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SEROLOGICAL SURVEILLANCE OF PORCINE REPRODUCTIVE AND RESPIRATORY SYNDROME IN PIGGERIES IN BALI

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Survei Penyakit Porcine Reproductive
and Respiratory Syndrome
pada Peternakan Babi di Bali
(SEROLOGICAL SURVEILLANCE OF PORCINE REPRODUCTIVE
AND RESPIRATORY SYNDROME IN PIGGERIES IN BALI)
I Nyoman Suartha
1
, I Made Suma Anthara
2
, I Wayan Wirata
3
, Tri Komala Sari
4
,
Ni Made Ritha Krisna Dewi
5
, I Gusti Ngurah Narendra
5
, I Gusti Ngurah Mahardika
4,5
1
Laboratorium Penyakit Dalam;
2
Laboratorium Farmakologi;
3
Laboratorium Bedah,
4
Laboratorium Virologi;
5
UPT Laboratorium Biomedika Fakultas Kedokteran Hewan
Universitas Udayana Denpasar
Jl PB Sudirman Denpasar, Telepon 081 238 05727, Email: gnmahardika@indosat.net.id
ABSTRAK
Penelitian ini bertujuan untuk mengetahui keberadaan dan beban virus porcine respiration and
reproductive syndrome (PRRS) pada peternakan babi di Bali. Sampel dikumpulkan dari 10 peternakan
babi intensif dan peternakan rakyat di Bali yang tersebar di delapan kabupaten. Jumlah sampel sera
yang dikumpulkan sebanyak 305. Antibodi dideteksi dengan enzyme linked immunosorbent assay (ELISA)
dan virus PRRS dideteksi menggunakan reverse transverse polymerase chain reaction (RT-PCR). Hasil
deteksi antibodi dan isolasi virus menunjukkan rataan persentase babi positif antibodi anti-PRRS
adalah 13,4% (14,3% pada peternakan rakyat dan 11,7% pada peternakan intensif). Rataan prevalensi
virus PRRS dengan RT-PCR adalah 8,9% (15,3% pada peternakan rakyat dan 5,6% pada peternakan
intensif). Tampaknya virus PRRS telah endemik pada peternakaan babi di Bali. Vaksinasi, manajemen
peternakan, biosafety dan tindak karantina sebaiknya diimplementasi pada peternakan babi di Bali
untuk mencegah kerugian peternak akibat PRRS.
Kata Kunci : porcine reproductive and respiratory syndrome, ELISA, RT-PCR, PRRS
ABSTRACT
This study aimed to determine the presence and burden of Porcine Respiration and Reproductive
Syndrome (PRRS) virus in pig farms in Bali. A total of 305 sera samples were collected from 10 intensive
pig farms and backyard piggeries located in eight districts, in Bali. The PRRS antibody and the virus was
detected using enzyme linked immunosorbent assay (ELISA) and transverse reverse polymerase chain
reaction (RT-PCR), respectively. The results showed that, generally the average percentage of positive
swine anti-PRRS antibody was 13.4%, 14.3%, and 11.7% in the backyard farms and commercial farms,
respectively. Whereas, the detection rate of PRRS virus was 8.9% (15.3% and 5.6% in the backyard farm
and commercial farms, respectively). It was concluded that PRRS virus is endemic in pigs, in Bali.
Vaccination, management, biosafety, and quarantine should be implemented to prevent the economic
loss due to PRRS.
Keywords : porcine reproductive and respiratory syndrome , pig, Bali, Elisa, RT-PCR
PENDAHULUAN
Industri peternakan babi berperan penting
dalam meningkatkan perekonomian masyara-
kat di beberapa bagian Indonesia. Peternakan
babi terkonsentrasi di Provinsi Sumatra Utara,
Jawa Tengah, Bali, Sulawesi Utara Nusa
Tenggara Timur, Maluku, dan Papua. Jumlah
babi di seluruh Indonesia diperkirakan enam
juta ekor (Http://database.deptan.go.id),
sebagian besar dari populasi tersebut berasal dari
peternakan rakyat.
