Pooja Shivanand - Academia.edu (original) (raw)
Papers by Pooja Shivanand
Malaysian journal of microbiology, Apr 1, 2024
The Philippine journal of science, Sep 20, 2023
Biosensors and Bioelectronics, Feb 29, 2024
Proceedings of the National Academy of Sciences, India, Section B: biological sciences, Jul 16, 2019
Background: Due to the effect of midgut bacteria on proliferation of parasites and their potentia... more Background: Due to the effect of midgut bacteria on proliferation of parasites and their potential as paratransgenesis tools, their identification in malaria vector mosquitoes is important. Anopheles culicifacies s.l. is one of the main malaria vectors in Asia; however, its midgut microbiota remains un-studied. This work was primarily designed to isolate potential candidates for use in a paratransgenesis approach, but also to give a picture of the midgut microbiota of wild-caught An. culicifacies larvae and adults from the southeast corner of Iran, which has the highest malaria endemicity in the country. Methods: A total of 68 larvae and 34 adult females (newly eclosed and older) from three different biotopes in Iran were analyzed for their midgut microflora. The mosquitoes had their midgut bacterial contents plated on three different culture media (brain heart agar, nutrient agar and blood agar) yielding 57 bacterial isolates. The 16S rRNA genes of the isolates were sequence analyzed for species designation, which then was confirmed by biochemical analysis.
Brazilian Journal of Microbiology
Journal of Food Science and Technology, 2021
In recent years, rapid detection methods such as polymerase chain reaction (PCR) and quantitative... more In recent years, rapid detection methods such as polymerase chain reaction (PCR) and quantitative real-time PCR (qPCR) have been continuously developed to improve the detection of food-borne pathogens in food samples. The recent developments of PCR and qPCR in the detection and identification of these food-borne pathogens are described and elaborated throughout this review. Specifically, further developments and improvements of qPCR are discussed in detecting Salmonella and norovirus. Promising advances in these molecular detection methods have been widely used to prevent human food-borne illnesses and death caused by the food-borne pathogens. In addition, this review presents the limitations and challenges of the detection methods which include conventional culture method and conventional PCR method in detecting Salmonella and norovirus. Furthermore, several advances of qPCR such as viability PCR (vPCR) and digital PCR (dPCR) have been discussed in the detection of Salmonella and norovirus. Good practice of analysis of the food-borne pathogens and other contaminants in the food industry as well as the advancement of molecular detection methods will help improve and ensure food safety and food quality.
Annals of Microbiology, 2012
A newly isolated biopolymer-degrading halophilic bacterium, Halomonas sp. strain PS47, yielded hi... more A newly isolated biopolymer-degrading halophilic bacterium, Halomonas sp. strain PS47, yielded higher cellulase activity (0.0076 U/ml) in mineral salt medium (MM63). Activity increased to 0.029 U/ml when carboxymethyl cellulose (0.5 % w/v) was used as carbon source and further to 0.138 U/ml when a combination of yeast extract and peptone was used as nitrogen source. Enzyme secretion was maximal during late exponential and stationary phases (0.15 U/ml, 48 h). Among different agro-residues (1 % w/v), wheat bran gave the highest activity (0.12 U/ml) at pH 7.5, 30°C and 6 % (w/v) NaCl. The cellulase exhibited higher activity at pH 7.1 and 50°C. The enzyme exhibited activity over a wide range of NaCl concentrations (0-4 M). Optimum activity was at 0-1 M NaCl. At 4 M NaCl, activity was reduced to 65 % of the initial value. The present investigation thus contributes to the limited information available on halostable cellulases.
Journal of applied biology and biotechnology, 2023
Molecules, Oct 3, 2022
This article is an open access article distributed under the terms and conditions of the Creative... more This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY
Biodiversitas Journal of Biological Diversity
Microbes have gained much attention for their application in the bioremediation of petroleum cont... more Microbes have gained much attention for their application in the bioremediation of petroleum contaminated areas, and biofilm producing bacteria are considered potential candidates for this purpose. The aim of this study was to screen six oil degrading bacterial strains for their ability to form biofilm, and to measure the crude oil degradation efficiency of selected strains. It was found that only two strains, Micrococcus sp. UBDBH15 and Pseudomonas sp. UBDBH26 were categorized as strong biofilm producers, with the latter showing the highest amount of biofilm formed. Under the conditions tested, Pseudomonas sp. UBDBH26 was also found to have the highest degradation rate (0.0047 g/day) with a significant degradation of 7.07 ± 3.02 and 11.38 ± 2.93 % of crude oil after 7 and 14 days respectively. Micrococcus sp. UBDBH15 had a lower rate of degradation (0.0033 g/day), resulting in a non-significant degradation. However, in comparison with a non-biofilm producer, Enterobacter sp. UBDBH06, this study suggested that biofilm might enhance the degradation of oil, but further studies are needed to confirm this assumption.
