Research Perspective on Biological Science Vol. 11

Ion-Exchange Chromatography-Based Purification and Characterisation of Isocitrate Lyase Isoforms from Corn Scutella: Insights into Enzyme Function

Saba Hadi — 2026-04-15

Background: Isocitrate lyase is widespread in bacteria, mushrooms and higher plants. In metazoans, the enzyme is present in some nematodes during the adult stage, post- embryonic larvae and embryos and probably in some arthropods. The presence of ICL and the glyoxylate cycle in vertebrates remains debated.

Aim: The purpose of this study is to devise a method for separating isocitrate lyase isoenzymes from corn scutella by using ion-exchange chromatography.

Methods: The isocitrate lyase enzyme was purified from corn plants using ion exchange chromatography and sulfate precipitation techniques. Enzyme activity was assessed using spectrophotometric methods, and the molecular weight was determined through gel chromatography. Studies were conducted to investigate the influence of pH, glycine, and glycolate on enzyme activity. The specific identification of the enzyme was determined using a modified Schiff reagent. SDS- PAGE electrophoresis was performed with a 12.5% polyacrylamide gel concentration. Standard marker proteins (Sigma) were used to plot the calibration curve.

Results: Two isoforms of isocitrate lyase, ICL1 and ICL2, were purified, exhibiting molecular masses of 164 kDa and 208 kDa, respectively. ICL1 demonstrated optimum activity at pH 7.5, while ICL2 exhibited optimum activity at pH 6. In this study, specific concentrations of glycine and glycolate were found to enhance the enzymatic activities of both isoforms. A highly statistically significant result was obtained using the P – value of less than.05. Post-hoc Tukey's test showed that the specific activity of ICL2 was significantly higher than that of ICL1, and the Km value of ICL2 was significantly greater than that of ICL1 (P < .05). These results further support the hypothesis that the two isoforms of ICL have distinct functional roles in metabolic processes.

Conclusion: This research provides significant insights into the characteristics of the isocitrate lyase enzyme in corn plants. The data indicate the presence of distinct enzyme forms with specific interactions in varying environmental conditions, which may be applicable in agricultural practices to increase crop yield and improve the metabolic turnover of organic acids.

Enhancing Immune Response in Local Chickens Using Fermented Snail Meat Hydrolysate Under Low-Protein Feeding Conditions

Ujang Suryadi — 2026-04-15

Background: Protein is a critical component in poultry diets, essential for growth, muscle development, and overall production efficiency. However, high protein consumption causes a low protein efficiency ratio and causes feed prices to be high.

Objective: This study aimed to investigate how fermented snail meat hydrolysate modulates both the production performance and immune systems of local chickens.

Materials and Methods: The research used snail meat hydrolysate resulting from submerged fermentation with starter culture from rice washing water. Using hydrolysates as a fishmeal substitute to reduce the protein content of male local chicken diets. Hydrolysate is mixed into the formulated ration: P0 = 12% fish meal in feed without hydrolysate added, P1 = 8% fish meal in feed plus 5 ml hydrolysate/kg feed; P2 = 4% fish meal in feed plus 10 ml hydrolysate/kg feed; and P3 = Feed without fish meal plus 15 ml hydrolysate/kg feed. The study used completely randomised study parameters, namely the production performance of 200 chickens and the immune system (including lymphoid organs and haematological status) of 60 chickens.

Results: Incorporating fermented snail meat hydrolysate into chicken feed in place of fish meal showed variable effects on production performance. Feed consumption remained statistically unchanged (p > 0.05) across treatments, indicating that replacing fish meal with snail meat hydrolysate did not affect feed intake. Substituting fish meal with 10 ml/kg of snail meat hydrolysate also had no significant effect (p > 0.05) on final body weight gain. These findings suggest that snail meat hydrolysate (10 ml/kg) can replace the reduction in the use of fish meal from 12% to 4% in ration formulation without reducing production performance and has no negative effect on the immune system.

