|Investigation on Antihepatotoxic Effect of Neem Leaves|
Dr. Ramnath D. Sadekar, Dr. Sunil P Waghmare
Dr. Syed Zakir Ali, Dr. Prafull A. Zade
The Indian livestock industry is primarily dairy based. Governmental, non-governmental and private efforts have brought about a white Revolution in the country making India the largest milk producer in the world. However, the problem of early age weight loss in calves - leading to subsequent low productivity - has been one of the constraints faced by the industry. Among the many causes of early weight loss, sub-acute liver toxicosis has been identified to be the most prevalent. Environmental factors like contaminated water, insecticide residues in feeds as well as fodders and fungal contamination of stored feed are mainly responsible for inducing liver toxicosis. Considering the importance of liver in various physiological processes in the body, correction of liver toxicosis is one of the prime measures for improving early age health of the calves and subsequent productivity of dairy animals in this country.
Though a large number of pharmaceutical products containing so-called liver protectants, mainly of plant origin, are available, their costs are prohibitory and beyond the economic capabilities of the livestock owner. These products also lack scientific validation of the claims that are made regarding their utility. It has therefore become essential to introduce such drugs which are not only scientifically validated, but also easily available and cheap, so that farmers can use them extensively.
The Ayurveda describes Neem as one of the easily available medicinal plants which has liver stimulating and liver protective properties. The importance of Neem in health management has been recognised the world over and extensive research has been going on to evaluate its therapeutic properties in laboratory animals and in human beings. However, the utility of neem leaves in livestock diseases has not been explored so far. The present investigation was undertaken to evaluate the efficacy and utility of this all important tree in correcting the major disease entity in an important group of livestock in this country i.e. young calves.
The overall objectives of this study were - to introduce low grade sub-acute liver toxicity in calves, using carbon tetrachloride. To study the microchanges which occur in liver toxicity in relation to hematological, blood biochemical and liver enzyme profile. To study the efficacy of dry neem leaves in reversing the toxicity in the liver and in restoring the normal liver function in calves.
1) Experimental Animals: Ten cross-bred male calves (1-2yrs), healthy (80-100 Kgs) fed normal diet and dewormed, were divided in two groups.
Group I - Untreated control (T1) - induced toxic with carbon Tetrachloride (0.5 ml / kg) intraruminally and kept untreated
Group II - Neem leaves treated (T2) - induced toxic as above and fed 24 hours after dried neem leaves powder @ 24 gm / day / animal daily for 7 days. The leaves powder was given orally in water.
2) Parameters Studied: (Clinical, lab & liver function)
b) Laboratory investigation
c) Liver Function Test - BSP clearence
3) Statistical Anaysis - FCRD
Administration of CCl4 produced clinical toxicity in all the experimental calves within 24 hours and was indicated by rise in body temperature, pulse rate and respiration. There was general depression, anorexia and partly semi-solid faeces. Dried neem leaves powder was administered 24 hours after the initiation of toxicity and continued for 7 days.
The results were -
1) Introduction of toxicity increased body temperature, pulse rate and respiratory rate significantly in all experimental animals. Administration of neem leaves lowered the raised body temperature to normal within three days of treatment. In untreated controls, fever pulse and respiratory rate returned to normal in 7 days.
2) CC14 induced toxicity lowered Hb and TEC values with increase in PCV significantly. The treatment with neem leaves did not have significant effect in restoring these values.
3) The plasma glucose and serum total protein levels were significantly lowered 24 hours after induction of toxicity. Administration of neem leaves initiated rise in these levels within 48 hours after treatment and pretoxicity levels were attained within a week.
4) Serum bilirubin level was significantly raised in all the calves induced hepatotoxic. However, treatment with neem leaves restored these levels to normal within a treatment period of one week. The Serum bilirubin level remained higher in untreated calves indicating continued toxic effect of carbon tetrachloride.
5) The serum levels of all liver enzymes under study were significantly raised within a period of 24-48 hours indicating liver damage of varied degree. In untreated control group, the enzyme levels in serum remained significantly higher than the pretoxicity levels. Administration of neem leaves powder initiated reduction in levels in respect of SGOT (72 hrs), SGPT (96 hrs), Acid phosphatase (96 hrs), Alkaline Phosphatase (48 hrs), Lactate dehydrogenase (96 hrs) and Sorbitol dehydrogenase (168 hrs), restoring them to normal at the end of the treatment.
