ANTIDIARRHOEAL EFFECT OF UNRIPE MUSA PARADISIACAE PULP AND PEEL HOMOGENATES ON CASTOR OIL-INDUCED DIARRHOEA IN WISTAR ALBINO RATS

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ABSTRACT

Musa paradisiacae  commonly known as plantain is a rhizomatous  perennial  crop used as a source of starchy staple for millions of people in Nigeria. Different parts of
the plant have been used in the treatment of various ailments and there are claims that it has antidiarrhoeal activity. This study is therefore aimed at determining the effects of  unripe  Musa  paradisiacae  pulp  and  peel  homogenates  on  castor  oil-induced diarrhoea in Wistar albino rats. The qualitative phytochemical constituents of Musa paradisiacae   pulp   and   peel  were   found   to   be   flavonoids,   saponins,   soluble carbohydrates,  tannins, reducing sugars,  hydrogen  cyanide, steroids, alkaloids, and glycosides. The LD50 results showed no toxicity up to 5000 mg/kg body weight. Rats were divided into 7 groups of 4  rats each. The groups were pre-treated as follows: group 1: normal saline (control); group 2: 3 mg/kg lomotil (standard drug); groups 3 and 4: 200 and 400 mg/kg unripe Musa paradisiacae pulp homogenates respectively; groups 5 and  6:  200 and 400mg/kg  unripe Musa paradisiacae  peel homogenates respectively;  group  7:  combination  of  unripe  Musa  paradisiacae  pulp  and  peel homogenates  (200/400  mg/kg  respectively).  After  the  treatments,  diarrhoea  was induced using castor oil. Relative to the control group 1, the  treatment groups 2-7 inhibited  castor  oil-induced  frequency  of  defecation  and  wetness  of  stool  dose dependently  but  non-significantly  (p>0.05).  Both  the  pulp  and  peel  homogenates produced non-significant decreases (p>0.05) in the distances travelled by the charcoal meal (marker) in castor oil-induced diarrhoea rats compared to the control group 1. Pre-treatment of the rats with unripe Musa paradisiacae pulp and peel homogenates decreased significantly (p<0.05) enteropooling indicated by decreases in the volume and weight of the gastro-intestinal contents relative to the control group 1. Treatment with the  unripe  Musa  paradisiacae  pulp  and peel homogenates  led to significant decreases (p<0.05) in the bicarbonate ion concentrations except in group 3 rats while the potassium ion concentrations  increased  significantly (p<0.05) in all  the  groups except in groups 3, 4 and 7 rats which showed non-significant  decreases (p>0.05) compared to the control group 1. Sodium ion concentrations of the pre-treated groups increased non-significantly (p>0.05) except in groups 4  and 7 rats which decreased non-significantly (p>0.05) relative to the control group 1.Using everted rat intestines, the pulp and peel homogenates enhanced significant (p<0.05) influx of sodium ions into the everted sacs (serosal) and significant (p<0.05) efflux of potassium ions out of the sacs (mucosal) in relation to the control group 1. These findings reveal that unripe Musa  paradisiacae  pulp  and  peel  exhibit  antidiarrhoeal  properties  by  inhibiting gastro-intestinal motility, enteropooling, wetness and frequency of defecation. They have  also  shown  abilities  to  facilitate  transport  of  electrolytes  across  the  small intestinal membrane.

CHAPTER ONE

INTRODUCTION

The use of traditional  medicines  in West Africa  is probably as old as  the duration of human settlement in the region (Abdul-aguye, 1997). A medicinal plant provides an important source of new chemical substances with potential therapeutic effects.  These have been used in traditional medicine  for  the treatment  of several diseases and aliments (Mukerjee et al., 1998). It is already important to the global economy with demand steadily increasing not only in developing countries but also in industrialized countries (Sofowara, 1993).

Herbalism or herbal medicine is the use of plants for medicinal purposes, and the study of such use (Briskin, 2000). Herbal medicine is still the mainstay of about

75 – 80% of the world population, mainly in the developing countries, for  primary health care (Kamboj, 2000). Plants have been the basis for medical treatments through much of human history, and such traditional medicine is still widely practiced today (Briskin, 2000).  This is primarily because of the general belief that herbal drugs are without any side effects besides being cheap and locally available (Gupta and Raina,

