Search results for: anaerobic digestion

Authors: Kumaran Narayanan, Pei-Sheng Liew

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This paper adapts the bacteriophage N15 protelomerase enzyme to assemble linear chromosomes as vectors for gene expression in human cells. Phage N15 has the unique ability to replicate as a linear plasmid with telomeres in E. coli during its prophage stage of life-cycle. The virus-encoded protelomerase enzyme cuts its circular genome and caps its ends to form hairpin telomeres, resulting in a linear human-chromosome-like structure in E. coli. In mammalian cells, however, no enzyme with TelN-like activities has been found. In this work, we show for the first-time transfer of the protelomerase from phage into human and mouse cells and demonstrate recapitulation of its activity in these hosts. The function of this enzyme is assayed by demonstrating cleavage of its target DNA, followed by detecting telomere formation based on its resistance to recBCD enzyme digestion. We show protelomerase expression persists for at least 60 days, which indicates limited silencing of its expression. Next, we show that an intact human β-globin gene delivered on this linear chromosome accurately retains its expression in the human cellular environment for at least 60 hours, demonstrating its stability and potential as a vector. These results demonstrate that the N15 protelomerse is able to function in mammalian cells to cut and heal DNA to create telomeres, which provides a new tool for creating novel structures by DNA resolution in these hosts.

Keywords: chromosome, beta-globin, DNA, gene expression, linear vector

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Authors: Mallika Supa-Aksorn, Buaream Maneewan, Jiraporn Rojtinnakorn

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For animals, trypsin and chymotrypsin are the 2 proteases that play the important role in protein digestion and involving in growth rate. In many animals, these two enzymes are indicated as growth parameter by feed. Although enzyme assay at optimal condition is significant for its accuracy activity determination. There is less report of trypsin and chymotrypsin. Therefore, in this study, optimization of pH and temperature for trypsin (T) and chymotrypsin (C) in economic species; i.e. Nile tilapia (Oreochromis niloticus), sand goby (Oxyeleotoris marmoratus), giant freshwater prawn (Macrobachium rosenberchii) and native chicken (Gallus gallus) were investigated. Each enzyme of each species was assaying for its specific activity with variation of pH in range of 2-12 and temperature in range of 30-80 °C. It revealed that, for Nile tilapia, T had optimal condition at pH 9 and temperature 50-80 °C, whereas C had optimal condition at pH 8 and temperature 60 °C. For sand goby, T had optimal condition at pH 7 and temperature of 50 °C, while C had optimal condition at pH 11 and temperature of 70-75 °C. For juvenile freshwater prawn, T had optimal condition at pH 10-11 and temperature of 60-65 °C, C had optimal condition at pH 8 and temperature of 70°C. For starter native chicken, T has optimal condition at pH 7 and temperature of 70 °C, whereas C had o optimal condition at pH 8 and temperature of 60°C. This information of optimal conditions will be high valuable in further for, actual enzyme measurement of T and C activities that benefit for growth and feed analysis.

Keywords: trypsin, chymotrypsin, Oreochromis niloticus, Oxyeleotoris marmoratus, Macrobachium rosenberchii, Gallus gallus

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Authors: Li Bixia, Ren Shouwen, Fu Yanfeng, Tu Feng, Xiaoming Fang, Xueming Wang

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The utilization of dietary crude fiber in different breed pigs is not the same. Su-shan pigs are a new breed formed by crossing Taihu pigs and Yorkshire pigs. In order to understand the resistance of Su-shan pigs to dietary crude fiber, 150 Su-shan pigs with 60 kg of average body weight and similar body conditions were allocated to three groups randomly, and there are 50 pigs in each group. The percentages of dietary crude fiber were 8.35%, 9.10%, and 11.39%, respectively. At the end of the experiment, 15 pigs randomly selected from each group were slaughtered. The results showed as follows: average daily gain of the 9.10% group was higher than that of the 8.35% group and the 11.39% group; there was a significant difference between the 9.10% group and the 8.35% group (p < 0.05. Levels of urea nitrogen, total cholesterol and high density lipoprotein in the 9.10% group were significantly higher than those in the 8.35% group and the 11.39% group (p < 0.05). Ratios of meat to fat in the 9.10% group and the 11.39% group were significantly higher than that in the 8.35% group (p < 0.05). Lean percentage of 9.10% group was higher than that of 8.35% group and 11.39% group, but there was no significant difference in three groups (p > 0.05). The weight of small intestine and large intestine in the 11.39% group was higher than that in the 8.35% group, and the 9.10% group and the difference reached a significant level (p < 0.05). In conclusion, increasing dietary crude fiber properly could reduce fat percentage, and improve the ratio of meat to fat of finishing Su-shan pigs. The digestion and metabolism of dietary crude fiber promoted the development of stomach and intestine of finishing Su-shan pig.

Keywords: Su-shan pigs, dietary crude fiber, growth performance, serum biochemical indexes

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Authors: Zasha Romero, Roberto Trevino, Lin Wang, Elizabeth Alanis, Jesus Cuellar

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Background: There is a strong relationship between low cardiorespiratory fitness (CRF) and high adiposity levels. The purpose of this study was to assess the effects of the Bienestar/Neema Coordinated School Health Program (BN CSHP) on the CRF of preschool children. Methods: This is a randomized cluster trial conducted in preschools of two school districts located along the Texas-Mexico border. Of 48 eligible schools, 28 were randomly selected (intervention, n=14; control, n=14). Family demographics and household health characteristics were collected from parents. CRF, as measured by the Progressive Anaerobic Capacity Endurance Run (PACER) fitness test, was collected from the children. A generalized linear mixed model (GLMM) was used to analyze the data. Results: Family demographics, household health characteristics, and children’s weight, obesity prevalence, and sedentary activity were similar among both treatment groups. After adjusting for covariates, the number of laps run by children in the control group increased by 23% (CI: -5% to 60%) per each data collection period compared with 53% (CI: 7% to 119%) in the intervention group. Conclusions: Children in the BN CSHP, compared to those in the control group, had a significantly higher increase in their CRF. This finding is important because of the health benefits of CRF in children.

