2. BY *Nkwonta, C. G and **Ezeokonkwo, C. A. *S-E Zonal Biotech. Centre, U.N.N **Department of Biochemistry, U.N.N. December 14, 2010 COMPARATIVE STUDY ON THE BIOCHEMICAL EFFECTS OF MOIST AND DRY HEAT- PROCESSED Tetracarpidium conophorum- BASED DIET IN MALE WISTER RATS.

3. INTRODUCTION

4. Tetracarpidium conophorum Mull.-Arg Euphorbiaceae also known as Plukenetia conophora Mull.-Arg is commonly called the conophor plant and African walnut. Tetracarpidium conophorum Mull.-Arg Euphorbiaceae also known as Plukenetia conophora Mull.-Arg is commonly called the conophor plant and African walnut. The plant is a perennial climber and the nuts are basically harvested and eaten boiled or roasted. The plant is a perennial climber and the nuts are basically harvested and eaten boiled or roasted. It is called ukpa by the Igbos and asala/awusa by the yourbas. It is called ukpa by the Igbos and asala/awusa by the yourbas. Several researchers have reported high nutrient potentials of the plant. Several researchers have reported high nutrient potentials of the plant. The impact of traditional processing on the nutrients and sensory qualities of the nut have also been reported The impact of traditional processing on the nutrients and sensory qualities of the nut have also been reported

5. Figure 1. Tetracarpidium conophorum pods

6. Figure 2. Tetracarpidium conophorum nut exposed in their pods

7. Figure 3. Tetracarpidium conophorum nuts outside the pods.

8. USES OF CONOPHOR NUT Food/ Snacks Food/ Snacks Fertility agent for males Fertility agent for males Leaves are used for treatment of dysentery and other digestive disorders Leaves are used for treatment of dysentery and other digestive disorders Extracts has shown antimicrobial and antifungal activities on Pseudomonas aeruginosa, Candida albicans and Aspergillus niger. Extracts has shown antimicrobial and antifungal activities on Pseudomonas aeruginosa, Candida albicans and Aspergillus niger. The oil is Industrially used in formulation of wood vanish, and paints. The oil is Industrially used in formulation of wood vanish, and paints.

9. Processing food involves subjecting it to controlled conditions which helps to improve its safety, shelf life and to transform certain properties; however, it can produce desirable and undesirable effects Processing food involves subjecting it to controlled conditions which helps to improve its safety, shelf life and to transform certain properties; however, it can produce desirable and undesirable effects Heating generally is known to significantly reduce the effects of antinutrients and other factors which are not necessarily antinutrients in the food substances. It also destroys microbes and inactivates enzymes and toxins. Heating generally is known to significantly reduce the effects of antinutrients and other factors which are not necessarily antinutrients in the food substances. It also destroys microbes and inactivates enzymes and toxins. FOOD PROCESSING

10. In the light of the increased interest in the exploitation of common oil seeds and search for alternatives for animal proteins, it has become very imperative to assess the nutritional and biochemical properties of processed Tetracarpidium conophorum. The knowledge of its potentials in growth and maintenance of animal tissues will aid the nutritionist in making proper recommendations using conphor-based diets In the light of the increased interest in the exploitation of common oil seeds and search for alternatives for animal proteins, it has become very imperative to assess the nutritional and biochemical properties of processed Tetracarpidium conophorum. The knowledge of its potentials in growth and maintenance of animal tissues will aid the nutritionist in making proper recommendations using conphor-based diets Rationale

11. To investigate and compare the biochemical effects of roasted and boiled conophor nut in relation with growth and maintenance of animal tissues using rats. To investigate and compare the biochemical effects of roasted and boiled conophor nut in relation with growth and maintenance of animal tissues using rats. Objective

12. MATERIALS AND METHODS

13. PLANT MATERIALS SAMPLE COLLECTION Fresh and mature conophor nuts were purchased from Ibeagwa market, a small market near the University of Nigeria, Nsukka. The fresh dehusked nuts were roasted in hot sand (100 0 C) for 40 minutes with constant stirring. The nuts were shelled, macerated and dried at 60ºC after which they were ground and stored with freezer bags at 4ºC until use. Fresh and mature conophor nuts were purchased from Ibeagwa market, a small market near the University of Nigeria, Nsukka. The fresh dehusked nuts were roasted in hot sand (100 0 C) for 40 minutes with constant stirring. The nuts were shelled, macerated and dried at 60ºC after which they were ground and stored with freezer bags at 4ºC until use. The dehusked nuts were boiled for 2 hours in a boiling water bath, macerated and dried in a hot air oven at 60ºC. The dried samples were ground and stored with freezer bags in a refrigerator at 4ºC until use. The dehusked nuts were boiled for 2 hours in a boiling water bath, macerated and dried in a hot air oven at 60ºC. The dried samples were ground and stored with freezer bags in a refrigerator at 4ºC until use.

