Sajeshkumar N.K. – författare

Scientific Study from the year 2016 in the subject Agrarian Studies, Mar Augusthinose College (Mar Augusthinose College), course: Biotechnology, language: English, abstract: Urban and peri-urban agriculture (UPA) provide a significant role in ensuring urban food security, income generation and livelihood strategies and supports Millennium Development Goals (MDGs). Quantitative data about the phytoavailability and food chain transfer of heavy metals in Mumbai Metropolitan Region (MMR) is scarce. This study was conducted to characterize the elemental and heavy metal transfer among major UPA production systems (farms and railway gardens), in MMR eliciting the soil to root translocation as well as its localization in produce. It comprises a detailed two year onsite examination of three farms (F1-3) and three railway gardens (RG1-3) across MMR. Potential risk assessments were conducted by metal transfer factor (MTF), metal translocation (TF), daily intake of metals (DIM), health risk index (HRI), average daily dose (ADD) and target hazard quotient (THQ) as well as total metal and element content in comparison with different safety standards. Copper concentration in soils ranged from 29.7 - 545.1 mg kg-1, with highest and lowest concentrations observed at RG3 and RG2 respectively. The shoots of white radish accumulated Sr concentrations up to 424.1 mg kg-1 at RG2. Strontium had a TF up to 32.25 in comparison with Co, Cu, Zn, Ni and Cr with a maximum of 5.93, 5.32, 3.41, 1.71 and 1.47 respectively. Average daily dose of Zn was between 1.3 10-1 and 3.6 10-1 mg kg-1 d-1 while Ni had a daily dose of between 4.6 10-2 and 7.4 10-2 mg kg-1 d-1. The estimated values of ADD were below the world standard levels except for Zn and Ni, there is a relative absence of health risk imposed by the ingestion of these vegetables produced in UPA systems in MMR.

Scientific Study from the year 2016 in the subject Agrarian Studies, , language: English, abstract: Jackfruit (Artocarpus heterophyllus) is commonly grown in home gardens of tropical and sub-tropical countries. The fruit contains high levels of carbohydrates, protein, starch, calcium and vitamins. The Artocarpus heterophyllus is believed indigenous to the Western Ghats of India. It is adapted only to the humid tropical and near tropical climate. The tree flourishes in rich deep soil of medium or open texture, sometimes on deep gravelly or laterite soil. In India they say that the tree grows tall and thin on sand, short thick on sandy land. Here the physical and chemical properties of soil from surrounding plots of different varieties of Arthocarpus heterophyllus have been analysed by using different technical methods and evaluating the effects of soil property on jackfruit nutritional quality. It was observed that there was a considerable difference in case of some minerals likes phosphorous, potassium, magnesium, zinc and manganese. Change in the soil pH also noticed. The pH also plays a major role in the sweetness of the jackfruit. The optimum pH of the amylase inside the jackfruit was found to be 6.2 to 6.8. The result shows that the soils that have higher pH have more sweet flakes. Proximate nutritional composition of the flake reveals the presence of protein, fat and fibre in a greater amount in some flake and these flakes were sweeter than the others. There was a much difference in the sugar level also. Increased protein, fat and fibre content found in flake of the trees that were growing in soil that have higher concentration of minerals. Further studies such as molecular studies, enzymatic studies should conduct to identify the whole variation among jackfruit in Kerala. Encouragements should be done to the marketing as well as value added food products from this underutilized fruit tree.

