Afzan Mat Yus of，Muhammad Lokman Md Isa

1Department of Biomedical Science，Kulliyyah of Allied Health Sciences，International Islamic University Malaysia，Jalan Sultan Ahmad Shah，Bandar Indera Mahkota，Kuantan 25200，Pahang，Malaysia

2Integrated Cellular and Molecular Biology Cluster（iMolec），Integrated Centre for Animal Care and Use，International Islamic University Malaysia，Jalan Sultan Ahmad Shah，Bandar Indera Mahkota，Kuantan 25200，Pahang，Malaysia

Prevalence of gastrointestinal nematodiasis and coccidiosis in goats from three selected farms in Terengganu，Malaysia

Afzan Mat Yus of1，2*，Muhammad Lokman Md Isa2

1Department of Biomedical Science，Kulliyyah of Allied Health Sciences，International Islamic University Malaysia，Jalan Sultan Ahmad Shah，Bandar Indera Mahkota，Kuantan 25200，Pahang，Malaysia

2Integrated Cellular and Molecular Biology Cluster（iMolec），Integrated Centre for Animal Care and Use，International Islamic University Malaysia，Jalan Sultan Ahmad Shah，Bandar Indera Mahkota，Kuantan 25200，Pahang，Malaysia

ARTICLE INFO

Article history:

Accepted 22 May 2016

Available online 26 Jul 2016

Age

Coccidia

Goat

Nematodes

Terengganu

Objective:To reveal the prevalence of gastrointestinal nematodiasis，Trichostrogylus，Haemonchus，Strongyloides and Trichuris and coccidiosis，mainly Eimeria in kids，yearlings and adult goats.

Methods:A total of 287 goat faecal specimens were randomly collected from three farms in Kuala Terengganu，Setiu and Besut.

Results:Prevalence of coccidiosis（89.2%）was significantly higher（P＜0.05）than gastrointestinal nematodiasis（52.3%）.Gastrointestinal nematodes（GINs）were found in 37.0%kids，63.5% year lings and 53.3%adults.Identification of GINs revealed the predominance of Strongyloides（45.6%）followed by Trichostrogylus（12.9%），Haemonchus（11.8%）and Trichuris（8.7%）in all groups whereby the adult goats recorded the highest infections of Trichostrogylus（20.5%），Haemonchus（10.6%），Strongyloides（51.5%）in all groups.The age-specific intensity for Eimeria was in following order:kids（92.6%），adults（86.6%）and yearlings（51.5%）and the difference was not significant（P＞0.05）.The concurrent infections between GINs and Eimeria were 67.94%in all groups.Polyparasitisms of Trichostrongylus（17.4%），Haemonchus（15.15%）and Strongyloides（40.4%）with Eimeria had infected most adult goats.Dual infections of Trichostrongylus with Eimeria and Trichuris with Eimeria were significantly higher in adults than year lings and kids（P＜0.05）.

Conclusions:GINs and Eimeria were widely distributed in the goats leading to higher risk of morbidity and mortality.

1.Introduction

In recent years，the global goat population has exhibited a rapid growth over 875.5 million heads of which Asia is observed to have the largest goat population［1］mostly in developing countries including China，India，Malaysia and Pakistan［2，3］. As a result of this huge expansion，the growth rate of goat population in Malaysia has also consistently increasing from 2010 to 2013 with a total of 482280 goats nationwide［4］. Malaysians' preferences for goatmeat have gradually increasing as an alternative to other red meats［5］due to its low total and saturated fat compared to beef，pork，lamb and chicken［6，7］.Year by year，the levels of mutton production in Malaysia are estimated to increase from 8.99%to 35.00% with the increasing of the goat population to 1.5 million heads［5］.

However，gastrointestinal nematodes（GINs）have becoming an important helminth group that causes direct damages to the livestock.They remain as one of the main constraints to ruminant productions especially goats，since they can cause several

subclinical effects such as hypoproteinemia，growth depression，reduction in milk yield，loss of appetite and digestive inefficiency［6，7］.In Malaysia，gastrointestinal nematodiasis，referring mainly to haemonchosis was classified as one of the most important causes of mortality and morbidity in small ruminants［8］.In the past two decades，a study conducted in seven farms in Penang Island［9］had revealed Haemonchus contortus（H.contortus）as the most prevalent nematode species in fected the studied goats.Years afterwards，Ch and rawathani et al.［10］also reported that H.contortus had predominated the goats in Peninsular Malaysia.Besides that，there were reports from many countries world wide on prevalence of GINs including H.contortus，Oesophagostomum spp.，Cooperiaspp.and Trichostrongylus spp.，which had caused massive decline in milk and meat productions leading to severe economic losses in the goat industry［11-14］.

