PhD Thesis
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Item Characterization and management of bacterial wilt of solanaceous vegetable in relation to physico-chemical and biological properties of soil(Department of Plant Pathology, College of Agriculture, Vellanikkara, 2025-11-15) Ajayasree, T S; Sible, George VargheseSolanaceous vegetables viz., tomato, chilli, and brinjal are widely cultivated in different agro-ecological units (AEUs) in Kerala. However, their cultivation is hampered by bacterial wilt disease. Being a soil-borne pathogen, the growth and survival of this bacterium may be greatly influenced by physico-chemical and biological properties of soil. Hence, the present investigation was carried out with the objectives of isolation and characterization of the bacteria responsible for bacterial wilt of solanaceous vegetables from four selected AEUs of Kerala; which were not studied earlier, to study the soil physico-chemical and biological properties influencing bacterial pathogen and to develop an integrated management strategy for bacterial wilt disease. A purposive sampling survey was conducted in four AEUs of Kerala viz., Southern Central Laterites (SCL), Southern and Central Foothills (SCFH), Northern Laterites (NL), and Northern Foothills (NFH). Three locations from each of these four AEUs were surveyed and samples of diseased plants and rhizosphere soil were collected during August to November 2019. Rhizosphere soil of healthy plants were also collected from each location. The per cent disease incidence in different locations ranged from 20.40 to 56.60 per cent. The bacterial wilt pathogen was isolated from infected plants using tetrazolium chloride (TZC) agar and Koch’s postulates were proved by raising seedling in polybags and further confirmed by inoculating seedlings in protrays. The pathogen population in the rhizosphere ranged from 1.33 to 102.67 x 106 cfu g-1 soil. The 12 isolates of bacterial pathogen collected from these locations were purified and maintained in sterile water for further experiments. Different isolates exhibited considerable variation in colony size and pigmentation. All the isolates were gram negative and scanning electron microscopy (SEM) revealed that, the bacterial cells are rod shaped with different sizes such as 0.47 to 0.69 µm x 1.46 to 1.94 µm in the case of NL 2; 0.50 to 0.52 µm x 1.48 to 1.69 µm in case of NL 1 and 0.52 to 0.59 µm x 1.50 to 1.94 µm in case of SCL 2. The molecular characterization of the isolates by 16S rRNA sequencing revealed that there are five different bacterial wilt pathogens present in four different AEUs of Kerala. The isolate SCL 1 was identified as Klebsiella variicola; SCL 2, SCFH 1, SCFH 2, and NL 3 were identified as Klebsiella pneumoniae; NFH 3, NL 2, SCFH 3, and SCL 3 were identified as Enterobacter mori. Further, NFH 1 and NL 1 were identified as Klebsiella oxytoca and NFH 2 was identified as Enterobacter cloacae. Phylogenetic analysis revealed that, the different isolates clustered together on the basis of the genus and species to which it belongs. However, due to absence of barcode gaps between Klebsiella and Enterobacter spp., at genus level using 16S rRNA sequencing, specific primer based molecular characterization by sequencing housekeeping genes rpoB, gyrB and hsp60 was carried out. In-silico analysis of rpoB gene sequences confirmed the identity of 11 isolates. Interestingly, SCFH 1, SCFH 2, SCFH 3, SCL 1, SCL 3 and NFH 2 were identified as Enterobacter sp. Whereas, NL 3, NFH 1 and NFH 3 were identified as Klebsiella sp. Moreover, SCL 2 and NL 1 were identified as Pantoea sp. In-silico analysis of gyrB gene sequences confirmed the identity of eight isolates. SCFH 1, SCFH 2, SCFH 3, SCL 1, SCL 3 and NFH 2 were identified as Enterobacter sp. Moreover, SCL 2 and NL 1 were identified as Pantoea sp. Similarly, In-silico analysis of hsp60 gene sequences also confirmed the identity of eight isolates. NL 3, NFH 1 and NFH 3 were identified as Klebsiella sp. Further, SCL 1, SCFH 1, SCFH 2 and SCFH 3 isolates were identified as Enterobacter sp. and SCL 2 isolate was identified as Pantoea sp. Concatenated phylogenetic analysis of rpoB, gyrB and hsp60 gene analysis depicted the grouping of different genera viz., Klebsiella sp., Enterobacter sp. and Pantoea sp. separately in different clusters. However, there were shift between different genera viz., Klebsiella to Pantoea sp., Klebsiella to Enterobacter sp. and Enterobacter to Klebsiella sp. within Enterobacteriaceae family while identifying using 16SrRNA and rpoB, gyrB, hsp60 gene sequencing. This indicates close genetic relatedness of these genera. This also indicates considerable genetic variation among the isolates present in different locations of AEUs. The isolates of these bacterial wilt pathogens were tested for pathogenicity on the reported host plants viz., tomato, brinjal, chilli, banana, potato, ginger and mulberry. However, the isolates infected only tomato, brinjal and chilli but could not infect the other host plants tested. These isolates were tested for their ability to utilize disaccharides and hexose-alcohols too. The isolates belonging to the same genus and species showed almost similar carbohydrate utilization patterns. The physico-chemical and biological properties of the soil samples collected were studied using standard protocols. Statistical analysis using paired sample t-test revealed that, there was no significant difference in chemical properties viz., pH, organic carbon, available P, K, Ca, Fe and Mg content between rhizosphere soil of healthy and diseased plants. Similarly, there was no significant difference in the physical properties of rhizosphere soil of healthy and diseased plants within the same location except for core bulk density. However, there is statistically significant difference in population of actinomycetes and fluorescent pseudomonads in rhizosphere of healthy and diseased soil. Moreover, dehydrogenase and catalase activity also showed a statistically significant difference between healthy and diseased plant rhizosphere soil. Metagenomic studies revealed that, rhizosphere soil of diseased plants has very high pathogen population compared to that of the healthy plants. However, bacterial diversity and richness was less in rhizosphere soil of wilted plants while that of healthy plants. Moreover, the microbial community structure in healthy and sick soil also showed difference. Even though three bacterial genera (Klebsiella, Enterobacter, Ralstonia) causing bacterial wilt disease were found in the soil of bacterial wilt infected and healthy plant’s rhizosphere, the bacterial population of Klebsiella genus was only significantly higher in diseased rhizosphere soil (SCFH1D) as compared with that of (SCFH1H) rhizosphere soil of the healthy plant. This suggests that an increase in the relative abundance of phytopathogenic bacteria K. pneumoniae is the cause of bacterial wilt disease in chilli. These findings are also evidently supported by the molecular characterization results of the bacterial wilt pathogen (SCFH1 isolate) obtained from this location which is also identified as K. pneumoniae based on 16S rRNA gene sequencing. These studies also revealed that some beneficial microbes were present in the rhizosphere of healthy chilli plant, however, shifts may occur between the rhizosphere bacterial flora of bacterial wilt infected and healthy plant’s rhizosphere. To develop an integrated management strategy for bacterial wilt disease, field experiments were conducted in a bacterial wilt sick plot at the Department of Vegetable Science, CoA, Vellanikkara. The 12 different treatments in the field experiment included application of bleaching powder, soil test-based lime application, seedling root dip in streptocycline, drenching of copper hydroxide, drenching of copper oxychloride, seed treatment and seedling dip with Pseudomonas fluorescens, drenching of P. fluorescens, planting grafted plants of tomato and other integration treatments. The results of the study revealed that to manage the bacterial wilt of tomato, an integration of different treatments viz., application of bleaching powder (15 kg/ ha) + soil