bio ZX²71. Furthermore, 0.8 mm thickness substantiates to be the most ideal among 0.7, 0.8, and 0.9 mm. CLINICAL SIGNIFICANCE: The desired outcome of the procedure becomes dependent solely on the clinician's judgment to opt for the material whose properties are most fitting as per the demands of the esthetics. While a clinician should always be ambitious, but a good clinician should also bear in mind that the success of any treatment procedure not only depends on the assortment of properties of these materials but also the host response and satisfaction evoked by these materials.

Rapid detection of mobile resistance genes tetA and tetB from metaplasmid isolated from healthy broiler feces.

Containing antimicrobial resistance is the thought of the moment as it affects the human life from every aspect. Because, the inappropriate use of antibiotics in livestock animals for the growth promotion and prophylactic purpose has already generated significant challenges. The livestock farms which harbor and disseminate drug resistant microorganisms have already been identified as potential source of resistance genes acquired by the sensitive strains. Hence there is high demand for the affordable and effective surveillance method for the detection of antimicrobial resistance genes from livestock. In this study, direct detection of antibiotic resistance from metaplasmid DNA isolated from the poultry feces was conducted. For the initial standardization, plasmid DNA purified from the previously characterized Escherichia coli and Klebsiella pneumoniae were used. The tetA and tetB genes amplified from the purified plasmid DNA were further confirmed by agarose gel electrophoresis and sequencing. Further to this, metaplasmid DNA was purified from 29 different poultry fecal samples and these were further screened for the presence of resistance genes. Among the 29 metaplasmid samples, 8 were positive for tetA gene and 9 were positive for tetB gene. The results of the study indicate the potential of PCR based methods for the rapid screening of poultry samples for the antibiotic stewardship in the livestock sector.

Protective effect of (+)-catechin against lipopolysaccharide-induced inflammatory response in RAW 264.7 cells through downregulation of NF-κB and p38 MAPK.

Catechin, a flavonol belonging to the flavonoid group of polyphenols is present in many plant foods. The present study was done to evaluate the effect of catechin on various inflammatory mediators using lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. The effect of catechin on total cyclooxygenase (COX) activity, 5-lipoxygenase (5-LOX), myeloperoxidase, nitrite and inducible nitric oxide synthase (iNOS) level, secretion of tumor necrosis factor-α (TNF-α) and interleukin-10 (IL-10) were assessed in LPS-stimulated RAW 264.7 cells. The expression of COX-2, iNOS, TNF-α, nuclear factor-ĸB (NF-κB) and p38 mitogen-activated protein kinase (MAPK) genes were also investigated. The effect was further analyzed using human PBMCs by assessing the level of TNF-α and IL-10. The study demonstrated that the inflammatory mediators such as COX, 5-LOX, nitrite, iNOS, and TNF-α were significantly inhibited by catechin in a concentration-dependent manner whereas IL-10 production was up-regulated in RAW 264.7 cells. Moreover, catechin down-regulated the mRNA level expression of COX-2, iNOS, TNF-α, NF-κB and p38 MAPK. The current study ratifies the beneficial effect of catechin as a dietary component in plant foods to provide protection against inflammatory diseases.

Genome sequencing and annotation of multi-virulent Aeromonas veronii XhG1.2 isolated from diseased Xiphophorus hellerii.

The present study was intended to elucidate the genomic basis of antibiotic resistance and hyper-virulence of the fish pathogen Aeromonas veronii XhG1.2 characterized in our previous work. The identity of XhG1.2 was confirmed through 16S rDNA sequence analysis and whole genome sequence analysis. The top-hit species distribution analysis of XhG1.2 sequence data revealed major hits against the Aeromonas veronii. The identification of virulence genes using the VFDB showed the genome of XhG1.2 to have the genes coding for the virulence factors viz. aerolysin, RtxA, T2SS, T3SS and T6SS. The presence of antibiotic resistance predicted through the CARD database analysis showed it to have the CephA3, OXA-12, adeF and pulvomycin resistance genes. By the phylogenetic and comparative genomic analysis, A. veronii species were found to have genes for toxin production. This also confirmed the pathogenicity and drug resistance of A. veronii XhG1.2 and also its potential to cause disease in diverse ornamental fishes.