Scientific Study from the year 2016 in the subject Biology - Botany, Mar Augusthinose College, language: English, abstract: Diabetes is a complex metabolic disorder resulting from either insulin insufficiency or insulin dysfunction. Diabetes mellitus affects most of the people in both developed and developing countries. The treatment of diabetes with synthetic drugs is costly and chances of side effects are high. Phytomedicine has been used since ancient times in various parts of the world where access to modern medicine is limited. Medicinal plants and phytoconstituents play an important role in the management of diabetes mellitus especially in developing countries where resources are meagre. Phytochemicals identified from medicinal plants present an exciting opportunity for the development of new types of therapeutics for diabetes mellitus. Most prevalent among phytochemical groups are the alkaloids, glycosides, polysaccharides, and phenolics such as flavonoids, terpenoids and steroids. This study aims to provide a comparative information about various plants used for antidiabetes treatment in a trial community (Mannans) and their constituents, which have been shown to display potent hypoglycemic activity. The current study was focused on four plants named Tinospora cordifolia, Ensete superbum, Coccinia grandis wild and Apama siliquosa Lam. which is locally known as Amruthu, Kalluvazha, Kattukoval, and Alpam respectively. The plant parts such as seed, leaves, stem and bark were used by the local people. The plant materials prepared as decoction, infusion, aqueous extracts in milk or honey were used for the treatment of diabetes. The plants that are being used by the local people of the study area have been isolated from plants for the treatment of diabetes. The efficacy of these ethnomedicinal plants needs to be subjected to pharmacological validation. it was found that Coccinia grandis was more efficient than the other plants. Some antidiabetic plants may exert their action by stimulating the function or number of beta-cells and thus increasing insulin release. The study highlighted the central role of traditional herbal medicine for the treatment of diabetes in tribal community. Ethnobotanical survey is most useful for scientists, research scholars and scientific companies for further studies on isolation and identification of active compounds that can be formulated into antidiabetic drugs.

Scientific Study from the year 2017 in the subject Biology - Genetics / Gene Technology, Mar Augusthinose College, language: English, abstract: Artocarpus heterophyllus belong to the Moraceae family and are abundant in Western Ghats. The fruit provide two MJ per kg/wet weight of ripe perianth and contain high levels of carbohydrates, protein, starch, calcium and vitamins. Jackfruit has diverse medicinal uses especially for anti-oxidant, anti-inflammatory, antimicrobial, anti-cancer and anti-fungal activity. MatK (maturase K) genes are fast evolving, highly variant regions of plant chloroplast DNA that can serve as potential biomarkers for DNA coding and also in generating primers for plants with identification of unique motif regions. Advances in the genetic markers such as RFLP and PCR based methods are more reliable for identification of genetic diversity than morphological markers, although each technique has advantages and limitations. The objective of this research work was to estimate the level of genetic diversity and to assess genetic relationships among six varieties of jackfruit using matK gene based on PCR technique and RFLP markers. The partial sequence of the matK gene of six different Artocarpus varities was used in the analysis. The size of amplified products was approximately 700 bp. After sequencing and sequence editing, sequence information on a 674 bp region was finally obtained for analysis. The alignment of sequences revealed two haplotypes out of 674 sites. The nucleotide frequencies are 30.00% (A), 37.69% (T/U), 17.93% (C), and 14.39% (G). Being one of the underutilized fruits in India, Artocarpus heterophyllus Lam. has promising leads to further scientific researches and livelihood strategies. The study of matK gene using PCR and RFLP seems to a promising tool in establishing genetic diversity among jackfruit varities. The tree, indigenous to the Western Ghats, is an important source of nutritious food during summer season. Encouragements should be made to the marketing as well as value added food products from this underutilized fruit tree.

Scientific Study from the year 2017 in the subject Agrarian Studies, Mar Augusthinose College, language: English, abstract: The Cucurbitaceae family is the one of the economically important groups of plants in the tropics and subtropics. Molecular phylogenetic analysis was advanced after the introduction of molecular markers which give much precise results in analysis. Our current study based on the amplification of RuBisco enzyme using rbcL primer and subsequent validation using BLAST, FASTA and CLUSTAL-W in pumpkin and winter melon. The isolated and purified DNA samples were PCR amplified using rbcL primer and later sequenced using ABI Prism 377 DNA sequencer. Multiple sequence alignment algorithms and distance matrix were constructed using rbcL sequences in FASTA format were retrieved from GenBank. Phylogenetic tree was created using the distance based neighbour joining (NJ) and clustering algorithms method. The agarose gel electrophoresis were used to separate the isolated as well as PCR amplified DNA samples. The sequences obtained after sequencing were subjected to BLAST similarity search and multiple sequence alignment using CLUSTAL-W. The construction of divergence matrix and phylogenetic dendrograms revealed two main groups, one consisting of MC and other Momordica species while the other group consisted of three subgroups of CM, CG and BH having members of Cucurbita, Coccinia and Momordica respectively. The reliability of molecular phylogenesis can be affected by myriad, long branch attraction, saturation and taxon sampling problems. So the selection of the models plays a key step in the success analysis. In general, the present study unambiguously confirmed the existence of Coccinia grandis as a distinct species and morphologic separation between Coccinia grandis and other selected members of Cucurbitaceae as separate species was evident. Among the species included in the present study Coccinia grandis undergone maximum divergence during the process of evolution. More detailed work on the evolution within the Cucurbitaceae needed for crop improvement.