Keywords: coordinated school health program, cardiorespiratory fitness, obesity, border health, preschool, physical education, movement

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Authors: S. Hallal, Z. Chami, A. Hadji-Lehtihet, S. Sokhal-Boudella, A. Berhoune, L. Yargui

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Gaucher disease is the most common lysosomal storage in our population, it is due to a deficiency of β –glucosidase acid. The enzyme deficiency causes a pathological accumulation of undegraded substrate in lysosomes. This metabolic overload is responsible for a multisystemic disease with hepatosplenomegaly, anemia, thrombocytopenia, and bone involvement. Neurological involvement is rare. The laboratory diagnosis of Gaucher disease consists of phenotypic diagnosis by determining the enzymatic activity of β - glucosidase by fluorimetric method, a study by genotypic diagnosis in the GBA gene, limiting the search recurrent mutations (N370S, L444P, 84 GG); PCR followed by an enzymatic digestion. Abnormal profiles were verified by sequencing. Monitoring of treated patients is provided by the determination of chitotriosidase. Our experience spaning a period of 6 years (2007-2014) has enabled us to diagnose 78 patients out of a total of 328 requests from the various departments of pediatrics, internal medicine, neurology. Genotypic diagnosis focused on the entire family of 9 children treated at pediatric CHU Mustapha, which help define the clinical form; or 5 of them had type III disease, carrying the L444P mutation in the homozygous state. Three others were composite (N370/L444P) (N370S/other unintended mutation in our study), and only in one family no recurrent mutation has been found. This molecular study permits screening of heterozygous essential for genetic counseling.

Keywords: Gaucher disease, mutations, N370S, L444P

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Authors: Ehiedu Philomina Kika, Jessica Chinonso Ehilegbu

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The exploration and production of fossil fuel particularly crude oil has led to some serious environmental damage in some oil producing communities like the Bodo Community who rely heavily on their aquatic environment for food and water. This study was therefore carried out to investigate the level of some heavy metals in water and Callinectes amnicola (Swimming Crab) in the month of August, September and October from Bodo creek, Rivers State, Nigeria. The physico-chemical parameters of the water were also analyzed in-situ. The levels of heavy metals, Lead (Pb), Cadmium (Cd), Chromium (Cr), Zinc (Zn), Copper (Cu) were analyzed in water and in Callinectes amnicola (Swimming Crab), using Atomic Absorption Spectrophotometer (AAS) after acid digestion. For the concentration of heavy metals in water, Pb ranged from 0.103 - 0.791 mg/l, Zn 0.0025 - 0.342 mg/l, Cr < 0.001 - 0.304 mg/l, Cd 0.011 - 0.116 mg/l and Cu <0.001 - 0.079 mg/l. For the concentration of heavy metals in Callinectes amnicola (Swimming Crab), the level of Pb ranged from 0.359 - 0.849 mg/l, Zn 0.134 - 0.342 mg/l, Cd 0.053 - 0.103 mg/l, Cr < 0.001 - <0.001 mg/l, Cu < 0.001 - 0.131 mg/l. The concentrations of Pb, Cd and Cr for all water and crab samples collected from the various stations were higher than permissible level suggesting serious anthropogenic influence. Thus, precaution needs to be taken to prevent further contamination and adequate purification measures need to be put in place. Therefore, there should be periodic environmental pollution monitoring, for assessment and awareness especially with regards heavy metal.

Keywords: Bodo creek, crude oil, heavy metal, swimming crab

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Authors: Michael Graz, Andrew Shearer, Gareth Evans

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Manipulation of rumen bacteria is receiving increasing attention as a way of controlling greenhouse gas (GHG) emissions that are generated by the agricultural sector. Feed supplementation in particular is one of the ways in which this drive is being addressed, in particular with reference to livestock-generated GHG emissions. A blend of naturally occurring chemical extracts obtained from garlic and bitter orange extracts has been identified as a natural, sustainable and non-antibiotic based way of reducing methane production by ruminant livestock. In the current study, the acceptability and impact of this blend of natural extracts on feed rations of beef cattle was trialed in vivo on a commercial farm in Europe. Initial findings have demonstrated acceptable palatability, with all animals accepting the feed supplement into their ration both when it was mixed into the total daily ration and when used as a part of their high energy rations. Measurement of the impact of this feed supplement on productivity weight gain and milk quality is ongoing. In conclusion, this field study confirmed the palatability of the combination of garlic and citrus extracts and hence pointed to possibility of the extract blend to improve digestion, enhance body energy retention and limit CH4 formation in relation to feed intake.

Keywords: citrus, garlic, methane reduction, palatability, ruminants

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Authors: Mahesh Kumar Farejiya, Anil Kumar Dikshit

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The toxic metal contamination and their biomagnification in marine fishes is a serious public health concern specially, in the coastal areas and the small islands. In the present study, concentration of toxic heavy metals like zinc (Zn), cadmium (Cd), lead (Pb), nickel (Ni), cobalt (Co), chromium (Cr) and mercury (Hg) were determined in the tissues of tunas (T. albacores) caught from the area near to Lakshdweep Islands. The heavy metals are one of the indicators for the marine water pollution. Geochemical weathering, industrialization, agriculture run off, fishing, shipping and oil spills are the major pollutants. The presence of heavy toxic metals in the near coastal water fishes at both western coast and eastern coast of India has been well established. The present study was conducted assuming that the distant island will not have the metals presence in a way it is at the near main land coast. However, our study shows that there is a significant amount of the toxic metals present in the tissues of tuna samples. The gill, lever and flash samples were collected in waters around Lakshdweep Islands. They were analyzed using ICP–AES for the toxic metals after microwave digestion. The concentrations of the toxic metals were found in all fish samples and the general trend of presence was in decreasing order as Zn > Al > Cd > Pb > Cr > Ni > Hg. The amount of metals was found to higher in fish having more weight.

