Rapeseed is commonly known as mustered, Brassica spp in Bangladesh, is an annual, winter season crop a native to temperature regions of Europe (Bhuiyan et al., 2008). Rape is a Latin word means turnip and rapeseed belong to the family Cruciferae is an important oil crop and currently it is the principal oil crop of Bangladesh. It contains 40-45% oil and 20- 25% protein in seed (Mosyakin, 1996; Sarker et al., 2019). Beneficial health effects also arise from the content of phenolic compounds, which are known to exert antimutagenic, anti-inflammatory activities (Cao

et al., 2008; Vuorela et al., 2005). The antioxidant potential of phenolic compounds from rapeseed oil has been reported in the literature (Thiyam et al., 2006; Wakamatsu et al., 2005). This oil is a major source of vegetable oil in the world and contains many active biological compounds such as phytosterols, tocopherol, phenolics, β-carotene (Siger, 2017).

In Bangladesh, rapeseed and mustard are major sources of edible oils and are mostly grown on the residual soil moisture in Rabi season. Many   different

parts of plants may yield oil in actual commercial practice oil is extracted primarily from the seeds of oilseed plants. Vegetable fats and oils are not only edible but also non-edible such as processed linseed oil and castor oil used in lubricants, paints, cosmetics, pharmaceuticals, and other industrial purposes. This fats and oils are used to describe concentrated energy sources found in many foods. Without fats and oils, our bodies would not function properly. The oil Rapeseed is one of the most valuable vegetable fats, due to its nutritional properties. It is a rich source of mono- and polyunsaturated fatty acids, while saturated fatty acids are minor constituents (Gunstone, 2011). The rapeseed oil extracted from the rapeseed top-quality edible oil, which contains rich acids, fast-soluble vitamins, phospholipids, and pigments but lacks cholesterol (Martin Hernandez, 2008). It also contains high- quality oil (38-46%) and protein (20-30%) as well as some antinutritional compounds, such as glucosinolates, phenols, and phytic acid (Zhang, 2007). Rapeseed processing in the food and chemical industry results in some by-products which can be used as animal feeds or as a protein component of the diet (Lomascolo et al., 2012).

Vegetables fats and oils are derived from plants that are composed of mainly triglycerides. Rapeseed oil contains high levels of erucic acid that is damaging to the cardiac muscle of animals, and made it less nutritious in animal feed (Obrien, 2008). It helps to stay away from coronary heart disease and provide energy and transport fat-soluble vitamins A, D, E, and

K. It also provides the essential fatty acids omega 3, 6 and 9 involved in many important functions in the body, including heart health. Help keep us warm and protect internal organs like the kidneys contain plant sterols (McKevith, 2005). However, the particular role of the component of these rapeseeds oil remains yet to be studied (Obrien, 2008). Therefore, the present study was undertaken with the aims and objectives is to determine the antimicrobial activities of these rapeseeds oil against some test pathogens to determine the body weight gain (growth rate) of rats fed with both oil supplements, to measure the amount of food consumed by rats, to calculate Food Efficiency Ratio (FER) of rats fed with rapeseeds oil, and to investigate the adverse condition of rats heart and  liver  by measuring the biomarker  SGOT (AST),

SGPT (ALT), CK-MB and ALP.

2.1 Collection of Seeds - At first fresh, healthy and good quality Mustard (hybrid) seeds collected from BADC Amjhupi, Maherpur and Rai (hybrid) seeds collected from the farmers of “Shehala” Village, Daulatpur, Kushtia. Then the seeds were stored at 4oC in the refrigerator with the sealed plastic packet to avoid the microbial contamination.

2.2 Preparation and Storage of Oil - To preparation of the experimental oil, stored seeds were taken out from the refrigerator to the ghanni mil at Sheikhpara, Jhenidah. The bottle was also stored at -28oC to prevent photo-oxidation. The bottle was preserved in a dark, cool and dry place.

2.3 Media

2.3.1 Nutrient Broth Media - The Nutrient broth is typically made of a powder beef or yeast extract that contains peptones (broken down proteins). The powder is dissolved in water, put in the conical flask and sterilized and incubated for 24 hours or more to encourage bacterial growth.

2.3.2 Nutrient Agar Media - Nutrient agar medium used for the cultivation of microbes supporting the growth of a wide range of non-fastidious organisms.

2.4 Sample Used - Sample used for this purpose are four types of rapeseed oil. Oil samples are Mustard (wild) oil, Mustard (hybrid) oil, Rai (wild) oil, Rai (hybrid) oil. Oil directly extracted from seeds from an oil mill in a local market. Seeds are collected from farmers and local market in Kushtia, a general town in Bangladesh.

2.5 Preparation of Pure Culture and Sample Inoculation - To determine anti-microbial activity of oils, we had to prepare media for bacterial growth. Our experiment is divided into two parts. The first part includes the preparation of pure culture of each bacterial strain and the second part includes the  spread of microbes in an agar plate and inoculate sample.

2.6 Disk Preparation - Make disks to soak sample oil by filter paper in the desired size. Take all disks in a bicker. Seal the bicker with aluminum foil paper to avoid contamination. Then sterilize the disk in the autoclave at 121 oC for 30 min and open the bicker in front of laminar airflow.

2.7 Animal Care and Treatment

2.7.1 Experimental Animals (Wistar rat) - A rat of the species Rattus norvegicus which is bred and kept for scientific research is being used in the laboratory.

