Massachusetts Serotype Research Articles

Infectious bronchitis in Brazilian chickens: current data and observations of field service personnel

The infectious bronchitis virus (IBV) was detected for the first time in Brazil by Hipólito in 1957 in chickens sold life in the municipal market of Belo Horizonte, MG, when commercial poultry production was just starting in that country. The Massachusetts (Mass) serotype was identified. However, the clinical disease was only observed in 1975, when poultry production was intensely growing. The extensive outbreak produced the classical condition in layers and breeders, affecting egg production and quality, whereas broilers presented respiratory and "nephritis-nephrosis" signs. The disease rapidly spread to all poultry-producing regions in the country, and in 1979, both the imports and the manufacturing of live vaccines against IB strains Mass, H120 and H52, were licensed. In 1980, inactivated vaccines were introduced. Molecular techniques, particularly PCR, started to bed in the identification of IBV. A retrospective analysis showed that, up to 1989, the main IBV strain circulating in Brazil was Mass. However, other studies shows the presence of a wide diversity of IBV strains in Brazil since the first strains were isolated, even before vaccination was introduced. Most researchers agree that the incidence of IBV different from Mass has increased, including of exclusively Brazilian genotypes, different from those described in other countries. Indeed, during the last few years, the number of genotypical variants has been much higher than that of the classical Mass serotype. Clinically, in addition of the classic presentations, atypical forms such as testicular atrophy and stones in the epidydimis associated to low fertility have been described. Serological techniques started to be used in vaccination monitoring and as a diagnostic tool. Serological response standards were developed, and have shown to be very useful to determine the expected profile in vaccination programs and when clinical disease is suspected. However, the immuno-enzymatic test ELISA is the most frequently used around the world due to its convenience. These situations led service people working in the field to suspect that vaccination programs using Mass strains were not providing the required protection because of the presence of variant strains. Some argue that this was expected, particularly in layers and breeders, because Mass-type vaccines have been used for a long time, whereas most agree that the emergence of variants is the primary cause of the increasing severity of the disease in the field. This is supported by the results using Ibv genotyping as diagnostic tool, independently of phenotype (pathotype x immune system x environment). Other argues that broiler carcass downgrading rates in processing plant are not consistent with the increase in IB clinical severity. Seroconversion in non-vaccinated flocks is acknowledged, but it occurs sporadically and not necessarily correlated with disease outbreaks. There is a general agreement that IBV has shown high variability in Brazil in terms of genotype, pathotype, and serotype. However, research should emphasize IBV phenotypical characteristics using birds as biological model.

Molecular Epidemiology of Avian Infectious Bronchitis in Brazil from 2007 to 2008 in Breeders, Broilers, and Layers

Multiple lineages of Brazilian strains from 2007 to 2008 of avian infectious bronchitis virus (IBV) were detected in flocks of breeders, broilers, and layers. Organs samples from 20 IBV-positive flocks with variable clinical signs were submitted to the partial amplification of S gene (nucleotides 726-1071) of IBV. Fifteen of the 20 sequenced strains segregated in a unique Brazilian cluster subdivided in three subclusters (Brazil 01, 02, and 03). Whereas three strains could be classified as Massachusetts (Mass) genotype, the remaining two strains, originating from flocks with reproductive and respiratory disorders, grouped within the 4/91-793B genotype, a genotype that has not been detected before in Brazil. The potential relevance of the findings to the poultry industry is discussed because the low level of identity of the sequenced part of the S gene from 17 of 20 detected field strains and the vaccines of the Massachusetts serotype used suggest that the level of cross-protection by the Massachusetts vaccines might be low.

