RESUMEN

Infectious Bronchitis Virus (IBV) is a major threat to the poultry industry worldwide, causing significant economic losses. While the virus's genetic structure is well understood, the specific strains circulating in Bolivia have remained uncharacterized until now. This study aimed to identify and characterize new IBV strains in Bolivia. Tissue samples from broilers exhibiting clinical signs of Infectious Bronchitis were screened to detect IBV using real-time RT-PCR (RT-qPCR). Positive samples with low cycle threshold (Ct) values were selected for sequencing the full S1 gene. Of the 12 samples analyzed, 10 were determined to be positive for IBV. However, only four samples yielded sufficient genetic material for sequencing and subsequent phylogenetic analysis. The results revealed the presence of GI-1 and GI-23 lineages, both belonging to genotype I (GI). The GI-1 lineage showed >99% sequence identity to the H120 and Massachusetts vaccine strains, suggesting a close relationship. In contrast, the GI-23 lineage clustered with other IBV strains, showing a distinct subclade that is genetically distant from Brazilian strains. No evidence of recombination was found. Furthermore, amino acid substitution analysis identified specific mutations in the S1 subunit, particularly in the hypervariable regions 1, 2, and 3. These mutations could potentially alter the virus's antigenicity, leading to reduced vaccine efficacy. The findings of this study highlight the importance of continued and broad genomic surveillance of circulating IBV strains and the need to improve vaccination strategies in Bolivia.

Asunto(s)

RESUMEN

Immunosuppressive diseases cause great losses in the poultry industry, increasing the susceptibility to infections by other pathogens and promoting a suboptimal response to vaccination. Among them, infectious bursal disease virus (IBDV) arises as one of the most important around the world. IBDV infects immature B lymphocytes, affecting the immune status of birds and facilitating infections by other pathogens such as avian infectious bronchitis virus (IBV). Although it has been reported that the interaction between these viruses increases IBV clinical signs, there are no actual studies about the interaction between regional circulating isolates that validate this statement. In this context, the objective of our work was to evaluate the effect of the interaction between local isolates of IBDV (belonging to genogroup 4) and IBV (lineage GI-16) in chickens. Thus, specific pathogen-free chickens were orally inoculated with IBDV genogroup (G) 4 or with PBS at 5 d of age. At 14-days postinoculation (dpi) the animals were intratracheally inoculated with a GI-16 IBV or with PBS. At multiple time points, groups of birds were euthanized and different parameters such as histological damage, viral load, lymphocyte populations and specific antibodies were evaluated. The success of IBDV infection was confirmed by the severity of bursal atrophy, viral detection, and presence of anti-IBDV antibodies. In IBV-infected animals, the presence of viral genome was detected in both kidney and bursa. The coinfected animals showed higher degree of lymphocyte infiltration in kidney, higher rate of animals with IBV viral genome in bursa at 28 dpi, and a clear decrease in antibody response against IBV at 28, 35, and 40 dpi. The results indicate that the infection with the local isolate of IBDV affects the immune status of the chickens, causing major severe damage, in response to IBV infection, which could consequently severely affect the local poultry industry.

Asunto(s)

RESUMEN

The avian infectious bronchitis virus (IBV) is a coronavirus that mutates frequently, leading to a contagious and acute disease that results in economic losses to the global poultry industry. Due to its genetic and serological diversity, IBV poses a challenge in preventing and controlling the pathogen. The full-length S1 sequence analysis identifies seven main genotypes (GI-GVII) comprising 35 viral lineages. In addition to the previously described lineage, a new GI lineage (GI-30) and two lineages from novel genotypes (GVIII-1 and GIX-1) have been described in Mexico. To prevent the spread of IBV outbreaks in a specific geographic location and select the suitable vaccine, it is helpful to genetically identify the circulating IBV types. Moreover, sequencing genomes can provide essential insights into virus evolution and significantly enhance our understanding of IBV variability. However, only genomes of previously described lineages (GI-1, GI-9, GI-13, and GI-17) have been reported for Mexican strains. Here, we sequenced new genomes from Mexican lineages, including the indigenous GI-30, GVIII-1, and GIX-1 lineages. Comparative genomics reveals that Mexico has relatively homogenous lineages (i.e., GI-13), some with greater variability (i.e., GI-1 and GI-9), and others extremely divergent (GI-30, GVIII-1, and GIX-1). The circulating lineages and intra-lineage variability support the unique diversity and dynamic of Mexican IBV.

