Pigeon Paramyxovirus Research Articles

Characterization of two pigeon paramyxovirus type 1 isolates in China

For over three decades, there has been a continuing panzootic caused by a virulent variant avian paramyxovirus type 1 strain, the so-called pigeon paramyxovirus type 1. It is found primarily in racing pigeons, but it has also spread to wild birds and poultry. In this study, two pigeon paramyxovirus type 1 strains, SD12 and BJ13, obtained from diseased pigeons in China, were characterized. Phylogenetic analysis based on complete sequences allowed characterization of both strains as genotype VI, class II. Further phylogenetic analysis of a 374-nucleotide section of the fusion gene showed that SD12 fell into lineage VIbii-d and BJ13 into VIbii-f. The deduced amino acid sequence of the cleavage site of the fusion protein confirmed that both isolates contained the virulent motif 112K/RRQKR↓F117 at the cleavage site. Nevertheless, the values of intracerebral pathogenicity indices showed the SD12 isolate to be a velogenic strain and BJ13 isolate to be a mesogenic strain. The SD12 isolate was further investigated via clinical observation, RNA detection, histopathology and viral serology in experimentally infected 3-week-old chickens. It showed a mild pathological phenotype in chickens, with viral replication restricted to a few tissues. The molecular mechanism for the SD12 isolate to have a virulent motif but low levels of virulence for chickens requires further study.

HALO™-next generation image analysis for quantization of interstitial nephritis in pigeons after experimental infection with pigeon paramyxovirus-1

HALO™-next generation image analysis for quantization of interstitial nephritis in pigeons after experimental infection with pigeon paramyxovirus-1

Different regions of the newcastle disease virus fusion protein modulate pathogenicity.

Newcastle disease virus (NDV), also designated as Avian paramyxovirus type 1 (APMV-1), is the causative agent of a notifiable disease of poultry but it exhibits different pathogenicity dependent on the virus strain. The molecular basis for this variability is not fully understood. The efficiency of activation of the fusion protein (F) is determined by presence or absence of a polybasic amino acid sequence at an internal proteolytic cleavage site which is a major determinant of NDV virulence. However, other determinants of pathogenicity must exist since APMV-1 of high (velogenic), intermediate (mesogenic) and low (lentogenic) virulence specify a polybasic F cleavage site. We aimed at elucidation of additional virulence determinants by constructing a recombinant virus that consists of a lentogenic NDV Clone 30 backbone and the F protein gene from a mesogenic pigeon paramyxovirus-1 (PPMV-1) isolate with an intracerebral pathogenicity index (ICPI) of 1.1 specifying the polybasic sequence R-R-K-K-R*F motif at the cleavage site. The resulting virus was characterized by an ICPI of 0.6, indicating a lentogenic pathotype. In contrast, alteration of the cleavage site G-R-Q-G-R*L of the lentogenic Clone 30 to R-R-K-K-R*F resulted in a recombinant virus with an ICPI of 1.36 which was higher than that of parental PPMV-1. Substitution of different regions of the F protein of Clone 30 by those of PPMV-1, while maintaining the polybasic amino acid sequence at the F cleavage site, resulted in recombinant viruses with ICPIs ranging from 0.59 to 1.36 suggesting that virulence is modulated by regions of the F protein other than the polybasic cleavage site.

Experimental infection of different species of birds with pigeon paramyxovirus type 1 virus--evaluation of clinical outcomes, viral shedding, and distribution in tissues.

