Effects of Carica Papaya Leaf Extract on Blood Hematology, Serum Biochemistry and Immune Response of Broilers

Full Length Research Article

Effects of Carica Papaya Leaf Extract on Blood Hematology, Serum Biochemistry and Immune Response of Broilers

Muhammad Khalid Bashir1, Muhammad Ashraf2, Shahid Ur-Rehman3*, Sadia Razzaq2, Muhammad Qamar Bilal3, Sayeda Mariam4, Madiha Tabbasum5

Adv. life sci., vol. 7, no. 4, pp. 252-256, August 2020
*- Corresponding Author: Shahid Ur-Rehman (Email: shahidurrehman@uaf.edu.pk)
Authors' Affiliations

 1. Directorate of Graduate Studies, University of Agriculture, Faisalabad – Pakistan
2. Department of Poultry Science, University of Agriculture, Faisalabad, Sub Campus Toba Tak Singh –Pakistan
3. Institute of Animal and Dairy Sciences, University of Agriculture, Faisalabad – Pakistan
4. Department of Statistics University of Agriculture, Faisalabad – Pakistan
5. Department of Computer Science University of Agriculture, Faisalabad, Sub Campus Toba Tek Singh – Pakistan
 [Date Received: 04/04/2020; Date Revised: 21/08/2021; Date Published Online: 31/08/2020]

Abstractaa download_button
Introduction
Methods
Results
Discussion
References 

Abstract

Background: As an alternative of antibiotic growth promoters, medicinal plants are the most popular options. Papaya leaves are good source of vitamins, minerals, antioxidants and enzymes which provide aid in digestion of carbohydrates, proteins and fats and overcome the microbial load and used as a growth promoter and antibiotic in broiler performance. This study was carried out in order to evaluate the effect of papaya leaf extract in drinking water as a replacement of commercial antibiotic on blood hematology, serum biochemistry and immune response of broilers for the duration of 35 days.

Methods: In this experiment, 180 one-day old chicks were randomly distributed into 6 treatments. Each treatment was divided into 3 replicates having 10 chicks each. Feed was offered ad-libitum to broilers. Six treatments; G1 {(basal diet + antibiotic growth promoter (enramycin) + coccidiostat (salinomycin)}, G2 (basal diet without antibiotic growth promotor and coccidiostat), G3 (basal diet + 0.5% papaya extract), G4 (basal diet + 1.0% papaya extract), G5 (basal diet + 1.5% papaya extract) and G6 (basal diet + 2.0% papaya extract) were designed. Blood samples were taken for serum biochemistry, hematology and titer against Newcastle disease. Data were analyzed using analysis of variance technique by completely randomized design using SAS and Tukey’s test were used for comparison of means.

Results: Birds received various level of papaya extract had no effect on serum biochemistry and hematology parameters except triglyceride levels which were significantly lower in G2 (15.00±15.00, commercial diet) and G6 (17.43±9.87, commercial diet + 2% papaya extract) at 17th day and 35th day where it failed to show significant difference. However, Newcastle disease and infectious bursal disease titers were not affected by the treatments.

Conclusion: It can be concluded that papaya leaf extract may be used in broiler diet as commercial antibiotics replacement.

Keywords: Papaya leaf extract; Antibiotics; Immune response; Broiler

Introduction6th button-01

Substances that are supplemented in poultry diet to destroy or inhibit growth of microbes especially bacteria are classified under the category of additives [1]. Many of synthetic drugs and growth promoters are used in poultry diets e.g. neomycin, bacitracin manganese, soframycin, tylosin, hygromycin-B, enramycin and erythromycin etc. [2]. However, their excess use develops resistance in bacteria and has adverse effects on human health [3]. Scientific research evidence proposes to reduce use of antibiotic growth promotors because uncontrolled use of these chemicals has increased issues related with antibiotic resistance and antibiotic residues having direct effect on and human and animal wellbeing [4-7]. As an alternative of antibiotic growth promoters, medicinal plants are most popular options. Medicinal plants are cheap and contain pharmacologically active substances which produce some specific chemicals that reduce growth of bacteria [8].