Conclusion: Snail meat hydrolysate has the potential to be used as an immune antibiotic to replace the use of fishmeal in the development of low-protein dietary formulations without affecting production performance or negatively affecting the chicken's immune system.

Behavioural Responses of Bagworm Larvae to Biotic and Abiotic Environmental Factors: A Comprehensive Review

Janhiphula Kanhar — 2026-04-15

Bagworm larvae (Lepidoptera: Psychidae) display diverse behavioural adaptations that enable survival under varying biotic and abiotic environmental conditions. This review summarises key behavioural responses, including host plant selection, dispersal, phototactic movement, silk-assisted locomotion, and protective case construction. Environmental factors such as light, climate, and habitat structure influence larval movement and development, while interactions with predators and natural enemies shape defensive and foraging behaviours. The portable bag, supported by strong silk and unique fibroin genes, serves both as protection and as an adaptive tool for survival. Studies also highlight the role of ecological interactions and population dynamics in pest outbreaks and crop losses, particularly in oil palm ecosystems. Recent genomic and behavioural research provides deeper insights into mechanisms underlying larval adaptability. Overall, understanding these behavioural responses is essential for improving ecological knowledge and developing sustainable pest management strategies.

Molecular Assessment of Clonal Fidelity and Efficient Somatic Embryogenesis in Kaempferia marginata (Carey ex Roscoe): An Endangered Medicinal Species

Chongtham Henary Singh — 2026-04-15

Kaempferia marginata Carey ex Roscoe (Zingiberaceae) is a small perennial herb that belongs to the Zingiberaceae family. The present study aims to standardise a highly efficient and reproducible in vitro micropropagation protocol via somatic embryogenesis for Kaempferia marginata, an endangered and medicinally significant herb, and to concurrently validate the genetic fidelity of the regenerated plantlets. Because traditional propagation via rhizomes is extremely slow and susceptible to soil-borne pathogens, somatic embryogenesis was selected to provide a high-frequency, scalable alternative. Sprouting rhizome buds were utilised as explants and cultured on Murashige and Skoog (MS) basal medium. To evaluate callus induction and subsequent shoot multiplication, the media were supplemented with various concentrations of plant growth regulators (PGRs), including α-naphthalene acetic acid (NAA), 2,4-Dichlorophenoxyacetic acid (2,4-D), indole acetic acid (IAA), 6-benzylaminopurine (BAP), and Kinetin. Furthermore, to assess the genetic uniformity of the in vitro regenerants against the mother plant, genomic DNA was extracted, and molecular profiling was conducted using Random Amplified Polymorphic DNA (RAPD) and Inter Simple Sequence Repeat (ISSR) markers. The highest frequency of embryogenic callus induction (87.00 ± 0.10%) was achieved on MS medium fortified with 2.0 mg L⁻¹ NAA. Following the developmental stages of somatic embryos (globular to torpedo), transfer to regeneration media revealed that a synergistic combination of 4.0 mg L⁻¹ BAP and 1.0 mg L⁻¹ NAA yielded the maximum number of shoots (16.35 ± 0.22 shoots per explant). Molecular analysis utilising 15 RAPD primers and 11 ISSR primers generated 84 and 76 scorable bands, respectively. The banding patterns across all 15 randomly selected regenerants showed no detectable polymorphism using RAPD and ISSR markers, indicating high genetic stability. The absence of somaclonal variation confirms that this optimised somatic embryogenesis protocol provides a reliable and scalable method for the mass propagation of true-to-type elite clones. This offers a highly effective biotechnological tool for both the sustainable commercial supply of pharmaceutical raw materials and the ex situ conservation of K. marginata germplasm.

The Independent Contribution of Body Mass Index and Sex to Blood Pressure Variance in a Large Cohort of Young Adults: A Cross-Sectional Study

Taranjot Kour — 2026-04-15

Background: Hypertension and obesity are increasingly reported in young adults, particularly in rural India. Early assessment of cardiometabolic markers is essential for prevention.

Aim: To evaluate anthropometric measures, blood pressure, and muscle strength in rural young adults and examine sex-based differences and BMI–BP associations.

