Blood Parasites of Bali Ducks Sampled from Traditional Farming System in Bali.

IDA AYU PASTI APSARI1) , , IWAN HARYONO UTAMA1), NYOMAN SUARSANA1), ANAK AGUNG AYU MIRAH ADI1), IDA BAGUS OKA WINAYA1), I GUSTI MADE KRISNA ERAWAN1), AND YOSHIHIRO HAYASHI2).

1) Faculty of Veterinary Medicine, Udayana University-Kampus Bukit Jimbaran-Bali. E-mail : [email protected]
2) Faculty of Agriculture and Life Sciences, Tokyo University-Tokyo Japan; e-mail : [email protected]

SUMMARY
Research to confirm blood parasites infection in Bali ducks based on blood smear examination has been conducted. In this research, we took 80 ducks as a sample from some areas in Bali. A thin blood smear had been made and stained with Giemsa. Data were collected and tabulated by means of descriptive method. Results : 54 of ducks (67.5%) infected with blood parasites such as : Plasmodium sp. which infected 44 of 80 (55%) ducks; Leucocytozoon sp.infected 19 of 80 (23.75%) ducks and 9 ducks (11.25%) were infected by both of those parasites. Based from the origin of ducks, the highest incidence of Plasmodium sp. infection is in regency of Buleleng (80%), and the highest incidence of Leucocytozoon sp. infection is in Badung regency (50%). According to sex preference, 24 of 30 male ducks (80%) and 30 of 50 female ducks (60%) were infected. There was no significant difference in prevalence of infection between male and female ducks (P>0,05).

Keywords : Bali ducks, blood parasites
RINGKASAN
Penelitian ini bertujuan mengetahui infeksi parasit darah pada itik Bali berdasarkan pemeriksaan sediaan ulas darah. Penelitian ini menggunakan 80 ekor itik yang terdiri dari 30 ekor itik jantan dan sisanya betina, itik diambil dari beberapa daerah di Bali. Sediaan ulas darah dibuat sediaan ulas darah tipis dan diwarnai dengan Giemsa. Data dikumpulkan serta ditabulasi secara diskriptif. Hasil : sebanyak 54 ekor itik (67.5%) positif terinfeksi parasit darah seperti Plasmodium sp. yang menginfeksi 44 ekor (55%); Leucocytozoon sp. menginfeksi 19 ekor (23.75%); dan sisanya sebanyak 9 ekor (11.25%) terinfeksi oleh kedua jenis parasit tersebut. Berdasarkan daerah asal usul itik, infeksi Plasmodium sp. yang tertinggi terdapat di kabupaten Buleleng (80%), sedangkan kejadian infeksi Leucocytozoon sp. yang tertinggi terdapat di Kabupaten Badung (50%). Berdasarkan jenis kelamin, sebanyak 24 dari 30 ekor itik jantan (80%) serta 30 dari 50 ekor itik betina (60%) terinfeksi parasit darah. Tidak ada perbedaan prevalensi infeksi antara itik jantan dan betina (P>0,05).

Kata kunci : Itik Bali, parasit darah.

Introduction
Bali is an island located in Indonesia, besides having beautiful sceneries and full of harmony in their life and culture, it also has some specific animals such as: Bali cattle, Bali dogs, Bali starlings, and of course Bali ducks. Generally, the local community keeps small flock of ducks as a secondary activity to their main agricultural activities and they are still reared in a traditional way (Figure 1). So far they are kept not only for income generation, but also for ceremonial or religious purposes. Bali ducks is an ancient breed which originator of Indian Runner. Compared with general morphology of common ducks and Domestic Waterfowl Club, Bali ducks are classified as light ducks (www.domestic-waterfowl.co.uk/page3.htm-37k). They are also known as penguin ducks with their slim body and they stands upright (Rasyaf, 1982), according to Baldassare and Bollen (1994), ducks in Indonesia generally belong to Anas poecilorrhyncha (spot-billed duck). Bali ducks have two-major phenotypes, the sumi-feathered ducks (sumi means: color looks like rice straw, brownish) and the white feathered ducks (Matram, 1984). Sometimes we can find black color (sikep) ducks. General inspection shows that there are no other morphologically significant differences except their feather’s color

