Evaluation of the Ameliorative Role of Citrullus Lanatus and Annona Muricata Fruit Extracts on Cyhalothrin-Induced Toxicity in Rats

Abstract

The aim of this study was to evaluate the therapeutic role of Fruit Extracts on Cyhalothrin- induced Toxicity using male Sprague-Dawley rats in a completely randomized experimental design. The study was carried out in an experiment that lasted for 35 days. The body weight of the animals was monitored and recorded twice a week throughout the duration of the experiment. Blood samples were collected by ocular puncture into sterile tubes and serum was separated by centrifugation at 2500g for 10mins and stored for determination of some liver biomarkers including ALT, ALP, AST, Total Cholesterol and Glucose using their respective kits. For histopathological analysis of the Liver, The levels of ALP, AST, and ALT significantly p<0.05) increased in groups exposed to Cyhalothrin only but reduced with administration of 100% Citrullus lanatus and Annona muricata extracts. Plate 1A shows the normal architecture of seminiferous tubules and spermatogenic elements in the control group, the Seminiferous tubules and epithelium of the group exposed to cyhalothrin only are characterized by large vacuoles devoid of spermatogenic elements. Testicular tissues of animals co-administered Citrullus lanatus and Annona muricata extracts showed regenerating epithelium filled with maturing spermatozoa. Photomicrograph of the liver epithelium in the control group showed normal architecture of liver cells filled with normal hepatocytes. Cyhalothrin induced oxidative and reproductive stress in exposed animals while from the groups co-administered Citrullus lanatus and Annona muricata the therapeutic role of this indigenous fruit extracts on Cyhalothin-induced toxicity was observed both in the liver and testicular epithelium and therefore can be used as Supplement and suitable first aid for pesticide-related poisoning.

CHAPTER 1

INTRODUCTION

1.1 Background to the study

Insecticides have been used all over the world as a major means of the eradication of insect pests in agriculture, homes, offices, schools, hospitals and other pest-infested areas in developed and underdeveloped countries of the world. Man and other non-target organisms become exposed to the toxic effects emanating from the release of these chemicals into the environment as well as water bodies.  The most widely used insecticides are pyrethroids.

Pyrethroids are a class of synthetic insecticides that mimic the structure and insecticidal properties of the naturally occurring insecticide pyrethrum which comes from the flowers of Chrysanthemums,

Research has shown that 26 to 40% of the world’s potential crop production is lost annually because of diseases, weeds and pests (OECD-FAO, 2012). The use of pesticides has helped to curb this menace. Due to the advantages of the use of these pesticides, developed and developing countries including Nigeria have found it convenient to use them to eradicate insects that cause a nuisance in homes, hospitals and offices resulting in the unregulated and indiscriminate use of these chemicals. All pyrethroids are hazardous but some have higher toxicity levels than others and play a vital role in many degenerative diseases, as well as, impairment of reproductive functions of mammals

A mixture or cocktail of these pyrethroids has been found in agricultural produce  Most of the insecticides such as Deltamethrin  Cypermethrin have been reported to cause reproductive dysfunction, impairment of spermatogenesis, increased sex cell abnormalities and decrease enzymatic antioxidant levels. Lambda-cyhalothrin has LD₅₀ ranging from 79-56mg/kg/body weight in male rats and decomposes in extreme heat (257^0C)

Cyhalothrin is used to control a wide range of pests via diverse methods of application. However, the mechanism of action associated with its use as an insecticide is, primarily the disruption of the nervous system. This could lead to death or paralysis and also pose a threat to the natural environment as well as non-target organisms. It also impairs the development of early life stages of aquatic life, especially fish. Studies have shown that lambda-cyhalothrin causes oxidative stress on the myocardial hyalinization, lysis of reticular framework in the spleen and spermatogonial cell degeneration in testes   Lambda-cyhalothrin is said to be slightly toxic to birds, and highly toxic to fish, invertebrates and bees according to the regulatory bodies. The most vulnerable target organs of this pyrethroid are the gonads, the liver and the nervous system

Watermelon Citrullus lanatus has been reported to be a good source of amino acid citrulline and the antioxidant lycopene, a great source of vitamin A, vitamin C, vitamin B6 and vitamin B.

According to Olagunju and  Annona, muricata possesses high antioxidant activity which qualifies it as a functional drink. They reported that its raw fruit contains 81 % water, 17% carbohydrate, 1% protein and negligible fat and that in 100 grams reference amount, the raw fruit supplies 66% calories, and contains only vitamin C at a significant amount of 25%.

It is reported that high concentrations of Boron, Iron, and Zinc and low concentrations of Manganese, Copper, Lead, and Cadmium are in soursop juice. The soursop fruit extract is reported to have mitigated caffeine-induced reduction  in testicular and epididymal weight, sperm motility, sperm count and increased sperm head abnormality in a mammalian animal  model

1.2 JUSTIFICATION

According to the National Pesticide Information Centre, pyrethroids, including Lambda-Cyhalothrin, disrupt the normal production of signalling hormones, especially of the reproductive system. There is a high prevalence, and use of Lambda-Cyhalothrin as an insecticide in agriculture to protect stored products, in Green houses, on ornamental plants, and on lawns nationwide. In spite of its unregulated and indiscriminate use, there is a paucity of information on the effect of Lambda Cyhalothrin on spermatogenesis and possible oxidative stress on mammals. This work was, thus conceived to assess the potential toxicity of Lambda Cyhalothrin on the reproductive potential and oxidative stress using mature male rats as a model organism and to evaluate the mitigating effect of Citrullus lanatus and Annona muricate extracts as potential antioxidants against its toxicity.