Molecules
Agarwood, popularly known as oudh or gaharu, is a fragrant resinous wood of high commercial value... more Agarwood, popularly known as oudh or gaharu, is a fragrant resinous wood of high commercial value, traded worldwide and primarily used for its distinctive fragrance in incense, perfumes, and medicine. This fragrant wood is created when Aquilaria trees are wounded and infected by fungi, producing resin as a defense mechanism. The depletion of natural agarwood caused by overharvesting amidst increasing demand has caused this fragrant defensive resin of endangered Aquilaria to become a rare and valuable commodity. Given that instances of natural infection are quite low, artificial induction, including biological inoculation, is being conducted to induce agarwood formation. A long-term investigation could unravel insights contributing toward Aquilaria being sustainably cultivated. This review will look at the different methods of induction, including physical, chemical, and biological, and compare the production, yield, and quality of such treatments with naturally formed agarwood. Phar...
Journal of Tropical Biodiversity and Biotechnology
Conventional products have a role in addressing the thriving universal demands for biologically a... more Conventional products have a role in addressing the thriving universal demands for biologically active substances. Since the South China Sea is a prodigious province of geostrategic and mercantile importance, it meets the basic needs of people who dwell there. The South China Sea is dominant in mangrove biodiversity which, represents 11.4% of the world’s 15.5 million hectares of mangrove forest. Mangroves are harbored by multifaceted fungal communities that represent the second colossal ecological breed of marine fungi. The symbiotic association between the plants and fungi stimulates the bioactive components such as alkaloid, depsipeptides, cyclic peptides, quinone, terpenes, lactones, terpenoid, flavonoid, phenolic acid, steroids. These components have multifaceted pharmacological activities likely, anti-inflammatory, antidiabetic, anticancer, antioxidant, and antimicrobial. This review article attempts to present a piece of insightful information currently being explored on the b...
Journal of Environmental Science and Health, Part A, 2020
Microbes that can be cultured and degrade petroleum are of particular interest for biotechnology ... more Microbes that can be cultured and degrade petroleum are of particular interest for biotechnology such as bioremediation. This study aims to isolate and identify culturable petroleum-degrading bacteria and fungi from Brunei Darussalam, which has not previously been explored. A total of eight bacterial and nine fungal isolates that could degrade petroleum were obtained from petroleum-contaminated water or soil samples. DNA barcoding using 16S rRNA gene sequence identified five different bacterial genera which were Bacillus, Enterobacter, Micrococcus, Pseudoaltermonas and Pseudomonas. DNA barcoding using rRNA-ITS gene sequence identified nine different fungal taxa which were Aspergillus, Cladosporium, Exophiala, Flavodon, Hypocreales, Nectriaceae, Penicillium, Peniophora and Trichoderma. Biolog provided additional support to the identification of some isolates. This study is the first to report these unique microbes from Brunei Darussalam, which are of ecological and biotechnological value.
Rhizomicrobiome Dynamics in Bioremediation, 2021
Biodiversitas Journal of Biological Diversity, 2021
Microbial diversity in Brunei Darussalam located on northwest coast of Borneo is not well explore... more Microbial diversity in Brunei Darussalam located on northwest coast of Borneo is not well explored. This study aimed to isolate and identify culturable marine fungi and bacteria from coastal regions. Microbial identification was carried out via DNA barcoding using rRNA-ITS marker for fungi and 16S rRNA marker for bacteria. Nine marine fungal isolates were identified to five genera as Aspergillus, Penicillium, Phialemoniopsis, Purpureocillium and Trametes. However, only one genus, Pseudoalteromonas was identified from ten marine bacterial isolates with at least three different Pseudoalteromonas species expected from the isolation. This study provided an insight into the diversity of culturable marine microbes from a coastal ecosystem. As the microbes were found culturable and can easily grow under saline conditions, they are potential of considerable biotechnological interest.