Another important parasitic infections in small ruminants that prevail widely in many parts of the world either clinically or subclinically is coccidiosis.It is caused by coccidian parasites of the genus Eimeria which contribute to enteric disease especially in young or stressed goats under poor farm management that leads to high mortality in goat kids［15］.Other than that，studies in many countries such as Czech Republic［16］，Jordan［17］，Malaysia［18］，Poland［19］and Sri Lanka［20］have indicated that goat coccidiosis has a great economic importance to the livestock productions due to the clinical disease（diarrhea）and sub-clinical disease（poor weight gain）caused by the parasite especially goats kept in large numbers under various management systems.Furthermore，GINs and coccidia infections among goats are also closely associated with other several factors including poor farm management，ingestion of contaminated food and water，nutritional deficiencies，age of the goats and also climate factor［21］.

In the previous year，40 goats from two farms in Kuala Terengganu，Terengganu were examined for endo-and ectoparasites.This study only compared the prevalence and intensity of parasitic infections between the two farms［22］.Unlike the present study，it evaluated the prevalence of coccidian and GINs associated with different age groups in three different farms from three different districts of Terengganu（Kuala Terengganu，Setiu and Besut）.More importantly，there is a no established data available on the prevalence of concurrent GINs and Eimeriaspp.infections in Terengganu with relation to the three different age groups of goats.Thus，the present study aimed to determine the prevalence of gastrointestinal nematodiasis and coccidiosis from three age groups of goats and provided information on the predominance of concurrent GINs and Eimeriaspp.in goats from three farmsin Terengganu.The results obtained should provide a foundation for the improved control of both GINs and coccidia infections in goats.

2.Materials and methods

2.1.Ethics statement

The present study had been approved by the Department of Veterinary Services，Terengganu dated 18th May 2014（Ref. JPV.TR 03/03/13）.All experimental procedures involving animals were conducted in accordance to Department of Veterinary Services，Terengganu and approved by the Department of Veterinary Services，Terengganu.

2.2.Study areas and subjects

This study was conducted in three commercialized farms in Terengganu（a）Farm 1:Kuala Terengganu，（b）Farm 2:Setiu and（c）Farm 3:Besut.The farms were rearing more than 100 goats mainly for milk and meat productions.A total of 287 goats including 81 goat kids，74 year lings and 132 adult goats were sampled once between February and April 2015.The weather across these three districts were uniformly warm and rainy with average rainfall ranging between 200 mm and 400 mm.The animals were catergorised according to guidelines provided by the Department of Veterinary Services，Terengganu［（a）kids: age below 12 months，（b）yearlings:12 months but under 24 months old，（c）adults:above 24 months old］.The kids had received colostrum for the first 2-3 days of life and kept together with their dams only in the morning whereas most year lings and adult goats were turned out for grazing during the day and penned separately according to age group at night.

2.3.Faecal sample collection

Faecal samples were collected directly from the rectum of randomly selected animals using sterile disposable plastic gloves and placed in a faecal container.For each animal，the date of sampling，age，animal identification number and sex were recorded.The samples were transported on the same day of each sample collection in an airtight box cooler with dry ice packs to the Laboratory of Parasitology，Integrated Centre for Animal Care and Use，International Islamic University Malaysia and then kept at 4°C for a maximum of 48 h before analysis.

2.4.Examination of faecal samples

Three grams of each faecal sample were mixed with 45 mL of floatation fluid，sodium chloride（1.2 specific gravity）to recover the intestinal eggs and oocysts from the faecal debris［23］.The presence of parasite stages（eggs or larvae of helminths，oocysts of coccidian）was identified on based on their morphological characteristics［24，25］using10×and40× magnification.

2.5.Statistical analysis

The overall prevalence was calculated as a percentage of number of animals infected in the total number of animals examined.Relative prevalence for different species of parasites，respectively，was calculated and stratified by three age groups（kids，yearlings and adults）.In all analyses，confidence level was held at 95%and P value of≤0.05 was set as the level of significance.The analysis was performed using SPSS software for Windows version 22.0（SPSS Inc.，Chicago，USA）.