Scientific Study from the year 2016 in the subject Biology - Genetics / Gene Technology, Mar Augusthinose College, language: English, abstract: Jackfruit (Artocarpus heterophyllus) is commonly grown in home gardens of tropical and sub-tropical countries. The fruit contains high levels of carbohydrates, protein, starch, calcium and vitamins. Jack fruit has diverse medicinal uses, especially anti-oxidant, anti-inflammatory, anti-microbial, anti-cancer and anti-fungal activity. Jackfruit is considered to be an underutilized fruit where most of the fruits get wasted due to ignorance, lack of post harvest technology and gaps in supply chain systems. Jackfruit contains more protein, calcium, iron, vitamins and other essential nutrients as compared to the common fruits. To find out the morphological variation among jack fruits among Kerala we conducted an elaborative survey and found out the parameters such as tree characteristics, leaf characterises or fruit characteristics of selected samples. Both qualitative and quantitative data are measured using field visits and standard methods and morphological indicators; after analysis of data there is a morphological variation among the jack fruit samples identified. Advances in the genetic markers such as RFLP and PCR based methods are more reliable for identification of genetic diversity than morphological markers although each technique has advantages and limitations. The objective of this research work was to estimate the level of genetic diversity and to assess genetic relationships among six varieties of jackfruit using the rbcL gene based on PCR technique and RFLP markers. The partial sequence of rbcL gene of six different Artocarpus varieties was used in the analysis. The size of amplified products was approximately 700 bp. After sequencing and sequence editing, sequence information on a 651 bp region was finally obtained for analysis. The alignment of sequences revealed two haplotypes out of 651 sites. The nucleotide frequencies are 27.96% (A), 29.47% (T/U), 19.69% (C), and 22.89% (G). Being one of the underutilized fruits in India, Artocarpus heterophyllus Lam has promising leads to further scientific research and livelihood strategies. The study of rbcL gene using PCR and RFLP seems to be a promising tool in establishing genetic diversity among jackfruit varieties.

Scientific Study from the year 2017 in the subject Agrarian Studies, Mar Augusthinose College, language: English, abstract: The Cucurbitaceae family is the one of the economically important group of plants in the tropics and subtropics. Molecular phylogenetic analysis were advanced after the introduction of molecular markers which give much precise results in analysis. Our current study based on the amplification of RuBisco enzyme using rbcL primer and subsequent validation using BLAST, FASTA and CLUSTAL-W in pumpkin and winter melon. The isolated and purified DNA samples were PCR amplified using rbcL primer and later sequenced using ABI Prism 377 DNA sequencer. Multiple sequence alignment algorithms and distance matrix were constructed using rbcL sequences in FASTA format were retrieved from GenBank. Phylogenetic tree was created using the distance based neighbour joining (NJ) and clustering algorithms method. Once divergences between all pairs of samples were determined, statistical cluster analysis and dendrogams examines the similarity among halotypes. Bootstrapping and jackknifing further increase the reliability estimates for the position of haplotypes within the evolutionary tree.

Scientific Study from the year 2016 in the subject Agrarian Studies, grade: 1.5, Mar Augusthinose College, language: English, abstract: Glyphosate (N-phosphonomethylglycine) is a herbicide that is used worldwidely. Its common trade name is Roundup. Its non-targeted species action makes it most popular herbicide. It was developed by Monsanto company. The primary target for glyphosate is the enzyme EPSPS (5-enolpyruvoylshikimate 3-phosphate synthase). When glyphosate binds to EPSPS it forms a very stable complex that essentially permanently disables the enzyme and hence affect the metabolic activity of the plant and results in its death. Finding Glyphosate degrading microorganisms from soil is an interesting topic since glyphosate is non- targeted in its toxicity. Microorganisms were isolated from soil samples, which were then identified by molecular method. Isolation of DNA, its amplification using 16s rRNA gene and its sequencing are the major steps involved. Bioinformatics tool helps to identify the microorganisms. Two microorganisms identified are Pseudomonas sp. and Achromobacter sp. In the phylogenetic analysis also the two organisms are grouped as separate clads. In these, strain 1 showed highest growth in the Glyphosate containing medium than strain 2. These results show that the bacterial strain may possess potential to be used in bioremediation of glyphosate-contaminated environments.