Keywords: toxic metals, marine tuna fish, bioaccumulation, biomagnifications

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Authors: Debasmita Dubey, Rajesh Kumar Meher, Smruti Pragya Samal, Pradeep Kumar Naik

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Background: To determine antioxidant properties and anticancer activity by ROS and mitochondrial transmembrane potential mediated apoptosis against MCF7, MDA-MB-231, cell line. Methods: Leaf sample was extracted using methanol by microwave digestion technique. The antioxidant properties of the methanolic extract were determined by a DPPH scavenging assay. In vitro anticancer activity, mitochondrial transmembrane potential, apoptosis activity and DNA fragmentation study, as well as intracellular ROS activity of most potential leaf extract, were also determined by using the MDA-MB-231cell line. In vivo animal toxicity study was carried out using mice model. Results: Methanolic leaf extract has shown the highest antioxidant, as well as anticancer activity, is based on the assay conducted. For the identification of active phytochemicals from methanolic extract, High-resolution mass spectroscopy-LCMS was used. In vitro cytotoxicity study against MCF-7 and MDA-MB-231 cell line and IC 50 value was found to be 37.5µg/ml. From histopathological studies, no toxicity in liver and kidney tissue was identified. Conclusion: This plant tuber can be used as a regular diet to reduce the chance of breast cancer. Further, more studies should be conducted to isolate and identify the responsible compound.

Keywords: human breast adenocarcinoma, ROS, mitochondrial transmembrane, apoptosis

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Authors: Sarisha Devnath, Oluwatosin A. Ijabadeniyi

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In recent years milk contamination has become a major problem in households; due to the likely occurrence of bacteria, even after the milk has been processed. One such genus of bacteria causing unwanted growth is Bacillus. This research project looks at the presence of spore formers in processed milk from household refrigerators and the effect of pasteurization and high temperature on Bacillus spores activation. 24 samples each of UHT milk and pasteurised milk from 24 households were sampled for the presence of spore formers. While anaerobic spore formers were not found in any of the samples, the average aerobic spore formers in UHT milk and pasteurized milk however were 5.77 cfu/ml and 5.88 cfu/ml respectively. After sequencing, it was detected that the mixed culture contained Bacillus cereus, for both pasteurised and UHT milk samples. For the activation study, raw milk samples were collected and subjected to four different temperatures; 65˚C, 72˚C, 80˚C, 100˚C respectively. Samples were stored for 7 days at 5˚C and 10˚C and analysed daily. The average aerobic spore formers in raw milk for samples stored at 5˚C range between 4.67-6.00 cfu/ml while it ranges between 4.84-6.00 cfu/ml at 10˚C, signifying that the high temperatures could have resulted in germination of dominant spores. Statistical analysis conducted on these results indicated a significant difference between the numbers of colonies present at the different treatment temperatures the bacterium was exposed to. This work showed that household milk may constitute public health risk furthermore; pasteurization and higher temperatures may not be effective to remove aerobic spore formers because of Bacillus spores activation.

Keywords: sporeformers, bacillus, spores, activation, milk

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Authors: Jaruwan Chutrtong

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Yoghurt is a fermented milk product. The process of making yogurt involves fermenting milk with live and active bacterial cultures by adding bacteria directly to the dairy product. It is usually made with a culture of Lactobacillus sp. (L. acidophilus or L. bulgaricus) and Streptococcus thermophilus. Many people like to eat it plain or flavored and it's also use as ingredient in many dishes. Yogurt is rich in nutrients including the microorganism which have important role in balancing the digestion and absorption of the boy.Consumers will benefit from lactic acid bacteria more or less depending on the amount of bacteria that lives in yogurt while eating. When purchasing yogurt, consumers should always check the label for live cultures. Yoghurt must keep in refrigerator at 4°C for up to ten days. After this amount of time, the cultures often become weak. This research studied freezing dry yogurt storage by monitoring on the survival of microorganisms when stored at different temperatures. At 300°C, representative room temperature of country in equator zone, number of lactic acid bacteria reduced 4 log cycles in 10 week. At 400°C, representative temperature in summer of country in equator zone, number of lactic acid bacteria also dropped 4 log cycle in 10 week, similar as storage at 300°C. But drying yogurt storage at 400°C couldn’t reformed to be good character yogurt as good as storage at 400°C only 4 week storage too. After 1 month, it couldn’t bring back the yogurt form. So if it is inevitable to keep yogurt powder at a temperature of 40°C, yoghurt is maintained only up to 4 weeks.