Methods: A cross-sectional study was conducted among rural young adults. Height, weight, BMI, BP indices (SBP, DBP, MAP, pulse pressure), and hand-grip strength were recorded. BP categories were assigned using standard guidelines. Correlation and regression analyses assessed relationships between BMI, sex, and BP.

Results: Males had higher BP and grip strength despite similar BMI. Nearly half were pre-hypertensive and ~30% hypertensive, with greater prevalence in males. BMI correlated positively with SBP, DBP, and MAP. Regression showed BMI and male sex as independent predictors of BP.

Conclusion: Young adults in rural communities show early signs of elevated BP, strongly associated with BMI and sex. Early screening and lifestyle-focused preventive strategies are essential to reduce future cardiovascular risk.

Assessment of Haematological Changes in Clarias batrachus Following Exposure to Sago Factory Effluent

Ramesh F — 2026-04-15

Haematological indices are important parameters to evaluate the general physiological status of fishes and may be considered as stress indicators for estimation of the response reactions of the fish to various environmental conditions. This chapter explores the impact of sago industry effluent on the blood characteristics of the freshwater fish Clarias batrachus, with emphasis on changes that occur during prolonged exposure. The continuous release of agro-industrial waste into aquatic environments has become a growing concern, particularly due to its potential effects on fish health. In this context, haematological parameters provide a practical and sensitive means of assessing physiological stress in aquatic organisms. In the present study, fish were maintained in different concentrations of sago effluent (0%, 25%, 50%, and 75%) for a period of 28 days under sublethal conditions. At the end of the exposure period, clear alterations in blood parameters were recorded. There was a gradual reduction in red blood cell count, haematocrit, mean corpuscular volume, and mean corpuscular haemoglobin as the effluent concentration increased. These changes point towards a reduced capacity for oxygen transport and possible interference with normal blood cell formation. On the other hand, an increase in white blood cell count was observed, which may reflect a defensive response to stress. Overall, the study indicates that sustained exposure to sago effluent can disrupt normal physiological functions in fish. The observed variations in blood indices highlight their usefulness as early indicators of environmental stress. The findings also stress the importance of effective treatment and management of industrial effluents to minimise their impact on aquatic ecosystems.

In vivo and in vitro Assessment of Antioxidant Activity of Culcasia falcifolia Leaf Extract

A. Gracelyn Portia — 2026-04-15

Introduction: Epilepsy is one of the common neurological disorders that is characterised by repetitive seizures. Oxidative stress, free radical production and membrane lipid peroxidation occur as a result of epilepsy, which causes damage to the brain tissue. Seizure generation can be associated with the homeostatic imbalance between antioxidants and oxidants. Oxidative stress is the most prominent mechanism in the development and progression of epilepsy. It has been known that during an epileptic attack, oxidative stress occurs, free radicals are produced, and membrane lipid peroxidation happens, all of which cause tissue damage.

Aim: The study aims to evaluate the in vitro and in vivo antioxidant potential of the ethanol leaf extract of Culcasia falcifolia against pentylenetetrazole-induced oxidative stress.

Method: The extract was prepared by maceration using ethanol. Four groups of six mice were used for the in vivo studies. Group I (control group) received 0.1% CMC. Group II (positive control) received diazepam on the experimental day (5mg/kg body wt.). Group III and IV received 200 and 400 mg/kg body wt. of ethanol extract of C. falcifolia orally for fourteen days. On the fifteenth, all mice were sacrificed to remove the brains after the injection of PTZ (60 mg/kg i.p). MDA, SOD, CAT, GPX and GR levels were estimated from the brain tissue as part of the in vivo assessment. DPPH radical scavenging assay and ferrous ion chelating assay.

Result: The study suggests that the extract exhibits in vivo antioxidant property by increasing the levels of SOD, CAT, GPX and GR and decreasing the MDA level of the brain tissue significantly (p<0.05*). The in vitro studies also showed that the extract effectively scavenged free radicals and chelated ferrous ions, indicating strong antioxidant potential.