Scientific Study from the year 2017 in the subject Biology - Micro- and Molecular Biology, grade: 1.5, Mar Augusthinose College, language: English, abstract: The dyes are complex chemical compounds that imparts colour to substances. On the urge of urbanisation, the use of synthetic dyes is increasing largely and its untreated effluent release causes serious environmental pollution affecting water bodies by disturbing the aquatic ecosystem, soil, plants, animals, humans. The recalcitrant nature of these dyes limits its treatment using conventional methods wherein biological methods using microorganisms are reported to completely mineralize the dyes lowering the release of degradation products. The present work focuses on the biodegradation of synthetic dyes by isolating bacteria from a dyeing unit. Soil samples from the dye contaminated soils was collected, its degradation potentiality was observed using three major dye of studies congo red, brilliant green and methylene blue within 24 hr incubation. Maximum decoloarized dye (congo red and brilliant green) were chosen, serially diluted to 10-5 and plated to obtain two distinct colonies from decolorized congo red (CR1, CR2) and three distinct colonies from decolorized brilliant green (BG1, BG2, BG3). These isolates were biochemically characterized. Molecular characterization was performed by isolating DNA from five isolates and amplified it using PCR, with the 16s rRNA gene primer. The PCR amplification product having approximately 1500bp were sequenced, edited and searched using BLAST against the known sequences within NCBI databases. The isolates were identified to be CR1 as Pseudomonas (88% identity), CR2 as Aeromonas (89% identical) and BG1/BG2 were confirmed to belong to same genus as Bacillus (99% identical). The phylogenetic tree showed a clear divergence between isolated species. Furthermore, the dye tolerance of isolates were observed by optimization analysis to be as CR1 and CR2 tolerated up to 4% of congo red dye and among BG1, BG2 and BG3; BG1 tolerated up to 0.50% of brilliant green dye. Thus, CR1 and CR2 were observed to be potent azo dye decolourizers. Recombinant DNA Technology can be applied in this field that can make the above future application more reliable.

Scientific Study from the year 2016 in the subject Chemistry - Bio-chemistry, grade: 1.5, Mar Augusthinose College, language: English, abstract: Amylase is an enzyme which catalyzes the hydrolysis of a (1, 4)-glycosidic linkages in amylose (a linear form of starch), amylopectin (a branched form of starch) and glycogen into simpler carbohydrate molecules such as oligosaccharides or disaccharides. Alpha-amylase is the major form of amylase found in human, most prominently in pancreatic juice and saliva. The salivary amylase is an amylolytic enzyme, which can acts on cooked or boiled starch and converts it in to maltose. So it became interesting to study the behaviour of salivary amylase, when it is secreted as result of different stimuli. And thus began to study the effect of five different stimulatory temperatures, and also the effect of four tastes on the behaviour of salivary amylase. For the study of stimulatory effect of temperature on salivary amylase, five different temperatures are selected (4, 27, 37, 55 and 75 C). And likewise four tastes also selected (sweet, sour, salt and bitter). The DNS method was done in the both tests to obtain the absorbance at 520 nm. The samples were collected from three people, of same age. The saliva was collected at same time, after one and a half hour of their breakfast in order to maintain a controlled condition for this study. In each cases the incubation temperature also kept as variable (4, 27, 37, 55 and 75 C). This study was also aimed to determine the behaviour of salivary amylase with reference to the kinetic parameters like Km and Vmax of salivary alpha amylase by incubating the enzyme (stimulated by different stimulatory conditions of temperature and taste) with varying concentration of substrate. The study revealed the consistency in kinetic parameters like Km and Vmax of salivary alpha amylase secreted in response to various stimuli.

