Some biochemical, haematological and histological responses were studied in rats undergoing a long term feeding with a Telfairia occidentalis-supplemented diet. Biochemical and hematological parameters investigated included serum protein, total cholesterol, lipid peroxidation, haemoglobin, white blood cells, Aspartate aminotransferase, Alanine aminotransferase, Alkaline phosphatase, Glutathione-stransferase and Superoxide dismutase. Histological changes associated with Telfairia occidentalissupplemented

diet on the liver, intestine and testes were also examined. Results showed that Telfairia occidentalis-supplemented diet caused a significant increase (p<0.05) in weight and haemoglobin.

Cholesterol and lipid peroxidation were significantly reduced (p<0.05). There were however no significant

changes in the enzymes including Aspartate minotransferase, Alanine aminotransferase, Alkaline phosphatase, Glutathione-s-transferase and Superoxide dismutase. Only Alkaline phosphatase was significantly reduced (p<0.05). Histological changes showed hypertrophy of the intestinal propia and reduced globlet cells while the testes exhibited thick basement membrane and large spermatogonia

Iweala, E. E. J.

Obidoa, O.

English

Covenant University Repository

Pakistan Journal of Nutrition 8 (8): 1199-1203, 2009ISSN 1680-5194© Asian Network for Scientific Information, 20091199Some Biochemical, Haematological and Histological Responses to a Long TermConsumption of Telfairia occidentalis-Supplemented Diet in Rats Emeka, E.J. Iweala*  and Onyechi Obidoa  1   2Department of Biological Sciences, Biochemistry and Molecular Biology Unit, 1Covenant University, P.M.B 1023, Ota, Ogun State, Nigeria Department of Biochemistry, University of Nigeria, Nsukka, Nigeria 2Abstract: Some biochemical, haematological and histological responses were studied in rats undergoinga long term feeding with a Telfairia occidentalis-supplemented diet. Biochemical and hematologicalparameters investigated included serum protein, total cholesterol, lipid peroxidation, haemoglobin, whiteblood cells, Aspartate aminotransferase, Alanine aminotransferase, Alkaline phosphatase, Glutathione-s-transferase and Superoxide dismutase. Histological changes associated with Telfairia occidentalis-supplemented diet on the liver, intestine and testes were also examined. Results showed that Telfairiaoccidentalis-supplemented diet caused a significant increase (p<0.05) in weight and haemoglobin.Cholesterol and lipid peroxidation were significantly reduced (p<0.05). There were however no significantchanges in the enzymes including Aspartate aminotransferase, Alanine aminotransferase, Alkalinephosphatase, Glutathione-s-transferase and Superoxide dismutase. Only Alkaline phosphatase wassignificantly reduced (p<0.05). Histological changes showed hypertrophy of the intestinal propia and reducedgloblet cells while the testes exhibited thick basement membrane and large spermatogonia.Key words: Telfairia occidentalis, weight, haemoglobin, cholesterol, lipid peroxidationINTRODUCTIONRegular consumption of plant foods are associated withnumerous health benefits rooted in their variousphysiological effects as a result of their phytochemicaland nutritional constituents (Hunter and Fletcher, 2002).Green leafy vegetables are particularly important inpromoting health because of their rich sources of MATERIALS AND METHODSnutrients (Gupta and Prakash, 2009). Telfairia Preparation of leaves of T. occidentalis: Fresh leavesoccidentalis (Fluted pumpkin) is a common tropical of T. occidentalis were harvested fresh after 4 weeks ofgreen leafy vegetable native to many African countries leaf emergence from a garden farm in South-Easternespecially Eastern Nigeria (Burkett, 1968). It thrives in Nigeria. The leaves were picked, air-dried and ground tohumid climate and well drained soils and is usually a coarse powdered form with a hammermill. Thecultivated