Biodiversitas Journal of Biological Diversity, 2021
Aquilaria is an important agarwood-producing tree, but naturally occurring agarwood is rare. Arti... more Aquilaria is an important agarwood-producing tree, but naturally occurring agarwood is rare. Artificial induction method is the most commonly used to produce agarwood in a short time. Out of the 21 species of Aquilaria, Brunei Darussalam houses two species that produce high quality agarwood, namely Aquilaria beccariana and A. microcarpa. The aims of this study are to artificially induce agarwood formation using eight different induction treatments: six fungal inoculant treatments with two control treatments, and to isolate and identify fungal strains from naturally infected agarwood tree using DNA barcoding. Agarwood formation was observed at four different time intervals i.e. 1, 3, 6 and 9 months of post-inoculation. Results exhibited that the area and the total length of discoloration zone were significantly greater at 6 and 9 months of incubation period. Induction treatment did not significantly affect the area and the total length of discoloration. FTIR analysis revealed that two aromatic compounds (C-H and CC stretching) were detected at the wavelength ranges of 3000-2800 cm-1 , and 1500-1400 cm-1 respectively, and OH group was detected at 3600-3400 cm-1. Nine fungal strains from Aspergillus, Fusarium, Mucor and Trichosporon were isolated and identified from a naturally infected agarwood, and the last two fungi (Mucor and Trichosporon) were never been previously reported.
Current Science, 2019
Fungi are the most diverse and adaptable group of living organisms. The literature highlights tha... more Fungi are the most diverse and adaptable group of living organisms. The literature highlights that colonization density and fungal diversity can be affected by factors such as salinity, pH, temperature, altitude and availability of nutrients. The present article discusses isolation and characterization of fungi from different ecosystems like forests, mangroves and coastal areas. Fungal interaction with ecosystems is reported to play an important role in the biodiversity, survivability, propagation and productivity of plants. Fungi have established a beneficial association with numerous hosts and have the capability to degrade environmental pollutants such as hydrocarbons and by-products. Hence, fungi and fungal products like extracellular hydrolytic and lignin-degrading enzymes, and surfactants offer eco-friendly and cost-effective strategies to address pollution, especially with respect to bioremediation of oil spills and polyaromatic hydrocarbons.
Surfactants are amphiphatic compounds containing both hydrophilic and hydrophobic groups, which a... more Surfactants are amphiphatic compounds containing both hydrophilic and hydrophobic groups, which are capable of lowering surface or interfacial tension. Considering the advantages of using biosurfactants produced by microorganisms, the aim of this study was to develop and characterise a biosurfactant produced by Trichoderma sp. MK116452 isolated from Seri Chermin filling station, Brunei Darussalam, and to examine its potential application in microbial enhanced oil recovery (MEOR). The microorganism was cultured in a minimal salt medium containing 1% crude oil as a carbon source and a cell-free culture broth was used to screen its efficiency in producing biosurfactants. Characterisation of the biosurfactant showed the presence of glycosides and fatty acids, suggesting its glycolipidic nature. The isolated biosurfactant showed no toxicity to the micro-crustacean Artemia salina or to red bean (Vigna angularis). The extracted biosurfactant was effective at recovering up to 60% of crude o...
Scientia Bruneiana, 2019
Domestic and municipal solid wastes pose environmental concerns and health risks, suggesting the ... more Domestic and municipal solid wastes pose environmental concerns and health risks, suggesting the importance of reduced waste disposal. Food waste accounts for the highest percentage of solid waste in Brunei. An effective strategy is to convert food waste into beneficial products such as enzymes and biofertilizers. This study reports utilization of inedible fruit peels as substrates for enzyme production. A fermentation medium prepared by adding fruit peels, brown sugar and water was allowed to ferment over a period of time. Total carbohydrate content was found to be highest in the fermented orange sample (37.87 ± 4.7 mg/mL) followed by pineapple and banana samples, 11.98 ± 1.45 mg/mL and 10.60 ± 0.45 mg/mL, respectively. Pineapple sample showed the highest concentration of reducing sugar (11.93 mg/mL at week 2 and 3.31 mg/mL after 3 months). Enzyme assay showed that citrus fruits like oranges, yield high activities of enzymes like protease (0.129 U/mL), α-amylase (7.261 ± 0.83 U/mL)...
