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Growth performances, organ development and blood parameters of rats fed graded levels of steeped and cooked taro tuber (Colocasia esculenta var esculenta) meal

A Teguia, P B Telefo* and R G Fotso*

Département des Sciences Animales, Faculté d'Agronomie et des Sciences Agricoles (FASA), Université de Dschang, BP. 70 Dschang, Cameroun. Tel. (237) 781 11 37.
Email: alexisteguia@yahoo.fr   ;   alexisteguia@justice.com
*Département de Biochimie, Faculté des Sciences, Université de Dschang, BP. 67 Dschang, Cameroun
bphelix@yahoo.co.uk   and   ghislainefotso@yahoo.fr

Abstract

A 42-day study on subchronic toxicity of the steeped and cooked taro meal was conducted using 16 males and 16 females 28-day old rats. Corn meal was substituted in a complete randomized design either at 0, 25, 50, or 100% levels with steeped and cooked taro meal in the usual rats' diet.

The incorporation of steeped and cooked taro meal in the diet didn't have any significant effect (p>0.05) on feed consumption, weight gain, and feed efficiency ratio of rats and on the macroscopic state of organs from tested animals. No significant difference (p>0.05) was recorded between the groups of animals for relative weights of the organs and studied blood parameters. No significant effect (p>0.05) of taro was recorded on total hepatic proteins, total serum cholesterol and alanine aminotransferase. However, total serum proteins and aspartate aminotransferase significantly (p<0.05) decreased with increasing incorporation of steeped and cooked taro meal in the diet of rats. The steeped and cooked taro meal was not found toxic at short-term and could be recommended for animal feeding.

Key words: blood parameters, growth performances, rats, steeped and cooked taro meal


Introduction

Maize is the main cereal involved in animal nutrition. It represents the main energy source and constitutes 60 to 70 % of monogastrics' diets especially pigs and poultry. Worldwide, about 66 % of all maize is used for feeding livestock, 25 % for human consumption and 9 % for industrial purposes (Raemaekers 2001). In tropical countries including Cameroon, there has always been competition between human and animals for maize and the importation has partially solved the problem. However, with the increase demand of maize for feed production; its price on international market is forecast to increase very rapidly. This insufficient cereal production has led to an increasing interest for substitution of maize with other agricultural products in animal.

The studies on the substitution of maize in the finisher diet of poultry with dried leaves of sweet potatoes (Hypomoea batatas) and ndole (Vernonia spp) showed that these leaves could be used efficiently to replace maize in the finisher diet up to 300 g.kg-1 without significantly (p>0.05) affecting the body weight gain, feed consumption and feed efficiency ratio (Teguia et al 1993). Lyayi et al (2001) established the possibility of formulating diets for pigs including cocoa husk pods which, when treated with urea, improved the nutritive value of feed thus resulting in better production performance. Whole cassava plant could also be used to replace 25 to 50 % of maize in the starter diet of broiler chicken without any negative effect on their health but, with 13 to 26 % reduction of feed conversion (Akinfala et al 2002). Leterme et al (2005) studied the chemical composition, the nutritive value and voluntary intake of cocoyam (Xanthosoma sagittifolium) leaves and those of two tropical tree (Trichanthera gigantean, Morusalba) foliage in pigs and it was concluded that low energy density is the main limiting factor of tree foliage for pig nutrition but that they are good sources of minerals and well-balanced proteins. However, performances recorded in all these studies where limited by the low energy content of tested products.

Taro (Colocasia esculenta) has an energy content of above 4000 kcal/kg of dry matter and can be a good source of energy in animal feed. The total production in Africa in 1998 reached about 6.5 millions metric tons representing 75 % of the total world production (8.5 millions tons) (Onwueme 1999). The average taro yield in Africa is about 5.1 t/ha as compared with 1.6 t/ha for maize (Raemaekers 2001). However, in general, taro corm has a poor post harvest storage quality. In Cameroon, the high rate of post harvest loss and lack of proper scientific attention to this problem has been associated with more than 70 % drop in annual production (MINAGRI 1981, MINAGRI 1999). In addition, taro contains oxalates which altered its nutritive value by fixing some minerals such as calcium, zinc, iron that become therefore unavailable to the animal body. Different methods have been developed to eliminate or to least reduce the effect of antinutritionnal factors (ANFs) in animal feed among which, cooking in water, steeping and the combination steeping and cooking in water.

