Asian Journal of Research in Biochemistry

Optimization of Amylases Production from Fonio Using Central Composite Design

Adenike Oluwaseun, ADEFILA — 2025-08-28

This study aimed to optimize the production of amylases from grains of fonio which are of two main types; Digitaria exilis popular referred to as white fonio or Acha and Digitaria iburua, commonly known as black fonio or Iburu. These are with a view of determining the suitability of amylases from these grains as a suitable alternative to amylases from other tropical cereals. Whole grains of white and black fonio (100 g) were steeped in water and water containing phosphate salt for 24 h at room temperature and germinated in a locally constructed malting chamber for six (6) days. The most appropriate conditions of pH and temperature for the induction of amylases in these grains were selected based on central composite design (CCD). Amylase activities of crude extracts from harvested malts of white and black fonio were measured by assaying the reducing sugar released from starch using dinitrosalicylic acid method and soluble proteins were quantified by Bradford method. The induced amylases are a function of the days of germination as the highest amylase activity was obtained on second day (48 h) of germination with 17, 498 U/mg protein and 24, 337 U/mg protein for white and black fonio respectively. CCD suggested a cubic and quadratic model with range of values factor A (pH) and B (temperature) as limiting factors because any variation in themwill greatly affect the induced amylases which is the response. Specific activity (response) for both white and black fonio grains increases with an increase in both pH and temperature until further increase in these two factors resulted in a decrease in their responses (specific activity). CCD indicated that pH 6.5 and 22.5 °C were the best conditions for amylase production in black fonio grain while pH 6.0 and 20 °C were obtained for white fonio grains. The study concluded that grains of white and black fonio are rich sources of amylases and they could be of high value in various biotechnological applications.

Genomic DNA Extraction Methods for Optimal Quality DNA Recovery from Tomato Yellow Leaf Curl Virus (TYLCV)

Elizabeth G. Panerio — 2025-09-01

High quality DNA is essential for numerous downstream applications, especially PCR. Currently, various protocols for plant DNA extraction already exists, many of which are modifications of the CTAB method, aiming to enhance the extraction process and improve the quality and yield of DNA. This study primarily focuses in comparing three DNA extraction protocols commonly used to detect Tomato Yellow Leaf Curl Virus on infected tomato plants. Ten tomato leaf samples that were treated with liquid nitrogen were used in the experiment. Based on the results, the CTAB method has the highest DNA concentration mean of 1718.01ng/µL and it’s the only protocol that passed the purity ratio of A260/280 with an average value of 1.96 (range: 1.74-2.12) and a value of 1.45 on purity ratio of A260/230. Interestingly, despite its superior DNA quality, the CTAB method had the lowest number of samples that successfully amplified the A2 gene. This occurrence needs further investigation to identify what causes the contrasting results. In this case, the commercial kit and the NaOH produced better results in amplifying the viral A2 gene of the Tomato Yellow Leaf Curl Virus. Further optimization on the DNA extraction protocol is highly encouraged to produce high qualityDNA extract that will effectively amplify the viral gene and produce distinct PCR bands.