Carica Papaya is a species which belong to Caricaceae family. Its fruit is Papaya which is native fruit of America. It was firstly cultivated in Mexico and then cultivated in other countries [9]. Carica papaya seeds possess moisture 71.89, protein 8.4, carbohydrate 9.44, ash 1.47%, fatty oil-lauric 0.13, myristic 0.16, palmitic 15.13, stearic 3.61, arachidic 0.87, behenic 0.22, oleic 71.60, linoleic 7.68, linolenic 0.6%, Phospholipids- phosphatidylcholine, cardiolipin. Papaya leaves contain high crude protein and low fiber content. It contains about 30% crude protein only 5% crude fiber. It also contains calcium, potassium, magnesium, sodium and iron [10]. Papaya leaves and seeds are good source of vitamins, minerals, antioxidants and enzymes which provide aid in digestion of carbohydrates, proteins and fats and overcome the microbial load and used as a growth promoter and antibiotic in broiler performance [11]. Onyimonyi and Onu found that a 2% inclusion of papaya leaf meal in finishing broilers feed could improve performance, carcass and organoleptic indices [12].

Papaya leaves extract contain phytochemicals which have been reported to improve the platelet and red blood cell [13]. In the treatment of arthritis and intestinal worms, papain (a product which is produced from papaya fruit which mainly contains enzymes) is widely used. Because of the presence of phytochemicals, papaya is widely used to enhance immune system and release natural chemicals which attack on tumor cells [14]. Dried papaya leaves are medically important as a tonic and blood purifier [11].

Papaya leaf extract administrating in drinking water can be replaced with antibiotics without compromising hematology, antibiotic titer and serum biochemistry. Therefore, present study was conducted to study the influence of papaya leaf extract on hematology, serum biochemistry and immune response of broilers.  

Methods6th button-01

Shed preparation

Shed was cleaned, whitewashed and wet cleaned. All holes in shed were closed in order to stop entering of rodents. One-inch layer of saw dust was spread in all pens. Bulbs were checked for proper light intensity in all pens. Brooder was arranged in order to maintain temperature. Shed was sealed properly (not to compromise ventilation) by polythene sheet in order to maintain temperature at brooding. Shed was preheated 24 hours before arrival of chicks. All fans and bulbs were checked before the flock arrival in order to remove any fault if present. Drinkers and feeders were washed and dried in sunshine, then placed in every pen before the arrival of flock. Chicks were received in chick boxes and placed in pens. Birds were weighed and counted. Glucose solution (4%) was supplemented in drinking water to chicks in order to ensure quick supply of energy. Brooding paper was spread over saw dust and feed was spread on it. Temperature and humidity were maintained according to the standard requirements. Feed was offered ad libitum to the birds. Clean and fresh water was provided to chicks.

Preparation of Papaya extract

Fresh papaya leaves were collected, washed and dried under shade and sliced. After grinding 5 g papaya leaf powder was mixed with 1000 ml of water and then boiled for two to three hours at constant 80°Ctemperature.

Bird Husbandry

In this experiment, 180 one-day old chicks were randomly distributed to 6 treatments. Each treatment was divided into 3 replicates having 10 chicks in each. Diets were offered ad-libitum to birds. Six treatments as shown in Table 1 were randomly allotted to the replicates the composition of commercial diet is given in Table 2.

Blood samples

Blood sample with anticoagulant (EDTA; 1 mg/ml) was collected (3-4 ml) from each bird at 17th and 35th day. Serum samples were separated and stored at -20°C. Blood samples were used for different parameters including red blood cell (RBC) count, hemoglobin concentration (Hb%), hematocrit percent (Hc), total leukocyte count (TLC) and differential leukocyte count (DLC) [15]. Serum biochemical parameters were estimated spectrophoto-metrically according to methods described by Ghaffar et al., [16] using commercially available Biomed 122-120, 122-190 and 122-220 kits.

Statistical analysis

Data were analyzed using analysis of variance technique by completely randomized design (CRD) using SAS [17] and Tukey’s [18] test was used to compare the means. 