Asunto(s)

RESUMEN

The antigenic and molecular characteristics of BR-I infectious bronchitis viruses (IBVs) isolated from Brazil are reported. IBVs isolated from commercial flocks with different clinical manifestations between 2003 and 2019 were submitted to antigenic and molecular characterization. The complete S1 glycoprotein gene of 11 field isolates was amplified and sequenced. The virus neutralization (VN) test showed 94.75% neutralization with a BR-I isolate and 30% or less against other worldwide reference strains. The nucleotide and amino acid sequence analyses revealed 84.3-100% and 83.5-100% identity among them, respectively. The identity values ranged from 57.1 to 82.6% for nucleotides and from 46.6-84.4% for amino acids compared with those of other genotypes. By phylogenetic tree analysis, the Brazilian isolates were branched into the BR-I genotype (lineage GI-11), which was differentiated from foreign reference strains. Selective pressure analyses of BR-I IBVs revealed evolution under purifying selection (negative pressure) for the complete S1 gene but four specific sites (87, 121, 279, and 542) under diversifying selection (positive pressure). Profiles of cleavage sites and potential N-glycosylation sites differed from those of other genotypes. The low molecular relationship among the Brazilian viruses and foreign serotypes was concordant with the VN test results. The low antigenic relatedness (ranging from 5.3-30% between Brazilian genotype BR-I and reference IBV serotypes of North America, Europe, and Asia) indicates that the BR-I genotype is a different serotype, referred to for the first time and hereafter as serotype BR-I. RESEARCH HIGHLIGHTSStrains of the BR-I genotype presented robust antigenic and molecular similarity.BR-I strains evolved under purifying selection mode (negative pressure).The BR-I genotype originated in Brazil and dispersed to other countries.BR-I genotype viruses can be referred to as the BR-I serotype.

Asunto(s)

RESUMEN

Infectious bronchitis virus (IBV) is a pathogen affecting poultry flocks worldwide. GI-23 is an IBV lineage with a rapid spread into different continents of the world, and it was reported for the first time in South American/Brazilian broiler farms last year. This study aimed to investigate the recent introduction and epidemic spread of IBV GI-23 in Brazil. Ninety-four broiler flocks infected with this lineage were evaluated from October 2021 to January 2023. IBV GI-23 was detected using real-time RT-qPCR, and the S1 gene hypervariable regions 1 and 2 (HVR1/2) were sequenced. S1 complete and HVR1/2 nucleotide sequence datasets were used to carry out phylogenetic and phylodynamic analyses. Brazilian IBV GI-23 strains clustered into two specific subclades (SA.1 and SA.2), both in tree branches with IBV GI-23 from Eastern European poultry-producing countries, suggesting two independent and recent introductions (around 2018). Viral phylodynamic analysis showed that the IBV GI-23 population increased from 2020 to 2021, remaining constant for one year and declining in 2022. S1 amino acid sequences from Brazilian IBV GI-23 presented specific and characteristic substitutions in the HVR1/2 for subclades IBV GI-23 SA.1 and SA.2. This study brings new insights into the introduction and recent epidemiology of IBV GI-23 in Brazil.

Asunto(s)

RESUMEN

IBV variants belonging to the GI-23 lineage have circulated since 1998 in the Middle East and have spread to several countries over time. In Brazil, the first report of GI-23 occurred in 2022. The study aimed to evaluate the in vivo pathogenicity of exotic variant GI-23 isolates. Biological samples were screening by real-time RT-PCR and classified in to GI-1 or G1-11 lineages. Interestingly, 47.77% were not classified in these lineages. Nine of the unclassified strains were sequenced and showed a high similarity to the GI-23 strain. All nine were isolated and three, were studied for pathogenicity. At necropsy, the main observations were the presence of mucus in the trachea and congestion in the tracheal mucosa. In addition, lesions on the tracheas showed marked ciliostasis, and the ciliary activity confirmed the high pathogenicity of isolates. This variant is highly pathogenic to the upper respiratory tract and can cause severe kidney lesions. This study confirm a circulation of GI-23 strain in the country and report, to first time, the isolation of an exotic variant of IBV in Brazil.