Berbagai penyakit menular asal virus pada
babi telah menyebar di seluruh Indonesia. Hal
itu dapat terjadi akibat ketergantungan industri
peternakan babi di Indonesia dengan mitra
internasional dalam hal penyediaan produk
biologi asal babi, sperma babi unggul, dan
Jurnal Veteriner Maret 2013 Vol. 14 No. 1: 24-30
ISSN : 1411 - 8327
25
embrio, akan tetapi laporan tersebut sangat
terbatas.
Penyakit Porcine Respiration and
Reproductive Syndrome (PRRS) dilaporkan
telah menyebar di seluruh dunia dan
menimbulkan kerugian yang tinggi pada
peternakan babi (Neumann et al., 2005). Di
Indonesia, virus PRRS telah dilaporkan
terdeteksi di Sumatra Utara (Salusianto 2009),
tetapi virus PRRS belum pernah dilaporkan di
Bali yang merupakan salah satu sentra
peternakan babi di Indonesia. Dengan demikian,
PRRS penting untuk diketahui penyebarannya
di Bali.
Virus PRRS termasuk famili Arteriviridae,
ordo Nidovirales (Zimmermann et al., 1997;
Blaha 2000). Virus PRRS menyebabkan
kegagalan pernafasan pada babi neonatus,
aborsi pada induk bunting (Blaha 2000),
gangguan multisistem setelah masa sapih dan
penggemukan (Burch 2008). Penyakit ini dapat
ditularkan melalui air liur (Prickett et al.,
2008.), semen (Yaeger et al., 1993), dan alat
transpormasi yang tercemar virus (Dee et al,.
2007). Hal itu yang memudahkan penyebaran
penyakit PRRS ke peternakan di negara lain
terutama yang mengimpor semen untuk bibit
babi unggul.
Penyebaran dini dari pejantan dapat
dideteksi melalui pemeriksaan darah yang
diambil dengan teknik apus darah dari arteri
aurikularis (Broes et al,. 2007). Adanya gejala
demam pada pejantan tidak dapat digunakan
sebagai patokan untuk diagnosis PRRS (Reicks
et al. 2006).
Studi ini bertujuan untuk mengetahui
keberadaan dan beban virus PRRS pada
peternakan babi di Bali.
METODE PENELITIAN
Pengambilan Sampel
Sampel darah dengan antikoagulan
dikumpulkan dari 10 peternakan babi intensif
dan peternakan rakyat di Bali. Selanjutnya
serum dikumpulkan untuk deteksi antibodi dan
spesimen jaringan dan darah untuk isolasi virus.
Jumlah sampel serum dari babi yang secara
klinis sehat per kandang adalah 30. Sampel
jaringan diambil dari babi yang secara klinis
sakit. Babi dengan tanda-tanda klinis dicurigai
PRRS dinekropsi. Lesi pascamati dicatat dan
jaringan seperti paru-paru, hati, limpa, ginjal
dan kelenjar getah bening superfisial organ alat
reproduksi dikumpulkan dalam wadah tanpa
medium dan disimpan pada suhu -20°C untuk
pemeriksaan virus, sebagian sampel
dikumpulkan pada wadah dengan formalin 10%
untuk pemeriksaan histopatologi.
Deteksi Antibodi
Antibodi terhadap virus PRRS dideteksi
menggunakan ELISA kit komersial Anigen
PRRSV Ab ELISA 2.0
®
(Anigen Animal Genetics
Inc., Korea). Serum diencerkan 1:40 dengan
larutan pengencer yang telah tersedia. Serum
yag telah diencerkan ditambahkan masing-
masing ke dalam sumuran mikroplate kode
NHC dan PRRS yang sebelumnya telah dilapisi
dengan antigen rekombinan PRRS dan antigen
NHC. Protokol yang sama dilakukan untuk
kontrol positif dan negatif. Mikroplate
diinkubasikan pada suhu kamar selama 30
menit dan selanjutnya dicuci sebanyak lima
kali. Mikroplate ditambahkan anti-porcine-HRP
dan diinkubasikan selama 30 menit. Mikroplate
dicuci sebanyak lima kali. Substrat
ditambahkan untuk setiap sumur dan
diinkubasikan selama 15 menit. Setelah itu stop
solution ditambahkan 100 μL ke setiap sumur.