The virulence of pigeon paramyxovirus type 1 (PPMV-1) for different species of birds was investigated in two independent sets of experiments in which groups of pigeons, chickens, turkeys, quails, and geese (10 birds per group) were inoculated with 10(6) median embryo infectious doses of PPMV-1 isolate: 1) nonpassaged (nPPMV-1, intracerebral pathogenicity [ICPI] value = 1.27) and 2) after six passages in specific-pathogen-free chickens (pPPMV-1, ICPI = 1.46) via the oculonasal route. Naive birds were placed in contact with infected birds (two birds per group) to monitor virus transmission. Clinical observation was performed daily. Additionally, cloacal swabs, oropharyngeal swabs, and selected organ samples were collected on days 2, 4, 7, 10, and 14 postinfection and tested by real-time reverse transcriptase-PCR for estimation of viral shedding and distribution in tissues. Infected pigeons exhibited nervous and digestive tract symptoms, mortality, shedding, and transmission to contact birds. Chickens, turkeys, quails, and geese did not exhibit any clinical signs regardless of the PPMV-1 strain used for inoculation. However, in contrast to quails and geese, chickens and turkeys shed the virus via the oral cavity and cloaca, and transmission to contact birds was also observed. Viral RNA was identified in tissues collected from all pPPMV-1-infected birds, whereas negative results were obtained in the case of tissues taken from nPPMV-1-infected quails and geese. We conclude that the PPMV-1 used in this study was most virulent to pigeons, followed by chickens and turkeys, while quails and geese seem to have the highest level of innate resistance to this strain. However, passaging of PPMV-1 in chickens resulted in the increase of ICPI and noticeable but sometimes contrasting changes in the replication capacities of the virus.

A rapid method of accurate detection and differentiation of Newcastle disease virus pathotypes by demonstrating multiple bands in degenerate primer based nested RT-PCR

A rapid and accurate method of detection and differentiation of virulent and avirulent Newcastle disease virus (NDV) pathotypes was developed. The NDV detection was carried out for different domestic avian field isolates and pigeon paramyxo virus-1 (25 field isolates and 9 vaccine strains) by using APMV-I “fusion” (F) gene Class II specific external primer A and B (535bp), internal primer C and D (238bp) based reverses transcriptase PCR (RT-PCR). The internal degenerative reverse primer D is specific for F gene cleavage position of virulent strain of NDV. The nested RT-PCR products of avirulent strains showed two bands (535bp and 424bp) while virulent strains showed four bands (535bp, 424bp, 349bp and 238bp) on agar gel electrophoresis. This is the first report regarding development and use of degenerate primer based nested RT-PCR for accurate detection and differentiation of NDV pathotypes by demonstrating multiple PCR band patterns. Being a rapid, simple, and economical test, the developed method could serve as a valuable alternate diagnostic tool for characterizing NDV isolates and carrying out molecular epidemiological surveillance studies for this important pathogen of poultry.

Pigeon paramyxovirus type 1 variants with polybasic F protein cleavage site but strikingly different pathogenicity.

Newcastle disease viruses (NDV) isolated from pigeons (pigeon paramyxovirus type 1; PPMV-1) are mostly of mesogenic pathotype and characterized by a polybasic amino acid sequence motif at the fusion protein (F) cleavage site. This feature also applies to isolate R75/98 from Germany. Its genome consists of 15,192 nucleotides and it specifies an intracerebral pathogenicity index (ICPI) of 1.1, as is typical for mesogenic NDV. Recombinant R75/98 (rR75/98) derived by reverse genetics also possesses a polybasic F protein cleavage site but exhibits ICPI of 0.28, indicating a lentogenic virus. While ten virus passages of rR75/98 on embryonated chicken eggs did not result in any alteration of virus characteristics, virus which had been re-isolated from the brain of an intracerebrally inoculated chicken showed an increase in virulence, characterized by an ICPI of 0.93. Comparison of whole genome sequences of rR75/98 and re-isolated rR75/98 (RrR75/98) demonstrated only two amino acid differences, one in the F protein (N472K) and one in the polymerase protein (K2168R). This result indicates that only very few amino acid alterations are sufficient to modulate virus virulence in the presence of a polybasic amino acid sequence at the proteolytic F protein cleavage site.