1.3 Aim

Evaluation of the Therapeutic Role of Citrullus lanatus and Annona muricata Fruit Extracts on Cyhalothrin-induced Toxicity.

1.4 Objectives

The objectives of this study are

1. To determine the possible systemic toxicity of Lambda-Cyhalothrin by assessment of the body weight and the weights of vital organ
2. To evaluate the impact of Lambda-Cyhalothrin on endogenous liver antioxidants through the liver biomarker.
3. To evaluate potential Liver injury by Histopathological analysis of the liver and liver injury biomarkers
4. To assess the potential toxicity of the kidney via  its function
5. To evaluate the impact of Annona muricata and Citrullus lanatus as potential antioxidants against systemic toxicity.

CHAPTER TWO

2.0 LITERATURE REVIEW

Looking at all the benefits derived from the use of insecticides in eradicating the menace caused by insects in the agricultural sector, health sector and the entire world economy recorded by the regulatory bodies over the years. However, their possible adverse effect on humans, animals and the entire environment due to persistent increases in the use of these chemicals can never be overemphasised. Lamda-Cyhalothrin has been classified as an endocrine disruptor. In addition, it affects the nervous system of target organisms and contaminates the soil, air and Pyrethroids induce a mammary tumour, and thyroid and also exhibit some mutagenicity in human lung cell culture and genotoxicity  They are reported to induce renal toxicity, hepatotoxicity, reproductive toxicity and oxidative stress in mammals More so, WHO estimated that 20% of the global suicides were through self-poisoning with insecticides. A range of human health hazards such as itching, and skin and eye irritation, are associated with acute exposure to insecticides. Chronic and sub-chronic exposures caused reproductive dysfunction systemic poisoning, endocrine disruption and fatigue

According to previous works of researchers, these pyrethroids induce injuries on the liver, kidney, and reproductive system while enzymatic endogenous biomarkers of the liver and reproductive organs were significantly altered by these chemicals Other studies reported serious detrimental effects on chronic exposure of deltamethrin on vital organs on daily exposure. Though, it may be considered to have relatively low toxicity when compared to organophosphate chemicals Meanwhile, when products that contain high concentrations of pyrethroids are ingested, they cause toxic effects on the organisms. Deltamethrin exposure has been reported to cause a reduction in the weight of reproductive organs, sperm count, sperm motility, and serum Morphometric and structural changes in female genital organs were reported in cypermethrin) reported reproductive dysfunction in females exposed to fenvalerate. These pyrethroids induced vesicular atrophy of the endometrium, inhibit ovulation, and decreased the number of follicular cells, causing atresia of follicles and corpora lutea (Wolansky, et al., 2008).

Synthetic pyrethroids are manufactured in different forms including emulsifiable concentrates, smoke coils, Wettable and Clustable powders, aerosol sprays, Electric mats and others.

Exposure to pyrethroids has been implicated in the inhibition of the proliferation of peripheral blood leukocytes. Some had been shown to possess the potential to impair fertility and general reproductive health, as well as, inhibit spermatogenesis and  sperm production potential

2.1 Effect of Pyrethroids on the Liver

Repeated oral administration of Cypermethrin was found to reduce the level of glutathione (GSH) and the activities of the antioxidant enzymes. While, the level of Aspartate Transferase (AST) and Alanine Transferase (ALT) were increased in the liver extract and plasma of the Cypermethrin–treated animals. The activities of glutathione S-transferase (GST) and superoxide dismutase (SOD) were reported to be significantly decreased due to deltamethrin administration in rats Liver and kidney injury was confirmed by histological changes and liver injury biomarkers

Researchers reported that repeated administration of deltamethrin at a known dose  significantly exhibited harmful effects on the liver and kidney of Wister rats as compared to higher doses

2.2 Effects of Pyrethroids on Reproductive Organs

Reports from other researchers maintained that male rats exposed to  Lambda-cyhalothrin exhibited a significant increase in abnormal morphology of spermatozoa and lipid peroxidation (LPO).

Accordingly administration of lambda-cyhalothrin exerted adverse effects on the development of mice fetuses.  Chronic exposure to cypermethrin disrupted the production of all androgens, increased the percentage of abnormal spermatozoa, and reduced sperm motility, viability and sperm reserves in Sprague-Dawley male rats. According to the researchers’ histopathological evaluation of the testes of cypermethrin-exposed male rats revealed approximately 40% loss of germ cells including interstitial cells of Leydig and Sertoli cells. Dichlorvos has also been reported to inhibit spermatogenesis by interfering in the testicular meiotic division process and causing  spermatogenic cellular degeneration resulting in the loss of  interstitial cells of Leydig  and depopulation of  spermatocytes and  maturing spermatozoa

Citrullus lanatus commonly known as watermelon is a great source of vitamin A, vitamin C, vitamin B6 and vitamin B1, potassium, magnesium, carotenoid antioxidant and lycopene as earlier reported.

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