Probiotic Paenibacillus polymyxa HGA4C and Bacillus licheniformis HGA8B combination improved growth performance, enzymatic profile, gene expression and disease resistance in Oreochromisniloticus.

Bacterial consortium containing two bacterial strains such as Paenibacillus polymyxa HGA4C and Bacillus licheniformis HGA8B incorporated in the diet of Oreochromis niloticus at a concentration of 1 × 106 CFU g-1 (PB1) and 1 × 108 CFU g-1 (PB2) revealed the probiotic potentials of the bacterial combination. The probiotic feed enhanced the growth performance, digestive enzymes, and antioxidant enzymes in the liver and intestine. Probiotic mediated growth enhancement was further substantiated by the up-regulation of genes such as GHR-1, GHR-2, IGF-1, and IGF-2 and the up-regulation of immune-related genes viz. TLR-2, IL-10, and TNF-α were also significantly modulated by probiotics supplementation. The intestinal MUC 2 gene expression revealed the mucosal remodification and the disease resistance of the fish challenged with Aeromonas hydrophila (MTCC-1739) was improved by the probiotic supplementation. Based on these results the new probiotic supplementation feed can be possibly marketed to help aquaculture farmers to alleviate many of the problems associated with fish farming.

Rotula aquatica Lour. mitigates oxidative stress and inflammation in acute pyelonephritic rats.

The current study evaluates the efficacy of methanolic extract of Rotula aquatica Lour. (MERA) against inflammatory changes associated with acute pyelonephritis. The antioxidant enzymes such as SOD, CAT, GPx, GR and oxidative stress markers like GSH content, malondialdehyde (MDA) level, nitrate level, reactive oxygen species (ROS) level and renal toxicity markers were evaluated in this study. The mRNA level expression of Toll-like receptor 4 (TLR-4), nuclear transcription factor kappa B (NF-κB), tumour necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and Tamm Horsfall protein (THP) were studied by RT-PCR analysis. The oral administration of MERA increases the antioxidant enzyme status in pyelonephritis rat. The elevated levels of oxidative stress markers in pyelonephritic rats were ameliorated by the administration of MERA at 100 mg/kg and 200 mg/kg bwt of the rat. The mRNA level expression of major genes were restored to normal level by MERA.

Valencene-rich fraction from Vetiveria zizanioides exerts immunostimulatory effects in vitro and in mice

Objective: To decipher the responsible compound present in the aqueous root extract of Vetiveria zizanioides which has tremendous immunomodulatory activity. Methods: Different fractions of the water extract were collected and analyzed for immunomodulatory activity by analyzing in vitro phagocytic activity and nitric oxide production. One fraction VF3 was selected and further analyzed for possible compounds by high performance liquid chromatography and gas chromatography coupled with a mass spectrometer. The in vitro immunomodulatory parameters such as phagocytic index, nitrite content, and tumor necrosis factor-α production in murine macrophages were analyzed. In vivo studies, sheep red blood cell induced haemagglutination titer, the number of antibody-producing cells, and sheep red blood cell induced delayed-type hypersensitivity were analyzed. Cytotoxic studies in L929 normal fibroblasts were also performed. Results: One of the fractions, VF3, was selected and confirmed the presence of an active compound valencene. The in vitro immunomodulatory parameters were significantly (P<0.05) increased by valencene treatment. In vivo studies in Swiss albino mice showed that valencene could significantly (P<0.05) increase haemagglutination titer, the number of antibody-producing cells, and delayed-type hypersensitivity. Cytotoxic studies also showed that valencene did not cause any morphological changes and DNA damage in normal fibroblasts. Conclusions: Valencene possesses immunomodulatory activities and can be commercially exploited for its immunostimulatory potentials.