Malaysian journal of microbiology, Apr 1, 2024
The Philippine journal of science, Sep 20, 2023
Biosensors and Bioelectronics, Feb 29, 2024
Proceedings of the National Academy of Sciences, India, Section B: biological sciences, Jul 16, 2019
Background: Due to the effect of midgut bacteria on proliferation of parasites and their potentia... more Background: Due to the effect of midgut bacteria on proliferation of parasites and their potential as paratransgenesis tools, their identification in malaria vector mosquitoes is important. Anopheles culicifacies s.l. is one of the main malaria vectors in Asia; however, its midgut microbiota remains un-studied. This work was primarily designed to isolate potential candidates for use in a paratransgenesis approach, but also to give a picture of the midgut microbiota of wild-caught An. culicifacies larvae and adults from the southeast corner of Iran, which has the highest malaria endemicity in the country. Methods: A total of 68 larvae and 34 adult females (newly eclosed and older) from three different biotopes in Iran were analyzed for their midgut microflora. The mosquitoes had their midgut bacterial contents plated on three different culture media (brain heart agar, nutrient agar and blood agar) yielding 57 bacterial isolates. The 16S rRNA genes of the isolates were sequence analyzed for species designation, which then was confirmed by biochemical analysis.
Brazilian Journal of Microbiology
Journal of Food Science and Technology, 2021
In recent years, rapid detection methods such as polymerase chain reaction (PCR) and quantitative... more In recent years, rapid detection methods such as polymerase chain reaction (PCR) and quantitative real-time PCR (qPCR) have been continuously developed to improve the detection of food-borne pathogens in food samples. The recent developments of PCR and qPCR in the detection and identification of these food-borne pathogens are described and elaborated throughout this review. Specifically, further developments and improvements of qPCR are discussed in detecting Salmonella and norovirus. Promising advances in these molecular detection methods have been widely used to prevent human food-borne illnesses and death caused by the food-borne pathogens. In addition, this review presents the limitations and challenges of the detection methods which include conventional culture method and conventional PCR method in detecting Salmonella and norovirus. Furthermore, several advances of qPCR such as viability PCR (vPCR) and digital PCR (dPCR) have been discussed in the detection of Salmonella and norovirus. Good practice of analysis of the food-borne pathogens and other contaminants in the food industry as well as the advancement of molecular detection methods will help improve and ensure food safety and food quality.
Annals of Microbiology, 2012
A newly isolated biopolymer-degrading halophilic bacterium, Halomonas sp. strain PS47, yielded hi... more A newly isolated biopolymer-degrading halophilic bacterium, Halomonas sp. strain PS47, yielded higher cellulase activity (0.0076 U/ml) in mineral salt medium (MM63). Activity increased to 0.029 U/ml when carboxymethyl cellulose (0.5 % w/v) was used as carbon source and further to 0.138 U/ml when a combination of yeast extract and peptone was used as nitrogen source. Enzyme secretion was maximal during late exponential and stationary phases (0.15 U/ml, 48 h). Among different agro-residues (1 % w/v), wheat bran gave the highest activity (0.12 U/ml) at pH 7.5, 30°C and 6 % (w/v) NaCl. The cellulase exhibited higher activity at pH 7.1 and 50°C. The enzyme exhibited activity over a wide range of NaCl concentrations (0-4 M). Optimum activity was at 0-1 M NaCl. At 4 M NaCl, activity was reduced to 65 % of the initial value. The present investigation thus contributes to the limited information available on halostable cellulases.
Journal of applied biology and biotechnology, 2023
Molecules, Oct 3, 2022
This article is an open access article distributed under the terms and conditions of the Creative... more This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY
Biodiversitas Journal of Biological Diversity
Microbes have gained much attention for their application in the bioremediation of petroleum cont... more Microbes have gained much attention for their application in the bioremediation of petroleum contaminated areas, and biofilm producing bacteria are considered potential candidates for this purpose. The aim of this study was to screen six oil degrading bacterial strains for their ability to form biofilm, and to measure the crude oil degradation efficiency of selected strains. It was found that only two strains, Micrococcus sp. UBDBH15 and Pseudomonas sp. UBDBH26 were categorized as strong biofilm producers, with the latter showing the highest amount of biofilm formed. Under the conditions tested, Pseudomonas sp. UBDBH26 was also found to have the highest degradation rate (0.0047 g/day) with a significant degradation of 7.07 ± 3.02 and 11.38 ± 2.93 % of crude oil after 7 and 14 days respectively. Micrococcus sp. UBDBH15 had a lower rate of degradation (0.0033 g/day), resulting in a non-significant degradation. However, in comparison with a non-biofilm producer, Enterobacter sp. UBDBH06, this study suggested that biofilm might enhance the degradation of oil, but further studies are needed to confirm this assumption.
