PPRV Lineage Research Articles

Circulation of Peste des Petits Ruminants Virus Lineage-IV Haplotypes in Pakistan: implication for Control and Eradication

Peste des petits ruminants (PPR), a viral disease of small ruminants, was identified for the first time in Pakistan in 1991. Due to the availability of effective vaccine, FAO and OIE to set target of PPR control and eradication by 2030. The molecular signature of variation in the virus can assist in the control and eradication of a transboundary disease of PPR. In this study, the causative PPR virus was isolated in cell culture from suspected goats and sheep from Punjab, Pakistan. Out of swab samples, 74.2% produced cytopathic effect in cell culture and proceeded for amplification of nucleoprotein gene for genome based sequence comparison. Molecular typing showed that all of them belonged to the PPRV lineage IV. However, within lineage IV, haplotype variation was evident from the sequence analysis giving a clue of virus evolution over time in accordance with the route of disease spread. The isolated PPRVs being closely related to those from the neighboring countries of Dubai, Tajikistan and Iran. This subdivision indicates possible cross-border PPRV influx or efflux of PPRV strains through nomadism or trade from Northern areas (Tajikistan), across the south-western border of Taftan (Iran) and Dubai and other gulf countries. The study proposes joint effort by national and international animal health agencies to control and eradicate the transboundary disease of PPR in the region.

Development and Evaluation of a Nested PCR for Improved Diagnosis and Genetic Analysis of Peste des Petits Ruminants Virus (PPRV) for Future Use in Nascent PPR Eradication Programme.

Simple SummaryPeste des petits ruminants (PPR) is a highly contagious and economically important viral disease of small ruminants and a large number of wildlife species. The causative agent, PPR virus (PPRV) is mainly circulating across large areas in Africa, the Middle East and Asia and is, fortunately, currently subject to a global eradication program. A robust vaccination program along with a rapid and accurate laboratory diagnosis play a major role in the effective control and eradication of the disease. Furthermore, a genetic analysis of the circulating PPRV can help to identify the epidemiological linkage of the virus spread between neighboring countries. Here, we have developed an improved polymerase chain reaction (PCR) that can detect low viral loads and help in genetic identification, particularly in resource-limited settings.Peste des petits ruminants (PPR) is a highly contagious viral disease of small ruminants caused by PPR virus (PPRV). PPR is endemic in Asia, the Middle East and across large areas of Africa and is currently targeted for global eradication by 2030. The virus exists as four different lineages that are usually limited to specific geographical areas. However, recent reports of spread of PPRV, in particular of lineage IV viruses to infection-free countries and previously PPR endemic areas are noteworthy. A rapid and accurate laboratory diagnosis and reports on its epidemiological linkage for virus spread play a major role in the effective control and eradication of the disease. Currently, molecular assays, including conventional reverse transcription-polymerase chain reaction (RT-PCR) and real-time RT-PCR (RT-qPCR) are usually used for diagnosis of PPR while the sequencing of part of the nucleocapsid gene is usually carried out for the viral lineage identification. However, it is difficult to diagnose and sequence the genetic material if the animal excreted a low level of virus at the initial stage of infection or if the PPRV is degraded during the long-distance transportation of samples to the reference laboratories. This study describes the development of a novel nested RT-PCR assay for the detection of the PPRV nucleic acid by targeting the N-protein gene, compares the performance of the assay with the existing conventional RT-PCR and also provides good-quality DNA suitable for sequencing in order to identify circulating lineages. The assay was evaluated using cell culture propagated PPRVs, field samples from clinically infected animals and samples from experimentally infected animals encompassing all four lineages (I–IV) of PPRV. This assay provides a solution with an easy, accurate, rapid and cost-effective PPR diagnostic and partial genome sequencing for use in resource-limited settings.

Camelids and Cattle Are Dead-End Hosts for Peste-des-Petits-Ruminants Virus.

Peste-des-petits-ruminants virus (PPRV) causes a severe respiratory disease in small ruminants. The possible impact of different atypical host species in the spread and planed worldwide eradication of PPRV remains to be clarified. Recent transmission trials with the virulent PPRV lineage IV (LIV)-strain Kurdistan/2011 revealed that pigs and wild boar are possible sources of PPRV-infection. We therefore investigated the role of cattle, llamas, alpacas, and dromedary camels in transmission trials using the Kurdistan/2011 strain for intranasal infection and integrated a literature review for a proper evaluation of their host traits and role in PPRV-transmission. Cattle and camelids developed no clinical signs, no viremia, shed no or only low PPRV-RNA loads in swab samples and did not transmit any PPRV to the contact animals. The distribution of PPRV-RNA or antigen in lymphoid organs was similar in cattle and camelids although generally lower compared to suids and small ruminants. In the typical small ruminant hosts, the tissue tropism, pathogenesis and disease expression after PPRV-infection is associated with infection of immune and epithelial cells via SLAM and nectin-4 receptors, respectively. We therefore suggest a different pathogenesis in cattle and camelids and both as dead-end hosts for PPRV.