Identification of Different Serotypes of Infectious Bronchitis Viruses in Allantoic Fluid Samples with Single and Multiplex RT-PCR

Background and Aims: Infectious bronchitis virus (IBV) causes an acute, highly contagious respiratory and kidney disease of chickens which results in significant economic losses in commercial broilers, layers and breeders. Rapid identification of IBV serotypes involved in respiratory complex is a problem in the differential diagnosis. Materials and Methods: In this study, a single and multiplex RT-PCR was used to detect group III, Massachusetts and 793/B serotypes of IBV that have been circulating in Iranian poultry industry since 1999. Results: Only seven out of 49 allantoic fluid samples showed positive results in single RTPCR using group specific primers. Two of these samples belonged to IBV positive group in VN test. Whereas; four samples belonged to H9N2 positive group and one sample belonged to IBV/AIV negative groups. The high sensitivity of single RT-PCR led to detect some of the missed IBV infections as compared with other virological tests. Three out of 7, IBV isolates identified with single RT-PCR, classified as 793/B serotype and four of them categorized as Massachusetts serotype in multiplex RT-PCR. Conclusion: The results indicated that multiplex RT-PCR method, can be a valuable test for rapid identification of Massachusetts and 793/B serotypes in complex respiratory complex infection of chickens

Molecular characterization of avian infectious bronchitis virus strains from outbreaks in Argentina (2001–2008)

Twenty infectious bronchitis virus isolates were recovered from broilers and layers in different outbreaks amongst commercial poultry flocks in different geographic regions of Argentina from 2001 to 2008. The viruses were isolated from the tracheas, lungs, and caecal tonsils of birds that were showing respiratory signs. Further analysis based on their nucleotide and amino acid sequences in hypervariable region (HVR) 1 and the intervening sequence including HVRs 1 and 2 (HVR1/2) of the S1 gene was done to determine the genetic relationships among them and reference strains. Five isolates were highly related to the Massachusetts or Connecticut serotypes, indicating the probability of the detection and isolation of vaccine strains. The other Argentinean isolates formed three separate clusters (A, B and C), distant from the vaccine serotypes, with no correlation between the generated clusters and a geographic pattern. These observations could explain the failure of the Massachusetts serotype vaccination programmes to control IBV in these flocks. In addition, the utilization of HVR1/2 and HVR1 sequences resulted in trees with similar topology but the phylogenetic relationships using HVR1/2 nucleotide sequences were better supported by higher bootstrap values. Therefore, the sequences of the HVR1/2 region are recommended for phylogenetic studies.

Molecular Characterization of Infectious Bronchitis Virus Strains Isolated from the Enteric Contents of Brazilian Laying Hens and Broilers

Infectious bronchitis virus (IBV) is the causative agent of avian infectious bronchitis, which is characterized by respiratory, reproductive, and renal signs. However, the role of IBV as an enteric pathogen in still controversial. In Brazil, antigenic groups of IBV divergent from the Massachusetts serotype used for vaccination schedules in that country have already been demonstrated. The present study aimed to assess the different genotypes of IBV in Brazilian commercial poultry flocks by partial sequencing of the S1 amino-terminus coding region using enteric contents as samples and examine their relationship with the vaccine serotype currently in use. Samples of enteric contents were taken as pools of five birds from each of 18 poultry farms (17 broiler and one laying farm) from five Brazilian states between 2002 and 2006. Birds were presenting watery diarrhea and poor general condition but were without respiratory, renal, or reproductive signs. Conventional antibacterial and anticoccidial therapies were unsuccessful and, furthermore, all samples proved negative for rotavirus, reovirus, and astrovirus. Eleven IBV samples were isolated in embryonated eggs and resulted in S1 sequences. Phylogenetic analysis showed that these segregated into an exclusive cluster, close to serotype D274, but distant from Massachusetts. Mean amino acid identity amongst these Brazilian strains was 94.07%; amongst these and serotypes D274, 4/91, and Massachusetts, mean amino acid identity was 77.17%, 69.94%, and 68.93%, respectively. In conclusion, the presence of genotype variant strains of IBV in Brazilian poultry flocks has been demonstrated and might be the reason for the unsuccessful control of IBV in Brazil. Furthermore, these results also strengthen the implications of IBV in enteric diseases of poultry.

