Bioaugmentation: an approach to biological treatment of pollutants.

Industrial development and the associated generation of waste requires attention for their management, treatment, and reduction without further degrading the quality of life. Microbes and plant-based bioremediation approaches are some of the sustainable strategies for the biodegradation of harmful pollutants instead of chemical-based treatment. Bioaugmentation is one such approach where microbial strains with the ability to degrade the targeted pollutant are introduced in a polluted environment. Harnessing of microbes from various locations, especially from the site of contamination (indigenous microbes), followed by optimization of the strains, inoculum size, media, and genetic engineering of the microbes along with a combination of strategies such as bio stimulation, phytoremediation is being applied to increase the efficiency of bioaugmentation. Further, bioaugmentation is influenced by various factors such as temperature, the composition of the pollutant, and microbial inoculum which needs to be considered for maximum efficiency of the treatment process. It has numerous advantages such as low cost, sustainability, and easy handling of the contaminants however, the major limitation of bioaugmentation is to increase the survival rate of the microbes involved in remediation for a longer duration in such a highly toxic environment. The review discusses these various aspects of bioaugmentation in brief for its large-scale implementation to address the global issue of pollution and environment management.

Circulating serotypes and genotypes of dengue virus in North India: An observational study.

BACKGROUND OBJECTIVES: This study reports observation on circulating serotypes and genotypes of Dengue Virus in North India. METHODS: Serum samples were obtained from suspected cases of dengue referred to the virus diagnostic laboratory during 2014 to 2022. All samples were tested for anti-dengue virus IgM antibodies and NS1Ag by ELISA. NS1Ag positive samples were processed for serotyping and genotyping. RESULTS: Total 41,476 dengue suspected cases were referred to the laboratory of which 12,292 (29.6%) tested positive. Anti-Dengue Virus IgM antibodies, NS1Ag, both IgM and NS1Ag, were positive in 7007 (57.4%); 3200 (26.0%) and 2085 (16.0%) cases respectively. Total 762 strains were serotyped during 9-year period. DENV-1, DENV-2, DENV-3 and DENV-4 serotypes were found in 79 (10.37%), 506 (66.40%), 151 (19.82%) and 26 (3.41%) cases respectively. DENV-1, DENV-2 and DENV-3 were in circulation throughout. Total 105 strains were genotyped. Genotype IV of DENV-1 serotype was circulating till 2014 which was later replaced by genotype V. A distinct seasonality with increase in number of cases in post-monsoon period was seen. INTERPRETATION CONCLUSION: DENV-1, DENV-2 and DENV-3 were found to be in circulation in North India. Predominant serotype/genotype changed at times, but not at regular intervals.

Low prevalence of antimalarial-resistance mutations in India during 2014-15: Impact of combining first-line therapy with primaquine.

BACKGROUND: Antimalarial drug resistance surveillance and containment are crucial for countries aiming to eliminate malaria. Monitoring resistance evolution through studies before and after treatment policy changes is crucial. METHOD: A total of 939 P. falciparum-positive blood samples were collected between 2014 and 2015 across ten sites in India, categorized into four geographic clusters. PCR-amplified products were sequenced to identify point mutations at drug-resistance-conferring genes (Pfdhfr, Pfdhps, Pfmdr1, Pfk13). RESULT: Triple Pfdhfr mutants were found only in northeast India bordering Myanmar, while the wildtype was dominant in central India. Pfdhps wildtypes were prevalent in all areas, and no double mutants were found. Except in Northwest India, Pfmdr1 wildtype was dominant in all clusters. Nonsynonymous double mutations were only found in northwest India. Only synonymous mutations occurred in Pfk13. These were found in Central India at low frequency. The pattern of linkage disequilibrium and principal component analysis reflects low pressure for drug resistance and heterogeneity between the geographic clusters. CONCLUSION: Resistance levels were highest in Northeast India, close to the Myanmar border, where resistance is common. Primaquine has been widely used as a gametocidal and schizonticidal drug, has likely contributed to maintaining low drug resistance levels and preventing strong selection for resistance.