Figure 1. Ducks are traditionally farmed, just for saving

Information about some aspects related to avian health and diseases in Bali (including in Bali ducks) is still little and not yet established so far. Data from Bali Provincial Livestock Services showed that the majority of health problems in ducks and chicken in Bali is avian pasteurelosis, pullorum, avian coryza, Newcastle disease, leucocytozoonosis, and intestinal protozoa such as Eimeria sp. Non specific diarrhea are still a problem in some areas of Indonesia, including Bali (Farm Animal Data, Bali’s Provincial Livestock Services, 2002).
So, in this research We will conduct blood parasites examination in Bali ducks (Jain, 1986; Qureshi, 1988).

MATERIALS AND METHODS
Materials used are: object glass and cover glass, microscope (Meiji, Japan), balances (Navigator-O-Haus, Germany), electronic pH meter (Hanna Instruments, HI-8519, USA). Chemicals used are : aquadest, absolute methanol, alcohol 70%, cotton, Giemsa solution, immersion oil, and xylol (all produced by Merck, USA). Giemsa solution was made by mixing of one ml of stock solution with nine milliliters of aquadest.

Figure 2. Map of 8 regencies as the places of taking duck samples including the
Municipality of Denpasar

This research was performed by taking samples in some areas listed in Figure 2 above, all 80 productive ducks caught (aged 8 – 10 month) from 8 districts in Bali except Negara were examined. A drop of blood from wing-vein was smeared on glass object, blood smear was fixed in absolute methanol for 5 minutes and stained with Giemsa for 30 minutes. After finished, wash the stained slide with distilled water and dried (Coles, 1980; Houwen, 2000). Blood smear examination was focused on blood parasites (Demsey and Kennedy, 1999; Fludge, 1997; Jones, 1999; Phillips, 1999).

Data analysis
Health data were tabulated and analyzed by means of descriptive methods and c2 test for contingency table is performed to analyse between infection status and sex of ducks (Steel and Torrie, 1991).

RESULTS AND DISCUSSION

Table 1. Some parasites found in Bali ducks, and their incidence in general.
Parasites % Incidence
Leucocytozoon sp. 23,75% (19 of 80)
Plasmodium sp. 55% (44 of 80)
Both Leucocytozoon sp. and Plasmodium sp. 11.25% (9 of 80)

Table 2. Distribution of blood parasites according to some area surveyed in Bali.
Regencies Total Parasites Plasmodium sp. Leucocytozoon sp.
+ -
Badung 12 7 5 16,7% (2 of 12) 50% (6 of 12)
Tabanan 10 7 3 50% (5 of 10) 20% (2 of 10)
Denpasar municipal 8 5 3 62,5% (5 of 8) 12,5% (1 of 8)
Klungkung 10 7 3 60% (6 of 10) 20% (2 of 10)
Gianyar 10 7 3 60% (6 of 10) 40% (4 of 10)
Buleleng 10 8 2 80% (8 of 10) -
Karangasem 10 7 3 70% (7 of 10) 10% (1 of 10)
Bangli 10 6 4 60% (6 of 10) 30% (3 of 10)
Total 80 54 26
Note : - not detected

Table 3. Distribution of blood parasites according to sex of ducks
Parasites Male Female Total
+ 24 30 54
- 6 20 26
Total 30 50 80
Note : P>0,05

Figure 3 : Trophozoid of Plasmodium sp. in uck’s erythrocyte (objective 100x) Figure 4. Gamon of Leucocytozoon sp.