S1 gene sequence analysis of a nephropathogenic strain of avian infectious bronchitis virus in Egypt

BackgroundInfectious bronchitis is highly contagious and constitutes one of the most common and difficult poultry diseases to control. IBV is endemic in probably all countries that raise chickens. It exists as dozens of serotypes/genotypes. Only a few amino acid differences in the S1 protein of vaccine and challenge strains of IBV may result in poor protection. Tropism of IBV includes the respiratory tract tissues, proventriculus and caecal tonsils of the alimentary tract, the oviduct and the kidney.ResultsInfectious bronchitis virus (IBV) strain closely related to Massachusetts (Mass) serotype was isolated from broiler chickens suffering from severe renal and respiratory distresses. The isolate was serologically identified by Dot-ELISA and further characterized by RT-PCR then genotyped using S1 gene sequence analysis. Alignment of the S1 sequence of the isolate with 16 IBV strains revealed high homology to isolates related to Mass serotype. Inoculation with the strain reproduced the disease in experimental 1-day-old chickens and resulted in 20% mortality, severe renal and moderate respiratory distresses. Marked histopathological changes in both kidney and trachea were observed in experimentally infected chickens. A protection study using the H120 live attenuated vaccine showed low protection rate in spite of high S1 sequence homology (97%). Protection based criteria were: virus re-isolation attempts from trachea, tracheal and renal histopathology as well as IBV antigens detection by immunofluorescent antibody technique in kidney sections.ConclusionPeriodical evaluation of cross-protective capabilities of IBV vaccine(s) versus recently recovered field isolates should be performed to ensure optimum control of IBV.

Detection of Massachusetts and Arkansas Serotypes of Infectious Bronchitis Virus in Broilers

The objective of this study was to compare the presence of the Arkansas (Ark) and Massachusetts (Mass) serotypes of infectious bronchitis virus (IBV) in the tracheas and cecal tonsils of commercial broilers after vaccination at 1 day of age by coarse spray. When given as a single serotype vaccine, the Mass strain was detected by reverse transcriptase-polymerase chain reaction (RT-PCR)-restriction fragment length polymorphism (RFLP) only in the tracheas, whereas the Ark strain was detected in both the tracheas and cecal tonsils. By in situ hybridization, the Mass and Ark nucleocapsid (Nc) genes were detected only at 7 days in the tracheas. When both strains were given in the mixed vaccine, the Mass strain was more consistently detected by RT-PCR-RFLP in the tracheas and cecal tonsils at early stages of infection (up to 14 days) and the Arkansas strain was more consistently detected at late stages of infection (21 and 28 days). By in situ hybridization, the IBV Nc gene was more consistently detected in the trachea at early stages of infection (7, 14, and 21 days) and in the cecal tonsils at late stages of infection (21, 28, and 35 days). In general, the Mass strain was more frequently recovered from the tracheal and cecal tonsil tissues at earlier stages of infection and the Ark strain was recovered at later stages of infection.

Genetic diversity of avian infectious bronchitis coronavirus strains isolated in China between 1995 and 2004

Summary.Twenty-six avian infectious bronchitis (IB) viruses (IBV) were isolated from outbreaks in chickens in China between 1995 and 2004. They were characterized by comparison with twenty-six Chinese reference strains and five other IBV strains. Chinese IBVs, which were mainly nephropathogenic, were placed into seven genotypes. Fourteen Chinese IBV isolates were placed in genotype I, having small evolutionary distances from each other. Genotype II included 6 strains that were isolated in the 1990s in China. Genotype III consisted of eight Chinese isolates that showed close relationship with Korean IBV isolates. Another eight IBV isolates clustered in genotype IV and showed larger evolutionary distances. The Massachusetts serotype was present in China in 1990s and was in a separate genotype. Two isolates, HN99 and CK/CH/LHN/00I, which might be a reisolation of vaccine strains, clustered into genotype VI. Four Chinese IBV isolates formed another genotype and showed larger evolutionary distances from other Chinese IBV genotypes (genotype VII). IBVs in same genotypes showed more than 90% amino acid sequence similarities, whereas most of the viruses in different genotypes showed less than 90%. The results showed that IBVs in China came from genetic changes both in IBV populations that existed before the advent of vaccination and in the viruses that were introduced through live vaccines. IBVs showing various genetic differences are cocirculating in China.

