Biomedical subjects

M S McNulty

Publications and source records attributed to M S McNulty.

At least 19 recordsLinked to original sources

PubMed1995

Pathogenesis of porcine circovirus; experimental infections of colostrum deprived piglets and examination of pig foetal material.

The results of virus and antigen distribution following experimental infection of colostrum deprived pigs with pig circovirus (PCV) by oral/nasal and intravenous routes are reported. PCV and antigen were detected using virus isolation and indirect immunofluorescence on cryostat sections respectively. PCV antigen was detected in tissues throughout the body but primarily in spleen thymus, and lung. No PCV antigen or virus was detected in tissue samples from the central nervous system. Examination of pig foetal material from field cases of abortion/stillbirth resulted in 3 PCV isolates from 2 sera and a spleen sample from 2 groups of stillborn piglets from the same farm. No antibody to PCV alone was detected in 160 foetal sera tested. These results suggest that transplacental infection with PCV does occur, possibly prior to foetal immunocompetance. However, it is probably not a significant cause of reproductive disorders in pigs in Northern Ireland.

Animals↗

PubMed1994

Production, preliminary characterisation and applications of monoclonal antibodies to porcine circovirus.

The preparation of monoclonal antibodies (mAbs) to porcine circovirus is described. Preliminary characterisation was carried out on nine mAbs obtained from two fusions and included isotyping, virus neutralisation assays and indirect immunofluorescence staining patterns obtained following immunostaining of both a porcine circovirus (PCV)-persistently infected pig kidney (PK/15/W) and Vero (Vero-PCV) cell line. Significant differences in the staining patterns were observed in both cell lines which appeared to be dependent on the subculture status of the Vero-PCV cultures. The development of a mAb-based antigen capture enzyme linked immunosorbent assay (ELISA) as an aid to virus purification is also described. The use of mAbs for the detection of PCV antigen in cryostat sections from a pig experimentally infected with the virus leading to identification of the sites of replication of PCV is also reported.

Animals↗

PubMed1994

Transcriptional analysis and genome expression of chicken anaemia virus.

Strand-specific riboprobes representative of either strand of the chicken anaemia virus (CAV) replicative form (RF) DNA indicated that only one strand of the RF was transcribed to produce a major 2.0 kb transcript and that the encapsidated DNA strand was of negative sense. Primer extension analysis located a single transcriptional start site at nucleotide position 360 of the CAV sequence. Amplification, cloning and sequencing of the 3' end of the major transcript revealed the polyadenylation site at nucleotide position 21. Northern blot analysis using a series of genomic probes indicated that the 2.0 kb transcript was devoid of splicing and identified a non-transcribed region of the genome. This non-transcribed region was shown to possess promoter activity, enhancing the expression of the human growth hormone reporter gene in a transient gene expression assay. These observations suggest a simple strategy of genome expression involving a functional polycistronic message.

Animals↗

PubMed1994

Some biological and physico-chemical properties of porcine circovirus.

Some important biological and physico-chemical characteristics of porcine circovirus are reported. These include a study on the host distribution in nature, levels of colostrum-derived antibodies in piglets from sero-positive sows and the susceptibility of a range of cell cultures to infection with this virus. The results of haemagglutination studies, resistance to pH 3, chloroform and heat are also reported as are comparative buoyant densities and sedimentation coefficients of porcine circovirus and chicken anaemia virus.

Animals↗

PubMed1992

Cytotoxicity of bovine leucocytes for parainfluenza type-3 virus-infected cells.