Results6th button-01

White blood count (109/l) of treatments PC, NC, 0.5, 1.0, 1.5 and 2.0% was 69.7, 96.4, 76.5, 92.6, 96.3 and 80.4, respectively. Lymphocyte (109/l) content of treatments PC, NC, 0.5, 1.0, 1.5 and 2.0% was 61.7, 69.5, 50.5, 69.8, 70.0 and 58.2, respectively. Cells other than Granulocyte or lymphocyte (109/l) of treatments PC, NC, 0.5, 1.0, 1.5 and 2.0% were 15.39, 15.24, 20.56, 16.48, 19.30 and 23.94, respectively. Granulocyte content (109/l) of treatments PC, NC, 0.5, 1.0, 1.5 and 2.0% were 8.56, 14.27, 6.68, 11.59, 9.53 and 9.34, respectively. Lymphocytes percentage of treatments PC, NC, 0.5, 1.0, 1.5 and 2.0% were 61.70, 69.47, 50.47, 69.77, 70.00 and 58.17, respectively. Cells other than Granulocyte or lymphocyte percentage of treatments PC, NC, 0.5, 1.0, 1.5 and 2.0% were 24.30, 15.77, 24.37, 17.70, 20.13 and 30.03, respectively. Granulocyte percentage of treatments PC, NC, 0.5, 1.0, 1.5 and 2.0% were 14.00, 14.80, 7.87, 12.53, 9.87 and 11.83, respectively. RBC content (1012/l) of treatments PC, NC, 0.5, 1.0, 1.5 and 2.0% was 1.9, 2.0, 1.6, 2.2, 2.1 and 2.1, respectively (Table 3).

Hemoglobin content (g/dl) of treatments PC, NC, 0.5, 1.0, 1.5 and 2.0% were 12.13, 13.20, 10.67, 13.87, 13.47 and 13.33, respectively. Hematocrit percentage of treatments PC, NC, 0.5, 1.0, 1.5 and 2.0% were 28.02, 29.91, 23.34, 31.75, 29.91 and 29.95, respectively. Mean Corpuscular Volume (fl) of treatments PC, NC, 0.5, 1.0, 1.5 and 2.0% were, 147.67, 147.33, 147.33, 147.00, 140.33 and 141.67, respectively. Mean Corpuscular Hemoglobin (+pg) of treatments PC, NC, 0.5, 1.0, 1.5 and 2.0% were, 64.00, 65.23, 67.87, 64.37, 63.20 and 63.03, respectively. Mean Corpuscular Hemoglobin Concentration (g/dl) of treatments PC, NC, 0.5, 1.0, 1.5 and 2.0% were, 43.33, 44.17, 46.07, 43.90, 44.97 and 44.47, respectively. Red cell distribution width (+fl) of treatments PC, NC, 0.5, 1.0, 1.5 and 2.0% were, 69.67, 73.57, 67.90, 69.33, 62.17 and 67.17, respectively. Red cell distribution width percentage of treatments PC, NC, 0.5, 1.0, 1.5 and 2.0% were, 10.40, 10.90, 10.20, 9.70, 10.13 and 10.73, respectively (Table 3).

Platelet count (109/l) of treatments PC, NC, 0.5, 1.0, 1.5 and 2.0% were, 11.33, 22.67, 12.00, 20.00, 18.67 and 9.33, respectively. Platelet percentage of treatments PC, NC, 0.5, 1.0, 1.5 and 2.0% were, 0.01, 0.01, 0.01, 0.01, 0.01 and 0.01, respectively. Mean Platelet Volume (-fl) of treatments PC, NC, 0.5, 1.0, 1.5 and 2.0% were, 6.90, 6.73, 6.70, 7.03, 7.10 and 7.00, respectively. Platelet distribution width (-fl) of treatments PC, NC, 0.5, 1.0, 1.5 and 2.0% were, 5.93, 6.40, 5.60, 6.83, 7.03 and 4.70, respectively. Platelet distribution width percentage of treatments PC, NC, 0.5, 1.0, 1.5 and 2.0% were, 29.00, 30.33, 28.57, 30.80, 32.03 and 25.63, respectively. Platelet Large Cell Count (109/l) of treatments PC, NC, 0.5, 1.0, 1.5 and 2.0% were, 1.00, 2.33, 1.33, 3.00, 2.00 and 1.00, respectively. Platelet Large Cell Ratio percentage of treatments PC, NC, 0.5, 1.0, 1.5 and 2.0% were, 9.42, 11.25, 9.83, 9.92, 9.83 and 10.27, respectively. Lyse content (ml) of treatments PC, NC, 0.5, 1.0, 1.5 and 2.0% were, 0.90, 0.90, 0.90, 0.90, 0.90 and 0.90, respectively (Table 3).