Experimental infection of cattle with wild type peste-des-petits-ruminants virus – Their role in its maintenance and spread

Peste des petits ruminants (PPR) is a major Transboundary animal disease (TADs) of sheep and goats in tropical regions caused by PPRV which can also infect cattle without any clinical signs but inducing seroconversion. However the epidemiological role of cattle in the maintenance and spread of the disease is not known. For the purposes of the present study, cattle were infected with a wild type candidate from each of the four lineages of PPRV and placed in separate boxes. Naive goats were then introduced into each specific box for the 30 days duration of the experiment. The results showed that no clinical signs of PPR were recorded in these infected cattle nor in the in-contact goats. The nasal and oral swabs remained negative. Serum from animals infected with three (3) of the wild type isolates of PPRV showed high percentage inhibition (PI % > 65%) in a cELISA. Only two animals out of three infected with the Nigeria 75/3 strain of lineage 2 (mild strain) had specific anti-PPR antibodies but with PI% values around the threshold of the test. Our findings suggest that cattle are dead-end hosts for PPRV and do not play an epidemiological role in the maintenance and spread of PPRV. In a PPR surveillance programme, cattle can serve as indicators of PPRV infection.

Neglected Hosts of Small Ruminant Morbillivirus.

Eradication of small ruminant morbillivirus (PPRV) is targeted for 2030. PPRV lineage IV is found in much of Asia and Africa. We used PPRV lineage IV strain Kurdistan/2011 in transmission trials to investigate the role of pigs, wild boar, and small ruminants as PPRV reservoirs. Suids were a possible source of infection.

First genetic characterization of Peste des Petits Ruminants from Niger: On the advancing front of the Asian virus lineage.

Peste des Petits Ruminants (PPR) is a serious transboundary infectious disease of small ruminants. The causal agent, PPR virus (PPRV), can be separated into four genetically distinct lineages using phylogenetic analysis. In recent decades, lineage IV of PPRV has dramatically extended its geographic distribution from Asia to the Middle East and to Africa, where it has progressively replaced other PPRV lineages. Lineages I and II are historically distributed in West Africa. Currently, lineage II appears to dominate the region, whereas the last recorded occurrence of lineage I dates back to 1994. Recent studies reported the presence of lineage IV in Nigeria, suggesting that this lineage is expanding in West Africa. In Niger, a close neighbour of Nigeria, PPRV has never been genetically characterized, despite reports of PPR incidence. In this study, pathological samples collected from sick goats were collected in 2013 during a suspected PPR outbreak in southern Niger close to the Nigerian border were compared to samples collected in a previous investigation in October 2001 in south-western Niger. These strains were characterized by sequencing and phylogenetic analysis to identify their genetic lineage. Our results show that in 2001, lineages I and II were cocirculating in south-western Niger, whereas the strain that caused the outbreak in 2013 belonged to lineage IV and is closely related to strains identified in Nigeria. These results confirm the progression of lineage IV in West Africa. The process of PPRV lineage replacement and its implications for the epidemiology and the control of the disease in this region are unclear and should be the subject of further studies in the field.

Fatal peste des petits ruminants disease in Chowsingha

This communication reports fatal Peste des petits ruminants (PPR) disease in Chowsingha (Tetracerus quadricornis), a member of the subfamily Bovinae and family Bovidae captured in a Zoological Park. The animals showed clinical signs of acute respiratory disease with frothy nasal discharge (1-2days) and mortality of twenty animals (80%) within 48hr. Necropsy of dead Chowsingha showed haemorrhagic patches in trachea and severe congestion of lungs and ocular mucosa. There was no characteristic lesion in the intestine. Swabs from trachea and nasal tract along with tissue samples of spleen and lung from dead animals were found positive for PPR virus based on reverse transcription-polymerase chain reaction using H gene and partial N gene-specific primers. Sequence analysis of complete H gene and partial N gene confirmed the aetiological agent as PPR virus lineage IV. The identity of the Chowsingha tissues used for PPRV isolation was confirmed by the 12S ribosomal RNA gene sequencing, and the amplified gene was analysed identically to the Chowsingha mitochondrial 12S ribosomal RNA gene. The present information of PPR in Chowsingha is the first report of PPRV lineage IV causing cross-species fatal disease in subfamily bovinae and family Bovidae. The acute manifestation of the disease indicates high susceptibility of this vulnerable wild bovid species to PPR lineage IV. This report extends host range and demands enhanced surveillance among subfamily bovinae to strengthen PPR eradication programme.