Conclusion: Both in vivo and In-Vitro studies demonstrate that the ethanol extract of Culcasia falcifolia possesses antioxidant activity against Pentylentetrazole-induced seizure in mice. These findings show that the extract has the ability to attenuate the oxidative stress caused by PTZ-induced seizure. The extract likely contains active phytochemicals, which may be responsible for in vivo and in vitro antioxidant activity against PTZ-induced seizure in mice. Further studies are needed to isolate the active compound to evaluate its therapeutic efficacy in epilepsy.

Cattle Identification and Management Using Nose Muzzle Print Biometrics: A Review

J. N. Sreedhara — 2026-04-15

Individual feedlot beef cattle identification represents a critical component in cattle traceability in the supply food chain. It also provides insights into tracking disease trajectories, ascertaining ownership, and managing cattle production and distribution. Historically, cattle identification relied on conventional methods such as ear cutting, hot iron branding, tattooing, ear notching, and manual sketching of coat colour patterns for registration purposes. However, many of these traditional techniques do not align with animal welfare standards, often causing stress to animals. In addition, they are prone to inaccuracy, poor durability, and potential loss or damage. This review provides a comprehensive examination of cattle identification through nose (muzzle) prints, a highly promising biometric application. It delves into the unique dermatoglyphic patterns of bovine muzzles, comparing their inherent uniqueness. The report traces the evolution of identification methodologies from traditional, often invasive, physical impression techniques to advanced digital approaches leveraging sophisticated image processing, machine learning, and deep learning algorithms. Deep learning image models taken with a mirrorless digital camera and processed to form the dataset will be of great significance in muzzle identification with 98.7% accuracy. It highlights the significant advantages of muzzle print biometrics, including their non-invasive nature, contribution to animal welfare, and high accuracy in diverse applications such as disease control, traceability, and theft prevention. Furthermore, the review explores the challenges in real-world implementation, particularly concerning environmental factors and animal movement, and projects future advancements, emphasising the synergistic integration of muzzle print technology within broader smart farming ecosystems encompassing the Internet of Things (IoT), predictive analytics, and blockchain. The major concern was on the proper identification of cattle for registration and of cattle on an official test so that the possibility of swapping, false insurance claims and ownership disputes can be guarded against. This technique will certainly help in livestock census to avoid duplication and also be helpful in geographical tagging of specific breeds, as no two breeds have the same nose muzzle pattern, and it can be compared to human fingerprints.

Impact of Neem (Azadirachta indica) and Papaya (Carica papaya) Leaves on Staphylococcus aureus Virulence Factors

Aparna Shil — 2026-04-15

Staphylococcus aureus is a prominent causative agent across a spectrum of conditions, from superficial skin lesions to invasive diseases and remains difficult to manage because of antimicrobial resistance and biofilm-associated tolerance. This study evaluated aqueous and ethanolic leaf extracts of Neem (Azadirachta indica) and Papaya (Carica papaya) for anti-virulence activity against S. aureus ATCC 25923. Bacteria were exposed to extracts (125 μg/mL) in tryptic soy broth; growth (metabolic activity) was quantified spectrophotometrically. Biofilm formation on glass plates (60 mm) was assessed by a crystal violet–acetic acid assay, and haemolysis was evaluated on blood agar containing 7% sheep blood. Antibacterial activity of aqueous and ethanolic extracts (125–250 μg/mL) was examined by disc diffusion on Mueller–Hinton agar. Data were analysed by one-way ANOVA with Dunnett’s multiple comparisons test (p < 0.05). Neem reduced growth relative to the untreated control, whereas Papaya alone did not reduce growth at the tested concentration. Both extracts reduced biofilm formation, with ethanolic extracts showing stronger inhibitory effects in disc diffusion (15.6 mm for Neem and 14.0 mm for Papaya at 250 μg/mL; antibiotic control ~17–18 mm). None of the extracts altered haemolytic activity under the conditions tested. These findings suggest Neem and Papaya leaf extracts can attenuate S. aureus biofilm formation and exhibit modest antibacterial activity, supporting further fractionation and mechanistic studies.

