The emergence of multidrug-resistant Proteus vulgaris infection in cage reared Pangasianodon hypophthalmus: Molecular characterization and host-pathogen response.

The study investigates the causative agent responsible for massive mortality in Pangasianodon hypophthalmus cage farms. The infected pangasius were lethargic, not taking feed, and had exophthalmia, deep ulceration, and hemorrhage on the ventral body surface. Pathogens were isolated from infected pangasius tissue samples, and the strain was preliminarily identified as Proteus vulgaris based on morphology, biochemical tests, 16S rRNA PCR sequencing, and phylogenetic analysis. The microbiological analysis revealed that P. vulgaris was associated with the disease outbreak and mortality of P. hypophthalmus, as confirmed by the biofilm formation, swimming activity, and survival assay results. The isolated P. vulgaris was resistant to several antibiotics, including Doxycycline, Dicloxacillin, Polymyxin B, Chloramphenicol, and Imipenem, exhibiting multiple antibiotic-resistant phenotypes. Furthermore, a host-pathogen model was developed to investigate the in vivo effect of emerging P. vulgaris infection in striped catfish. Results showed that P. vulgaris infection exhibited varying degrees of cellular changes in the kidney, liver, and gill tissues of infected P. hypophthalmus samples. The transcription analysis further highlights that P. vulgaris modulates the antioxidants, immune activation, growth regulator, homeostasis, degradation of invading DNA, and DNA damage-related gene expression in liver, kidney, spleen, skin, and gill tissue samples of infected P. hypophthalmus. The lessons learned from the study could be critical in understanding the P. vulgaris infection patterns and pathobiology necessary to enable risk assessment and develop management measures to control the bacterium's virulence.

Deciphering the impact of endoparasitic infection on immune response and gut microbial composition of Channa punctata.

Intestinal parasitic infections caused by helminths are globally distributed and are a major cause of morbidity worldwide. Parasites may modulate the virulence, gut microbiota diversity and host responses during infection. Despite numerous works, little is known about the complex interaction between parasites and the gut microbiota. In the present study, the complex interplay between parasites and the gut microbiota was investigated. A total of 12 bacterial strains across four major families, including Enterobacteriaceae, Morganellaceae, Flavobacteriaceae, and Pseudomonadaceae, were isolated from Channa punctata, infected with the nematode species Aporcella sp., Axonchium sp., Tylencholaimus mirabilis, and Dioctophyme renale. The findings revealed that nematode infection shaped the fish gut bacterial microbiota and significantly affected their virulence levels. Nematode-infected fish bacterial isolates are more likely to be pathogenic, with elevated hemolytic activity and biofilm formation, causing high fish mortality. In contrast, isolates recovered further from non-parasitised C. punctata were observed to be non-pathogenic and had negligible hemolytic activity and biofilm formation. Antibiogram analysis of the bacterial isolates revealed a disproportionately high percentage of bacteria that were either marginally or multidrug resistant, suggesting that parasitic infection-induced stress modulates the gut microenvironment and enables colonization by antibiotic-resistant strains. This isolation-based study provides an avenue to unravel the influence of parasitic infection on gut bacterial characteristics, which is valuable for understanding the infection mechanism and designing further studies aimed at optimizing treatment strategies. In addition, the cultured isolates can supplement future gut microbiome studies by providing wet lab specimens to compare (meta)genomic information discovered within the gut microenvironment of fish.

Forecast of agri-residues generation from rice, wheat and oilseed crops in India using machine learning techniques: Exploring strategies for sustainable smart management.