Keywords: dynamic, dry yoghurt, storage, temperature

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Authors: Pinar Ercan, Sedef N. El

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The goals of this study were to determine the total anthocyanin and procyanidin contents and their in vitro bioaccessibilities of apple, red grape and cinnamon by a static in vitro digestion method reported by the COST FA1005 Action INFOGEST, as well as in vitro inhibitory effects of these food samples on starch and lipid digestive enzymes. While the highest total anthocyanin content was found in red grape (164.76 ± 2.51 mg/100 g), the highest procyanidin content was found in cinnamon (6432.54±177.31 mg/100 g) among the selected food samples (p<0.05). The anthocyanin bioaccessibilities were found as 10.23±1 %, 8.23±0.64 %, and 8.73±0.70 % in apple, red grape, and cinnamon, respectively. The procyanidin bioaccessibilities of apple, red grape, and cinnamon were found as 17.57±0.71 %, 14.08±0.74 % and 18.75±1.49 %, respectively. The analyzed apple, red grape and cinnamon showed the inhibitory activity against α-glucosidase (IC50 544.27±21.94, 445.63±15.67, 1592±17.58 μg/mL, respectively), α-amylase (IC50 38.41±7.26, 56.12±3.60, 3.54±0.86 μg/mL, respectively), and lipase (IC50 52.65±2.05, 581.70±54.14, 49.63±2.72 μg/mL, respectively). Red grape sample showed the highest inhibitory activity against α-glucosidase, cinnamon showed the highest inhibitory activity against α-amylase and lipase according to IC50 (concentration of inhibitor required to produce a 50% inhibition of the initial rate of reaction) and Catechin equivalent inhibition capacity (CEIC50) values. This study reported that apple, grape and cinnamon samples can inhibit the activity of digestive enzymes in vitro. The consumption of these samples would be used in conjunction with a low-calorie diet for body weight management.

Keywords: anthocyanin, α-amylase, α-glucosidase, lipase, procyanidin

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Authors: Hassan Nabipour Afrouzi, Naser Mahmoudnia

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Broiler production has increased rapidly in tropical and subtropical regions in the past and sustained growth is forecast for the future. One of the greatest challenges to efficient production in these regions is reduced performance from warm and hot weather conditions. There are many ways to decrease these detrimental effects of heat on broiler chickens. One way is to supplement broiler diet with betaine added to feed or drinking water. A review of the results of this study suggest that betaine supplement was effective to significantly improve body weight and feed conversion ratio at the initial stages of growth but not in the finisher stages (P<0/05). It was also demonstrated that the use of betaine significantly reduced the percentage of abdominal meat and the percentage of breast meat (P<0/05), but had no effect on other carcass compositions. Betaine may improve the digestibility of specific nutrients. Betaine, as a methyl donor provides labile methyl groups for the synthesis of several metabolically active substances such as creatine and carnitine. Oil in a broiler diet is known to promote a response to dietary betaine supplements, that is, chicks have a higher demand for betaine with a high fat diet. This study implies that betaine supplement may stimulate protection of intestinal epithelium against osmotic disturbance, improve digestion and absorption conditions of the gastrointestinal tract and promote amended use of nutrients.

Keywords: heat stress, betaine, performance, broiler‚ growth

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Authors: Mian Adnan Kakakhel, NIshita Narwal, Majid Rasta, Shi Xiaotao

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The widespread use of nanoparticles means that they are significantly increasing in the aquatic ecosystem, where they are likely to pose threat to aquatic organism. In particular, the influence of nanoparticles exposure combined with varying water velocities on fish behavior remain poorly understood. Emerging evidences suggested a probable correlation between fish swimming behavior and gut bacterial dysbiosis. Therefore, the current study aimed to investigate the effects of nanomaterials in different water velocities on fish gut bacterial composition, which in results change in fish swimming behavior. The obtained findings showed that the contamination of nanoparticles was reduced as the velocity increased. However, the synergetic effects of nanoparticles and water velocity significantly (p < 0.05) decreased the bacterial composition, which plays a critical role in fish development, metabolism, digestion, enzymes production, and energy production such as Bacteroidetes and Firmicutes. This group of bacterial also support fish in swimming behavior by providing them a significant energy during movement. The obtained findings of this study suggested that the presence of nanoparticles in different water velocities have had a significant correlation with fish gut bacterial dysbiosis, as results the gut dysbiosis had been linked to the change in fish behavior. The study provides an important insight into the mechanisms by which the nanoparticles possibly affect the fish behavior.

Keywords: water velocities, fish behavior, gut bacteria, secondary metabolites, regulation

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Authors: Roni Rachel Mendelson, Caileigh Pudela

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Bacillus cereus is a toxin-producing, facultatively anaerobic gram-positive bacterium that is widely distributed environmentally. It can quickly multiply at room temperature with an abundantly present preformed toxin. When ingested, this toxin can cause gastrointestinal illness, which is the commonly known manifestation of the disease. Bacillus cereus sepsis is a disease that is mostly concerning in the population of the immunocompromised patients. One of them is acute lymphoblastic leukemia’s patients during induction. Pediatric acute lymphoblastic leukemia is a common pediatric hematologic malignancy. It is characterized by the rapid proliferation of poorly differentiated lymphoid progenitor cells inside the bone marrow. We present here a 21-month-old boy undergoing induction chemotherapy for acute lymphoblastic leukemia who developed bacillus sepsis bacteremia and, as a result, multi organ failure leading to seizures and multiple strokes. Our case report highlights the extensive overall and neurological damage that can be caused because of bacillus cereus bacteremia, which can lead to higher mortality rate and decreased in survivorship in a highly curable disease. It is very subtle and difficult to recognize and appears to be deteriorating extremely fast. There should be a low threshold for work up and empiric coverage for neutropenic patients during acute lymphoblastic leukemia induction therapy.