1998). Modern medicine recognizes herbalism as a form of alternative medicine as

the practice of herbalism is not strictly based on evidence gathered using the scientific method (Talalay, 2001).   According to the World Health Organization (WHO), the use of herbal remedies throughout the world exceeds that of the conventional drugs by two to three times (Evans, 1994). The use of plants for  healing purposes predates human history and forms the origin of much modern  medicine. Modern medicine, does, however, make use of many plant-derived compounds as the basis for evidence- tested pharmaceutical  drugs, and  phytotherapy works to apply modern standards of effectiveness  testing to herbs  and medicines  that are derived  from natural sources (Talalay,  2001). Examples  include aspirin (willow bark), digoxin (from foxglove), quinine (from cinchona  bark), and morphine (from the opium poppy) (Vickers and Zollman,  1999).    Currently,  a number of medicinal  plants with antidiarrhoeal  and antimicrobial properties are used in traditional herbal practice in many countries of the world.  So it is important  to identify and evaluate commonly available  natural drugs that could be used against any type of diarrhoeal disease.

A number of herbs are thought to likely have adverse effects (Talay,  2001). Furthermore,  “adulteration,  inappropriate  formulation,  or  lack  of  understanding  of plant  and  drug interactions  have  led  to  adverse  reactions  that  are sometimes  life

threatening  or  lethal  (Elvin-Lewis,  2001).   Proper  double-blind  clinical  trials  are needed  to  determine  the  safety  and  efficacy  of  each  plant  before  they  can  be recommended  for medical use (Vickers, 2007). Although many  consumers believe that  herbal  medicines  are  safe  because  they  are  “natural”,  herbal  medicines  and synthetic drugs may interact, causing toxicity to the patient. Herbal remedies can also be dangerously contaminated, and herbal medicines without established efficacy, may unknowingly be used to replace medicines that do have corroborated efficacy (Ernst,

2007). The  World Health Organization (WHO), the specialized agency of the United Nations (UN) that is concerned  with international  public health,  published  quality control  methods  for  medicinal  plant materials  in 1998  in  order  to  support  WHO Member  States  in  establishing  quality  standards  and   specifications   for  herbal materials,  within  the  overall  context  of  quality  assurance  and  control  of  herbal medicines (WHO, 2010).

There are different methods of herbal preparations and the exact composition of an herbal product is influenced by the method of extraction. They are:

1)   Tisanes or herbal teas; are the resultant liquid of extracting herbs into water (Green, 2000). The methods used are, infusions (hot water extracts of herbs), decoctions (long term boiled extracts usually of harder  substances like roots and bark) and maceration (old infusion of plants with high mucilage content) (Green, 2000).

2)   Tinctures; alcoholic extracts of herbs generally stronger than tisanes (Green,

2000).

3)   Syrups; extracts of herbs made with syrups or honey (Green, 2000).

In developing countries, diarrhoea continues to be one of the leading causes of mortality and morbidity in children less than 5 years old. According to World Health Report,  diarrhoea  is  the  cause  of  3.3%  of  all  deaths.  Worldwide  distribution  of diarrhoea  accounts  for  more  than  5-8  million  deaths  each  year  in  children.  The incidence  of  diarrhoeal  disease  still  remains  high  despite   the  effort  by  many government  and international organizations  to reduce it.  Nigeria, the fourth largest economy in Africa with an estimated per capita income of $350 has over half of its population living in poverty (WHO, 2007).  This implies that very few people can afford orthodox medicine in curing diseases. Use of traditional medicines to combat the consequences of diarrhoea has been emphasized by WHO in its Diarrhoea Control Programme. It is therefore important to identify and evaluate available natural drugs

as alternatives to current antidiarrhoeal drugs, which are not always free from adverse effects. Several studies have shown the beneficial effects of a number of medicinal plants used traditionally  in the treatment  of diarrhoeal  disease,  one of such being Bombax buonopozense (Akudor et al., 2011), Vitex doniana (Ukwuani et al., 2012), Anacardium occidentale (Omoboyowa et al., 2013) etc.

Musa paradisiacae belongs to the Musaceae family and is cultivated in many

tropics and subtropical countries of the world. It ranks third after yams and  cassava for sustainability in Nigeria (Akomolafe and Aborisade, 2007). Musa paradisiacae is a rhizomatous perennial crop used as a source of starchy staple for millions of people in  Nigeria  (Adeniyi  et  al.,  2006).Unripe  Musa  paradisiacae,  which  is  the  green plantain contains more starch than the ripe plantain in which the starch is converted to sugars (glucose, fructose and sucrose). It has been  indicated  to posses antidiabetic (Eleazu  et  al.,  2013),  antioxidant  (Shodehinde   and  Oboh,  2012),  antimicrobial (Hossain et al., 2011), and antiulcerogenic properties (Ralph et al., 1984). There have also been traditional claims that unripe Musa paradisiacae can be used in diarrhoeal treatments even though it has not been scientifically proven.