Although taro is consumed by human in Cameroon, no toxicological information on rodent or other species of livestock is available. The objective of this study was therefore to evaluate the subchronic toxicity and potential of steeped and cooked taro meal as a replacement of maize in the diets of laboratory rats.

Materials and methods

Animals and diets

Raw taro tubers brought from the local market were washed, peeled with a sterile knife and cut into slices of about 5 mm diameter. The sliced tubers were steeped in water (1 l/kg tubers) at room temperature for four days, and then cooked in water at about 100° C for one hour. Steeped and cooked tubers were oven-dried at 45° C for 5 days, grounded in a feed mill then stored in hermetically closed container. In the usual diet of rats consisting of 66 % maize flour, 20 % roasted soybean flour, 10 % fish meal, 1.4 % of oyster shell, 1 % cooking salt, 0.5 % cotton oil and 0.1% of biomultivitamins used as control diet, maize flour was replaced with steeped and cooked taro meal at either 25 % (diet 1), 50 % (diet 2) or 100 % (diet 3) level. The experimental chemical characteristics of steeped and cooked taro meal and the calculated chemical characteristics (Sauvant et al 2002)of experimental diets are given in table 1.

Table 1.  Chemical characteristics of steeped and cooked taro meal and experimental diets containing graded level of steeped and cooked taro meal

Diets

Crude protein, g/100 g

Crude cellulose, g/100 g

Crude lipids, g/100 g

Total  ash, g/100 g

Total carbohydrates, g/100 g

Crude energy, kcal/kg

Steeped and cooked taro meal

7,08

5,61

3,93

2,38

86,61

4389,94

Control

18.18

2.96

9.05

5.72

66.98

4234.97

Diet 1

17.95

3.52

9.09

5.91

66.91

4322.41

Diet 2

17.78

4.08

9.13

6.11

66.85

4409.85

Diet 3

17.44

5.21

9.19

6.49

66.72

4584.73

Thirty-two 28 days-old wistar albino rats (16 males and 16 females) were randomly divided into four groups of four males and four females each. The feed and water were provided ad libitum. Group 1 serves as control and was fed the diet without any taro meal (diet 0). Groups 2, 3 and 4 were respectively fed diets 1, 2 and 3 containing respectively 25, 50 and 100 % of steeped and cooked taro meal as a replacement of maize for 42 days.

Daily feed intake and weekly body weight were recorded and weekly feed efficiency ratio calculated. After the 42 experiment days, all the animals were submitted to an eight-hour fasting, anaesthetized with chloroform vapors and bled. Blood samples were collected by cardiac puncture using a sterile syringe. Samples of heart, liver, spleen, lungs, kidney, surrenal glands, ovaries, uteruses, deferent canal, epididymis, seminal vesicle, prostate, testicles, were immediately collected and weighed, their relative weights calculated. The livers samples were frozen at -20° C, and later processed for hepatic protein.

Biochemical and hematological analysis

Serum samples were analyzed for aspartate aminotransaminase (AST), alanine aminotransaminase (ALT) activities using commercial kits as described by Reitman and Frankel 1957) and Bergmeyer (1972), creatinine concentration using commercial kit Creatinine Liquicolor (Germany) as described by Schirmeister (1964). Serum was analyzed for Total protein concentration using Biuret method (Gornal et al 1949) Cholesterol concentration was by using commercial kit established by CHRONOLAB as described by Richmond (1973)  and Flegg (1973). The liver supernatant was analyzed for Total proteins using Biuret method (Gornal et al 1949).

White blood cell (WBC) and red blood cell (RBC) count were determined as described by Theml (2000) while packed cell volume (PCV) was evaluated according to Benson et al (1992).