The existence of protozoa both in blood can be seen in Table 1., Table 2 shows kind of parasites found related to regencies surveyed in Bali, and Table 3 shows distribution of parasites according to sex of ducks. From the blood smear examination, 67,5% (54 of 80 ducks) were infected by blood parasites. Plasmodium sp. is the most prevalence (55%), followed by Leucocytozoon sp. and mixed infections caused by both agents. Apsari et al. (1999) showed more than 53,58% chicken in Bali were infected by Leucocytozoon sp. Winarsih and Widhyari (1998) reported both Plasmodium sp. and Leucocytozoon sp. as infectious agent in Kedu chicken.
Leucocytozoon sp. is a common causative agents in several duck’s species such as Anas platyrhyncos (Clark, 1980), Aix sponsa (Dell and Robbins, 1994), Anas acuta (Williams, et al. 1977). Among those ducks, infection caused by Leucocytozoon sp. is the highest prevalence. Plasmodium sp. infection also occurred in Anas rubipes and Anas carolinensis (Bennet et al. 1975), Aix sponsa (Thul et al. 1980), Anas discors, Aythya affinis, and Aythya collaris (Loven et al. 1980), even raptors (Buteo jamaicensis, Haliaeetus leucocephalus, and Otus asio) could be infected by Plasmodium elongatum (Nayar, et al. 1998). From those data, Leucocytozoon sp. and Plasmodium sp. could infect many avian hosts.
In Bali, L. sabrasezi is the most dominant agent (99,48%) causing leucocytozoonosis in kampong chicken, rest of those chicken were infected by L. cauleryi (Gunawan, et al. 1984). Our epidemiological data shows Plasmodium sp. infection is the highest prevalence, more than fifty percent infected ducks originated from regencies in Bali (except Badung Regency). It is interesting phenomenon, because Culex sp. is vectors of those parasites found in many wet places in Denpasar such as water trenches and drains. Anopheles sp. is also vectors found in many rice fields (Aryani, 2003). This is also a habitat of bali ducks.
From Table 3 we could see that is not significant difference between sex and parasite infection (P>0,05). This result is supported by Apsari (1999).
In Indonesia, especially Bali, knowing that ducks are raised together with chicken, it is strongly assumed that there is cross infection by the agents of the cause of avian leucocytozoonosis and plasmodiosis. Darmono et al. (1982) stated traditionally farmed chicken were sensitive to blood parasite infection, it could be same for ducks traditionally farmed. It need more comprehensive study such as comparison of isolates from chicken and ducks.

CONCLUSION
Even those ducks were clinically healthy, 67,5% of ducks examined were infected by blood protozoan parasites consisted by Plasmodium sp. (55%), Leucocytozoon sp. (23,75%), both Plasmodium sp. and Leucocytozoon sp. (11,25%). According to sex, 80% male and 60% female ducks were infected. The highest incidence of Plasmodium sp. infection is in regencies of Buleleng (80%), Karangasem (70%), Denpasar (62,5%), Bangli, Gianyar, and Klungkung (60%) respectively and the highest incidence of Leucocytozoon sp. infection is in Badung regency (50%).

Acnowledgement
This study was part of the collaborative research between Japan Society for the Promotion of Science (JSPS) and Faculty of Veterinary Medicine Udayana University, Bali-Indonesia entitled : Determination of Health Status of Bali Ducks for Sustainable Agriculture in Bali. It was funded by JSPS. We also thank Prof. Yoshihiro Hayashi, DVM., PhD. From Tokyo University, for being counterpart in this study.

REFERENCES :

Apsari, I. A. P., I. B. N. Swacita and N. A. Suratma. 1999. Prevalensi infeksi Leucocytozoon sp. pada ayam buras serta status hematologinya. Research report Faculty of Veterinary medicine, Udayana University.

Apsari, I. A. P. 1999. Pengaruh kondisi wilayah asal dan jenis kelamin terhadap prevalensi infeksi Leucocytozoon sp. Bull. Sains vet. XV (3).

Aryani, N. 2003. Identifikasi larva nyamuk Aedes, Culex, dan Anopheles di beberapa tempat di Denpasar. SCRIPTION Fakultas Kedokteran Hewan Universitas Udayana.

Baldassarre, G. A. and E. G. Bolen. 1994. Waterfowl Ecology and Management. John Willey and Sons, Toronto.

Bennet, G. F., A. D. Smith, W. Whitman, and M. Cameron. 1975. Hematozoa of the anatidae of the atlantic flyway II. The maritime provinces of Canada. J. Wildl. Dis 11(2) : 280 – 289 (Abs).

Clark, G. W. 1980. Hematozoa of mallard ducks (Anas platyrhynchos) of the Pacific Flyway, Washington. J. Wildl. Dis. 16(4) : 529-531 (Abs).

Coles, E. H. 1980. Veterinary clinical pathology. 3rd Ed. W.B. Saunders and Co.Philadelphia, USA.