Effect of Entamoeba histolytica infection on gut microbial diversity and composition in diarrheal patients from New Delhi.

During amoebiasis, colonization of the gut by Entamoeba histolytica can lead to alterations of the host microbiota. In this study, we have compared the gut microbiota of patients of amoebiasis with healthy controls using 16S rRNA gene variable regions, (V1-V3) and (V3-V5), of the bacterial genome. From this 16S rRNA gene amplicon data, one paired-end and two single-end datasets were selected and compared by the number of OTUs obtained, sequence count, and diversity analysis. Our results showed that the V1-V3-paired-end dataset gave the maximum number of OTUs in comparison to the two single-end datasets studied. The amoebiasis samples showed a significant drop in richness in the alpha diversity measurements and lower intra group similarity compared to the healthy controls. Bacteria of genus Prevotella, Sutterella, and Collinsella were more abundant in healthy controls whereas Escherichia, Klebsiella, and Ruminococcus were more abundant in the E. histolytica-positive patients. All the healthy controls harbored bacteria belonging to Faecalibacterium, Prevotella, Ruminococcus, Subdoligranulum, and Escherichia genera while all the E. histolytica-positive patient samples contained genus Enterobacter. The compositional changes in the gut microbiome observed in our study indicated a higher prevalence of pathogenic bacteria along with a depletion of beneficial bacteria in E. histolytica-infected individuals when compared with healthy controls. These results underline the interplay between E. histolytica and the human gut microbiome, giving important inputs for future studies and treatments.

Microorganisms in biological control strategies to manage microbial plant pathogens: a review.

Chemical fertilizers and pesticides are an integral part of modern agriculture and are often associated with numerous environmental problems. Biological agents such as microorganisms can largely replace chemical fertilizers and pesticides. The proper use of selected microorganisms such as bacteria, fungi and viruses have several benefits for agriculture. These include a healthy soil microbiota, biological production of important compounds that promote plant health, and to be used as biocontrol agents (BCAs) that provide protection from plant pathogenic microorganisms. Scientists have found that several bacterial genera including Bacillus and Pseudomonas have antimicrobial activity against numerous pathogenic bacterial and fungal plant pathogens. Trichoderma, Aspergillus, and Penicillium are among the most common fungal genera used as BCAs against both bacterial and fungal plant pathogens. Several bacteriophages and mycoviruses are also found effective as BCAs against selective plant pathogens. Fusarium oxysporum is a commonly found microbial plant pathogen causing wilts and rots in plants. Overall, it can be concluded that the use of microbial BCAs is an effective practice against microbial plant pathogens.

Impact of Nutritional Profile on Gut Microbiota Diversity in Patients with Celiac Disease.

The pathogenesis of celiac disease (CD) is significantly influenced by gut microbiota. Daily nutritional profile influences the diversity of gut microbiota. This study was aimed to compare the abundance of gut microbiota in CD patients compared to normal control (NC), and to investigate the impact of nutritional factors on their fecal microbiota diversity. In this study, a selected panel of intestinal bacteria was assessed in 31 confirmed CD patients adhering to gluten-free diet (GFD) for more than 6 months and in 20 NC subjects. Stool samples were collected from each participant, DNA was extracted, and absolute quantitative real-time PCR (qPCR) was carried out. The gut microbiota including Bacteroidetes, Bifidobacterium, Clostridium, Staphylococcus, Enterobacteiaceae, Firmicutes, and Lactobacillus were assessed. The quantities of fruits, vegetables, meat, liquids, sugar and gluten-free candy/bread consumption were evaluated using a questionnaire. The proportion of Bifidobacterium, Firmicutes, and Lactobacillus in CD cases was significantly lower than NC (P < 0.005). Significant correlation coefficients between Bifidobacterium and Lactobacillus (P < 0.001), and also Firmicutes and Lactobacillus (P < 0.001) were recorded. Moreover, a significant association between medium amount of meat and bean consumptions and low abundance of Lactobacillus and Firmicutes (P = 0.024 and P = 0.027, respectively), and also high amount of bean consumptions and low abundance of Lactobacillus (P = 0.027) in CD were observed. The results showed that meat and bean consumptions could reduce the beneficial bacteria including Firmicutes and Lactobacillus in CD patients. Therefore, changes in the gut microbiota abundance may contribute to dietary changes and unimproved CD symptoms.