Healthy broilers disseminate antibiotic resistance in response to tetracycline input in feed concentrates.

Wide varieties of antibiotics are used in poultry farms to improve the growth and also to control the infection in broiler chicken. To identify the seriousness of the same in the poultry sector, current study has been designed to analyze the presence of tetracycline in poultry feed and also the tetracycline resistance among the bacteria released through the excreta of poultry. In the study, 27 bacteria belonging to the Escherichiacoli and Klebsiellapneumoniae. were isolated from the faecal samples collected from five different farms. Antibiotic susceptibility analysis showed 77% of E. coli and 100% of the K. pneumoniae. to be resistant to tetracycline. Further, molecular screening for tetA and tetB genes showed 85.18% of isolates to have tetA and 22.22% with tetB. The presence of tetracycline in collected feed samples was also analysed quantitatively by Liquid chromatography-mass spectrometry (LC-MS). Here, three out of five feed samples were found to be positive for tetracycline. The study showed a direct correlation between the antibiotic supplemented feed and the emergence of antimicrobial resistance among the intestinal microflora. The results of the study indicate the need for strict control over antibiotic use in animal feed to limit the rapid evolution and spread of antimicrobial resistance.

Advantage of zinc oxide nanoparticles over silver nanoparticles for the management of Aeromonas veronii infection in Xiphophorus hellerii.

Bacterial pathogens cause significant challenge to the ornamental fish industry. Eventhough antibiotic administration has been recommended to manage fish diseases, there is alarming concern with emergence of antibiotic resistance. This indicates the need for the development of alternative methods with multi-targeted action to manage fish diseases. In the study, silver (AgNPs) and zinc oxide (ZnONPs) nanoparticles have been evaluated for its activity against Aeromonas veronii. Both the AgNPs and ZnONPs were found to have bactericidal activity against A. veronii. In vivo experiments with A. veronii was found to cause severe mortality in Xiphophorus hellerii with a LD50 of 1.4 × 108 CFU/mL. However, treatment with AgNPs and ZnONPs each at a concentration of 1 mg/L was found to enhance the survival rate of X. hellerii to 83.3% and 100% respectively. Further histopathological analysis showed alterations in gill, intestine and liver of X. hellerii due to A. veronii in the infection control. In the case of AgNPs treated group, symptoms of moderate tissue damage could be observed. However, the ZnONPs treated X. hellerii showed normal histological features with minimum tissue damage. The bath dip method further confirmed the protective ability of the zinc oxide nanoparticles (1 mg/L) on X. hellerii.

Antibacterial Potential and Apoptosis Induction by Pigments from the Endophyte Burkholderia sp. WYAT7.

Pigment from the endophyte Burkholderia sp. WYAT7 isolated from the medicinal plant Artemisia nilagirica (Clarke) Pamp. was extracted. The antibacterial efficacy of the crude pigment Y was assessed as a source of antibiotic against both Gram-positive and Gram-negative bacterial pathogens. The pigment Y exhibited a significant level of antibacterial activity against the tested pathogens Salmonella typhi (MTCC 733), Staphylococcus aureus (MTCC 1430), Pseudomonas aeruginosa (MTCC 2453), Klebsiella pneumoniae (MTCC 432), Escherichia coli (MTCC 1610), Salmonella paratyphi (3220), Bacillus subtilis (441) and Acinetobacter baumannii (12,889). The minimum inhibitory concentration of crude pigment extract Y for most of the bacterial pathogens tested was below or equal to 0.25 µg/mL and the minimum bactericidal concentration was below or equal to 0.5 µg/mL. In the cytotoxicity evaluation, crude pigment Y exhibited less toxicity toward normal cells lines (L929). Crude pigment extract Y also showed powerful anticancer activity toward melanoma cancer cells (A375). The IC50 value obtained was 68.08 µg/mL. Acridine orange (AO) and ethidium bromide (EB) double staining of cells treated with the pigment helped in the morphological assessment of nuclear condensation, apoptotic bodies and live cells. The DNA fragmentation analysis and caspase-9 quantification in the pigment-treated A375 cells determined the apoptosis activity mediated by the crude pigment extract Y. The compounds in the crude pigment extract Y was identified by HR-LCMS analysis. Further studies on the active compounds can lead to a rise in new drugs for cancer treatment and also against rising antibacterial resistant pathogens.