Molecules
Agarwood, popularly known as oudh or gaharu, is a fragrant resinous wood of high commercial value... more Agarwood, popularly known as oudh or gaharu, is a fragrant resinous wood of high commercial value, traded worldwide and primarily used for its distinctive fragrance in incense, perfumes, and medicine. This fragrant wood is created when Aquilaria trees are wounded and infected by fungi, producing resin as a defense mechanism. The depletion of natural agarwood caused by overharvesting amidst increasing demand has caused this fragrant defensive resin of endangered Aquilaria to become a rare and valuable commodity. Given that instances of natural infection are quite low, artificial induction, including biological inoculation, is being conducted to induce agarwood formation. A long-term investigation could unravel insights contributing toward Aquilaria being sustainably cultivated. This review will look at the different methods of induction, including physical, chemical, and biological, and compare the production, yield, and quality of such treatments with naturally formed agarwood. Phar...
Journal of Tropical Biodiversity and Biotechnology
Conventional products have a role in addressing the thriving universal demands for biologically a... more Conventional products have a role in addressing the thriving universal demands for biologically active substances. Since the South China Sea is a prodigious province of geostrategic and mercantile importance, it meets the basic needs of people who dwell there. The South China Sea is dominant in mangrove biodiversity which, represents 11.4% of the world’s 15.5 million hectares of mangrove forest. Mangroves are harbored by multifaceted fungal communities that represent the second colossal ecological breed of marine fungi. The symbiotic association between the plants and fungi stimulates the bioactive components such as alkaloid, depsipeptides, cyclic peptides, quinone, terpenes, lactones, terpenoid, flavonoid, phenolic acid, steroids. These components have multifaceted pharmacological activities likely, anti-inflammatory, antidiabetic, anticancer, antioxidant, and antimicrobial. This review article attempts to present a piece of insightful information currently being explored on the b...
Journal of Environmental Science and Health, Part A, 2020
Microbes that can be cultured and degrade petroleum are of particular interest for biotechnology ... more Microbes that can be cultured and degrade petroleum are of particular interest for biotechnology such as bioremediation. This study aims to isolate and identify culturable petroleum-degrading bacteria and fungi from Brunei Darussalam, which has not previously been explored. A total of eight bacterial and nine fungal isolates that could degrade petroleum were obtained from petroleum-contaminated water or soil samples. DNA barcoding using 16S rRNA gene sequence identified five different bacterial genera which were Bacillus, Enterobacter, Micrococcus, Pseudoaltermonas and Pseudomonas. DNA barcoding using rRNA-ITS gene sequence identified nine different fungal taxa which were Aspergillus, Cladosporium, Exophiala, Flavodon, Hypocreales, Nectriaceae, Penicillium, Peniophora and Trichoderma. Biolog provided additional support to the identification of some isolates. This study is the first to report these unique microbes from Brunei Darussalam, which are of ecological and biotechnological value.
Rhizomicrobiome Dynamics in Bioremediation, 2021
Biodiversitas Journal of Biological Diversity, 2021
Microbial diversity in Brunei Darussalam located on northwest coast of Borneo is not well explore... more Microbial diversity in Brunei Darussalam located on northwest coast of Borneo is not well explored. This study aimed to isolate and identify culturable marine fungi and bacteria from coastal regions. Microbial identification was carried out via DNA barcoding using rRNA-ITS marker for fungi and 16S rRNA marker for bacteria. Nine marine fungal isolates were identified to five genera as Aspergillus, Penicillium, Phialemoniopsis, Purpureocillium and Trametes. However, only one genus, Pseudoalteromonas was identified from ten marine bacterial isolates with at least three different Pseudoalteromonas species expected from the isolation. This study provided an insight into the diversity of culturable marine microbes from a coastal ecosystem. As the microbes were found culturable and can easily grow under saline conditions, they are potential of considerable biotechnological interest.