The cytotoxic effect of bovine neutrophils, alveolar macrophages, monocytes and lymphocytes for parainfluenza type-3 (PI-3) virus-infected cells in 51chromium-release assays is described. Specific lysis of virus-infected target cells with PI-3 virus antibody and complement was first observed 8 h after infection coincident with the appearance of haemadsorption-positive cells. Specific lysis increased rapidly reaching a peak 18-24 h after infection. This increase was paralleled by the increase in the percentage of cells with surface haemagglutinin. Target cells were subsequently used in 51chromium-release assays between 18 and 20 h after virus infection. Antibody-independent killing of PI-3 virus-infected cells was observed with neutrophils, alveolar macrophages and lymphocytes. Levels of specific lysis up to 30% for neutrophils and 68% for alveolar macrophages were observed, although there was considerable variation in activity from animal to animal. Lymphocyte preparations showed levels of cytotoxicity up to 20% in some cases while monocytes had low killing ability. Addition of PI-3 virus-specific antibodies enhanced killing by neutrophils, monocytes and lymphocytes but inhibited killing by alveolar macrophages. Complement, particularly guinea pig complement, was cytotoxic for virus-infected but not for uninfected cells, and also considerably enhanced the cytotoxic effect of neutrophils and lymphocytes.

Animals↗

PubMed1992

Characterisation of an antigenically unusual virus responsible for two outbreaks of Newcastle disease in the Republic of Ireland in 1990.

Antigenic characterisation of two highly virulent virus isolates from outbreaks of Newcastle disease on two closely connected farms in County Monaghan, Republic of Ireland, in 1990 showed the viruses to be indistinguishable but unlike other Newcastle disease viruses so far tested. However, they appeared to be antigenically closest to avirulent viruses isolated from waterfowl from several countries and from chickens in Northern Ireland in 1986. Despite the antigenic differences, chickens vaccinated with a live commercial Hitchner B1 vaccine were protected against intramuscular challenge with one of the virulent isolates.

Animals↗

PubMed1992

Effect of "in vitro" exposure of bovine alveolar macrophages to different strains of bovine respiratory syncytial virus.

A vaccine strain of respiratory syncytial (RS) virus and an isolate from pneumonic calves (AC2) were inoculated onto cultures of bovine alveolar macrophages recovered by lung lavage, and the functional properties of the cells observed over a period of 10 days. In most cultures no infectious virus was produced although immunofluorescence indicated the presence of virus antigens in some cells. No significant difference was noted between infected and control macrophage cultures in their capacity to phagocytose latex particles (neutral phagocytosis), although the ability to phagocytose complement-coated Candida krusei cells was affected, particularly with the AC2 strain after 6 days. Killing of C. krusei cells was slightly affected by infection of macrophages with the vaccine strain and was dramatically affected by infection with strain AC2. C3b and Fc receptor expression was adversely affected by both virus strains. Production of neutrophil chemotactic factors was increased in cultures infected with both strains, but was greater with AC2, suggesting that some properties of the cells were activated.

Animals↗

PubMed1992

Characterization of viral DNAs from cells infected with chicken anaemia agent: sequence analysis of the cloned replicative form and transfection capabilities of cloned genome fragments.

The viral DNAs induced by the unclassified animal virus, chicken anaemia agent (CAA), during replication in MDCC-MSB 1 cells have been investigated. Analyses after S1 nuclease, restriction endonuclease and denaturation treatments indicated that infected cell extracts contained genome-size, single-stranded DNA (M(r) 2.3 kb), closed and open circular, double-stranded replicative form (RF) DNAs (M(r) 2.3 kbp) and a population of smaller double-stranded DNAs (M(r) 0.8 kbp). Recombinant plasmids containing 2.3 kbp CAA RF fragments cloned at the PstI, BamHI and EcoRI sites failed to transfect MDCC-MSB 1 cells. However, one plasmid, which contained two 2.3 kbp CAA RF fragments ligated in tandem at the PstI site, and cloned 2.3 kbp PstI, BamHI and EcoRI fragments, excised from their respective plasmids by restriction endonuclease digestion, were capable of transfection. The nucleotide sequence of the circular genome (2298 bp) of the Cux-1 isolate of CAA has indicated the presence of three overlapping open reading frames (ORFs) of 52 kDa, 24 kDa and 13 kDa on one strand. The existence of these ORFs was corroborated by analyses of partial sequences from three other isolates. The non-coding region of the CAA genome contained sequences with putative regulatory function. These results are discussed in relation to the "rolling circle" model of DNA replication.