At 17th day, highest triglycerides concentration (105.67±24.07) was recorded in the blood of group G3 while lowest concentration (15.00±15.00) was noted in blood of group G2 birds. Cholesterol level was highest (161.00±19.65) in group G4 and lowest (83.87±42.93) in group G3. Group G3 also showed highest high density lipoprotein (HDL) level (39.72±4.42) and group G5 recorded lowest (34.45±1.83) HDL. Highest Low density lipoprotein (LDL) concentration (109.92±17.85) was recorded in group G4 and lowest LDL concentration (23.01±44.07) was seen in group G3. Statically analysis of data revealed that blood chemistry parameters, (triglycerides, cholesterol, HDL and LDL) were non-significantly (P>0.05) different (Table 4).

At 35th day, group G5 revealed highest triglycerides concentration (80.53±20.55) while group G6 showed lowest concentration (17.43±9.87). Cholesterol level was also highest (173.33±10.41) in group G5 and lowest (101.67±13.17) in group G3. Group G6 had highest HDL level (30.34±0.87) and group G2 had lowest (24.90±1.28) HDL. Group G5 revealed highest LDL concentration (129.03±7.84) in blood while group G3 had lowest LDL concentration (60.66±17.29). Statically analysis of data revealed that blood chemistry parameters, (triglycerides, cholesterol, HDL and LDL) were non-significantly (P>0.05) different (Table 5).

Antibodies titer against Newcastle disease were also assessed to study the effect of papaya leaves’ extract on immune system of the broilers of treatments positive control (PC), negative control (NC), 0.5, 1.0, 1.5 and 2.0% was 2.33, 1.67, 3.33, 3.33, 3.00 and 3.33 at 17th day and 5.67, 5.67, 5.33, 6.00, 5.33 and 6.00 at 35th day, respectively. Antibodies titers against Newcastle disease were non-significantly different (P>0.05) in birds given various levels of papaya leaf extract in drinking water (Table 6).

Figures & Tables


 

 

 

 

 

 

 

 

 

 

Discussion6th button-01

Blood hematology parameters were non-significantly (P>0.05) different in broiler birds received papaya extract in drinking water. Oloruntola in 42-day feeding trial reported similar results of feeding the effects of a diet supplemented with pawpaw leaf and seed meal composite mix (PCM) on hematological indices [19]. White blood cell counts of birds fed PCM supplemented diets were similar/comparable to that of the birds fed control diet. Hemoglobin concentration and packed cell volume of birds fed PCM supplemented diets were similar to those fed control diet, except for those fed diet 1.0% PCM had significantly (P<0.05) lower Hbc and PCV. Similar results were noted by Mahejabin et al., who observed that papaya extract had no effect on blood parameters [20]. Results are not in line with the previous study who found that papaya leaf had higher total erythrocyte count and erythrocyte sedimentation rate. Similarly, Ogbuokiri et al., showed that higher PVC, lower RBC, higher Hb and lower Neutrophil were recorded in birds received 2.5% Papaya leaf [22]. Ameen et al., also found that birds received Carica papaya extract had higher packed cell volume, haemoglobin concentration, lymphocyte counts, red blood cells and lower eosinophil counts [23]. Contrary results were reported by Sorwar et al., who found that papaya leaf had higher total erythrocyte count (2.57±0.23 million/mm3) and erythrocyte sedimentation rate (6.13±0.64 mm in first hour) [21].

Papaya extract had no effect on blood chemistry parameters (triglycerides, cholesterol, HDL and LDL) and ND titer. Oloruntola et al., reported the effect of pawpaw leaf meal inclusion and enzyme supplementation in the diet of broiler chickens. Enzyme reduced (p < 0.05) low-density lipoproteins (LDL), whereas PLM reduced (p < 0.05) cholesterol and LDL [24]. Similar results were noted by Hussain et al., who observed that papaya extract had no effect on blood parameters line total erythrocyte count, hemoglobin, pack cell volume and erythrocyte sedimentation rate [15].

Present study concluded that papaya extract can be used in drinking water of poultry without any adverse effect on hematology, antibiotic tire and serum biochemistry.

Authors' Contribution

Muhammad. Khalid Bashir, Muhammad Ashraf, Sadia Razzaq, Shahid Ur Rehman and Muhammad Qamar Bilal planned, conducted and took data for the trial. Muhammad. Khalid Bashir, Muhammad Ashraf, Sadia Razzaq, Shahid Ur Rehman and Muhammad Qamar Bilal written and refined the manuscript. Sadia Razzaq, Sayeda Mariam and Madiha Tabbasum analyzed the data statistically.

Conflict of interest

The authors declare that there is no conflict of interest regarding the publication of this paper.

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