6) In all the calves induced toxic, the BSP clearance was significantly reduced and plasma half-life of BSP was significantly raised. Oral administration of dried neem leaves powder restored the BSP clearance capacity of the liver to normal
Discussion: Azadirachta indica has been universally accepted as a wonder tree having use as a healer in many disease conditions of skin, metabolism, gastrointestinal system, cardiovascular system and nervous system. The most valuable pharmacotherapeutic property of this plant has been its liver protecting / liver stimulating effects, which have been studied in laboratory animals and even in human beings in hepatic disease conditions like viral hepatitis and cholelithiasis. However, therapeutic usefulness of neem has still to make a headway in livestock diseases. The present investigation indicates usefulness of neem in its crude form (dried neem leaves) in restoring to normal the disturbed liver function in young calves.
Carbon tetrachloride has been used in this investigation as a classical hepatotoxicant to produce liver damage in calves, without producing mortality. The toxicity produced by CC14 presented a classical picture of altered liver function manifested by changes in clinical, hematological, blood biochemical and liver enzymes profile. The induction of toxicity was clinically manifested by inappetance, dullness, reluctance to move and pasty diarrhoea. In untreated controls, the clinical signs continued over a week's period and in neem leaves treated group the clinical improvement was conspicuous within 48 hours of initiation of therapy.
The rise in body temperature, pulse and respiratory rate in toxic animals were due to inflammatory changes in the liver accompanied with pain in the region and also due to blood changes leading to anemia. The treatment with neem leaves seems to have reduced the inflammatory process within 3 days, thereby bringing back body temperature to normal. The blood biochemical profile as it relates to plasma glucose, serum total proteins and serum bilirubin is a measure of liver function and is always considered in estimating the extent of liver damage by toxicant so also for evaluating the efficacy of drugs which reverse the liver damage. The feeding of neem leaves in the present investigation restored the altered values of these biochemicals within a week, indicating anthihepatotoxic effect of neem. Liver is a seat of large number of varied specific and non-specific enzymes and these enzymes act within the confines of the liver parenchyma. Any increase in the presence of liver enzymes in the serum
leakage of enzymes from the liver parenchyma due to hepatic cellular damage or change in the memberance permeability. Estimating serum enzyme activation of SGPT, SGOT, AP, ALP, LDH & SDH is primarily helpful in evaluating the extent of liver damage. Simultaneous multiple serum enzyme study as conducted in the present investigation indicated efficacy of neem leaves in restoring the altered serum profile of liver enzymes within a period of 96-120 hours.
Bromosulphalenin (BSP) is a dye which is used to measure the excretory capacity of liver. The hepatic damage of minutest nature disturbs the ability of the liver to excrete the foreign substance. The neem leaves have exhibited the hepatoprotective effect on the cellular function by restoring the altered BSP clearance mechanism in damaged livers. BSP clearance is the additional marker which has indicated the usefulness of neem leaves in liver protection. The exact mechanism through which neem leaves bring about liver protectant effect is a matter of future research in livestock. Most of the liver-protectants particularly of plant origin are supposed to produce their action by promoting the formation of P-450 and b5, which are microsomal proteins having antioxidant effects in the liver cells. Some of the drugs are known to promote lipoprotein synthesis and synthesis of ribosomal RNA and t RNA. It is a matter of interest to know whether constituents of neem leaves act in the similar manner.
Conclusion: The present investigation has clearly indicated the usefulness of dry powder of neem leaves in restoring the disturbed liver function in young calves. Administration of dry neem leaves powder @ 20 gm / day / calf brings about the desired effect within a treatment period of one week. These findings have a special significance and high utility under Indian conditions where lack of optimum management and environmental pollution constantly expose younger livestock to hazards like liver toxicity and early age weight loss. Neem being a local house-hold tree, the leaves of which are freely available and easy to administer, should find a regular place in livestock therapeutics, particularly of young growing animals.
Chemical Composition of Neem Leaves
Amino Acid content of Neem Leaves
Source:Wealth of India Vol. - 1 : A
CSIR, New Delhi
Pathak and Kamra, 1989