An in vitro and in silico study of anti-dermatophytic activity of gossypol from fruits of Thespesia populnea (L.) Sol. ex Correa

Objective: To isolate, purify, and characterize gossypol from the fruits of Thespesia populnea (L) Sol. ex Correa, test its anti- dermatophytic activity, identify its targets on the dermatophyte, and confirm the binding of gossypol with the fungal target by molecular docking study. Methods: Gossypol from Thespesia populnea was characterized by high performance liquid chromatography, liquid chromatograph- mass spectrometry, Fourier transform infrared spectroscopy, and nuclear magnetic resonance. The anti-dermatophytic activity of gossypol was tested against four different dermatophytes, viz. Trichophyton mentagrophytes, Trichophyton rubrum, Microsporum canis, and Microsporum gypseum. Trichophyton mentagrophytes was selected for further studies. The inhibitory mode of action of gossypol on Trichophyton mentagrophytes was determined by analyzing the modulation of gene expression in various pathways of the dermatophyte. Results: Gossypol inhibited all the dermatophytes. The minimum inhibitory concentrations were 12.5 μg/mL for Trichophyton mentagrophytes and Microsporum canis and 25 μg/mL for Trichophyton rubrum and Microsporum gypseum. The minimum fungicidal concentrations were 50 μg/mL for Trichophyton mentagrophytes, 100 μg/mL for Microsporum canis and Trichophyton rubrum, and 200 μg/mL for Microsporum gypseum. Gossypol inhibited the mRNA expression of metalloprotease (MEP4) and isocitrate lyase (ICL). The binding of gossypol with the enzymes was confirmed by molecular docking studies. The best docking poses were found and the low binding energies were recorded with the two target enzymes. Conclusions: Gossypol is a potential antifungal agent and can be further explored as an anti-dermatophytic drug.

Valencene-rich fraction from Vetiveria zizanioides exerts immunostimulatory effects in vitro and in mice

Objective: To decipher the responsible compound present in the aqueous root extract of Vetiveria zizanioides which has tremendous immunomodulatory activity. Methods: Different fractions of the water extract were collected and analyzed for immunomodulatory activity by analyzing in vitro phagocytic activity and nitric oxide production. One fraction VF3 was selected and further analyzed for possible compounds by high performance liquid chromatography and gas chromatography coupled with a mass spectrometer. The in vitro immunomodulatory parameters such as phagocytic index, nitrite content, and tumor necrosis factor-α production in murine macrophages were analyzed. In vivo studies, sheep red blood cell induced haemagglutination titer, the number of antibody-producing cells, and sheep red blood cell induced delayed-type hypersensitivity were analyzed. Cytotoxic studies in L929 normal fibroblasts were also performed. Results: One of the fractions, VF3, was selected and confirmed the presence of an active compound valencene. The in vitro immunomodulatory parameters were significantly (P<0.05) increased by valencene treatment. In vivo studies in Swiss albino mice showed that valencene could significantly (P<0.05) increase haemagglutination titer, the number of antibody-producing cells, and delayed-type hypersensitivity. Cytotoxic studies also showed that valencene did not cause any morphological changes and DNA damage in normal fibroblasts. Conclusions: Valencene possesses immunomodulatory activities and can be commercially exploited for its immunostimulatory potentials.

An in vitro and in silico study of anti-dermatophytic activity of gossypol from fruits of Thespesia populnea (L.) Sol. ex Correa

Objective: To isolate, purify, and characterize gossypol from the fruits of Thespesia populnea (L) Sol. ex Correa, test its anti- dermatophytic activity, identify its targets on the dermatophyte, and confirm the binding of gossypol with the fungal target by molecular docking study. Methods: Gossypol from Thespesia populnea was characterized by high performance liquid chromatography, liquid chromatograph- mass spectrometry, Fourier transform infrared spectroscopy, and nuclear magnetic resonance. The anti-dermatophytic activity of gossypol was tested against four different dermatophytes, viz. Trichophyton mentagrophytes, Trichophyton rubrum, Microsporum canis, and Microsporum gypseum. Trichophyton mentagrophytes was selected for further studies. The inhibitory mode of action of gossypol on Trichophyton mentagrophytes was determined by analyzing the modulation of gene expression in various pathways of the dermatophyte. Results: Gossypol inhibited all the dermatophytes. The minimum inhibitory concentrations were 12.5 μg/mL for Trichophyton mentagrophytes and Microsporum canis and 25 μg/mL for Trichophyton rubrum and Microsporum gypseum. The minimum fungicidal concentrations were 50 μg/mL for Trichophyton mentagrophytes, 100 μg/mL for Microsporum canis and Trichophyton rubrum, and 200 μg/mL for Microsporum gypseum. Gossypol inhibited the mRNA expression of metalloprotease (MEP4) and isocitrate lyase (ICL). The binding of gossypol with the enzymes was confirmed by molecular docking studies. The best docking poses were found and the low binding energies were recorded with the two target enzymes. Conclusions: Gossypol is a potential antifungal agent and can be further explored as an anti-dermatophytic drug.