Asunto(s)

RESUMEN

Infectious Bronchitis Virus (IBV) is a highly contagious pathogen that causes a serious illness with global circulation. While there is extensive data available on the virus's existence and transmission in commercial chickens in Saudi Arabia, there is a lack of such information regarding guineafowls. Therefore, this study aimed to investigate possible IBV infection among guineafowls in the Al-Hassa Governorate of the Eastern Province, Saudi Arabia. Oropharyngeal and cloacal swabs were collected from several unvaccinated flocks of guinea fowls without respiratory clinical symptoms in November and December 2022, totaling 350 samples. Total RNA was extracted from the swab samples, and a conventional reverse transcription-polymerase chain reaction was employed to detect IBV. The results revealed varying amounts of IBV in oropharyngeal and cloacal swabs at different points in time, suggesting that IBV may be widely distributed among guineafowls without exhibiting any symptoms. These findings indicate that guineafowls could act as reservoirs, influencing the ecology and epidemiology of the disease. Notably, this study reports the first occurrence of IBV in the province of Al-Ahsa, highlighting that guineafowls have been naturally exposed to the virus. To support the development of effective vaccination techniques and control measures for the disease in Saudi Arabia, the recommendation for future research endeavors is conducting ongoing surveillance, viral isolation, sequencing, phylogenetic tree analysis, and serotype characterization of IBV in guineafowls.(AU)

Asunto(s)

RESUMEN

Infectious Bronchitis (IB) is a respiratory disease caused by a highly variable Gammacoronavirus, which generates a negative impact on poultry health worldwide. GI-11 and GI-16 lineages have been identified in South America based on Infectious Bronchitis virus (IBV) partial S1 sequences. However, full genome sequence information is limited. In this study we report, for the first time, the whole-genome sequence of IBV from Colombia. Seven IBV isolates obtained during 2012 and 2013 from farms with respiratory disease compatible with IB were selected and the complete genome sequence was obtained by NGS. According to S1 sequence phylogenetic analysis, six isolates belong to lineage GI-1 and one to lineage GVI-1. When whole genome was analyzed, five isolates were related to the vaccine strain Ma5 2016 and two showed mosaic genomes. Results from complete S1 sequence analysis provides further support for the hypothesis that GVI-1, considered a geographically confined lineage in Asia, could have originated in Colombia. Complete genome information reported in this research allow a deeper understanding of the phylogenetic evolution of variants and the recombination events between strains that are circulating worldwide, contributing to the knowledge of coronavirus in Latin America and the world.

Asunto(s)

RESUMEN

The gammacoronavirus avian infectious bronchitis virus (IBV) is a highly contagious respiratory pathogen of primary economic importance to the global poultry industry. Two IBV lineages (GI-11 and GI-16) have been widely circulating for decades in South America. GI-11 is endemic to South America, and the GI-16 is globally distributed. We obtained full-length IBV genomes from Argentine and Uruguayan farms using Illumina sequencing. Genomes of the GI-11 and GI-16 lineages from Argentina and Uruguay differ in part of the spike coding region. The remaining genome regions are similar to the Chinese and Italian strains of the GI-16 lineage that emerged in Asia or Europe in the 1970s. Our findings support that the indigenous GI-11 strains recombine extensively with the invasive GI-16 strains. During the recombination process, GI-11 acquired most of the sequences of the GI-16, retaining the original S1 sequence. GI-11 strains with recombinant genomes are circulating forms that underwent further local evolution. The current IBV scenario in South America includes the GI-16 lineage, recombinant GI-11 strains sharing high similarity with GI-16 outside S1, and Brazilian GI-11 strains with a divergent genomic background. There is also sporadic recombinant in the GI-11 and GI-16 lineages among vaccine and field strains. Our findings exemplified the ability of IBV to generate emergent lineage by using the S gene in different genomic backgrounds. This unique example of recombinational microevolution underscores the genomic plasticity of IBV in South America.