Amino Acid Sequence↗

PubMed1992

Immuno-gold labelling of turkey rhinotracheitis virus.

The morphology and morphogenesis of five viruses isolated in Great Britain, France and South Africa from turkeys with rhinotracheitis were examined. The five isolates were antigenically related by immunofluorescence and were indistinguishable by negative contrast and thin section electron microscopy. Negative contrast electron microscopy of infected Vero cell cultures revealed ortho- or paramyxovirus-like particles and helical nucleocapsids 14 nm in diameter with a pitch of 6 nm. The viral nature of these structures was confirmed by immuno-gold labelling, using a hyperimmune rabbit antiserum to TRT virus and 15 nm gold-labelled goat anti-rabbit IgG. Ultrastructural changes characteristics of paramyxovirus infection were observed in Vero cell cultures infected with each of the five TRT virus isolates. These alterations, which included areas of localised thickening of plasma membrane, associated cytoplasmic inclusions of nucleocapsids and budding virus particles also labelled specifically following immunogold staining. These observations are in accord with the suggestion that TRT virus is an avian pneumovirus.

Animals↗

PubMed1992

Detection and differentiation of chicken anemia virus isolates by using the polymerase chain reaction.

Complementary oligonucleotide primers which flank a 675-bp DNA fragment encompassing part of the putative gene for the capsid protein of chicken anemia virus (CAV) were used for the enzymatic amplification of CAV DNA by the polymerase chain reaction (PCR). Application of a dot blot hybridization assay by using a 32P-labeled cloned CAV DNA probe allowed PCR product amplified from as little as 0.1 fg of the target DNA sequence to be detected. When it was used for PCR amplification, DNA extracted from thymus tissue by a guanidine isothiocyanate-based method proved to be more efficient than that extracted by methods involving phenol or boiling. DNAs specified by 14 CAV isolates originating in the United Kingdom, Ireland, Germany, Sweden, the United States, Japan, and Australia were amplified. Restriction endonuclease analysis of the PCR-amplified DNAs with the enzymes HaeIII, HinfI, and HpaII indicated that the 14 CAV isolates can be assigned to seven groups, with isolates from different countries usually exhibiting the greatest number of restriction site differences.

Anemia↗

PubMed1992

Effect of macrophages and in vitro infection with parainfluenza type 3 and respiratory syncytial viruses on the mitogenic response of bovine lymphocytes.

Bovine blood lymphocytes, depleted of macrophages by absorption on plasma-gelatin coated plastic flasks, followed by passage through Sephadex G-10 columns, failed to respond to pokeweed mitogen stimulation. Adherent monocytes or alveolar macrophages added to purified lymphocyte preparations at 10% or less were able to restore the transformation response. Exposure of alveolar macrophages or purified lymphocytes to 2 bovine respiratory syncytial virus strains for 24 hours substantially reduced the transformation response when mixed with uninfected lymphocytes or macrophages. Exposure of alveolar macrophages or purified lymphocytes to 2 bovine parainfluenza type 3 virus strains produced a similar reduction in activity after 48 hours. Heat inactivation of the viruses removed their inhibitory ability. Immunofluorescence studies revealed that both alveolar macrophages and lymphocytes were permissive for parainfluenza type 3 virus, whereas only a small number of alveolar macrophages and lymphocytes were infected with respiratory syncytial virus. The results suggest that both viruses are capable of adversely affecting the interaction between macrophages and lymphocytes, although the mechanisms by which this is achieved may be different.

Animals↗

PubMed1992

Effects of two adenoviruses (type 1 and type 8) on functional properties of bovine alveolar macrophages in vitro.