1.1 Musa paradisiacae

Fig. 1: Musa paradisiacae fruit (Gibert et al., 2009).

1.1.1 Taxonomy of Musa paradisiacae

Kingdom–     Plantae Division –    Spermatophyta Sub-division – Angiospermae Phylum – Tracheophyta

Class – Liliopsida Order – Zingiberales Family – Musceae Genus – Musa

Species – Paradisiacae

(Smith, 1977).

1.1.2 Common names of Musa paradisiacae

Musa  paradisiacae  is  commonly  known  as plantain.  Among  the  Igbos  of Nigeria, it is known as “ogede or abrika”, in Yoruba as “ogede agbagba”, in “Igala as agbo̥” , and in Hausa as “agada or afutu”.

1.1.3 Origin of Musa paradisiacae

Bananas and plantains belong to the genus Musa. It was Linnaeus that  first gave the scientific name Musa sapientum  for all sweet bananas, and the  scientific name Musa paradisiacae for plantains (Simmonds, 1962). However, Linnaeus did not know that the two species he had described were in fact hybrids and not two distinct species (Zeller, 2005). Therefore, those two names could not be relevant in modern taxonomy.

Genetic studies have then demonstrated that all edible bananas and  plantains come from a common ancestor, Musa acuminata. Plantains also carry  genes from another ancestor, Musa balbisiana (Lejju et al., 2005). The genome of each ancestor could be represented respectively by the letter A and B. Then, further studies showed that  edible bananas  are mostly triploids  and their  genome  would be described  as AAA.  This  means  that  they  carry  three  sets  of  chromosomes  derived  from  M. acuminate  (Simmonds,  1962). Different  hybrid  combinations  have been observed, such as AAB, BBB, and tetraploid groups (AAAA) were also described.

Therefore, an accurate classification for bananas seems to be a great challenge. However, one thing sure in that banana taxonomists seem to agree  that there is no single scientific name that can be attributed to all edible bananas (Zeller, 2005; Solofo and Ellis, 2009). Therefore, a new type of classification was proposed by Simmonds and that would abandon the Latin name to use instead a  group indication like this: genus  (Musa)  +  genome  group  (e.g.  AAA)  +  subgroup  name  (e.g.  Cavendish subgroup  “Grand  Nain”).  In  Panama,  the  sweet  bananas  come  mostly  from  the Cavendish subgroup. The plantain subgroup is also triploid but has the genome group AAB (Simmonds, 1962).

1.1.4 Description of Musa paradisiacae plant

The common  Musa paradisiacae  has broad, irregular  oval leaves,  abruptly contracted at the base into a long broad, channelled footstalk. The fully grown blade is

1.3–2.4  meters  long  and  about  two  third  as broad,  usually  smooth,  with  several

parallel veins. It is wind pollinated and propagates primarily by seeds which are held on the long narrow spikes which rise well above the foliage (Zeller, 2005).

Musa x paradisiaca (M. acuminata x M. balbisiana) is a sterile (without seeds or  viable  pollen)  triploid  (2n=3x=33  chromosomes)  that  is  cultivated  in  warm climates for its tasty yellow-skinned fruit (Nelson et al., 2006). This is a large, fast- growing, suckering, herbaceous perennial that produces huge oblong to paddle-shaped leaves that grow to as much as 8’ long with leaf sheaths overlapping to help form a trunk-like pseudo stem (false stem). The pseudo-stem can reach up to 2-9 m tall and with short underground stem (corm) with buds, from which short rhizomes grow to produce a clump of aerial shoots (suckers)  close to the parent plant. The roots are adventitious, spreading 4-5 m laterally, descending to 75cm long, but mainly in the top of 15cm and form a dense mat. It develops from the underground rhizome (Gibert,

2009).

At maturity, the rhizome gives rise to flower (inflorescence) that is carried up along a smooth elongated unbranched stem piercing through the centre of the pseudo- stem, finally emerging out at the top in between the leaf cluster. Yellow flowers with purple-red bracts appear in summer on mature plants. The flower subsequently develops to plantain bunch consisting of 3 to 20 hands each with at least 5-10 fingers (fruits) (Zeller, 2005). The plant is also monocarpic, which means that a shoot can only flower once and will die after the fruit is produced. The leaf crown will be oriented downward due to gravity.