Statistical analysis

All the data collected during the 6-week study period on weight gain, feed intake, feed efficiency ratio, biochemical and hematological parameters were submitted to a one-way analysis of variance. The Student's t-test was used for the separation at 5% probability level. The JMPIN software version 4 of SAS (2001) was used.

Results

Growth performances and relatives weight organs of rats

Average feed intake and feed efficiency ratio increased with increasing level of taro meal in the diet over the 6-week period (table 2).

Table 2.   Average weekly feed intake (g) of diets, feed efficiency ratio (g feed/g weight gain) and animals growth rate (% increase in weight gain) of wistar albino rats fed graded level of steeped and cooked taro meal

Treatments

Feed intake

Feed efficiency ratio

Growth rate

Control

167.26NS

3.82 NS

203.80 NS

Diet 1

179.83 NS

4.85 NS

170.11 NS

Diet 2

183.40 NS

5.04 NS

165.00 NS

Diet 3

191.07 NS

5.12 NS

177.62 NS

SEM

12.06

1.14

11.17

NS: Mean within the column are statistically comparable (p>0.05).

SEM: Standard error of the mean

In contrary, the higher growth rate was recorded with the control diet. However, no significant difference (p>0.05) was observed among the treatment groups for average feed intake, growth rate and feed efficiency ratio. No mortality was recorded over experimental period.

At killing, no significant difference was observed on the relative weights of collected organs from the different groups of animals (tables 3 and 4).

Table 3.   Average relative weights (weight organ 100/weight animal) of some organs of wistar albinos rats fed graded level of steeped and cooked taro meal

Treatments

Heart

Lungs

Spleen

Liver

Pancreas

Kidneys

Surrenal glands

Control

0.276 NS

0.639 NS

0.216 NS

3.371 NS

0.284 NS

0.551 NS

0.024 NS

Diet 1

0.285 NS

0.639 NS

0.235 NS

3.181 NS

0.120 NS

0.555 NS

0.019 NS

Diet 2

0.293 NS

0.578 NS

0.263 NS

3.320 NS

0.157 NS

0.592 NS

0.028 NS

Diet 3

0.283 NS

0.663 NS

0.224 NS

3.345 NS

0.187 NS

0.628 NS

0.021 NS

SEM

0.011

0.076

0.023

0,172

0,655

0,232

0,039

NS: Means within column are statistically comparable (p>0.05)

SEM: standard error of the mean



Table 4.   Average relative weights (Weight organ 100/weight animal) of reproduction organs of wistar albinos rats fed graded level of steeped and cooked taro meal

Groups

Males

 

Females

Prostate

Seminal vesicle

Epididymis

Testes

Deferent Canal

 

Ovaries

Uterus

Control

0,034 NS

0,302 NS

0,200 NS

0,824 NS

0,035 NS

 

0,058NS

0,208NS

Diet 1

0,048 NS

0,166 NS

0,271 NS

0,879 NS

0,047 NS

 

0,034NS

0,128 NS

Diet 2

0,046 NS

0,212 NS

0,209 NS

0,932 NS

0,044 NS

 

0,048NS

0,219 NS

Diet 3

0,037 NS

0,184 NS

0,208 NS

0,895 NS

0,035 NS

 

0,057NS

0,161NS

SEM

0.009

0.039

0.014

0.040

0.006

 

0.006

0.046

NS: Means within columns are statistically comparable (p>0.05),

SEM: standard error of the mean.

Hematological and biochemical parameters of rats

The PCV percentage and RBC counts tend to decrease with increasing level of steeped and cooked taro meal in the diet while WBC counts tended to increase (table 5).

Table 5.  Average packed cell volume, red blood cells and white blood cells changes in rats fed experimental diets containing steeped and cooked taro flour as a replacement of maize meal

Groups

PCV, %

RBC, 105/mm3

WBC, 102/ mm3

Control

45.88NS

95.08NS

56.75NS

Diet  1

45.00 NS

90.43NS

56.75NS

Diet  2

44.75 NS

89.43NS

57.25NS

Diet  3

43.50 NS

84.21NS

57.75NS

SEM

0.95

4.85

3,92

NS: Means within column are statistically comparable (p>0.05)

SEM: standard error of the mean.

