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Effects of Powdered Balsam Apple (Momordica Balsamina) Leaf Supplement on Blood Glucose Level and the Haematological Parameters of Japaneese Quails

A. H. Jibril U. Musa B. Saidu A. B. Ajape I. H. Maina A. A. Jimoh A. Sani Y.A. Yabo A. I. Jaafar
Vol 8(2), 99-105
DOI- http://dx.doi.org/10.5455/ijlr.20170817041629

This experiment was conducted to assess the effect of Balsam leaf crude extract on blood glucose level and the haematological parameters of Japanese quails. Fourty eight specific pathogen free quails were divided into four (4) categories which include control, groups A, B and C. Control group received no treatment of the crude extract; the groups received 5g, 10g and 15g respectively. Data were subjected to ANOVA to compare means between groups. After the administration of the extract, there was statistically significant (p< 0.05) marked decrease in the blood glucose level, coupled with increase in the leucocytes and erythrocytes counts in a dose and time dependent manner. This indicated that, the plant possess hypoglycemic and haematinic effect which makes it a possible anti-diabetic and haematinic agent of choice. This plant can be used as a supplement/adjunct in the management of type 2 diabetes mellitus.


Keywords : Balsam Apple Erythrocyte Hypoglycemia Haematinic Quails Leucocytes

Introduction

Momordica balsamina leave commonly known as African pumpkin (or African cucumber), Balsam apple (or Balsam pear) and locally called “Garahuni” (Hausa Language), belongs to the family Cucurbitaceae. The plant is perennial herb with soft stems and tendrils that climb up shrubs, boundary fields and fences (Welman, 2004). It is a wonder plant for neutraceutical sciences. The leaves are important source of nutrients having 17 amino acids (Hassan and Umar, 2006), adequate mineral composition like potassium, magnesium, phosphorus, calcium, sodium, zinc, manganese and iron contributes towards combating the problem of micronutrient deficiencies in soil and high value of protein and fat with low fibre content (Hassan and Umar, 2006; Flyman and Anthony, 2007). The wild vegetable could be promoted as a protein supplement for cereal-based diets in poor rural communities and its high potassium content is a good source for the management of hypertension and other cardiovascular conditions. The leaves, fruits, seeds, and bark of these plants are reported to have various medicinal and nutritional properties (Hassan and Umar, 2006; Benoit-Vcal et al., 2006; Bot et al., 2007). The fruit pulp extract of Momordica balsamina shows anti-HIV property (Bot et al., 2007). ‘Momordins’ present in the plant is capable of inhibiting the growth of HIV and other viruses (Ortiago and Better, 1992). Recent studies conducted by Jibril et al. (2015) had showed the efficacy of Balsam apple leaf in the management of Newcastle in Zamfara State, Nigeria. This study aimed at determining the effect of the plant extract on the hematological parameters and blood glucose level in Japanese quails.

Materials and Methods

Preparation of Crude Plant Extract

Balsam apple leaf used for this study was collected at Nasarawa village of Jega Local Government area Kebbi State, Nigeria and transported to the Botany Unit of the Department of Biological Sciences, Usmanu Danfodiyo University, Sokoto, for botanical identification.  Methods described by Abuye et al., 2003 was adopted to manually washed with distilled water and residual moisture was evaporated at room temperature and dried in oven in a paper envelope at 550C for 24 hours. The dried crude extract was grounded into fine powder with the aid of pestle and mortar, and sieved through mesh sieve. Proximate analysis was conducted on the crude extract at the research laboratory of Faculty of Agriculture, Usmanu Danfodiyo University, Sokoto to evaluate the moisture, ash, lipid, crude fibre, nitrogen, crude protein and mineral contents of the plant using the official methods of AOAC, 1990.

Feeding Regimen

All the groups had access to drinking water and feed ad libitum. They were fed with commercial poultry feed (Vital Feed® Nigeria) under suitable environmental condition.  Feed consumption of each quail per day was estimated using the following formula-

 

 

Experiment Protocol

Forty eight week old apparently healthy Japanese quails, weighing between 100 – 120g were obtained from a breeder in Sokoto State metropolis. They were conditioned for one week before the commencement of the study. The quails were divided into four groups of four birds each in a cage made up of zinc and wire mesh. The selected doses was based on the study of Adewale et al., 2014 that administered 12.5g, 25g and 50g as a low, moderate and high dose of aqueous leaf extract of Momordica charantia in wistar rats. The quails were divided according to the schedule below-

Control: Quails in this group were the control and were fed on normal           poultry diet throughout the experiment period.