Phylogenetic study based on the phosphoprotein gene of Iranian Newcastle disease viruses (NDV) isolates, 2010 -2012

BACKGROUND: Newcastle disease virus (NDV) is the causativeagent of the Newcastle disease (ND), a highly contagiousdisease in birds that causes significant economic losses to thepoultry industry worldwide. ND is endemic in Iran and outbreaksare reported regularly in commercial poultry flocks and differentspecies of birds. OBJECTIVES:The current study was carried outto characterize NDV based on phosphorprotein (P) gene fromrecent outbreaks in Iran, 2010-2012. METHODS: The P genefragment of NDV isolates of five chickens, 1 ostrich, and 1Pigeon paramyxovirus-1 was obtained by RT-PCR and sequenced.RESULTS: Phylogenetic analysis of sequences revealed thatchicken and ostrich NDV isolates were closely related andplaced in the genotype VII and Pigeon Paramyxovirus-1 waslocated in the genotype V. CONCLUSIONS:This is the first reportof Phosphoprotein gene sequences of NDV strains isolated inIran. This study will help us to understand the epidemiology andmolecular characteristics of Newcastle disease virus in Iran.

Lineage diversification of pigeon paramyxovirus effect re-emergence potential in chickens

Genotype VI-paramyxovirus (GVI-PMV1) is a major cause of epidemic Newcastle-like disease in Columbiformes. This genotype of avian paramyxovirus type 1 has diversified rapidly since its introduction into the US in 1982 resulting in two extant lineages, which have different population growth properties. Although some GVI-PMV1s replicate poorly in chickens, it is possible that variants with different replicative or pathogenic potential in chickens exist among the genetically-diverse GVI-PMV1s strains. To determine if variants of Columbiform GVI-PMV1 with different phylogenetic affiliations have distinct phenotypic properties in chickens, we investigated the replicative properties of 10 naturally circulating pigeon-derived isolates representing four subgroups of GVI-PMV1 in primary chicken lung epithelial cells and in chicken embryos. Our data demonstrate that GVI-PMV1 variants have different infection phenotypes in their chicken source host and that properties reflect subgroup affiliation. These subgroup replicative properties are consistent with observed dynamics of viral population growth.

Occurrence, characteristics and control of pigeon paramyxovirus type 1 in pigeons.

Newcastle disease (ND) is a highly contagious and devastating viral disease of poultry and other birds that has a worldwide distribution. ND in pigeons is called paramyxovirosis and is caused by antigenic "pigeon variant" of the virus (pigeon paramyxovirus type 1, PPMV-1). During PPMV-1 infections, central nervous system symptoms and sometimes high mortality are observed. In the case of infection with viscerotropic strains which exhibit specific affinity for the kidneys, the first observed sign is polyuria, and neural symptoms appear only in individual birds in the flock. Due to the similarity of symptoms of paramyxovirosis to the pigeon herpes virus infection (PHV), sodium chloride poisoning, overdose of ronidazole or vitamin B1 deficiency, it is necessary to perform laboratory tests to make a correct diagnosis. After virus isolation PPMV-1 can be detected initially by haemagglutination assay (HA). PPMV-1 can be confirmed by conventional serological tests such a haemagglutination inhibition test (HI) or molecular-based techniques. In the prophylaxis of paramyxovirosis in pigeons, inactivated vaccines are used, administered by subcutaneous injection in various prevention programs. However, vaccination should be only one component of a strategy of PPMV-1 control, on a par with effective biosecurity and proper, effective methods of prevention and diagnostics of paramyxovirosis.

