Phoenix phylogeny, and analysis of genetic variation in a diverse collection of date palm(Phoenix dactylifera) and related species

Date palm(Phoenix dactylifera),one of the most ancient crops,is grown commercially in>30 countries.Using whole plastome assemblies,phylogenetic analyses revealed that cultivated date palm accessions share the same clade with Phoenix sylvestris,Phoenix pusilla and Phoenix acaulis,which are native to the Indian subcontinent,and Phoenix caespitosa that is native to the Arabian Peninsula and the deserts of Somalia.Analysis of genetic diversity and genetic relationships among date palm accessions from 13 producing countries involved 195 date palm accessions that were genotyped at 19 microsatellite loci.Extensive genetic diversity was observed,with many accessions heterozygous for most markers in this clonally propagated crop.The average number of alleles per locus(42.1),expected heterozygosity(0.8),observed heterozygosity(0.47)and fixation indices(FST=0.42)demonstrated substantial genetic diversity and population structure.Iraqi accessions were found to have the richest allelic diversity,and the most private alleles.The model-based Bayesian method indicated that these accessions could be broadly divided into two structure groups,one group with predominantly African accessions and another predominantly Asian.Some germplasm,especially from Tunisia and Iraq,deviated from this generalization.Many accessions in the STRUCTURE-derived groups were found to be genetic admixtures,with gene flow between Asian and African groups.Indian and Pakistani date palms were found to be most closely related to North African germplasm.

Antimicrobial Profile of Lactic Acid Bacteria Isolated from Vegetables and Indigenous Fermented Foods of India against Clinical Pathogens Using Microdilution Method

In dairy and food industries lactic acid bacteria （LAB） have been used in form of starter culture that plays vital role in fermentation; as flavouring and texturizing or as preservative agents. There is increasing evidence that lactobacilli which inhabit the gastrointestinal tract develop antimicrobial activities and participate in the host＇s defence system[1]. During fermentation, most of the LAB produces a number of different compounds like organic acids, hydrogen peroxide, diacetyl, acetaldehyde, carbon dioxide, polysaccharides, and proteinaceous compounds called bacteriocins or bacteriocinogenic peptides.

Screening of antiproliferative activity mediated through apoptosis pathway in human non-small lung cancer A-549 cells by active compounds present in medicinal plants

Objective: To explore the antiproliferative activity and apoptosis in cells caused by active compounds present in plants using different techniques. Methods: We investigated the antiproliferative effects of methanolic extracts from different parts of seven plants on A-549(lung cancer) cells and primary cell culture(chick embryo fibroblast cells, as normal cells) using MTT assay and the potent plant was fractioned further. All these fractions were screened again for anti-proliferative activity. DNA fragmentation and DAPI staining were used to study apoptosis. Quantitative real-time was used to investigate the expression of apoptoticrelated genes. LC-MS and 1 H-NMR techniques were used to identify the active compounds present. EnzCheck caspase-3 assay kit was used to measure caspase-3 activity. Results: Methanolic extract of Vitex negundo(V. negundo) was selected as a potent fraction. Among all fractions screened, ethylacetate fraction of V. negundo was selected as the most potent antiproliferative fraction and phytochemical analysis of the extract revealed the presence of secondary metabolites. Ethaylacetate fraction of V. negundo was found to cause characteristic apoptotic morphological changes and generation of ROS in A-549 cells. Ethaylacetate fraction of V. negundo also induced apoptosis in A-549 which was supported by DNA fragmentation and DAPI staining. To investigate the molecular mechanism behind the cytotoxic effect of ethaylacetate fraction of V. negundo, quantitative real-time PCR was used to measure expression levels of p53, bax, bcl2, casp-3 and casp-9. Using LC-MS and 1 H-NMR techniques, cytotoxic compounds(luteolin and p-hydroxy benzoic acid) were identified which increased casp-3 activity in a dose and time-dependent manner in A-549 treated cells. Conclusions: It is concluded from the present study that V. negundo is capable of triggering growth-inhibitive and apoptosis effects in A-549 cells, signifying that V. negundo may possesses anti-lung cancer activity.