Group A: 5g of the crude plant extract was incorporated into the total quantity of feed consumed per quail in the group.

Group B: 10g of the crude plant extract was incorporated into the total quantity of feed consumed per quail in the group.

Group C: 15g of the crude plant extract was incorporated into the total quantity of feed consumed per quail in the group.

Meteorological Parameters

The daily values for ambient temperature (AT) and relative humidity (RH) were collected using Digital temperature hygrometer, BIOBASE ® China. These parameters were recorded daily during the morning (08:00-10:00 h) and afternoon (13:00-16:00 h) throughout the study period (early May-ending June). The temperature humidity index (THI) was calculated using the following equation (Ravagnolo et al., 2000).

THI = (1.8 × T + 32) – [(0.55 – 0.0055 × RH) × (1.8 × T -26)]

where T = ambient temperature (° C) and RH = relative humidity (%)

Blood Sample Collection

Blood samples were collected via jugular vein puncture from four birds, in each groups and transferred into an EDTA bottles. The blood samples were transported on ice pack to Haematology Laboratory of the Department of Veterinary Physiology and Biochemistry, Usmanu Danfodiyo University for hematological analysis. This was repeated every week for a period of three weeks.

Haematology and Blood Glucose Determination

Haematological parameters including WBC (using heamocytometer), RBC counts, differential leucocytes counts and Wintrobe indices were determined using methods described by Schalm et al., 1975 and Coles 1974. Haemoglobin was determined using digital colorimeter. Blood glucose level was measured before placing the blood samples into the EDTA bottles using Accu-Chek® Strip and Meter (Roche Diagnostics, Germany).

Data Management and Analysis

Data were expressed as means ± standard deviation. Data were exported and analysed using Microsoft Office Excel ®2007. Variables were analysed using ANOVA assuming unequal variance. Values of p less than 0.05 were considered significant.

Result

The average temperature and relative humidity recorded were 39.5 °C and 38 % respectively during this study. The temperature humidity index was estimated to be 87.7. The mean and standard deviation of the heamatological parameters of quails were recorded for three weeks following administration of different doses of balsam apple leaf. From this experiment, after one week of oral administration of the leave extract, there was a decrease in blood glucose level in a dose dependent manner as compared with the control, groups B and C with higher dose of the extract (10g and 15g) having lower blood glucose level. However, this difference was not statistically significant (p > 0.05).  There was a statistically significant (p < 0.05)  increase in total white blood cells counts in a dose dependent manner, with group C taking highest dose (15g) showing highest total while blood cells count (29.95 x 103/ml) as indicated in Table 1.  The red blood cell counts shows level of statistical insignificance (p > 0.05).

Table 1: Effect of one week administration Balsam apple leaf extract on haematological parameters of Japanese quails

Parameters Control Group A Group B Group C p – values
Mean + SD Mean + SD Mean + SD Mean + SD
n = 4 n = 4 n = 4 n = 4
Glucose (mg/dL) 266.75 + 18.03 267.00 + 9.90 247.50 + 17.68 239.00 + 31.11 0.1
Haemoglobin (mg/dL) 16.80 + 0.91 15.00 + 0.57 18.35 + 0.64 16.25 + 0.35 3.59 x 10-5
PCV (%) 51.25 + 2.50 46.00 + 1.41 56.50 + 2.12 50.50 + 0.71 1.24 x 10-5
WBC Count (x 103/ml) 22.00 + 0.42 24.30 + 0.21 27.08 + 1.03 29.95 + 0.28 1.83 x 10-10
RBC Count (x 106/ml) 2.71 + 0.73 2.27 + 0.11 3.13 + 0.04 2.72 + 0.03 0.27
MCV (g/dL) 199.45 + 52.11 202.74 + 3.87 180.48 + 4.33 185.69 + 4.53 0.59
MCHC (g/dL) 32.78 + 0.58 32.61 + 0.23 64.96 + 0.10 32.18 + 0.25 0.11
MCH (g/dL) 65.35 + 17.11 66.10 + 0.81 58.62 + 1.23 59.75 + 1.92 0.52
Differential Cell Counts
Lymphocytes (%) 80.75 + 2.50 67.50 + 0.71 79.50 + 3.54 69.00 + 9.90 0
Heterophils (%) 17.50 + 3.11 29.00 + 0.00 18.00 + 2.83 27.00 + 7.07 0
Monocytes (%) 2.00 + 0.82 3.50 + 0.71 2.50 + 0.71 4.00 + 2.83 0.17
Eosinophils (%) 0.00 + 0.00 0.00 + 0.00 0.00 + 0.00 0.00 + 0.00 0
Basophils (%) 0.00 + 0.00 0.00 + 0.00 0.00 + 0.00 0.00 + 0.00 0

