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Haematological and Biochemical Changes in Chicks Fed with Aspergillus terreus Infested Feed

Influence of Aspergillus terreus infested feed on the growth and metabolism of chick was investigated. The relative weight gain of different organs and feed intake gradually decreased with the increased amount of intake of infested feed. Serum Glutamate Pyruvate Transaminase (SGPT) and Serum Glutamate Oxaloacetate Transaminase (SGOT) activities and bilirubins increased with increase of intake of A. terreus infested feed, indirectly with territrem B intake. However, proteins, iron, urea, calcium and albumins with increased exposure to A. terreus infested feed. Different biochemical components of blood also decreased with increased A. terreus infested feed intake by the chicks.


INTRODUCTION
Natural contamination of foods and feeds with mycotoxins is widespread and thus the mycotoxins reach human beings through a food chain. The ingested mycotoxins affect the health of man varying from acute to chronic. Fung and Clark (2004) elegantly discussed various aspects of health hazards of man caused by mycotoxins. The inhalation of dust containing mycotoxins can also cause a variety of toxic effects humans (Bunger et al., 2004). Mycotoxins also cause severe damage to the vital organs. Mycotoxins such as aflatoxins (hepatotoxic, carcinogenic), zearalenone (estrogenic and teratogenic), ochratoxin A and citrinin (nephrotoxic), DON (feed refusal and vomiting), penicillic acid and citroviridin (cardiotoxic), T2 toxin (mouth lessions and loss of appetite), fumonisins (neurological disorders and liver damage), penitrems and territrems (tremorgenic) and terreic acid (diabetogenic) are reported to cause a wide range of health hazards (Ratcliff, 2002). Some mycotoxins are also reported to cause genetic disorders in animals and plants (Fink-Gremmels and Malekinejad, 2007). Galvano et al. (2001) reported that fumonisins cause DNA damage in human fibroblasts. Morgavi and Riley (2007) have excellently reviewed the role of fusarial toxins in animal health.
Similarly, a large number of workers including (Chowdhury et al., 2005;Akande et al., 2006;Abbes et al., 2006;Sharma et al., 2008;Che et al., 2011;Iheshiulor et al., 2011) have investigated biological effects of different mycotoxins in chicks. Reddy et al. (1997) have reviewed the incidence and biological effects of tremorgenic mycotoxins on animals. Rafiyuddin et al. (2006) have reported the toxicity of tremorgenic mycotoxins in chicks. However, only limited information is Asian J. Poult. Sci., 9 (3): 172-178, 2015  Weeks Treated Control available with regard to toxicity of territrem B, produced by A. terreus on humans and animals (Jiang et al., 2005). Therefore, in the present investigations an effort was made to investigate the effect of territrem B on growth and development of chicks.

MATERIALS AND METHODS
Six week old chicks with their pre recorded initial average weight were selected and randomly assigned to five groups. They were fed with Aspergillus terreus infested feed for 6 weeks. The infested nature of feed was tested for the presence of A. terreus and territrem B. At the end of 2, 4 and 6th weeks of incubation, a set of chicks were sacrificed and analyzed for selected parameters. Blood was collected for haematological and biochemical analysis. Standard methods were adopted for analyzing blood serum amylase (Gomori, 1957), serum proteins (Kingsley, 1942), serum bilirubin (Jendrassik and Grafs, 1965), serum calcium (Clark and Collip, 1925), serum iron (Otto, 1958), serum urea (Wybenga et al., 1971), SGPT and SGOT (Reitman and Frankel, 1957), blood iron and haemoglobin (Wong, 1928) and blood sugars (Folin and Wu, 1920). The RBC and WBC counts were made with the help of haemocytometer. Biochemical changes such as glycogen (Klicpera et al., 1952), tissue proteins (Lowry et al., 1951) and total cholesterol in serum and tissue (Zlatkis et al., 1953) were also determined and body weight of chicks was also recorded.

Statistical analysis:
The experiments were conducted thrice each with three replicates. The results obtained were subjected to statistical analysis using SPSS package (12.0 version) at p = 0.05 level of significance. The p-value less than 0.05 (p<0.05) were taken as significant.