3.Results

A total of 287 goats from three farms in Besut，Setiu and Kuala Terengganu were successfully examined.The present study revealed that the overall prevalence of GINs in all three age groups was 77.7%（223/287）while 89.2%（256/287）goats examined were found to be infected with coccidian（Table 1）. The prevalence of coccidia was significantly higher than GINs（χ2；P＜0.05）.

Table 1　Overall prevalence of GINs and coccidia infections in kid，yearling and adult goats［n（%）］.

As summarized in Table 1，the highest prevalence of GINs infection was observed in yearlings（87.8%），followed by the adults（86.4%）and kids（53.1%）.The association between the prevalence of GIN infections and the age groups was identified to be significant（χ2；P＜0.05）.Generally，coccidia oocysts were detected in 256 of 287 faecal samples（89.2%），including 75 samples originating from goat kids（92.6%），68 samples from yearlings（91.9%）and 113 samples from adult goats（85.6%）. Contradicting to nematode infections，there was no significant association between the prevalence of coccidial infection and the age groups（χ2；P＞0.05）.

A total of four genera of GINs were found in the studied goat including Trichostrogylus spp.，Haemonchus spp.，Strongyloides spp.and Trichurisspp（Table 2）.The most prevalent GINs observed was Strongyloides spp.（45.6%）followed by Trichostrogylus spp.（14.3%），Haemonchus spp.（9.0%）and Trichurisspp.（8.7%）.Coccidia oocysts belonging to the genus of Eimeria spp.（89.2%）was the only coccidian parasite identified in this study.

Table 2　Prevalence of different species of GIN and coccidia in three age groups［n（%）］.

The adult goats were observed to have the highest infections of Trichostrogylus spp.，Haemonchus spp.，Strongyloides spp. among all three age groups of goats.However，only the prevalence of Trichostrogylus spp.was significantly higher in adult goats than in year lings and kids（χ2；P＜0.05）.The prevalence of Eimeriaspp.oocysts in kid，yearling and adult goats were 92.6%，91.9%and 85.6%，respectively，and the difference was not significant（χ2；P＞0.05）.

Table 3 demonstrates the concurrent Eimeria spp.and four species of GINs infections（Trichostrongylus spp.，Haemonchus spp.，Strongyloides spp.and Trichurisspp.）in the studied goats. Mixed infections were generally present in all age groups but the frequency of multiple infections varied substantially among those groups.Overall prevalence for mixed infection rate of nematodes and Eimeriaspp.was 68.29%（196/287）whereby cases of co-infection between Strongyloides spp.and Eimeria spp.（38.33%；110/287）were the highest among the mixed infections.

Table 3　Concurrent Eimeria spp.and GINs.

Among the dual infections in three age groups，pairs of Trichostrongylus spp.，Haemonchus spp.and Strongyloides spp. with Eimeria spp.had predominated the adult goats with prevalence of 17.42%，15.15%and 40.91%，respectively.Meanwhile，polyparasitism of Trichurisspp.with Eimeriaspp.had infected most in the year lings than in kids and adults with percentage of 18.92%.Statistically，there were significant differences between concurrent infections of Trichostrongylusspp.，Trichurisspp.with Eimeriaspp.with different age groups（χ2；P＜0.05）.

4.Discussion

In Malaysia，the detection of GINs in cattle and sheep have been widely reported.However，there is a conspicuous lack of relevant data focusing on gastrointestinal nematodiasis isolated from goats in Malaysia.Current finding showed that 77.7%of studied animals were positive with GINs，predominated by Strongyloidesspp.（45.6%）and least predominant GINs was Trichurisspp.（8.7%）.The finding was closely related to the result reported by Hassan et al.［26］in Bangladesh who found 51.74%of goats were in fected with Strongyloidesspp. However，this result differs from studies done in Penang Island［9］and Selangor［27］，Malaysia that revealed the most prevalent nematode species infected the goats was Haemonchusspp.with percent age of45.7%，46.3%，respectively.Likewise，the predominance of Haemonchusspp. in goats has also been reported in Nigeria（87.3%）［28］，South Africa（54%）［21］and Sri Lanka（90%）［20］.On the other hand，the result of low occurrence of Trichurisspp.in present study corroborates the observations of goats in Nigeria（2.43%）［28］. The high infection rate of GINs might due to goat grazing activities on pastures contaminated with third stage infective larvae of parasitic nematodes［29］.The goats were allowed to graze for long hours（4-6 h）in the grazing land，thereby increasing the exposure to infections through ingestion of larvae that freely lived in the grazing land［30］.Additionally，sufficient humidity and favor able temperature，create an environment that enhances the development of GINs and sustains the survival of the infective stage of larvae，leading to high pasture contamination［19］.

