Identification of Bacterial Isolates and Their Antimicrobial Susceptibility Pattern from Wound/Pus Sample in a Tertiary Care Hospital, Gwalior, India

Objective: The goal of this investigation was to look at the frequency and dispersal of bacteria isolated from pus/wound, as well as their susceptibility patterns. Materials and Methods?A study was conducted on 175 patients who provided pus and/or wound discharge samples in different wards (outpatient department or inpatient department). MacConkey agar and blood agar plates were immediately inoculated with samples and incubated at 37°C for 24?hours. The Gram stain and biochemical tests were used to identify all isolates after incubation. Kirby–Bauer's disc diffusion method was used to perform sensitivity tests on Mueller–Hinton agar plates. Results?This study covered 175 patients, with a bacterial isolation rate of 102 (58.28%). Males outnumbered females in the samples (M:F?=?1.8:1), with a median age of 45 years as majority were in the age group of 40 to 60 years which was 41 (40.20%). Total 90.1% samples showed monomicrobial infection, whereas 9.8% showed polymicrobial infection, and total 112 bacterial strains were isolated. Conclusion?Escherichia coli was the most prevalent isolate in present investigation, followed by Pseudomonas aeruginosa. Chloramphenicol is the only antibiotic which is effective for both gram-negative bacilli and gram-positive cocci. This report's susceptibility statistic may be worth considering for developing empiric treatment regimens for pyogenic infections.

Evaluation of the heavy metals tolerant UV rays treated bacteria isolated from anthropogenic sites of Chambal region, India

Industrial waste is released into the environment and leads to various types of heavy metal, which are toxic, mutagenic and carcinogenic in nature. Heavy metals are not biodegradable but accumulated by living organisms and cause diseases at even low concentrations. In this study, we selected four anthropogenic sites from Chambal region, isolated bacteria and investigated its heavy metal removal capability. The bacteria was isolated and identified as Escherichia coli (Ag-5), on the basis of biochemical and 16S rRNA gene sequence. Among the five (cadmium, cobalt, lead, nickel and zinc) heavy metals studied, Ni2+ has been observed to be highly toxic with minimum inhibitory concentration score of 200 ppm. E. coli could tolerate Zn2+ (300 ppm), Cd2+ (400 ppm), Co2+ (400 ppm) and Pb+2 (500 ppm). Heavy metal tolerance capability was also evaluated by UV rays treated E. coli (Ag-5) isolate and compared with wild strain Ag-5. The result indicated that the tolerance capability was enhanced by UV rays treated bacterial isolate as compared to wild strain with respect to all tested heavy metals. Atomic absorption spectroscopy results revealed that wild strain removed 78.2% cadmium nitrate, while UV rays 30 and 60 s. exposed strain removed 85.9 and 83% cadmium nitrate. Wild strain removed 64.4% nickel chloride, while UV rays 30 and 60 s exposed strain removed 66.9 and 74.5% nickel chloride. The results indicate that indigenous E. coli treated with UV rays could serve as heavy metal tolerant bacteria and utilized in bioremediation processes.

Optimization of simultaneous saccharification and fermentation parameters for sustainable production of ethanol from wheat straw by Pichia stipitis NCIM 3498

Bioethanol is a potentially safe and renewable alternate source of energy. However, ethanol production from value added food and feedstock has not shown growth as estimated. Of late, the second generation processes of production of ethanol, such as from lignocellulosic biomass out of agricultural/domestic waste, has been gaining considerable momentum. Here, we attempted optimizing the conditions of physiochemical pretreatment as well as fermentation process using wheat straw by Pichia stipitis NCIM 3498 (now known as Schefferomyces stipitis). We have also studied the influence of process variables, such as incubation temperature, inoculum concentration and different nutrients, on ethanol production. Pulverized wheat straw consists of 32±0.31% cellulose, 48±0.37% hemicellulose and 17±0.15% lignin on dry solid (DS) basis. Wheat straw delignified with 1% HNO3, yielded 11.54% xylose and 1.54% glucose under steam explosion [15 psi (121°C) for 60 min], with a hydrolytic efficiency of 59.56%. Simultaneous Saccharification and Fermentation (SSF) of pretreated wheat straw by crude cellulose (produced by Trichoderma reesei NCIM 1052) and S. stipitis were investigated in the present study. Important process variables for ethanol production from pretreated wheat straw were optimized using response surface methodology (RSM) based on central composite design (CCD) experiments. A three level CCD experiments with central and axial points was used to develop a statistical model for optimization of process variables viz. incubation temperature (30, 32 and 34°C) X1, inoculum level (2, 4 and 6%) X2 and nutrients (1/2/3) X3. Data obtained from RSM on ethanol production were subjected to the analysis of variance (ANOVA), analyzed using a second order polynomial equation, and contour plots were used to study the interactions among three relevant variables of the fermentation process. The fermentation experiments were carried out at flask level. The processing parameters setup for reaching a maximum response for ethanol production was obtained when applying the optimum values for temperature (34°C), inoculum level (6%) and fermentation medium (ammonium sulphate, KH2PO4, peptone and yeast extract) for S. stipitis. Maximum ethanol concentration 7.15 g/L was obtained after 72 h from S. stipitis at the optimized process conditions in anaerobic batch fermentation.

Biovalorization potential of peels of Ananas cosmosus (L.) Merr. for ethanol production by Pichia stipitis NCIM 3498 & Pachysolen tannophilus MTCC 1077.