Cultures of bovine alveolar macrophages were inoculated with type-1 and type-8 adenoviruses, initially isolated from calves with respiratory tract disease, and functional properties of the cells were observed over a period of 10 to 11 days. Both viruses replicated in macrophages; viral titers were low (less than 3.75 log10 TCID50/0.1 ml), and intranuclear inclusions were detected by indirect immunofluorescence in 5 to 10% of the cells from 3 days after inoculation. Highest titers were induced by type-1 adenovirus, which also induced the greatest functional changes. Expression of Fc and complement receptors was reduced by both viruses, although the greatest effects were seen with type 1. Phagocytosis of Candida krusei cells was reduced following type 1 infection, whereas phagocytosis in type-8-infected cells was not different from that of noninfected macrophages. Ability to kill ingested Candida cells also was reduced following type-1 infection, whereas type-8-infected macrophages had lower killing ability only at 2 to 4 days after inoculation. Neither virus had substantial effects on the production of neutrophil chemotactic factors by the macrophages.

Adenoviridae↗

PubMed1991

Comparison of three animal viruses with circular single-stranded DNA genomes.

No common antigenic determinants and no DNA sequence homologies were detected when three animal viruses, chicken anaemia agent (CAA), porcine circovirus (PCV), and psittacine beak and feather disease virus (PBFDV), all of which possess circular single-stranded DNA genomes, were compared. Negative contrast electron microscopy showed that PCV and PBFDV particles were 30% smaller than CAA particles and lacked the surface structure of CAA.

Animals↗

PubMed1991

Biological characterisation of Australian isolates of chicken anaemia agent.

Three Australian isolates of chicken anaemia agent (CAA) resisted treatment at 70 degrees C for 5 min and chloroform treatment. Although minor antigenic differences were detected using monoclonal antibodies to CAA, the Australian isolates were indistinguishable from the reference Cux-1 and Gifu-1 isolates in cross-immunofluorescence and cross-neutralisation tests employing polyclonal chicken antiserums. The Australian viruses were pathogenic for intramuscularly inoculated 1-day-old SPF chicks, but were less pathogenic for 7-day-old chicks. Thus the Australian isolates of CAA did not differ significantly in these properties from previously characterised CAA isolates from other continents.

Anemia↗

PubMed1991

Dot blot hybridization assay for chicken anemia agent using a cloned DNA probe.

A dot blot hybridization assay capable of detecting chicken anemia agent (CAA)-specific DNA in tissues from infected birds has been developed. The assay uses a 32P-labeled DNA probe prepared from cloned CAA-specific fragments representing the entire virus genome and has a sensitivity limit of between and 1 and 10 pg. DNAs from CAA isolates originating in the Federal Republic of Germany, Japan, the United States, the United Kingdom, and Australia were detected. Investigation of specimens from experimentally infected chicks indicated that virus-specific DNA was detected in the tissues of birds from 5 through 42 days after infection and that greater amounts were usually detected in the thymus than in the spleen, liver, feces, or blood. Tissues from specific-pathogen-free and broiler chicks which had become infected at an older age through contact with experimentally infected anemic chicks also contained CAA-specific DNA detectable by the assay. Thymuses from 1- to 2-week-old chicks from eight commercial broiler flocks which had been showing clinical signs characteristic of anemia-dermatitis syndrome were found positive by the hybridization technique, but thymuses from chicks obtained from broiler flocks which did not show such signs were found negative. Of the 35 positive samples (from 46 samples tested), 19 (54%) contained virus-specific DNA in sufficiently great amounts to permit 4-h autoradiography exposures and sample throughput times of 2 days. When compared with virus isolation, the CAA dot blot hybridization assay is time- and labor-saving.

Anemia↗

PubMed1991

Ultrastructural features of alveolar lesions in induced respiratory syncytial virus pneumonia of calves.