test- based lime application + streptocycline (@ 2g/10L) root dip + drenching of copper oxychloride @ 0.3 per cent is required. Efficacy of silver nanoparticles against bacterial wilt pathogen was tested under in-vitro conditions and it was found that there is no antibacterial property against bacterial wilt pathogens Klebsiella sp., Enterobacter sp. and Pantoea sp. The study revealed that in all 12 different locations surveyed in four selected agro-ecological units, Klebsiella sp., Enterobacter sp. and Pantoea sp. cause bacterial wilt of solanaceous vegetables. There is statistically significant difference in core bulk density and various biological properties of rhizosphere soil of healthy and diseased plants. Metagenomic analysis revealed that, the density and diversity of bacterial population in rhizosphere soil of healthy plants are much higher compared to that of diseased plants. To manage bacterial wilt of tomato, integration of application of bleaching powder (15 kg/ ha) + soil test-based lime application + streptocycline (@ 2g/10L) root dip + drenching of copper oxychloride @ 0.3 per cent was found suitable.Item Exploitation of cytoplasmic genetic male sterility (CGMS)system i chilli (Capsicum annuum L.)for the development of hybrids resistant to bacterial wilt(Department of Vegetable Science ,College of Agriculture, Vellanikkara, 2025-12-18) Malavika, O; Pradeepkumar, TChilli botanically known as Capsicum annuum (2n=24, Solanaceae) is an important vegetable crop extensively cultivated in tropical and subtropical parts of the world. However, the crop faces major challenges due to its small flower size which complicates hybrid seed production and susceptibility to various biotic stresses, both of which restrict its yield potential. Developing host plant resistance to bacterial wilt through heterosis breeding using cytoplasmic gentic male sterility system and identified resistance sources is an effective method to mitigate the biotic stress and to realise the yield potential in chilli. With this context the current study entitled as “Exploitation of Cytoplasmic Genetic Male Sterility (CGMS) System in Chilli (Capsicum annuum L.) for the development of hybrids resistant to bacterial wilt” was carried out at the department of vegetable science during May 2022-Novemener 2024. Initially 17 chilli genotypes comprised of seven male sterile, maintainer and 10 restorer lines were evaluated at department of Vegetable Science, College of Agriculture, Vellanikkara, during May 2022-23. The statistical analysis of various characters exhibited significant differences among lines and restorer lines. The analysis on pollen sterility revealed that four lines were completely sterile, two were partially sterile and one line was found partially fertile throughout growing period. Among the lines, the highest plant height was recorded in VI046838 (116.66 cm), while VI060627 exhibited earliness in terms of days to flowering (24.56 days), fruiting (28.46 days), and maturity (44.30 days). Superior mean performance for fruit traits such as fruit weight, fruit length, fruit wall thickness, and fruit width was observed in VI046838. The mean performance of the genotypes for yield and quality parameters revealed notable differences. Fruit set percentage and fruit- bearing period were highest in AVPP9907, with values of 73.77% and 119.96 days, respectively. The highest fruit yield per plant and number of seeds were recorded in VI060629. The highest value of ascorbic acid observed in AVPP9907 (213.82 mg/100 g). Significant differences were also noted for oleoresin content, with AVPP9907 recording the highest value (18.10 %). Under field screening for bacterial wilt incidence, three genotypes were resistant, three were susceptible, and VI046838 was moderately susceptible. However, none of the seven genotypes showed resistance under artificial inoculation. Out of 10 restorer lines, AVPP9905 was found to be superior for multiple traits, including, days to first flowering, fruit weight, fruit length, fruit-bearing duration, number of fruits per plant, and overall yield. Chivar-1 showed the highest ascorbic acid content (197.83 mg/100 g), while LC 217 recorded the highest oleoresin content (18.37%). Highest capsaicin content was observed in AVPP9703 (1.06%). Bacterial wilt screening among the restorer lines revealed that seven were resistant, two were susceptible, and one was moderately resistant. High genotypic and phenotypic coefficient of variation was observed for fruit per plant, fruit length, fruit set percentage, fruit bearing period, fruit width, fruit wall thickness. Path analysis of the characters revealed that fruits per plant, fruit set percentage, and fruit bearing had a direct positive effect on yield, Twenty-four hybrids were produced from a cross involving 3 × 8 (L × T) mating design as a part of the second experiment. In third experiment Parents and hybrids, along with the standard check Sierra were evaluated for various characters. The ANOVA for combining ability revelated presence of significant general combining ability for parents and specific combining ability for hybrids. Predominance of non additive gene action was evident for almost all the characters under study. Significant heterosis was observed for most of the quantitative traits. Yield and contributing characters showed significant difference among parents, hybrids and check. The hybrid AVPP0516 × Anugraha exhibited the highest mean fruits per plant (179.65) and yield per plant (822.12 g). AVPP0517 was the best combiner for fruits per plant, Among the testers significantly highest positive GCA for yield per plant was noticed in Ujwala. In lines, AVPP9907 had highest GCA for yield per plant. Among the testers, Ujwala had maximum GCA for yield per plant. Regarding biochemical characters, VI059328 recorded highest GCA for ascorbic acid, Chivar-1 for oleoresin and AVPP9703 had highest GCA for capsaicin content. AVPP0516 × Anugraha, AVPP0517 × VI059328, and AVPP9907 × Ujwala were best combiner for yield per plant. For fruit setting percentage, highest SCA was observed in the cross AVPP0516 × LC 217. Highest SCA for ascorbic acid was observed in the hybrid AVPP0516 × VI059328. AVPP0517 × Ujwala exhibited maximum SCA for oleoresin. On estimation of heterosis for number of fruits per plant, the hybrid AVPP0516×Anugraha recorded superiority in terms of relative heterosis (137.88%), heterobeltiosis (102.26%) and standard heterosis (212.71%). Artificial screening of hybrids revealed that AVPP0517 × VI059328 was highly resistant, with a disease incidence of 10.05%, followed by AVPP0516 × Anugraha (10.35%) and AVPP0517 × EC566920 (11.50%). The reported DNA marker for bacterial wilt resistance was validated using the parental lines and their hybrids. The primer CAMS 451 exhibited polymorphism, producing the expected band size of 220 bp for susceptible genotypes and 210 bp for resistant genotypes. All 24 hybrids tested positive for the marker and were confirmed to be heterozygous in nature. The study stands out for the successful development of three promising F1 hybrids, namely, AVPP0516 × Anugraha, AVPP0517 × VI059328, and AVPP9907 × Ujwala. Based on their mean performance, combining ability, and heterosis, they were recognized as potential genotypes for further evaluation and utilization in breeding programs. Exceptional standard heterosis and resistance to bacterial wilt were also observed in four hybrid, AVPP0516 × Ujwala, AVPP0517 × Anugraha, AVPP0517 × Ujwala, and AVPP9907 × Anugraha which are potential candidates for replicated and multilocational testing under bacterial wilt prone zones in Kerala.Item Horticultural mineral oil for pest management in Brinjal and Chilli(Department of Agricultural Entomology, College of Agriculture, Padannakkad, 2026-01-31) Sajay,S; Sreekumar, K MThe research programme entitled ‘Horticultural mineral oil for pest management in brinjal and chilli’ was aimed at elucidate the mode of action, phytotoxicity, safety to natural enemies, compatibility and pest management efficacy of horticultural mineral oil in brinjal and chilli. Safety evaluation of horticultural mineral oil was carried out in the Department of