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Isolation, Identification and Characterization of the Strains of Contagious Ecthyma or Orf Virus from Goat and Sheep.

Jahangir Alam
Animal Health Research Division, BLRI, Savar, Dhaka-1341

Papia Monoura
Animal Health Research Division, BLRI, Savar, Dhaka-1341

M. Shahin Alam
Sheep and Goat Production Research Division, BLRI, Savar, Dhaka-1341

Abstract/ Executive Summary:

Present research work was under taken with the view to knowing the epidemiologic characteristics of contagious ecthyma (CE), isolation, identification & molecular characterization of CEW and continuous passage of previously isolated CEV in vero cell. A total of 164 households (20 from Mymensingh sadar Upazila, 80 from Savar Upazila and 64 from Chuadanga sadar Upazila) were surveyed. Moreover, 35 samples (scab materials) were collected from clinically affected and/or suspected goats. During survey no clinical cases were found in Mymensingh and Savar area. On the other hand clinical cases were found in Digri, Chuadanga. In Chuadanga, clinical cases of CE were found in about 23.44% households (15/64) and 23.89% (70/293) animals. Overall morbidity, mortality and case fatality was 23.89, 1.02 and 17.65%, respectively. Occurrences of CE were found to differ in age & sex of animal and the month or season of the year. Highest morbidity (29%) was found in goats of 7 months- 1 years aged group and lowest (2.05%) in goats of 2-3 years or more. Females were found more susceptible than male. From 35 scab materials CE virus (CEV) was detected in 85.71% (30/35) samples by PCR. For virus isolation, 10 samples were cultured in vero cell and 3 viruses were isolated. For molecular characterization VIL gene of 5 different CEV isolated previously were sequenced. Viruses were found closely related to each other. The identity among the goat viruses ranges from 97.2-98.6%. Orf virus strain OV-SA00 (Accession No. AY386264) was found most closely related to Bangladeshi isolate. A total of 203 bases from sheep virus was sequenced and consisted of adenine (A) 25.52%, thymine (T) 21.38%, guanine (G) 32.87% and cytosine (C) 20.23%. The GC content was found higher (53.10%) than AT content (46.90). Phylogenetically goat isolates clustered separately from sheep isolate. Ten passages of previously isolated CEV were given in vero cell and viral existence was confirmed by PCR in culture fluid of up to 4th passage.

Keywords: Epidemiology, Molecular characterization, Isolation, Orf virus

Location: Mymensingh sadar Upazila, Savar Upazila and Chuadanga sadar Upazila.

Start Date: 01-07-2011

End Date: 30-06-2012

Program Area: Animal Health and Management

Commodity: Goat

Objectives:

Research was under taken with a view to knowing the epidemiology, isolation, identification, molecular characterization and phylogenetic analysis of CEV. Besides, continous passage of previously isolated CEV in vero cell to adopt the virus was also an objective of this study.

Methodology:

Following mentioned interventions were done in the methodology: 1) Survey 2) Cells 3) Primer 4) Sample collection 5) Inoculum preparation 6) DNA extraction 7) Polymerase chain reaction (PCR) 8) Purification of PCR product 9) Sequencing 10) Phylogenetic analysis 11) Virus isolation and identification 12) Serial passage of previously isolated viruses

Source of Information: BLRI. Proceedings of Annual Research Review Workshop, 2011-2012. Bangladesh Livestock Research Institute (BLRI), Savar, Dhaka-1341.

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

Present research was under taken with a view to knowing the epidemiology, isolation, identification, molecular characterization and phylogenetic analysis of CEV. Besides, continous passage of previously isolated CEV in vero cell to adopt the virus was also an objective of this study. To achieve these objectives, survey was conducted in three locations viz. Mymensingh sadar Upazila, Savar Upazila and Chuadanga sadar Upazila with a structured pre-tested questionnaire developed. Moreover, samples especially scab materials were collected from clinically affected and/or suspected goats. For molecular characterization viral gene was sequenced and employed in analysis. From the findings of this study it may be concluded that: 1) The overall morbidity, mortality and case fatality in Digri, Chuadanga sadar was 23.89, 1.02 and 17.65% respectively. 2) Proliferative lesions in mouth, lips and gum was found in 100% (n=70) goats. Besides, proliferative lesions were also found in legs (19%), body (13%), cars (10%) of goats. However, no lesions were found in genital organs, udder and anus. 3) Occurrences of CE differed spatially. 4) Female goats were found more susceptible than male. 5) Highest morbidity was found in goats of 7 months-1 years aged group and lowest in goats of 2-3 years or more. About 79% goats were affected during January-March and 21% during October-December, 2011. 6) Molecular analysis of the VIL-10 gene of ecthyma virus revealed that goat originated viruses were much more homologous when compared to sheep virus. 7) The G+C content was higher in the genome of CEV than A+T content.

Knowledge Management: Report/Proceedings