Nilai absorbansi dibaca dengan spektrometer
pada panjang gelombang 450 nm dan panjang
gelombag referensi 620 nm. Validitas uji dinilai
sesuai aturan pabrik. Cut-off criteria (S/P rasio)
dihitung sesuai dengan manual yag disediakan.
Rasio S/P yang lebih besar atau sama dengan
0,4 dianggap positif, sedangkan rasio S/P yang
kurang dari 0,4 diaggap negatif.
Deteksi Virus
Virus PRRS dideteksi dari sampel lapangan
dengan menggunakan reverse-transcriptase-
polymerase chain reaction (RT PCR). Primer
yang digunakan adalah NSP2-F5’-AAAGA-
CCAGATGGAGGAGGA-3’, NSP2-R 5’-GAG-
CTGAGTATTTTGGGCGTG-3’, ORF5-F 5’-
ATGTTGGGGAAGTGCTTGACC-3’, dan ORF5
–R 5’CTAGAGACGACCCCATTGTTCCGC-3’
(Feng et al., 2008). RNA genom diisolasi dari
serum atau jaringan menggunakan protokol
ekstraksi Trizol (Invitrogen) setelah sampel
jaringan ditambahkan proteinase K dalam SDS
2%. RT-PCR untuk deteksi virus PRRS
dilakukan dengan menggunakan Super-
Script
TM
III One-Step RT-PCR system dengan
Platinum® Taq DNA Polymerase (Invitrogen).
Kondisi reaksi dilakukan dalam 0,2 mM dNTP,
1,6 mM MgSO4, dengan konsentrasi masing-
masing primer 600 μM. Setelah penambahan 1-
3 μL sampel RNA dan enzim, tabung PCR
dimasukkan ke dalam thermocycler (GenAmp
PCR System 9700). Siklus RT-PCR yang lengkap
Suartha et al Jurnal Veteriner
26
adalah selama 60 menit pada suhu 45
O
C,
prapemanasan dan tahap aktivasi Tag-
polimerase pada suhu 95
O
C selama tujuh menit.
Selanjut sebayak 40 siklus yang masing-masing
45 detik pada suhu 94
O
C, selama 45 detik pada
suhu 50-55
O
C, dan pada suhu 72
O
C selama 60
detik. Tahap sintesis akhir pada suhu 72
O
C
selama lima menit. Setelah RT-PCR, sebanyak
10-20% produk ditambahkan dengan 1-2 μL
loading dye (Bromphenol-blue dan Cyline
Cyanol), kemudian dimasukkan ke dalam
sumur cetakan gel agarose 1%, bersamaan
dengan sampel, DNA ladder 100 bp (Ivitrogen)
juga ikut dielektroforesis. Gel agarose diwarnai
dengan 25μg/ml ethidium bromida. Produk
divisualisasikan dalam kotak UV dan
didokumentasikan menggunakan Photodoc-TI-
Hood.
HASIL DAN PEMBAHASAN
Sampel sera yang dianalisis dalam
penelitian ini sebanyak 305, dengan rincian
sebanyak 98 sampel sera dari peternakan rakyat
dan 197 sampel sera dari peternakan intensif
dari Bali. Sampel yang dari Bali diambil dari
delapan kabupaten (Tabel 1).
Tanda-tanda klinis yang diamati pada anak
babi sampel adalah diare, kekurusan, batuk,
dermatitis, dan pembengkakan limponodus
inguinalis superfisial. Informasi dari peternak
yang digunakan sebagai sampel menyatakan
babi induknya sering mengalami kluron.
Gambar gejala klinis dan temuan patologis
ditunjukkan pada Gambar 1. Lesi patologi yang
menonjol adalah pembengkakan limfo-nodus
inguinalis superfisial, cairan asites, nekrotik
limpa, nekrosis multifokal ginjal, dan konsolidasi
paru-paru.
Perubahan histopatologi (Gambar 2) yang
paling menonjol ditemukan di jaringan paru-
paru, berupa penebalan septa alveoli, eksudat
fibrinosa pada alveoli dan bronkus, dan
terbentuknya trombus pada pembuluh darah
paru-paru. Perubahan menojol pada glandula
dan limpa adalah nekrosis dan haemorrhagi.
Pada ginjal ditemukan nefritis interstitial,
trombus dan hemorhagi fokal.