Agricultural Entomology, College of Agriculture, Padannakkad, during 2022-23 to evaluate phytotoxicity, toxicity to natural enemies and compatibility with biocontrol agents. Eleven concentrations of HMO (HMO at 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5% and 8%) and a control were applied on one month old seedlings of brinjal and chilli as part of phytotoxicity evaluation. For experimenting on natural enemies and biocontrol agents, six concentrations of HMO (HMO at 2%, 2.5%, 3%, 3.5%, 4% and 4.5%) and a control were evaluated. All the safety evaluation experiments were designed under CRD with three replications. Results showed that HMO up to 6.5% did not show any phytotoxic symptoms in both brinjal and chilli. But HMO at 7%, 7.5% and 8% resulted in phytotoxicity symptoms like necrotic patches, with the severity of these symptoms showing an increasing trend as the concentration increased. HMO at 2% is safest for Trichogramma chilonis, Bracon brevicornis, Cheilomenes sexmaculata and Chrysoperla sp. followed by 2.5% and 3%. HMO is not compatible with biocontrol agents such as Beauveria bassiana, Metarhizium anisopliae, Lecanicillium lecanii and Trichoderma asperellum. Laboratory evaluation of horticultural mineral oil against selected pests of brinjal and chilli was carried out in the Department of Agricultural Entomology, College of Agriculture, Padannakkad, during 2022-23 to evaluate the toxicity against grubs of epilachna beetle Henosepilachna vigintioctopunctata and caterpillar of shoot and fruit borer Leucinodes orbonalis. A net house experiment was conducted to find the repellency of chilli whitefly, Aleurothrixus trachoides. The test organisms were exposed to seven treatments including T1: HMO at 2%; T2: HMO at 2.5%; T3: HMO at 3%; T4: HMO at 3.5%; T5: HMO at 4%, T6: HMO at 4.5% and T7: control designed under CRD with three replications. From the results, HMO at 4.5% caused the highest significant mortality of Leucinodes orbonalis which was on par with 4% followed by 3.5%, and 3%. HMO at 4.5% caused the highest significant mortality of Henosepilachna vigintioctopunctata, which is on par with 4% followed by 3.5%, and 3%. HMO at 4.5% resulted in the highest significant repellency against Aleurothrixus trachoides, which was on par with 4% followed by 3.5%, and 3%. HMO at 3% was selected as the best concentration based on the results of safety evaluation experiments and laboratory evaluation of selected pests. Field study was carried out using RBD with eight treatments and three replications on brinjal and chilli from November 2023 to April 2024 at the Instructional farm of College of Agriculture, Padannakkad. Treatments applied in brinjal were: T1: HMO at 3 %; T2: HMO at 3 % + azadirachtin formulation at 1 % 1ml/L; T3: azadirachtin formulation at 1 % 2ml/L; T4: HMO at 3 % + thiamethoxam 25WG 0.1g/L, T5: thiamethoxam 25WG 0.2g/L, T6: HMO at 3 % + chlorantraniliprole 18.5SC 0.15ml/L, T7: chlorantraniliprole 18.5SC 0.3ml/L and T8: untreated control and treatments applied in chilli were T1: HMO at 3 %, T2: HMO at 3 % + azadirachtin formulation at 1 % 1ml/L, T3: azadirachtin formulation at 1% 2ml/L, T4: HMO at 3 % + thiamethoxam 25WG 0.2g/L, T5: thiamethoxam 25WG 0.4g/L, T6: HMO at 3 % + fenpyroximate 5EC 0.3ml/L, T7: fenpyroximate 5EC 0.6ml/L and T8: untreated control. Observations on pest population were made a day prior to spraying and post treatment population at 1,3,5,7 and 14 DAT while damage symptoms were observed at 7 and 14 DAT. Results indicated that, HMO at 3% in combination with half the recommended dose of chemical pesticides (azadirachtin formulation 1 % 1ml/L, thiamethoxam 25WG 0.1g/L, and chlorantraniliprole 18.5SC 0.15ml/L) yielded the same significant results when compared with the full recommended dose (azadirachtin formulation 1% 2ml/L, thiamethoxam 25WG 0.2g/L and chlorantraniliprole 18.5SC 0.3ml/L) against epilachna beetle Henosepilachna vigintioctopunctata in brinjal. HMO at 3% in combination with chlorantraniliprole 18.5SC 0.15ml/L (half the recommended dose) and chlorantraniliprole 18.5SC 0.3ml/L (full recommended dose) achieved the highest reduction in the damaged shoots and fruits caused by the attack of shoot and fruit borer Leucinodes orbonalis in brinjal. HMO at 3 % in combination with thiamethoxam 25WG 0.1g/L (half the recommended dose) was superior among all other treatments against leaf hopper Amrasca biguttula biguttula in brinjal, which was on par with treatment thiamethoxam 25WG 0.2g/L (full recommended dose). HMO at 3% in combination with chlorantraniliprole 18.5SC 0.15ml/L recorded the highest marketable yield of brinjal, which was followed by chlorantraniliprole 18.5SC 0.3ml/L and HMO 3% in combination with azadirachtin formulation 1% 1ml/L. HMO at 3% in combination with fenpyroximate 5EC 0.3ml/L (half the recommended dose) achieved the best result, which was on par with fenpyroximate 5EC 0.6ml/L (full recommended dose) against chilli mite Polyphagotarsonemus latus in chilli. HMO 3% in combination with fenpyroximate 5EC 0.3ml/L and thiamethoxam 25WG 0.2g/L (half the recommended dose) resulted in highest success rate in controllling the thrips Thrips parvispinus population in chilli which was on par with fenpyroximate 5EC 0.6ml/L and thiamethoxam 25WG 0.4g/L (full recommended dose). HMO at 3 % in combination with thiamethoxam 25WG 0.2g/L (half the recommended dose) combination gave the best results among all other treatments against whitefly Aleurodicus disperses in chilli, which was on par with treatment thiamethoxam 25WG 0.4g/L (full recommended dose). HMO at 3% in combination with fenpyroximate 5EC 0.3ml/L recorded the highest marketable yield of chilli, which was followed by fenpyroximate 5EC 0.6ml/L. The scanning electron microscope (SEM) study conducted to find out the morphological changes in Myzus persicae and Spodoptera litura treated with HMO 3% indicated that the treatment changed the mouthpart morphology of Myzus persicae. SEM images also revealed that HMO 3% blocked the spiracles and damaged the trichomes, and produced significant changes in the structure of the cuticle of Spodoptera litura. A comparative study of the droplet spreading area of HMO, insecticides, and their combinations was designed under CRD with nine treatments and three replications. Treatments applied were: T1: HMO at 3 %; T2: HMO at 3 % + azadirachtin formulation at 1 % 1ml/L; T3: azadirachtin formulation at 1 % 2ml/L; T4: HMO at 3 % + thiamethoxam 25WG 0.1g/L, T5: thiamethoxam 25WG 0.2g/L, T6: HMO at 3 % + chlorantraniliprole 18.5SC 0.15ml/L, T7: chlorantraniliprole 18.5SC 0.3ml/L, T8: HMO at 3 % + fenpyroximate 5EC 0.3ml/L and T9: fenpyroximate 5EC 0.6ml/L. Results showed that, the droplets of the combinations having HMO at 3% and HMO at 3% alone spread more compared to chemical formulations which used alone. Since oil plus emulsifier in the HMO reduces surface tension and creates more stable dispersed droplets, when sprayed on leaves or other plant surfaces, the chemical combination with HMO can spread more effectively than with the chemical applied alone.Item User friendly reagents for rapid thermochemical processing of Biowaste(Department of Soil Science and Agricultural Chemistry, College of Agriculture, Vellayani, 2026-02-13) Greeshma, P R; Naveen, LenoA study entitled ‘User friendly reagents for rapid thermochemical processing of biowaste’ was carried out at the Department of Soil Science and Agricultural Chemistry, College of Agriculture, Vellayani during 2022-2024. The objective of the study was to standardize user friendly and easily available alternate reagents for rapid thermochemical processing of biowaste, study the nutrient release and retention characteristics of the resultant thermochemical organic fertilizer and test its crop suitability. The study included standardization of reagent protocol for production of thermochemical organic fertilizer (TOF) by testing of resultant products and selecting the best three alternate reagent combinations, characterization of those products, an incubation study, leaching study and field experiments using cowpea var. Kanakamony as test crop. Production of TOF was as per patented KAU rapid thermochemical