Management of agri-residues generated in large quantities necessitates for its accurate estimation. Data analysis using machine learning methods can predict the agri-residues generation. The objective of the study was to forecast agri-residues generation from rice, wheat, and oilseed crops in India using ML methods and their sustainable uses. Prediction of agri-residues was done first by forecasting the crop production via the application of ML techniques for the period 2022 to 2030, and then the amount of crop residues generation calculated by multiplying the crop productions with the residues-to-product-ratio (RPR) values of the respective crops. RPR was estimated by using the gravimetric ratio of the residue to the actual crop production. The crop-specific RPR values were taken from various earlier studies in Indian context. The RPR values of 1.73 for the rice, 1.65 for wheat, and 2.6 for the oilseed crop were used as a conversion factor for residues calculation. Machine learning models linear regression, sequential minimal optimization regression (SMOreg), M5 Rule, and Gaussian process were used in the study. SMOreg performed better in models tested by coefficient of determination, root mean square error, and mean absolute error. The models predicted the generation of residues in 2030 as rice straw and husk 195.76 Mt to 277.68 Mt, wheat straw 188.62 Mt to 266.95 Mt, and oilseed stalk and oil cakes 55.61 Mt to 96.30 Mt in India. An overview of the management of agri-residues discussed. Estimation of agri-residues can provide an opportunity to utilize them with the best possible ways, lessen pollution and promote a zero-waste strategy.

Agri-residues and agro-industrial waste substrates bioconversion by fungal cultures to biocatalyst lipase for green chemistry: A review.

Huge amounts of agri-residues generated from food crops and processing are discarded in landfills, causing environmental problems. There is an urgent need to manage them with a green technological approach. Agri-residues are rich in nutrients such as proteins, lipids, sugars, minerals etc., and provide an opportunity for bioconversion into value-added products. Considering the importance of lipase as a biocatalyst for various industrial applications and its growing need for economic production, a detailed review of bioconversion of agri-residues and agro-industrial substrate for the production of lipase from fungal species from a technological perspective has been reported for the first time. Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) flow diagram was used for the identification and selection of articles from ScienceDirect, Google Scholar, and Scopus databases from 2010 to 2023 (July), and 108 peer-reviewed journal articles were included based on the scope of the study. The composition of agri-residues/agro-industrial wastes, fungal species, lipase production, industrial/green chemistry applications, and the economic impact of using agri-residues on lipase costs have been discussed. Bioconversion procedure, process developments, and technology gaps required to be addressed before commercialization have also been discussed. This process expects to decrease the environmental pollution from wastes, and low-cost lipase can help in the growth of the bioeconomy.

Sc(OTf)3-Mediated One-Pot Synthesis of Coumarin-Fused Furans: A Thiol-Dependent Reaction for the Easy Access of 2-Phenyl-4H-furo[3,2-c]chromen-4-ones.

Herein, we report a simple and efficient method for the preparation of novel thioether-linked coumarin-fused furans from the one-pot three-component reaction of arylglyoxal, 4-hydroxycoumarin, and various aromatic thiols in the presence of Sc(OTf)3 as a catalyst. This methodology is also applicable to cyclic 1,3-dicarbonyls such as cyclohexane-1,3-dione and dimedone. Depending upon the thiols, this methodology can either give a three-component thioether-linked coumarin-fused furan (4) or a two-component furocoumarin product (5). Wide substrate scope, good to excellent yields, and products having more than one pharmaceutically important motif are the salient features of this methodology.

Photofermentative biohydrogen generation from organic acids by Rhodobacter sphaeroides O.U.001: Computational fluid dynamics modeling of hydrodynamics and temperature.

Hydrogen gas is a clean-burning fuel suitable for powering public vehicles. Hydrogen fuel has the highest energy density (143 MJ kg-1 ). This research paper emphasizes three-dimensional hydrodynamics and temperature distribution during photobiohydrogen generation by Rhodobacter sphaeroides strain O.U.001 in a triple-jacketed 1 L photobioreactor (PBR). The fermentation broth has turbulent flow conditions and light gradients among various layers, which affect the light conversion efficiency of the PBR. From the carbon source (malic acid), various organic acids are produced within fermentation (lactate, acetate, and formate). Modeling and simulation studies by computational fluid dynamics confirmed uniform fluid dynamics and heat transfer throughout the annular PBR. The modified Gompertz equation gave good simulated fitting with an experimental value for H2 generation. R. sphaeroides O.U. 001 gave good simulated results for H2 generation with mathematical modeling of substrate consumption kinetics and substrate utilization for biomass.

