PRELIMINARY REPORT ON THE ABSENCE OF TOXICITY OF FRUIT PULP OF PALMYRAH (BORASSUS FLABELLIFER L.) IN ICR MICE

Palmyrah (Borassus flabellifer L.) flour prepared from the seed shoot has been previously reported to cause hepatotoxic, neurotoxic and other toxic effects. Although palmyrah fruit pulp (PFP) is also consumed in Sri Lanka, no toxicity studies have been done. Short and long term toxicity studies were therefore conducted with PFP by administration of 10% or 50% PFP (dry basis) incorporated into WHO standard feed (part of the maize being substituted with PFP) compared with a control group of ICR mice, fed the WHO standard feed (n=7 in each group, age 6 weeks, weight 3644g) , for 7 or 30 days. In both studies the effect of 10% or 50% P F P feed on l iver and kidney function and haematological parameters (haemoglobin concentration, packed cell volume, white blood cell count and red blood cell count) were determined. Liver function was assessed by estimation of serum levels of alanine transaminase, aspartate transaminase and alkaline phosphatase. Renal toxicity was assessed by estimation of serum creatinine levels. In the long term toxicity study, effect of PFP on histology of main body organs (heart, lung, liver, kidney and intestine) were also assessed by microscopic examination of haematoxylin / eosin stained sections of these organs. Results of both the short term and long term toxicity studies demonstrated that PFP at the 10% or 50% level did not have any significant effect on kidney or liver function nor on the haematological parameters under study and on the histology of major body organs in ICR mice. In all studies, neurotoxicity was assessed by comparing the behaviour ofthe test group with the control group. No behavioural changes or spasms were observed in the test groups of mice fed with the feed containing 10% or 50% PFP. The overall results suggest that PFP even a t the 50% level in the standard feed probably does not produce any toxic effects in ICR mice.


INTRODUCTION
Palnlyrah (Borassus flabellifer L. , Family; Arecaceae, Sub family; Boracidae) fruit pulp (PFP) contains several bioactive flabelliferins.ls2Flabelliferins are a family of steroidal saponins complexed with carbohydrate m o i e t i e ~. ~ Of the flabelliferins isolated so f a r from P F P , t h e Palmyrah flour (odiyal) prepared from the seed shoot shows n e u r o t ~x i c , ~~ he pa to to xi^,^ i m m u n o t ~x i c ~-' ~ and other biological activities such as ~nutagenic,'~ and clastogenic14 effects in mice, when 50% of the normal feed has been substituted with palmyrah flour.Furthermore, feeding of diets containing palmyrah flour tolactating rats resulted in hepatic, pulmonary and renal lesions and subcutaneous haemorhages in the suckling rats.15Although PFP is not consumed widely as a sweet meat at present, the Palmyrah Development Board (PDB), Sri Lanka intends popularizing other products from P F P (eg: beverages) which will increase its consumption.16No control studies have been carried out to date to investigate possible toxic effects that PFP consumption may produce.Therefore, the study of the potential toxic effects of PFP becomes important.

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The objective of this study was to test for toxic effects of PFP in ICR mice, when substituted in standard feed as follows.

Short term toxicity studies
At 10% feed (7 days) As evident from Table 1, there was no evidence of toxicity as judged by serum enzymology (alanine transaminase, aspartate transaminase, alkaline phosphatase), serum creatinine a n d haematological parameters.No neurotoxic symptoms such as hyper-excitability, salivation, muscle tremors, piloerection, changes in locomotor activity, changes in posture, ataxia and loss of reflexes could also be observed.

Long term toxicity studies
At 10% or 50% feed (30 days) haematological parameters were elevated in the animals receiving feed containing PFP a t 10% or 50% level for 30 days when compared with the control animals.No neurotoxic symptoms could also be observed in the test animals.No histopathological changes were observed in  Results of the present investigation demonstrate that short term (7days) or long term (30 days) Results given in table 2 show that neither the feeding of p ~p incorporated feed to did not serum enzyme levels and creatinine nor the produce any significant toxic effects as assessed by its effects on liver and kidney function, 2. haematologicl parameters and histology of major body organs.This is in contrast to what has been observed in a previous investigation with palmyrah In this study administration of palmyrah flour containing feed (at a 50% level) to mice resulted in neurotoxic symptoms in 4-6 days.This was not observed when mice were fed on 50 3. % PFP containing feed even for 30 days.Palmyrah fruit pulp therefore appears to be non-toxic a t least up to a 50% level of substitution.
Food intake and weight gain of the test animals were normal over a period of one month 4. a t a 10% level and 50% level further confirming the probable absence of toxic principals in PFP similar to those reported in palmyrah flour.

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haematoxylin and eosin stained sections of the liver, kidney, lung, heart and intestine of test animals when compared with those from control animals.Since the predominant toxicity of the flour is on the liver 6photomicrograph sections of mouse livers in control and test animals fed PFP containing feed (at 10% or 50% level) are shown in Figures 1& 2.

METHODS AND MATERIALS mice, while the triglycoside Fb has been shown to
West of Sri Lanka.The fruit pulp was extracted creatinine20 were determined by standard manually as previously de~cribed.~colorimetricmethodsusing commercially available diagnostic kits (Randox Laboratory, Animal Model: Inbred genetically homogeneous, UK).Institute of Cancer Research (ICR) mice ( age 6-8 weeks, weight 35-428) were obtained from the Haematological parameters: At the end of the MRI (Medical Research Institute), Colombo and short term (7 days) and long term (30 days) of experiments were conducted a t the animal house, feeding with the PFP containing feed, the animals University of Sri Jayewardenepura.wereanesthetised with diethyl ether and the blood samples (lml) collected into clean dry sample Male ICR mice were used for each variable bottles containing anticoagulant EDTA, by cardiac (n=7 in each group).The control animals were puncture.The haematological parameters (packed fed with the standard rat and mouse breeding cell volume, red blood cell count, white blood cell feed prepared by the Medical Research Institute, count and haemoglobin concentration) in the blood according to WHO rat and mouse breeding feed samples were assessed by the methods described formula.17Thetest animals were fed with the 10% by International Committee for Standarization in or 50% PFP incorporated in the above WHO haematology21 and compared with corresponding standard feed.The 10% test feed was prepared values in control animals.byreplacing a part (one third) of the maize in the WHO standard feed with PFP ( on a dry weight Hist~pathology~~: The animals were sacrificed and basis).The test and control diets were isocaloric.theliver, kidney, heart, lung and intestine were Palmyrah fruit pulp: PFP was obtained from possess anti-microbial activity.'fruits originating from Kalpitiya in the North-