processing technology developed by Sudharmaidevi et al. (2017). The acid-alkali components were substituted with reagents such as acetic acid (5% and 10%), calcium hydroxide (5%, 10%, 15%), and calcium oxide (10%, 15%). All formulations were neutral to slightly alkaline with TOC content ranging from 33.94% to 38.21%. Phytotoxicity assessment based on the germination index (GI) of cowpea, bhindi, and amaranthus indicated values exceeding 60% for TOFs prepared from 5 % Acetic acid + 10 % Ca(OH)2 [TA5CH10], 5 % Acetic acid + 15% Ca(OH)2 [TA5CH15], 10 % Acetic acid + 5 % Ca(OH)2 [TA10CH5], 10 % Acetic acid + 10 % Ca(OH)2 [TA10CH10], and the patented TOF (P), suggesting their safety and non-toxic nature for seed germination and seedling growth. In pot culture studies, TOF (P) recorded the highest yield (68.70 g/plant), which was statistically on par with TA10CH10 (65.83 g/plant). Number of fruits per plant, pod length and pod weight were statistically comparable for both TOF (P) and TA10CH10. This was followed by TA5CH10 which was statistically similar to that of TOF (P) in plant height, length and weight of pod. This was followed by TA5CH15. So TA10CH10, TA5CH10 and TA5CH15 were selected for further analysis. Characterization of these three selected TOFs showed physical and chemical properties meeting FCO standards. Among them, TA10CH10 recorded the highest levels of total N (2.08%), K (1.42%), Mg (0.71%), S (301.33 mg/kg), and all micronutrients. These findings emphasized the superior nutrient profile of TA10CH10 formulation. The incubation study for 90 days revealed the nutrient release pattern of organic fertilizers. The 100 and 75 per cent of TOF from the three selected reagent combinations, and TOF (P), FYM as per POP were incubated (17.85 g per container pot). TOF was applied in N equivalence of FYM (8.9 g per container pot for 100 % TOF and 6.69 g per container pot for 75 %TOF). Soil alone was kept as control. Soil pH in all treatments recorded a decreasing trend up to 60 days. EC of the soil was found increasing till 60 days followed by a slight decrease till 90 days. Soil organic carbon content and available N content was found increasing up to 30 days followed by a decrease till 90 days. Available P content was found to be higher in treatment where FYM was applied and an increasing trend was observed up to 30 days followed by a decline till 60 days and then an increase. Available K was found increasing till 60 days followed by a decrease and was recorded highest for TOF (P). Soil OC, available N, Mg, S and micronutrients were highest from TOF (P) and was significantly on par with TA10CH10. In the leaching study, sieved samples (40 g) were mixed with 800 mL distilled water and filtered samples were analyzed at different time intervals. Ammoniacal N increased, while total and nitrate N decreased. Leaching of other macronutrients increased over time. Micronutrients exhibited varying patterns with majority following an increasing trend. Overall, TOF (P) and TA10CH10 had the highest nutrient release, indicating greater leaching potential. Field trials showed that 100% TOF (P) + STBR (T5) recorded the highest cowpea yield in Rabi (33.48 g/plant) and Kharif (34.56 g/plant). STBR+ TA10CH10 (T4) was statistically on par with T5 in both seasons (31.86 g/plant and 31.45 g/plant). Plant height, number of nodules, number, length and weight of pod were found statistically similar between T5 and T4. Available nutrient content in post-harvest soil, whole plant analysis and crude protein content of T4 was found statistically on par with T5. The study concluded that application of TA10CH10 (TOF prepared from 10 % acetic acid and 10 % calcium hydroxide) at 10 t ha⁻¹, in combination with soil test based fertilizer recommendation (STBR), proved to be best for cowpea var. Kanakamony compared to both FYM + STBR and TOF formulations prepared using other alternate reagent combinations. Overall, TOF prepared from 10 % acetic acid and 10 % calcium hydroxide (TA10CH10) emerged as a promising, user-friendly, and sustainable alternative to the patented TOF—matching its nutrient efficiency and yield benefits while being easily produced from biowaste using vinegar and lime, making it ideal for small-scale household adoption.Item Microbiome complementation from Piper colubrinum for Foot rot disease (Phytophthora capsici) management in black pepper (Piper nigrum L.)(Department of Microbiology, College of Agriculture, Vellayani, 2026-02-16) Abarna, R; Anith,K NThe study entitled “Microbiome complementation from Piper colubrinum for foot rot disease (Phytophthora capsici) management in black pepper (Piper nigrum L.)” was carried out at the Department of Microbiology, College of Agriculture, Vellayani, Thiruvananthapuram. The research aimed to isolate and characterize culturable, and analyze the unculturable microbiome of the wild pepper; Piper colubrinum, and the cultivated Piper nigrum, and evaluation of a culturable microbial consortium or synthetic microbiome with special emphasis on suppression of foot rot disease of black pepper. Characterization of the microbiome of three Piper plant types; Piper colubrinum (wild; foot rot resistant), healthy Piper nigrum plants in a Phytophthora capsici affected field, and diseased Piper nigrum affected by foot rot/quick wilt was done from rhizospheric soil and root samples collected from Ambalavayal, Wayanad, Kerala. For each plant type, three soil and three root samples were pooled into single replicate. In the 16S rDNA metagenomic analysis, operational taxonomic units (OTUs) were mainly assigned to five dominant bacterial phyla: Proteobacteria (34.74%), Firmicutes (27.41%), Acidobacteriota (9.33%), Actinobacteriota (7.58%), and Bacteroidota (6.49%), with the remainder distributed among several other phyla. Bacillus was the only genus present across all samples, a finding supported by culture dependent assays, and was most abundant in Piper colubrinum samples. Bryobacter was also widely distributed, except in Piper nigrum roots. In 18S rDNA ITS metagenomic analysis, fungal communities were dominated by Ascomycota (83.73%), followed by Basidiomycota (8.55%), Rozellomycota (3.35%), Mortierellomycota (3.31%), and Mucoromycota (1.25%). Notably, Trichoderma asperellum, which exhibited the strongest anti-oomycete activity in both in vivo and in vitro assays, was detected exclusively in P. colubrinum and absent in P. nigrum. Rarefaction curve analysis revealed greater diversity in soil samples particularly of P. nigrum than in root samples. Comparative bacterial analysis (Venn diagram) showed the highest number of unique features in PNDS (Piper nigrum diseased soil), followed by PNHS (Piper nigrum healthy soil), PNDR (Piper nigrum diseased root), PNHR (Piper nigrum healthy root), PCHS (Piper colubrinum healthy soil), and PCHR (Piper colubrinum healthy root). For fungi, the most unique features were found in PNDS, followed by PCHS, with only eight features shared among all six groups. In the present study, fifty-one bacterial isolates and eleven fungal endophytes of Piper nigrum and twenty-two bacterial isolates and three fungal endophytes of Piper colubrinum were isolated and purified. A total of seventy-three bacterial isolates and eleven fungal endophytes from Piper species were initially evaluated for their antagonistic activity through dual culture plate assay, serving as the preliminary screening step for identifying efficient culturable microbiome candidates. From this screening, twenty bacterial isolates and seven fungal endophytes were shortlisted for further assessment using dual culture plate assay, detached leaf assay, and cut stem assay. Subsequently, their plant growth-promoting traits such as ability to grow in nitrogen-free medium, phosphorus and zinc solubilization, production of phytohormones such as indole-3-acetic acid (IAA) and gibberellic acid (GA), siderophores and hydrogen cyanide (HCN) production, ACC deaminase activity, ammonia production, and synthesis of both volatile and non-volatile organic compounds were analyzed. Seven bacterial isolates (PNS14, PNS17, PNRS3, PNRO6, PCRO5, PCRS1, PCRS5) and three fungal endophytes (PCLF1, PNRF1, PCSF2) that