COMPARATIVE EVALUATION OF THE EFFECTS OF PALM BUNCH ASH AND TRONA ON THE LIVER OF ALBINO RATS

Figure legends

Figure 1 (Group 1; control): Liver section with normal microarchitecture. Its shows distinct liver sinusoids

(marked by green arrows), hepatocytes (marked by blue arrows), blood cells (marked by red arrow) and central vein (marked by black arrow). H&E X400

Figure 2 (Group 2): Liver section showing mild change in tissue architecture. The stained section shows moderate influx of numerous inflammatory cells in the stroma (an evidence of hepatitis; marked by black arrows) and mild cytoplasmic necrosis. The integrity of the central vein appears to be mildly compromised in some areas. H&E X400

Figure 3 (Group 3): Liver section showing moderate signs of necrosis. Some areas appear vacuolated (marked by four headed arrows) with evidence of karyolysis and inflammatory cells (marked by green arrows). H&E X400

Figure 4 (Group 4): Liver section showing dilated central vein (marked by double headed black arrow), less identifiable sinusoids (an evidence of stromal erosion or hepatitis), a few inflammatory cells (marked by blue arrows) and iron deposits (appear as dark-brown deposits), all pointing towards a medical condition known as cholestasis. H&E X400

Figure 5 (Group 5): Liver section showing mild cytoplasmic necrosis, very few inflammatory cells (marked by black arrow) and mild stromal erosion (with few distinct sinusoids). H&E X400

  Table 1: Comparison of Trona and palm burnt ash effects on animal body weight based on group treatments                                                                                                                                    

   Groups                              Day 1                       Day 5                       Day 9                     Day 13                   Day 17                   Day 21                                                                                                                                                    

Overall, table 3 above show insignificant differences in haematological and biochemical parameters within and among the groups (p>0.05), except in lactate dehydrogenase which had significantly increased activity (p<0.05). Significant differences were observed when group 5  was compared with group 1  and group 3  (p<0.05). Insignificant higher levels of Hb, PCV and ALP with corresponding insignificant lower levels of ALT, LDH-P and Amylase activity were observed in group 5  when compared with group 4 (p>0.05), while significant lower levels of AST was also observed in the group 5 (p<0.05). However, insignificant higher levels of Hb, PCV and AST (p>0.05) with significant higher level of LDH activity (p<0.05) and lower insignificant level of ALP, ALT and Amylase activity were observed in group 5 when compared with group 3 (p>0.05). Insignificant increase in Hb, PCV, ALP, AST, LDH-P and serum amylase with insignificant decrease in ALT were observed when group

2 was compared with the control group 3 (p>0.05). Higher insignificant levels of Hb, PCV, LDH-P and serum amylase with corresponding lower insignificant levels of ALP, AST and ALT were observed in group 2 when compared with group 4 (p>0.05). However, lower levels of Hb, PCV, ALP, ALT and higher levels of AST, LDH-P and Amylase were observed in group 2 when compared with group 3 (p>0.05).

Table 4: Correlation analysis between some affected haematological and biochemical parameters

Groups                                                                   Correlation          Remark

  Control variable (Parameters)             r-value      P-value                                       

**Correlation is significant at P<0.01 and P<0.05 (two tailed test), N=30, n=6

Table 4 above shows significant strong positive correlation between Hb and PCV, LDH and AST (p<0.01).