Hasil deteksi antibodi dan isolasi virus
(Tabel 1 dan 2) menunjukkan rataan antibodi
anti-PRRS adalah 13,4% (14,3% pada
peternakan rakyat dan 11,7% pada peternakan
Tabel 1. Distribusi, jumlah sampel, asal serum, antibodi anti-PRRS dan deteksi virus.
Asal Tipe Jumlah Antibodi Prevalensi Positif Prevalensi
Sampel Peternakan Sampel positif antibodi Virus Virus
PRRS PRRS PRRS PRRS
Badung Rakyat 12 0 0,0% 9 75,0%
Intensif 22 4 18,2% 1 4,5%
Bangli Rakyat 23 3 13,0% 1 4,3%
Intensif 31 4 12,9% 0 0,0%
Buleleng Rakyat 14 9 64,3% 0 0,0%
Karangasem Rakyat 31 0 0,0% 3 9,7%
Klungkung Intensif 38 4 10,5% 6 15,8%
Jembrana Intensif 14 4 28,6% 1 7,1%
Tabanan Rakyat 7 1 14,3% 1 14,3%
Intensif 1 58 3 5,2% 2 3,4%
Intensif 2 34 4 11,8% 1 2,9%
Gianyar Rakyat 11 1 9,1% 1 9,1%
Total 305 41 13,4% 27 8,9%
Tabel 2. Prevalensi antibodi anti-PRRS dan deteksi virus PRRS pada peternakan rakyat dan
intensif di Bali.
Tipe Jumlah Positif Antibodi Prevalensi Virus Prevalensi
Peternakan sampel anti PRRS PRRS PRRS Virus
Antibodi Positif PRRS
Rakyat 98 14 14,3% 15 15,3%
Intensif 197 23 11,7% 11 5,6%
Suartha et al Jurnal Veteriner
27
Gambar 1. Gejala klinis kekurusan (A) dan kebengkakan limponodus superfisial inguinalis (B), nekrosis
multifokal pada ginjal (C) dan konsolidasi paru (D)
A
B
C
D
Gambar 2. A: Penebalan septa alveoli paru (pembesaran 100X), B: Limpoid nekrosis dan hemorrhagia
limpa (pembesaran 100X), C: nephritis and pembentuka thrombi (pembesaran 500X)
Gambar 3. Foto PRRS RT-PCR gel agarosa 1% yang ditambahakan dengan ethidium bromida dan
visualisasi dalam transluminator UV. A: DNA penanda 100-bp; BCD : positif virus PRRS;
E,F,G,H,I,J,K,L,O,P,Q: negatif virus PRRS
A
B
C
Jurnal Veteriner Maret 2013 Vol. 14 No. 1: 24-30
28
intensif). Rataan prevalensi virus PRRS adalah
8,9% (15,3% pada peternakan rakyat dan 5,6%
pada peternakan intensif). Gambar hasil deteksi
virus PRRS menggunakan RT-PCR disajikan
pada Gambar 3.
Laporan tentang penyakit PRRS sangat
sedikit di Indonesia. Tetapi, indikasi ke arah
penyakit tersebut seperti kegagalan reproduksi,
lambatnya pertumbuhan pascasapih dan adanya
gangguan respirasi yang sulit ditangani banyak
dilaporkan oleh peternak. Indikasi terhadap
infeksi PRRS pada peterna-kan babi telah
dilaporkan di Sumatra Utara (Salusianto, 2009).
Penelitian itu melaporkan bahwa kelompok
ternak babi memiliki penampilan yang jauh
lebih baik setelah divaksinasi PRRS dibadingkan
dengan kelompok ternak babi yang tidak
divaksiasi.
Sejarah penyakit yang diperoleh dari
peternak yaitu kegagalan reproduksi dan aborsi
secara berulang-ulang pada induk, pada anak
babi pra dan pascasapih terjadi gangguan
pernafasan, kekerdilan, dan kematian tinggi
pada saat neonatus. Gejala klinis reproduksi dan
respirasi adalah gejala yang paling umum
ditemukan pada penyakit PRRS (Dorr et al.,
2006), selain gejala gangguan pertumbuhan
pascasapih (Zimmermann et al. 1997). Babi
yang terserang PRRS sangat mudah kena
infeksi sekunder dari bakteri maupun infeksi
virus ( Dorr et al., 2007). Tanda-tanda klinis
yang teramati anak babi mengalami diare,
kekurusan, batuk, dermatitis, dan
pembengkakan limfonodus inguinalis
superfisial. Gejala yang sama juga dilaporkan
oleh Zimmermann et al. (1997). Gejala klinis
demam pada pejantan tidak dapat digunakan
sebagai patokan babi itu terinfeksi PRRS (Reicks
et al., 2006). Temuan patologis adalah cairan
asites, nekrotik limpa, nekrosis multifokal
ginjal, dan konsolidasi paru-paru.