Keywords: acute lymphoblastic leukemia, bacillus cereus, immunocompromised, sepsis

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Authors: Jeonghyun Chang, Taegeun Lee, Heungsup Sung, Yoon-Seon Lee, Youn-Jung Kim, Mi-Na Kim

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Background: Safety device has been applied to improve safety and performance of blood culture practice. BD vacutainer® Safety-Lokᵀᴹ blood collection sets with pre-attached holder (Safety-Lok) (BD, USA) was evaluated in the emergency room (ER) of a tertiary care hospital. Methods: From April to June 2017, interns and nurses in ER were surveyed for blood culture practices with a questionnaire before and after 2 or 3 weeks of experience of Safety-Lok. All of them participated in exercise workshop for 1 hour combined with video education prior to the initial survey. The blood volume, positive and contamination rates of Safety-Lok-drawn (SD) blood cultures were compared to those of overall blood cultures. Results: Eighteen interns and 30 nurses were enrolled. As a result of the initial survey, interns had higher rates of needlestick incidence (27.8%), carriage of the blood-filled syringe with needle (88.9%) and lower rates of vacutainer use (38.9%) than nurses (13.3%, 53.3%, and 60.0%). Interns preferred to use safety devices (88.9%) rather than nurses (40.0%). The number of overall blood cultures and SD blood cultures was 9,053 and 555, respectively. While the overall blood volume of aerobic bottles was 2.6±2.1 mL, those of SD blood cultures were 5.0±3.0 mL in aerobic bottles and 6.0±3.0 mL in anaerobic bottles. Positive and contamination rates were 6.5% and 0.72% with SD blood cultures and 6.2% and 0.3% with overall blood cultures. Conclusions: The introduction of the safety device would encourage healthcare workers to collect adequate blood volume as well as lead to safer practices in the ER.

Keywords: blood culture, needlestick, safety device, volume

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Authors: Joshua N. Edokpayi, John O. Odiyo, Patience P. Shikwambana

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Water is a very rare natural resource in South Africa. Ga-Selati River is used for both domestic and industrial purposes. This study was carried out in order to assess the quality of Ga-Selati River in a mining area of Limpopo Province-Phalaborwa. The pH, Electrical Conductivity (EC) and Total Dissolved Solids (TDS) were determined using a Crinson multimeter while turbidity was measured using a Labcon Turbidimeter. The concentrations of Al, Ca, Cd, Cr, Fe, K, Mg, Mn, Na and Pb were analysed in triplicate using a Varian 520 flame atomic absorption spectrometer (AAS) supplied by PerkinElmer, after acid digestion with nitric acid in a fume cupboard. The average pH of the river from eight different sampling sites was 8.00 and 9.38 in wet and dry season respectively. Higher EC values were determined in the dry season (138.7 mS/m) than in the wet season (96.93 mS/m). Similarly, TDS values were higher in dry (929.29 mg/L) than in the wet season (640.72 mg/L) season. These values exceeded the recommended guideline of South Africa Department of Water Affairs and Forestry (DWAF) for domestic water use (70 mS/m) and that of the World Health Organization (WHO) (600 mS/m), respectively. Turbidity varied between 1.78-5.20 and 0.95-2.37 NTU in both wet and dry seasons. Total hardness of 312.50 mg/L and 297.75 mg/L as the concentration of CaCO3 was computed for the river in both the wet and the dry seasons and the river water was categorised as very hard. Mean concentration of the metals studied in both the wet and the dry seasons are: Na (94.06 mg/L and 196.3 mg/L), K (11.79 mg/L and 13.62 mg/L), Ca (45.60 mg/L and 41.30 mg/L), Mg (48.41 mg/L and 44.71 mg/L), Al (0.31 mg/L and 0.38 mg/L), Cd (0.01 mg/L and 0.01 mg/L), Cr (0.02 mg/L and 0.09 mg/L), Pb (0.05 mg/L and 0.06 mg/L), Mn (0.31 mg/L and 0.11 mg/L) and Fe (0.76 mg/L and 0.69 mg/L). Results from this study reveal that most of the metals were present in concentrations higher than the recommended guidelines of DWAF and WHO for domestic use and the protection of aquatic life.

Keywords: contamination, mining activities, surface water, trace metals

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Authors: Arfan Ali, Idrees Ahmad Nasir

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Potato (Solanum tuberosum) is an important cash crop and popular vegetable in Pakistan and throughout the world. Cold storage of potatoes accelerates the conversion of starch into reduced sugars (glucose and fructose). This process causes dry mass and bitter taste in the potatoes that are not acceptable to end consumers. In the current study, the phosphofructokinase B gene was cloned into the pET-30 vector for protein expression and the pCambia-1301 vector for plant expression. Amplification of a 930bp product from an E. coli strain determined the successful isolation of the phosphofructokinase B gene. Restriction digestion using NcoI and BglII along with the amplification of the 930bp product using gene specific primers confirmed the successful cloning of the PfkB gene in both vectors. The protein was expressed as a His-PfkB fusion protein. Western blot analysis confirmed the presence of the 35 Kda PfkB protein when hybridized with anti-His antibodies. The construct Fani-01 was evaluated transiently using a histochemical gus assay. The appearance of blue color in the agroinfiltrated area of potato leaves confirmed the successful expression of construct Fani-01. Further, the area displaying gus expression was evaluated for PfkB expression using ELISA. Moreover, PfkB gene expression evaluated through transient expression determined successful gene expression and highlighted its potential utilization for stable expression in potato to reduce sweetening due to long-term storage.

Keywords: potato, Solanum tuberosum, transformation, PfkB, anti-sweetening

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Authors: Abdul-Hakeem Olatunji Abiola, A. E. Adeniran, A. O. Ajimo, A. B. Lamilisa

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Introduction: Conventional wastewater treatment technology has been found to fail in developing countries because they are expensive to construct, operate and maintain. Constructed wetlands are nowadays considered as a low-cost alternative for effective wastewater treatment, especially where suitable land can be available. This study aims to evaluate the performance of the constructed wetland vegetated with water hyacinth (Eichhornia crassipes) plant for the treatment of wastewater. Methodology: The sub-surface flow wetland used for this study was an experimental scale constructed wetland consisting of four beds A, B, C, and D. Beds A, B, and D were vegetated while bed C which was used as a control was non-vegetated. This present study presents the results from bed B vegetated with water hyacinth (Eichhornia crassipes) and control bed C which was non-vegetated. The influent of the experimental scale wetland has been pre-treated with sedimentation, screening and anaerobic chamber before feeding into the experimental scale wetland. Results: pH and conductivity level were more reduced, colour of effluent was more improved, nitrate, iron, phosphate, and chromium were more removed, and dissolved oxygen was more improved in the water hyacinth bed than the control bed. While manganese, nickel, cyanuric acid, and copper were more removed from the control bed than the water hyacinth bed. Conclusion: The performance of the experimental scale constructed wetland bed planted with water hyacinth (Eichhornia crassipes) is better than that of the control bed. It is therefore recommended that plain bed without any plant should not be encouraged.