However, no significant (p>0.05) effect of the taro meal in the diet was recorded on PCV percentage, RBC and WBC counts.

The effects of steeped and cooked taro meal on serum parameters are given in table 6.

Table 6.   Concentration of serum and hepatic constituents of rats fed graded levels of steeped and cooked taro meal

Groups

Total hepatic Protein,  mg/g

Total serum protein, mg/ml

Total serum Cholesterol, mg/dl

Serum AST, U/L

Serum ALT, U/L

Serum creatinine, mg/dl

Control

177.00a

116.12a

86.72a

107.35a

57.58a

2.97a

Diet 1

188.87a

90.93b

97.66a

80.57b

38.67a

3.00a

Diet 2

210.25a

81.54b

68.36a

68.58b

39.38a

2.57a

Diet 3

170.77a

78.55b

70,70a

72.47b

45.70a

3.00a

SEM

19.12

3.92

10,16

5.17

3.32

0,37

a, b: Means carrying the same superscript within a column are statistically comparable values (p>0.05),

SEM: standard error of the mean.

No significant differences (p>0.05) were recorded between the treatment groups for ALT activity, total cholesterol, total hepatic protein and creatinine concentration. AST activity and total serum protein concentration were significantly lower (p<0.05) in rats feed diet containing steeped and cooked taro meal as compared with control group.

Discussion

The increase in feed intake and feed efficiency ratio associated with a decrease in growth rate of animals as levels of steeped and cooked taro meal in the diets increased could be due to a poor utilisation of diet containing tested feedstuff. Steeped and cooked taro meal contained 5.61 % of crude fibre as compared with 2.2 % for maize (Sauvant et al 2002). These results agree with reports that, chickens' growth rate decreases with increase of fiber in diet (Agbede 2003). In the same thought, Djopnang (2004) demonstrated that, the increase incorporation level of "karité" flour in rats' diet led to growth delay associated with increase dietary fiber from 1.6 to 3.8 %.

The absence of significant difference among treatment groups for the relative organ weight and hematological parameters added to the absence of mortality suggested that steeped and cooked taro flour was not toxic. An increase in the weight of spleen (Robins 1974), a decrease in that of testes (Bhargava 1984) liver and kidney (Solomon et al 1993) could indeed be an indication of a toxic environment. However, although no significantly (p>0.05), RBC counts decreased while WBC count increased. According to Robins (1974), in a toxic environment, RBC count significantly decreased while WBC increased. The results of the present study could suggest the presence of residual ANFs in steeped and cooked taro meal.

The concentration of creatinine, cholesterol and hepatic proteins varied but not significantly. No clear effect of taro meal on these parameters was observed as inclusion level increased. The significant reduction of total serum proteins with increasing level of taro meal could be associated with the corresponding increase of dietary fibre known to alter bioavailability of dietary proteins. Cossack and Rojhani (1992) reported that excessive dietary fibers disturb digestive enzymes action by binding to minerals and B group vitamins that usually act as enzyme cofactors in proteins metabolism. According to Solomon et al, (1993) and Adam (1998), AST and ALT activities into the blood stream significantly increase in a toxic environment. In rats, 90 % of AST are present in holoenzyme form. During pyridoxine deficiency, the activity rate of these holoenzymes in relation with the total activity is significantly reduced but after a pyridoxine injection, the rate of holoenzyme raises rapidly while the total activity rises after several days (Okada et Kondo 1982, Koruda et al 1982). Koruda et al (1982) demonstrated the formation of an inactive form of AST in liver of pyridoxine deficient rats. In the present study, the activities of these enzymes decreased and this evolution was significant for AST. This fact could probably result from the lack of pyridoxine in the studied taro meal.

Conclusions

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Received 19 January 2007; Accepted 26 March 2007; Published 1 June 2007

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