Integration of morpho-physico-biochemical traits with SSR and SRAP markers for characterization of castor genotypes of Indian origin

Castor(Ricinus communis L.)is an important tropical oilseed crop,whose oil has versatile,practical value,especially in industries.The present study aimed to estimate the nature and magnitude of variability in the castor germplasm concerning yield and its component traits and physico-biochemical characters.Seed yield per plant and oil content ranged from 80.90 g(ICS-165)to 248.30 g(RG-3216),and 34.7%(ICS-172)to 58.7%(JI-277),respectively.The iodine value of oil ranged from 76.36(JI-370)to 89.84(P2-135)with an average value of 83.02.The mean saponification value of oil was 182.24.The genotypic and phenotypic coefficients of variation were high for acid value,capsules on the main raceme,seed yield per plant,and total length of the main raceme.A positive association of porosity,average unit volume,and total length of the main raceme with seed yield per plant showed that these characters might be directly attributed to seed yield improvement.By Manhattan distances,the 30 genotypes were grouped into 3 clusters.Their genetic diversity was elucidated using SSR and SRAP markers.SRAP marker produced higher mean number of total bands(5.71),polymorphic bands(4.57),percentage polymorphism(83.10%),PIC(1.72),RP(5.90),mean RP(1.02),MR(5.71),EMR(4.57)and MI(1.44)values when compared to SSR(2.89,2.11,79.63%,0.61,1.90,0.72,2.89,2.11 and 0.49,respectively)marker.The highest genetic distance(0.77)was between 48-1 and JI-370,which indicated that these genotypes could be used in biparental mating schemes,QTL map development,and hybridization programmes to increase oil content and quality for industrial purposes.

Genetic gains with genomic versus phenotypic selection for drought and waterlogging tolerance in tropical maize(Zea mays L.)

Erratic rainfall often results in intermittent drought and/or waterlogging and limits maize(Zea mays L.)productivity in many parts of the Asian tropics.Developing climate-resilient maize germplasm possessing tolerance to these key abiotic stresses without a yield penalty under optimal growing conditions is a challenge for breeders working in stress-vulnerable agro-ecologies in the region.Breeding stress-resilient maize for rainfed stress-prone ecologies is identified as one of the priority areas for CIMMYT-Asia maize program.We applied rapid cycle genomic selection(RCGS)on two multiparent yellow synthetic populations(MYS-1 and MYS-2)to improve grain yield simultaneously under drought and waterlogging conditions using genomic-estimated breeding values(GEBVs).Also,the populations were simultaneously advanced using recurrent phenotypic selection(PS)by exposing them to managed drought and waterlogging and intermating tolerant plants from the two selection environments.Selection cycles per se(C1,C2,and C3)of the two populations developed using RCGS and PS approach and their test-cross progenies were evaluated separately in multilocation trials under managed drought,waterlogging,and optimal moisture conditions.Significant genetic gains were observed with both GS and PS,except with PS in MYS-2 under drought and with GS in MYS-1 under waterlogging.Realized genetic gains from GS were relatively higher under drought conditions(110 and 135 kg ha^(-1) year^(-1))compared to waterlogging(38 and 113 kg ha^(-1) year^(-1))in both MYS-1 and MYS-2,respectively.However,under waterlogging stress PS showed at par or better than GS as gain per year with PS was 80 and 90 kg ha^(-1),whereas with GS it was 90 and 43 kg ha^(-1) for MYS-1 and MYS-2,respectively.Our findings suggested that careful constitution of a multiparent population by involving trait donors for targeted stresses,along with elite highyielding parents from diverse genetic background,and its improvement using RCGS is an effective breeding approach to build multiple stress tolerance without compromising yield when tested under optimal conditions.

Identification of a SSR marker (TOM-144) linked to Fusarium wilt resistance in <i>Solanum lycopersicum</i>

With the discovery of molecular markers and marker assisted selection technology, the research has entered into a new era and has made it possible to develop new and more informative PCR-based markers, including SSR, and to further facilitate the use of markers in tomato breeding. The present study is a step to introduce a new SSR marker (TOM-144) which was deduced after evaluation of eight microsatellite loci amongst the twenty-one different tomato cultivars. The marker selected was inherited and segregated in mendelian fashion as demonstrated in successive generation of a cross between parent cvs. H-24 x GT-2.