Darmono, S. Partoutomo, Sukarsih, and R. Soetejo. 1982. Parasit darah pada ayam kampung di tiga kecamatan Kabupaten Bogor. Proceeding Seminar Penelitian Pertanian. Cisarua. Pusat Penelitian dan Pengembangan Peternakan, Badan penelitian dan Pengembangan Pertanian. Departemen Pertanian : p. 518-521.

Demsey, S and M. Kennedy. 1999. Avian Hematology. (www.spjc.edu/hec/vettech/VTDE/avianhemo/avianhemo.htm-25k) 22-9-2003

Domestic waterfowl club.2003. Light Duck Breeds… to locate via picture. The Domestic Waterfowl Club of Great Britain. (http://www.domestic-waterfowl.co.uk/page3.htm-37k) 25/12 2003

Fludge, A. M. 1997. Avian blood panel interpretation (www.californiaavianlab.com)

Gunawan, M., N. Wetta, and W. Karta. 1984. Leucocytozoonosis pada ayam kampung selama 6 tahun di Bali. Bulletin Veteriner 1 (8) : 4 (Abs.).

Houwen, B. 2000. Blood film preparation and staining procedures. Lab. Hematol. 6 : 1-7 (www.bloodline.net) 14-4-2002

Jain, N. C. 1986. Schalm’s Veterinary Hematology. 4th Ed. Lea and Febiger, USA. :

Jones, M. P. 1999. Avian clinical pathology. Vet. Clin. North Am. Exotic Anim. Pract. 2 : 663-687.

Loven, J. S., E. G. Bolen, and B. W. Cain. 1980. Blood parasitemia in South Texas wintering waterfowl population. J. Wildl. Dis. 16(1) : 25-28 (Abs.).

Matram, B. 1984. Pengaruh Imbangan Kalori/Protein dan Pembatasan Ransum Terhadap Pertumbuhan dan Produksi Telur Itik Bali. DOCTORAL DISERTATION Pajajaran University, Bandung.

Mirah Adi, Winaya, I.B.O., Utama, I. H., Suarsana, I. N., Erawan, I G. N. K., Apsari, I A., dan Hayashi, Y. 2004. Pathological abnormalities of Bali ducks sampled from traditional farming system in Bali. Jurnal Veteriner 5 (2) : 40 – 46.

Nayar, J. K., J. W. Knight, and S. R. Telford Jr. 1998. Vector ability of mosquitoes for isolates of Plasmodium elongatum from raptors in Florida. J. Parasitol. 84(3) : 542-546 (Abs.).

O’Dell, J. P. and L. W. Robbins. 1994. Hematozoa of wood ducks (Aix sponsa) in Missouri. J. Wildl. Dis. 30(1) : 36 – 39 (Abs.).

Papas, A. A. 2002. HemPath Descriptions-1 ( www.pathophysiology.uams.edu/Fall02/Hematopathology/HemPath1Dscrp.doc )

Phillips, K. M. 1999. Psittacine Blood Collection and Hematology: Basics for the Veterinary Practitioner. (www.vet.uga.edu/ivcvm/1999/Phillips/phillips.htm-48k) 12-10-2003

Qureshi,M.A. 1998. Role of Macrophages in Avian Health and Disease. Poultry Sci. 77:978-982.

Rasyaf, M. 1982. Beternak Itik. Kanisius Press, Yogyakarta.

Steel, R. G. D. and J. H. Torrie. 1991. Prinsip dan Prosedur Statistika, Suatu Pendekatan Biometrik. Translated by : B. Sumantri. Gramedia, Jakarta.

Thul, J. E., D. J. Forrester, and E. C. Greiner. 1980. Hematozoa of wood ducks (Aix spons) in the Atlantic flyway. J. Wildl. Dis. 16(3) : 383-390 (Abs.).

Williams, N. A., B. K. Calverley and J. L. Mahrt. 1977. Blood parasites of mallard and pintail ducksfrom central Alberta and the Mackenzie Delta, Northwest Territories. J. Wildl. Dis. 13(3) : 226 – 229 (Abs.).

Winarsih, W., and S. D. Widhyari. 1998. Prevalensi infeksi parasit darah pada ayam kedu (Studi kasus di Kabupaten Temanggung Jawa Tengah). Hemera Zoa 80 : 38 – 41.