International Journal of Innovation and Applied Studies ISSN 2028-9324 Vol. 8 No. 1 Sep. 2014, pp. 185-194 © 2014 Innovative Space of Scientific Research Journals http://www.ijias.issr-journals.org/ Corresponding Author: Riasat Wasee Ullah 185 Isolation and characterization of lineage-IV Peste des Petits Ruminants (PPR) virus strains from Pakistan Aamer Bin Zahur 1 , Hamid Irshad 1 , Asma Latif 1 , Aman Ullah 1 , Riasat Wasee Ullah 1 , Muhammad Afzal 2 , Sarosh Majid Salaria 3 , Zafar-ul-Ahsan Qureshi 4 , Khawar Mahboob 4 , and Shahida Afzal 4 1 Animal Health Research Laboratories, Animal Sciences Institute, National Agricultural Research Centre, Islamabad, Pakistan 2 Progressive Control of PPR in Pakistan, FAO Pakistan (GCP/PAK/127/USA) 3 Azad Jammu and Kashmir Medical College, Muzaffarabad, Azad Kashmir, Pakistan 4 Veterinary Research Institute, Lahore, Pakistan Copyright © 2014 ISSR Journals. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. ABSTRACT: A total of 62 Peste des Petits ruminants (PPR) outbreaks in sheep and goat flocks were investigated in Pakistan during 2005-2007. The presence of PPR virus (PPRV) was confirmed by clinical picture, necropsy examination, Ic-ELISA, virus isolation and RT-PCR. Of 397 tissue samples, 65% tested positive by Ic-ELISA. Six PPR virus isolates were obtained through cell culture on VERO or GKC cell from 61Ic-ELISA positive samples identified by characteristic CPEs and confirmed by testing the cell culture supernatant by Ic-ELISA and RT-PCR using PPRV specific F gene based primers. The sequence data of F gene from 6 isolates was analyzed for identities and a phylogenetic tree was generated based on 372bp F gene sequences of PPRV. The isolates were clustered into lineage 4 along with other Asian isolates. The recent isolates and a previous isolate from Pakistan (PAK-2004) were found to be monophyletic having close relationship with an Indian isolate (IND-PON). KEYWORDS: PPR, isolation, Pakistan, phylogenetic analysis, Ic-ELISA. 1 INTRODUCTION Peste des Petits Ruminants (PPR) is an “acute, highly contagious and frequently fatal viral disease of sheep, goats and wild small ruminants” [1]. The disease is characterized by fever, muco-purulent ocular and nasal discharges, necrotizing and erosive stomatitis, severe enteritis and pneumonia leading to death [2]. PPR is a trans-boundary animal disease (TAD) of significant economic importance [3], ranking among the top ten diseases affecting small ruminants [4]. It has been described as the most important single cause of morbidity and mortality in small ruminant in Nigeria [5]. The causal agent, PPR virus (PPRV), is an enveloped ribonucleic acid virus of the genus Morbillivirus of family Paramyxoviridae[6]. The disease was first reported in Cote d’Ivoire in 1942 [7]. At present, it is endemic across the Sub-Saharan Africa, Morocco, Arabian Peninsula, Middle East, Turkey, Iran, Iraq, Pakistan, India, Bangladesh, Nepal, Tajikistan and Kazakhstan in Central Asia and Tibet, China [8-12]. The disease was first recorded in Pakistan during 1991 [13]. However, there are limited reports, which have attempted to document the prevalence and demonstrated continuing activity of PPRV in the country [14-16]. Bearing in mind the impact of PPR in the background of subsistence farming, the present study was conducted with the objective to isolate and characterize local PPR virus strains circulating among small ruminants population of Pakistan with special emphasis on monitoring the changes in virulence.