Improvement in Production of Rhamnolipids Using Fried Oil with Hydrophilic Co-substrate by Indigenous Pseudomonas aeruginosa NJ2 and Characterizations.

Commercialization of biosurfactant remained a challenge due to lack of structural variation and economical process using low-cost materials and low productivity. Improvement in production of biosurfactant using fried oil with hydrophilic co-substrate by an indigenous strain was studied. Microbe isolated from exhaust chimney condensate was screened for utilization of mixed carbon source and then identified as Pseudomonas aeruginosa NJ2 by 16S rDNA gene sequence. FTIR, HPLC, and NMR analyses confirmed that biosurfactant was rhamnolipids. Batch fermentation using mixed substrates improved the cell growth yield to 1.48 g/L (2.34 times) and product yield to 4.28 g/L (3.4 times) with maximum specific growth rate 0.1 h-1 (two times) and specific production rate 0.5 h-1 (13 times) due to higher cell density and direct synthesis of lipid and rhamnose moieties through central metabolic pathways of the two substrates. Increase in carrying capacity and coefficient value (two times) of logistic equation confirmed the significance of mixed substrates. The biosurfactant showed excellent surface active and thermo-chemical stability properties. Economical production of biosurfactant with high yield and productivity could be possible by isolation of mixed carbon source utilizing strain and optimization of waste substrates from oil/soapstock and sugar/corn syrup industries in media.

I2/DMSO mediated multicomponent reaction for the synthesis of 2-arylbenzo[d]imidazo[2,1-b] thiazole derivatives.

Synthesis of a series of 2-arylbenzo[d]imidazo[2,1-b] thiazoles tethered with barbituric acid moiety has been reported from the three component reaction of 2-aminobenzothiazoles, barbituric acids and terminal aryl acetylenes or aryl methyl ketones in the presence of I2 in DMSO medium. Both conventional and microwave heating conditions can be used for this multicomponent reaction. The salient features of this methodology are: (i) formation of one C-C and two C-N bonds in one-pot under metal-free oxidation followed by cyclization, (ii) selective formation of the fused imidazole ring, (iii) wide substrate scope, (iv) easy purification of the products, (v) products having more than one pharmaceutically important motifs and (vi) gram scale synthesis possible.

The complete mitochondrial genome sequence of Osteobrama belangeri (Cyprinidae) and its comparison with other related Cypriniformes fish species.

The complete mitogenome of Osteobrama belangeri is described using Ion Torrent (PGM sequencer), which was 16,609 bp in size comprising 13 mRNAs, teo rRNA genes, 22 tRNAs, and 926 bp as D-Loop control region, in addition to gene order and organization, being similar to most of the other related Cypriniformes fish mitogenome of NCBI databases. The all 22 tRNAs were packed into a typical clover-leaf structure. In the present study, the mitogenome has 99% similarity to the complete mitogenome sequence of O. belangeri mitogenome details previously and also would be helpful in understanding the phylogenetics, population genetics, and evolution of family Cyprinidae fishes.

Production of laccase by repeated batch semi-solid fermentation using wheat straw as substrate and support for fungal growth.

Repeated batch semi-solid fermentation (sSF) process using wheat straw substrate and fungal growth of Ganoderma lucidum on solid substrate was studied for production of laccase. pH showed significant effect on laccase production. Highest laccase activity with pH controlled to 5.0 in batch sSF was 15257.2 ± 353.4 U L- 1 on 9th day. In repeated batch process at pH 5.0, insoluble biomass substrate and fungal growth were reused after liquid part of medium was replaced with glucose, ammonium phosphate (best nitrogen source) and combined glucose and ammonium phosphate solution separately. Refilled to 80% w v- 1 of initial soluble sugar of first batch resulted in highest laccase production with peak activity after 4 days from replacement. Production of enzyme increased from 15257.2 U L- 1 in first batch to cumulative 90164.4 U L- 1 in 29 days after six repeated batches, productivity increased from 1680.2 to 3110.3 U L- 1 day- 1 (∼ 1.9 times) due to reductions in inhibitory effects and time required for fungal growth. Utilization of wheat straw in repeated batch sSF was supported by composition analysis and morphological changes (scanning electron microscopy) of substrate. Economic production of laccase using agricultural residues in repeated batch sSF could be possible.

