Optimization of amylase production using response surface methodology from newly isolated thermophilic bacteria.

Present study was aimed at screening and characterizing thermostable amylase-producing bacteria from water and sediment samples of unexplored hot spring of Tatta Pani Kotli Azad Kashmir. Four thermophilic isolates were characterized on morphological, biochemical, physiological basis and were authenticated by molecular analysis. By 16S rDNA sequencing, isolates were identified as Anoxybacillus mongoliensis (MBT001), Anoxybacillus flavithermus (MBT002), Bacillus (MBT004). Among all identified strains, MBT003 showed maximum homology with both Anoxybacillus mongoliensis and Anoxybacillus flavithermus. Amylase activity was analyzed qualitatively in starch agar and quantitatively by DNS method. The optimal enzyme production was observed and authenticated by Response Surface Methodology at 7 pH, 70 °C, 1.25% substrate concentration, 300 µL of inocula volume after 48 h of incubation. Optimum amylase activity (4.4 U/mL) and stability (3.3 U/mL) was observed with 1.5% soluble starch at 70 °C. Maximum activity (3.7 U/mL) and stability (1.5 U/mL) was found at pH 8. Enzyme activity was increased in the presence of MgSO4 and CaCl2. Amylase was stable with surfactants and commercial detergents for 30 min. Supplementation of the enzyme with commercial detergent improved the washing ability of the detergent. This investigation has revealed that these thermostable bacteria are excellent source of amylase which can be used commercially for generating economic activity on sustainable basis.

Diversity of culturable thermophilic bacteria from Tata Pani hotspring of Kotli Azad Jammu and Kashmir.

Hot water springs are unique areas populated by mesophiles, thermotolerant and hyperthermophiles. They are the source of diversity of thermophiles, mainly belonging to archaea and bacteria domains. The diversity of thermophiles gives an outline of the huge biological potential that can be exploited for industrial applications.To this end, this study was aimed to isolate and characterise the unexplored thermophilic microorganisms from hot water spring in Tatapani, Tehsil & District Kotli AJK, Pakistan. Around 10 bacterial isolates were identified using morphological, biochemical, physiological and molecular attributes. Sequencing of the 16S rDNA gene of the isolates followed by BLAST search revealed that the strain MBT008 has 100% similarity with Anoxybacilluskamchatkensis. MBT012 showed 99.57% similarity with A.mongoliensis, MBT014 was affiliated with A.tengchongensis with 99.43% similarity, MBT009 showed 99.83% homology with A.gonensis and MBT018, 98.70% similarity with A.karvacharensis. The presence of all this microbial diversity in one common source is of immense importance related to envioronmental and industrial aspects in general and extraction of thermostable enzymes from these thermophiles specifically opens new horizons in the field of industrial biotechnology. These thermophiles are revealing new capabilities and are being manipulated by biotechnologists in utilizing them in different unique ways.

Occurrence, influencing factors, toxicity, regulations, and abatement approaches for disinfection by-products in chlorinated drinking water: A comprehensive review.

Disinfection is considered as a vital step to ensure the supply of clean and safe drinking water. Various approaches are adopted for this purpose; however, chlorination is highly preferred all over the world. This method is opted owing to its several advantages. However, it leads to the formation of certain by-products. These chlorination disinfection by-products (DBPs) are genotoxic, carcinogenic and mutagenic. Still chlorination is being practiced worldwide. Present review gives insights into the occurrence, toxicity and factors affecting the formation of regulated (THMs, HAAs) and emerging DBPs (N-DBPs, HKs, HAs and aromatic DBPs) found in drinking water. Furthermore, remediation techniques used to control DBPs have also been summarized here. Key findings are: (i) concentration of regulated DBPs surpassed the permissible limit in most of the regions, (ii) high chlorine dose, high NOM, more reaction time (up to 3 h) and high temperature (up to 30 °C) enhance the formation of THMs and HAAs, (iii) high pH favors the formation of THMs while low pH is suitable of the formation of HAAs, (iv) high NOM, low temperature, low chlorine dose and moderate pH favors the formation of unstable DBPs (N-DBPs, HKs and HAs), (v) DBPs are toxic not only for humans but for aquatic fauna as well, (vi) membrane technologies, enhanced coagulation and AOPs remove NOM, (vii) adsorption, air stripping and other physical and chemical methods are post-formation approaches (viii) step-wise chlorination is assumed to be an efficient method to reduce DBPs formation without any treatment. Toxicity data revealed that N-DBPs are found to be more toxic than C-DBPs and aromatic DBPs than aliphatic DBPs. In majority of the studies, merely THMs and HAAs have been studied and USEPA has regulated just these two groups. Future studies should focus on emerging DBPs and provide information regarding their regulation.

Species-specific interaction of trihalomethane (THM) precursors in a scaled-up distribution network using response surface methodology (RSM).

Response surface methodology (RSM) with central composite design (CCD) was used to monitor and optimize species-specific interaction of trihalomethane (THM) precursors in a scaled-up distribution network (DN). Independent variables such as applied chlorine (Cl2), contact time (t), humic acid (HA) and bromide ions (Br-) were analyzed using full factorial CCD. Analysis of variance revealed a good agreement between experimental data and proposed a two-factor interaction model (p = .04, R2 = 0.7983). As a precursor, Cl- and Br- interaction with HA affected THMs' speciation. These precursor molecules were perceived least significant as discrete elements but HA: Br- and pH product significantly impacted total trihalomethane (TTHM) formation (r = 0.998, p = .007). This mutual interactive fraction was observed pH-dependent and influenced TTHM yield. Dibromochloromethane and bromoform formation was observed pH-dependent provided sufficient Br- in the system. Applied chlorine had significant (p = .01), while time had insignificant (p = .75) effect. Multiple response optimization suggested pH range between 6.0 and 7.6 and HA: Br- ratio between 1.3 and 5.9 were satisfactory for maintaining TTHM below ≤80 µg/L in DN with 0.88 desirability function (D). Their respective concentration may be minimized by changing precursor's individual concentration and possible combinations.

Monitoring of Coliforms and chlorine residual in water distribution network of Rawalpindi, Pakistan.

The present study was undertaken to examine the drinking water quality of Rawal Treatment Plant, Rawalpindi and its distribution network by collecting samples from eight different locations. The aim was to determine potential relationship between the presence of microorganisms and chlorine residual in the distribution network. Quantification of chlorine residual, turbidity, standard plate count (SPC), fecal and total coliforms by Most Probable Number (MPN) was performed. Three different forms of chlorine were measured at each sampling station such as free chlorine, residual chlorine, chloramines and total chlorine residual. A critical evaluation of data presented indicated that pH generally ranged from 7.02-7.30; turbidity varied from 0.34-2.79 NTU; conductivity fluctuated from 359-374 microS/cm; and TDS values were found to be ranging between 180-187 mg/l. Station # 7 was found to be most contaminated. The value of total chlorine was found to be 0.86 to1.7 mg/l at Station # 3 and 6, respectively. Highest standard plate count was 62 CFU/ml at Station # 7. Total coliforms were less than 1.1 MPN/100 ml at almost most of the stations except at Station # 3 where it was found to be greater than 23.0 MPN /100 ml. Overall aim of this study is to create awareness about contamination of drinking water in the water distribution networks and to make recommendations to provincial agencies such as EPA, CDA and WASA that regular monitoring should be carried out to ensure that the chlorine residual is available at consumer end.