Asunto(s)

RESUMEN

Avian infectious bronchitis virus (IBV) is the etiological agent of a highly contagious disease in the poultry industry. The spike protein (S1 subunit) is responsible for the molecular diversity of the virus and many genetic types, and lineages are described worldwide. IBV genetic type I-strain 23 (GI-23) has spread across different continents (including Asia, Europe and Africa), causing multiple outbreaks and severe economic losses throughout the poultry industry in the last decade. The present study aimed to report the emergence and molecular characterization of GI-23 in South Brazil, being detected for the first time in South America. Eighty-two broiler flocks presenting clinical suspicion of infectious bronchitis were selected for this study. Tracheal, renal and intestinal samples were collected for IBV detection and genotyping. A total of 57 flocks were positive for IBV by generic RT-qPCR targeting 5' untranslated region and 31 also tested positive for GI-11 by a specific RT-qPCR targeting S1 gene for this lineage. The remaining 26 IBV-positive samples were genotyped by partial and one by complete S1 gene/protein sequencing. Phylogenetic analysis demonstrated that all of them clustered into a specific branch of the GI-23. S1 protein sequence analysis evidenced that all Brazilian GI-23 IBVs had the two characteristic amino acid substitutions A93T and S/H118P/L, but other changes were also observed, such as S37F (n = 21; 81%), G117S (n = 17, 65%), P122S (n = 16; 61%) and W71R (n = 9; 35%). This study brings new insights into the epidemiology of the IBV GI-23 in the world, highlighting its emergence and molecular characteristics in Brazil, South America.

Asunto(s)

RESUMEN

The avian infectious bronchitis virus (IBV) is a highly mutable coronavirus that causes an acute and highly contagious disease responsible for economic losses to the poultry industry worldwide. Preventing and controlling bronchitis disease is difficulted by the numerous IBV circulating types with limited antigenic cross-protection that hamper the prevention and control by heterologous vaccines. The coding region of the variable spike S1 receptor-attachment domain is used to classify IBV in 7 genotypes (GI-GVII) comprising 35 viral lineages (1-35). Knowledge of the circulating IBV types causing outbreaks in a specific geographic region is beneficial to select better the appropriate vaccine(s) and contribute to disease control. In the study, 17 avian infectious bronchitis virus strains were obtained from chickens showing signs of illness in Mexico from 2007 to 2021. We detected 4 lineages within genotype I, three already known (GI-3, GI-9, GI-13) and one newly described (GI-30). In addition, we identified 2 divergent monophyletic groups that are tentatively described as lineages of new genotypes (GVIII-1 and GIX-1). Our findings revealed that Mexico's high genetic IBV diversity results from the co-circulation of divergent lineages belonging to different genotypes. Mexican IBV lineages differ significantly from Massachusetts and Connecticut vaccine strains, indicating that the currently used vaccines may need to be updated.

Asunto(s)