In this issue – May 2014

A case report describes an attack of a mare and newborn foal by a swarm of bees, resulting in hundred of stings.1 After initial symptomatic treatment, both developed diffuse swelling of the head and were reported to be very agitated, which I think seems to be a surprisingly calm description of how they must have been feeling. The authors give details of the mechanisms of bee envenomation and suggest that in the rare fatal cases, IgE-mediated anaphylaxis is the most likely cause of death rather than toxicity. They give details of the clinical outcome of these and the rational basis of any treatments, especially as no direct scientific studies will be possible. A prospective study assessed three easily performed methods of skin surface sampling for bacterial culture of Staphylococcus isolates obtained from 27 dogs with superficial bacterial pyoderma (SBP).2 Confirmed lesions were sampled using a dry cotton swab, a saline-moistened cotton swab and a skin surface scraping. Staphylococcus spp. were isolated from lesions in all dogs. The isolates were susceptible to common empirical antimicrobials and all three of the sampling techniques gave similar results for culture. A surgical case report of two dogs describes the complication of distal flap necrosis associated with surgical reconstruction of hard palate defects with a buccal mucosal flap based on the angularis oris artery and vein.3 The authors show that the flaps were successfully salvaged by dividing the original flap pedicle and rotating the tissue to fill the remaining defect, after a vascular supply had been established from adjacent tissue. They note that the surgical outcome can be improved by removing any teeth that are likely to cause trauma to the flap's pedicle. Corynebacterium urealyticum is an uncommon cause of urinary tract infections in small animals and is difficult to treat because it is resistant to most antibacterial agents. A case report describes the outcome of a female speyed dog that had had surgery for bilateral ectopic ureters 14 weeks before presentation.4 The authors show the radiographic and ultrasound features of the tract and the carbonate apetite uroliths that were voided after linezolid treatment and change to urine-acidifying diet. The authors discuss the problems of treating a resistant organism and the possible choices for treatment. Feed intake in the first week is critical for cattle that are newly arrived in a feedlot, as they are often in negative energy and protein balance and they may have abnormal rumen function because of their time without feed during transport. This has an effect on the immune system and may lead to increases in diseases such as bovine respiratory disease (BRD) and cattle may need to be fed for longer to reach a marketable weight. In this report of a randomised trial of 4643 beef cattle given a urea-molasses supplement or left untreated, results showed that BRD incidence and production were not improved by the treatment.5 Morbidity from BRD was higher when cattle without permanent incisors were supplemented with urea-molasses compared with unsupplemented cattle, and the authors note that more research is needed in this area to work out the cause of this. They also found that BRD was higher in steers than heifers, as shown previously, and incidence was double in cattle bought from saleyards than off farms. An extensive case report describes the outcome of a domestic pigeon that developed a sterile recurring granuloma after vaccination with an oil-adjuvant, inactivated La Sota strain of Newcastle's disease.6 This was given off-label as protection against pigeon paramyxovirus-1 (PPMV-1). The authors describe the extensive management of the granuloma, which included surgical removal of foreign material, control of the pain and inflammation and protection against antimicrobial contamination. PPMV-1 is now considered endemic in Australia and vaccination is important, but the authors stress that the risks of adverse reaction, including sterile granuloma, must be considered. A retrospective review of case records of ultrasonography and necropsy outcomes of 62 koalas was used to investigate the accuracy of ultrasonography in assessing koala urogenital tract structural disease.7 The results showed high concordance, indicating that ultrasonography is an effective tool for evaluating structural disease of the koala urogenital tract, which is mainly chlamydiosis. The authors also stress the animal welfare benefits that can be made by wildlife carer groups through a scientific, evidence-based approach. A case report of an adult female koala was referred for assessment of buphthalmos and keratitis of the right eye, which was severe enough to obstruct examination of intraocular structures.8 Enucleation of the affected eye was performed and the histopathological diagnosis was anterior chamber collapse syndrome and secondary glaucoma. There is little information on ocular diseases in koalas. Anterior chamber collapse syndrome occurs in various species and is associated with early-life ocular disease or trauma. It is important to differentiate this acquired condition from a congenital malformation.

Serological and growth rate responses to the use of chicken Newcastle disease vaccines in pigeons.

In the face of an outbreak of pigeon paramyxovirus (PPMV), a vaccination response study was undertaken to determine if pigeons in Australia would produce a serological response similar to that considered protective in chickens. A vaccination study evaluated serological response and safety criteria in groups of 20 pigeons. One group served as unvaccinated controls; one group was vaccinated with a live V4 strain of Newcastle disease virus (NDV) and subsequently revaccinated 28 days later with an inactivated La Sota strain vaccine; the third group was vaccinated twice with the inactivated La Sota strain vaccine 28 days apart. Serum was collected from the birds for serology 28, 56, 120 and 196 days after each treatment. Safety of the vaccines was determined using observation of the birds and body weight change. Serology was performed using three variations of the haemagglutination inhibition (HI) test, including chicken red blood cells (RBC) with either V4 NDV or PPMV as the antigen and pigeon RBC with V4 NDV as the antigen. A commercial NDV ELISA test was also used. At 28 days after the second vaccination, the geometric mean titres were 6.8 and 7.3 for the live/inactivated vaccine regimen and the inactivated/inactivated regimen, respectively. The serological response of birds vaccinated with the inactivated/inactivated regimen was significantly greater than that of the controls for all of the serological tests used. Vaccination of pigeons with two doses of chicken NDV vaccine 28 days apart was safe and resulted in antibody levels considered protective for NDV in chickens.