Monitoring the distribution pattern and invasion status of Ageratina adenophora across elevational gradients in Sikkim Himalaya, India.

Understanding the spread intensity and population dynamics of invasive plant species is a prerequisite for developing management strategies in the Himalayan Forest ecosystems that are experiencing an accelerated rate of climate change. Although there are studies on the occurrence of few invasive species in the Himalayan ecosystems, systematic information on their intensity of spread and species association is still missing. Considering existing data gaps, we aimed to assess the intensity of spread and distribution pattern of A. adenophora, one of the high-concern invasive species (HiCIS) of India that is causing havoc in the Himalayas, across an elevational gradient. Field data were collected in 2018 and 2021 in the Indian federal state of Sikkim, located in the Eastern Himalayas. We analyzed the population status and species association of A. adenophora along an elevational gradient ranging from > 600 m to 2700 m above sea level, which was divided into seven gradients of 300 m width, and each gradient was further randomly sampled. Overall, 81 species were present in association with A. adenophora, including 58 herbs, 19 shrubs, and 4 climbers, belonging to 30 families and 67 genera in the region. No other species continuously co-occurred along with A. adenophora throughout the elevation ranging from > 600 m to 2700 m. The species observed increased frequency (100%), density (40.51 ind./100 m2), and basal cover (11.25 cm2/m2) in the elevational gradient 1500-1800 m in 2018. In 2021, A. adenophora dominated the highest elevational gradient (< 2400-2700 m) with increased frequency (99.96%), density (58.41 ind./100m2), and basal cover (42.54 cm2/100m2), which demonstrated rapid invasion and improved plant health and reproductive vigor in comparison to the lower elevational gradient in Sikkim Himalaya. Despite being completely absent at the highest elevation (< 2400-2700 m), in 2018, it observed gregarious spread at the highest elevation in 2021, which is of serious concern to ecologists. The presence of the targeted species in all seven studied altitudinal gradients reflects stage III of the species invasion. An enormous shift in the distribution pattern along elevational gradients within a short time span is alarming for the Himalayan ecosystem since it is becoming a thriving habitat for invasive species owing to anthropogenic activity. We mapped the potential geographical extent using the species distribution model (SDM) and predicted the suitable habitat of distribution in Sikkim Himalaya. In order to curtail the spread and counteract the negative impact of this species on native vegetation in Sikkim Himalaya and ultimately reverse the process, local and regional initiatives for its biological control and management must be taken.

Role of extremophiles and their extremozymes in biorefinery process of lignocellulose degradation.

Technological advances in the field of life sciences have led to discovery of organisms that live in harsh environmental conditions referred to as extremophiles. These organisms have adapted themselves to thrive in extreme habitat giving these organisms an advantage over conventional mesophilic organisms in various industrial applications. Extremozymes produced by these extremophiles have high tolerance to inhospitable environmental conditions making them an ideal enzyme system for various industrial processes. A notable application of these extremophiles and extremozymes is their use in the degradation of recalcitrant lignocellulosic biomass and application in biorefineries. For maximum utilization of the trapped carbon source from this obstinate biomass, pretreatment is a necessary step that requires various physiochemical and enzymatic treatments. From search for novel extremophiles and extremozymes to development of various genetic and protein engineering techniques, investigation on extremozymes with enhanced stability and efficiency is been done. Since extremozymes are easily calibrated to work under such conditions, they have become an emerging topic in the research field of biofuel production. The review discusses the various extremozymes that play an important role in lignocellulose degradation along with recent studies on their molecular and genetic evolution for industrial application and production of biofuels and various value-added products.

