1,720,986 research outputs found
Full-genome analysis of a new Italian genotype of Infectious bursal disease virus.
No full textInfectious bursal disease virus (IBDV) causes a worldwide, contagious disease of young chickens, also known as “Gumboro disease”. Since its appearance in 1965, the disease has become endemic in Italy. In a recent study an emerging IBDV genotype (ITA) was detected in IBDV-live vaccinated broilers without IBD clinical signs. VP2 sequence analysis showed that strains of the ITA genotype clustered separately from other reference strains, either classical or very virulent (vv) and from vaccines. In order to obtain a more exhaustive molecular characterization of IBDV ITA genotype and speculate its origin, full genome sequence of the field isolate IBDV/Italy/1829/2012 was determined and sequence, phylogenetic and recombination analysis performed. Results confirmed ITA to be a genetically distinctive IBDV genotype. Interestingly, multiple amino acid (aa) sequence alignment revealed that it shared several aa residues with vv IBDV strains, as well some virulence markers, especially in the VP1 protein. Nevertheless sequence analysis highlighted in ITA strain the presence of some residues typical of IBDV strains at low degree of virulence. Although homologous recombination and reassortant phenomena may naturally occur between different IBDV strains, no evidence of those events was found in the genome of the studied ITA strain
Molecular detection and characterization of Mycoplasma gallisepticum and Mycoplasma synoviae strains in backyard poultry in Italy
Get PDFMycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS) represent the most important avian Mycoplasma species in the poultry industry, causing considerable economic losses. In Italy, the presence of MG or MS has been investigated especially in commercial poultry farms. To our knowledge, no systematic investigations on MG or MS presence using highly specific diagnostic assays have been performed in backyard poultry. The aim of this study was to detect and molecularly characterize MG and MS strains in 11 backyard poultry flocks located in different regions of Italy. Tracheal swabs were collected and DNA was extracted. For MS, a PCR targeting a vlhA gene fragment was performed, and typing and subtyping was attempted. The presence of MG was investigated by a screening PCR, then MG typing by gene-targeted sequencing (GTS). All the amplicons were sequenced, then MG and MS dendrograms were constructed. All the flocks examined resulted Mycoplasma positive: 5 out of 11 (45.45%) were MG and MS positive, 3 (27.27%) were MG positive, and the remaining 3 (27.27%) were MS positive. The MS detections were assigned to types C, D, and F. All strains of type D belonged to subtype D1 and 2 unknown subtypes were identified. A MS sequence showed peculiar characteristics, which did not allow assignment to a known MS type or subtype. MG GTS analysis identified 6 MG strains belonging to 5 subclusters circulating in Italian backyards chicken flocks. The results of this study provide evidence of a risk for commercial poultry farms, especially in areas where backyard and commercial farms are close, suggesting the implementation of biosecurity measures
Virulence evaluation of Israeli Marek’s disease virus isolates from commercial poultry using their meq gene sequence
Get PDFFifty-seven Gallid alphaherpesvirus 2 (GaHV-2) isolates, collected during a 30-year period (1990-2019) from commercial poultry flocks affected by Marek's disease (MD), were molecularly characterised. The GaHV-2 meq gene was amplified and sequenced to evaluate the virus virulence, based on the number of PPPPs within the proline-rich repeats (PRRs) of its transactivation domain. The present illustration of virus virulence evaluation on a large scale of field virus isolates by molecular analysis exemplifies the practical benefit and usefulness of the molecular marker in commercial GaVH-2 isolates. The alternative assay of GaVH-2 virulence pathotyping is the classical Gold Standard ADOL method, which is difficult and impossible to employ on a large scale using the Specific Pathogen Free (SPF) chicks of the ADOL strains kept in isolators for two months. The phylogenetic analysis performed in the present study showed that the meq gene amino acid sequences of the 57 Israeli strains divide into 16 phylogenetic branches. The virulence evaluation was performed in comparison with 36 GaHV-2 prototype strains, previously characterised by the in vivo Gold Standard ADOL assay. The results obtained revealed that the GaHV-2 strains circulating in Israel have evolved into a higher virulence potential during the years, as the four-proline stretches number in the meq gene decreased over the investigated period, typically of very virulent virus prototypes. The present study supports the meq gene molecular markers for the assessment of field GaVH-2 strains virulence