In the present study，the overall prevalence of coccidian oocysts in goats was 89.2%.The finding was in agreement with reports made in different states of Malaysia（Selangor，Terengganu，Perak and Pahang）［18，22，29，31］whereby the infection rates were 85%（n=81），92.57%（n=162），89%（n=815）and 100%（n=40），respectively.On the contrary，a study conducted by Chandra wathani et al.［32］in Perak has revealeda lower total prevalence of coccidiosis with percentage of 48.77%（n=203）.Similar to our findings，a relatively high prevalence of coccidiosis from various studies conducted around the globe was reported.The frequency rates were 83.43%（n=350）in Iran［33］，98.6%（n=144）in Portugal［34］，97%（n=277）in the USA［35］，80.35%（n=173）in Western Poland［19］，87.94%（n=175）in China［36］，but in Southern Botswana［37］，India［38］and Pakistan［39］，13.3%（n=143），42.86%（n=144）and 43.87%（n=310）goats were positive for coccidian infection，respectively，indicating a lower prevalence of infection compared to present study.

The prevalence coccidial oocysts from genus of Eimeria was the highest in kids（92.6%）compared to year lings（91.9%）and adult goats（85.6%）which were consistent with the previous observations［34，36，37］.This has been attributed to underdeveloped and lower immunological resistance towards coccidian infection in goat kids compared to older goats［36，38］. Nevertheless，the well-developed resistance observed in adult goats appeared to be relative rather than absolute since adults continued to harbor Eimeriaspp.oocytes in faeces，which contributed as source of infection for younger goats［35］.

We noted that 68.29%（196/287）studied animals had concurrent infections of Eimeriaspp.and GINs in this study.In comparison with previous studies，the prevalence of concurrent infections was much lower than Sri Lanka（78%；n=203）［20］but higher than India（17.22%；n=273）［38］and Korea（29.3%；n=241）［40］.Additionally，concurrent infections in all three age groups were observed in this study complementary to reports by Radfar et al.［41］and Faizal and Rajapakse［20］. Polyparasitisms in this study perhaps are due to repeated exposure to infections of both GINs and Eimeriaspp.［20］.

In conclusion，the present study indicated that both GINs and Eimeriaspp.as single or concurrent infections are prevalent in goats of these three farms in Terengganu.The parasitic loads pose a great economic impact and constraint to the goat industry in Terengganu in achieving better and sustainable productions. Therefore，these findings suggest for a need of well-coordinated，sanitary monitoring of goat farms by field veterinarians and dissemination of knowledge on GINs and Eimeriaspp.to animal handlers and farmers to minimize the occurrence of infections.

Conflict of interest statement

We declare that we have no conflict of interest.

Acknowledgments

The authors would like to express our gratitude to the owners of the farms and logistic support by the Department of Veterinary Services，Terengganu.The study was funded by the Ministry of Higher Education，Malaysia under the grant of Research Acculturation Grant Scheme（RAGS）（Grant No.13-010-0073）.

References

［1］Food and Agriculture Organization of the United Nations.FAO Statistical Yearbook 2013.World food and agriculture.Rome: Food and Agriculture Organization of the United Nations；2013.［Online］Available from:http://www.fao.org/docrep/018/i3107e/ i3107e00.htm［Accessed on 15th June，2015］

［2］Aziz MA.Present status of the world goat populations and their productivity.Lohmaan Inf 2010；45:42-52.

［3］Food and Agriculture Organization of the United Nations.FAO statistical yearbook 2007-2008.Rome:Food and Agriculture Organization of the United Nations；2008.［Online］Available from: http://www.fao.org/economic/ess/ess-publications/ess-yearbook/ fao-statistical-yearbook-2007-2008/b-agricultural-production/en/［Accessed on 10th July，2015］

［4］The Government of Botswana.Department of Veterinary Services. Livestock statistics.Gaborone:The Government of Botswana；2009.［Online］Available from:http://www.gov.bw/en/Ministries-Authorities/Ministries/MinistryofAgriculture-MOA/Departmentsof-MOA/Department-of-Veterinary-Services-DVS/［Accessed on 17th May，2015］

［5］Kaur B.Consumer preference for goat meat in Malaysia:market opportunities and potential.J Agribus Mark 2010；3:40-55.