Raw green fruits are only eaten after cooking. Each fruit measures about 3 to

10 inches or more in length depending on the cultivar type. They tend to have coarse external features with prominent edges and flat surfaces. The flesh inside is starch rich with  tiny  edible  black  seeds  concentrated  at  its  core.  Ripening  process  however enhances flavor and sweetness since the starch converts to  sugar (glucose, fructose and sucrose) (Phebe et al., 2007). The genus honors Antonia Musa, Roman physician of the 1st century B.C.

No serious  insect  or disease  problems.  In some cases,  insects  like aphids,

mealy bugs, moths, scale, thrips, fruit flies and spider mites may attack the  plant. Susceptible to anthracnose, wilt and mosaic virus (Scott et al., 1970).

1.1.5 Distribution of Musa paradisiacae

The plant is widely distributed  throughout  the tropical  regions of Southeast

Asia and western Pacific regions.

It is native to Southeast Asia, India and Burma through the Malay Archipelago to New Guinea,  America,  Australia,  Samona,  and tropical Africa  (Ahmad  et al.,

2006). However,  the cultivation  is limited  to Florida,  the Canary Islands,  Egypt, Southern Japan, and South Brazil. The top leaders exporting countries of plantain are Ecuador, Colombia, Costa Rica, Guatemala and Honduras. Panama occupies the 6th position. The large diversity that occurred  in plantain has  resulted in a variety of cultivars (Scott et al., 1970).

The  number  of Musa  paradisiacae  cultivated  varieties  (cultivars)  has  been reported to vary from one country to another. Swennen (1990) observed that at least

116 plantain cultivars exist in different parts of West and Central Africa. In Nigeria alone, more than 20 cultivars have been reported, although only a few are important commercially  Swennen  (1990).  Musa  paradisiacae   is  a  major   starch  crop  of importance in the human tropical zone of Africa, Asia, Central and South America. It is undoubtedly one of the oldest cultivated  fruits in West  and Central Africa. It is consumed  as an energy yielding food and desert. It  has been estimated  that Musa paradisiacae and other bananas provide nearly 60 million people in Africa with more than 200 calories (food energy) per day.  Fruits such as Musa paradisiacae  are an important contribution  to the diets of  many low and middle class people in many African settings (Stover and Simmonds, 1987). Bananas and plantains constitute the fourth most important global food commodity (after rice, wheat and maize) grown in more than 100 countries over a harvested area of approximately 10 million hectares, with an annual production of 88 million tonnes (Frison and Sharrock, 1999). The all year round fruiting habit of Musa paradisiacae puts the crop in a superior position in bridging the ‘hunger gap’ between crop harvests. It therefore contributes significantly to food and income security of people engaged in its production and trade, particularly in developing countries. Musa paradisiacae is an important staple crop, supplying up to 25% of the carbohydrates for approximately 70 million people in the humid zone of sub-Saharan Africa. (IITA, 1998).

1.1.6 Cultivation and storage of Musa paradisiacae

Musa  paradisiacae  is  grown  in  52  countries  with  world  production  of  33 million metric tonnes (FAO, 2005). It grows more than any other plant in compacted soils,  is  abundant   beside  paths,  roadside   and  other  areas  with   frequent   soil compaction.  It  is also  common  in  grassland  and  as a  weed  among  crops.  Musa paradisiacae originated in the humid tropics and performs best under warm (27-30ºC) and  very wet  (200-220mm  per  month)  conditions.  The  musa  cultivars  can  stand warmer and drier climates (Gibert, 2009). The best soils are deep, friable loam with a good  drainage  and  aeration.  High  soil  fertility  and  organic  matter  content  are desirable. The crop tolerates PH values of 4.5-7.5. It is sensitive to typhoons which cause blow-downs. A major problem of Musa paradisiacae is that the fruits are highly perishable (Scott et al., 1971).  The most important physiological function affecting product quality during storage is respiration and transpiration. To extend storage life, these  functions  should  be  reduced.  This  can  be done by controlling  temperature, humidity,   ventilation,   and  atmospheric   composition   during  storage  (Scott  and Gandanegara, 1974).

1.1.7 Historical uses of Musa paradisiacae

Every part of Musa paradisiacae including root system is used widely in various treatments. The fruit of unripe Musa paradisiacae is traditionally used in the treatment  of diarrhoea,  dysentery,  intestinal  lesions  in ulcerative  colitis,  diabetes (unripe), in sprue, uraemia, nephritis, gout, hypertension, cardiac disease (Mwangi et al., 2007).

Unripe bananas and plantain fruits are astringent, and used to treat diarrhoea.

