Nutraceutical Effects of Fermented Whey on the Intestinal and Immune System of Healthy Albino Rats

MATERIALS AND METHODS

Fresh whey sample was obtained from local whey manufacturing point at Akure-Owo express road, Akure. The fresh samples were collected in separate sterile container and were transported immediately to the laboratory for investigation.

Twenty four Wister strain albino rats (Rattus novegicus) aged between 6-8 weeks old used for the investigation was purchased from Biochemistry Department, University of Ilorin, Kwara State. They were divided into groups of four rats per group. During the period of the experiment the rats were kept on water and growers mash. The feeding experiment was performed at the animal house of the Department of Animal Production and Health, Federal University of Technology, Akure. A seven days acclimatization period was observed before the commencement of the experiments.

Effect of Feeding Healthy Rats with Whey on their Performance
The initial weights of rats, weight gained within five days and the final weight gains were recorded for healthy rats. The groups of rat were fed with different volumes of fermented whey (0.5, 1.0, 1.5, 2.0 and 2.5 mL) along with the mash. The rats were killed by cervical dislocation and their blood samples were collected into EDTA bottles for haematological parameters and serum enzymes analyses.

Analysis of Serum Enzymes
Serum Aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were assayed by the method of Reitman and Frankel. The Bessy Lowry and Brock alkali phosphate method was used for the analysis of serum alkaline phosphase.

RESULTS AND DISCUSSION

The results of the haematological studies shown in Table 2 revealed that whey improved the health condition of the infected rats by boosting their immunity. This was evidence by the significant (p<0.05) increase recorded in the values of Packed Cell Volume (PCV) and White Blood Cell (WBC) levels of the healthy rats dosed with different volumes of whey.

Table 1:	Haematological parameters of healthy albino rats before whey administration
Control: Rats without whey. PCV: Packed cell volume; WBC: White blood cell. Values are mean of four replicates±SD. Values followed by similar alphabets along the same column are not significantly different (p<0.05)

Table 2:	Effect of administration of whey on the haematological parameters of healthy albino rats
Control: Rats without whey. PCV: Packed cell volume; WBC: White blood cell. Values are mean of four replicates±SD. Values followed by similar alphabets along the same column are not significantly different (p<0.05)

The significance increase in the haematological parameters of the rats fed with whey was noticeable when the values of these parameters were compared to those without whey (Table 1). Increase in PCV showed that whey causes an improvement in the volume of blood. An increase in PCV might have increase the amount of nutrient the red blood cell can circulate which in turn may be responsible for the increase in the body weight. A high level of PCV is an indication that the rats were not anaemic while a lower level indicates anaemic condition (Schalm et al., 1975). There was also an increase in the WBC. White blood cells especially the lymphocytes are of great importance in defending the body against infection (Schalm et al., 1975). The lymphocytes in the groups fed with 2.5, 2.0, 1.5 and 1.0 mL^-1 of fermented whey significantly increased from 42.5, 40.75, 49.75, 50.0 to 57.50, 57.75, 56.755 and 56.00%, respectively, after been fed with whey The result of the lymphocyte count of the control experiment showed no significant difference.

The safety of whey for human consumption were verified using histopathological analysis and serum biochemical markers such as AST, ALT, ALP, total protein and globulin. The toxicological assessments results in Table 3 revealed that there were no significant increases in the level of all the biomakers such as those obtained for the control experiment. An increase in the blood AST, ALT and ALP has been reported as an indication to both liver damage and possible damage to the heart (American Liver Foundation, 1999; Johnston, 1999; Oyetayo and Osho, 2004; Agarry and Osho, 2005). Meanwhile, from the result obtained, it could be established that whey has no toxic effect on the liver and the heart since no increase was observed in the values obtained. Kelly (1997) also reported that there are no side effects in whey protein with proper supplementation.

In affected liver, both the ALT and ALP levels will be increase (Strove, 1989). The ALP values indicate to the biliary system, either within the liver or in the larger bile channels outside the liver. The ALP is elevated in a large number of disorders that affect the drainage of bile, such as gallstone or tumor blocking the common bile duct. It serves as an indicator of liver damage when there is cholestasis or lack of bile flow (Johnston, 1999). The AST and ALT are enzymes that are located in the liver cells and leak out and make their way into the general circulation when the liver cells are injured (American Liver Foundation, 1999; Johnston, 1999).

Table 3:	Effect of the administration of whey on the biochemical indices of albino rat serum
Group A: Group of four rats orogastrically fed with 2.0 mL of whey, Group B: Group of four rats orogastrically fed with 2.5 mL of whey, Control: Group of four rat without whey. AST: Aspartate-aminotranferase, ALT: Alanine-aminotranferase, ALP: Alkaline-phosphatase. Values are mean of four replicates±standard deviation. Values followed by similar lette(s) along the same column are not significantly different (p<0.05)

Table 4:	Effect of the administration of whey on the albino rat serum protein
Group A: Group of four rats orogastrically fed with 2.0 mL of whey. Group B: Group of four rats orogastrically fed with 2.5 mL of whey. Control: Group of four rat without whey. Values are mean of four replicates±standard deviation. Values followed by similar alphabets along the same column are not significantly different (p<0.05)

The ALP is regarded to be a more specific indicator of liver inflammation, since AST may be elevated in diseases of other organs such as heart or muscle disease. In chronic hepatitis or cirrhosis, the elevation of these enzymes may be minimal and less than 2-3 times normal or moderate 100-300 μ L^-1. Mild or moderate elevations of ALT and AST are non-specific and may be cause by a wide range of liver diseases. AST is widely distributed in body tissues; hence many other diseases involving cellular injury may be accompanied by increases in AST activity for example, severe bacterial infection, malaria, pneumonia, pulmonary infections and tumor (Chessbrough, 2004).

Albumin is produced mainly in the liver and its estimated is a test of liver function (Johnston, 1999). The albumin levels of rats fed with whey were not significantly different (p<0.05) from those of the control. This showed that whey did not increase the biosynthesis of albumin from the liver (Table 4).

To date, no severe adverse reactions have been noted following administration of whey although many individuals sensitive to dairy find that casein is the culprit and they can tolerate whey. Generally, the result of the toxicological assessment revealed that whey contains no toxic substances that may be hazardous to health and its administration improves the health of infected animals.