Perubahan histopatologi yang paling
menonjol ditemukan di jaringan paru-paru yaitu
penebalan septa alveoli dengan derajat
keparahan bervariasi, eksudat fibriosa pada
alveoli dan bronkus, dan trombus pada buluh
darah paru-paru. Perubahan histopatologi
seperti di atas juga dilaporkan oleh Kubo dan
Ishikawa (2009). Pada kelenjar limfoid dan limpa
perubahan histopatologi yang menonjol adalah
lesi nekrosis dan perdarahan. Lesi nefritis
interstitial, pembentukan trombus, dan
perdarahan titik juga terlihat pada ginjal.
Data tersebut telah memberikan indikasi
kuat adanya infeksi virus PRRS. Penyakit PRRS
telah dilaporkan menyebar di seluruh dunia dan
menimbulkan kerugian ekonomi yang besar
pada peternakan babi akibat kegagalan
pernafasan pada neonatus dan aborsi pada induk
(Blaha 2000). Peneliti lain melaporkan pada babi
penderita PRRS terjadi postweaning syndrome
wasting multisystemic, kelemahan, dispnea,
limfadenopati, diare, dan ikterus (Allan dan Ellis,
2000). Penyebaran penyakit dilaporkan melalui
semen, alat angkut, muntahan, insekta, burung,
aerosol, dan air liur (Christoper-hennings et al.,
1995; Rossow et al., 1995; Torremorell et al.,
1997; Dee et al., 1995; dan 2007). Virus PRRS
dapat diisolasi dari serum 12 jam setelah infeksi
(Rossow et al., 1995), sedangkan pada babi
pejantan dilaporkan bervariasi dari satu sampai
14 hari (Christopher-Henning et al., 1995; Prieto
et al.,2004). Virus masih terdeteksi sampai lima
bulan pada eksudat kripta tonsil (Fangman et
al., 2007)
Keterlibatan virus PRRS telah terbukti
dalam penelitian serologi dan virologi. Pada
penelitian ini, hasilnya menunjukkan rataan
antibodi anti-PRRS adalah 13,4% dengan
prevalensi virus PRRS sebesar 8,9%. Hasil
tersebut mengindikasikan virus PRRS telah
beredar di Bali sejak lama. Hal yang sama
kemungkinan besar dapat berlaku untuk daerah
lain di Indonesia, karena sistem produksi
industri babi intensif di Bali sangat saling
terkait dengan peternakan babi di daerah lain
di Indonesia terutama penyediaan bibit unggul.
Sementara itu saat ini peternakan rakyat
hampir semuanya memelihara ras babi luar
negeri.
Upaya yang dapat dilakukan untuk
menghindari kerugian ekonomi tinggi pada
peternakan babi adalah para peternak
dianjurkan menerapkan manajemen
peternakan yang lebih baik dan melakukan
vaksinasi terhadap PRRS. Penutupan kandang
selama 6-9 bulan mampu menghilangkan
komtaminasi virus PRRS yang telah terjadi
pada kandang (Schaefer dan Morrison, 2007).
Penerapan sanitasi, penyemprotan alat angkut
dengan air panas mampu menekan kontaminasi
virus PRRS (Dee et al., 2007). Babi-babi
peternakan rakyat juga direkomendasikan
untuk divaksinasi untuk memutus siklus
penyebaran virus, disamping peran sosio-
kultural babi dalam komunitas tertentu,
terutama sebagai komponen utama dari jaring
pengaman sosial. Pada calon pejantan harus
dilakukan penyaringan awal utuk memastikan
bebas infeksi virus PRRS (Reicks et al., 2006).