Digitisation and realtime sharing of unified surveillance tool and clinicopathological data for efficient management of disease outbreaks.

Existing chronic co-morbidities in a population affect the clinical outcome of infectious diseases. In order to devise better management strategies at community level, patients related clinicopathological as well as local health care infrastructure data is required. The use of digitised, unified, multilingual surveillance tool may facilitate real-time sharing of clinicopathological data and better service delivery to the affected communities. In this article, we discuss that how digital collection, collation and sharing of health-related data may improve planning and management of disease outbreaks.

Severity of dengue illness and presence of anti DV IgG in serum of laboratory confirmed dengue cases.

BACKGROUND & OBJECTIVES: Presence of dengue is reported from India since 1960s. Secondary dengue infection may be more severe than primary, hence, distinction between primary and secondary dengue is essential. A way to detect secondary dengue is demonstration of anti DV IgG in patients' serum. In this study we explored the association of dengue severity with anti DV IgG positivity. METHODS: Laboratory confirmed cases of dengue (positive for anti DV IgM/ NS-1 Antigen/ DV -RNA), presenting to the hospital within 7 days of illness, were consecutively enrolled for a period of one month (September 1-30, 2018) and were tested for anti DV IgG in their serum. All PCR positive samples were serotyped. Cases positive for anti-dengue IgG were labeled as secondary cases. Clinical details were collected to assess the severity of illness. Association of dengue severity with anti DV IgG positivity was calculated. RESULTS: Of the 128 dengue positive cases, 89 (69.5%) were anti DV IgM positive, 72 (56.3%) were Dengue NS-1 positives and 37 (28.9%) were DV-RNA positive. Only 39 (30.5%) cases were having detectable anti-dengue IgG in their serum (secondary dengue). Anti-dengue IgM positivity was significantly higher in secondary dengue cases. No association of anti DV IgG positivity was seen with severity of dengue illness. INTERPRETATION & CONCLUSION: No association of IgG positivity with severity of illness was seen. D4 serotype is first time reported from Uttar Pradesh, India.

Human microbiome: an academic update on human body site specific surveillance and its possible role.

Human body is inhabited by vast number of microorganisms which form a complex ecological community and influence the human physiology, in the aspect of both health and diseases. These microbes show a relationship with the human immune system based on coevolution and, therefore, have a tremendous potential to contribute to the metabolic function, protection against the pathogen and in providing nutrients and energy. However, of these microbes, many carry out some functions that play a crucial role in the host physiology and may even cause diseases. The introduction of new molecular technologies such as transcriptomics, metagenomics and metabolomics has contributed to the upliftment on the findings of the microbiome linked to the humans in the recent past. These rapidly developing technologies are boosting our capacity to understand about the human body-associated microbiome and its association with the human health. The highlights of this review are inclusion of how to derive microbiome data and the interaction between human and associated microbiome to provide an insight on the role played by the microbiome in biological processes of the human body as well as the development of major human diseases.

Structure modeling and functional analysis of recombinant dextransucrase from Weissella confusa Cab3 expressed in Lactococcus lactis.