Marek's disease viruses circulating in commercial poultry in Italy in the years 2015–2018 are closely related by their meq gene phylogeny
Get PDFMarek's disease (MD) is a lymphoproliferative disease important to the poultry industry worldwide; it is caused by Gallid alphaherpesvirus 2 (GaHV-2). The virulence of GaHV-2 isolates has shifted over the years from mild to virulent, very virulent and very virulent +. Nowadays the disease is controlled by vaccination, but field strains of increased virulence are emerging worldwide. Economic losses due to MD are mostly associated with its acute form, characterized by visceral lymphomas. The present study aimed to molecularly classify a group of 13 GaHV-2 strains detected in vaccinated Italian commercial chicken flocks during acute MD outbreaks, and to scrutinize the ability of predicting GaHV-2 virulence, according to the meq gene sequence. The full-length meq genes were amplified, and the obtained amino acid (aa) sequences were analysed, focusing mainly on the number of stretches of four proline molecules (PPPP) within the transactivation domain. Phylogenetic analysis was carried out with the Maximum Likelihood method using the obtained aa sequences, and the sequences of Italian strains detected in backyard flocks and of selected strains retrieved from GenBank. All the analysed strains showed 100% sequence identity in the meq gene, which encodes a Meq protein of 339 aa. The Meq protein includes four PPPP motifs in the transactivation domain and an interruption of a PPPP motif due to a proline-to-serine substitution at position 218. These features are typically encountered in highly virulent isolates. Phylogenetic analysis revealed that the analysed strains belonged to a cluster that includes high-virulence GaHV-2 strains detected in Italian backyard flocks and a hypervirulent Polish strain. Our results support the hypothesis that the virulence of field isolates can be suggested by meq aa sequence analysis
Genetic heterogeneity among chicken infectious anemia viruses detected in italian fowl
Get PDFChicken infectious anemia virus (CIAV) is a pathogen of chickens associated with immuno-suppression and with a disease named chicken infectious anemia. The present survey reports an epidemiological study on CIAV distribution in Italian broiler, broiler breeder and backyard chicken flocks. Twenty-five strains were detected by a specifically developed nested PCR protocol, and molecularly characterized by partial VP1 gene or complete genome sequencing. Viral DNA ampli-fication was successfully obtained from non-invasive samples such as feathers and environmental dust. Sequence and phylogenetic analysis showed the circulation of field or potentially vaccine-derived strains with heterogeneous sequences clustered into genogroups II, IIIa, and IIIb. Marker genome positions, reported to be correlated with CIAV virulence, were evaluated in field strains. In conclusion, this is the first survey focused on the molecular characteristics of Italian CIAVs, which have proved to be highly heterogeneous, implementing at the same time a distribution map of field viruses worldwide
Alteration of immunological parameters in infectious bronchitis vaccinated–specific pathogen-free broilers after the use of different infectious bursal disease vaccines
Get PDFThe vaccines currently available to control infectious bursal disease (IBD) include live-attenuated and inactivated vaccines, immune-complex vaccines, and vaccines consisting of viral constructs of herpesvirus of turkeys genetically engineered to express VP2 surface protein. To evaluate the impact of vaccines on the chicken immune system, 2 animal trials were performed in specific pathogen-free broiler chickens. In trial 1, birds were either vaccinated when they are one-day old with a dual recombinant herpes virus of turkey construct vaccine, expressing VP2 protein of (IBDV) and F protein of Newcastle disease virus, or an immune-complex IBDV vaccine or birds were not vaccinated. At 14, 28, and 35 D, the bursa of Fabricius was collected for bursa:body weight (B:BW) ratio calculation. In trial 2, birds were vaccinated when they were 1-day old according to the same protocol as trial 1, but at day 14, all groups also received a live infectious bronchitis (IB) vaccine. At 0, 7, 14, 21, and 28 days after IB vaccination, birds were tested by ELISA for IB serology and, soon after the last blood sampling, they were euthanized for collection of Harderian glands, trachea, and spleen and testing by flow cytometry for characterization of mononuclear cells. The immune-complex vaccine groups showed significantly lower B:BW ratio, lower IBV antibody titers, and higher mean percentage of CD8+ T cells in the spleen, trachea, and Harderian glands than those in the other experimental groups. The results of the in vivo trials coupled with a depth analysis of the repertoire of parameters involved in the immune response to IBD and IB vaccinations show one vaccine may influence the immune response of other vaccines included in the vaccination program