Isolation of avian infectious bronchitis coronavirus from domestic peafowl (Pavo cristatus) and teal (Anas)

Coronavirus-like viruses, designated peafowl/China/LKQ3/2003 (pf/CH/LKQ3/03) and teal/China/LDT3/2003 (tl/CH/LDT3/03), were isolated from a peafowl and a teal during virological surveillance in Guangdong province, China. Partial genomic sequence analysis showed that these isolates had the S-3-M-5-N gene order that is typical of avian coronaviruses. The spike, membrane and nucleocapsid protein genes of pf/CH/LKQ3/03 had >99 % identity to those of the avian infectious bronchitis coronavirus H120 vaccine strain (Massachusetts serotype) and other Massachusetts serotype isolates. Furthermore, when pf/CH/LKQ3/03 was inoculated experimentally into chickens (specific-pathogen-free), no disease signs were apparent. tl/CH/LDT3/03 had a spike protein gene with 95 % identity to that of a Chinese infectious bronchitis virus (IBV) isolate, although more extensive sequencing revealed the possibility that this strain may have undergone recombination. When inoculated into chickens, tl/CH/LDT3/03 resulted in the death of birds from nephritis. Taken together, this information suggests that pf/CH/LKQ3/03 might be a revertant, attenuated vaccine IBV strain, whereas tl/CH/LDT3/03 is a nephropathogenic field IBV strain, generated through recombination. The replication and non-pathogenic nature of IBV in domestic peafowl and teal under field conditions raises questions as to the role of these hosts as carriers of IBV and the potential that they may have to transmit virus to susceptible chicken populations.

Variation in the spike protein of the 793/B type of infectious bronchitis virus, in the field and during alternate passage in chickens and embryonated eggs

The degree of variation exhibited within the 793/B serotype (also known as 4/91 and CR88 serotypes) was investigated with nine French and 10 British isolates, collected between 1985 and 1994. The S1 part (1644 nucleotides) of the spike protein gene of the first known isolate of this serotype, FR/CR85131/85, had 95.9% to 97% nucleotide identity with the other isolates. Partial sequencing of isolates from Iran and Saudi Arabia, isolated in 2000, revealed approximately 95% nucleotide identity with European isolates, including the two live 793/B vaccinal strains, showing that they were not re-isolations of vaccinal virus. The data indicates that strains within the 793/B serotype have ≥96% nucleotide identity within the whole S1 gene and ≥93% nucleotide identity within the first 560 nucleotides, and ≥92% and ≥86% amino acid identities in the corresponding protein regions. This is similar to the identities exhibited within the Massachusetts serotype. Sequence analysis of a 793/B field isolate after passage in embryonated eggs, then in chickens and then again in eggs revealed selection for a serine and alanine at S1 amino acid position 95 in chicken-passaged and egg-passaged virus, respectively. There was no change in pathogenicity. This is the first demonstration at gene sequence level of host-driven selection for infectious bronchitis virus.

Standardization of an Enzyme-Linked Immunosorbent Assay for Detection of Infectious Bronchitis Virus Antibody.

An indirect enzyme–linked immunosorbent assay (ELISA) was developed for screening of antibody to avian infectious bronchitis virus (IBV). Antigen was prepared from whole-purified IBV Massachusetts serotype (BR 801 strain). Optimum dilution with minimum background for antigen concentration, rabbit anti-chicken conjugate and sera in developed ELISA was determined 0.1μg/ml, 1:3000 and 1:100, respectively. The optical densities (OD) were compared with a commercial ELISA kit. Also, the results of ELISA were compared to hemagglutination inhibition (HI) test. The correlation coefficient of the results of both ELISAs and HI was significant (P

Avian infectious bronchitis: characterization of new isolates from Italy.

The isolation of four new variants or serotypes of avian infectious bronchitis virus in Italy is reported. The antigenic characteristics of these strains were investigated by cross-neutralization tests with the new isolates, Fa 6881/97, AZ 27/98, AZ 20/97, and BS 216/01; two of the most common European serotypes, AZ 23/74 and CR 88121 (793B); and the classic Massachusetts M41 serotype in association with a panel of 17 specific antisera. On the basis of the results obtained, the new isolates show relevant serologic differences. In fact, the four isolates were not neutralized by antisera against the most common European and American serotypes; the AZ 20/97 isolate was partially neutralized by FA 6881/97 antiserum but not reciprocally. The closely related Fa 6881/97 and AZ 27/98 isolates can be considered rather diffused in our country because they have been isolated over 20 times in the last 3 yr in different parts of Italy. On the contrary, the AZ 20/97 and BS 216/01 isolates were reported only once so far. The reverse transcription-polymerase chain reaction showed that Fa 6881/97 isolate is related to 793B isolate, whereas AZ 27/98 and BS 216/01 isolates appeared not to be related to the most common European and Massachusetts serotypes.