Larvicidal and Adult Emergence Inhibition Effects of Myrtus communis and Eucalyptus globulus Leaf Extracts against Culex molestus

Sarah Kadhim Al-Rahimy — 2026-04-15

Background: Mosquito-borne diseases have become increasingly severe due to rapid urbanisation and population growth. Although chemical insecticides have been widely used for mosquito control, their extensive application has led to problems such as insecticide resistance and environmental pollution. Therefore, there is growing interest in plant-based alternatives as eco-friendly and effective mosquito control agents.

Objective: The current research aimed to determine the effects of aqueous and alcoholic extracts of Eucalyptus globulus and Myrtus communis on adult mosquito emergence inhibition and the mortality rate of fourth-instar larvae.

Material and Methods: This study was conducted in Karbala Governorate, Iraq, in March 2021, to control the Culex molestus (Diptera: Culicidae) mosquito, calculate the mortality rate of the fourth larval stage, and inhibit the emergence of the adult insect. Concentrations of the aqueous and hexane extracts were (0.2, 0.4, 0.6, 0.8, and 1.0) mg/ml each, while the concentrations of the cold aqueous extract of M. communis were (0.5, 1, 1.5, 2, and 2.5) mg/ml. The concentrations were (0.5, 1, 2, 4, 8) (mg/ml) for the boiled and cold water extracts, while the concentrations were (0.1, 0.2, 0.3, 0.4 and 0.5) mg/ml for the hexane extract of E. globulus plants. In addition, FTIR spectroscopy was used to identify the functional groups present in the plant extracts.

Results: The results of this study showed that the effect of the extracts varied depending on the concentrations, extraction method, and solvent used. This confirmed an increase in the mortality rate of the fourth larval stage and inhibition of adult emergence, as the extracts of the plants had the greatest effect in eliminating the fourth larval stage and inhibiting the emergence of adults by 100%, as these extracts outperformed the effect of eucalyptus extracts in the percentage of fourth larval stage mortality and inhibition of adult emergence when treated with cold and boiled water extracts and hexane, and eucalyptus extracts had an effect on the mortality of fourth instar larvae and inhibition of adult emergence compared to the control group treatments.

Conclusion: The results of the study showed that the extracts of the Myrtle and Eucalyptus plants are of great importance in controlling mosquitoes, which are medical insects that transmit many diseases. These extracts are natural, environmentally safe, harmless to non-target organisms, and quickly decompose. FTIR analysis confirmed the presence of bioactive functional groups responsible for their activity. The results provide evidence supporting the potential use of Eucalyptus globulus and Myrtle communis as natural pest control agents. These studies contribute to the broader scientific community by exploring environmentally friendly tools for mosquito control.

Morphological Characterisation, Genetic Validation, and Potential Development of 'Entog Kebumen': A Distinctive Superior Dwarf Coconut Variety from Indonesia

Ismail Maskromo — 2026-04-15

The demand for superior coconut varieties with early bearing and high productivity is still increasing. The Entog Kebumen dwarf coconut was a newly released variety; therefore, its stability needs to be analysed. This study presents the morphological characterisations and molecular analysis that underlie the official release of the ‘Entog Kebumen’ dwarf coconut as a superior variety from Indonesia. Observations conducted from 2016 to 2023 in Kebumen Regency, Central Java, evaluated the phenotypic stability and genetic distinctiveness of the population. The results confirmed that ‘Entog Kebumen’ was a unique dwarf coconut variety, characterised by its large fruit size and high fruit number, different from other dwarf coconuts. Other specific phenotypic markers were stiff leaves, round fruit shape, and light green exocarp. SSR analysis validated the genetic differentiation of the Entog Kebumen dwarf coconut from other previously released dwarf coconut cultivars. With 1,340 identified mother trees and a production capacity of 140,000 seeds per year (enough for 560 ha), this variety can be used for large-scale plantation development, especially for early-bearing coconuts, high-yielding, easy to harvest, and has a superior nutritional profile.