Molecular characterization of a Marek's disease virus strain detected in tumour-bearing turkeys
Get PDFMarek's disease (MD) is a lymphoproliferative disease caused by Gallid alphaherpesvirus 2 (GaHV-2), which primarily affects chickens. However, the virus is also able to induce tumours in turkeys, albeit less frequently than in chickens. This study reports the molecular characterization of a GaHV-2 strain detected in a flock of Italian meat-type turkeys exhibiting visceral lymphomas. Sequencing and phylogenetic analysis of the meq gene revealed that the turkey GaHV-2 has molecular features of high virulence and genetic similarity with GaHV-2 strains recently detected in Italian commercial and backyard chickens. GaHV-2 is ubiquitous among chickens despite vaccination, and chicken-to-turkey transmission is hypothesized due to the presence of broilers in neighbouring pens
Caratterizzazione molecolare di un ceppo mild del virus della malattia di Marek evidenziato in polli rurali con forma nervosa
No full textMarek’s disease is an important lymphoproliferative disease of chickens caused by Gallid herpesvirus 2, which is commonly referred to as Marek’s disease virus serotype 1 (MDV-1). This virus induces a range of pathological conditions including central and peripheral nervous system signs and, most frequently, visceral lymphomas.
There are very few data on the molecular characteristics and epidemiology of MDV-1 in Italy, both in commercial and in backyard chickens. For that reason feathers and organs were collected from Cochin chickens from a rural context affected
by a Marek’s disease classical nervous form. The MDV-1 strain detected by PCRs was sequenced and compared to a MDV-1 strain detected from broilers breeders affected by visceral lymphomas. The number of PPPP motif repetitions found in the transactivation domain of the Meq protein, indicated that the virus detected in Cochin chickens was mild MDV-1, while the one detected in commercial poultry was very virulent or very
virulent plus MDV-1. Clinical, macroscopic and microscopic fi ndings supported the molecular results
Comparative in vivo pathogenicity study of an ITA genotype isolate (G6) of infectious bursal disease virus
Get PDFRecently, a new genotype of infectious bursal disease virus (IBDV), named ITA, was detected in IBD-vaccinated Italian broilers. Genome characterization revealed ITA to be a genetically different IBDV, belonging to genogroup 6 according to a recently proposed IBDV classification. The currently available clinical data do not allow any definition of the degree of pathogenicity of the ITA-IBDV isolates. In the present study, a pathogenicity trial was conducted by the oral inoculation of specific-pathogen-free (SPF) chickens. Birds were housed in poultry isolators and inoculated at 35 days of age with an ITA-IBDV isolate (35 birds) or a strain belonging to the G1a genogroup as a comparison (35 birds). Control birds (25 birds) were contextually mock-inoculated with sterile water. Birds were observed daily for clinical signs and at 0, 7, 14, 21 and 28 days post-inoculation (dpi) were bled for IBDV antibody detection. At 2, 4, 7, 14, 21 and 28 dpi, five birds from each of the inoculated groups, and three from the control group, were euthanized and subjected to a post-mortem examination; the bursa:body weight and thymus:body weight ratios were calculated. Microscopic lesions of the bursa and thymus were scored on the basis of lymphoid necrosis and/or depletion or cortex atrophy, respectively. Both viruses induced a subclinical course of disease, as neither clinical signs nor mortality were recorded during the study, even in the presence of typical IBDV gross and microscopic lesions. Bursal damage, measured by the bursa:body weight ratio, was more noticeable and precocious after ITA-IBDV inoculation. Histopathology scores of the bursa, indicative of rapid lymphoid depletion, confirmed the aggressiveness of the ITA-IBDV strain in this organ. This study showed that, although the ITA-IBDV strain tested causes infection with a subclinical course, it induces severe damage to lymphoid tissues. Therefore, its circulation in birds might be a threat for the poultry industry and may jeopardize the success of the production cycle
- …