Keywords: constructed experimental scale wetland, domestic sewage, treatment, water hyacinth

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Authors: Debabrata Das, Preeti Mishra

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A two-phase anaerobic process combining biohydrogen followed by biomethane (biohythane technology) serves as an environment-friendly and economically sustainable approach for the improved valorization of organic wastes. Suitability of the pure cultures like Klebsiela pneumonia, C. freundii, B. coagulan, etc. and mixed acidogenic cultures for the biohydrogen production was already studied. The characteristics of organic wastes play a critical role in biohydrogen production. The choice of an appropriate combination of complementary organic wastes can vastly improve the bioenergy generation besides achieving the significant cost reduction. Suitability and economic viability of using the groundnut deoiled cake (GDOC), mustard deoiled cake (MDOC), distillers’ dried grain with soluble (DDGS) and algal biomass (AB) as a co-substrate were studied for a biohythane production. Results show that maximum gaseous energy of 20.7, 9.3, 16.7 and 15.6 % was recovered using GDOC, MDOC, DDGS and AB in the two stage biohythane production, respectively. Both GDOC and DDGS were found to be better co-substrates as compared to MDOC and AB in terms of hythane production, respectively. The maximum cumulative hydrogen and methane production of 150 and 64 mmol/L were achieved using GDOC. Further, 98 % reduction in substrate input cost (SIC) was achieved using the co-supplementation procedure.

Keywords: Biohythane, algal biomass, distillers’ dried grain with soluble (DDGS), groundnut deoiled cake (GDOC), mustard deoiled cake (MDOC)

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Authors: Abdulsalam. I. Rafida, Marwa. F. Elalem, Hasna. E. Alemam

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This study aimed at testing the various types of water in some areas of the city of Tripoli, Libya for the presence of Heterotrophic Aerobic Bacteria (HAB), and anaerobic Sulphate Reducing Bacteria (SRB). The water samples under investigation included rainwater accumulating on the ground, sewage water (from the city sewage treatment station, sulphate water from natural therapy swimming sites), and sea water (i.e. sea water exposed to pollution by untreated sewage water, and unpolluted sea water from specific locations). A total of 20 samples have been collected distributed as follows: rain water (8 samples), sewage water (6 samples), and sea water (6 samples). An up-to-date method for estimation has been used featuring readymade solutions i.e. (BARTTM test for HAB and BARTTM test for SRB). However, with the exception of one rain water sample, the results have indicated that the target bacteria have been present in all samples. Regarding HAB bacteria the samples have shown a maximum average of 7.0 x 106 cfu/ml featuring sewage and rain water and a minimum average of 1.8 x 104 cuf/ml featuring unpolluted sea water collected from a specific location. As for SRB bacteria; a maximum average of 7.0 x 105 cfu/ml has been shown by sewage and rain water and a minimum average of 1.8 x 104 cfu/ml by sewage and sea water. The above results highlight the relationship between pollution and the presence of bacteria in water particularly water collected from specific locations, and also the presence of bacteria as the result of the use of water provided that a suitable environment exists for its growth.

Keywords: heterotrophic aerobic bacteria (HAB), sulphate reducing bacteria (SRB), water, environmental sciences

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Authors: Fatma Y. Meligy

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Objectives: The objectives of this study aimed to isolate and characterize mesenchymal stem cells (MSCs) derived from synovial membrane. Then to assess the potentiality of myogenic differentiation of these isolated MSCs. Methods: The MSCs were isolated from synovial membrane by digestion method. Three adult rats were used. The 5 -azacytidine was added to the cultured cells for one day. The isolated cells and treated cells are assessed using immunoflouresence, flowcytometry, PCR and real time PCR. Results: The isolated stem cells showed morphological aspect of stem cells they showed strong positivity to CD44 and CD90 in immunoflouresence while in CD34 and CD45 showed negative reaction. The treated cells with 5-azacytidine was shown to have positive reaction for desmin. Flowcytometric analysis showed that synovial MSCs had strong positive percentage for CD44(%98)and CD90 (%97) and low percentage for CD34 & CD45 while the treated cells showed positive percentage for myogenic marker myogenin (85%). As regard the PCR and Real time PCR, the treated cells showed positive reaction to the desmin primer. Conclusion: The adult MSCs were isolated successfully from synovial membrane and characterized with stem cell markers. The isolated cells could be differentiated in vitro into myogenic cells. These differentiated cells could be used in auto-replacement of diseased or traumatized muscle cells as a regenerative therapy for muscle disorders and trauma.