Acute oral toxicity and anti-inflammatory evaluation of methanolic extract of Rotula aquatica roots in Wistar rats.

BACKGROUND: The plant Rotula aquatica Lour. was traditionally well known due to its large number of pharmacological action and medicinal uses. The plant is a necessary component of many Ayurvedic drug preparations since historical times. It is widely used as a crucial ancient drug for kidney and bladder stones. OBJECTIVES: The main objective of the study was to evaluate the acute toxicity and anti inflammatory efficacy of methanolic extract of R.aquatica Lour. in in vivo models. MATERIALS AND METHODS: The qualitative phytochemical analysis and invitro antioxidant activity of the roots of methanolic extract of R.aquatica Lour. (MERA) was evaluated. The acute toxicity effect of MERA was evaluated with two different doses (550, 2000 mg/kg body weight), were administrated orally to Wistar rats. The rats were observed for sign and symptoms of toxicity and mortality for 14 days. The parameters measured including relative organ weight, blood, biochemical and histopathological parameters of hepatic and renal toxicity. The anti-inflammatory effect of MERA was also evaluated in carrageenan and dextran-induced paw edema models. RESULTS: The phytochemical evaluation of MERA shows the presence of secondary metabolites like alkaloids, flavonoids, phenolics and tannins, phytosterols, reducing sugars, proteins and terpenoids. The results of in-vitro antioxidant evaluation of MERA reveal its capability to scavenging free radical at a lower concentration. The MERA did not show any visible signs of toxicity up to the dose of 2000 mg/kg body weight. The results obtained from our carrageenan and dextran-induced paw edema model study also proved the anti-inflammatory effect of MERA in rat model. CONCLUSION: The result shows the potential of MERA as an anti-inflammatory drug to reduce the signs of inflammation devoid of any toxic effect.

Surface functionalization of central venous catheter with mycofabricated silver nanoparticles and its antibiofilm activity on multidrug resistant Acinetobacter baumannii.

The mycofabrication of silver nanoparticles (AgNPs) through green chemistry protocol is an advanced methodological progress in medical nanotechnology. Mycofabricated AgNPs are less toxic due to an aura of biomolecules around the nanoparticles. Hence the mycofabricated AgNPs can be used for clinical applications. The present study explores the antibiofilm activity of mycogenerated AgNPs, which were synthesized by the enzymatic reduction of silver nitrate using the marine algicolous endophytic fungus Penicillium polonicum ARA10. The mycogenerated AgNPs showed very specific and potent bactericidal activity against Acinetobacter baumannii. Anti-A.baumannii activities of mycogenerated AgNPs on planktonic as well as biofilm embedded cells were explored. The physical impact of synthesized AgNPs on A.baumannii was investigated by scanning electron microscopy and fluorescence microscopy. A bionanocomposite coating for the central venous catheter (CVC) was formulated using the mycogenerated AgNPs and polydopamine. The bionanocomposite surface was characterized by field emission scanning electron microscopy, water contact angle measurement, and Raman spectroscopy. The results showed that the mycogenerated AgNPs have potent antibiofilm activity on biofilms of A.baumannii. The scanning electron microscopy (SEM) and fluorescence microscopy images showed noticeable aberrations on the ultrastructure of A.baumannii. The SEM and FE-SEM images of biofilms on the surface of CVC samples proved that the AgNPs at minimum bactericidal concentration could destroy the structure of biofilms and lyses the bacterial cell. Thus, the present study establishes a new way to the development of 'antibacterial surfaces' based on mycogenerated AgNPs.