in garden and family farms around homes. proximate composition of the ground leaves of T.Telfairia occidentalis belongs to the family occidentalis were determined according to theCurcubitaceae and has simple, dark green veined procedure of AOAC (1975) in order to formulate anleaves that is as wide as 18 cm and long as 35 cm. T. isonitrogenous and isocaloric feed together with otheroccidentalis contains nutrients such as proteins, feedstuff.carbohydrates, vitamins, minerals and fiber (Fasuyi,2006). It also contains oxalates, saponins, glycosides, Formulation of experimental feeds: Two feeds namelyflavonoids, alkaloids and resins (Tindall, 1968; Akubue Control Diet (CD) and Telfairia Occidentalis-et al., 1980). The nutritional content of T. occidentalis supplemented Diet (TOSD) were formulated. Themakes it desirable as dietary supplement for humans. feedstuff used in formulation of feed included feed gradeThe leaves are widely consumed and used in maize, corn flour, fish meal. groundnut meal, bone mealpreparation of soups and salads. In ethno medicine, the and vitamin premix purchased from a reputablefresh leaves are used in the treatment of anemia, manufacturer (Jocan Agro Ltd, Umuahia, Nigeria). Theconvulsion and malaria (Alada, 2000; Gbile, 1986). The proximate composition of the leaves of T. occidentaliswide spread consumption of leaves of T. occidentalis was used as a guide in the formulation of thereflects a substantive measure of its nutritional experimental diet that was both isocaloric andacceptability of as a health promoting plant food. Inspite isonitrogenous. The Control Diet (CD) was formulatedof the widespread use of T. occidentalis, there is scanty without the inclusion of T. occidentalis leaf powder whileinformation on its various biological and histologicaleffects. This study investigated some biochemical,haematological and histological changes associatedwith the long term consumption of a diet supplementedwith leaves of T. occidentalis in albino rats.Pak. J. Nutr., 8 (8): 1199-1203, 20091200the Telfairia Occidentalis-supplemented Diet (TOSD) Enzyme assays: Glutathione-s-transferase waswas incorporated with 5% of T.occidentalis leaf powder.Animal grouping and experimental design: A total oftwenty albino male rats of the Wistar stock aged fourweeks old used for the experiment were purchased fromthe animal house of the department of Biochemistry,University of Nigeria, Nsukka. The animals had anaverage weight of 45.5±2.85 g. The animals were fedwith Control Diet (CD) at the start of the experiment foracclimatization and given water ad libitum.The experimental design was completely randomizedinvolving random distribution of the rats into twoexperimental groups namely Control Group (CG) andExperimental Group (EG) of ten animals each havingsimilar group mean weights at the start of theexperiment. The animals in CG and EG were fed ControlDiet (CD) and Telfairia Occidentalis-supplemented Diet(TOSD) respectively and given water ad libitum for 180days (9 6 months). Weight changes of animals in thetwo groups were recorded monthly.Collection of blood, organs and tissue samples: At theend of the 180 days of feeding, six animals from eachgroup were anesthetized and decapitated. Bloodsamples were collected in labeled sample bottles withdrops of Ethylenediaminetetraacetic Acid (EDTA). Serumsamples were collected in sample bottles without EDTA,allowed to clot and centrifuged at 5000 rpm for 10 min.The liver, intestines and testes were promptly excisedsoon after decapitation and stored in 10% formyl saline.Haematological studies: Hemoglobin and White BloodCells (WBC) were determined according to methodsdescribed by Dacie (1975) and Dacie and Lewis (1991).Biochemical determinations: Total serum protein wasdetermined by the Lowry method (1951), total and freecholesterol were determined by the method of Searcyand Bergquist (1960). Lipid peroxidation wasdetermined by the method of Walin et al. (1993).determined according to the method of Habig et al.