Genomic analysis of a novel strain of Bacillus nealsonii, isolated from Surti buffalo rumen

Aim: Whole genome sequencing and functional annotation of Bacillus nealsonii strain AAU1, an amylolytic anaerobic spore forming isolate from ruminal contents of buffalo. Methods and Results: Morphologically, the strain was observed as slender Grampositive rods, occurring in pairs. Optimal growth was observed at 40°C (range: 30°C to 45°C) and pH 6.5 (range: 5.5 to 7.5) when cultivated in Hungate’s medium supplemented with starch. The microorganism showed extracellular constitutive amylolytic activity, proving to be capable of utilizing glucose, maltose, mannose, trehalose, dextrin and starch under anaerobic conditions. Sequence analysis revealed a GC content of 35.1 mol%. Comparison of housekeeping gene sequences for RNA polymerase subunit B (rpoB) and gyrase A (gyrA) identified sequence similarity within the Bacillus genus, confirmed by 16S rRNA gene sequence similarity which identified Bacillus nealsonii DSM 15077 as the closest publically available relative. Chemotaxonomic analysis provided conflicting results with straight-chain saturated C16: 0/C16:0 aldehyde, C16:0 DMA, C14:0 and monounsaturated 16:1w7c and 16:1w9c the major fatty acids in contrast to those reported for B. nealsonii DSM15077. Further characterization using AN-Bilog and physiological parameters provided genotypic and phenotypic support for taxonomic classification of isolate AAU1 with published Bacillus species including B. licheniformis, B. subtilis, B. circulans and B. nealsonii. Conclusion: Based on the data presented, isolate is likely to represent a new strain/subspecies, for which the identifier B. nealsonii AAU1 is proposed. Significance and Impact of Study: The strict anaerobic conditions prevailing in the bovine rumen from where AAU1 was isolated may have resulted in genetic polymorphism influencing its metabolic characteristics.

A comparative study of fermented buffalo and camel milk with anti-inflammatory, ACE-inhibitory and anti-diabetic properties and release of bio active peptides with molecular interactions: In vitro, in silico and molecular study

The aim of this study is to compare ACE inhibitory,anti-diabetic and Anti-inflammatory activity of buffalo and camel milk using Lacticaseibacillus paracasei(M11)in combination with yeast Saccharomyces cerevisiae(WBS2A)and to release peptides with antihypertensive and anti-diabetic properties.Fermented buffalo and camel milk were considered for the evaluation of Angiotensin-converting enzyme(ACE)inhibitory and anti-diabetic activities at specific time intervals(12,24,36 and 48 h)at 37℃.For the growth condition optimization,proteolytic activity was evaluated with specific inoculation rates(1.5%,2.0%,and 2.5%)and at specific incubation periods(12,24,36,and 48 h).The highest peptides were produced at 2.5%inoculation rate and at 48 h of incubation in both buffalo and camel milk at 37℃.Further,camel and buffalo milk fermented with M11+WBS2A significantly lower the overproduction of TNF-α,IL-6,IL-1βand Nitric oxide(NO),that was induced by lipopolysaccharide(LPS)stimulation in RAW 267.4 cells.For protein purification,2D gel electrophoresis and SDS-PAGE were used.Camel and buffalo milk that had not been fermented displayed protein bands between 10 and 100 kDa and 10 and 75 kDa respectively,whereas all the fermented samples showed bands between 10 and 75 kDa.On SDS-PAGE,in permeates,there were no apparent protein bands.When fermented buffalo and camel milk were electrophoresed in 2D gel,a total of 26 and 25 protein spots were found.In 2D gel electrophoresis,protein spots had sizes between 10 and 37 kDa.In RP-HPLC,water-soluble extracts(WSEs)of ultra-filtered fractions were used to distinguish different fractions of a peptide.Peakview software was used to characterize the LC/MS results.Fourier transform-infrared spectroscopy(FTIR)was used to evaluate various functional groups of fermented milks.In molecular docking study,the peptide sequences identified from the fermented buffalo milk(FBM)and camel milk(FCM)were examined for their binding affinity against the binding sites of hPAM and hMGA.

Factors influencing farmers’adoption of solar water-pumping systems in Gujarat

Agriculture has played an important role in the growth of the Indian economy.Water and electricity are essential inputs for agriculture today.India produces the third-largest amount of electricity in the world,behind China and the USA.As an incentive to increase production,most states provide free electricity to farmers.Free electricity has decreased groundwater levels and increased electricity consumption.The objective of this study was to find out the factors that influenced farmers to adopt a solar water-pumping system in Gujarat.The samples were randomly selected.This study examined the entire state of Gujarat.One hundred and fifty farmers who adopted solar water-pumping systems were interviewed,including 50 banana farmers,50 cotton farmers and 50 groundnut farmers.Primary data were collected through an interview.The adoption of solar water-pumping systems by farmers was identified using factor analysis.SPSS software was used to analyse the data collected.In this study,only two factors contributed to the variance of 59.469%.The adoption of solar water pumps by farmers was influenced by government policy and economic benefits.An economic benefit variance of 33.20%was found,while a government policy variance was 26.27%.A solar water-pumping system has low initial costs,flexible credit policies motivate adoption,solar water-pumping systems can save electricity,and maintenance and repair costs are affordable.Government policy that includes solar water-pumping systems is motivated by awareness,subsidies for adoption and a fair price to sell energy surpluses.Farmers are recommended to adopt a solar water-pumping system in conjunction with a drip irrigation system to receive additional income.The successful scheme will be recommended to other states in India for implementation.