Infectious disease survey of lesser prairie chickens in north Texas.

Lesser prairie chicken (Tympanuchus pallidicinctus) abundance, like that of most grassland birds, has declined rangewide for decades. Although habitat loss and degradation are likely ultimate causes for this decline, infectious agents, particularly microparasites, could be proximate contributors. No surveys of pathogenic bacteria or viruses have been published for this species. We surveyed 24 free-living lesser prairie chickens from Hemphill County, Texas (USA), for evidence of exposure to Salmonella typhimurium, S. pullorum, Mycoplasma gallisepticum, M. synoviae, Chlamydophila psittaci, and the avian influenza, Newcastle disease, infectious bronchitis, and reticuloendotheliosis viruses. Two of 18, and eight of 17 samples were seropositive for the Massachusetts and Arkansas serotypes of infectious bronchitis virus, respectively. Five of the eight positive individuals were juveniles, two of which were seropositive for both serotypes. All other serologic and genetic tests were negative. Because the ecological significance of these results is unknown, the pathogenesis, transmission, and/or population-level influences of infectious bronchitis and related avian coronaviruses for lesser prairie chickens deserves further study.

Molecular epizootiology of infectious bronchitis virus in Sweden indicating the involvement of a vaccine strain

To improve the detection and molecular identification of infectious bronchitis virus (avian coronavirus), two reverse transcriptase-polymerase chain reaction (PCR) assays were developed. As 'diagnostic PCR', a set of consensus nested primers was selected from highly conserved stretches of the nucleocapsid (N) gene. As 'phylogeny' PCR, a fragment of the spike protein gene (S1) was amplified and the PCR products were directly sequenced. To study the phylogenetic relationships of the viruses from various outbreaks, studies of molecular epizootiology were performed in Sweden, a Nordic region, where the occurrence of natural cases of the disease is relatively low and the occasional use of live vaccine(s) is well recorded and monitored. The disease appeared in the region in 1994, associated with production problems among layers of various ages. During outbreaks in 1995 and 1997, both layers and broilers were affected. To reduce losses, a live attenuated vaccine has been applied since 1997. By examining 12 cases between 1994 and 1998, molecular epizootiology revealed that, before 1997, the viruses had gene sequences very similar to strains of the Massachusetts serotype. However, comparative sequence analysis of the S1 gene revealed that the identity was not 100% to any of the strains of this serotype that we analysed. A virus related to the Dutch-type strain, D274, was also identified on one farm. Surprisingly, from 1997, the year that vaccination commenced with a live Massachusetts serotype vaccine, the majority of viruses detected had S1 sequences identical to the live Massachusetts vaccine strain. This genetic relation to the vaccine virus was also confirmed by N gene sequence analysis. The studies of molecular epizootiology reveal a strong probability that the vaccination had lead to the spread of the vaccine virus, causing various disease manifestations and a confusing epizootiological situation in the poultry population.

Resultados preliminares da utilização de cultivos de anéis de traquéia para o estudo de estirpes brasileiras do vírus da bronquite infecciosa das galinhas

Tracheal organ cultures (TOC) were prepared and used for evaluating four Brazilian isolates of infectious bronchitis virus (IBV). IBV field isolates and vaccine strains were titrated in TOC and results compared to those from chicken embrionated eggs. Serum neutralization (SN) employing IBV strain-specific serum was performed for evaluating relationships between isolates. Titration results of tests performed in TOC or eggs were in mutual agreement and were considered for validating the adapted TOC methodology as alternative for virological studies in our laboratory. Sera specific to M41 (Massachusetts) or A5968 (Connecticut) did neutralize their respective IBV strains only. Field strains 208 and 29-78 specific sera did neutralize Massachusetts serotype strains M41 and H120, but PM2 serum did only M41. Strain PM4 specific serum did not neutralize any of the reference IBV analyzed, including M41, A5968 and H120 and may indicate that the isolate is serologically different from the Massachusetts serotype, currently adopted for vaccine strains in Brazil.

