Chlamydiosis in captive white-winged doves (Zenaida asiatica).

Chlamydia psittaci was isolated from the spleen of a moribund white-winged dove (Zenaida asiatica). The isolate was serotyped as the serovar B that is commonly isolated from pigeons. A fourfold increase in the titer of antichlamydial IgM activity occurred in that bird in paired serum samples tested by chlamydial elementary body agglutination (EBA) and a greater than or equal to fourfold decrease of IgG occurred by direct complement fixation (DCF). The increases or decreases of EBA and DCF titers in other clinically ill birds that were treated with tetracycline varied, as normally occurs in cases of avian chlamydiosis. Titers in clinically normal birds were consistent with past infections. These birds were from a captive group of about 200 birds to be used for breeding and reproduction research. A small sample of recently caught wild birds was serologically negative for chlamydial antibody activity.

Analysis of a hypervariable region in the 3' non-coding end of the infectious bronchitis virus genome.

Previous studies on infectious bronchitis virus (IBV) cDNA have identified a region of about 184 bases in the 3' non-coding terminus of both the U.S. prototype strain (Beaudette) and a Japanese strain (KB8523), that was not present in an antigenically closely related U.S. strain, Massachusetts (Mass) 41 (Boursnell et al., 1985; Sutou et al., 1988). In order to investigate the origin and function of this region and its occurrence in nature, the cDNA sequences of the 3' non-coding regions of three additional strains of IBV, Gray, Arkansas (Ark) 99 and Holland (Holl) 52, were determined and compared to the sequences of the Beaudette, KB8523 and Mass41 strains. Not only was this Urich sequence absent from the 3' non-coding region of the Mass41 strain, it was also highly variable, especially in comparison to the highly conserved 3' non coding region downstream of this sequence. Computer analyses of the sequences adjacent to this hypervariable region (HVR) showed that the 3' end of the IBV genome was highly conserved downstream of this region, with 94.3 to 97.8% similarity. However, the similarities for the HVR ranged from 53.2% between Holl52 and Ark99, to 92.8% between Beaudette and Gray. The flanking sequences were not only conserved but these sequences upstream and downstream of the HVR also formed mirrored images.

Comparative analyses of the nucleocapsid genes of several strains of infectious bronchitis virus and other coronaviruses.

The natural sequence variations of the nucleocapsid genes of the Gray, Arkansas99 (Ark99), and Holland52 (Holl52) strains of infectious bronchitis virus (IBV) were determined. These were compared with previously published sequencing data of other IBV strains, as well as other coronaviruses, in order to correlate the serological and evolutionary relationship of coronaviruses. IBV nucleotide sequence alignment shows that overall the sequences are highly conserved, with homologies from 91.1 to 96.5%. However, there are also two regions (730 to 800 and 1138 to 1166) that appear to be even more highly conserved. Overall, the nucleocapsid protein is highly variable both in size and composition between coronavirus major antigenic groups but is conserved within these groups. A phylogenetic tree of the nucleocapsid protein of various coronaviruses indicates that the coronaviruses fall into distinct groups that correspond to the three major antigenic groups; however, a phylogenetic tree of the IBV nucleocapsid shows that this does not hold true for the type specific antigenic groups of IBV.

Detection of avian infectious bronchitis viral infection using in situ hybridization and recombinant DNA.

A recombinant DNA probe with specificity for the 3' end of genomic RNA from the Ark 99 strain of infectious bronchitis virus (IBV) was found to hybridize with extracted RNA of three strains with the Ark serotype, as well as the Mass41, Holl52, Gray, JMK, Conn, Fla and SE17 strains of IBV. Viral infection was detected in the cytoplasm of chicken embryo kidney cells inoculated with Mass41, Ark99, SE17 or two recent field isolates of IBV using in situ cytohybridization and a biotinylated probe. In vivo infections were detected in individual cells of tracheas and lungs 2,4, and 6 days after inoculation of chicks with Mass41 and Ark99. In situ hybridization of Ark99 infected tissue sections using 32P-dATP labelled probe indicated that more viral replication was present in the trachea on day 4 than either days 2 or 6; whereas more viral RNA was found in the lungs on day 6 than days 2 or 4 after inoculation.

Comparisons of the structural proteins of avian infectious bronchitis virus as determined by western blot analysis.

The antigenic diversity of ten strains of avian infectious bronchitis virus (IBV) was examined by Western blot analyses using polyclonal antisera specific for the Massachusetts 41 (M41), Gray, Arkansas DPI (Ark DPI), Connecticut (Conn) and Australian T (Aust T) serotypes. Although antigenic variation was found in all three structural viral proteins, the matrix protein appeared to be antigenically the most highly variable. Four distinct antigenic groups, which did not correspond to virulence or pathotype, could be defined according to the variations observed in the matrix protein. Somewhat less variation was seen in the spike polypeptide. The only variation in the nucleocapsid protein was indicated by the lack of a detectable reaction between the Aust T antiserum and the Ark DPI nucleocapsid protein. Antisera made against M41 had the broadest reactivity while antisera against Aust T, the only strain tested which was exotic to the U.S.A., had the greatest specificity.

Pasteurella haemolytica bacteriophage: identification, partial characterization, and relationship of temperate bacteriophages from isolates of Pasteurella haemolytica (biotype A, serotype 1).

Pasteurella haemolytica (biotype A, serotype 1) isolates (n = 15) from the upper respiratory tract of clinically normal cattle, as well as from lung lesions from cases of fatal bovine pasteurellosis, were examined for the presence of bacteriophage after irradiation with UV light. Treatment of all P haemolytica isolates with UV irradiation resulted in lysis of bacteria due to the induction of vegetative development of bacteriophages. The extent of growth inhibition and bacterial lysis in irradiated cultures was UV dose-dependent. Bacterial cultures exposed to UV light for 20 s reached peak culture density between 60 and 70 minutes after irradiation; thereafter, culture density declined rapidly, so that by 120 minutes, it was approximately 60% of the original value. When examined ultrastructurally, lytic cultures from each isolate revealed bacteriophages with an overall length of approximately 200 nm and that appeared to have a head with icosahedral symmetry and a contractile tail. Cell-free filtrate from each noninduced bacterial isolate was inoculated onto the other bacterial isolates in a cross-culture sensitivity assay for the presence of phages lytic for the host bacterial isolates. Zones of lysis (plaques) did not develop when bacterial lawns grown from the different isolates were inoculated with filtrates from the heterologous isolates.