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

Objective: To evaluate the anti-bacterial 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.

Catechin rich butanol fraction extracted from Acacia catechu L. (a thirst quencher) exhibits immunostimulatory potential.

Acacia catechu L., (Fabaceae) named as "catechu" is a plant, the decoction of heartwood of which is daily consumed as thirst quencher by a good percentage of the population in South India. The plant is mainly distributed in India and other Asian countries. It has been used in Indian traditional medicine for the treatment of asthma, bronchitis, colic, diarrhea, boils, skin afflictions, sores and stomatitis. The present investigation was aimed to study the immunomodulatory effects of different fractions of ethanol extract of A. catechu heartwood and HPLC analysis of the active fraction. Three fractions namely, butanol, chloroform and ethyl acetate were prepared from ethanol extract of A. catechu heartwood. Each of these fractions was assessed for its immunomodulatory activity. In vivo immunomodulatory activity was analyzed by sheep red blood cells (SRBC) specific hemagglutinating antibody titer, plaque-forming cell assay and delayed type hypersensitivity (DTH) reaction in Swiss albino mice. In vitro immunomodulating potential of the fractions was studied using murine peritoneal macrophages and splenocytes. Non-specific immune functions such as phagocytosis (nitroblue tetrazolium reduction assay and cellular lysosomal enzyme assay), nitric oxide (NO) production and cytokine release (TNF-α and IL-10) were studied in macrophages. In addition, splenocyte proliferation was also studied. In the in vivo experiments, butanol and chloroform fractions showed an increase in antibody titer dose-dependently. At higher dose (400 mg/kg b. w.) treatment the butanol fraction produced an enhancement in the number of plaque-forming cells (antibody producing cells) in the spleen. SRBC induced DTH reaction was significantly increased with butanol fraction in a dose-dependent manner. Peritoneal macrophages showed an increased phagocytic response on treatment with butanol fraction (100 µg/mL) as evidenced by its effect on nitroblue tetrazolium reduction and cellular lysosomal enzyme activity. All three fractions inhibited the production of NO and the release of TNF-α. Interleukin-10 production was significantly increased after treatment with butanol fraction. High-performance liquid chromatography analysis of the butanol fraction showed the presence of high concentration of catechin. The results suggested that butanol fraction of ethanol extract of A. catechu heartwood had immunomodulatory effects on non-specific, humoral, and cell-mediated immune functions. This study may be useful in validating the rationality of daily consumption of decoction of A. catechu and also its use in traditional medicine system. The study also suggests the possible use of A. catechu in the immunostimulatory herbal preparations.

Protective effect of Rotula aquatica Lour against gentamicin induced oxidative stress and nephrotoxicity in Wistar rats.

Gentamicin is an aminoglycoside antibiotic widely used for the treatment of life-threatening infections caused by Gram-negative bacteria. The use of gentamicin was limited due to its ototoxic and nephrotoxic adverse effects. The current study was designed to evaluate the protective effect of ethyl acetate fraction from Rotula aquatica (EFRA) against gentamicin induced nephrotoxicity. The antioxidant enzymes status, lipid peroxidation, nitrate and ROS level, serum markers like creatinine, Urea, BUN were estimated in the present study. The histopathological analysis of renal tissues was done by H&E and PAS staining. The mRNA level expression of KIM-1, NF-κB, TNF- α, and IL-6 were measured by semi-quantitative reverse transcription-polymerase chain reaction. The changes in antioxidant parameters were restored by the treatment of EFRA at different dose (50 mg/kg bwt, 100 mg/kg bwt). The serum parameters, ROS, MDA and nitrate level were decreased by administration of EFRA. The EFRA ameliorates histological changes associated with gentamicin induced nephrotoxicity. The mRNA level expression of KIM-1, NF-κB, TNF- α, and IL-6 were downregulated in EFRA treated groups. The results from present study reveals the role of EFRA as good anti-inflammatory and nephro protective drug.

Studies on biofilm formation and virulence factors associated with uropathogenic Escherichia coli isolated from patient with acute pyelonephritis.