Characterization of biosurfactant produced by the endophyte Burkholderia sp. WYAT7 and evaluation of its antibacterial and antibiofilm potentials.

The endophyte Burkholderia sp. WYAT7 isolated from the medicinal plant Artemisia nilagirica (Clarke) Pamp. was analyzed for its ability to produce biosurfactant. The evaluation of biosurfactant production was conducted using different screening methods which confirmed the presence of biosurfactant in the culture supernatant. CTAB- methylene blue agar plate method was used for the screening of glycolipid biosurfactant production. The biosurfactant produced by the bacteria effectively metabolized hydrocarbons present in the bacterial culture media. Fourier transform infrared spectroscopic (FTIR) analysis of biosurfactant provided the details regarding OH stretching, stretching vibrations of acyl chain, CO stretching, stretching vibrations of ether and vibrations of glycosidic linkages in the biosurfactant. The stretching vibrations of glycosidic linkage in the fingerprint regions of FTIR spectrum (1200 cm-1 to 800 cm-1 regions) confirms that the biosurfactant produced was a glycolipid. The GC-MS analysis confirmed the methyl and ethyl esters of fatty acids. The biosurfactant from the bacteria exhibited antibacterial activity against bacterial pathogens such as Pseudomonas aeruginosa (MTCC 2453), Escherichia coli (MTCC 1610), Salmonella paratyphi and Bacillus subtilis. The glycolipid biosurfactant had antibiofilm activity as evidenced in Staphylococcus aureus (MTCC 1430). All these results indicated the beneficial effect of the biosurfactant in plant-endophyte interactions. The properties exhibited by the biosurfactant suggest that it can be exploited commercially for the production of novel antibiotics.

Rotula aquatica Lour. inhibits growth and biofilm formation of clinically isolated uropathogenic Escherichia coli

Objective: To evaluate the anti-bacteria! and anti-biofilm activity of ethyl acetate fraction of Rotula aquatica Lour. (EFRA) against clinically isolated uropathogenic Escherichia coli. Methods: In vitro antibacterial and anti-biofilm studies were employed. The antimicrobial activity of EFRA was assayed by the well diffusion method. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the active fraction were determined by Resazurin method. The time-kill kinetic assay, acridine orange-ethidium bromide staining, propidium iodide uptake assay, and scanning electron microscopic (SEM) analysis were done to evaluate the efficacy of EFRA in killing uropathogenic Escherichia coli. The anti-biofilm activity was determined by 3-[4,5- dimethyl-2-thiazolyl]-2, 5-diphenyl-2H-tetrazolium-bromide (MTT) assay and specific biofilm formation assay. Results: The well diffusion assay of EFRA showed a very clear zone of inhibition against Escherichia coli BRL-17. The MIC and MBC of EFRA were 2.5 mg/mL and 5 mg/mL, respectively. The time-kill kinetic assay, fluorescence microscopic analysis, propidium iodide uptake assay, and SEM analysis displayed the effect of EFRA in killing the bacteria. The MTT assay and specific biofilm formation assay showed that EFRA prevented the formation of biofilms. Conclusions: The results of the present study confirm that EFRA could prevent bacterial growth and inhibit its biofilm formation.

Bacterial endophytes from Artemisia nilagirica (Clarke) Pamp., with antibacterial efficacy against human pathogens.