［6］Luginbuhl JM.Meat goat production in North Carolina.North Carolina:North Carolina State University；2015.［Online］Available from:http://citeseerx.ist.psu.edu/viewdoc/download？doi=10. 1.1.464.5160&rep=rep1&type=pdf［Accessed on 15th May，2015］

［7］Gebhardt SE，Thomas RG.Nutritive value of foods.Maryland: United States Department of Agriculture；2002.［Online］Available from:https://www.ars.usda.gov/SP2UserFiles/Place/80400525/Data/ hg72/hg72_2002.pdf［Accessed on 20th May，2015］

［8］Nor-Azlina AA，Sani RA，Ariff OM.Management practices affecting helminthiasis in goats.Pertanika J Trop Agric Sci 2011；34:295-301.

［9］Wahab AR，Adanan CR.Observations on the worm egg counts and their nematode species in goats from the North-East District of Penang Island，Peninsular Malaysia.Pertanika 1992；15:221-4.

［10］Chandrawathani P，Adnan M，Waller PJ.Antihelminthic resistance in sheep and goats farms on Peninsular Malaysia.Vet Parasitol 1999；82:305-10.

［11］Ibrahim N，Tefera M，Bekele M，Alemu S.Prevalence of gastrointestinal parasites of small ruminants in and around Jimma Town，Western Ethiopia.Acta Parasitol Glob 2014；5:26-32.

［12］Ma J，He SW，Li H，Guo QC，Pan WW，Wang XJ，et al.First survey of helminths in adult goats in Hunan Province，China.Trop Biomed 2014；31:261-9.

［13］Raza MA，Younas M，Schlecht E.Prevalence of gastrointestinal helminths in pastoral sheep and goat flocks in the Cholistan Desert of Pakistan.J Anim Plant Sci 2014；24:127-34.

［14］Zanzani SA，Gazzonis AL，Di Cerbo A，Varady M，Manfredi MT. Gastrointestinal nematodes of dairy goats，antihelmithic resistance and practices of parasite control in Northern Italy.BMC Vet Res 2014；10:114.

［15］Ratanapob N，Arunvipas P，Kasemsuwan S，Phimpraphai W，Panneum S.Prevalence and risk factors for intestinal parasite infection in goats raised in Nakhom Pathom Province，Thailand. Trop Anim Health Prod 2012；44:741-5.

［16］Koudela B，Bokov′a A.Coccidiosis in goats in the Czech Republic. Vet Parasitol 1998；76:261-7.

［17］Abo-Shehada MN，Abo-Farieha HA.Prevalence of Eimeria species among goats in Northern Jordan.Small Rumin Res 2003；49:109-13.［18］Jalila A，Dorny P，Sani R，Salim NB，Vercruysse J.Coccidial infections of goats in Selangor，Peninsular Malaysia.Vet Parasitol 1998；74:165-72.

［19］Balicka-Ramisz A.Studies on coccidiosis in goats in Poland.Vet Parasitol 1999；81:347-9.

［20］Faizal AC，Rajapakse RP.Prevalence of coccidia and gastrointestinal nematode infections in cross bred goats in the dry areas of Sri Lanka.Small Rumin Res 2001；40:233-8.

［21］Gwaze FR，Chimonyoa M，Dzamab K.Prevalence and loads of gastrointestinal parasites of goats in the communal areas of the Eastern Cape Province of South Africa.Small Rumin Res 2009；84:132-4.

［22］Khadijah S，Andy TFH，Khadijah SSAK，Khairi AKM，Nur Aida H，Wahab AR.Parasite infection in two goat farms located in Kuala Terengganu，Peninsular Malaysia.Asian J Agric Food Sci 2014；2:463-8.

［23］Dryden MW，Payne PA，Ridley R，Smith V.Comparison of common fecal floatation techniques for the recovery of parasite eggs and oocysts.Vet Ther 2005；6:15-28.

［24］Urquhart GM，Armour J，Duncan JL，Dunn AM，Jennings FW. Veterinary parasitology.2nd ed.Glasgow:Wiley-Blackwell；1996，p.3-137.

［25］van Houtert MF，Sykes AR.Implications of nutrition for the ability of ruminants to withstand gastrointestinal nematode infections.Int J Parasitol 1996；26:1151-67.

［26］Hassan MM，Hoque MA，Islam SKMA，Khan SA，Roy K，Banu Q. A prevalence of parasites in black Bengal goats in Chittagong，Bangladesh.Int J Livest Prod 2011；2:40-4.