Synthesis of fused pyrroles containing 4-hydroxycoumarins by regioselective metal-free multicomponent reactions.

The reaction of arylglyoxals, 4-hydroxycoumarin, and aromatic amines such as 7-amino-2-methylchromone, 6/7-aminoflavone, 7-amino-4-methylcoumarin, 1-amino-9-fluorenone, 1-aminoanthraquinone and aniline derivatives in acetic acid medium under microwave conditions provides the corresponding regioselective fused pyrroles having hydroxycoumarin and aryl substituents. Alternatively, we have developed another method using in situ arylglyoxals from acetophenone derivatives by I2/DMSO promoted C-H oxidation followed by one-pot three component cyclization reactions to provide similar fused pyrroles. Using both the methods a series of novel pyrroles fused with pharmacologically important chromone, flavone, coumarin, fluorenone, and anthraquinone moieties were synthesized under metal-free reaction conditions in good to very good yields within a short reaction time. The structures of the synthesized fused pyrroles have been unambiguously confirmed by spectroscopic techniques, mass analysis and single crystal XRD.

Synthesis of Pyrimidine Fused Quinolines by Ligand-Free Copper-Catalyzed Domino Reactions.

Herein, we report two novel methods for the synthesis of pyrimidine fused quinolines using a one-pot C-C and C-N bond forming strategy from the reaction of 6-aminouracils with 2-bromobenzaldehydes or 2-bromobenzyl bromide derivatives in the presence of 10 mol % CuCl2 without using any ligand. The reaction of 2-bromobenzaldehyde or its derivatives with 6-aminouracils in the presence of K2CO3 as base and a catalytic amount of CuCl2 in DMF medium under microwave heating conditions provides corresponding pyrimidine fused quinoline derivatives in good yields within 30 min. Alternatively, pyrimidine fused quinoline derivatives have been synthesized from the reaction of 2-bromobenzyl bromides with 6-aminouracil derivatives in the presence of molecular oxygen, CuCl2 (10 mol %), and K2CO3 as base in DMF under reflux conditions. Structures of all the products were unambiguously confirmed by spectroscopic techniques and by recording single crystal XRD of 3a.

Effects of wheat straw solid contents in fermentation media on utilization of soluble/insoluble nutrient, fungal growth and laccase production.

The objective of the work was to study the effect of agri-residue solid contents (2-20% w v-1) in fermentation medium on fungal growth, soluble and insoluble nutrient consumption and laccase production. Fungal strain Ganoderma lucidium and wheat straw substrate was screened for maximum laccase production. At low solid content submerged fermentation (SmF), fungus utilized mainly soluble nutrient and was unable to access the insoluble nutrient in media due to lack of contact with solid. At high solid content solid-state fermentation (SF), fungi grew on solid surface with dense and thin hyphae, utilized mainly insoluble nutrient. At medium solid content (8% w v-1) semi-solid fermentation (sSF), fungi grew on solid substrates with network of thick intercrossed hyphae, utilized both soluble and insoluble nutrients optimally resulting in highest fungal growth and laccase activity (~ 3.5 folds than in SmF and ~ 2.5 folds than in SF). Importance of soluble and insoluble nutrients was also established after isolation of their individual effects. Morphology of fungal growth (SEM), composition, thermal analysis (TGA/DTG) of substrates confirmed the results. sSF showed potential for the production of enzymes through utilization of agricultural residues as substrate.

Fungal pretreatment of sweet sorghum bagasse with supplements: improvement in lignin degradation, selectivity and enzymatic saccharification.

