Introduction

The use of haematological values as indices of the state of animal health is receiving a lot of research effort .The research has been centered on its application as a means of diagnosing disease and stress-induced conditions20 . Many workers have described the use of haematological differential resistancce or otherwise of some animals to variations in diet composition and resultant health implications in our environment118. Blood is involved in distribution of Homones, thus maintaining homeostasis in the body and coordinating activities of various organs of the body. It play important role in immunity and blood clotting21 .

Dietary components have measurable effect on blood constituents. Examination of blood therefore provides a valuable opportunity to clinically investigate the presence of several metabolites and other constituents in the body of animals (Hafkenscheid and Dijt, 1979).

Blood examination is also a good way of assessing the health status of an animal as it plays a vital role in the physiological, nutritional and pathological status of the animal8. Changes or differences in blood composition could be evaluated if normal values are known. Haemotological parameters usually measured are haemoglobin (HB), packed cell volume (PCV), white blood cell (WBC), and red blood cell (RBC) (Hafkenschieid and Dijt, 1979).

Studies have shown that alcohol may benefit many bodily organs, including the heart and the brain. However, the benefits are available only when wine is taken in moderation as over consumption of alcohol including wine can cause some diseases including cirrhosis of the liver and alcoholism13. Alcohol, a major component of wine, is a drug that depresses the central nervous system19. Alcohol has several biochemical effects after consumption, it alters the intracellular NAD+/NADH ratio and this affects the equilibrium constant of a number of important metabolic reactions that utilize these two cofactors16. In view of this effect, these works is aimed at evaluating the toxicological effect of wines produced on haematological indices in rats as compared to imported red wine.

Materials and Methods

Wine Samples

Pineapple wine, Cashew wine and Banana wine were produced in the laboratory of the Department of Microbiology University of Ilorin, Ilorin, Nigeria using Saccharomyces cerevisae as a fermenting organism according to Bisson6. Red wine (Carlo Rossi) imported from USA was used as a control for this work.

Animals and Treatments

Animals weighing an average of 160g were bred and housed in the Animal House of the Department of Chemical science, Ajayi Crowther University, Oyo, Nigeria. They were kept in wire meshed cages and fed with commercial rat chow (Bendel feeds Nigeria ltd) and supply water ad libitum.

Seventy eight (78) albino rats (wistar strain) were divided into five groups:

Group I consist of only 6 rats served as the negative control to which only distilled water (no wine) was administered orally.

Group II consist of 18 rats divided into 3 subgroups A, B, C received 6.25 ml/kg body weight of 5%, 7.5% and 10% alcohol content of the Red wine (positive control) respectively for eighteen days.

Group III consist of 18 rats divided into 3 sub groups A, B, C received 6.25 ml/kg body weight of 5%, 7.5% and 10% alcohol content of the Pineapple wine respectively for eighteen days.

Group IV consist of 18 rats divided into 3 sub groups A, B, C received 6.25 ml/kg body weight of 5%, 7.5% and 10% alcohol content of the Cashew wine respectively for eighteen days.

Group V consist of 18 rats divided into 3 sub groups A, B, C received 6.25 ml/kg body weight of 5%, 7.5% and 10% alcohol content of the Banana wine respectively for eighteen days.

The animals were sacrificed 24 hours after the last treatment.

Collection of Blood Samples for Haematological Determination

The rats were sacrificed by cervical dislocation. Blood samples were collected by occular punctures into EDTA bottles for haematological determination.

Determination of Haematological Parameters

Packed Cell Volume Determination (PCV)

PCV (the volume by percentage of red blood cells in the whole blood) was determined by the method of Baker3.

Determination of Haemoglobin concentration (HB)

Haemoglobin concentration was determined by the method of Agbede1.

Determination of red Blood Cell count

RBC count is the number of RBC in a stated volume of whole blood. It was determined by the improved Neubauer’s counting chamber according to the method of Blaxhall4.

Determination of Total White Blood Cell (WBC) Count

The total white blood cell count was determined by the visual method as described by Dacie and Lewis7.

Differential Leucocytes Counts

Differential Leucocytes Counts was determined by the method of Dacie and Lewis7.