DISCUSSION

The traditional use of food tenderizers (Trona and Palm bunch ash) cannot be abated in local communities, especially in developing countries with low income earners. These food tenderizers are believed to reduce cooking time and thus, reduce expenditure (Nasiru et al., 2011). This study investigated the effect of Trona (Akaun) and Palm bunch ash (Ngu) in relation to body weight, haematological, biochemical and liver microarchitecture of male albino rats. The reduction  in  body  weight  in  the  test  groups when compared with the control group suggests that food tenderizers may cause some health problems. However, the body weights of the test groups administered with Trona were lower than those administered with Palm bunch ash (table 1). The latter suggest that the degree to which the compared food softeners inflict injury to the entire  body  system  varies.  This  reduction  in body weight of the test groups fed with Trona is dose dependent (Table 1) and is in agreement with the study carried out by Ebadan et al., (2014)   who   reported   similar   effect.   The reduction in body weight could be as a result of chemical nature of the substance (trona). More so,  animals  fed  with  Trona  had  lower  liver weight and relative weight (table 2) when compared with the control group which had no pathological   change   in   their   liver   sections (Figure 1). The difference could be adduced to the  mild  toxic  effect  of  trona  on  the  liver, evident in Figure 2. Polycythemia (erythrocytosis) is refers to increased Hb and red cell   count   and   is   associated   with   chronic chemical  exposure,  renal  disease  and dehydration among others (Sood, 2009). This is consistent with the result of this study which revealed dose dependent higher haemoglobin (Hb) and packed cell volume (PCV) estimates in rats fed with Trona when compared with those fed with palm bunch ash and the control group. This suggests that trona may have the potency of dehydrating the body, hence the higher Hb and PCV and the strong positive correlation between them (tables 3 and 4). This is in consonance with the findings of Oyeleke et al., (1981) who observed that high potash intake result in reduction of food and water consumption. This dehydrating effect of trona may be linked to its chemical components which include:   calcite,   hanksite,   halite,   pirssonite, borax, sodium sesquicarbonate, sodium carbonate, sodium sulphate (Copenhafer, 2000; Lu et al., 2005; Ajiboye et al., 2013). The enzymatic activity of ALT, AST, ALP and LDH are  usually  raised  in  acute  hepatotoxicity  but tend to decrease with prolonged intoxication due to  damage  to  the  liver  (Obi  et  al.,  2004; Mcmillin and Johnson-Davis, 2010).   Higher ALP activity was observed in the test groups fed with Palm bunch ash when compared with those fed with Trona and the control group. This correlates well with the studies carried out by Omajili et al., (2010) and Ajiboye et al., (2013) who reported increase in serum levels of ALP following the administration of Trona. This may be suggestive of liver damage as figure 4 and 5 show evidence of hepatitis.   This is further supported by the higher liver weight and organ versus body weight ratio observed in animals fed with PBA. Since AST and LDH have also been reported to be elevated in disease condition such as pulmonary infarction, acute renal infarction and progressive muscular dystrophy (Sood, 2009), the higher serum AST and LDH observed in the groups fed with Trona when compared with the other groups suggests that Trona may also have extrahepatic effect. The strong positive correlation between AST and LDH (Table 4) further lends support to the latter suggestion. The reports of Bankole et al., (2015) and Ebadan et al., (2014) who observed renal toxicity following the administration of trona (potash) further buttresses the argument. More so,   the   elevated   level   of   serum  AST   and increased activity of LDH may be adduced to the cytotoxic effect of trona on the heart muscle as earlier reported by Ochei et al., (2014). Elevated levels of amylase are found in acute pancreatitis  in  first  24  hours  (Kumar  et  al.,

2010) while decreased levels are found hepatitis (Sood, 2009). With acute pancreatitis and hepatitis occurring simultaneously in a system it could be difficult to come up with a diagnostic value for amylase, as one disease condition counters the effect of the other. This might be the explanation for the insignificant differences observed in the test groups fed with trona when compared  with  the  control  group  (Table  3). Since the serum levels of amylase in the test groups  fed  with  PBA are relatively the  same (Table 3), it could be argued that PBA had little or no effect on the pancreatic function at the administered  dosages.  The  iron  deposits observed in figure 4 may be linked to the decreased Hb and PCV observed in Group 4. The  decrease  in  these  haematological parameters may be due to cholestasis caused by intrahepatic haemolysis. Cholestasis has been reported to result in nutritional deficiencies of the fat-soluble vitamins A, D, or K and characterized by elevated serum alkaline phosphatase (Kumar et al., 2010). The latter report is consistent with the results of this study. Surprisingly, lower haematological values were not observed in group 5. This, yet unclear, suggests that the effect of PBA may be dose and duration dependent. In general, considering the microarchitectural changes observed in the liver sections of the test groups when compared with the control group (Figures 1), it is agreeable that PBA (Figures 4 and 5) had as much hepatotoxic effect as trona (Figures 2 and 3) but the observe differences in the biochemical and haematological effect suggests that trona was more toxic.

Conclusion

Based   on   the   body   weight,   biochemical,

haematological and histological evaluations, this study suggests that both Trona and Palm bunch ash are more toxic at 972 mg/kg body weight. Minor  architectural  changes  were  observed  in the experimental animals given both tenderizing agents at 486 mg/kg body weight, thus lower concentrations should be used when preparing meals.

Acknowledgements

Special  thanks  are  to  the  staff  of  Medical Laboratory Services, Madonna University, Elele campus,  Nigeria  and  staff  of  Histopathology Unit, Federal Medical Centre, Owerri, Imo State for allowing us make use of their facility and for their technical support throughout the study.