Effect of Different Substrates on Composition and Quality of Vermicompost

Pranali Ramteke — 2025-09-02

A field experiment was conducted during September 2024 to March 2025 at the Vermicompost Unit, Division of Soil Science, College of Agriculture, Pune, to evaluate how different organic substrates, in combination with microbial consortia, affect the composition and quality of vermicompost. We hypothesized that substrate type, when integrated with microbial consortia, would significantly influence nutrient enrichment, microbial activity, and heavy metal content in the final vermicompost product. The experiment followed a completely randomized block design with seven treatments—tree litter, button mushroom spent compost, wheat straw, soybean straw, coconut coir, sugarcane trash, and farmyard manure—each replicated three times. Defined microbial consortia were introduced during composting to enhance microbial-mediated decomposition. Vermicompost derived from tree litter showed superior quality, recording the highest organic carbon (23.96%), total nitrogen (1.93%), available phosphorus (0.68%), and potassium (1.25%). Micronutrient concentrations (Fe: 1575.07 mg kg⁻¹, Mn: 426.38 mg kg⁻¹, Zn: 299.30 mg kg⁻¹, Cu: 184.95 mg kg⁻¹) were also highest in the tree litter treatment. Additionally, heavy metal content (Cd: 0.28, Pb: 0.49, Cr: 0.36 mg kg⁻¹) was lowest in this treatment. Enhanced microbial enzyme activities—including dehydrogenase (34.90 μg TPF g⁻¹ 24 hr⁻¹), acid phosphatase (3.84 μg PNP g⁻¹ 2 hr⁻¹), and urease (118.42 μg NH₄⁺-N g⁻¹ day⁻¹)—further confirmed improved biological quality. Tree litter vermicompost also exhibited lower bulk density (0.73 g cm⁻³), finer particle size (<4 mm), and optimal color (very dark greyish brown, 10YR3/2). These results suggest that tree litter, when combined with microbial consortia, serves as an optimal substrate for high-quality vermicompost production, with enhanced nutrient content, reduced contaminants, and improved biological activity—offering practical benefits for sustainable agriculture.

Influence of Organic Substrates and Microbial Consortia on the Physicochemical Properties and Nutrient Profile of Vermiwash

Pranali Ramteke — 2025-09-04

A field experiment was conducted during 2024 at Vermicompost Unit, Division of Soil Science, College of Agriculture, Pune to study the effect of different substrates along with microbial consortia on composition and quality of vermiwash. The experiment was laid out in completely randomized block design having seven treatments with three replications. The treatments comprised T₁-tree litter, T₂-button mushroom spent compost, T₃-wheat straw, T₄-soyabean straw, T₅-coconut coir, T₆-sugarcane trash and T₇-farm yard manure for preparation of vermiwash. The findings of the present investigation revealed that all treatments showed a gradual increased in pH from the 16^th to 48^th day of harvesting. The significantly higher N, P and K contents were noted in tree litter vermiwash (0.8, 0.74, and 0.5 % respectively). On the 16^th day the treatment tree litter recorded higher dehydrogenase activities (18.64 µg TPF ml⁻¹ 24 hr⁻¹). Urease activity was higher in tree litter on both 16^th (10.50 µg NH₄⁺-N ml⁻¹ day⁻¹) and 32^nddays (24.50 µg NH₄⁺-N ml⁻¹ day⁻¹). The vermiwash produced from tree litter substrate showed higher macronutrient.

An Assessment of the Effect of Azotobacter on the Mineral and Vitamin Content of Amaranthus viridis

Ujah II — 2025-09-09

Amaranthus viridis is a vegetable widely grown and consumed in Nigeria. There is the need for a sustainable approach that will enhance its cultivation and growth. This study was conducted in response to the growing concern about the insufficient supply of essential minerals and vitamins in vegetables, which contributes to nutrient deficiencies in human diets. The research aimed to assess the effect of Azotobacter treatment on the mineral and vitamin content of Amaranthus viridis. The study involved treating Amaranthus viridis plants with varying concentrations of Azotobacter (100g, 200g, 300g, 400g, viable and very viable), and comparing them with the control. The results revealed that at 400g inoculation, nitrogen increased by 98%, magnesium by 320%, calcium by 686%, and copper by a massive 8575% compared to the control.Some treatments however reduced nutrients, phosphorous at 200g dropped by 82% compared to the untreated one control. Vitamin B1 showed its greatest improvement at 400g, with an increase of 55.6%, while the Viable treatment caused a slight reduction. Vitamin B2 was boosted most strongly under the Viable treatment, recording a 50% increase, though 400g and 300g also gave substantial gains. For Vitamin C, the highest effect was observed at 400g with a 57.9% increase, followed by Very Viable with 34.2%. Vitamin E recorded the most remarkable response, with a 300% increase at 400g and 200% under Very Viable. The findings suggest that higher concentrations of Azotobacter positively influence the mineral and vitamin content in Amaranthus viridis, enhancing its nutritional quality. This study shows that Azotobacter can be used as a biofertilizer to improve the mineral profile of leafy vegetables, contributing to better human health through improved dietary intake and in sustainable agriculture.