The leaves are used for cough and bronchitis. The roots can arrest haemoptysis and posses strongly astringent, and antihelmintic properties. Plantain juice is used as an antidote for snakebite. Other uses are asthma, burns, diabetes, dysentery, excessive menstrual   flow,  fever,  gangrene,   gout,  headache,   haemorrhage,   inflammation, insomnia, intestinal parasites, sores, syphilis, tuberculosis, ulcers, and warts (Coe and Anderson, 1999). In Suriname’s traditional medicine, the red protecting leaves of the bud was used  against  heavy menstrual  bleeding  (menorrhagia).  Other  therapeutic uses were against dysentery, migraine, hypertension, asthma and jaundice.

1.1.8 Health benefits of Musa paradisiacae

          Indeed,  they are very reliable sources of starch and energy ensuring  food security for millions of households worldwide (Swennen, 1990).

        It contains dietary fibre. Adequate amount of Dietary-fibre in the food helps normal bowel movements, thereby reducing constipation problems.

        Musa paradisiacae is rich in vitamin C. Consumption of foods rich in vitamin- C helps the body develop resistance against infectious agents  and scavenge harmful oxygen-free radicals.

        Musa  paradisiacae  contains  enough  of vitamin  A.  In addition  to being  a powerful  antioxidant,  vitamin  A  plays  a  vital  role  in  the  visual  cycle, maintaining healthy mucus membranes, and enhancing skin complexion.

     As in bananas, they too are rich sources of B-complex vitamins, particularly

high  in  vitamin-B6  (pyridoxine).  Pyridoxine  is  an  important  B-complex vitamin that has a beneficial role in the treatment of neuritis, anaemia, and to decrease homocystine (one of the causative factors for coronary artery disease (CHD) and stroke episodes) levels in the body. In addition, the fruit contains moderate levels of folates, niacin, riboflavin and thiamine (Ogazi, 1996).

        They also provide adequate levels of minerals such as iron, magnesium, and phosphorous.  Magnesium  is  essential  for  bone  strengthening  and   has  a cardiac-protective role as well.

Musa  paradisiacae   are  also  rich  in  potassium.  Potassium  is  an   important component of cell and body fluids that helps control heart rate and  blood pressure, countering negative effects of sodium (Ogazi, 1996).

1.1.9 Ripening Process and the Chemical Composition of Musa paradisiacae

The chemical composition of Musa paradisiacae varies with variety, maturity, degree of ripeness and where it is grown (soil type). During the  ripening process, Musa paradisiacae  produce  the gas  ethylene,  which acts as a  plant  hormone and indirectly affects the flavour. Among other things, ethylene stimulates the formation of  amylase, an  enzyme that breaks down starch into  sugar, influencing the taste of

bananas (Swennen, 1990).  The greener, less ripe Musa paradisiacae contain higher levels of starch and, consequently,  have a “starchier” taste. On the other  hand, ripe ones taste sweeter due to higher sugar concentrations. Furthermore, ethylene signals the production of  pectinase, an enzyme which breaks down the  pectin between the cells of the banana, causing the banana to soften as it ripens. The water content in the green plant is about 61% and increases on ripening to  about 68%. The increase in water is presumably due to the breakdown of carbohydrate during respiration. Green Musa paradisiacae contains starch which is in the range 21 to 26% (Jaffe et al., 1963; Marriott and Lancaster, 1983). The  starch in the unripe plantain consists of mainly amylose and amylopectin in a ratio of around 1:5. Sugars comprise only about 1.3% of the total dry matter in unripe plantain, but this rises to around 17% in the ripe fruit (Ogazi, 1996). During ripening, the sugars are in the approximate ratio of glucose, 20: fructose, 15: sucrose, 65. Only traces of other sugars are found (Swennen, 1990).  The fat content of plantains is very low, less than 0.5% and so fats do not contribute to the energy content (Jaffe et al., 1963; Marriott and Lancaster, 1983).

The protein content of unripe fruit is between 0.5 and 1.6% and no significant change in the ripening fruit has been detected. The amino acid component includes alanine  amino-butyric  acid,  glutamine,  asparagine,  histidine,  serine,  arginine,  and leucine. The ascorbic acid content is high.  Although the total lipid content remains essentially  unchanged  during  ripening,  the  composition  of  fatty  acids,  especially within the phospholipid  fractions has  been observed  to change, with a decrease in their saturation (Ogazi, 1996).



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ANTIDIARRHOEAL EFFECT OF UNRIPE MUSA PARADISIACAE PULP AND PEEL HOMOGENATES ON CASTOR OIL-INDUCED DIARRHOEA IN WISTAR ALBINO RATS

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