possessed antagonistic activity together with plant growth-promoting attributes were subjected to identification through morphological, biochemical, and molecular methods. These included PNS14 (Bacillus sp.), PNS17 (B. amyloliquefaciens), PNRS3 (B. drentensis), PNRO6 (B. subtilis), PCRO5 (B. velezensis), PCRS1 (B. subtilis), and PCRS5 (Bacillus sp.), as determined by 16S rRNA sequencing. Among the fungal endophytes identified using ITS sequencing, PCLF1 (Trichoderma asperellum) alone was selected for in vivo studies, as other species PNRF1 (Rhizoctonia solani), and PCSF2 (Fusarium fujikuroi) were supposed to be plant pathogenic, and therefore excluded from further experiments. For consortium preparation, three Piper colubrinum bacterial isolates (PCRO5, PCRS1, PCRS5) were combined to form Piper colubrinum consortium PCCB, while four Piper nigrum bacterial isolates (PNS14, PNS17, PNRS3, PNRO6) were combined to form Piper nigrum consortium PNCB. All bacterial isolates (PCCB + PNCB) along with the fungal endophyte T. asperellum (PCLF1) were used as different treatments for in vivo assays. A polybag experiment was carried out with consortia of isolates from Piper species to assess their biocontrol potential and ability to promote plant growth in black pepper. Healthy two-node cuttings treated with the consortia from Piper spp. exhibited notable improvements in biometric traits, including number of leaves, number of branches, plant height, fresh weight and dry matter accumulation, when compared to untreated controls. Moreover, growth parameters were significantly higher in cuttings inoculated with the consortium than in those treated with indole-3-butyric acid (IBA) and control. A foliar infection assay was conducted to assess the disease-suppressing potential plant associated microbes of Piper spp. using two sets of treatments: set 1, seven individual bacterial isolates were applied separately as treatments, while set 2 consisted of consortial treatments (PCCB, PCCF, PNCB, and PCCB+PNCB). Lesion development was monitored from one to seven days after inoculation (DAI) with the pathogen. In Set 1, the lowest disease index was recorded in plants treated with Bacillus sp. PCRS5 (0.08), followed by Bacillus subtilis PNRO6 (0.10). In Set 2, PCCF showed the lowest disease index (0.08), achieving 73.2 % disease suppression compared to the pathogen control, followed by PCCB and PNCB (0.11 each). Furthermore, treated plants of set 1 and set 2 were evaluated for defense-related enzyme activities, including peroxidase, polyphenol oxidase, phenylalanine ammonia lyase, superoxide dismutase (SOD), catalase, and β-1,3-glucanase. The results showed that, following P. capsici infection, these treated plants exhibited a statistically significant (p < 0.05) increase in enzyme activity compared to their levels prior to pathogen inoculation. Plants in the treatments PCCB and PCF exhibited higher survival rates (70.2 % and 73.2 %, respectively) compared to the inoculated control, when the pathogen was administered through soil. The development of a liquid formulation of the Piper spp. consortia, coupled with an analysis of bacterial and fungal population dynamics, revealed a pronounced capacity for rapid proliferation, occurring within minutes to hours. Over the course of a six-month experimental period, population stability was consistently maintained, in formulations supplemented with 2 % glycerol, 2 % PVP and 15 mM trehalose for bacteria, while fungal populations were stably maintained in talc formulation, with the colony-forming unit (cfu/mL) counts of both bacteria and fungus being recorded. The findings of this study highlight the potential of the Piper colubrinum fungal isolate Trichoderma asperellum PCLF1, along with the Piper nigrum isolates PNRO6 (Bacillus subtilis) and PCRS5 (Bacillus sp.), in promoting plant growth, providing disease protection, and enhancing defense responses in black pepper. The microbiome profiles of healthy and diseased Piper nigrum, and Piper colubrinum emphasize the suitability of these isolates for effective management of P. capsici induced foot rot disease in black pepper.Item Potential site selection for water harvesting in a micro watershed with future water balance perspectives(Department of Soil and Water Conservation Engineering Kelappaji College of Agricultural Engineering and food technology, Tavanur, 2026-02-11) Aravind, P; Sindhu, P VWorld is facing acute water scarcity owing to growing freshwater demand, rainfall aberrations and miserable management of water resources. Rapid urbanization, climate change and unplanned land use have significantly altered the hydrological cycle, leading to unpredictable rainfall, declining groundwater levels, and hydrological extremes. Valanchery micro-watershed of Bharathapuzha river basin faces acute seasonal water scarcity due to its abrupt topography and higher runoff. In this regard, the study was carried out to identify the potential water harvesting sites with the scientific integration of future LULC, climatic projections, hydrological modeling and machine learning for the Valanchery micro watershed in Bharathapuzha river basin of Kerala. LULC classification was carried out for the years 2015, 2020 and 2023 using Google Earth Engine and Random Forest classification techniques. Overall accuracy of 0.84-0.89 and Kappa coefficient of 0.79 to 0.87 showed a satisfactory classification. LULC analysis revealed an increase in the urban and plantation and a decrease in waterbody, paddy and barren land. The land use of 2025 and 2030 was predicted using MOLUSCE plugin of QGIS software after validation of 2023 predicted land use with the actual land use map of 2023. Kappa coefficient of 0.79 was achieved during the land use prediction process and the percent of correctness was around 78.65 %. Future LULC predictions showed an increase of 1.18 km2 in urban and a reduction in paddy and plantation land use. SWAT hydrological modeling was carried out, for the present period and after satisfactory calibration (2021-2024) and validation (2024-2025), it was used for simulating the future water balance. Water balance analysis indicated that around 40% of rainfall exits as surface runoff, while groundwater recharge contribution was around 15%, which may be due to limited infiltration which emphasize the need for improved groundwater recharge structures. Results of future hydrological simulations revealed increasing monsoon intensities, rising temperatures (up to +1.5°C), and altered surface runoff–baseflow dynamics, particularly under SSP585 scenario. Compromise programming (CP) technique was used to identify the best model based on the observed rainfall and temperature data. Based on the CP results, it was found that the CESM2-WACCM was the best model for rainfall and the EARTH model was identified as the best model for the maximum and minimum temperature. Bias correction was carried out using the linear scaling method for the temperature and power transformation method for the precipitation data. Bias corrected rainfall and temperature data was incorporated into the calibrated SWAT model, which successfully simulated the water balance components for the present as well as for the near future (2025 and 2030). Future water balance, shows an increased rainfall and temperatures, leading to the hydrological shifts from infiltration-driven to runoff-dominated by 2030. Machine Learning models were applied based on IMSD guidelines to identify potential sites for water harvesting based on the future water demand of 2025 and 2030. Machine learning algorithms combined with IMSD criteria were evaluated using ROC-AUC metrics. The XG Boost achieved the highest accuracy with an AUC value of 0.92 followed by random forest with an AUC value of 0.75 and so XG Boost method was used to locate suitable sites farm ponds, percolation ponds, and check dams. Results indicated that larger structure density and spatially distributed water harvesting sites under SSP126, while SSP585 required higher density and strategically placed structures to mitigate intensified surface flow and seasonal droughts. The study establishes a practical framework that couples remote