The current study aims to the detection of pathogenic potential and virulence factor identification of uropathogenic Escherichia coli BRL-17 isolated from patients urine. The organism was isolated from the patient with chronic pyelonephritis. The identification of organism was done by analyzing gram staining, biochemical, 16S rDNA analysis, Raman microscopy and SEM analysis. The pathogenic potential was identified by multiplex PCR analysis of virulence factor genes like sfa, hly D, pap C. The biofilm forming ability was tested by congo red agar assay and tissue culture plate assay. The result of gram staining and biochemical analysis shows the characteristics of E-coli. The 16S rDNA analysis of the clinically isolated uropathogen showed 100% similarity with uropathogenic Escherichia coli strain. Raman microscopy and SEM confirms the organism as E-coli. The Multiplex PCR study identifies virulence genes like sfa, hly D, pap C in isolated E-coli. The presence of P fimbriae coded pap C gene, S fimbriae coded sfa gene and hemolysin-D coded hly D gene discloses its potential to cause urinary tract infection. Biofilm assay result enhances the organism's role as strong biofilm former. This biofilm forming ability of Escherichia coli strain BRL-17 made the organism to escape from host immune system and helps to colonize in bladder and kidney. This also helps to enhance the resistance to antibiotics. Our study confirms the organism as multidrug resistant, highly virulent, strong biofilm forming E-coli. The strain may be used for the development of animal models of pyelonephritis for the purpose of drug discovery.

Antagonism Against Fish Pathogens by Cellular Components/Preparations of Bacillus coagulans (MTCC-9872) and It's In Vitro Probiotic Characterisation.

Bacterial fish pathogens are pervasive in aquaculture. Control of bacterial fish pathogen is a difficult task among aquaculture practitioners. A large number of antibiotics are used for the control of prevalent bacterial pathogens in aquaculture. This may lead to drug resistance among pathogens and further treatment will be ineffective. Here, we can use probiotic bacteria as a biocontrol agent in fish disease and it is a novel field. In this study, antimicrobial potential of the bacterium Bacillus coagulans (MTCC-9872) has been evaluated through in vitro antagonistic activity of cellular preparations/components against potent pathogens. The cellular preparations/components such as Ethyl acetate extract, whole-cell product, heat-killed whole-cell product, and filtered broth were exhibited bactericidal activity against the tested pathogens. Bactericidal activity varied among different cellular preparation/components. The tested bacterium effectively produced biofilm as significant as tested positive control in a microtitre plate and effectively adhered on to the glass slide. In addition, the bacterium was capable of producing extracellular enzymes necessary for the digestion of food materials and was capable to grow in fish mucus from Oreochromis niloticus. The bacterium tolerated bile juice secreted by the host. Moreover, intraperitoneal injection of the bacterium did not induce any pathological signs, symptoms or mortalities in Oreochromis niloticus and revealed the safety of this bacterium in the fish.

Efficient visible light induced synthesis of silver nanoparticles by Penicillium polonicum ARA 10 isolated from Chetomorpha antennina and its antibacterial efficacy against Salmonella enterica serovar Typhimurium.

The green synthesis of silver nanoparticles (AgNPs) using biological systems such as fungi has evolved to become an important area of nanobiotechnology. Herein, we report for the first time the light-induced extracellular synthesis of silver nanoparticles using algicolous endophytic fungus Penicillium polonicum ARA 10, isolated from the marine green alga Chetomorpha antennina. Parametric optimization, including the concentration of AgNO3, fungal biomass, ratio of cell filtrate and AgNO3, pH, reaction time and presence of light, was done for rapid AgNPs production. The obtained silver nanoparticles (AgNPs) were characterized by UV-Visible spectroscopy, Fourier transform infrared (FTIR) spectroscopy, Raman spectroscopy and Transmission electron microscopy (HRTEM-EDAX). The AgNPs showed a characteristic UV-visible peak at 430 nm with an average size of 10-15 nm. The NH stretches in FTIR indicate the presence of protein molecules. The Raman vibrational bands suggest that the molecules responsible for the reduction and stability of AgNPs were extracellular proteins produced by P.polonicum. Antibacterial evaluation of AgNPs against the major foodborne bacterial pathogen Salmonella enterica serovar Typhimurium MTCC 1251, was assessed by well diffusion, Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) assay. Killing kinetic studies revealed complete killing of the bacterial cells within 4 h and the bactericidal nature of synthesized nanoparticles was confirmed by fluorescent microscopy and scanning electron microscopy. Furthermore, the bactericidal studies with Transmission electron microscopy (TEM) at different time intervals explored the presence of AgNPs in the cell wall of S.Typhimurium at about 30 min and the complete bacterial lysis was found at 24 h. The current research opens an insight into the green synthesis of AgNPs and the mechanism of bacterial lysis by direct damage to the cell wall.