Suartha et al Jurnal Veteriner
29
SIMPULAN
Virus PRRS telah endemik pada
peternakaan babi di Bali. RT-PCR dapat
digunakan untuk diagnosis rutin pada penyakit
PRRS.
SARAN
Vaksinasi, penerapan manajemen, biosafety
dan karantina sebaiknya diimplementasi pada
peternakan babi untuk mencegah kerugian
peternak akibat PRRS.
UCAPAN TERIMA KASIH
Penulis mengucapkan terimakasih kepada
PT Boehringer-Ingelheim Indonesia atas
dukungan dana penelitian. Dinas Peternakan
Bali dan Berbagai Kabupaten telah ikut
mendukung penelitian ini.
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Suartha et al Jurnal Veteriner
... Pig pathogens recognised in various locations in Indonesia include Porcine circovirus 2 (PCV2), Escherichia coli, porcine reproductive and respiratory syndrome virus (PRRSv), Streptococcus equi subspecies zooepidemicus (S. zooepidemicus), Erysipelothrix spp., Brucella spp. and classical swine fever virus (CSFv) (Manokaran et al. 2008, Besung 2012, Suartha et al. 2013, Veralyn et al. 2014, Leslie et al. 2015). In Papua province, endoparasite burdens were initially regarded to be a major contributing factor to disease and mortality (Putra et al. 2004). ...
... S. suis incidence was reported to be higher after infection with PRRS (Hoa et al. 2013). Although PRRS has been reported in Indonesia (Suartha et al. 2013), it remains undiagnosed in Papua. In the present study, prolonged delay between pig death and sampling could influence the survival of targeted bacteria thus lower recovery rate. ...
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Herd closure has been reported as a method to eliminate porcine reproductive and respiratory syndrome virus (PRRSV) from breeding herds. However, there is concern that while closing the herd to animal entries may aid in elimination of PRRSV, productivity of the herd may decrease, at least temporarily. Herd closure was conducted at 15 multiplication herds as part of a PRRSV eradication program beginning in December 2001. All herds tested positive for porcine reproductive and respiratory syndrome (PRRS) by ELISA prior to closure, and three were known to be infected. All herds were preloaded with gilts and closed for an average of 260 days. After closure, all farms tested negative for PRRSV by polymerase chain reaction and have remained negative for 4 years. The impact of closure was evaluated by comparing the number of pigs weaned during the 52 weeks prior to the day of closure to that achieved for the 52 subsequent weeks. Of the 15 herds, 13 had produced at least the same total number of weaned pigs at 52 weeks after closure. Number of services per week and change in farrowing rate accounted for 60% of the variability observed in total pigs weaned per week.
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Four years after the report of its discovery, porcine reproductive and respiratory syndrome virus (PRRSV) continues to challenge swine producers, veterinary practitioners, and animal health researchers in the United States. The prevalence of infection is high--60% to 80% of herds is a reasonable estimate--but the clinical effects of infection vary widely among farms. In many herds, infection is unapparent and productivity seemingly unaffected. Some infected herds report occasional respiratory disease outbreaks in young pigs, or periodic outbreaks of reproductive disease, and a few herds experience severe, chronic disease problems, particularly in young pigs. In these herds, secondary infections with viral or bacterial pathogens, particularly Salmonella choleraesuis, Streptococcus suis, or Haemophilus parasuis typically occur concurrently with PRRSV infections. Understanding why some herds undergo devastating episodes of clinical disease and others show no apparent effects is central to solving the problem of clinical PRRS for swine producers. Understanding the ecology and epidemiology of PRRSV is the key to preventing and controlling PRRSV in the future. The objective of this article is to review recent developments in these areas.
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The paper describes the specifics of the epidemiology of the Porcine Reproductive and Respiratory Syndrome (PRRS), that is its "behavior" as a communicable disease in porcine populations, and compares them to the general epidemiological characteristics of communicable diseases. This analysis shows that infection with the PRRS virus "behaves" epidemiologically both as an epidemic and as an endemic disease: on the one hand it can spread like an epidemic in naïve populations, and on the other it seems to linger on infinitely in an affected population with its clinical expression varying from farm to farm like an endemic disease. The paper tries to draw "epidemiological" conclusions on the general methods for controlling and/or eradicating the disease, and to identify areas of further research.