Scenario of solar energy and policies in India

Solar power is energy from the Sun that is converted into thermal or electrical energy.China,the USA,and India have the world’s richest solar energy sources.India receives solar radiation with an average intensity of 200 MW/km^(2) and 250–300 sunny days a year.The solar radiation in India varies from 4 to 7 kWh/m^(2)/day,with 2300–3200 sunshine hours yearly,depending on the region.Badhla Solar Park,Jodhpur,India,is the world’s largest solar plant,with 2.25 GW of installed capacity.Solar energy is the best alternative elec-tricity source used explicitly in the agricultural sector.In the farming sector,the consumption of solar energy has been increasing.The major issues were the installation costs,low availability of technicians,and poor quality of panels.The focus on skill development equips the workforce to drive innovation,efficiency,and sustained growth in the renewable-energy field.With the support of inter-national collaboration and a skilled workforce,India’s progress in solar energy development is a boon for the industry.

Effect of Storage Temperature on Vitamin C, Total Phenolics,UPLC Phenolic Acid Profile and Antioxidant Capacity of Eleven Potato(Solanum tuberosum) Varieties

Storage of potato tubers at low temperature affects their metabolism and may alter their phytochemical properties. There is a need to elucidate the changes in antioxidant compounds, activity and enzymes during storage of tubers. Eleven Indian potato varieties were evaluated for antioxidant parameters, after 0, 30, 60 and 90 days of storage at room temperature, 15 °C and 4 °C. Total phenolics(0.0786–0.1546 mg gallic acid equivalents·g^(-1)FW) and vitamin C content(0.0828–0.2416 mg·g^(-1)FW) varied among the varieties and were different with storage temperature; their levels fluctuated during storage but remained above the initial level until the last day of observation. Phenolic acid profiling by UPLC identified 12 compounds among which the most abundant was chlorogenic acid followed by gallic acid, sinapic acid and ellagic acid. Except para-coumaric acid which decreased at 4 °C, all the phenolic acids increased with storage. Caffeic acid, chlorogenic acid, protocatechuic acid and gallic acid mostly correlated with total phenolic content(r = 0.456, 0.482, 0.588 and 0.620, respectively). Antioxidant activity against both DPPH and ABTS radicals increased during the initial days of storage and then dropped to a level comparable or lower than the original value,irrespective of the storage temperature. Correlation study revealed that chlorogenic acid, gallic acid and ferulic acid mostly contributed to antioxidant activity. Activity of both antioxidant enzymes, superoxide dismutase and ascorbate peroxidase, increased initially but then decreased to values lower than the initial level and were not influenced by storage temperature. Correlation with antioxidant activity indicated that the enhancement of reactive oxygen scavenging species in cold stored tubers could result mainly from ascorbate peroxidase activity. Our results demonstrate that storage temperature adversely influences the metabolism and the content of antioxidant compounds in potato tubers, with subsequent increase on their antioxidant capacity.

Air flow characteristics of an air-assisted sprayer through horizontal crop canopy

Artificially induced air currents or air-assistance to droplet spectrum produced by hydraulic nozzles not only facilitate in transporting and depositing the droplets in different parts of canopy but also reduce the application rate of chemicals.The air streams increase the velocity of smaller droplets so that extra momentum would increase impaction and improve penetration into the crop as well as mitigating the influence of wind on drift.It is necessary to quantify the airflow characteristics.But,control of climatic and other conditions in the field is very difficult.Thus,airflow characteristics study was done under controlled conditions on a horizontal simulated crop canopy.Based on this study,an airflow distribution model was developed and airflow characteristics for vegetable crops,namely,eggplant,chilli and bittergourd were predicted.The differences between predicted and actual field study values were not statistically significant.Kinetic energy of air stream dissipated with its movement from top to bottom of the canopy.The rate of kinetic energy dissipation was higher in denser canopies.Higher air velocity 15 m/s was the best as it produced maximum turbulence throughout the canopy.