RESUMEN

In this study we evaluated the effectiveness of adding serotype 793B vaccine to an immunization program in order to control the infectious bronchitis virus (IBV) GI-16 lineage. Therefore, two different experiments were performed. First, a virus cross-neutralization test was carried out, which indicated that neither the Massachusetts (Mass) nor 793B serotypes are antigenically related to the field isolate A13 (GI-16). We also performed a challenge trial to evaluate if the Mass/793B combination is more efficient than Mass/Connecticut (Conn) to protect chickens against the Argentinian variant A13. Thus, 40 chickens were organized in four groups. Chickens in Group A were vaccinated at 1 day of age with Mass serotype and then at 14 days old with Mass plus Conn serotypes. Chickens in Group B received Mass and 793B serotypes at 1 and 14 days old, respectively. Groups C and D remained unvaccinated. At 28 days of age, Groups A, B, and C were challenged with the A13 isolate, while Group D remained as the negative control. The statistical analysis of the ciliostasis evaluation, performed at 7 days postchallenge (dpch), indicated that the difference between Mass/793B and Mass/Conn was not significant (p > 0.05). However, the comparison against the negative control showed that only Group A was significantly different, suggesting a slightly better performance on blocking ciliostasis for the Mass/793B combination. On the other hand, no significant differences were observed in the viral load, quantified by reverse-transcription quantitative real-time PCR (RT-qPCR) in tracheal swabs and kidneys (at 3 and 7 dpch, respectively) between vaccinated groups. Furthermore, some amounts of the viral genome were found in both vaccinated groups that could indicate that neither the Mass/793B nor the Mass/Conn combinations totally inhibited the viral replication. Such viral replication in vaccinated chickens should seriously be taken into consideration because it could promote the selection of new variants in the future.

Nota de investigación­Evaluación de la eficacia de vacunas comerciales contra el virus de la bronquitis infecciosa (IBV) perteneciente al linaje GI-16 aislado durante un brote argentino. En este estudio se evaluó la efectividad de agregar la vacuna del serotipo 793B a un programa de inmunización para controlar al virus de la bronquitis infecciosa (con las siglas en inglés IBV) linaje GI-16. Por tanto, se realizaron dos experimentos diferentes. Primeramente, se llevó a cabo una prueba de neutralización cruzada de virus, que indicó que ni los serotipos Massachusetts (Mass) ni 793B están antigénicamente relacionados con el aislado de campo A13 (GI-16). También se realizó una prueba de desafío para evaluar si la combinación Massachussets/793B era más eficiente que Massachussets/Connecticut (Conn) para proteger a los pollos contra la variante argentina A13. De esta forma, 40 pollos se organizaron en cuatro grupos. Los pollos del Grupo A se vacunaron al día de edad con el serotipo Massachussets y luego a los 14 días con los serotipos Massachussets más Connecticut. Los pollos del Grupo B recibieron los serotipos Massachussets y 793B a los 1 y 14 días de edad, respectivamente. Los grupos C y D permanecieron sin vacunar. A los 28 días de edad, los Grupos A, B y C fueron desafiados con el aislado A13, mientras que el Grupo D permaneció como control negativo. El análisis estadístico de la evaluación de la ciliostasis, realizada a los 7 días después del desafío (dpch), indicó que la diferencia entre el tratamiento Massachussets/793B y Massachussets/Connecticut no fue significativa (P> 0.05). Sin embargo, la comparación con el control negativo mostró que solo el Grupo A fue significativamente diferente, lo que sugiere un desempeño ligeramente mejor en el bloqueo de la ciliostasis para la combinación Massachussets/793B. Por otro lado, no se observaron diferencias significativas (P> 0.05) en la carga viral, cuantificada mediante transcripción reversa y PCR cuantitativa en tiempo real de hisopos traqueales y riñones (a 3 y 7 días después del desafío, respectivamente) entre los grupos vacunados. Además, se encontraron algunas cantidades del genoma viral en ambos grupos vacunados que podrían indicar que ni las combinaciones Massachussets/793B ni Massachussets/Connecticut inhibieron totalmente la replicación viral. Esta replicación viral en pollos vacunados debe tenerse muy en cuenta porque podría promover la selección de nuevas variantes en el futuro.

Asunto(s)

RESUMEN

Infectious bronchitis virus (IBV) is one of the economically most important diseases affecting the South American poultry industry. The extensive genomic heterogeneity of IBV is a consequence of high mutation rates and recombination events followed by selection. Nucleotide heterogeneity is much higher in the S1 coding region of the relevant spike protein; thus, the S1 sequence is widely used for the IBV genetic classification in genotypes and lineages. Two main lineages (GI-11 and GI-16) extensively circulate in South American chicken flocks. The GI-11 lineage, found exclusively in South America, emerged in the 1950s and is currently the predominant lineage in Brazil and Uruguay. The GI-16 lineage emerged around 1979 and is now circulating in most South American regions. All South American countries include Massachusetts-type strains (GI-1 lineage) in the IBV vaccination programs. The GI-11 and GI-16 lineages display very low antigenic relatedness to Massachusetts vaccine strains. Because these vaccine strains may not confer complete protection against South American lineages, other vaccination strategies have been reported to control GI-11 and GI-16 outbreaks. Analysis of the few full-length genomes of South American strains highlights a complex recombination history of IBV in the continent. A broader geographic and temporal sampling is needed to understand the pattern of genetic variability and the evolutionary history of IBV variants in South America.