Sweet sorghum bagasse (SSB) from food processing and agricultural industry has attracted the attention for uses in production of biofuel, enzymes and other products. The alteration in lignocellulolytic enzymes by use of supplements in fungal pretreatment of SSB to achieve higher lignin degradation, selectivity value and enzymatic hydrolysis to fermentable sugar was studied. Fungal strain Coriolus versicolor was selected for pretreatment due to high ligninolytic and low cellulolytic enzyme production resulting in high lignin degradation and selectivity value. SSB was pretreated with supplements of veratryl alcohol, syringic acid, catechol, gallic acid, vanillin, guaiacol, CuSO4 and MnSO4. The best results were obtained with CuSO4, gallic acid and syringic acid supplements. CuSO4 increased the activities of laccase (4.9-fold) and polyphenol oxidase (1.9-fold); gallic acid increased laccase (3.5-fold) and manganese peroxidase (2.5-fold); and syringic acid increased laccase (5.6-fold), lignin peroxidase (13-fold) and arylalcohol oxidase (2.8-fold) resulting in enhanced lignin degradations and selectivity values than the control. Reduced cellulolytic enzyme activities resulted in high cellulose recovery. Enzymatic hydrolysis of pretreated SSB yielded higher sugar due to degradation of lignin and reduced the crystallinity of cellulose. The study showed that supplements could be used to improve the pretreatment process. The results were confirmed by scanning electron microscopy, X-ray diffraction, Fourier transform infrared spectroscopy and thermogravimetric/differential thermogravimetric analysis of SSB.

Enhancement in multiple lignolytic enzymes production for optimized lignin degradation and selectivity in fungal pretreatment of sweet sorghum bagasse.

The objective of this work was to study the increase in multiple lignolytic enzyme productions through the use of supplements in combination in pretreatment of sweet sorghum bagasse (SSB) by Coriolus versicolor such that enzymes act synergistically to maximize the lignin degradation and selectivity. Enzyme activities were enhanced by metallic salts and phenolic compound supplements in SSF. Supplement of syringic acid increased the activities of LiP, AAO and laccase; gallic acid increased MnP; CuSO4 increased laccase and PPO to improve the lignin degradations and selectivity individually, higher than control. Combination of supplements optimized by RSM increased the production of laccase, LiP, MnP, PPO and AAO by 17.2, 45.5, 3.5, 2.4 and 3.6 folds respectively for synergistic action leading to highest lignin degradation (2.3 folds) and selectivity (7.1 folds). Enzymatic hydrolysis of pretreated SSB yielded ∼2.43 times fermentable sugar. This technique could be widely applied for pretreatment and enzyme productions.

Improvement of selective lignin degradation in fungal pretreatment of sweet sorghum bagasse using synergistic CuSO4-syringic acid supplements.

Sweet sorghum bagasse (SSB) generated in large quantities could be hydrolyzed to sugar and then fermented to green fuels. The hydrolysis of SSB polysaccharides interlocked in recalcitrant lignin network is the major problem. Pretreatment of SSB in SSF by using Coriolus versicolor with CuSO4-syringic acid supplements for effects on production of ligninocellulolytic enzymes, lignin degradation and selectivity values (SV) were studied. C. versicolor was selected based on high ligninolytic and low cellulolytic abilily. Individually, CuSO4 increased the activities of laccase (4.9 folds) and PPO (1.9 folds); syringic acid increased LiP (13 folds), AAO (2.8 folds) and laccase (5.6 folds) resulting in increased lignin degradation and SVs. Combined syringic acid (4.4 µmol g-1 SSB) and CuSO4 (4.4 µmol g-1 SSB) increased the activities of laccase, LiP, MnP, PPO and AAO by 11.2, 17.6, 2.8, 2.4 and 2.3 folds respectively due to synergistic effect, resulting in maximum lignin degradation 35.9 ± 1.3% (w w-1) (1.86 fold) and highest SV 3.07 (4.7 fold). Enzymatic hydrolysis of pretreated SSB yielded higher (∼2.2 times) fermentable sugar. Pretreated SSB was characterized by XRD, SEM, FTIR and TGA/DTG analysis to confirm results. It is possible to improve fungal pretreatment of agricultural waste by combination of supplements.

Fungal Pretreatment of Sweet Sorghum Bagasse with Combined CuSO4-Gallic Acid Supplement for Improvement in Lignin Degradation, Selectivity, and Enzymatic Saccharification.