2.7.2 Animal Maintenance - Adult healthy male Wistar rats were purchased from animal house, Dept. of Pharmacy, Jahangirnagar University, Savar, Dhaka. With an average weight of about 80-100gm (4 weeks of rat). The rats were housed in a wooden cage with steel wire tops and wood-cob bedding (4 rats per cage) and had access to food and water at the BTGE laboratory.

2.7.3 Diet and Dosing of Oil - At first, the rats were adapted for five days. They were fed standard diet. The standard diet was purchased from animal house, Jahangirnagar University, Dhaka, Bangladesh. The a standard diet consisted of 7% protein source, 20% rice polish, 30% wheat bran and flour, 1.5% soybean oil and common salt, 0.5% vitamin mixture and 2.5% molasses. Each rat was fed about 15gm of diet/day. The appropriate doses of rapeseeds oil and diet given to the rats were carefully weighed using the electrical balance. Feeding was continued for 8 weeks. Amount of food consumed and the individual bodyweight of the rats of group A, B and C ware 89.33, 83.25 and 86.75g respectively.

2.7.4 Calculation of Food Efficiency Ratio (FER) - During the study, the weight of each rat was measured every day using the balance. The food efficiency ratio for each rat was calculated using the following formula: FER=Body weight gain/Food consume.

2.7.5 Sample Collection and Preservation - After 56 days of feeding, the 14 hours fasted rats were subjected to light anesthesia using chloroform. At first, the abdomen was washed using ethanol to avoid contamination. With the use of scissors and wearing gloves the abdomen was opened, blood samples were collected with 3 ml syringe from abdominal aorta or heart. Then the blood sample was transferred into a 1.5ml Eppendrof tube. For coagulation, blood was kept about 10 minutes at room temperature. After centrifugation at 3000 r.p.m for 15 minutes at 4 oC using Thermo scientific centrifuge, then the serum was carefully drawn off and placed in a 1.5 ml Eppendrof tube and preserved at -80 oC until the experiments were performed. Various organs namely heart, liver, kidney, tail, adipose tissue and skeletal muscle were collected, put onto Petridis and washed in 0.9% saline and then

preserved. Heart sample was collected and preserved in Bouins Fluid. The liver, kidney, tail, adipose tissue and skeletal muscle were preserved in formaldehyde for further investigation. Bouins fluid contains 71.43% saturated picric acid, 4.76% glacial acetic acid, and 23.81% formaldehyde, which is known as a strong preservative.

2.8 Biochemical Analysis of Serum

2.8.1 Determination of Serum Glutamate Oxalo-acetate Transaminase SGOT (AST) and Serum Glutamate Pyruvate Transaminase (SGPT) - SGOT and SGPT also have known as aspartate amino-transferase (AST) and alanine amino transferase (ALT), respectively. For the analysis of blood serum SGOT (AST) and SGPT (ALT) is used as a biomarker. Commercially available kits were used for the determination of SGOT and SGPT level in blood serum according to the Optimized UV-Test (Walter et al 1991). For the determination of SGOT and SGPT enzyme reagent (R1) and substrate liquid buffer (R2) were supplied ready-to-use. Prepared working reagent in the ratio of 4 parts enzyme reagent (R1) to 1 part substrate (R2). 

At first, we were taken zero spectrophotometers at 340 nm with distilled water. For each sample and control, 200µl serum sample added with 1000µl mono reagent (R1+R2). Gently mixed and absorbance was recorded after 1 minute at 25º C. Incubation continued at 25ᵒ C and absorbance were recorded again at 2 and 3 minutes. Determined the average absorbance per minute (ΔA/min) and multiplied by the factor 952 for results in U/L.

2.8.2 Determination of Alkaline Phosphatase (ALP)- Kits were used in the determination of alkaline phosphatase according to the Optimized Standard Method (Weber et al., 1998). For the determination of ALP, alkaline phosphatase buffer and substrate liquid buffer were ready to use. Prepare working reagent in the ratio of 4 parts Buffer (R1) to 1 part Enzyme (R2) (i.e. 24 ml Buffer and 6 ml Substrate). At first, we  were taken zero spectrophotometers at 405 nm with distilled water. For each sample and control, 20µl serum sample added with 1000µl mono reagent (R1+R2). Gently mixed and absorbance was recorded after1minute at 37º C. Incubation continued at 37ᵒ C and absorbances were recorded again at 2 and 3 minutes. Then determined the average absorbance per minute (ΔA/min) and multiplied by the factor 2757 for results in U/L.

2.8.3 Determination of Creatine Kinase (CK-MB) - For the quantitative determination of Creatine Kinase-MB in serum manual procedure was used. The procedures given by the manufacturers were followed without any modifications for the determination of these parameters. For the quantitative determination of Creatine Kinase MB (CK-MB) liquid buffer (R1) and enzyme (R2) liquid reagents are supplied ready-to-use. Prepare working reagent in the ratio of 4 parts Buffer (R1) to 1 part Enzyme (R2). At first, we were taken zero spectrophotometers at 340 nm with distilled water. For each sample and control, 1.0 ml working reagent added to cuvet and warmed to 37º for 4 minutes. Added 40 µl of serum to its respective tube and mixed gently. Incubation continued at 37ᵒ C and absorbance were recorded again at 6, 7, 8 and 9 minutes. Then determined the average absorbance per minute (ΔA/min) and multiplied by the factor 8360 for results in U/L.