Genetic and Antigenic Diversity in Avian Infectious Bronchitis Virus Isolates of the 1940s

In order to verify a commonly held assumption that only Massachusetts (Mass) serotype of infectious bronchitis virus (IBV) was prevalent in the United States between the 1930s (when IBV was first isolated) and the 1950s (when the use of commercial IBV vaccines began), we examined 40 IBV field isolates from the 1940s. Thirty-eight of those isolates were recognized as Mass serotype viruses based on their reactivity to Mass-specific monoclonal antibody (Mab) and neutralization by Mass-specific chicken serum. The remaining two isolates, N-M24 and N-M39, that did not react with Mass-specific Mab, resisted neutralization by Mass-specific chicken serum, and were neutralized only by homologous chicken antibody were identified as non-Mass IBV. When the first 900 nucleotides (nt) from the 5'-end of the spike (S1) glycoprotein gene and their deduced amino acid (aa) sequences were compared, the two non-Mass isolates differed from each other by 24% and 28%, respectively. In a similar comparison, the non-Mass viruses N-M24 and N-M39 differed from M28, a Mass-type isolate from the 1940s, by 21% and 22% (nt) and 28% and 27% (aa), respectively. These data indicate that antigenic and genetic diversity among IBV isolates existed even in the 1940s. Interestingly, when the N-terminal region of the S1 of M28 was compared to that of M41, a prototype Mass virus that has undergone countless number of in vivo and in vitro host passages, the two viruses differed by only 2% (nt) and 4% (aa). This finding suggests that frequent genetic changes are not inherent in all IBV genomes.

A nomenclature for avian coronavirus isolates and the question of species status

Currently, there is no agreed naming system for isolates of infectious bronchitis virus (IBV), whose host is the domestic fowl (Gallus gallus domesticus). A uniform, informative system for naming IBV isolates would be very helpful. Furthermore, the desirability of a single naming system has become more important with the recent discoveries that coronaviruses with genome organizations and gene sequences very similar to those of IBV have been isolated from turkeys (Meleagris gallopavo) and pheasants (Phasianus colchicus). To date, no genetic features have been found that are unique to turkey isolates and to pheasant isolates that would permit unequivocal differentiation from IBVs. Should the avian coronaviruses from turkeys, pheasants and other birds each be considered as distinct coronavirus species? Or should avian coronaviruses that have gene sequences similar to those of IBV be treated as host-range variants of IBV or, more objectively, as host-range variants of a species that might be called avian coronavirus (ACoV)? Clearly, the topic of avian coronavirus species differentiation requires debate. For the moment, a naming system for avian coronavirus isolates is overdue. Increasingly, papers will include data of coronaviruses isolated from more than one species of bird. It is desirable to have a nomenclature for avian coronaviruses that indicates the host species of origin. Furthermore, it would be helpful if the name of an isolate included the country/region of origin, an isolate number and the year of isolation. The names of avian paramyxovirus (APMV) and avian influenza virus (AIV) isolates have long since contained this information; I suggest that we adopt a similar convention for isolates of avian coronaviruses. For example, the D274 isolate of IBV could be named chicken/Netherlands/D274/78. Representatives of avian coronaviruses from turkey and pheasant would include turkey/United States(Nc)/NC95/95 and pheasant/UK/750/83. Two upper case letters would be used to denote country of isolation, whereas one upper and one lower case letter would be used to indicate state or province, e.g. Nc, North Carolina. The full-length names could be abbreviated, when desired, similar to the convention used for AIV isolates, e.g. chNL78, tyUS(Nc)95 and phUK83. If the serotype of an isolate has been clearly established, this might be included in the name at end, like the serotype designation of AIVs, e.g. chicken/China/NRZ/91 (Mass.) for the Chinese isolate of the Massachusetts serotype. This suggested naming system for isolates is essentially neutral with regard to whether viruses from different bird species should be considered as different coronavirus species or simply as variants of just one avian coronavirus species. In my opinion an informative nomenclature for avian coronavirus isolates is required now, to improve communication, and need not be delayed until a decision on the definition of coronavirus species has been made.