Residual Effect of Different Farming Practices on Soil Biochemical Properties under Wheat Cultivation

Rutuja Chaukhe — 2025-09-09

Field experiment was conducted to study residual effect of farming practices soil biochemical properties under wheat cultivation at Agronomy Farm, College of Agriculture, Pune during kharif, 2024. Research was conducted as a sequence cropping of soybean-wheat which was started during 2022-23. Randomized Block Design (RBD) with four replications with five treatments namely conventional practices (T₁), GRDF (T₂), organic farming (T₃), zero budget natural farming (T₄) and climate resilient farming (T₅). The results revealed that, the chemical properties like pH and EC were not significantly influenced under different farming practices after harvest of wheat. However, the organic farming registered significantly higher organic carbon (0.75 %) which is closely followed by climate resilient farming (0.74) and GRDF (0.71 %) which were par with each other. Calcium carbonate content 9.33 % was a significantly lower under organic farmers than other farming practices. observed in organic farming (9.33%) and GRDF (9.37 %). The highest available nitrogen (191.18 kg ha^-1), available phosphorus (28.36 kg ha^-1), available potassium (484.95 kg ha^-1) and available sulphur (18.75 kg ha^-1) recorded in climate resilient farming. While, soil available micronutrient was higher under organic farming viz., Fe (6.53 mg kg^-1), Mn (3.49 mg kg^-1), Zn (2.98 mg kg^-1) and Cu (3.71 mg kg^-1) respectively. The organic farming practice found beneficial in respect to microbial population such as Bacteria, fungi and actinomycetes with value (207.21 x 10⁶ cfu g^-1soil, 23.91 x 10⁵ cfu g^-1soil and 48.20 x 10⁴ cfu g^-1soil) and enzymatic activities, urease (33.16 µg NH₄ ⁺-N g^-1 soil 24hr^-1), dehydrogenase (15.26 µg TPF g^-1 soil 24 hr^-1), acid phosphatase (13.24 µg PNP g^-12 hr^-1) and alkaline phosphatase (14.76 µg PNP g^-12 hr^-1). Organic and climate-resilient farming sustain soil health and fertility, ensuring productive and eco-friendly wheat cultivation.

GPS-Based Soil Fertility Mapping of Agriculture College Farm, Pune, Maharashtra, India

Tejal Pawar — 2025-09-13

Present soil survey was carried out during 2024 in College of Agriculture, Pune (Maharashtra) India by using Global Positioning System (GPS). Fertility maps were prepared with 160 geo-referenced soil samples collected by randomized method. Out of 160 soil samples, 127 samples from Agronomy Farm and 33 samples were collected from Modi Baugh and Pathology Farm by using standard soil sampling procedure and were analysed for chemical properties at College of Agriculture, Pune with an objective to assess the soil nutrient status and delineate soil fertility maps of Agriculture College farm, Pune. The pH and EC of soils of Agronomy Farm was found in between 7.63 to 8.96 (mean 8.11) and 0.20 to 0.78 dS m^-1 with mean 0.41 dS m^-1 respectively. The organic carbon and calcium carbonate content of the Agronomy Farm was ranged from 0.10 to 0.73 per cent with mean 0.39 per cent and 10.25 to 27.5 per cent with mean 18.53 per cent respectively. The available nitrogen, phosphorous and potassium of Agronomy Farm was found to be between 75.27 to 326.11 kg ha^-1(mean 195.31 kg ha^-1), 6.57 to 60.08 kg ha^-1(mean 31.82 kg ha^-1) and 114.24 to 1005.76 kg ha^-1 (mean 422.24 kg ha^-1) respectively. The available sulphur ranged between 16.21 to 26.46 mg kg^-1 (mean 21.44 mg kg^-1).