Sweet sorghum (Sorghum sp.) has high biomass yield. Hydrolysis of lignocellulosic sweet sorghum bagasse (SSB) to fermentable sugar could be useful for manufacture of biofuel or other fermentation products. Pretreatment of lignocellulosic biomass to degrade lignin before enzymatic hydrolysis is a key step. Fungal pretreatment of SSB with combined CuSO4-gallic acid supplements in solid-state fermentation (SSF) to achieve higher lignin degradation, selectivity value (SV), and enzymatic hydrolysis to sugar was studied. Coriolus versicolor was selected due to high activities of ligninolytic enzymes laccase, lignin peroxidase (LiP), manganese peroxidase (MnP), polyphenol oxidase (PPO), and arylalcohol oxidase (AAO) and low activities of cellulolytic enzymes CMCase, FPase, and ß-glucosidase with high lignin degradation and SV in 20 days. CuSO4/gallic acid increased the activities of ligninolytic enzymes resulting in enhanced lignin degradations and SVs. Cumulative/synergistic effect of combined supplements further increased the activities of laccase, LiP, MnP, PPO, and AAO by 7.6, 14.6, 2.67, 2.06, and 2.15-folds, respectively (than control), resulting in highest lignin degradation 31.1 ± 1.4% w/w (1.56-fold) and SV 2.33 (3.58-fold). Enzymatic hydrolysis of pretreated SSB yielded higher (~2.2 times) fermentable sugar. The study showed combined supplements can improve fungal pretreatment of lignocellulosic biomass. XRD, SEM, FTIR, and TGA/DTG of SSB confirmed the results.

Cotton Stalk Pretreatment Using Daedalea flavida, Phlebia radiata, and Flavodon flavus: Lignin Degradation, Cellulose Recovery, and Enzymatic Saccharification.

Lignocellulolytic enzyme activities of selective fungi Daedalea flavida MTCC 145 (DF-2), Phlebia radiata MTCC 2791 (PR), and non-selective fungus Flavodon flavus MTCC 168 (FF) were studied for pretreatment of cotton stalks. Simultaneous productions of high LiP and laccase activities by DF-2 during early phase of growth were effective for lignin degradation 27.83 ± 1.25 % (w/w of lignin) in 20-day pretreatment. Production of high MnP activity without laccase in the early growth phase of PR was ineffective and delayed lignin degradation 24.93 ± 1.53 % in 25 days due to laccase production at later phase. With no LiP activity, low activities of MnP and laccase by FF yielded poor lignin degradation 15.09 ± 0.6 % in 20 days. Xylanase was predominant cellulolytic enzyme produced by DF-2, resulting hemicellulose as main carbon and energy source with 83 % of cellulose recovery after 40 days of pretreatment. The glucose yield improved more than two fold from 20-day DF-2 pretreated cotton stalks after enzymatic saccharification.

Synthesis of Novel 5,6-Disubstituted Pyrrolo [2,3-d]Pyrimidine-2,4-Diones Via One-Pot Three-Component Reactions.

A simple and novel method for the synthesis of 5,6-disubstituted pyrrolo[2,3-d]pyrimidine-2,4-diones has been reported using arylglyoxal-based three-component reactions. Under microwave heating conditions, arylglyoxal, 6-amino uracil, or its derivatives reacts with various thiols in acetic acid medium to provide a series of pyrrolo[2,3-d]pyrimidine-2,4-diones (4) having a thioether and an aryl ring in 5 and 6 positions, respectively. On the other hand reaction of arylglyoxal, amino uracil and malononitrile in place of thiols, provided corresponding 5,6-disubstituted pyrrolo[2,3-d]pyrimidine-2,4-diones (5) with selectively converting one of the -CN to -CONH2 group both in conventional (method A) and microwave heating conditions (method B). This methodology is a simple and efficient protocol for the synthesis of diverse 5,6-disubstituted pyrrolo[2,3-d]pyrimidine-2,4-diones from the readily available starting materials in good yields.