sensing, hydrological modelling, climate science, and ML-based site suitability, offering a robust decision-support tool for policy makers and watershed development agencies. This research contributes a scientifically validated and operationally feasible approach for climate-resilient watershed planning. Implementing the conservation measures in the identified sites will support water security, reduce flood risk in downstream areas and would enhance agricultural water availability in dry seasons, and will help to improve the long-term hydrological balance of the micro-watershed.Item Potential site selection for water harvesting in a micro watershed with future water balance perspectives(Department of Soil and Water Conservation Engineering, Kelappaji College of Agricultural Engineering and food technology, Tavanur, 2026-02-11) Aravind, P; Anu, VarugheseWorld is facing acute water scarcity owing to growing freshwater demand, rainfall aberrations and miserable management of water resources. Rapid urbanization, climate change and unplanned land use have significantly altered the hydrological cycle, leading to unpredictable rainfall, declining groundwater levels, and hydrological extremes. Valanchery micro-watershed of Bharathapuzha river basin faces acute seasonal water scarcity due to its abrupt topography and higher runoff. In this regard, the study was carried out to identify the potential water harvesting sites with the scientific integration of future LULC, climatic projections, hydrological modeling and machine learning for the Valanchery micro watershed in Bharathapuzha river basin of Kerala. LULC classification was carried out for the years 2015, 2020 and 2023 using Google Earth Engine and Random Forest classification techniques. Overall accuracy of 0.84-0.89 and Kappa coefficient of 0.79 to 0.87 showed a satisfactory classification. LULC analysis revealed an increase in the urban and plantation and a decrease in waterbody, paddy and barren land. The land use of 2025 and 2030 was predicted using MOLUSCE plugin of QGIS software after validation of 2023 predicted land use with the actual land use map of 2023. Kappa coefficient of 0.79 was achieved during the land use prediction process and the percent of correctness was around 78.65 %. Future LULC predictions showed an increase of 1.18 km2 in urban and a reduction in paddy and plantation land use. SWAT hydrological modeling was carried out, for the present period and after satisfactory calibration (2021-2024) and validation (2024-2025), it was used for simulating the future water balance. Water balance analysis indicated that around 40% of rainfall exits as surface runoff, while groundwater recharge contribution was around 15%, which may be due to limited infiltration which emphasize the need for improved groundwater recharge structures. Results of future hydrological simulations revealed increasing monsoon intensities, rising temperatures (up to +1.5°C), and altered surface runoff–baseflow dynamics, particularly under SSP585 scenario. Compromise programming (CP) technique was used to identify the best model based on the observed rainfall and temperature data. Based on the CP results, it was found that the CESM2-WACCM was the best model for rainfall and the EARTH model was identified as the best model for the maximum and minimum temperature. Bias correction was carried out using the linear scaling method for the temperature and power transformation method for the precipitation data. Bias corrected rainfall and temperature data was incorporated into the calibrated SWAT model, which successfully simulated the water balance components for the present as well as for the near future (2025 and 2030). Future water balance, shows an increased rainfall and temperatures, leading to the hydrological shifts from infiltration-driven to runoff-dominated by 2030. Machine Learning models were applied based on IMSD guidelines to identify potential sites for water harvesting based on the future water demand of 2025 and 2030. Machine learning algorithms combined with IMSD criteria were evaluated using ROC-AUC metrics. The XG Boost achieved the highest accuracy with an AUC value of 0.92 followed by random forest with an AUC value of 0.75 and so XG Boost method was used to locate suitable sites farm ponds, percolation ponds, and check dams. Results indicated that larger structure density and spatially distributed water harvesting sites under SSP126, while SSP585 required higher density and strategically placed structures to mitigate intensified surface flow and seasonal droughts. The study establishes a practical framework that couples remote sensing, hydrological modelling, climate science, and ML-based site suitability, offering a robust decision-support tool for policy makers and watershed development agencies. This research contributes a scientifically validated and operationally feasible approach for climate-resilient watershed planning. Implementing the conservation measures in the identified sites will support water security, reduce flood risk in downstream areas and would enhance agricultural water availability in dry seasons, and will help to improve the long-term hydrological balance of the micro-watershed.Item Combining yield and leaf curl-mosaic complex virus tolerance in Chilli(Department of Genetics and Plant breeding, College of Agriculture, Vellayani, 2026-02-16) Varsha, P Vengilat; Seeja, GChilli (Capsicum spp.) is a widely cultivated crop in India, valued for its economic and culinary significance as vegetables, spices, and in industrial applications such as the production of oleoresins. It is grown for its fresh green and dried red fruits. India is one of the largest producers, consumers, and exporters of chilli, with major growing states including Andhra Pradesh, Telangana, Karnataka, Tamil Nadu, and Maharashtra. Despite its significance, chilli production in India faces several challenges, primarily due to the prevalence of diseases that adversely affect yield and quality. Chilli Leaf Curl Mosaic Complex (CLCMC) is one of the most severe viral diseases affecting chilli (Capsicum spp.) production, especially in tropical and subtropical regions like India. This disease results from a synergistic interaction between multiple viruses, primarily the Chilli leaf curl virus (ChiLCV), a member of the Begomovirus genus, often in association with Tomato leaf curl virus (ToLCV), and sometimes with other satellite molecules or helper components. In severe cases of disease infection, up to 100 percent loss in marketable yield has been reported. Hence, a study entitled ‘combining yield and leaf curl mosaic complex virus tolerance in chilli’ was carried out in the Department of Genetics and Plant Breeding, College of Agriculture, Vellayani during 2020-2024, with the objective of identification of superior cross combinations for leaf curl-mosaic complex virus tolerance, yield and yield attributes in chilli. In the first experiment, a total of sixty-four genotypes, collected from National Bureau of Plant Genetic Resources, Kerala Agricultural University and farmers’ field were screened for natural incidence of chilli-leaf curl-mosaic complex disease in cubic lattice design during the summer of 2023. The highest mean yield per plant (g) was shown by Paal Mulaku (Kumarichanda Local). The genotypes showed significant differences for all the traits under study indicating the presence of substantial genetic variability. These sixty-four genotypes were classified into nine clusters based on Euclidean2 method. A modified disease scoring scale of zero to five was developed for leaf-curl mosaic complex disease by combining the disease scores of chilli leaf curl and cucumber mosaic virus. Disease incidence and severity calculated as vulnerability index were calculated based on disease scoring. The genotypes were classified into immune, tolerant, highly resistant, resistant, susceptible and highly susceptible based on the vulnerability index. Ten genotypes showed immune reaction, while twenty showed tolerance to CLCMC. Hence, a total of thirty genotypes were selected for artificial screening against CLCMC in pot culture experiment. Graft transmission technique