Scientific Study from the year 2016 in the subject Agrarian Studies, grade: 1.5, Mar Augusthinose College, language: English, abstract: Jackfruit (Artocarpus heterophyllus) is commonly grown in home gardens of tropical and sub-tropical countries. The fruit contains high levels of carbohydrates, protein, starch, calcium and vitamins. Jack fruit has diverse medicinal uses especially as anti-oxidant, anti-inflammatory, antimicrobial, anti-cancer and anti-fungal agent. Jackfruit is considered to be an underutilized fruit where most of the fruits get wasted due to ignorance, lack of post-harvest technology and gaps in supply chain systems. Jackfruit contains more protein, calcium, iron, vitamins and other essential nutrients, compared to common fruits. To find out the morphological variations among jackfruits in Kerala we conducted an elaborate survey and found out varieties using parameters such as tree characteristics, leaf characterises, fruit characteristics of selected samples as measure. Both qualitative and quantitative data were measured using field visit and standard methods and morphological indicators. After analysis of data morphological variations among the jack fruit samples were identified. This includes the morphological difference between fruit stalk leaf and branch leaves. In the fruits themselves, colour, shape and size are also found to vary among the samples. These variations may have some relationship to climatic factors such as rainfall, pH and soil properties. Morphological variations affect the selection of the varieties. Encouragement should be given to the marketing as well as creation of value added products from this underutilized fruit tree.

Scientific Study from the year 2016 in the subject Agrarian Studies, grade: 1.5, Mar Augusthinose College, language: English, abstract: This study aims at the attributes of the Annona reticulata and its medical and biological value. Annona reticulata belongs to the family Annonaceae, commonly known as honey apple. Qualitative phytochemical analysis of chloroform and water extracts of Annona reticulata fruit, leaf and stem bark was conducted in order to detect the presence of various secondary metabolites using standard procedures. The results of phytochemical screening indicated the presence of secondary metabolites such as tannins, betacyanins, carbohydrates, alkaloids, terpenoids, phenols, quinines, saponins, cardiac glycosides etc. Also the comparative antimicrobial activity of chloroform and water extracts of fruit, leaf and stem bark of Annona reticulata was evaluated against four bacterial species namely, Escherichia coli, Pseudomonas aeruginosa, Serratia marcescens and Micrococcus luteus and two fungal species namely Candida albicans and Rhizopus. Agar well diffusion method and disc diffusion method were selected to check the antimicrobial activities of the extracts. The study revealed that the chloroform extracts of leaf, stem bark and fruit of Annona reticulata has activity against the bacterial strains and fungal strains. Whereas, the water extracts of leaf, fruit and stem bark of Annona reticulata has more activity towards the fungal species. The findings of this study have identified that Annona reticulata extracts acts as a promising source of antimicrobial agent which could be useful in the modern medicine.

Scientific Study from the year 2017 in the subject Chemistry - Bio-chemistry, grade: 1.5, Mar Augusthinose College, course: Biotechnology, language: English, abstract: Diabetes mellitus is a clinical metabolic syndrome, which has been resulted from the malfunction or insufficiency of insulin production. Diabetes is the major threat to the people in developing and developed country. Prolonged diabetic condition can affect organs and synthetic drug if consumed by the patient can cause side-effects and impaired immune system. The synthetic medicine used for the treatment of diabetes is usually costlier, create side-effects and do not ensure complete recovery from the disorder but on the other hand, medicinal plants create a great impact in the human immune system. The medicinal plant produces a variety of chemical substances which shows significant therapeutic properties. This study highlights the potential anti-diabetic significance and phytochemical activity of different varieties Piper nigrum (Black pepper) and different strategies used for the assessment of these activities. The piperaceae family consist of 12 genera and more than 1400 species and is one of the largest families and widely distributed across the tropical and sub-tropical regions and over the high ranges of Andes and Sub Himalayas. Piper species naturally have multi-dimensional affect on various parts of the body often mentioned as antipyretic, diuretic, aphrodisiac, immune- stimulant and antioxidant, hepatoprotective, digestive, antiseptic and anti-diabetic. Phytochemical analysis of Methanol and Petroleum -ether extracts was carried out and different plant parts were collected from different districts across Kerala. Phytochemical which is identified from Piper nigrum present an exciting opportunity for the development of new type of therapeutics for diabetes mellitus. Some anti-diabetic plants exert their action by stimulating the function or number of beta cells and thus increasing insulin production with least side effects. Further investigations are required and more attention should be drawn to explore the biological activity of hundreds of traditionally used plants. Both in-vivo and in-vitro assessment is necessary for the assessment of anti-diabetic properties from the natural resources.