Successful establishment and global dispersal of genotype VI avian paramyxovirus serotype 1 after cross species transmission

The evolutionary history of avian paramyxovirus serotype 1 (PMV1), which includes the agents of Newcastle disease (ND), is characterized by a series of strain emergence events since viruses in this family were first recognized in the 1920s. Despite the importance of ND to the poultry industry, little is known about PMV1 strain emergence events and the subsequent dispersal and evolution of new strains. The genotype VI-PMV1 was first identified in the 1980s and has been named pigeon paramyxovirus-1 (PPMV1) because of unusual host specificity with Columbiformes (Collins et al., 1996); it has been responsible for panzootics in both chickens and pigeons during that time. Here, we used evolutionary analyses to characterize the emergence of this contemporary PMV1 lineage. We demonstrate that GVI-PMV1 arose through cross-species transmission events from Galliformes (i.e. chicken) to Columbiformes, and quickly established in pigeon populations. Our studies revealed a close association between the time of viral emergence and panzootic events of this virus. The virus appeared first in Southeastern Europe and quickly spread across the European continent, which became the epicenter for global virus dissemination. With new viral gene sequences, we show that GVI-PMV1 viruses currently circulating in North America resulted from multiple invasion events from Europe, one associated with an exotic European Columbiformes species, and that extant lineages have diversified locally. This study extends our understanding of successful viral emergence subsequent to cross-species transmission and dispersal patterns of newly emerged avian viruses, which may improve surveillance awareness and disease control of this and other important avian pathogens.

Characteristics of Pigeon Paramyxovirus Serotype-1 Isolates (PPMV-1) from the Russian Federation from 2001 to 2009

Monitoring programs for highly dangerous avian diseases in the Russian Federation from 2001 to 2009 detected 77 samples that were PCR positive for avian paramyxovirus serotype-1 (APMV-1) from sick or dead feral and domestic pigeons. Nucleotide sequences of the fusion (F) gene, including a nucleotide sequence encoding the F protein cleavage site, were determined for these isolates. All of the studied isolates possessed virulent F0 protein cleavage sites (112KRKKRF117, 112RRQKRF117, or 112KRQKRF117). Intracerebral pathogenicity index (ICPI) values determined for seven of the isolates exceeded the value of 0.7 (the range from 0.8 to 1.41). Based on partial genome sequencing and phylogenetic analysis, the isolates were assigned to two individual sublineages within class II genotype VIb. It was determined that most of these Newcastle disease virus isolates (70/77) recovered from the pigeons belonged to a relatively poorly studied sublineage VIb/2. The complete nucleotide sequence of the genome for the Pigeon/Russia/Vladimir/687/05 isolate of sublineage VIb/2 was determined.

Detection and differentiation of pigeon paramyxovirus serotype-1 (PPMV-1) isolates by RT-PCR and restriction enzyme analysis

Detection and pathotyping of Newcastle disease virus (NDV) is extremely important because the appearance of virulent virus has significant economic consequences. During 1981 to 1985, infections of racing and show pigeons with an avian paramyxovirus serotype-1 (APMV-1) hit worldwide, and a panzootic occurred due to a variant form of classical NDV. On the basis of pathogenicity and monoclonal antibody binding studies, the virus was termed 'pigeon PMV-1' (PPMV-1). In the past, number of Newcastle disease outbreaks in poultry and other birds has been attributed to PPMV-1. PPMV-1 viruses are known to present difficulty when assessed by conventional in vivo pathogenicity tests. In this study, the technique of reverse transcription-polymerase chain reaction (RT-PCR) and restriction enzyme (RE) analysis was used to detect and differentiate PPMV-1 isolates of Indian origin. Restriction enzyme digestion analysis of RT-PCR-amplified fusion protein (F) gene, encoding for the cleavage activation sites of fusion protein, was carried out with restriction enzymes BglI, HhaI, HaeIII, HinfI, MboI, MspI, PvuII and StyI. A set of only four enzymes HhaI, MspI or HaeIII, MboI and BglI alone were sufficient to differentially detect APMV-1 and PPMV-1 viruses and their pathotypes. In conclusion, RT-PCR followed by RE analysis proved to be useful for detection and differentiation of APMV-1 and PPMV-1 isolates at genomic level.