Ultrastructural changes occurred in alveolar epithelium in the acute and repair stages of induced respiratory syncytial virus pneumonia induced in eight calves (calf Nos. 1-7, 3 to 6 days old and calf No. 8, 2 weeks old), using a bovine strain of respiratory syncytial virus. Five of the calves were Friesians, three were Hereford x Friesians, and all were male. Tissues from three mock-infected control calves (two Friesian, one Hereford x Friesian) were also examined. Evidence of respiratory syncytial virus infection was observed in both type I and type II pneumocytes from day 4 to day 8 after infection. Infection of type I pneumocytes frequently resulted in necrosis. The response of type II pneumocytes to respiratory syncytial virus infection varied and included hypertrophy, hyperplasia, and syncytial formation. In some infected type II pneumocytes, there were numerous irregular projections of the cell surface, associated with viral budding. Hypertrophy and hyperplasia of type II pneumocytes, epithelial syncytium formation, and irregular cytoplasmic projections from epithelial cells caused considerable thickening of respiratory membrane and occlusion of alveolar lumina. Neutrophils were frequently observed in close association with virus-infected epithelial cells, but evidence of respiratory syncytial virus infection and replication was not observed in alveolar macrophages or neutrophils. Proliferation of type II pneumocytes appeared to play a major role in maintaining the integrity of the alveolar epithelium during the acute stage of the experimental pneumonia. Increased numbers of type II pneumocytes were present on alveolar walls, particularly from 4 to 8 days after infection, and some alveoli were lined entirely by this cell type. In some areas, however, squamous epithelial cells were also involved in covering exposed alveolar basement membrane.

Animals↗

PubMed1991

Ultrastructural features of lesions in bronchiolar epithelium in induced respiratory syncytial virus pneumonia of calves.

Ultrastructural changes were observed in bronchioles in acute and repair stages of respiratory syncytial virus pneumonia induced in eight young calves (calf Nos. 1-8) using a bovine strain of respiratory syncytial virus. Five of the calves were Friesians and three were Hereford x Friesians and all were male. Tissues from three mock-infected control calves (two Friesian, one Hereford x Friesian) were also examined. Calves were from 3 to 6 days old at the time of first inoculation, with the exception of calf No. 8, which was 2 weeks old. In the acute stage of the induced pneumonia, evidence of respiratory syncytial virus replication and release was demonstrable in both ciliated and non-ciliated bronchiolar epithelial cells, with the virus-releasing process most obvious at 4 and 5 days after infection. Respiratory syncytial virus infection of bronchiolar epithelium was associated with various changes, including hypertrophy, hyperplasia, and formation of syncytia. Necrosis of epithelial cell structures usually appeared to be preceded by their desquamation from bronchiolar walls. Respiratory syncytial virus infection resulted in considerable damage to the bronchiolar ciliary apparatus. Such damage was seen as early as 1 day post-infection and was still obvious at 10 days post-infection. Neutrophils were closely associated with respiratory syncytial virus infected epithelial cells and evidence of neutrophil fusion with infected epithelial cells was seen. These observations suggest that neutrophils may be involved in killing respiratory syncytial virus infected cells and that neutrophils might play an important role in early antiviral defense against respiratory syncytial virus at a time when antibody levels are low and other cellular defenses are not fully in play. Bronchiolar repair was evident from 6 days after infection and was well advanced at 10 and 13 days after infection.

Aged↗

PubMed1990

Purification and biochemical characterization of chicken anaemia agent.

Chicken anaemia agent (CAA) was purified using differential centrifugation and successive cycles of equilibrium density gradient centrifugation using sucrose and CsCl. The purification method was dependent on the use of an antigen-detecting ELISA based on a CAA-specific monoclonal antibody. Virus particles banded at a density of 1.33 to 1.34 g/ml in CsCl and measured 23.5 +/- 0.8 nm in diameter. Purified preparations contained one major polypeptide (Mr 50,000) and a single-stranded, circular DNA (2.3 kb). CAA shares some of the biochemical characteristics possessed by porcine circovirus and the virus associated with psittacine beak and feather disease.

Animals↗