Keywords: mesenchymal stem cells, synovial membrane, myogenic differentiation

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Authors: J. Herrero, D. Puigserver, I. Nijenhuis, K. Kuntze, J. M. Carmona

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In the transition zone between aquifers and basal aquitards, the perchloroethene (PCE)-pools are more recalcitrant than those elsewhere in the aquifer. Although biodegradation of chloroethenes occur in this zone, it is a slow process and a remediation strategy is needed. The aim of this study is to demonstrate that combined strategy of biostimulation and in situ chemical reduction (ISCR) is more efficient than the two separated strategies. Four different microcosm experiments with sediment and groundwater of a selected field site where an aged pool exists at the bottom of a transition zone were designed under i) natural conditions, ii) biostimulation with lactic acid, iii) ISCR with zero-value iron (ZVI) and under iv) a combined strategy with lactic acid and ZVI. Biotic and abiotic dehalogenation, terminal electron acceptor processes and evolution of microbial communities were determined for each experiment. The main results were: i) reductive dehalogenation of PCE-pools occurs under sulfate-reducing conditions; ii) biostimulation with lactic acid supports more pronounced reductive dehalogenation of PCE and trichloroethene (TCE), but results in an accumulation of 1,2-cis-dichloroethene (cDCE); iii) ISCR with ZVI produces a sustained dehalogenation of PCE and its metabolites iv) combined strategy of biostimulation and ISCR results in a fast dehalogenation of PCE and TCE and a sustained dehalogenation of cisDCE. These findings suggest that biostimulation and ISCR with ZVI are the most suitable strategies for a complete reductive dehalogenation of PCE-pools in the transition zone and further to enable the dissolution of dense non-aqueous phase liquids.

Keywords: aged PCE-pool, anaerobic microcosm experiment, biostimulation, in situ chemical reduction, natural attenuation

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Authors: Preethi Grace Theva Neethi Dhas

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A healthy human gut microbiome has commensal and symbiotic functions in digestion and is a decisive factor for human health. The soil microbiome is a crucial component in the ecosystem of soils and their health and resilience. Changes in soil microbiome are linked to human health. Ever since the industrial era, the human and the soil microbiome have been going through drastic changes. The soil microbiome has changed due to industrialization and extensive agricultural practices, whereas humans have less contact with soil and increased intake of highly processed foods, leading to changes in the human gut microbiome. Regenerating the soil becomes crucial in maintaining a healthy ecosystem. The nutrients, once obtained from the soil, need to be given back to the soil. Soil degradation needs to be addressed in effective ways, like adding organic nutrients back to the soil. Manure from animals and humans needs to be returned to the soil, which can complete the nutrient cycle in the soil. On the other hand, fecal sludge management (FSM) is a growing concern in many parts of the developing world. Hence, it becomes crucial to treat and reuse fecal sludge in a safe manner, i.e., low in risk to human health. Co-composting fecal sludge with organic wastes is a practice that allows the safe management of fecal sludge and the safe application of nutrients to the soil. This paper will discuss the possible impact of co-composting fecal sludge with coconut choir waste on the soil, water, and ecosystem at large. Impact parameters like nitrogen, phosphorus, and fecal coliforms will be analyzed. The overall impact of fecal sludge application on the soil will be researched and presented in this study.

Keywords: fecal sludge management, nutrient cycle, soil health, composting

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Authors: Aprajita Singh, Anamika Tripathi, Surya P. Dwivedi

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Moradabad is a small town in the Northern area of Uttar Pradesh, India. It is situated on the bank of river Ramganga which is also known as ‘Brass City of India’. There is eventually increase in the environmental pollution due to uncontrolled and inappropriate e-waste burning (recycling) activities which have been reported in many areas of Moradabad. In this paper, analysis of toxic heavy metals, causing pollution to the surrounding environment released from the e-waste burning and much other recycling process. All major e-waste burning sites are situated on the banks of the river which is burned in open environmental conditions. Soil samples were collected from seven (n=3) different sites including control site, after digestion of soil samples using triacid mixture, analysis of different toxic metals (Pb, Ar, Hg, Cd, Cr, Cu, Zn, Fe, and Ni) has been carried out with the help of instrument ICP-AAS. After the study, the outcome is that the soil of those areas contains a relatively high level of the toxic metals in order of Cu>Fe>Pb>Cd>Cr>Zn>Ar>Hg. The concentration of Cd, Pb, Cr, Ar and Zn (the majority of samples experimentally proved) exceeded the maximum standard level of WHO. Sequentially this study showed that uncontrolled e-waste processing operations caused serious pollution to local soil and release of toxic metals in the environment is also causing adverse effect on the health of people living in the nearby areas making them more prone to various harmful diseases.

Keywords: brass city, environment pollution, e-waste, toxic heavy metals

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Authors: Cora Fernandez Dacosta, John A. Posada, Andrea Ramirez

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The biodegradable family of polymers polyhydroxyalkanoates are interesting substitutes for convectional fossil-based plastics. However, the manufacturing and environmental impacts associated with their production via intracellular bacterial fermentation are strongly dependent on the raw material used and on energy consumption during the extraction process, limiting their potential for commercialization. Industrial wastewater is studied in this paper as a promising alternative feedstock for waste valorization. Based on results from laboratory and pilot-scale experiments, a conceptual process design, techno-economic analysis and life cycle assessment are developed for the large-scale production of the most common type of polyhydroxyalkanoate, polyhydroxbutyrate. Intracellular polyhydroxybutyrate is obtained via fermentation of microbial community present in industrial wastewater and the downstream processing is based on chemical digestion with surfactant and hypochlorite. The economic potential and environmental performance results help identifying bottlenecks and best opportunities to scale-up the process prior to industrial implementation. The outcome of this research indicates that the fermentation of wastewater towards PHB presents advantages compared to traditional PHAs production from sugars because the null environmental burdens and financial costs of the raw material in the bioplastic production process. Nevertheless, process optimization is still required to compete with the petrochemicals counterparts.