Phylogenetic analysis and comparison of eight strains of pigeon paramyxovirus type 1 (PPMV-1) isolated in China between 2010 and 2012

Eight strains of pigeon paramyxovirus type 1 (PPMV-1) were isolated and identified in this study, from diseased pigeon flocks suspected to be infected with PPMV-1 in China between 2010 and 2012. These PPMV-1 isolates were purified using specific-pathogen-free (SPF) chicken embryo cells before full-length genomic sequencing. The complete genome of these isolates contained 15,192 nucleotides, similar to those of Newcastle disease virus (NDV) strains in genotypes V-XI, with the gene order 3'-NP-P-M-F-HN-L-5'. A six-nucleotide insertion was found to be located in the 5' non-coding region of the nucleoprotein gene in our eight PPMV-1 strains when compared with those of genotypes I, II, III, IV and V. The cleavage site of the fusion protein was (112)RRQKRF(117), a feature generally associated with virulent NDV strains. The structural proteins were in accordance with those of other PPMV-1 strains, with the exception of the W protein of pigeon/CHINA/LJL/100605. The length of the W protein was 227 amino acids, in common with PPMV-1 strains, whereas that of pigeon/CHINA/LJL/100605 was only 181 amino acids. Phylogenetic analysis, based on the genomic sequences and sequences of the fusion gene, revealed that our eight isolates should be classified as class II genotype VIb NDVs. To our knowledge, this is the first report to show that the strain pigeon/CHINA/LLN/110713 is similar to strains isolated abroad, but it was isolated in China, which implies that it may have been introduced to China from overseas. Differences between the Chinese and foreign strains were identified in three regions (nucleotide positions 1632-2229, 3023-3310 and 6103-6439). In addition, the values of ICPI and MDT demonstrated that PPMV-1 isolates were mesogenic or lentogenic, and virulence studies showed that these PPMV-1 strains were non-pathogenic in chickens, but they induced the generation of antibodies in vivo.

What's in a strain? Viral metagenomics identifies genetic variation and contaminating circoviruses in laboratory isolates of pigeon paramyxovirus type 1

We used next generation sequencing on random amplified viral nucleic acids to determine the genome sequence of 11 pigeon paramyxovirus type 1 (PPMV-1) isolates from Belgium (period 1998–2011). The PPMV-1 deep sequence data allowed identification of sequence variability in multiple PPMV-1 isolates, including one STOP codon in the Matrix gene which was present in 15% of the viral population of one isolate. Notably, mutations that were previously associated with pathogenicity in chickens were identified as minor sequence variants in one parent laboratory strain. A phylogenetic analysis of the consensus PPMV-1 genome sequences was performed. In addition to providing nearly complete paramyxovirus genome sequences, our sequence-independent approach identified the presence of pigeon circovirus (PiCV) sequences in four of these viral stocks. Real-time quantitative RT-PCR analysis specific for PMV-1 and PiCV showed that these contaminations were present in seven viral stocks consisting of allantoic fluids and was occasionally also detected in stocks passaged in embryonated chicken eggs. Phylogenetic analysis of the PiCV consensus genome sequences showed a circulation of PiCV covering the full genetic diversity of known PiCV.This study shows the value of novel sequence independent technologies for access to sequence information for the control of reference virus stocks and other biological materials, as co-infecting viruses or sequence variants from the original sample may persist in the stocks without being identified by the routine virus-specific diagnostic tools. The exact role of PiCV in pigeon disease – in particular Newcastle disease – and its potential interference with PPMV-1 diagnostics remains to be investigated.