Estudio recapitulativo- Diversidad genética y antigénica del virus de la bronquitis infecciosa en América del Sur. El virus de la bronquitis infecciosa es una de las enfermedades económicamente más importantes que afecta a la industria avícola sudamericana. La extensa heterogeneidad genómica de este virus es una consecuencia de las altas tasas de mutación y de los eventos de recombinación seguidos por selección. La heterogeneidad de nucleótidos es mucho mayor en la región codificante S1 de la proteína de la espícula; por tanto, la secuencia S1 se usa ampliamente para la clasificación genética de este virus en genotipos y linajes. Dos linajes principales (GI-11 y GI-16) circulan ampliamente en las parvadas de pollos de América del Sur. El linaje GI-11, que hasta ahora se encuentra exclusivamente en América del Sur, surgió en la década de los aþos 1950s y actualmente es el linaje predominante en Brasil y Uruguay. El linaje GI-16 surgió alrededor de 1979 y ahora está circulando en la mayoría de las regiones de América del Sur. Todos los países de América del Sur incluyen cepas tipo Massachusetts (linaje GI-1) en los programas de vacunación contra la bronquitis infecciosa. Los linajes GI-11 y GI-16 muestran una relación antigénica muy baja con las cepas de la vacuna de Massachusetts. Debido a que estas cepas de la vacuna pueden no conferir una protección completa contra los linajes sudamericanos, se han reportado otras estrategias de vacunación para controlar los brotes de GI-11 y GI-16. El análisis de los pocos genomas completos de cepas sudamericanas destaca una compleja historia de recombinación del virus de la bronquitis en el continente. Se necesita un muestreo geográfico y temporal más amplio para comprender el patrón de variabilidad genética y la historia evolutiva de las variantes del virus de la bronquitis infecciosa en América del Sur.

Asunto(s)

RESUMEN

Avian coronaviruses, including infectious bronchitis virus (IBV) and turkey coronavirus (TCoV), are economically important viruses affecting poultry worldwide. IBV is responsible for causing severe losses to the commercial poultry sector globally. The objectives of this study were to identify the viruses that were causing outbreaks of severe respiratory disease in chickens in Trinidad and Tobago (T&T) and to characterize the strains. Swab samples were collected from birds showing severe respiratory signs in five farms on the island of Trinidad. Samples were tested for the presence of IBV, as well as avian influenza virus (AIV), Newcastle disease virus (NDV) and avian metapneumovirus (aMPV) by real-time reverse transcription polymerase chain reaction (qRT-PCR). All samples from the five farms tested negative for AIV, NDV and aMPV; however, samples from clinically affected birds in all five of the farms tested positive for IBV. Genetic data revealed the presence of TCoV in chickens on two of the farms. Interestingly, these two farms had never reared turkeys. Phylogenetic analysis showed that IBV S1 sequences formed two distinct clusters. Two sequences grouped with vaccine strains within the GI-1 lineage, whereas three sequences grouped together, but separately from other defined lineages, forming a likely new lineage of IBV. Pairwise comparison revealed that the three unique variant strains within the distinct lineage of IBV were significantly different in their S1 nucleotide coding regions from viruses in the closest lineage (16% difference) and locally used vaccine strains (>20% difference). Results also suggested that one of the samples was a recombinant virus, generated from a recombination event between a Trinidad virus of the GI-1 lineage and a Trinidad virus of the newly defined lineage. Many amino acid differences were also observed between the S1 coding regions of the circulating field and vaccine strains, indicating that the IBV vaccines may not be protective. Vaccine-challenge studies are however needed to prove this.