PCV: Packed Cell volume, MCV = Mean Corpuscular Volume; MCHC = Mean Corpuscular Haemoglobin Concentration; MCH = Mean Corpuscular haemoglobin, n = number of birds; WBC = White Blood Cell; RBC = Red Blood Cell; Control = Receive no extract; Group A = Administered with 5g of extract; Group B = Administered with 10g of extract ; Group C = Administered with 15g of extract

However, after the second and third weeks of oral administration of the extract, there was a marked statistical significant (p < 0.05) decrease in blood glucose level in a dose dependent manner as compared with other groups. There was also a statistical significant increase in total white blood cells and red blood cells counts (p < 0.05) two and three weeks following oral administration of the extract of balsam apple leaf as indicated in Tables 2 and 3.

Table 2: Effect of two weeks administration Balsam apple leaf extract on haematological parameters of Japanese quails

Parameters Control Group A Group B Group C P ­– values
Mean + SD Mean + SD Mean + SD Mean + SD
n = 4 n = 4 n = 4 n = 4
Glucose (mg/dL) 258.50 + 9.54 170.00 + 2.83 169.00 + 7.07 162.50 + 2.12 4.16 x 10-11
Haemoglobin (mg/dL) 17.08 + 0.86 17.70 + 2.55 19.60 + 0.85 19.80 + 2.40 0.06
PCV (%) 51.25 + 2.22 55.00 + 7.07 60.00 + 2.83 61.00 + 7.07 0.03
WBC Count (x 103/ml) 22.18 + 1.06 23.95 + 1.20 27.15 + 1.48 30.85 + 1.34 5.23 x 10-7
RBC Count (x 106/ml) 2.90 + 0.13 3.09 + 0.98 3.56 + 0.02 3.74 + 0.12 0.04
MCV (g/dL) 177.05 + 3.54 183.97+ 35.69 168.76 + 6.95 163.10+ 13.68 0.31
MCHC (g/dL) 33.32 + 0.83 32.15 + 0.49 32.67 + 0.13 32.45 + 0.18 0.03
MCH (g/dL) 58.98 + 2.02 59.06 + 10.56 55.13 + 2.06 52.94 + 4.73 0.26
Differential Cell Counts
Lymphocytes (%) 80.25 + 2.50 77.50 + 3.54 79.00 + 1.41 72.00 + 0.00 0
Heterophils (%) 18.50 + 1.29 21.50 + 3.54 19.00 + 1.41 26.00 + 1.41 0
Monocytes (%) 1.25 + 1.50 1.00 + 0.00 2.00 + 0.00 2.00 + 1.41 0.36
Eosinophils (%) 0.00 + 0.00 0.00 + 0.00 0.00 + 0.00 0.00 + 0.00 0
Basophils (%) 0.00 + 0.00 0.00 + 0.00 0.00 + 0.00 0.00 + 0.00 0

PCV: Packed Cell volume, MCV = Mean Corpuscular Volume; MCHC = Mean Corpuscular Haemoglobin Concentration; MCH = Mean Corpuscular haemoglobin, n = number of birds; WBC = White Blood Cell; RBC = Red Blood Cell; Control = Receive no extract; Group A = Administered with 5g of extract; Group B = Administered with 10g of extract ; Group C = Administered with 15g of extract

Table 3: Effect of three weeks administration Balsam apple leaf extract on haematological parameters of Japanese quails

