Assessment of the haematological parameters and biochemical indices of albino rats fed with spores of Trichoderma viride

. The haematological as well as some enzymes biomarkers of albino rats orogastically dosed with different concentrations of Trichoderma viride as well as T. viride subjected to various treatments were evaluated. Generally, red blood cell count, haemoglobin content and packed cell volume of the control group were significantly higher than that of animals dosed with different concentrations of T. viride. The same trend was observed when T. viride was amended with 1% lactose, sodium nitrate and sodium chloride. There were no significant differences in the alanine aspartate, alkaline phosphatase and alanine transaminase values of animals dosed with lower concentrations of T. viride. This study suggests that lower concentration of T. viride could be used in controlling fungal phytopathogens.


Introduction
pathogen for nutrients and space or induced plant resistance (Daayf et al., 2003).
It employs natural enemies of pests or pathogens to eradicate or control their population. The induction of plant resistance using non-pathogenic or incompatible microorganisms is also a form of biological control (Cook, 1993;Schouten et al., 2004).
Trichoderma spp. are free-living fungi that are common in as well as on plant surfaces (Islam et al., 2011). Trichoderma species lytic enzymes such as chitinases, glucanases and proteases which are essential in biocontrol of plant diseases (Gajera and Vakharia, 2012). Previous study by Ekundayo et al. (2015) has shown the antagonistic activities of T. viride against Sclerotium rolfsii, soil borne pathogen of many agricultural and horticultural crops. However, there is dearth of information on the toxicological effects of this particular fungus. Therefore, this present investigation was conducted to determine the effects of this fungus on the haematology and some enzymes biomarkers of albino rats.

Fungal isolates and treatment conditions
Trichoderma viride, an isolate of maize cob was obtained from the Department of Microbiology, The Federal University of Technology, Akure (FUTA). The fungus was cultivated using LAB M potato dextrose broth in conical flask while other flasks containing T. viride was amended separately with 1% NaNO3, lactose and NaCl.

Experimental animals
A total number of 30 albino rats were obtained from the Department of Animal Production, The Federal University of Technology, Akure, and were caged in a well ventilated conditions. Prior to the experiments, the animals were weighed and stabilized for a period of 8 days by giving them grower mash prepared by Guinea Feed Nigeria Limited and water ad libitum.

Animal grouping and administration of T. viride
The animals were grouped into ten with three rats in a group for a period of 7 days. The treated albino rats were orogastically dosed with the different treatments: 1, 0.5, 0.1 mL T. viride, 1 mL T. viride supplemented with 1% NaNO3, 1 mL T. viride supplemented with 1% NaCl, and 1 mL T. viride supplemented with 1% lactose. There were 4 sets of control, (1) animals fed with 1 mL NaNO3, (2) those fed with 1 mL lactose, (3) those fed with 1 mL NaCl, and (4) those fed with clean water only. After the expiration of 7 days of treatment, the animals were sacrificed and their haematological parameters and enzyme biomarkers were determined (Adebolu et al., 2011).

Determination of liver functioning tests
Some liver functioning tests, aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP) were determined according to the protocols of Babson et al. (1966) and Momoh et al. (2013).

Data analysis of results
The data collected were then subjected to Analysis of variance (ANOVA) and the means were separated by Duncan's New Range Multiple Test at P = 0.05, using SPSS version 15.0 Microsoft Windows XP. Table 1 shows the haematological parameters of albino rats subjected to various treatments. The results show a significant difference in the erythrocyte sedimentation rate of all the treatments. The highest erythrocyte value was obtained from NaNO3+ T. viride. There was no significant difference in the packed cell volume of albino rats fed with basal diet and lactose. The least PCV was obtained from rats fed with NaNO3+ T. viride. Also, the highest haemoglobin content was obtained from control group although there was no significant difference in the haemoglobin content of lactose and NaCl fed albino rats. The white blood cell count of albino rats fed with 1.0 mL of T. viride was significantly higher than that of control group. The white blood cell differential count of albino rats treated with different concentrations of T. viride and T. viride supplemented diets presented in Table 2. Lymphocytes of control and albino rats fed with 0.1 mL of T. viride were significantly higher than other treatments. Also, there were significant differences in the monocytes, neutrophils, eosinophis and basophils of the different treatments.

Biochemical indices of albino rats treated with T. viride
The AST values of animals fed with different concentrations of T. viride and T. viride supplemented with sodium chloride were significantly lower than the control. However, there was no significant difference in T. viride supplemented with lactose and sodium nitrate as well as the control (Figure 1). The ALP and ALT of T. viride supplemented with lactose, sodium chloride and sodium nitrate were higher than the control (Figures 2 and 3).

Discussion
Haematological parameters are often used to determine the negative effects of a foreign substances on the blood. There was significant increase in the red blood cell (RBC), haemoglobin concentration (Hb) and packed cell volume values of the untreated rats (control group) similar to the findings of Fasanu and Oyedapo (2008) and Ogunmefun et al. (2013). The decrease in RBC of treated group suggests that T. viride does not relatively possess haematinic property but causes anaemia. It can also be an indication that T. viride impaired the production of blood cells or the rate of destruction of matured red cells was greater than the rate of its production (Odeyemi et al., 2008).
Neutrophils which are matured cells usually attack and destroy bacteria in the blood (Dacie and Lewis, 1995). The significant increase in the neutrophils implied enhancement in the ability of the blood component to phagocytose. "Lymphocytes are the main effector cells of the immune system" (McKnight et al., 1999). Lymphocyte values were lowered in the treated groups showing that the immunological status of the body was not enhanced.
The biochemical indices are useful parameters in assessing the functionality of the liver and kidney (Odeyemi et al., 2008). The ALP is a marker enzyme of damage for plasma membrane and endoplasmic reticulum (Wright and Plummer, 1974;Shahjahan et al., 2004). Enzymes from diseased of damaged tissues may become recognizable in the serum presumably by leakage through altered cell membrane of the rat organs (Akanji and Ngaha, 1989). Supplementation of T. viride with sodium chloride, lactose and sodium nitrate caused a significant increase in the ALT and ALP indicating liver damage. The increase in ALT and ALP of rats fed with 1.00 mL of T. viride was an indication of the damage inflicted on the plasma membrane. "This increase may be due to distruption of the ordered lipid-bilayer of the membrane structure of the affected organ " (Yakubu et al., 2003). Decrease in alanine aspartate (AST) values of the different concentrations of T. viride proved there was no damage to the organs.

Conclusion
This research has shown that T. viride had dose and parameter specific effects on the haematological and biochemical parameters investigated. I equally acknowledge all authors whose works have been cited in the course of preparation of this manuscript.