Asunto(s)

RESUMEN

Raising backyard birds is a common practice in Brazil, mainly in the countryside or suburban areas. However, the level of respiratory pathogens in these animals is unknown. We sampled two hundred chickens from 19 backyard flocks near commercial poultry farms and performed ELISA to Infectious Bronchitis Virus, avian Metapneumovirus, Mycoplasma synoviae and Mycoplasma gallisepticum. We evaluated the association between the predictive ability of ELISA and Hemagglutination-inhibition (HI)by comparing results from eight flocks positive to Mycoplasma gallisepticum on ELISA. Besides, we assessed essential biosecurity measures in the properties (multiple species birds, rodent control, hygienic conditions, and water quality for the bird`s consumption). We could access the vaccination program only on four properties; in three of them, the birds were supposedly vaccinated for IBV. Overall the properties had a poor score for the biosecurity measures, and the seroprevalence in backyard poultry flocks for IBV, a MPV, MS, and MG were respectively 87.5% (14/16), 89.5% (17/19), 100 (19/19) and MG 84.21% (16/19). We found low specificity and predictive value between ELISA and HI in MG analysis and a positive correlation between the presence of clinical symptoms and mean MG titers. Backyard chicken are pathogens reservoirs and pose a risk for the commercial poultry farms in the region, and further efforts of the governmental entities and private sector of poultry production should consider these information to avoid future economic losses.

Asunto(s)

RESUMEN

Raising backyard birds is a common practice in Brazil, mainly in the countryside or suburban areas. However, the level of respiratory pathogens in these animals is unknown. We sampled two hundred chickens from 19 backyard flocks near commercial poultry farms and performed ELISA to Infectious Bronchitis Virus, avian Metapneumovirus, Mycoplasma synoviae and Mycoplasma gallisepticum. We evaluated the association between the predictive ability of ELISA and Hemagglutination-inhibition (HI)by comparing results from eight flocks positive to Mycoplasma gallisepticum on ELISA. Besides, we assessed essential biosecurity measures in the properties (multiple species birds, rodent control, hygienic conditions, and water quality for the bird`s consumption). We could access the vaccination program only on four properties; in three of them, the birds were supposedly vaccinated for IBV. Overall the properties had a poor score for the biosecurity measures, and the seroprevalence in backyard poultry flocks for IBV, a MPV, MS, and MG were respectively 87.5% (14/16), 89.5% (17/19), 100 (19/19) and MG 84.21% (16/19). We found low specificity and predictive value between ELISA and HI in MG analysis and a positive correlation between the presence of clinical symptoms and mean MG titers. Backyard chicken are pathogens reservoirs and pose a risk for the commercial poultry farms in the region, and further efforts of the governmental entities and private sector of poultry production should consider these information to avoid future economic losses.(AU)

Asunto(s)

RESUMEN

Infectious bronchitis (IB) is one of the avian diseases with the greatest impact on poultry farming worldwide. In Brazil, strain BR-I (GI-11) is the most prevalent in poultry flocks. The present study aimed to develop a seminested RT-PCR assay specific for the diagnosis of BR-I IBV in Brazilian samples, targeting subunit 1 of the S gene. The detection limit of this assay was 10 copies of the IBV genome. In this study, 62.24% of 572 organ pools from the 5 regions of Brazil tested positive in a 3'UTR screening, and 84.83% were typed as BR-I IBV. BR-I was detected in the respiratory, digestive and urogenital tracts in pooled samples from all Brazilian geographical regions and in all the breeding systems analyzed. Specificity and sensitivity tests as well as phylogenetic analysis successfully confirmed the expected clustering of the sequences detected by this assay with the BR-I (GI-11) group. The nested PCR described in this study represents a suitable and valuable tool in the diagnosis, epidemiology, monitoring and vaccination decisions of IBV.