The dextransucrase gene from Weissella confusa Cab3, having an open reading frame of 4.2 kb coding for 1,402 amino acids, was amplified, cloned, and expressed in Lactococcus lactis. The recombinant dextransucrase, WcCab3-rDSR was expressed as extracellular enzyme in M17 medium with a specific activity of 1.5 U/mg which after purification by PEG-400 fractionation gave 6.1 U/mg resulting in 4-fold purification. WcCab3-rDSR was expressed as soluble and homogeneous protein of molecular mass, approximately, 180 kDa as analyzed by SDS-PAGE. It displayed maximum enzyme activity at 35°C at pH 5.0 in 50 mM sodium acetate buffer. WcCab3-rDSR gave Km of 6.2 mM and Vm of 6.3 µmol/min/mg. The characterization of dextran synthesized by WcCab3-rDSR by Fourier transform infrared and nuclear magnetic resonance spectroscopic analyses revealed the structural similarities with the dextran produced by the native dextransucrase. The modeled structure of WcCab3-rDSR using the crystal structures of dextransucrase from Lactobacillus reuteri (protein data bank, PDB id: 3HZ3) and Streptococcus mutans (PDB id: 3AIB) as templates depicted the presence of different domains such as A, B, C, IV, and V. The domains A and B are circularly permuted in nature having (ß/α)8 triose phosphate isomerase-barrel fold making the catalytic core of WcCab3-rDSR. The structure superposition and multiple sequence alignment analyses of WcCab3-rDSR with available structures of enzymes from family 70 GH suggested that the amino acid residue Asp510 acts as a nucleophile, Glu548 acts as a catalytic acid/base, whereas Asp621 acts as a transition-state stabilizer and these residues are found to be conserved within the family.

The family 6 Carbohydrate Binding Module (CtCBM6) of glucuronoxylanase (CtXynGH30) of Clostridium thermocellum binds decorated and undecorated xylans through cleft A.

CtCBM6 of glucuronoxylan-xylanohydrolase (CtXynGH30) from Clostridium thermocellum was cloned, expressed and purified as a soluble ~14 kDa protein. Quantitative binding analysis with soluble polysaccharides by affinity electrophoresis and ITC revealed that CtCBM6 displays similar affinity towards decorated and undecorated xylans by binding wheat- and rye-arabinoxylans, beechwood-, birchwood- and oatspelt-xylan. Protein melting studies confirmed thermostable nature of CtCBM6 and that Ca(2+) ions did not affect its structure stability and binding affinity significantly. The CtCBM6 structure was modeled and refined and CD spectrum displayed 44% ß-strands supporting the predicted structure. CtCBM6 displays a jelly roll ß-sandwich fold presenting two potential carbohydrate binding clefts, A and B. The cleft A, is located between two loops connecting ß4-ß5 and ß8-ß9 strands. Tyr28 and Phe84 present on these loops make a planar hydrophobic binding surface to accommodate sugar ring of ligand. The cleft B, is located on concave surface of ß-sandwich fold. Tyr34 and Tyr104 make a planar hydrophobic platform, which may be inaccessible to ligand due to hindrance by Pro68. Site-directed mutagenesis revealed Tyr28 and Phe84 in cleft A, playing a major role in ligand binding. The results suggest that CtCBM6 interacts with carbohydrates through cleft A, which recognizes equally well both decorated and un-decorated xylans.

Titers of anti-tissue transglutaminase antibody correlate well with severity of villous abnormalities in celiac disease.

GOALS: We reviewed our celiac disease (CeD) database to study if anti-tissue transglutaminase (tTG) antibody (ab) titers correlate with severity of villous abnormalities in Indian patients and to find out a cutoff value of anti-tTG ab fold-rise, which could best predict CeD. BACKGROUND: Guidelines for diagnosing CeD suggest that biopsy could be avoided in some patients with high anti-tTG ab titer. STUDY: We reviewed a cohort of 366 anti-tTG ab-positive individuals in whom duodenal biopsies were performed. Anti-tTG ab was obtained before initiation of gluten-free diet. Anti-tTG ab results were expressed in terms of fold-rise by calculating ratio of observed values with cutoff value. CeD was diagnosed if in addition to positive serology, patients had villous atrophy (>Marsh grade 2) and unequivocal response to gluten-free diet. RESULTS: The mean anti-tTG fold-rise in groups with Marsh grade ≤2 was 2.6 (±2.5), grade 3a was 4.0 (±3.9), 3b was 5.7 (±5.1), and 3c was 11.8 (±8.0). The positive likelihood ratio for diagnosing CeD was 15.4 and 27.4 at 12- and 14-fold-rise of anti-tTG ab titer, respectively. The positive predictive value of diagnosis of CeD was 100% when anti-tTG ab titer was 14-fold higher over the cutoff value. Fifty-seven (43.9%) individuals with anti-tTG titer rise <2-fold high also had CeD. CONCLUSIONS: As severity of villous abnormality increases, titer of anti-tTG also rises. Presence of villous atrophy can be predicted at very high anti-tTG ab titer. In contrast to emerging belief, mucosal biopsies should be performed even if anti-tTG ab titer is <2 times, because many patients with CeD have low titers.