Concurrent infection of Bluetongue and Peste-des-petits-ruminants virus in small ruminants in Haryana State of India

Bluetongue (BT) and peste-des-petits-ruminants (PPR) are major transboundary diseases of small ruminant, which are endemic in India. Testing of bluetongue virus (BTV) and peste-des-petits-ruminants virus (PPRV) from recent outbreaks (2015-2016) in different regions of Haryana State of India revealed that 27.5% of the samples showed the presence of dual infection of BTV and PPRV. Analysis of Seg-2 of BTV (the serotype-determining protein) showed the presence of BTV-12w in several isolates. However, analysis of N gene fragment amplicons showed that viruses belong to lineage IV were most closely related to a pathogenic strain of PPRV from Delhi. This is the first report of co-circulation of PPRV lineage IV and bluetongue virus serotype 12 in the state.

Enhanced immunosurveillance for animal morbilliviruses using vesicular stomatitis virus (VSV) pseudotypes

The measurement of virus-specific neutralising antibodies represents the “gold-standard” for diagnostic serology. For animal morbilliviruses, such as peste des petits ruminants (PPRV) or rinderpest virus (RPV), live virus-based neutralisation tests require high-level biocontainment to prevent the accidental escape of the infectious agents. In this study, we describe the adaptation of a replication-defective vesicular stomatitis virus (VSVΔG) based pseudotyping system for the measurement of neutralising antibodies against animal morbilliviruses. By expressing the haemagglutinin (H) and fusion (F) proteins of PPRV on VSVΔG pseudotypes bearing a luciferase marker gene, neutralising antibody titres could be measured rapidly and with high sensitivity. Serological responses against the four distinct lineages of PPRV could be measured simultaneously and cross-neutralising responses against other morbilliviruses compared. Using this approach, we observed that titres of neutralising antibodies induced by vaccination with live attenuated PPRV were lower than those induced by wild type virus infection and the level of cross-lineage neutralisation varied between vaccinates. By comparing neutralising responses from animals infected with either PPRV or RPV, we found that responses were highest against the homologous virus, indicating that retrospective analyses of serum samples could be used to confirm the nature of the original pathogen to which an animal had been exposed. Accordingly, when screening sera from domestic livestock and wild ruminants in Tanzania, we detected evidence of cross-species infection with PPRV, canine distemper virus (CDV) and a RPV-related bovine morbillivirus, suggesting that exposure to animal morbilliviruses may be more widespread than indicated previously using existing diagnostic techniques.

Peste des Petits Ruminants Virus, Mauritania

Peste des Petits Ruminants Virus, Mauritania

Sequence and Phylogenetic Analyses of the Structural Genes of Virulent Isolates and Vaccine Strains of Peste Des Petits Ruminants Virus from India

Peste des petits ruminants (PPR) is an acute, highly contagious, notifiable and economically important transboundary viral disease of sheep and goats. In this study, sequence and phylogenetic analyses of structural protein genes, namely the nucleocapsid (N), the matrix (M), the fusion (F) and the haemagglutinin (H) coding sequences of virulent and vaccine strains of PPR virus (PPRV), were undertaken to determine the genetic variations between field isolates and vaccine strains. The open reading frame (ORF) of these genes of the isolates/strains was amplified by RT-PCR, cloned and sequenced. The ORF of N, M, F and H genes was 1578, 1008, 1641 and 1830 nucleotides (nt) in length and encodes polypeptides of 525, 335, 546 and 609 amino acids (aa), respectively, as reported earlier. Comparative sequence analyses of these four genes of isolates/strains were carried out with published sequences. It revealed an identity of 97.7-100% and 97.7-99.8% among the Asian lineage IV and 89.6-98.7% and 89.8-98.9% with other lineages of PPRV at nt and aa levels, respectively. The phylogenetic analyses of these isolates based on the aa sequences showed that all the viruses belonged to lineage IV along with other Asian isolates. This is in agreement with earlier observations that only PPRV lineage IV is in circulation in India since the disease was first reported. Further, sequence analysis of the thermostable/thermo-adapted vaccine strains showed no significant changes in the functional or structural surface protein-coding gene sequences. It is important to monitor the circulation of the PPRV in susceptible animals by H gene-based sequence comparisons in addition to the F gene- and N gene-based approaches to identify the distribution and spread of virus in the regular outbreaks that occur in endemic countries like India.