Optimization of enzymatic hydrolysis of pigeon pea for cooking quality of dhal

Cooking quality of dehusked splits is influenced by the dehulling method,in particular,by the pre-milling treatments.The effects of four enzymatic hydrolysis parameters,i.e.,enzyme concentration(20-60 mg/100 g dry matter),incubation time(3-15 h),incubation temperature(40-60℃)and tempering water pH(4.0-6.0)on cooking time of pigeon pea dhal were optimized using response surface methodology.Three kinds of enzymes,i.e.,xylanase,pectinase,and cellulas were used in combination for enzymatic pre-treatment.A quadratic model satisfactorily described the dehulling efficiency with high value for the coefficient of determination R^(2)(0.9062).It predicted a minimum cooking time of 21.91 min at enzyme concentration of 37.8 mg/100 g dry matter,incubation time 8.69 min,incubation temperature 48.5℃ and pH 5.49 of tempering water.Cooking time at optimum condition was observed to be 21.50 min and the predicted values of cooking time showed 2.19% deviation from the experimental values.Results of the study revealed that cooking time of enzyme treated dhal could be decreased by 19.77% compared to the oil treated dhal.

Tissue-specific Temporal Exome Capture Revealed Muscle-specific Genes and SNPs in Indian Buffalo(Bubalus bubalis)

Whole genome sequencing of buffalo is yet to be completed, and in the near future it may not be possible to identify an exome （coding region of genome） through bioinformatics for designing probes to capture it. In the present study, we employed in solution hybridization to sequence tissue specific temporal exomes （TST exome） in buffalo. We utilized cDNA prepared from buffalo muscle tissue as a probe to capture TST exomes from the buffalo genome. This resulted in a prominent reduction of repeat sequences （up to 40%） and an enrichment of coding sequences （up to 60%）. Enriched targets were sequenced on a 454 pyro-sequencing platform, generating 101,244 reads contain- ing 24,127,779 high quality bases. The data revealed 40,100 variations, of which 403 were indels and 39,218 SNPs containing 195 nonsyn- onymous candidate SNPs in protein-coding regions. The study has indicated that 80% of the total genes identified from capture data were expressed in muscle tissue. The present study is the first of its kind to sequence TST exomes captured by use of cDNA molecules for SNPs found in the coding region without any prior sequence information of targeted molecules.

Residue determination ofβ-cyfluthrin and imidacloprid as mix formulation in/on chickpea(Cicer arietinum)pods and soil and its risk assessment

OBJECTIVES:A field experiment was conducted at Agricultural Research Station for irrigated crops at Thasara,Anand Agricultural University,Anand,to study the dissipation and risk assessment ofβ-cyfluthrin and imidacloprid as combination product in/on chickpea.MATERIALS AND METHODS:Solomon 300 OD comprising 9 per centβ-cyfluthrin and 21 per cent imidacloprid was foliar sprayed at the doses of 18+42g a.i.ha−1(standard dose)and 36+84g a.i.ha−1(double dose).Totally three sprays were made at an interval of 1 week starting from fruiting stage.RESULTS AND DISCUSSIONS:The residues ofβ-cyfluthrin estimated using gas chromatograph with electron capture detector(GC-ECD)showed an initial deposit of 0.16 and 0.27μg g−1 in standard and double doses,respectively.The residues for corresponding doses persisted till 7th and 10th day after application and reached below determination level of 0.01μg g−1 on the 10th and 15th day,respectively.Imidacloprid was estimated using high performance liquid chromatography(HPLC)and showed initial deposits of 1.22 and 2.7μg g−1 in standard and double doses,respectively.Its levels reached below the determination level(0.01μg g−1)on the 15th day in green pods.Statistical analysis of dissipation kinetics showed thatβ-cyfluthrin followed zero-order kinetics in standard dose with half-life of 7.27 days.In double dose,the dissipation kinetics followed first-order kinetics with half-life of 9 days.Imidacloprid followed first-order kinetics in both the doses with half-life of 6.7 and 7.7 days.CONCLUSIONS:A pre-harvest interval(PHI)of 15 days is suggested.Theoretical risk assessment calculated as hazard quotient was less than 1,rendering the use of combination product as safe-provided proper PHI is followed.