was adopted for artificial inoculation during experiment II. The viruses were transmitted from the diseased Vellayani Athulya, used as scion, to the genotypes selected from experiment I, used as rootstocks. Six out of the eleven genotypes that were found to be immune to the disease complex belonged to the top ten high yielding genotypes. Hence, genotypes namely, Venjaramood Local II, KD 2, Unda Chilli (Vadakara Local), AP-1, IC-208580 and IC-570376 were selected as parents for the hybridization program to combine high yielding character and tolerance to chilli leaf curl-mosaic complex disease. Of these the genotype IC-208580 belongs to C.frutescens and Venjaramood Local II, KD-2, Unda Chilli (Vadakara Local), AP-1and IC-570376 belong to C.annuum. The virus titre was estimated based on virus specific primers. No amplification was obtained for the six genotypes when chilli leaf curl virus coat protein specific primer, CLA 7F/CLA 8R was used for the titer estimation while three genotypes showed amplification when degenerate Deng primer was used. This may be attributed to the ability of genotypes to suppress the disease symptom expression. The molecular markers reported to be linked to the diseases were validated in the second part of experiment II. SSR Markers namely PAU-LC-343-1, CA 516044, AVRDC MD*850 and AVRDC-PP188 were reported to be linked to the chilli leaf curl disease resistant gene. SSR Marker HpmsE003 was reported to be linked to cucumber mosaic virus resistance gene. All the markers showed polymorphism between resistant and susceptible genotypes. SSR marker HpmsE003 linked to cmr1 locus, also showed polymorphism between resistant genotypes and susceptible check. The six genotypes were hybridized in a diallel fashion during experiment III. A total of thirty cross combinations were made, with six parents, of which ten cross combinations were interspecific crosses (Capsicum frutescens X Capsicum annuum). Only three of the ten interspecific crosses germinated. Reciprocal crosses: Capsicum annuum X Capsicum frutescens did not germinate. Effectively the total number of crosses was reduced to twenty three. During experiment IV, F1 plants were raised along with their parents for evaluation of yield traits under field conditions with the application of plant protection chemicals and without application of plant protection chemicals for natural disease incidence screening. Vellayani Athulya was used as check variety. All the characters showed non-additive gene action except for fruit girth in the experiment with the application of plant protection chemicals. Highly significant gca effects for fruit weight, number of fruits per plant and yield were shown by the parents Venjaramoodu Local II (P1), IC-570376 (P6) and KD-2 (P2) respectively. High sca effect for number of fruits per plant and fruit yield per plant was shown by the cross combinations, AP-1(P4) x Unda Chilli (Vadakara Local) (P3) and KD-2 (P2) x Venjaramoodu Local II (P1) respectively. Based on mean performance and heterosis the cross combination IC 208580 (P5) x IC-570376 (P6) and Venjaramoodu Local II (P1) x KD-2 (P2) were found superior. Five cross combinations were identified for high yielding character under plant protected conditions while eight cross combinations showed tolerance to leaf curl- mosaic complex disease under natural field conditions. A total of five cross combinations were found to be superior to leaf curl mosaic disease tolerance and yield namely: IC-208580 (P5) x IC-570376 (P6), Venjaramoodu Local II (P1) x KD-2 (P2), KD-2 (P2) x Venjaramoodu Local II (P1), IC-570376 (P6) x KD-2 (P2) and KD-2 (P2) x IC-570376 (P6). The superior cross combinations selected for high yielding capacity along with tolerance to leaf curl-mosaic complex disease can be used in future breeding programs to develop superior high yielding and leaf curl mosaic complex tolerant chilli varieties.Item Process optimisation and quality evaluation of meal replacers for adults(Department of Community Science, College of Agriculture, Vellanikkara, Thrissur, 2026-04-04) Athira Raj; Suman, K TMaintaining balanced nutrition has become increasingly challenging for adults as a result of rapid urbanisation, changing lifestyles, and limited time available for meal preparation and consumption. This highlights the importance of developing convenient, affordable, and nutritionally balanced meal alternatives as a significant area of focus in current nutrition research. The present study on “Process optimisation and quality evaluation of meal replacers for adults” was proposed to standardise meal replacers using rice, ragi and other functional ingredients, and to evaluate the quality attributes of the developed products. The study also envisaged to incorporate the developed products with resistant starch and to prepare reconstituted probiotic products from meal replacers, and their quality evaluation. Chocolate and vanilla flavoured meal replacers were standardised in 4 sets each with 10 treatments using germinated and ungerminated rice (40-60%) and ragi (0-20%) flours, banana flour and jackfruit flour either alone or in combination (0-15%) as major ingredients. Soy protein isolate (5%), skimmed milk powder (15%) and other ingredients (defatted peanut flour and rice bran flour) at 5% level were added in all treatments. After combining all ingredients at required level for different treatments, roasting as well as gelatinisation followed by cabinet drying were employed for the preparation of meal replacers. For organoleptic evaluation, 100g of standardised meal replacers were reconstituted with 300 mL of luke warm water before serving. Among different treatments from eight sets, T5 (50% rice, 10% ragi, 15% jackfruit flour, 15% skimmed milk powder, 5% soy protein isolate, and 5% other ingredients) was selected as the best combination with mean scores above 8.0 for all sensory parameters. The standardised meal replacers were adequate in carbohydrates (62.09 - 66.20%), high in protein (22.88 - 25.28%), low in fat (1.84 - 2.32%) with a crude fibre content of 3.08 to 3.46 per cent. The energy value of meal replacers ranged from 365.85 to 376.59 Kcal/100g, which aligned with the recommended calorie of 200 to 400 Kcal specified for meal replacers by Codex standards. The glycaemic index of the formulations was found to be low (43.44 to 51.61). The minerals were highest in meal replacers prepared with ungerminated flours, whereas B vitamins were found to be highest in germinated flour based ones. Germination and gelatinisation significantly improved in vitro digestibility of starch and protein and in vitro availability of minerals. Gelatinisation improved the reconstitution properties, providing better consistency and mouth feel to the products. During six months of storage, the product was highly shelf stable under ambient condition with slight reduction in physico chemical, nutritional and organoleptic qualities. The selected meal replacers were incorporated with 25% of rice based resistant starch (RS3), replacing the rice portion, which increased the resistant starch content by 4 fold in all formulations. It had a positive impact on the consistency and mouth feel of the products. The inclusion of RS3 significantly increased the total starch (44.84 52.40g/100g) and crude fibre (3.93-4.31g/100g) with significant reduction in starch digestibility (56.13-79.27%) and glycaemic index (37.50-46.75). Wide variation in protein, fat, vitamins and mineral contents was not noticed in meal replacers with RS3 addition. The RS3 incorporated meal replacers were also highly shelf stable upto six months of storage. Resistant starch incorporated meal replacers were successfully utilised for developing probiotic set yoghurt and fermented shake using direct vat set (DVS) cultures of Lactobacillus acidophilus (LA-5). Set yoghurt standardised with 10% meal replacer slurry + 90% milk was found be acceptable based on sensory parameters. Optimised growth conditions for LA-5 in set yoghurt was 50g substrate, 4h incubation and 0.1g inoculum concentration with probiotic viability of 96 to 112×109 CFU/mL. Probiotic fermented shakes were standardised in milk using 40 % of roasted