Green synthesized silver nanoparticles by marine endophytic fungus Penicillium polonicum and its antibacterial efficacy against biofilm forming, multidrug-resistant Acinetobacter baumanii.

Acinetobacter baumanii, a gram-negative, non-motile, encapsulated coccobacillus which causes infections worldwide. The objective of this study was to find a fungal strain that could be utilized to biosynthesize antibacterial silver nanoparticles (AgNPs) against Acinetobacter baumanii. The present investigation explains rapid and extracellular biosynthesis of silver nanoparticles by the algicolous endophytic fungus, Penicillium polonicum, isolated from the marine green alga Chetomorpha antennina. The obtained silver nanoparticles were characterized by UV-Vis spectroscopy, Raman spectroscopy, Fourier transformation infrared (FTIR), and Transmission electron microscopy (TEM). The SNPs showed a characteristic UV- visible peak at 430 nm with an average size of 10-15 nm. As evident from the FTIR and Raman spectra, possibly the protein components of fungal extract have caused the reduction of silver nitrate. Parametric optimization, including the concentration of AgNO3, ratio of cell filtrate and AgNO3, fungal biomass, reaction time, pH, and presence of light, was done for rapid AgNPs production. The antibacterial efficacy of AgNPs against multi-drug-resistant, biofilm-forming Acinetobacter baumanii, was evaluated by well diffusion assay. The Minimum inhibitory concentration (MIC) of AgNP was 15.62 µgml-1 and the minimum bactericidal concentration (MBC) was 31.24 µgml-1. Killing kinetic assay revealed complete killing of the bacterial cells within 6 h. Log reduction and percent survival of bacterial cells were analyzed from killing kinetic study. Bactericidal nature of synthesized nanoparticles was confirmed by fluorescent microscopical analysis. The effect of AgNPs on the ultrastructure of bacterial pathogen was evaluated by Transmission electron microscopy.

Antibacterial activity of autochthonous bacteria isolated from Anabas testudineus (Bloch, 1792) and it's in vitro probiotic characterization.

Antimicrobial potentials of bacteria isolated from Anabas testudineus have been evaluated through in vitro antagonistic activity against potent fish pathogens. The cellular components and filtered culture medium were effective against six fish pathogens. Altogether 110 strains were isolated from the fish gut, out of which 10 strains were selected through well diffusion method. From them, a strain HGA8B having cumulative maximum score was selected as candidate probiotic. The whole-cell product, heat-killed whole-cell product, Ethyl acetate extract, and the filtered broth were exhibited bactericidal activity against the tested pathogens. In addition, the isolated bacterium was capable of producing extracellular enzymes important for the digestion of food materials and was capable of growth in fish mucus from Oreochromis niloticus. The strain tolerated bile juice secreted by the host and effectively produced biofilm. Analysis of 16S rDNA sequence revealed that isolated strain HGA8B was Bacillus sp. (MF351637). Furthermore, intraperitoneal injection of the bacterium did not induce any pathological signs, symptoms or mortalities in Oreochromis niloticus and revealed the safety of this bacterium as a candidate probiotic in aquaculture.

Antibacterial activity and probiotic characterization of autochthonous Paenibacillus polymyxa isolated from Anabas testudineus (Bloch, 1792).

