Indian Journal of Animal Research

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Research Article

Indian Journal of Animal Research, volume 54 issue 7 (july 2020) : 874-878

Comparative effects of different concentrations of garlic (Allium sativum) and ginger (Zingiber Officinale) on growth performance, goblet cell histochemistry and gut microbiota of broilers

Muhammad Usman Saleem1,*, Muhammad Arshad Javid1, Saleem Akthar2, Faisal Ayub Kiani2, Omer Naseer3, Muhammad Yasir Waqas4
1Department of Bio-Sciences, Faculty of Veterinary Sciences, Bahauddin Zakariya University, Multan, Pakistan.
2Department of Clinical Sciences, Faculty of Veterinary Sciences, Bahauddin Zakariya University, Multan, Pakistan.
3Department of Clinical Medicine, Cholistan University of Veterinary and Animal Sciences, Cholistan, Pakistan.
4Department of Physiology and Biochemistry, Cholistan University of Veterinary and Animal Sciences, Cholistan, Pakistan.
Cite article:- Saleem Usman Muhammad, Javid Arshad Muhammad, Akthar Saleem, Kiani Ayub Faisal, Naseer Omer, Waqas Yasir Muhammad (2019). Comparative effects of different concentrations of garlic (Allium sativum) and ginger (Zingiber Officinale) on growth performance, goblet cell histochemistry and gut microbiota of broilers . Indian Journal of Animal Research. 54(7): 874-878. doi: 10.18805/ijar.B-1105.

ABSTRACT

The present study investigates comparative effects of different concentrations of garlic (Allium sativum) and ginger (Zingiber Officinale) on growth performance and selected histo-microbial parameters of broilers.  A total of 80 one day- old chicks were divided into five different groups with each group having 4 replicates in a trial of 42 days. The first group was kept as control whereas, the second, third, fourth and fifth groups were given garlic 0.25%, garlic 0.5%, ginger 0.25% and ginger 0.5% respectively. At the end of trial all birds were slaughtered for analysis. Results revealed that feed conversion ratio and live body weight were significantly (P < 0.05) improved by garlic 0.5% supplementation compared to other groups. Histo-microbiology revealed that lactic acid bacteria, yeast, acidic goblet cells, mixed goblet cells and total goblet cells increased significantly (P < 0.05) by the dietary supplementations under study. In was concluded that 0.5% garlic supplementation was a better alternate to antibiotic in broilers.

INTRODUCTION

Antibiotic growth promoters (AGPs) were used extensively in the last few decades to increase the production performance and profitability of broilers (Abudabos et al., 2018). Prolonged misuse and overuse of AGPs caused harmfulness to human health (Min et al., 2016) as a result of which the European Union in 2006 has banned the use of AGPs (Tehseen et al., 2016). This ban has negatively affected profitability and performance of broilers (Manafi et al., 2015) which has triggered scientists to find alternative to AGPs for improving growth performance of broilers by optimizing their gut health (Junaid et al., 2018). Poultry diet contains a variety of additives and supplements (Hashemi et al., 2014) among which phytobiotics are being investigated by many scientists for being “Natural Safe Additives” (Boka et al., 2014).
        
Garlic (Allium sativum) belonging to family Amaryllidacea has been reported to alter the microarchitecture of intestine resulting in better growth performance and feed conversion ratio (FCR) (Oladele et al., 2012). It is effective in reducing the number of pathogenic bacteria residing in the intestine of birds that are responsible for causing many pathological conditions (Peinado et al., 2013). It is added in feed due to exhibiting antifungal, antibacterial, antioxidative, (Saeid et al., 2013) immunomodulatory and antiparasitic properties. Garlic contains allin, diallylcysteine, allicin, dithiin, ajoene and S-allylcysteine (Rehman and Munir, 2015).
        
Ginger (Zingiber Officinale) belongs to family Zingiberaceae (Khonyoung et al., 2017) and is rich in trace minerals and essential oils (Ogbuewu et al., 2017). Gingerdiol, zingeronen zingibrene, shogaols and gingeroiols are present in ginger giving it anti-inflammatory, hepatoprotective, analgesic, antioxidant, cardioprotective, immunomodulatory, neuroprotective, antioxidant and antimicrobial properties (Herve et al., 2018). The proximate composition of fresh ginger is 2.3% protein, 12.3% carbohydrates, 1% fat, 2.4% fiber, 80.8% water and 1.2% ash whereas, dried ginger has 10% moisture (Muhammad et al., 2017). To the best of our knowledge no data is present regarding comparative effects of different concentrations of garlic and ginger on growth performance, goblet cell (GC) histochemistry and gut biome of broilers. This study therefore aims at investigating the comparative effects of garlic and ginger, in different concentrations on growth performance and selected histo-microbial parameters of broilers.