and 30% of gelatinised meal replacers as different sets. The optimised fermentation parameters for shake were 50g substrate, 15min. incubation with 0.5g inoculum, supporting high probiotic viability (99 to109×10⁹ CFU/mL). Probiotic set yoghurts and fermented shakes exhibited improved textural and nutritional profiles compared to their non-probiotic counterparts, characterised by higher viscosity, enhanced B vitamins, improved protein and starch digestibility (above 80%), with increased mineral availability ( above 70%) indicating superior nutrient assimilation. The products maintained microbial safety and strong probiotic viability of 83 to 101×10⁹ CFU/mL in set yoghurt and 90 to 102×10⁹ CFU/mL in fermented shakes for 15 days and 10 days of refrigeration respectively, confirming both functional stability and quality retention. Cost of production for meal replacers was found to be approximately Rs.50/100g. Probiotic incorporation doubled the production cost of yoghurt and fermented shake. The developed meal replacers are convenient, ready to use, and cost effective formulations that align well with the nutritional needs and lifestyle pattern of adults. The successful formulation of reconstituted products further demonstrated the meal replacers versatility, enhancing its potential for wider consumer acceptance and commercial application.Item Nutritional Status of menopausal women and formulation of functional foods(Department of Community Science, College of Agriculture, Vellanikkara, 2026-01-07) Anna, Reenu Shaji; Seeja, Thomachan PanjikkarnMenopause represents a natural and permanent cessation of menstruation, and generally occurs between 45 and 55 years of age. With the postmenopausal period extending to almost 30 years, representing one-third of a woman’s life, menopause has emerged as a significant public health concern. In this context, the present study aimed to assess the nutritional status of menopausal women and develop functional foods tailored to their needs. The study was conducted among 300 menopausal women aged 45–55 years, selected through stratified random sampling. Data was collected using a pre-tested interview schedule to obtain information on socioeconomic status, dietary patterns, menopausal history, and symptom severity. Socioeconomic status was assessed using the updated Kuppuswamy Scale (2024). The Menopause Rating Scale (MRS) was used to assess symptom intensity. Anthropometric measurements, body composition analysis, and dietary intake assessments (24-hour recall, food frequency and one-day food weighment) were conducted. Clinical and biochemical examinations were carried out in a subsample of 30 women by qualified medical practitioner. Results revealed that 72.33% of the participants were between 50–55 years, and 27.67% were between 45–50 years. The nuclear family system was predominant, and most women were engaged in sedentary or moderately active occupations. A large proportion (58.67%) reported health issues and 55.00% were on medication. However, regular exercise (36.67%) and adequate sleep (43.00%) were practiced by less than half. The most common comorbidities included diabetes mellitus, osteoporosis, and hypertension, while severe hair fall and fatigue were the most reported symptoms. Natural menopause was experienced by 84.00% of women, while 16.00% underwent surgically induced menopause; the latter was more common among women who had their first child birth at a later age. Women from lower socioeconomic groups experienced more severe symptoms. Menopausal symptoms spanned somatic, psychological, and urogenital domains. Among the somatic symptoms assessed, joint and muscular discomfort was the most common, reported by 89.33% of the women, followed by sleep problems (95.00%), heart discomfort like palpitations, breathing difficulty, variation in blood pressure (99.00%), and hot flushes (86.33%). Although hot flushes were widespread, they were mostly mild to moderate in severity, whereas joint and muscular discomfort showed a larger proportion of moderate to severe cases, indicating a greater impact on daily functioning. Only 1% of women reported an absence of heart discomfort such as palpitation, variation in blood pressure and breathing difficulty, while 5% did not experience sleep problems. These findings reveal that somatic symptoms were prevalent among nearly all participants. Anthropometric and body composition assessments revealed a high prevalence of overweight and obesity, with only 10.67% having normal BMI, 43.33% overweight, and 45.67% obese. Central obesity was evident in 63.67% with waist to hip ratio >0.85. High fat mass (25.00%), visceral fat (19.33%), and low skeletal muscle mass (31.33%) indicated a tendency toward sarcopenic obesity. Higher BMI, fat mass, and visceral fat were positively correlated with greater symptom severity, whereas lower bone and protein mass were associated with more severe menopausal manifestations. Clinically, hair thinning (33.33%), joint pain (43.33%), and hot flushes (23.33%) were predominant, with skin dryness and pigmentation more common among surgically induced menopausal women. Dietary assessment revealed that 97.33% were non vegetarians, though only 6.00% were aware of phytoestrogens and 3.00% recognised foods beneficial during menopause. 24-hour recall method revealed a marked imbalance in the intake of macronutrients and essential micronutrients. Protein intake was nearly adequate (92–93% of RDA), while fat intake exceeded the recommended levels (111–119% of RDA). In contrast, energy intake was inadequate, meeting only 56–74 per cent of the RDA. The intake of critical micronutrients such as calcium (17.41–18.09%), iron (21.44–21.48%) and zinc (21.81–22.12% of RDA) was alarmingly low, largely attributable to inadequate consumption of milk and milk products, green leafy vegetables and fruits. A similar dietary pattern was observed in the subsample of 30 menopausal women assessed using the one-day food weighment method. Energy intake met only 59–79.50 per cent of the RDA, whereas protein and fat intakes were comparatively satisfactory, meeting 84.90–90.80 per cent and up to 107.20 per cent of the RDA, respectively. However, pronounced deficiencies in essential minerals persisted, with calcium intake ranging from 16.81–18.61 per cent and iron from 11.10–17.72 per cent of the RDA. The overall findings indicate a dietary pattern characterised by high intake of visible fats, animal protein sources and inadequate intake of mineral dense plant foods, thereby increasing the risk of osteoporosis, anaemia and compromised immunity during the menopausal transition. Based on the identified nutritional gaps, functional foods rich in fibre, unsaturated fatty acids, calcium, vitamin D, and phytoestrogens were developed to support menopausal health. Two products, nutrimix powder and crackers were formulated and evaluated. The nutrimix powder, optimised with 20% defatted soy flour, 30% jackfruit seed flour, 30% malted ragi flour, 10% quinoa seed flour, 5% chia seed flour and 5% almond flour incorporation, showed excellent sensory acceptability by 20 judges. Physico-chemical analysis over four months of storage indicated a gradual increase in moisture (4.95% to 6.12%), with stable water absorption and solubility indices, confirming good storage stability. Similarly, composite crackers formulated with oats (50%), whole wheat (25%), defatted soy flour (5%) and 20% other ingredients demonstrated high organoleptic scores. During four months of storage, moisture increased, and energy declined, with minimal changes in other nutrients. The production cost per 100g was 66/- for nutrimix and 44/- for crackers, indicating affordability and feasibility for large scale preparation. The study highlights the nutritional vulnerability of menopausal women, characterised by poor diet quality, obesity, and micronutrient deficiencies, which collectively intensify menopausal symptoms. The developed nutrimix powder and composite crackers proved to be nutritionally rich, shelf stable, microbiologically safe, and organoleptically acceptable, demonstrating potential as functional foods to improve the nutritional well-being of menopausal women.