RESULTS AND DISCUSSION
The results pertaining to feed intake/wastage are presented in Fig. 1 and other results are presented in Table 1, 2 and 3. A critical perusal of Fig. 2 reveals that weight of heart and liver decreased due to intake of A. terreus infested feed, which however, showed slow recovery with the progress of age. Weight of the brain increased after 2nd week of feed intake. On the other hand, weight of kidney increased marginally with the age of the chick (p<0.05). Increased organ weight observed in this study has also been recorded by other investigators (Chowdhury et al., 2005). Reports are also available on increased gizzard and bursa weights in broiler chicks fed with Fusarial mycotoxins like DON and T 2 toxins (Huff et al., 1986;Kubena et al., 1989). Aflatoxins were also reported to cause increase in liver, spleen and kidney weight of broiler chicks (Huff et al., 1986(Huff et al., , 1988. Norred and Voss (1994) and Henry et al. (2000) have recorded more intense symptoms        such as diarrhea, decreased feed consumption, decreased body weight and increased relative weights of liver in broilers. Girish et al. (2008) also reported the decreased weight of turkey poultry fed with fusarial mycotoxins. Changes in the components of blood serum are presented in Table 1. Significant changes in the composition of blood serum were recorded due to intake of A. terreus infested feed. However, the degree of changes varied with the amount of feed intake. An increase in the amylase activity was recorded with the progress of growth of chick and feed intake. On the other hand, serum proteins decreased under the influence of A. terreus infested feed. Oguz et al. (2002), Allameh et al. (2005) and Safameher (2008) have also reported decreased serum protein in chicks fed with aflatoxin contaminated corn. A positive correlation could be observed with the intake of A. terreus infested feed and protein content of serum. Iron, urea, calcium and albumin levels decreased with the increase of intake of infested feed. A significant increase in serum bilirubins, SGPT and SGOT was recorded with the increased feed intake (p<0.05).
The effect of A. terreus infested feed on biochemical and haematology composition of blood was assessed and the results are presented in Table 2. The results clearly reveal a significant decrease in blood iron, haemoglobin and sugar. The decrease was progressive with the increase of feed intake. Reddy et al. (1997) have also recorded decreased haemoglobin, albumins, proteins and sugar of the blood of chicks under the influence of gliotoxin. The RBC, WBC, neutrophil and lymphocytes decreased with the increased exposure of A. terreus infested feed (p<0.05). Monocytes were affected only marginally. However, basophils number increased by the end of 6 weeks observation. The present observations are in agreement with those of Singh et al. (1992), who also recorded decrease in WBC counts of chicks receiving aflatoxin B 1 . Verma et al. (1991) reported significant changes in the blood of rat under the influence of aflatoxins. In 1995, a number of histopathological changes in chick receiving trichothecene mycotoxins were reported. Reddy et al. (1997) have also recorded an increase in Serum Glutamate Pyruvate Transaminase (SGPT) and Serum Glutamate Oxaloacetate Transaminase (SGOT) of blood of chick under the influence of gliotoxin produced by Trichoderma viride. Asian J. Poult. Sci., 9 (3): 172-178, 2015 Biochemical changes in different vital organs of chick under the influence of A. terreus infested feed were analyzed and the results are presented in Table 3. The glycogen content of liver decreased with the increased concentration and exposure time of territrem B, glycogen of kidney, brain and heart decreased marginally under the influence of territrem B. Protein content of liver, kidney brain and heart also decreased with the increased intake of A. terreus infested feed (p<0.05). On the other hand, cholesterol content in liver increased with the intake of A. terreus infested feed as recorded by Jassar and Singh (1993) have also recorded significant biochemical changes in the blood of broiler chicks treated with aflatoxins.

CONCLUSION
Feeding of chicks with Aspergillus terreus infested feed has significant influence on the vital organs in particular and overall growth of chick in general. Feed efficiency decreased with the age of chicks. Significant changes in the composition of blood serum were noticed. Serum bilirubins, SGPT and SGOT increased with the progress of growth of chick till the end of observation period. Similarly iron, haemoglobin and sugar contents in the blood decreased. Similarly, RBC, WBC, neutrophils and lymphocyte count also decreased with the increased intake of infested feed. However, basophils number increased. Significant decrease in glycogen, protein contents of the liver, kidney and heart decreased. On the other hand, cholesterol contents of liver increased. These observations confirms the toxicity of territrem B produced by A. terreus in the infested feed.