［27］Dorny P，Symoens C，Jalila A，Vercruysse J，Sani R.Strongyle infections in sheep and goats under the traditional husbandry system in Peninsular Malaysia.Vet Parasitol 1995；56:121-36.

［28］Nwigwe JO，Njoku OO，Odikamnoro OO，Uhuo AC.Comparative studies of intestinal helminthes and protozoa of cattle and goats in Abakaliki metropolis of Ebonyi State，Nigeria.Adv Appl Sci Res 2013；4:223-7.

［29］ZainalabidinFA，RaimyN，YaacobMH，MusbahA，Bathmanaban P，Ismail EA，et al.The prevalence of parasitic infestation of small ruminant farms in Perak，Malaysia.Trop Life Sci Res 2015；26:1-8.

［30］Sutherland I，Scott I.Gastrointestinal nematodes of sheep and cattle:biology and control.London:John Wiley and Sons；2009，p.61-75.

［31］Tahir MS，Ibrahim TAT，Babjee SMA.Study on coccidia infection and species in Cyprus Shami goat population.In:The 6th proceedings of the Seminar on Veterinary Sciences；2011 Jan 11-14；Kuala Lumpur，Malaysia.p.46-50.

［32］Chandrawathani P，Nurulaini R，Adnan M，Premalaatha B，Khadijah S，Jamnah O，et al.A survey of parasitic infection on small ruminant farms in Kinta and Hilir Perak Districts，Perak，Malaysia.Trop Biomed 2009；26:11-5.

［33］Hashemnia M，Rezaei F，Chalechale A.Prevalence，intensity，and pathological lesions of Eimeria infection in goats in Western Iran. Comp Clin Pathol 2015；24:805-10.

［34］Silva LM，Vila-Vi?osa MJ，Nunes T，Taubert A，Hermosilla C，Cortes HC.Eimeria infections in goats in Southern Portugal.Rev Bras Parasitol Vet 2014；23:280-6.

［35］Kahan TB，Greiner EC.Coccidiosis of goats in Florida，USA. Open J Vet Med 2013；3:209-12.

［36］Wang CR，Xiao JY，Chen AH，Chen J，Wang Y，Gao JF，et al. Prevalence of coccidial infection in sheep and goats in Northeastern China.Vet Parasitol 2010；174:213-7.

［37］Sharma S，Busang M.Prevalence of some gastrointestinal parasites of ruminants in southern Botswana.Bots J Agric Appl Sci 2013；9: 97-103.

［38］Sharma DK，Agrawal N，Mandal A，Nigam P，Bhushan S.Coccidia and gastrointestinal nematode infections in semi-intensively managed Jakhrana goats of semi-arid region of India.Trop Subtrop Agrosyst 2009；11:135-9.

［39］Rehman TU，Khan MN，Khan IA，Ahamad M.Epidemiology and economic benefits of treating goat coccidiosis.Pak Vet J 2011；31: 227-30.

［40］Gebeyehu EB，Seo MG，Jung BY，Byun JW，Oem JK，Kim HY，et al.Prevalence of gastrointestinal parasites in Korean Native goats（Capra hircus aegagrus）.J Anim Plant Sci 2013；23:986-9.

［41］Radfar MH，Sakhaee E，Bafti MS，Mohammadi HH.Study on gastrointestinal parasitic infections of Raeini goats.Iran J Vet Res 2011；12:76-80.

14 Mar 2016

in revised form 8 Apr 2016

Afzan Mat Yusof，Assistant Professor，Ph.D，Department of Biomedical Science，Kulliyyah of Allied Health Sciences，International Islamic University Malaysia，Jalan Sultan Ahmad Shah，Bandar Indera Mahkota，Kuantan 25200，Pahang，Malaysia.

Tel:+60129656253

E-mail:afzan@iium.edu.my

All experimental procedures involving animals were conducted in accordance to Department of Veterinary Services（DVS），Terengganu and approved by the Department of Veterinary Services（DVS），Terengganu.

Foundation Project:Supported by Ministry of Higher Education，Malaysia under the grant of Research Acculturation Grant Scheme（RAGS）（Grant No.13-010-0073）.

Peer review under responsibility of Hainan Medical University.The journal implements double-blind peer review practiced by specially invited international editorial board members.

2221-1691/Copyright?2016 Hainan Medical University.Production and hosting by Elsevier B.V.This is an open access article under the CC BY-NC-ND license（http:// creative commons.org/licenses/by-nc-nd/4.0/）.