Sains Malaysiana 51(6)(2022): 1677-1686

http://doi.org/10.17576/jsm-2022-5106-06

Isolation and Establishment of eimeria tenella Populations from Local Broiler Chicken Farms

(Pemencilan dan Penghasilan Populasi Eimeria tenella daripada Ladang Ayam Pedaging Tempatan)

SHU-SAN LOO^1,2, MAZLAN MOHAMED³, FARAH SHAFAWATI MOHD-TAIB¹, CHOON-KIAT KHOO⁴ & KIEW-LIAN WAN^1,*

¹Department of Biological Sciences and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor Darul Ehsan, Malaysia

²Agro-Biotechnology Institute, National Institutes of Biotechnology Malaysia, c/o MARDI Headquarters, 43400 Serdang, Selangor Darul Ehsan, Malaysia

³One Biotech Sdn. Bhd., No. 9-1, Jalan 5/11C, Seksyen 5, Bandar Baru Bangi, 43650 Bangi, Selangor Darul Ehsan, Malaysia

⁴Veterinary Research Institute, Department of Veterinary Services Malaysia, No. 59, Jalan Sultan Azlan Shah, 31400 Ipoh, Perak Darul Ridzuan, Malaysia

Received: 20 March 2021/Accepted: 29 November 2021

Abstract

Coccidiosis is a major intestinal disease in the chicken production industry that is caused by one or more of the seven Eimeria species known to infect chickens. Development of effective diagnostic methods and controls requires comprehensive knowledge on the Eimeria species in the local population, particularly the pathogenic E. tenella. In this study, 35 faecal samples were collected from local chicken farms, and through microscopic observation, nine samples (26%) were found to contain Eimeria oocysts. Subsequently, two positive samples, namely NSN6 and SGR6, were selected and propagated via passage in coccidian-free chickens. Species identification analyses based on oocyst morphological characterisation suggested the presence of small-sized oocysts (E. acervulina and/or E. mitis) and medium-sized oocysts (E. necatrix, E. tenella, and/or E. praecox) in NSN6 while in SGR6, the large-sized oocysts (E. brunetti and/or E. maxima) were also present in addition to the small and medium-sized oocysts. Subsequently, species identification with PCR using species-specific primers was successful in determining the presence of specific Eimeria species, which are E. acervulina and E. tenella in NSN6, and E. acervulina, E. tenella, and E. maxima in SGR6. The E. tenella populations of NSN6 and SGR6, namely EtNSN6 and EtSGR6, respectively, were established via passage in coccidian-free chickens and oocyst recovery from the caeca. The purity of EtNSN6 and EtSGR6 populations were confirmed based on oocyst morphological characterisation and PCR. The established EtNSN6 and EtSGR6 populations would be useful as research models for local strains in the development of more effective control methods.

Keywords: Coccidiosis; local strains; oocyst morphology

Abstrak

Koksidiosis merupakan penyakit saluran usus utama dalam industri penternakan ayam yang disebabkan oleh satu atau lebih daripada tujuh spesies Eimeria yang diketahui menjangkiti ayam. Pembangunan kaedah diagnostik dan kawalan yang berkesan memerlukan pengetahuan tentang spesies Eimeria dalam populasi tempatan, khususnya E. tenella yang patogenik. Dalam kajian ini, sejumlah 35 sampel tinja ayam diperoleh daripada ladang ayam tempatan dan melalui pemerhatian mikroskop, didapati sembilan sampel (26%) mengandungi oosista Eimeria. Seterusnya, dua sampel positif, iaitu NSN6 dan SGR6, dipilih dan dibiakkan secara pasaj dalam ayam bebas koksidia. Analisis pengenalpastian spesies berdasarkan pencirian morfologi oosista mencadangkan kehadiran oosista yang bersaiz kecil (E. acervulina dan/atau E. mitis) dan oosista yang bersaiz sederhana (E. necatrix, E. tenella dan/atau Eimeria praecox) dalam NSN6 manakala dalam SGR6, oosista yang bersaiz besar (E. brunetti dan/atau E. maxima) juga hadir di samping oosista yang bersaiz kecil dan sederhana. Selanjutnya, pengenalpastian spesies dengan PCR menggunakan pencetus khusus terhadap spesies telah berjaya mengenal pasti kehadiran spesies Eimeria secara khusus, iaitu E. acervulina dan E. tenella dalam NSN6 serta E. acervulina, E. tenella dan E. maxima dalam SGR6. Populasi E. tenella NSN6 dan SGR6, iaitu masing-masing EtNSN6 serta EtSGR6 dihasilkan melalui pasaj dalam ayam bebas koksidia dan perolehan oosista daripada bahagian sekum. Ketulenan populasi EtNSN6 dan EtSGR6 kemudian disahkan secara pencirian morfologi oosista dan PCR. Populasi EtNSN6 dan EtSGR6 adalah berguna sebagai model penyelidikan untuk strain tempatan dalam pembangunan kaedah pengawalan yang lebih berkesan.

Kata kunci: Koksidiosis; morfologi oosista; strain tempatan

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^*Corresponding author; email: klwan@ukm.edu.my