Asunto(s)

RESUMEN

A vacinação é a forma mais utilizada para prevenir a bronquite infecciosa causada pelo vírus da bronquite infecciosa das galinhas (IBV). Contudo, as vacinas convencionais são incapazes de diferenciar aves infectadas de vacinadas. No presente trabalho foi construído, caracterizado, e avaliado como candidato vacinal, um adenovírus recombinante expressando o gene N do IBV. O gene N foi clonado em um adenovírus humano tipo 5 defectivo e transfectado para as células HEK-293A para gerar rAd5_N. Após o vetor ser obtido como esperado e a confirmação da expressão da proteína N em HEK-293ª, foi realizada inoculação pela via oculo-nasal na dose de 10 7 TCID 50 /0,1mL para imunização de galinhas livres de patógenos específicos (SPF). A resposta imunológica do Ad5_N e a proteção contra o desafio ao IBV foram avaliadas e comparadas com uma vacina viva comercial. Não foram detectados anticorpos anti-IBV em aves vacinadas com o Ad5_N. A vacina comercial induziu anticorpos detectáveis a partir do 7º dia pós-vacinal. Em aves vacinadas com o Ad5_N não houve aumento na expressão de IFNγ. Neste estudo, o rAd5_N obtido não conferiu proteção contra desafio com IBV-M41. Os resultados indicam a necessidade de avaliar adenovírus recombinantes expressando outros genes do IBV.(AU)

Asunto(s)

RESUMEN

A vacinação é a forma mais utilizada para prevenir a bronquite infecciosa causada pelo vírus da bronquite infecciosa das galinhas (IBV). Contudo, as vacinas convencionais são incapazes de diferenciar aves infectadas de vacinadas. No presente trabalho foi construído, caracterizado, e avaliado como candidato vacinal, um adenovírus recombinante expressando o gene N do IBV. O gene N foi clonado em um adenovírus humano tipo 5 defectivo e transfectado para as células HEK-293A para gerar rAd5_N. Após o vetor ser obtido como esperado e a confirmação da expressão da proteína N em HEK-293ª, foi realizada inoculação pela via oculo-nasal na dose de 10 7 TCID 50 /0,1mL para imunização de galinhas livres de patógenos específicos (SPF). A resposta imunológica do Ad5_N e a proteção contra o desafio ao IBV foram avaliadas e comparadas com uma vacina viva comercial. Não foram detectados anticorpos anti-IBV em aves vacinadas com o Ad5_N. A vacina comercial induziu anticorpos detectáveis a partir do 7º dia pós-vacinal. Em aves vacinadas com o Ad5_N não houve aumento na expressão de IFNγ. Neste estudo, o rAd5_N obtido não conferiu proteção contra desafio com IBV-M41. Os resultados indicam a necessidade de avaliar adenovírus recombinantes expressando outros genes do IBV.(AU)

Asunto(s)

RESUMEN

In this study, we evaluated antibody and cell-mediated immune (CMI) responses in the mucosal and systemic compartments and protection against challenge with a nephropathogenic Brazilian (BR-I) strain of infectious bronchitis virus (IBV) in chickens submitted to a vaccination regime comprising a priming dose of heterologous live attenuated Massachusetts vaccine followed by a booster dose of an experimental homologous inactivated vaccine two weeks later. This immunization protocol elicited significant increases in serum and lachrymal levels of anti-IBV IgG antibodies and upregulated the expression of CMI response genes, such as those encoding CD8ß chain and Granzyme homolog A in tracheal and kidney tissues at 3, 7, and 11 days post-infection in the vaccinated chickens. Additionally, vaccinated and challenged chickens showed reduced viral loads and microscopic lesion counts in tracheal and kidney tissues, and their antibody and CMI responses were negatively correlated with viral loads in the trachea and kidney. In conclusion, the combination of live attenuated vaccine containing the Massachusetts strain with a booster dose of an inactivated vaccine, containing a BR-I IBV strain, confers effective protection against infection with nephropathogenic homologous IBV strain because of the induction of consistent memory immune responses mediated by IgG antibodies and TCD8 cells in the mucosal and systemic compartments of chickens submitted to this vaccination regime.

Asunto(s)