The soils of Modi Baugh and Pathology Farm showed pH and EC from 7.52 to 8.45 with mean 7.85, and 0.21 to 0.64 dS m^-1 with mean 0.45 dS m^-1 respectively. The organic carbon content and calcium carbonate ranged from 0.17 to 0.62 per cent with mean 0.32 and 8.5 to 25.5 per cent with mean 14.1 per cent respectively. The available nitrogen, phosphorous and potassium of Modi Baugh and Pathology Farm ranged from 125.44 to 301.06 kg ha^-1 (mean 222.76 kg ha^-1), 13.14 to 31.6 (mean 20.17 kg ha^-1) and 166.72 to 919.52 kg ha^-1 (mean 400.10 kg ha^-1)respectively. The available sulphur in soils ranged between 18.50 to 25.39 mg kg^-1 (mean 22.31 mg kg^-1).

Based on the GPS locations of samples and soil test results, soil fertility maps of Agriculture College Farm, Pune were prepared by using Arc-GIS 10.8.2 for soil pH, EC, OC, CaCO₃, available N, P, K and S.

Comparative Assessment of Farming Practices on Wheat Yield, Quality Parameters and Nutrient Uptake

Rutuja Chaukhe — 2025-09-18

A field experiment was conducted at the Agronomy Farm, College of Agriculture, Pune during kharif, 2024 to study residual effect of farming practices on yield, quality and nutrient uptake under wheat cultivation. The experiment was initiated in 2022- 23 as a soybean-wheat sequence cropping system. A randomized block design (RBD) four replications and five treatments was adopted, comprising coventional practices (T₁), General recommended dose of fertilizers (T₂), organic farming (T₃), zero budget natural farming (T₄) and climate resilient farming (T₅). The results revealed that, climate resilient farming (T₅) recorded significantly higher grain yield (43.28 q ha^-1) and straw yield (59.72 q ha^-1). Quality parameters such as protein content (12.60 %) and hectolitre weight (81.91 kg hl^-1) were also significantly superior under climate resilient farming. Nutrient uptake was enhanced under T₅, with significantly higher total N (128.79 kg ha^-1), P (56.42 kg ha^-1), and K (115.46 kg ha^-1) uptake compared to other treatments. Morever, uptake of micronutrients by wheat was also highest under T₅, with Fe (1161.47 g ha^-1), Mn (431.5 g ha^-1), Zn (282.33 g ha^-1) and Cu (60.36 g ha^-1) r closely followed by General recommended dose of fertilizer.

Dietary Exposure and Health Risk Assessment of Aflatoxin B1 in Stored Groundnut (Arachis hypogaea) and Maize (Zea mays) Samples of Katsina State, Nigeria

Abubakar R — 2025-09-19

Aflatoxin B₁ contamination in food is a significant global health challenge, particularly affecting children, leading to delayed development, stunted growth, liver damage, and liver cancer. The study was carried out in 2024 in Katsina State, Nigeria. A study aimed to determine the levels of aflatoxin B₁ and attendant health risks to the population from consumption of stored groundnut and maize samples in the three senatorial zones of Katsina State (Katsina, Daura, Funtua). The quantification of Aflatoxins in the samples was conducted using Enzyme-Linked Immunosorbent Assay (ELISA). The growth of fungal species was determined using the spread-plate method after dilution. Risk characterization was conducted based on the margin of exposure (MOE) and the liver cancer approach. The results revealed that 90% and 95% of Groundnut and Maize samples collected from all three geopolitical zones of Katsina State were contaminated with either Aspergillus flavus or Aspergillus parasiticus, respectively. From the results, all the samples contain aflatoxin B₁ levels above the permissible limit of Nigeria (20µg/kg), the World Health Organization (WHO) and the European Union (EU) (4µg/kg). There was no significant difference in the mycological analysis of the samples in all three zones of the State. The calculated MOE values were lower than 10,000, pointing to a public health risk that may arise due to AFB₁ exposure through the consumption of the study samples.