MATERIALS AND METHODS

This study was carried out in environmentally-controlled broiler shed. The experiment lasted for 42 days and was conducted on 80 day-old Hubbard broiler chicks obtained from a commercial hatchery. Upon arrival birds were weighed and randomly assigned to five groups with each group having four replicates. The number of birds in each replicate were four (n=4). The first group (CONT) was kept as control and was fed basal diet (Table 1). The second group (GAS 0.25%) and third group (GAS 0.5%) were supplemented with garlic (Garlic - Fooding group Ltd, Shanghai, China, 0.25gm/kg and 0.5gm/kg of basal diet) respectively whereas, the fourth (GZO 0.25%) and fifth (GZO 0.5%) group was given ginger (Garlic - Fooding group Ltd, Shanghai, China, 0.25gm/kg and 0.5gm/kg of basal diet) respectively. Birds were vaccinated intraocularly by live attenuated Newcastle disease virus (Ceva-Phylaxia, Budapest, Hungary) on day 1, with a booster on day 21 in drinking water. Similarly vaccination against infectious bursal disease (Lohman Animal Health GmbH, Cuxhaven, Germany) was done by intraocular route on day 8 and repeated on day 20 in drinking water.
 

Table 1: Composition of experimental diet given to broilers (Saleem et al., 2019).


 
Growth performance and relative organ weight
 
During the whole experiment feed and water to birds of each replicate were provided ad-libitum and their refusals were weighed daily. Feed intake was recorded weekly. At the end of the experiment all the birds were slaughtered. Feed conversion ratio was estimated as feed consumed by each bird in grams divided by weight gain per bird.
 
Histochemistry of goblet cells and intraepithelial lymphocyte count
 
About 3 cm long small intestinal segments from midpoints of duodenum (segment encompassing the duodenal loop), jejunum (segment between duodenum and ileum) and ileum (distal segment before the ileo-cecal junction equaling the length of caecum) were taken and fixed in 10% neutral buffered formalin. Segments were then embedded in paraffin. Slides prepared by paraffin embedding technique were stained by combined alcian blue - periodic acid Schiff (AB - PAS) and observed at 10× for counting goblet cells. Five villi per bird were studied for counting the goblet cells. Goblet cells (GC) were differentiated as acidic and mixed on the basis of staining ability of mucin contents. Acidic goblet cells (AGC) having acidic mucin stained blue whereas, mixed goblet cells (MGC) having both acidic and mixed mucin were stained purple (Saleem et al., 2018).
 
Microbiology of small intestine
 
Intestinal contents from duodenum, jejunum and ileum were immediately collected and kept at 4°C in sterile bags until being plated on agar. Samples were prepared by making dilutions and from the diluted extracts (10-1 to 10-9) plates were prepared for specific organisms under study. For lactic acid bacteria (LAB) MRS agar (Himedia laboratories Pvt. Ltd., Mumbai, India) whereas, malt extract glucose peptone agar (Himedia laboratories Pvt. Ltd., Mumbai, India) for yeast were used. Counts of LAB and yeast after four days of incubation at 30°C were recorded and converted to logarithmic colony forming units (cfu/g) (Saleem et al., 2018).
 
Statistical analysis
 
Analysis of data was conducted with Statistical Package for Social Science (SPSS for windows version 20.0 SPSS, Chicago, USA). Data was found to be normally distributed after checking with Kolmogrov Simirnov test. Data for groups was analyzed with one way analysis of variance (ANOVA). Differences were considered significant at P < 0.05 and were calculated by applying Duncan’s multiple-range test.

RESULTS AND DISCUSSION

Response of broilers to different concentrations of garlic and ginger on growth performance is presented in Table 2. Live body weight and FCR of the birds increased significantly (P < 0.05) by all the dietary treatments under study compared to CONT. Birds of GAS 0.5% had significantly (P < 0.05) high live body weight and FCR compared to the other groups. No significance (P > 0.05) was observed for feed intake of broilers by supplementation of garlic and ginger.
 