Other red blood cell indices

Other red blood cell indices (MCV, MCH & MCHC) were calculated as described by Jain12.

Statistical analysis

The data were analyzed using one way ANOVA followed by Duncan multivariable post-hoc test for comparison between control and treated rats in all groups. P values less than 0.05 were considered statistically significant.

Results and Discussion

Table 1 shows the effect of different wines of varying alcoholic contents on Red Blood Cell indices, while the effect of different wines of varying alcoholic contents on Differential White Blood Cell Count was presented in Table 2. From the result it can be seen that % PCV and RBC was significantly reduced in those rats treated with 10% pineapple wine and 7.5% and 10% cashew wine respectively, while there was a slight increase in all the banana wine treated group when compared to the red wine (p<0.05). Similarly, Hb content was significantly reduced in all the pineapple and cashew wine treated group, while there was a significant increase in the banana treated group at all the alcohol content when compared to the red wine (p<0.05).

Table 1. Effects of wine samples on Red Blood Cell indices.

Each Value is a Mean of Three Determinations ± SEM.

Values along the same row are not significantly different (P>0.05).

KEY: PCV- Pack cell volume, MCH – Mean cell haemoglobin, HB –Haemoglobin, RBC- Red blood cell, MCV – Mean cell volume, MCHC- Mean cell haemoglobin concentration.

Table 2. Effects of wine samples on Differential White Blood Cell Count.

Each Value is a Mean of Three Determinations ± SEM.

Values along the same row are not significantly different (P>0.05).

KEY: LYM –Lymphocyte EOS – Eosinophil, NEU – Neutrophil TWBC – Total white blood cell.

There was little or no difference in the MCH, MCV and MCHC count in the entire treated group when compared with the red wine. The present investigation has provided information on some haematological effects that have resulted from the administration of some selected locally produced fruit wines to rats. The significant decrease in the RBC count, Hb concentration and PCV level in the blood of the rats fed with pineapple and cashew wine is an indication that a state of mild anaemia had occurred as a result of their treatment.

Kelly14 reported that reduction in amount of Hb below the normal level give rise to a state of mild anemia which may or may not be due to reduction in the RBC population and /or PCV value. As previously reported by Harold and Ballard11, the reductions in RBC count in both pineapple and cashew wine treated group suggests a depression of haematopoietic process. The elevated Hb concentration in the rats fed with banana wine is consistent with increase RBC production in the group. This may be supported by previous reports that protein rich diets increased both haematocrit levels and haemoglobin concentration in human and animal studies2.

Since PCV levels reflect the extent and efficiency of oxygen uptake and transfer to tissue, the low values in the rats treated with pineapple and cashew wine may reflect low oxygen uptake and transfer to tissues, signifying a reduction in the body’s metabolic activity.175, while an increased in PCV level in rats treated with Banana wine compared to red wine indicate a high metabolic activity and good oxygen uptake175.

Also, there was a significant reduction in TWBC count in all the rats fed with different alcohol content of all the wine samples when compared with the red wine. There was significant reduction in percentage Lymphocyte in the rats fed with all wine samples at 7.5% and 10% alcohol content, while there was a significant increase in percentage Neutrophil and percentage Eosinophil in the rats fed with banana wine at different alcohol content when compared with the red wine.

The significant decrease in TWBC count and % Lymphocytes by the Pineapple and Cashew wine suggests that they may be immunosuppressive. The reduction in the TWBC count could be due to reduced production of White blood cells, redistribution of white blood cells from peripheral blood into the tissues or rapid destruction of white blood cells9.

Percentage decrease in Eosinophil and percentage Neutrophill counts observed across the interval of all the treated groups excluding banana group reflect stress increase on the immune system, suggesting a lowering of resistance to infections agents. Since lymphocyte and neutrophill have a major role in fighting foreign organisms.

The effectiveness of banana wine in improving the haematological indices can be attributed to the submission of Kotecha and Babasahde15 that banana is the most nourishing of all fruits; it contains nearly all the essential nutrients, including minerals and vitamins and has several medicinal properties.

Conclusion

It can be concluded from the haematological studies that banana wine is of good quality when compared to cashew and pineapple wine, and its imported Red wine equivalents.