Comparative Proximate Analysis of Piper guineense Leaf and Xylopia aethiopica Seed

Ubah, V.C.S. — 2025-09-22

Piper guineense leaf and Xylopia aethiopica seeds are two plant materials commonly used in traditional medicine and as spices in cooking, particularly in South-Eastern Nigeria and other parts of Africa, due to their high nutritional properties. Nursing mothers in these regions often consume soups prepared with these plants, believing they offer nutritional and health benefits. This study was aimed at evaluating the proximate composition of crude extract from Piper guineense leaf and Xylopia aethiopica seeds. In this study, the proximate analyses were identified, quantified and evaluated using standard protocols. The sample was heated at 100-150 °C for a specific time until it reached a constant weight. The crude fibre content was determined by defatting about 2g of material with petroleum ether (if the fat content exceeded 10%). Crude Fibre, Crude Fat and Crude Proteins were determined. The result on determination of the proximate composition of the samples showed that, Xylopia aethiopica contains crude protein (8.32±0.46), fat (7.53±0.35), carbohydrate (59.23±4.25), ash content (12.89±0.71), moisture (13.44±0.27), fibre (1.17±0.37), Piper guineenses contains crude protein (9.26±0.99), fat (4.02±0.43), carbohydrate (57.13±6.04), ash content (10.54±0.38), moisture (18.53±0.19), fibre (3.22±0.44). These plants can contribute important nutrients to diets. These findings indicate that the spices are notable wellsprings of bioactive components and justify their plethoric applications in Nigeria. Therefore, they could serve as lead compounds in the search for natural ingredients for drugs and nutraceutical formulations.

Investigating the Relationship Between Soil Enzyme Activities and Soil Nutrients

A. M. Memane — 2025-09-22

Soil enzymes mediate the transformation of organic matter into nutrient-rich compounds and play a central role in energy transfer. Their activity influences the recovery of organic matter, soil structure, microbial biomass and overall microbial activity. The present study was conducted during January, 2023 to April, 2025 at Post Graduate laboratory, College of Agriculture, Pune with an objective to find out relationship between soil enzyme activities with soil nutrients of sugarcane ratoon. The study focused on dominant sugarcane growing districts in Maharashtra state, under different sugar recovery zones viz., high, medium and low. A total of 180 farmers were selected from these districts. Among these 90 farmers followed long term in-situ sugarcane trash management system while the other 90 adopted long term in-situ sugarcane trash burning system, which they had been implementing for the past ten years. After harvest of sugarcane ratoon, the soil samples collected from selected sites at the depth of 0-30 cm where the active root zone of sugarcane exist and related data was collected during the year 2022-23 through a structured survey, ensuring a thorough understanding of the farming practices and their impact on soil biological properties.

Furthermore, a correlation analysis was performed to examine the relationships between soil enzyme activities and soil nutrients. Enzyme activities viz., dehydrogenase, urease, acid phosphatase, alkaline phosphatase and β-glucosidase were positively correlated with organic matter in both sugarcane trash management and burning practices. Meanwhile, urease activities showed a positive correlation with nitrogen. Acid and alkaline phosphatase showed a positive correlation with phosphorus.

Nutritional, Phytochemical, and Antioxidant Profiling of Dysphania ambrosioides Leaves from Different Locations of Kangra Valley, Himachal Pradesh,India

Shivani Sharma — 2025-09-23

Medicinal plants have gained considerable attention in recent years due to their therapeutic potential and nutritional benefits. In this study, the proximate composition, mineral content, phytochemical profile, and antioxidant activity of Dysphania ambrosioides leaves were analyzed from twelve locations across the Kangra Valley, Himachal Pradesh, India. Considerable variation was observed in nutritional traits (moisture 26-37%, protein 23-31%, fat 3-7%, fibre 9-11%, carbohydrates 5-14%) and mineral concentrations (sodium 228-315 mg/100 g, calcium 913-1025 mg/100 g, potassium 983-1087 mg/100 g). Bioactive compounds also differed notably, including phenolics, tannins, chlorophyll, carotenoids, and vitamin C, alongside antioxidant activity. Collections such as Dehra, Harchkiyan, Jaswan, Palampur, Daroh, and Thural demonstrated superior nutritional and phytochemical attributes. This work represents the first comparative profiling of D. ambrosioides from different locations of Kangra Valley and underscores its potential as a nutritionally dense, antioxidant-rich species with promising applications in functional foods, nutraceuticals, and herbal formulations.