Keywords: circular economy, life cycle assessment, polyhydroxyalkanoates, waste valorization

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Authors: Amtul Jamil Sami

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Apis mellifera is an insect of immense economic importance lives on rich carbohydrate diet including cellulose, nectar, honey and pollen. The carbohydrate metabolism in A mellifera has not been understood fully, as there are no data available, on the functional expression of cellulase gene. The cellulose hydrolyzing enzyme is required for the digestion of pollen cellulose wall, to release the important nutrients (amino acids, minerals, vitamins etc.) from the pollen. A dissection of Apis genome had revealed that there is one gene present for the expression of endo-beta-1,4-glucanase, for cellulose hydrolysis. In the presented work, functional expression of endo-beta-1,4 glucanase gene is reported. Total soluble proteins of the honey bee were isolated and were tested cellulose hydrolyzing enzyme activity, using carboxy-methyl cellulose, as a substrate. A mellifera proteins were able to hydrolyze carboxy-methyl cellulose, confirming its endo- type mode of action. Endo beta-1,4 glucanase enzyme was only present in the gut tissues, no activity was detected in the salivary glands. The pH optima of the enzyme were in the acidic pH range of 4-5-5-0, indicating its metabolic role in the acidic stomach of A mellifera. The reported enzyme is unique, as endo-beta- 1,4 glucanase was able to generate non reducing sugar, as an end product. The results presented, are supportive to the information that the honey bee is capable of producing its novel endo-beta-1,4 glucanase. Further it could be helpful, in understanding, the carbohydrate metabolism in A mellifera.

Keywords: honey bees, Endo-beta 1, 4- glucanase, Apis mellifera, functional expression

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Authors: Vibha Sinha, Sumedha Chakma

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Arsenic (As) contamination in drinking water is a serious concern worldwide resulting in severe health maladies. To tackle this problem, several hydrogel based matrix which selectively uptake toxic metals from contaminated water has increasingly been examined as a potential practical method for metal removal. The major concern in hydrogels is low stability of matrix, resulting in poor performance. In this study, the potential of hybrid hydrogel-biochar matrix synthesized from natural plant polymers, specific for As removal was explored. Various compositional and functional group changes of the elements contained in the matrix due to the adsorption of As were identified. Moreover, to resolve the stability issue in hydrogel matrix, optimum and effective mixing of hydrogel with biochar was studied. Mixing varied proportions of matrix components at the time of digestion process was tested. Preliminary results suggest that partial premixing methods may increase the stability and reduce cost. Addition of nanoparticles and specific catalysts with different concentrations of As(III) and As(V) under batch conditions was performed to study their role in performance enhancement of the hydrogel matrix. Further, effect of process parameters, optimal uptake conditions and detailed mechanism derived from experimental studies were suitably conducted. This study provides an efficient, specific and a low-cost As removal method that offers excellent regeneration abilities which can be reused for value.

Keywords: arsenic, catalysts, hybrid hydrogel-biochar, water purification

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Authors: N. Anusree, P. Sughosh, G. L. Sivakumar Babu

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Municipalities throughout the world are marred with serious issues related to the Municipal Solid Waste (MSW) collection, treatment, and safe disposal. While the Waste Management sector contributes around 3-9 % of the overall anthropogenic methane emission, measures towards mitigating these emissions are rarely given attention in developing countries. In the case of Bangalore, India, around 5680 tons of MSW is generated in a day, and its collection and treatment efficiency are around 90-95 % and 26.4 %, respectively. About 33.4 % of the waste collected is directly landfilled without any treatment, further aggravating the situation. The potential of reducing the emissions emanating from the MSW of Bangalore city without any severe consequences on the current MSW management practices is evaluated in this study. Three emission scenarios consisting of the baseline condition (current practices – Case-1), the application of biocovers for methane oxidation in the dumpsites (case-2), and the diversion of Organic Fraction of MSW (OFMSW) along with the application of biocovers (case-3) are evaluated and compared with each other. The emissions are calculated based on the aerobic and anaerobic stochiometric relations for the three scenarios. Laboratory scale column studies are carried out to determine the methane oxidation potential of three different biocover material (digested MBT (mechanically biologically treated) waste, Fresh MBT waste, and charcoal amended with fresh MBT waste). The results shown that around 40 % and 83 % reduction in carbon emissions can be achieved in case 3 and 2 in comparison to the baseline condition. The study clearly shows that with minor changes in the waste management practices, substantial reductions in the carbon emissions can be attained in Bangalore City.

Keywords: MSW, biocover, composting, carbon emission

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Authors: Kaniz Roksana, Aluthgun Hewage Shaini, Cheng Zhu

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Lead is environmentally hazardous because it may persist for a long time in soil, water, and air, and it can travel large distances when carried by wind or water. Lead is toxic to many different species of organisms and has the potential to disrupt ecosystem stability. Moreover, lead can contaminate crops and livestock, which can then have an adverse effect on human health. This study was conducted to use the enzyme-induced calcium carbonate precipitation (EICP) technique from soybean crude extract urease along coconut fiber derived biochar’s (CFB) to bioremediate lead. To study the desorption rates of heavy metals from the soil, lead (Pb) was added to the soil at load ratios of 50 and 100 mg/kg. There were five separate treatment soil columns created: control sample, only CFB, only EICP, EICP with 2% (w/w) CFB, and EICP with 4% (w/w) CFB. Laboratory scale experiment demonstrates significant lead removal from soil. The amount of CaCO₃ precipitated in the soil was measured using a gravimetric acid digestion test, which related heavy metal desorption to the amount of precipitated calcium carbonate. These findings were validated using a scanning electron microscope (SEM), which revealed calcium carbonate and lead coprecipitation. As a result, the study reveals that the EICP technique, in conjunction with coconut fiber biochar, could be an efficient alternative in the remediation of heavy metal ion-contaminated soils.

Keywords: enzyme induced calcium carbonate precipitation (EICP), coconut fiber derived biochar’s (CFB), bioremediation, heavy metal

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