Molecular Characterization of an Infectious Bronchitis Virus Strain Isolated from an Outbreak in Vaccinated Layers

An infectious bronchitis virus (IBV) strain CT/7852/97 was isolated from a commercial layer flock experiencing decreased egg production and poor egg quality. Reciprocal virus neutralization test demonstrated that the isolate was closely related to Massachusetts, Arkansas, and JMK. With the use of both reverse transcription-polymerase chain reaction-restriction fragment length polymorphism and multiplex polymerase chain reaction methods, results consistently showed that CT/7852/97 isolate was Massachusetts serotype specific only. The nucleotide sequence of S1 polypeptide coding region of CT/7852/97 was determined and compared with the published sequences of 12 IBV strains from North America. The highest nucleotide homology with H120 (97%) and the least homology (62%) with Delaware (De/072/92) were observed. Massachusetts 41 and Beaudette shared 94% and 93%, respectively, nucleotide similarity with CT/7852/97, and Connecticut, JMK, and Arkansas 99 had 94%, 83%, and 78%, respectively, nucleotide similarity with CT/7852/97. Furthermore, no possible recombination site on S1 gene among Massachusetts 41, Arkansas 99, and JMK was detected in the isolate.

Morphologic Observations on Respiratory Tracts of Chickens after Hatchery Infectious Bronchitis Vaccination and Formaldehyde Fumigation

The histologic changes in the respiratory tracts of chickens were evaluated after hatchery fumigation with 40% formaldehyde vapors and vaccination against infectious bronchitis virus with live attenuated vaccine (Massachusetts serotype). One-day-old chickens were housed in four isolation units in controlled environmental conditions, fed and watered ad libitum, and separated into four groups: 1) fumigated and vaccinated birds (FV group); 2) nonfumigated and vaccinated birds (NFV group); 3) fumigated and nonvaccinated birds (FNV group); and 4) control group (C group). All birds were tested to be free from Mycoplasma gallisepticum and Mycoplasma synoviae. After necropsy on the first, eighth, and twenty-sixth days after birth, samples from tracheal upper portion and lungs were conventionally processed for light, scanning, and transmission electron microscopy. Tissue response was monitored by microscopic examination of trachea and lung. On the first day of observation, fumigated and vaccinated birds (FV group) showed extensively damaged tracheal epithelium with exfoliated areas and some active glands with electrodense granules, and in the lung, the primary bronchi epithelium had disorganized cilia and abundant lymphocytes, with emphysematous areas in tertiary bronchus. On day 8 after vaccination, cubical and cylindrical tracheal cell proliferation was observed, and on day 26, ciliated columnar epithelium was almost regenerated with heterophil corion infiltration, and hyaline cartilage nodules appeared in parabronchi. The nonfumigated and vaccinated birds (NFV) revealed less injury on the epithelial surface and a more rapid response to epithelial regeneration than the in only fumigated animals (FNV). The control group did not show remarkable morphologic changes. Postvaccinal and fumigation effects on the upper respiratory tract were temporary, whereas in lungs, increased emphysema, cartilage nodules in the interchange zone, and general lymphocyte infiltration had caused intensive injury.

Characterization of Infectious Bronchitis Viruses Isolated from Outbreaks of Disease in Commercial Flocks in Brazil

Fifteen isolations of infectious bronchitis (IB) virus were made from a total of 126 Brazilian poultry flocks of all ages that were examined. These flocks (14 chicken and 1 quail) were experiencing a variety of IB-like conditions including respiratory disease, digestive and kidney problems, and drops in egg production. One of the isolates was of the Massachusetts serotype. The remainder were examined by means of cross-neutralization tests in tracheal organ cultures and were shown to belong to at least four antigenic groups, all different from ones described previously in other countries. Some, but not all, of the flocks from which they were isolated had been vaccinated against IB with vaccines of the Massachusetts serotype. In vivo protection studies showed that the MA5 vaccine (of the Massachusetts serotype) protected well against challenge with four of these isolates, representing the different serotypes reported in this study.