Neem-Mediated Silver Nanoparticles: Characterization and Biological Applications against Mosquito Larvae and Pathogenic Bacteria

Titilayo Oluwaseun AGUNBIADE — 2025-09-28

Introduction: Nanotechnology offers sustainable solutions for healthcare and environmental management. Silver nanoparticles (AgNPs) were green-synthesized using Azadirachta indica (Neem) leaf extract, a plant renowned for its broad pharmacological activities.

Aim: To biosynthesize neem-mediated AgNPs, characterize them physicochemically, and evaluate their larvicidal and antibacterial potency.

Materials and Methods: Aqueous neem leaf extract was mixed with AgNO₃; formation was tracked by colour change and UV–Vis spectroscopy (458–459 nm). FTIR identified functional groups, SEM measured morphology/size, and standard bioassays determined larvicidal activity against Culex quinquefasciatus larvae and antibacterial activity against selected pathogens.

Results: UV–Vis confirmed AgNP formation; FTIR showed hydroxyl, carbonyl and amine groups acting as reducing/stabilizing agents. SEM revealed 20–50 nm predominantly spherical particles. Probit analysis gave 24-h LC₅₀ values of 0.0279 mg L⁻¹ (AgNPs) versus 0.1985 mg L⁻¹ (neem extract). Agar-well diffusion assays showed larger inhibition zones for AgNPs than for extract, strongest against Staphylococcus aureus.

Conclusion: Neem-mediated AgNPs were successfully synthesized and characterized, showing strong larvicidal and antibacterial activities. These results highlight their potential as eco-friendly agents for vector control and antimicrobial use. Further in vivo and environmental safety studies are needed to confirm their applicability.

Evaluation of the Renal Function Response to Aqueous Leaf Extracts of Selected Edible Plants in Male Wistar Rats

Temioluwa E. OSOLAJA — 2025-09-28

This study evaluated the renal function response to aqueous leaf extracts of five commonly consumed edible plants—Solanum aethiopicum (SA), Amaranthus hybridus (AH), Pterocarpus mildbraedii (PM), Ocimum gratissimum (OG), and Telfairia occidentalis (TO)—in male Wistar rats over a 60-day exposure period. Rats were administered low, medium, and high doses of each extract orally, and serum biochemical markers, including blood urea nitrogen (BUN), creatinine, bicarbonate, sodium, potassium, and chloride, were assessed. Relative to the control, BUN levels decreased slightly with SA (23.40 ± 0.25 mg/dL at high dose) but increased with AH (25.30 ± 0.30 mg/dL), OG (26.10 ± 0.50 mg/dL), and TO (26.40 ± 0.28 mg/dL). Creatinine showed a modest decline with SA (0.32 ± 0.03 mg/dL) but increased with AH (0.45 ± 0.04 mg/dL), OG (0.48 ± 0.05 mg/dL), and TO (0.47 ± 0.04 mg/dL). Potassium levels rose slightly with SA (25.90 ± 0.40 mmol/L) and declined with AH (24.60 ± 0.35 mmol/L), PM (24.70 ± 0.32 mmol/L), OG (22.50 ± 0.35 mmol/L), and TO (24.00 ± 0.32 mmol/L). Chloride was minimally affected by SA, AH, and PM but decreased markedly with OG (65.00 ± 1.00 mmol/L) and moderately with TO (68.20 ± 1.00 mmol/L). Two-way ANOVA indicated that vegetable type significantly influenced potassium (F = 11.411, p < 0.001, η² = 0.695) and chloride levels (F = 3.074, p = 0.040, η² = 0.381), while dose interactions were significant for potassium (F = 2.721, p = 0.023, η² = 0.620). Overall, these results demonstrate that the leaf extracts exert extract-specific and dose-dependent effects on renal biochemical markers, highlighting both potential nephroprotective and nephroactive properties. The study underscores the importance of cautious and informed use of edible and medicinal plant extracts.