(1974). Superoxide dismutase was assayed by themethod of Jewett and Rocklin (1993). AspartateAminotransferase (AST), Alanine aminotransferase(ALT) and Alkaline Phosphatise (ALP) were determinedusing Randox test kits.Histological studies: Histological studies on the organsand tissues were done according to proceduresdescribed by Disbrey and Rack (1974) and Drury andWellington (1967). The tissues were viewed under lightmicroscopy.Statistical analysis: Results were expressed as Mean± Standard Error of Mean (SEM) for triplicatedeterminations. A One-way analysis of variance (ANOVA)for a completely randomized design and Duncan’smultiple range tests were used to analyze experimentaldata. Values were considered significant at p<0.05.RESULTSThe results of the long term effect of feeding TelfairiaOccidentalis-supplemented Diet (TOSD) on weight andsome biochemical parameters are presented in Table1. TOSD caused a significant percentage increase(p<0.05) in weight gain when compared to the control.There were significant reductions in the cholesterol leveland lipid peroxidation to 0.88±0.10 Mm/L and 3.05±0.30respectively in animals fed TOSD. Hemoglobin levelswere also remarkably increased in the animals fed withTOSD. The changes in serum protein and WBC countwere not statistically significant.The effect of TOSD on some enzyme levels arepresented in Table 2. All the enzymes did not show anysignificant changes except Alkaline phosphatase whichsignificantly decreased to 76.3±3.68.The histological features of liver, intestine and testes ofthe rats fed with Control Diet (CD) and TOSD are shownin Fig. 1a, 1b, 2a, 2b, 3a and 3b. There was a slightdistortion in the hepatic architecture and plates of theanimals fed with TOSD. Also the intestinal propia wereTable 1: Effect of T.occidentalis-supplemented diet (TOSD) on some biochemical and haematological parameters (n = 3)LipidProtein Cholesterol Hemoglobin WBC PeroxidationExperimental groups (Diet) Weight (%) (mg/ml) (mM/L) (g/dl) (10 /mm ) (mg/ml)3 3Control diet (CD) 60.0±7.28 261±16.4 1.43±0.18 9.8±0.50 5.13±0.29 5.03±0.23T.occidentalis- 70.8±6.82* 279±13.2 0.88±0.10* 12.8±0.95* 5.27±0.17 3.05±0.30supplemented diet (TOSD)**p<0.05Table 2: Effect of T.occidentalis-supplemented diet (TOSD) on some enzymes (n = 3)Experimental groups (Diet) GST (IU/l) SOD (U/mg protein) AST (IU/l) ALT (IU/l) ALP (IU/l)Control diet (CD) 5.41 ± 0.82 70.7 ± 3.68 88.7 ± 7.41 45.3 ± 5.3 88.6 ± 8.96T.occidentalis- 7.13 ± 1.44 59.7 ± 2.62 63.0 ± 3.56 32.3 ± 4.03 76.3 ±3.68supplemented diet (TOSD)**p<0.05Pak. J. Nutr., 8 (8): 1199-1203, 20091201Fig. 1a: Photo micrography of a cross section of liver of Fig. 2a: Photo micrography of a cross section ofrats fed with Control Diet (CD). MAG. x 40 intestine of rats fed with control diet (CD). MAG.Fig. 1b: Photo micrography of a cross section of liver ofrats fed with Telfairia occidentalis- Fig. 2b: Photo micrography of a cross section ofsupplemented Diet (TOSD). MAG. x 40 intestine of rats fed with Telfairia occidentalis-hypertrophied and while the globlet cells were reducedin the group fed TOSD. The testes of the rats fed withTOSD exhibited thickened basement membrane andlarge spermatogonia when compared to the control.DISCUSSIONTelfairia occidentalis possesses unique nutritional andphytochemical properties which can have variedphysiological and biochemical effects (Longe et al.,1983). In this study, the long term feeding of T.occidentalis-supplemented diet (TOSD) caused asignificant increase in weight of the animals which maybe due to its content of rich nutrients. Telfairiaoccidentalis contains nutrients such as amino acids,fatty acids, mineral and vitamins (Fagbemi, 2007). Theincrease in weight may also be related to increased feedintake. Leaves of T. occidentalis are highly palatable andconsumed largely in a variety of diets