Table 2: Effect of garlic (Allium sativum) and ginger (Zingiber officinale) on growth performance of broilers.


        
Addition of phytogenic substances in broiler feed leads to improved weight gain because of the presence of certain compounds and bioactive components that enhance digestibility and absorption of feed (Alagawany et al., 2016). Allicin active ingredients present in garlic improve digestion leading to better weight gain (Eltazi, 2014). Our results of improvement in live weight of broilers after supplementation of garlic are in collaboration with the results of Lewis et al., (2003). Results similar to our findings were reported by Abdullah et al., (2010) that 0.5% garlic supplementation of broilers leads to better weight gain. Supplementation of ginger increases body weight due to the presence of active ingredients that stimulate digestive enzymes that lead to better absorption and digestion of feed (Karangiya et al., 2016). Ginger enhances pancreatic lipase activity, intestinal lipase, disaccharidase, sucrose and maltase activities (Platel and Srinivasan, 2000) all of which have a positive effect on gut function leading to better weight gain of birds. Similar to our results Mohamed et al., (2012) mentioned that adding ginger to broiler diet had a positive significant effect on live body weight. Analogous to our results Onu, (2010) reported that feed intake of birds is not affected by supplementation of garlic and ginger.
        
Birds of all the treatment groups exhibited significant (P < 0.05) increase in AGC, MGC and TGC count compared to CONT in all the segments of small intestine. However no significance was observed in AGC, MGC and TGC by the difference of dietary treatments as shown in Table 3.
 

Table 3: Effect of dietary supplementations on small intestine microarchitecture in broilers.


        
Intestinal epithelium is covered by mucus which is secreted by goblet cells (Rahmatnejad and Saki, 2016). Mucin present in the mucus provides potential binding sites to beneficial bacteria thus decreasing chances of colonization of pathogenic bacteria (Hutsko et al., 2016). Regulation of mucin gene upholds the activities of goblet cells and contributes towards the protective mechanisms of gut (Sikander et al., 2017). Stem cells proliferate in the crypts and move up the villus where they are differentiated as enterocytes and goblet cells. Goblet cells increase by augmented proliferation rate of stem cells in gut (Hutsko et al., 2016). Addition of phytobiotics like garlic and ginger improves intestinal microarchitecture by enhancing intestinal mucosal proliferation activity (Karangiya et al., 2016) which explains our results for increase of AGC, MGC and TGC in all segments of small intestine.
        
Small intestinal microbiology revealed that in all the treatment groups the number of LAB and yeast increased significantly (P < 0.05) in all the segments of small intestine compared to CONT. The number of LAB was found to be significantly (P < 0.05) higher in GAS 0.5% compared to the birds of all the other treatment groups. However yeast was not affected by the difference of dietary supplementations under study as shown in Table 4.
 

Table 4: Effect of garlic (Allium sativum) and ginger (Zingiber officinale) on microbiota of small intestine.


        
Intestinal microbiota is the largest bacterial reservoir in birds having both beneficial and non-beneficial bacteria. When the balance between these two bacterial populations is maintained the birds perform to their maximum, however, any disturbance in this balance affects growth performance of bird negatively (Blajman et al., 2015). The LAB are gram positive, catalase negative, non-sporulating, rod shaped fermentative organisms with lactic acid as major end product of carbohydrate fermentation. They are responsible for protection and stability of gut ecosystem (Hernandez et al., 2016). Yeast has been reported to have antibacterial properties (Hatoum et al., 2012) and provides protection against pathogenic bacteria by (i) acting as a competitive exclusion agent for attachment with enterocytes (ii) secreting enzymes that degrade bacterial toxins (iii) producing mycocins (Caly et al., 2015). Our results are supported by the report of Karangiya et al., (2016) that garlic and ginger supplementation increase the number of LAB in gut of broilers.

CONCLUSION

In light of the above said results it is concluded that addition of 0.25%, 0.5% of garlic and ginger improved growth performance and histo-microbial parameters of gut in broilers. Moreover addition of 0.5% garlic to broilers diet may be a better alternative to antibiotic growth promoters.

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