Antipsychotic activity of standardized Bacopa extract against ketamine-induced experimental psychosis in mice: Evidence for the involvement of dopaminergic, serotonergic, and cholinergic systems.

CONTEXT: Schizophrenia is a chronic disabling psychiatric disorder affecting 1% of the population worldwide. Due to the adverse effects of available antipsychotic medications, recent investigations have focused on the search for well-tolerated, safe molecules from natural resources to control the severity and progression of schizophrenia. OBJECTIVE: To screen the standardized extract of Bacopa monniera Linn. (Scrophulariaceae) (BM) for its antipsychotic potential in the ketamine-induced psychosis model with mice. MATERIALS AND METHODS: Graded dose of BM (40, 80, and 120 mg/kg, p.o.) were given to the mice 1 h prior to ketamine administration and tested for positive symptoms and cognitive deficits. A chronic ketamine treatment regimen was used to study the effect of BM on negative symptoms such as immobility enhancement. Each mouse was used once for the behavioral studies. RESULTS: BM reduced ketamine-induced hyperactivity with an EC50 value of 76.60 mg/kg. The 80 mg/kg dose was used for all other behavior analysis. Pretreatment with BM at 80 mg/kg showed two-fold increases in transfer latency time (TLT) in passive avoidance task. Chronic BM pretreatment (80 mg/kg p.o. daily × 10 d) ameliorated the ketamine-induced enhanced immobility effect by 21% in the forced swim test. BM treatment reversed ketamine-induced increase in monoamine oxidase activity in both cortex and striatum and normalized the acetylcholinesterase activity and the glutamate levels in the hippocampus. DISCUSSION AND CONCLUSION: Overall our findings suggest that BM possesses antipsychotic properties which might be due to its modulatory action on dopamine, serotonin, and glutamate neurotransmission.

Molecular characterization of circulating pandemic strains of influenza A virus during 2012 to 2013 in Lucknow (India).

The pandemic H1N1 strain of Influenza A virus [A(H1N1)pdm09] is now well adapted in human populations. However, it is still causing sporadic outbreaks worldwide with different severity. The present study was planned to understand the genetic diversity (based on the HA1 gene) of influenza A(H1N1)pdm09 strains circulating during the post pandemic period. The HA1 gene was selected because the HA1 protein is immunogenic, functions as a receptor binding site and indirectly affects transmission and pathogenicity of virus. A total of 2,818 cases were enrolled. Nasal/throat swabs from all cases were tested by one-step real time PCR for detection of influenza virus types and subtypes according to the CDC protocol. Of these, 134 cases were A(H1N1)pdm09 positive, 34 of which were screened for HA1 gene (position 434-905) sequencing (Big-Dye terminator using 3130 ABI, Genetic analyzer). Molecular and phylogenetic analysis was performed using PhyML approach (v. 3.0). All A(H1N1)pdm09 positive and negative cases were clinically characterized. Phylogentically, all Lucknow strains (n = 33) except one fall with the clade seven reference strain. One strain showed 99.9% similarities with clade one reference strain A/California/07/2009. In mutational analysis, 33 strains had the S220T mutation, which is at an antigenic site and characteristic of clade seven along with few minor mutations; K180I/T/Q, V190I, S200P, S202T, A203T, A214T, S220T, V251I, and A273T. These results suggest that clade seven was the most widely circulating clade in Lucknow and A(H1N1)pdm09 cases showed mild clinical symptoms as compared to A(H3N2) or influenza B cases.

[In silico structural characterization and molecular docking studies of first glucuronoxylan-xylanohydrolase (Xyn30a) from family-30 glycosyl hydrolase (GH30) from Clostridium thermocellum].