14. PROXIMATE ANALYSIS The proximate composition analysis of roasted and boiled T. conophorum nut were carried out using the standard procedures of AOAC by micro-Kjeldhal method and crude protein content calculated as N × 6.25 The proximate composition analysis of roasted and boiled T. conophorum nut were carried out using the standard procedures of AOAC by micro-Kjeldhal method and crude protein content calculated as N × 6.25

15. DIET FORMULATION The diets were formulated using AIN 93G method for growth phase In Laboratory rats. Three main diets were used- Caesin based diet (CAS) which served as the control, the roasted T. conophorum diet (RTC) and the boiled T. conophorum (BTC). The diets were composed to provide 10% protein. The diets were formulated using AIN 93G method for growth phase In Laboratory rats. Three main diets were used- Caesin based diet (CAS) which served as the control, the roasted T. conophorum diet (RTC) and the boiled T. conophorum (BTC). The diets were composed to provide 10% protein.

16. Table 1 Diets based on caesin, BTC and RTC nut flours as sources of protein. Diet composition (g/kg) DIET INGREDIENT DIET GROUPS CAS RTC BTC CAS RTC BTC Casein114.4-- RTC-415.22- BTC--368.24 Cornstarch466.469225.900263.397 Sucrose116.61756.48065.849 Fibre50.050.050.0 Soy Bean Oil 70.070.070.0 Mineral mix 35.035.035.0 Vitamin Mix 10.010.010.0 L-cystein3.03.03.0 Choline bitartrate 2.52.52.5 BHT0.0140.0140.014

17. EXPERIMENTAL DESIGN Eighteen weanling male albino rats supplied by the Veterinary Medicine Department of the University of Nigeria, Nsukka were divided into three groups of six rats each Eighteen weanling male albino rats supplied by the Veterinary Medicine Department of the University of Nigeria, Nsukka were divided into three groups of six rats each Group 1- Caesin (CAS) Group 1- Caesin (CAS) Group 2- Roasted T. Conophorum (RTC) Group 2- Roasted T. Conophorum (RTC) Group 3- boiled T. conophorum (BTC) Group 3- boiled T. conophorum (BTC) They were housed in individual metabolic cages equipped to separate faeces and urine. They were housed in individual metabolic cages equipped to separate faeces and urine.

18. After a six day adjustment period, the rats were weighed prior to access to the test diets. 15g diet /rat /day were fed to the test group and to the casein group for 35days After a six day adjustment period, the rats were weighed prior to access to the test diets. 15g diet /rat /day were fed to the test group and to the casein group for 35days There was daily urinary collection throughout the last seven days of the feeding period. There was daily urinary collection throughout the last seven days of the feeding period. The rats were sacrificed on the 36th day, blood samples were collected for the biochemical analysis using the occular method. The rats were sacrificed on the 36th day, blood samples were collected for the biochemical analysis using the occular method.

19. ASSAYS Liver function enzymes and renal function parameters were determined using RANDOX TEST KITS Liver function enzymes and renal function parameters were determined using RANDOX TEST KITS Haematological parameters- RBS, PCV, TWBC and Differential WBC were determined standard clinical methods. Haematological parameters- RBS, PCV, TWBC and Differential WBC were determined standard clinical methods. Electrolyte assays (K, Na and Cl) were carried using TECO DIAGNOSTIC KITS Electrolyte assays (K, Na and Cl) were carried using TECO DIAGNOSTIC KITS

20. RESULTS and DISCUSSION

21. PROXIMATE COMPOSITION

22. ALT and AST

23. TB and DB

24. CREATININE 0 0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 CASBTCRTC Concentration (mg/dl) CAS BTC RTC CREATININE LEVELS IN RATS FED CAS, BTC AND RTC DIETS

25. UREA 0 2 4 6 8 10 12 14 CASBTCRTC UREA LEVELS IN RATS FED CAS, BTC, RTC DIETS CONCENTRATION (mg/dl) CAS BTC RTC

26. ELECTROLYTES 0 20 40 60 80 100 120 140 160 180 POTASSIUMSODIUMCHLORIDE ELECTROLYTE LEVELS IN RATS FED CAS, BTC, AND RTC DIETS CONCENTRATION (mEq/L) CAS BTC RTC

27. PCV

28. RBS

29. DIFFERENTIAL WBC COUNT 0 20 40 60 80 100 120 CASBTCRTC DIFFERENTIAL WHITE BLOOD CELL COUNT OF RATS FED CAS, BTC, AND RTC PERCENTAGE (%) NEUTROPHILS LYMPHOCYTES MONOCYTES

30. TOTAL WBC

31. CONCLUSION The normal levels bioassay parameters seen suggests that T. conophorum may contain little or no food toxicant. As seen by the results, roasted T. conophorum diet compared more favourbly with the control than the BTC diet although the differences in general were not quite significant. Both test diets has high food potentials that could be harnessed as source of dietary protein and their wider use as a major contributor to our diets is recommended The normal levels bioassay parameters seen suggests that T. conophorum may contain little or no food toxicant. As seen by the results, roasted T. conophorum diet compared more favourbly with the control than the BTC diet although the differences in general were not quite significant. Both test diets has high food potentials that could be harnessed as source of dietary protein and their wider use as a major contributor to our diets is recommended

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