A study was conducted to isolate and characterize endophytes from Artemisia nilagirica, a traditional medicinal plant. The plant was collected from Western Ghats, India. Endophytes isolated included Arthrobacter sp. WWAT1, Pseudomonas sp. WYAT2, Microbacterium sp. WYAT3, Psychrobacter sp. WBAT4, Enterobacter sp. WWAT5, Bacillus sp. WBAT6, Kosakonia cowanii WBAT7, Bacillus sp. WBAT8, Bacillus sp. WBAT9, Chromobacterium violaceum WVAT6, Serratia sp.WPAT8 and Burkholderia sp. WYAT7. Of these two bacteria, Chromobacterium violaceum strain WVAT6 and Burkholderia sp. strain WYAT exhibited antibacterial property against human pathogens. Similar to the environmental isolates, Burkholderia sp. WYAT7 showed pleomorphism and produced different enzymes, whereas like clinical strains they showed multidrug resistance, for their survival in different environmental conditions. Chromobacterium violaceum WVAT6 exhibited rod shape morphology and showed multiple drug resistance except to erythromycin, tetracycline and gentamicin antibiotics. Both produced biofilm and enzymes such as protease and lipase. The antimicrobial compounds from these endophytes may find application in the preparation of antimicrobial formulations.

One-step synthesis of eco-friendly boiled rice starch blended polyvinyl alcohol bionanocomposite films decorated with in situ generated silver nanoparticles for food packaging purpose.

The study reports a one-step preparation of polyvinyl alcohol/boiled rice starch blend film fabricated with in situ generated silver nanoparticles (PVA/BRS/sAgNPs) formed in the presence of sunlight irradiation. The bionanocomposite appeared to have dark brown color with a characteristic surface plasmon resonance peak at 439 nm. Further characterization has confirmed the presence of physical interactions among the components PVA, BRS and sAgNPs. Compared to the control PVA, the nanocomposites showed improved mechanical and optical properties with decreased water sensitivity. Presence of boiled rice starch and sAgNPs were also found to influence the light transmittance of composite film. Moreover, PVA/BRS/sAgNPs film was found to have superior barrier property against environmental microorganisms. Biodegradation of the composite films was studied by indoor soil burial test and was assessed by visual appearance, weight loss and FTIR analysis. Interestingly, both the PVA/BRS and PVA/BRS/sAgNPs films proved to be biodegradable and hence have promising application as cost effective food packaging material with the latter having marked antimicrobial property.

Distribution of antibiotic resistance and virulence factors among the bacteria isolated from diseased Etroplus suratensis.

Considering the emerging concern with the antimicrobial resistance (AMR) evolution, the study has been designed to identify the antibiotic resistance and virulence properties of culturable bacteria isolated from the diseased fish Etroplus suratensis. This has resulted in the purification of 18 morphologically distinct bacterial isolates which were identified by both biochemical and molecular methods. Antibiotic resistance analysis showed the resistance of these isolates to multiple antibiotics and remarkable evolution of AMR. Further screening for virulence factors confirmed five isolates to be positive for haemolytic activity, eight with caseinase, four with DNase, one with gelatinase and three with biofilm-forming properties. In addition to these, the isolates were subjected to PCR-based screening to detect the presence of genes coding for aerolysin and haemolysin. Results showed the presence aerolysin gene in the isolates ESS3.2, ESS3.8, ESI3.3 and ESS3.6, while haemolysin gene was observed to be present in ESG3.1 and ESI3.2. The observed results hence indicate the need for frequent monitoring of these properties among bacterial isolates from diverse environment especially those associated with edible fish.

Assessment of Plant-Probiotic Performance of Novel Endophytic Bacillus sp. in Talc-Based Formulation.

Endophytic bacteria are considered to have a plethora of plant growth promoting and anti-phytopathogenic traits to live within the plants. Hence, they have immense promises for plant probiotic development. In the current study, plant probiotic endophytic Bacillus sp. CaB5 which has been previously isolated from Capsicum annuum was investigated for its performance in talc-based formulation. For this, CaB5 was made into formulation with sterile talc, calcium carbonate, and carboxymethyl cellulose. The viability analysis of the formulation by standard plate count and fluorescence methods has confirmed the stable microbial count up to 45 days. Plant probiotic performance of the prepared formulation was analyzed on cowpea (Vigna unguiculata) and lady's finger (Abelmoschus esculentus). The results showed the formulation treatment to have enhancement effect on seed germination as well as plant growth in both selected plants. The results highlight the potential of CaB5-based formulation for field application to enhance growth of economically important plants.