CtXynGH30 is a carbohydrate active modular enzyme and component of cellulosome of Clostridium thermocellum. The full length CtXynGH30 contains an N-terminal catalytic module named as Xyn30A and a family 6 carbohydrate binding module (CBM6) at C-terminuis. Xyn30A was modeled by computer program Modeller9v8 taking crystal structure of XynC from B. subtilis as a template to generate the molecular model. Model refinement was done using energy minimization by implementing steepest descent algorithm with GROMOS96 43al force field. Quality assessment by Ramachandran plot showed that 91% amino acids lie in most favourable region and 9% in additional allowed region. Structural analysis depicted that Xyn30A has a (beta/alpha)8 barrel fold. Ad- ditionally, it had a beta-strand rich structure called 'side beta-structure' attached with main catalytic core. Structural superimposition reflected that Glu136 act as a catalytic acid/base while Glu225 act as a catalytic nucleophile. Multiple sequence alignment showed that these catalytic residues are conserved within the family. The docking results showed that these residues display polar interaction with linear and substituted xylo-oligosaccharides. The binding interaction of ligands depicted that aromatic amino acids Trp81, Tyr139, Trp143, Phe172, His198, Tyr200, Tyr227, Trp264 and Tyr265 create binding site pocket around the active site. We report overall structural feature, conserved active site residues and enzymeligand docking of first glucuronoxylan-xylanohydrolase (Xyn30A) of family 30 glycosyl hydrolase (GH30) from Clostridium thermocellum.

A comparative study of magnesium sulfate vs dexmedetomidine as an adjunct to epidural bupivacaine.

BACKGROUND AND AIMS: This prospective, randomized, double-blind study was undertaken to establish the effect of addition of magnesium or dexmedetomidine, as an adjuvant, to epidural bupivacaine in lower limb surgeries. MATERIALS AND METHODS: One hundred and twenty ASA (American Society of Anesthesiologists) class I and II patients undergoing lower limb surgeries were enrolled to receive either magnesium sulfate (Group M) or dexmedetomidine (Group D) along with epidural bupivacaine for surgical anesthesia. All the study subjects received an epidural anesthesia with 14 ml of 0.5% bupivacaine along with either MgSO4 50 mg (Group M) or dexmedetomidine 0.5 µg/kg (Group D) or saline (Group C). The onset of motor and sensory block, duration of block, hemodynamic parameters, and any adverse events were monitored. RESULTS: Analgesia in the postoperative period was better in Group D, duration of sensory and motor blockade was significantly prolonged in Group D and incidence of sedation was more in Group D. CONCLUSION: Hence, addition of dexemedetomidine to epidural bupivacaine can be advantageous with respect to increased duration of motor and sensory blockade and arousable sedation.

Statistical optimization of cellulase production from Bacillus sp. YE16 isolated from yak dung of the Sikkim Himalayas for its application in bioethanol production using pretreated sugarcane bagasse.

Cellulolytic bacteria were isolated from yak dung samples collected from different habitats of Sikkim, India. Isolate YE16 from the Yumthang Valley sample showed highest enzyme activity of 7.68 U/mL and was identified as Bacillus sp., which has a sequence similarity of 96.15% with B. velezensis. One factor at a time (OFAT) analysis revealed that an acidic pH of 5 with 37 °C temperature was optimum for maximum enzyme production after 36 hrs of incubation (13.88 U/mL), which was further increased after statistical optimization (34.70 U/mL). Media optimization based on response surface methodology predicted that Carboxymethyl cellulose (CMC) and MgSO4 at concentrations of 30 g/L and 0.525 g/L, respectively, at pH 5.5 to show CMCase activity of 30.612 U/mL, which was consistent with the observed value of 30.25 U/mL and confirmed the model. The crude enzyme also efficiently hydrolyzed alkaline pretreated sugarcane bagasse, releasing 7.09 g/L of glucose equivalent with an ethanol production of 3.05 g.