Bioactivities and ACE-inhibitory peptides releasing potential of lactic acid bacteria in fermented soy milk

This study was designed to evaluate the bioactivities such asβ-glucosidase activity,α-galactosidase activity,and the growth behavior of the Lactobacillus cultures in soy milk medium.Ten Lactobacillus cultures were considered in this study.L.fermentum(M2)and L.casei(NK9)were selected due to their betterα-galactosidase,β-glucosidase activity and growth behavior in soy milk medium during fermentation.Further,soy milk fermented with M2 showed higher proteolytic activity(0.67 OD)and ACE-inhibitory(48.44%)than NK9(proteolytic activity:0.48 OD and ACE-inhibitory activity:41.33%).Bioactive peptides produced during the fermentation of soy milk using the selected Lactobacillus cultures were also identified with potent ACE-inhibitory activity by MALDI-TOF spectrometry,and the identified ACE inhibitory peptide sequences from fermented soy milk were characterized using Biopep database.

Exploring the potential of Lactobacillus and Saccharomyces for biofunctionalities and the release of bioactive peptides from whey protein fermentate

Whey protein concentrate-80(WPC-80)fermented with L.fermentum(KGL4)(37℃)and S.cerevisiae(WBS2A)(25℃)was tested for ACE-inhibitory and antioxidant activities over different periods(12,24,36 and 48 h).Proteolytic activity(OPA method)was used to optimize the growth conditions(inoculation rate,i.e.at 1.5%,2.0%,and 2.5%and incubation time,i.e.12,24,36,and 48 h)for peptide production.Results indicated that the highest amount of peptides was obtained at 7.24 mg/mL for KGL4(37℃,48 h)and 8.59 mg/mL for WBS2A(25℃,48 h).The whey protein fermentate inhibited the LPS-induced NO production,while enhanced production concentrations of TNF-α,IL-6,and IL-1β.Subsequently,SDS-PAGE,as well as Two-Dimensional(2D)gel electrophoresis methods,were applied for protein purification using water-soluble extracts(WSEs)of WPC-80 fermented by a combination of L.fermentum and S.cerevisiae.On SDS-PAGE,protein bands were observed in the range of 10-55 kDa,whereas on the 2D page,protein spots were in the range of 10-70 kDa.All the 2D spots were analyzed using RPLC/MS.WSEs of 3 kDa and 10 kDa permeates were used in RP-HPLC to identify distinct peptide fractions.The data from LC/MS was also characterized by utilizing ProteinPilot software.Further,different functional groups were also analyzed using FTIR investigation.The research aims to isolate and characterize novel ACE-inhibitory and antioxidative peptides from fermented WPC-80 produced by Lactobacillus fermentum and S.cerevisiae.

Antioxidative,antimicrobial and anti-inflammatory activities and release of ultra-filtered antioxidative and antimicrobial peptides during fermentation of sheep milk:In-vitro,in-silico and molecular interaction studies

The antioxidative,antimicrobial,and anti-inflammatory properties of fermented sheep milk with L.fermentum (KGL4),as well as the generation of antioxidative and antimicrobial peptides,are evaluated in the study.Antioxidative and antibacterial activities in sheep milk fermented with KGL4 increased with incubation hours,along with varied antioxidative properties (ABTS assay:35.12%,hydroxyl free radical scavenging assay:29.12%,superoxide free radical scavenging activity:36.38%).Antimicrobial activity of fermented sheep milk against E.faecalis (19 mm),S.typhimurium (15.67 mm),B.cereus (14 mm),and E.coli (13 mm) was also observed.Furthermore,after 48 h,the KGL4 showed maximum proteolysis (10.40 mg/ml) at 2.5% rate of addition sheep milk.The antioxidative and antimicrobial activities of fermented sheep milk fractions (3 kDa and 10 kDa permeates and retentates) were also investigated.The highest ABTS activity (26.90%) was found in 10 kDa permeate,whereas the 3 kDa retentate had higher hydroxyl free radical scavenging activity (69.20%) and 3 kDa permeate showed maximum superoxide free radical scavenging activity (32.85%).The 10 kDa retentate had shown maximum antimicrobial activity against S.typhimurium (13.67 mm) and E.faecalis (17.00 mm).Proteins spotted on 2D gel electrophoresis of KGL4 were varied from 10 to 70 kDa.RP-LC/MS was used to identify 5 novel peptide sequences from 2-D gel spots.Searches in the BIOPEP database confirmed the antioxidative and antimicrobial effects of the novel fermented sheep milk peptides.Fermented sheep milk with KGL4 (SMKGL4) significantly reduced excessive TNF-α,IL-6,& IL-1β production in LPS-activated RAW 264.7 cells.