Bioethanol, is a potential alternate source of energy, renewable and safe. Ethanol production from value added food and feedstock has also not shown growth as estimated. Of late, the second generation processes of production of ethanol, such as from lignocellulosic biomass out of agricultural/domestic waste has been gaining considerable momentum. Here, we explored a new approach for optimizing the conditions of physiochemical pretreatment as well as fermentation process using peels of Ananas cosmosus as substrate and immobilized yeast Pachysolen tannophilus MTCC 1077 and Pichia stipitis NCIM 3498. We have also studied the influence of process variables such as incubation temperature, inoculum concentration and different nutrients on ethanol production. Pulverized peels of A. cosmosus recorded 25 ± 0.31% cellulose, 28 ± 0.18% hemicellulose and 8 ± 0.07% of lignin on dry solid (DS) basis. Peels of A. cosmosus delignified with 1% H2SO4 yielded 18.89% glucose, 38.81% xylose and 29.31% fructose under thermochemical pretreatment using autoclave (121°C, 20 min.), with a hydrolytic efficiency of 75.52 ± 0.45%. FTIR spectroscopy results not only indicated the penetration of H2SO4 in the amorphous region of the biomass and degradation of hemicelluloses but also showed the structural differences before and after pretreatment. The enzymes required for hydrolysis were prepared from culture supernatants of Trichoderma reesei NCIM 1052 using wheat bran as carbon source under submerged fermentation conditions on rotatory shaker incubator (at 28°C for 10 days ). Enzyme activity (U/ml) of crude cellulase produced by T. reesei NCIM 1052 was 311.1 µmole/ml/min. Delignified A. cosmosus peel yielded 51.71 ± 0.44 g/l glucose when enzymatically hydrolysed by crude cellulase at the substrate enzyme ratio of 1:5. Simultaneous saccharification and fermentation (SSF) of peels of A. cosmosus by crude cellulase and separately entrapped Pichia stipitis NCIM 3498 (now known as Scheffersomyces stipitis) and Pachysolen tannophilus MTCC 1077 cells in calcium alginate beads were also investigated in the present study. The fermentation experiments were carried out at flask level. The processing parameters setup for reaching a maximum response for ethanol production was obtained when applying the optimum values for temperature (32°C), inoculum level (6%) and fermentation medium (ammonium sulphate, KH2PO4, peptone and yeast extract) for P. tannophilus MTCC 1077 and temperature (30°C), inoculum level (2%) and fermentation medium (ammonium sulphate, KH2PO4, peptone and yeast extract) for S. stipitis NCIM 3498. Maximum ethanol concentration 10.5 g/l and 10.9 g/l was obtained from P. tannophilus MTCC 1077 and S. stipitis NCIM 3498, respectively at the optimized process conditions in anaerobic batch fermentation.

Analysis of time-dependent recovery from beryllium toxicity following chelation therapy and antioxidant supplementation.

Efforts have been made to minimize the toxic effect caused by beryllium. Adult cyclic rats of Sprague Dawley strain were administered a bolus dose of 50mg/kg beryllium nitrate intramuscularly. The chelation therapy with glutathione (GSH), dimercapto propane sulfonic acid (DMPS)+ selenium (Se) and D-Penicillamine (DPA) + Se was given for 3 days followed by a rest of 1,3 and 7 days respectively. The results revealed a significant fall in the blood sugar level, serum alkaline phosphatase activity, serum proteins. A significant rise in the transaminases i.e. aspartate aminotranferase and alanine aminotranferase pattern is indicative of leakage of enzymes from liver resulting in alterations in the cell permeability. A rise in the hepatic lipid peroxidation activity is a direct indication of oxidative damage resulting in free radical generation. Results of the distribution studies by atomic absorption spectrophotometry reveal an increased concentration of beryllium in liver and kidney followed by lung and uterus. The relative ability of 3 chelating agents to act as antagonists for acute beryllium poisoning have been examined in liver, kidney, lungs and uterus. The appreciable change in the beryllium concentration in various organs is duration-dependent during the entire period being highly significant after 7 days rest. From the biochemical assays, and distribution studies it can be assumed that DPA+Se was the most effective therapeutic agent followed by DMPS+Se and GSH. Thus it can be concluded that DPA+Se is a better therapeutic agent as compared to DMPS+Se and GSH.

Role of chelating agents and antioxidants in beryllium induced toxicity.

The present study was conducted to evaluate the therapeutic effectiveness of chelating agents [glutathione, 2,3 dimercapto propane sulfonic acid (DMPS) and D-penicillamine (DPA)] in combination with antioxidant (sodium selenite) in beryllium induced toxicity in female rats. A bolus dose of 50mg/kg-beryllium nitrate was administered singly followed by chelation therapy with GSH, DMPS + Se and DPA + Se at various durations of 1,3 and 7 days respectively. Results revealed a significant fall in the glycogen content, whereas, a marginal fall in the protein was also observed. The enzymatic activity of alkaline phosphatase and adenosine triphosphatase was depleted; on the contrary, there was a significant rise in the acid phosphatase and glucose-6-phosphatase pattern. A rise in the hepatic lipid peroxidation activity is a direct indication of oxidative damage resulting in free radical generation. The distribution of the metal by atomic absorption spectrophotometry revealed an increased concentration of beryllium in liver and kidney, followed by lung and uterus. The relative ability of three chelating agents to act as antagonists, for acute beryllium poisoning, have been examined in liver, kidney, lungs and uterus. The appreciable change in the beryllium concentration in various organs is duration dependent during the entire period being highly significant at 7 days regimen. Biochemical and distribution studies reveal that DPA + Se was the most effective therapeutic agent followed by DMPS + Se and GSH.