in South-EasternNigeria. The feed may also have effect on the feedconversion  rate  of  the  animals (Adedapo et al., 2008).x 40supplemented diet (TOSD). MAG. x 40 In addition certain phytochemicals found inT.occidentalis may affect intake and digestibility ofnutrients (Dube et al., 2001). Lipid peroxidation wassignificantly reduced (p<0.05) in the animals fed withTOSD. This result confirms the antioxidant effects ofTelfairia occidentalis extracts reported by Nwanna andOboh (2007).Anti oxidant effects of vegetables such asTelfairia occidentalis are attributed to the presence ofphenolic compounds such as flavonoids, polyphenolsas well as vitamins (Halliwell and Gutteridge, 1999). Thenon significant changes in the enzymes namely Alanineamino transaminase (ALT), Aspartate Transaminase(AST), Glutathione-s-transferase (GST) suggests somelevel of hepatoprotective effects of TOSD on the rats(Oboth, 2005). This presumption is also accentuated bythe non significant change in serum proteins which arelargely produced in the liver. Also the non-significantchange in Superoxide Dismutase (SOD) complementsthe decrease in lipid peroxidation previously discussed.Pak. J. Nutr., 8 (8): 1199-1203, 20091202Fig. 3a: Photo micrography of a cross section of testesof rats fed with Control Diet (CD). MAG. x 8Fig. 3b: Photo micrography of a cross section of testesof rats fed with Telfairia occidentalis-supplemented diet (TOSD). MAG. x 8These antioxidant effects of TOSD is indispensable inprotecting the animals that consume the its leavesagainst reactive oxygen species. TOSD significantlyincreased the haemoglobin concentration of the rats.This finding is related to the presence of somehaematological factors including proteins, iron,thiamine, riboflavin and nicotinamide in the leaves ofT.occidentalis (Ganong, 1997; Fasuyi and Nonyerem,2007). This result agrees with the study of Alada (2000)and lends validity to the use of leaves of T.occidentalis intreatment of anemia (Gbile, 1986). The TOSD showedvarious cellular effects on the histological characteristicsof the tissues examined. The liver of rats fed TOSDshowed some irregularity possibly due to the presenceof minute quantities of alkaloids in the leaves ofT.occidentalis. The intestinal histology showed somelevel of atrophy suggesting indigestibility. This could beattributed to the presence of tannins and saponins inleaves of T.occidentalis known to cause indigestion ingrazing animals (Reed, 1995). This observation couldhave toxic consequences. A further study on the effect ofconsumption of TOSD on intestinal function of rats isbeing carried out in our laboratory. Large spermatogoniaseen in the histological examination of the testis of ratsfed with TOSD is related to increase inspermatogenesis. This observation corroborates thefinding of Nwangwa et al. (2007) which showed thatT.occidentalis has regenerative effect on the histology ofrat testes. There are unverified claims by traditionalherbalists in Nigeria that consumption of leaves ofT.occidentalis increases sperm count and reproductivity.REFERENCESAdedapo, A.A., M.O. Sofidiya, M.J. Masika and A.J.Afolayan, 2008. Safety Evaluations of the aqueousextract of Acacia karroo stem bark in rats and mice.Rec. Nat. Prod., 2: 128-134.Akubue, P.I, A. Kar and F.N. Nnachetta, 1980. Toxicity ofextracts of roots and leaves of Telfairia occidentalis.Planta Med., 38: 339-343.Alada, A.R.A., 2000. The haematological effect ofTelfairia occidentalis diet preparation. Afr. 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Some Biochemical, Haematological and Histological Responses to a Long Term Consumption of Telfairia occidentalis-Supplemented Diet in Rats

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Some Biochemical, Haematological and Histological Responses to a Long Term Consumption of Telfairia occidentalis-Supplemented Diet in Rats

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