Scientific Study from the year 2017 in the subject Agrarian Studies, grade: 1.5, Mar Augusthinose College, course: Biotechnology, language: English, abstract: Black pepper belongs to piperaceae family and is known as "e;king of species"e;. This piperaceae family contain approximately 2,000 species. Phosphorus is one of the most important micronutrients and they are essential for the biological growth, development of plants and it is the most essential nutrient for plants. Phosphate deficiency is wide spread and phosphate fertilizers universally required in the form of inorganic P fertilizers, only a small portion is utilized by plants and the remaining are in insoluble form and they are solubilised by the microbes present in the soil. Soil, they are rich in micro and macronutrients and sixteen elements or nutrients are essential for plant growth and reproduction. Several soil bacteria, particularly belonging to the genera Pseudomonas and Bacillus posses ability to bring insoluble soil phosphate into soluble forms by secreting acids like formic, lactic and acetic. The rhizosphere soil sample were serially diluted up to 10-4 using sterile distilled water and plated on Pikovskaya s agar medium by pour plate method. The P solubilising isolates was evaluated on agar plates of Pikovaskya growth medium by solubilising the tricalcium phosphate of the medium. After incubation the phosphate solubilising microorganisms were selected based on different colonies. Out of so many bacterial isolates, 8 isolates were selected for the further study to perform qualitative and quantitative analysis. Phosphate solubilising microorganisms are possible to use as bio fertilizer for all crops. Among the isolates some of are almost identical in biochemical test but they have different morphology characters.

Scientific Study from the year 2016 in the subject Biology - Micro- and Molecular Biology, grade: 1.5, Mar Augusthinose College, language: English, abstract: In this study an attempt was made to evaluate the colour degradation capabilities by collecting the contaminated soil sample from Kalady area and serial dilution was done upto 10-6. From the dilution 10-5 was taken and spread plated on Nutrient agar. From the above plate, isolated colonies was obtained which was found to be Bacillus sp and Pseudomonas sp respectively by morphological, microscopical and biochemical method. The isolated colonies was taken for degradation studies with 1% dye and 1% inoculum in Nutrient broth and OD values and colour change was noted. It was found to be Bacillus sp has more degrading capacity in yellow colour than Pseudmonas sp. The optimization studies was done with Bacillus sp having different concentration of colour (2, 4, 6) with varying pH (4,6,8) and temperature (37(deg)C, 40(deg)C and room temperature). The result was found to be having the concentration of colour with 4% having pH 4 and temperature 37(deg)C.

Scientific Study from the year 2017 in the subject Chemistry - Bio-chemistry, grade: 1.5, Mar Augusthinose College, language: English, abstract: As the prevalence of obesity and hypercholesterolemia are very common in our society, plants with cholesterol lowering action has great value in modern therapeutics. The phytochemicals present in the extracts of Averrhoa bilimbi were analyzed and its effect on lowering cholesterol in various fatty food materials was evaluated in vitro. Various phytochemical compounds like tannins, saponins, alkaloids, emodins, proteins, carbohydrate, terpenoids, glycosides, flavonoids, coumarins and phenols were found in the fruit extracts of the plant. The level of cholesterol was evaluated by Zak's method in five different fatty food materials. After the treatment with extract four of them showed significant reduction in the cholesterol level day by day and no change in the cholesterol level was observed in one sample.

Scientific Study from the year 2017 in the subject Chemistry - Bio-chemistry, grade: 1.5, Mar Augusthinose College, course: Biotechnology, language: English, abstract: Aqueous extract of the fruit pulp of Tamarind (Tamarindus indica) were evaluated for cholesterol lowering effect, in vitro, against various fatty food materials. People consume food items made by chicken, beef, mutton, egg and fish and it contains large amount of fat. This study aims to analyze the effect of Tamarindus indica in reducing the cholesterol level in this fat compound using water extract of the pulp. For this fatty food samples like egg yolk, pork fat, chicken fat, ghee and cod liver oil were treated with the extract and cholesterol level was estimated by Zak s method for a period of time. Phytochemical constituents present in water extract of Tamarindus indica pulp includes Alkaloids, saponins, steroids, phlobatannins, carbohydrate, terpenoids, phenols, coumarins and leucoanthocyanins. The in vitro cholesterol lowering effect of Tamarindus indica pulp extract shows a positive result on chicken fat, ghee and egg yolk. But in case of pork fat and cod liver oil no beneficial change was observed.