Biodiversitas Journal of Biological Diversity, 2021
Aquilaria is an important agarwood-producing tree, but naturally occurring agarwood is rare. Arti... more Aquilaria is an important agarwood-producing tree, but naturally occurring agarwood is rare. Artificial induction method is the most commonly used to produce agarwood in a short time. Out of the 21 species of Aquilaria, Brunei Darussalam houses two species that produce high quality agarwood, namely Aquilaria beccariana and A. microcarpa. The aims of this study are to artificially induce agarwood formation using eight different induction treatments: six fungal inoculant treatments with two control treatments, and to isolate and identify fungal strains from naturally infected agarwood tree using DNA barcoding. Agarwood formation was observed at four different time intervals i.e. 1, 3, 6 and 9 months of post-inoculation. Results exhibited that the area and the total length of discoloration zone were significantly greater at 6 and 9 months of incubation period. Induction treatment did not significantly affect the area and the total length of discoloration. FTIR analysis revealed that two aromatic compounds (C-H and CC stretching) were detected at the wavelength ranges of 3000-2800 cm-1 , and 1500-1400 cm-1 respectively, and OH group was detected at 3600-3400 cm-1. Nine fungal strains from Aspergillus, Fusarium, Mucor and Trichosporon were isolated and identified from a naturally infected agarwood, and the last two fungi (Mucor and Trichosporon) were never been previously reported.
Current Science, 2019
Fungi are the most diverse and adaptable group of living organisms. The literature highlights tha... more Fungi are the most diverse and adaptable group of living organisms. The literature highlights that colonization density and fungal diversity can be affected by factors such as salinity, pH, temperature, altitude and availability of nutrients. The present article discusses isolation and characterization of fungi from different ecosystems like forests, mangroves and coastal areas. Fungal interaction with ecosystems is reported to play an important role in the biodiversity, survivability, propagation and productivity of plants. Fungi have established a beneficial association with numerous hosts and have the capability to degrade environmental pollutants such as hydrocarbons and by-products. Hence, fungi and fungal products like extracellular hydrolytic and lignin-degrading enzymes, and surfactants offer eco-friendly and cost-effective strategies to address pollution, especially with respect to bioremediation of oil spills and polyaromatic hydrocarbons.
Surfactants are amphiphatic compounds containing both hydrophilic and hydrophobic groups, which a... more Surfactants are amphiphatic compounds containing both hydrophilic and hydrophobic groups, which are capable of lowering surface or interfacial tension. Considering the advantages of using biosurfactants produced by microorganisms, the aim of this study was to develop and characterise a biosurfactant produced by Trichoderma sp. MK116452 isolated from Seri Chermin filling station, Brunei Darussalam, and to examine its potential application in microbial enhanced oil recovery (MEOR). The microorganism was cultured in a minimal salt medium containing 1% crude oil as a carbon source and a cell-free culture broth was used to screen its efficiency in producing biosurfactants. Characterisation of the biosurfactant showed the presence of glycosides and fatty acids, suggesting its glycolipidic nature. The isolated biosurfactant showed no toxicity to the micro-crustacean Artemia salina or to red bean (Vigna angularis). The extracted biosurfactant was effective at recovering up to 60% of crude o...
Scientia Bruneiana, 2019
Domestic and municipal solid wastes pose environmental concerns and health risks, suggesting the ... more Domestic and municipal solid wastes pose environmental concerns and health risks, suggesting the importance of reduced waste disposal. Food waste accounts for the highest percentage of solid waste in Brunei. An effective strategy is to convert food waste into beneficial products such as enzymes and biofertilizers. This study reports utilization of inedible fruit peels as substrates for enzyme production. A fermentation medium prepared by adding fruit peels, brown sugar and water was allowed to ferment over a period of time. Total carbohydrate content was found to be highest in the fermented orange sample (37.87 ± 4.7 mg/mL) followed by pineapple and banana samples, 11.98 ± 1.45 mg/mL and 10.60 ± 0.45 mg/mL, respectively. Pineapple sample showed the highest concentration of reducing sugar (11.93 mg/mL at week 2 and 3.31 mg/mL after 3 months). Enzyme assay showed that citrus fruits like oranges, yield high activities of enzymes like protease (0.129 U/mL), α-amylase (7.261 ± 0.83 U/mL)...