The Evolution of Pigeon Paramyxovirus Type 1 (PPMV-1) in Great Britain: A Molecular Epidemiological Study

Newcastle disease (ND), caused by virulent strains of avian paramyxovirus type 1 (APMV-1), is considered throughout the world as one of the most important animal diseases. For over three decades now, there has been a continuing panzootic caused by a variant virulent APMV-1 strain, so-called pigeon paramyxovirus type 1 (PPMV-1), primarily in racing pigeons, which has also spread to wild birds and poultry. PPMV-1 isolations have been made in Great Britain every year since 1983. In this study, we have completed a comparative phylogenetic analysis based on a 374 nucleotide section of the fusion protein gene of 63 isolates of PPMV-1 that were isolated over a 26-year period; 43 of these were sequenced for this study. Phylogenetic analysis of these sequences revealed that all were closely related and placed in the genetic sublineage 4b (VIb), subdivision 4biif.

Expansion of an Exotic Species and Concomitant Disease Outbreaks: Pigeon Paramyxovirus in Free-Ranging Eurasian Collared Doves

Eurasian collared doves (Streptopelia decaocto) have expanded their range across the United States since their introduction several decades ago. Recent mortality events in Eurasian collared doves in Arizona and Montana, USA, during the winter of 2009-2010 were the result of pigeon paramyxovirus (PPMV), a novel disease agent. The first instance of mortality by this emerging infectious disease in this species occurred in Florida in 2001 with subsequent disease events in 2006 and 2008. Full diagnostic necropsies were performed on carcasses from the three states. PPMV was identified by RT-PCR and virus isolation and was sequenced to the VIb genotype of avian paramyxovirus-1 (APMV). Other APMVs are common in a variety of free-ranging birds, but concern is warranted because of the potential for commingling of this species with native birds, virus evolution, and threats to domestic poultry. Improved surveillance for wildlife mortality events and efforts to prevent introduction of non-native animals could reduce the threat of introducing new pathogens.

Characterization of pigeon paramyxoviruses (Newcastle disease virus) isolated in Kazakhstan in 2005

Isolates of Newcastle disease virus (NDV) from deceased wild and domestic pigeons in Kazakhstan were obtained from the Almaty region during 2005 and were genotypically analyzed by using reverse transcription polymerase chain reaction (RT-PCR) with primers specific to the viral fusion (F) protein gene. Part of the amplified F protein DNA product (nucleotide sequence 47-422) and the deduced amino acid sequences were compared phylogenetically with those from strains previously reported in other geographic regions. Phylogenetic analysis indicated that the Kazakhstanian pigeon paramyxovirus type 1 (PPMV-1) isolates belong to genotype VI or 4bii. To our knowledge, this is the first reported VI isolates that possess the sequences of ¹¹²GKRQKR¹¹⁶ * F¹¹⁷ within the F0 protein. The information is fundamental to improving the efficiency of control strategies and vaccine development for NDV.

Genetic Characterization of a Pigeon Paramyxovirus Type 1 Isolated from Columba livia in Uruguay

SUMMARY. The isolation and molecular characterization of pigeon paramyxovirus type 1 (PPMV-1) from a sick racing pigeon in Uruguay is reported for the first time. Hemagglutination inhibition (HI) tests were performed to detect antibodies against avian paramyxovirus serotype 1 (APMV-1), and a HI titer of 1/32 was obtained. Tracheal and cloacal swabs were processed by real-time reverse transcription-polymerase chain reaction (rRT-PCR) with the use of the National Veterinary Services Laboratory-U.S. Department of Agriculture validated matrix (M) gene assay and were positive for APMV-1. Viral isolation in embryonated chicken eggs confirmed the molecular detection of the isolate. A fragment corresponding to the 3' region of the fusion (F) protein gene was amplified by RT-PCR, and subsequently sequenced. The deduced amino acid sequence at the F protein cleavage site displayed the motif 112RRQKR/F117. Phylogenetic analysis of this part of the genome allowed the isolated virus to be grouped in the lineage VIb/ 4b, which suggests that it shares the same ecologic niche with other PPMV-1 that were found in the region, and it is not imported as other European or North American viruses.