Parameters Control Group A Group B Group C P – values
Mean + SD Mean + SD Mean + SD Mean + SD
n = 4 n = 4 n = 4 n = 4
Glucose (mg/dL) 236.25 + 14.73 166.00+ 26.87 157.00+ 22.63 140.50+ 36.06 0
Haemoglobin (mg/dL) 17.43 + 0.54 17.50 + 0.57 16.65 + 0.78 18.65 + 1.63 0.03
PCV (%) 52.50 + 1.29 53.00 + 1.41 52.50 + 2.12 56.00 + 5.66 0.14
WBC Count (x 103/ml) 23.25 + 0.68 24.18 + 0.67 27.25 + 1.20 30.90 + 1.70 3.6 x 10-7
RBC Count (x 106/ml) 3.46 + 0.28 2.59 + 0.52 2.74 + 0.45 3.73 + 0.88 0.01
MCV (g/dL) 152.48 + 12.10 209.46 + 47.77 190.2 + 23.22 152.57+ 20.70 0.01
MCHC (g/dL) 33.19 + 0.41 33.02 + 0.19 32.33 + 0.18 33.33 + 0.46 0
MCH (g/dL) 50.56 + 4.06 69.20 + 16.12 61.45 + 7.14 50.90 + 7.64 0.01
Differential Cell Counts
Lymphocytes (%) 82.25 + 3.40 80.00 + 5.66 80.00 + 4.24 80.50 + 2.12 0.77
Heterophils (%) 16.00 + 4.08 18.50 + 4.95 18.50 + 3.54 18.00 + 1.41 0.67
Monocytes (%) 1.75 + 0.96 1.50 + 0.71 1.50 + 0.71 1.50 + 0.71 0.94
Eosinophils 0.00 + 0.00 0.00 + 0.00 0.00 + 0.00 0.00 + 0.00 0
Basophils (%) 0.00 + 0.00 0.00 + 0.00 0.00 + 0.00 0.00 + 0.00 0

PCV: Packed Cell volume, MCV = Mean Corpuscular Volume; MCHC = Mean Corpuscular Haemoglobin Concentration; MCH = Mean Corpuscular haemoglobin, n = number of birds; WBC = White Blood Cell; RBC = Red Blood Cell; Control = Receive no extract; Group A = Administered with 5g of extract; Group B = Administered with 10g of extract; Group C = Administered with 15g of extract

Discussion

This research was conducted in an attempt to assess possible effect of the balsam apple leaf extract on the haematological parameters and blood glucose level of Japanese Quails. A marked decrease was observed in the blood glucose levels. This decrease in a dose and time manner may be due to the alpha glucosidase inhibitory activity of balsam apple leaf extract. Alpha glucosidase inhibitors are those agents that reduce the impact of carbohydrates on blood sugar making it an agent of choice for management of diabetes mellitus type 2 which work by inhibiting carbohydrates metabolism. This is consistent with the finding of Karumi et al. (1999) that stated that the hypoglycemic property of balsam apple leaf extract could be due to inhibition of glucose uptake in the gastro-intestinal tract. There was also an increase in total white blood cell count which might be due to the immune response following the administration of the extract. The immunogenic activity observed may augment the anti-inflammatory properties (Otimenyin and Uguru, 2005) of the herb by reducing or eliminating the causative agents of inflammation and thus decreasing the nociceptive stimulants released from inflamed cell. This implies that the M. balsamina alone will definitely combat the infection and pain associated with infectious and diabetic diseases thus reducing the number of drugs used in pain, infection and diabetes management and preventing the complications of poly pharmacy. There was also increase in red blood cell count which might be due to presence of iron and zinc in the extracts which aided the process of hemopoiesis and enhance maturation of immature erythrocytes.

Conclusion

This experiment, showed the hypoglycemic, immunogenic and haematopoietic efficacies of balsam apple leaf extract in Japanese quail. This may be a potential neutraceutical in the management of diabetes in humans and animals (dog).

Competing Interests

All authors declared that, here is no any competing interest that could influence this manuscript.

Acknowledgement

The authors would like to appreciate the technical support of laboratory technicians in the Department of Veterinary Physiology and Biochemistry of Usmanu Danfodiyo University, Sokoto .

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