Microbial fish pathogens are prevalent in aquaculture. Control of bacterial fish pathogens is important and bio control of pathogenic bacteria is a novel field of study. The aim of this study was to evaluate the antagonistic activity of bacteria isolated from Anabas testudineus against potent fish pathogens. The cellular components/preparations and filtered cell free culture supernatants were effective against six fish pathogens. Altogether 110 strains were isolated from fish proximal and distal intestine, out of which 10 strains were selected through well diffusion method. From them a strain HGA4C having prominent antimicrobial activity was selected as candidate probiotic strain. The whole-cell product, heat-killed whole-cell product and the filtered broth were exhibited bactericidal activity against the tested pathogens. Among them cell free culture supernatant showed maximum inhibition. In addition, isolated candidate probiotic bacterium was capable of producing extracellular enzymes important for the digestion of food ingredients and was effectively grown in fish mucus obtained from Oreochromis niloticus. The strain tolerated gradient of bile juice secreted by the host and effectively produced biofilm. Analysis of 16S rDNA sequence revealed that isolated strain HGA4C was Paenibacillus polymyxa (MF457398.1). Furthermore intraperitoneal injection of the bacterium did not induce any pathological anomalies or mortalities in Oreochromis niloticus and disclosed the safety of this bacterium as a candidate probiotic in aquaculture.

The influence of tissue antioxidant enzymes and inflammatory cascade in pathology of cystitis: An experimental rat model.

We evaluate the role of antioxidant enzyme status and inflammatory cascade in disease progression of cystitis in a rat model. The animals were injected with clinically isolated Uropathogenic Escherichia coli (UPEC) and study the effect of various antioxidant enzymes and inflammatory markers in disease pathology on the 0th day, 12 h and 7th day of infection. The antioxidant status of bladder tissue was decreased during the 7th day of infection. Lipid peroxidation marker MDA was increased on the 7th day of infection in rats. The histopathology of bladder tissue shows severe inflammation and edema. This study reveals the role of decreased antioxidant status during infection play a vital role in upregulation of inflammation and tissue destruction.

Bacopaside N1 biosynthetic potential of endophytic Aspergillus sp. BmF 16 isolated from Bacopa monnieri.

Chemically unique environment of endophytes makes them to have various adaptive mechanisms for survival. One of such mechanisms involves the production of pharmacologically significant plant-specific metabolites. In the present study, 26 endophytic fungi were isolated from stem of Bacopa monnieri (L.) Wettst. plants. All the isolates were screened for bacopaside production property by HPLC. Among these, the fungal isolate BmF 16 which was identified as Aspergillus sp. was confirmed for bacopaside N1 production (m/z 796) by LC-MS/MS analysis. As the extract of BMF16 used in the study was prepared from the fifth generation of culture, the obtained result can be confirmed as due to fungal production of bacopaside. In addition, this property was identified only for one among the 26 fungi screened. As bacopaside N1 production in fungi has not yet been reported, the results of the study are novel.

Role of antioxidant defence, renal toxicity markers and inflammatory cascade in disease progression of acute pyelonephritis in experimental rat model.

Urinary tract infections are the most common bacterial infections affecting millions of people each year worldwide. The animal model provides an excellent and suitable system for studying cystitis and pyelonephritis caused by Escherichia coli and other uropathogens. Using this established model, we evaluate the role of antioxidant defence system, renal injury markers, and blood parameters in the diseases progression during Escherichia coli infection on 0th day, 12h and 7th day. The antioxidant enzymes like SOD, CAT, GSH, GPx, GR levels were evaluated. The blood parameters like AST, ALT, ALP, Total protein, BUN, creatinine level were estimated in infection model. The relative organ weights, anti microbial status of kidney, CRP, WBC count were done for the evaluation of inflammatory response associated with the infection. The oxidative stress marker like MDA was also evaluated. Histopathological analysis of renal tissue provides direct vision to tissue damage. The antioxidant status of renal tissue was decreased during the 7th day of infection. Likewise, renal toxicity markers were significantly increased during bacterial infection. The inflammatory markers like CRP, WBC count and oxidative stress marker like MDA were significantly increased by the infection on 7th day. The histopathology of renal tissue also reveals the inflammation and tissue damage associated with acute pyelonephritis.