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Biomedical subjects

M J Studdert

Publications and source records attributed to M J Studdert.

At least 109 records · Page 6Linked to original sources

Restriction endonuclease DNA fingerprinting of respiratory, foetal and perinatal foal isolates of equine herpesvirus type 1.

DNA was prepared from 43 equine herpesvirus type 1 (EHV 1) isolates, 11 of which were from horses with respiratory disease, 22 from aborted equine foetuses, and 10 from foals that died perinatally. The restriction endonuclease DNA fingerprints of 10 of the 11 respiratory isolates, known with certainty to have been recovered from horses with respiratory disease, were entirely different from all but 3 of the 32 foetal or perinatal foal isolates. The exceptional respiratory isolate, EHV 1 Army 183, had a foetal (F) strain fingerprint but this virus cannot be said with certainty to have been isolated from the respiratory tract. The 3 exceptional foetal isolates, had respiratory (R) strain fingerprints, and were recovered from single sporadic abortions. There are no exceptions to the view that only R strains have been isolated from naturally occurring respiratory disease. Also it is clear that major epizootics of abortion (abortion storms) and of perinatal foal mortality are caused by F strains. The data together with an analysis of the epidemiological patterns, particularly in Australia, strongly support the view that F and R strains be regarded as separate species, EHV 1 and 4 respectively.

Abortion, Veterinary↗

Analysis of small and large plaque variants of equine herpesvirus type 3 by restriction endonucleases. Brief report.

Six of 6 equine herpesvirus type 3 (EHV3) isolates, 5 of which were epidemiologically unrelated, produced a mixture of small and large plaque variants in equine foetal kidney cells under methylcellulose. In 4 of 4 instances the cleavage site(s) generating the Bam HI A fragment of large plaque variants was distinct from the site(s) for the same fragment of small plaque variants.

DNA Restriction Enzymes↗

Reconstitution of primary, severe, combined immunodeficiency in man and horse.

Severe combined immunodeficiency disease (SCID) in foals is the only known animal model for the autosomal recessive form of primary SCID in man. A major requirement in the treatment of SCID is the maintenance of the patient in a disease free state until definitive therapy can be undertaken. This paper reviews the current status of prophylactic and definitive therapy in man and the horse. Particular emphasis is placed on the methods of reconstitution available, involving foetal tissues and bone marrow.

Adenosine Deaminase↗

Aspects of the diagnosis, pathogenesis and epidemiology of canine parvovirus.

Between 18 July 1980 and 2 January 1981, 188 samples (145 faeces and 43 intestinal contents) were submitted from dogs with suspected canine parvovirus (CPV) enteritis. CPV was demonstrated in 56 (30%) of these samples; the weekly rate of positive CPV identification was remarkably constant at approximately 30% even though clinical and often post-mortem findings strongly supported a diagnosis of CPV enteritis. The simplest, most sensitive and most rapid method for detection of virus was haemagglutination (HA) which was twice as sensitive as isolation of virus and 8 times as sensitive as electron microscopy (EM). Forty nine of 56 (88%) samples positive for CPV were from dogs less than 1 year old and 44 (79%) CPV-positive samples were from pups less than 6 months old; only one sample from a pup less than 2 months old (pup was 7 weeks old) was positive. An additional 68 samples (53 faeces and 15 intestinal contents) were submitted from Beagle dogs that were part of a colony of approximately 1200 dogs. Epidemiological data pinpoints the entry of CPV into the colony in November 1978 at which time most dogs including pups less than 6 months of age developed antibody to CPV without developing clinical disease. From these data an overview of some aspects of the pathogenesis and epidemiology of CPV is constructed.

Animals↗

Attempted reconstitution of a foal with primary severe combined immunodeficiency.

A foal with primary severe combined immunodeficiency, diagnosed within the first two weeks of life, was maintained with its dam in semi-isolation. The foal received continuous prophylactic antibiotic therapy, plasma from a sibling hyperimmunised with equine adenovirus vaccine, and intensive general nursing care. A full sibling female was selected as a bone marrow donor on the basis of red blood cell cross-matching and mixed lymphocyte reactions. Cyclophosphamide was given before two bone marrow transfusions at 35 and 73 days of age. To prevent graft versus host disease graft versus host disease the foal was maintained on methotrexate therapy. Reconstitution was not achieved nor were there signs of graft versus host disease. The foal died suddenly four days after the second bone marrow transfer when 77 days old. It had remained clinically free of any life threatening infectious disease and at necropsy a remarkable degree of freedom from infectious disease was confirmed. The most notable necropsy findings were bilateral nephrosis and myocardial degeneration and fibrosis. The likely cause of death was an electrolyte imbalance, particularly hypokalaemia, which secondarily affected the myocardium. Renal toxicity caused by the cytotoxic drugs, especially cyclophosphamide, may have contributed to the electrolyte imbalance.

Animals↗

Generalized parvovirus disease in neonatal pups.

Generalized canine parvovirus disease was diagnosed in a litter of pups that died when 3 to 9 days old. Parvovirus was isolated from the heart, lungs, liver, kidneys, and small intestine. Histologically, lesions were characterized by hemorrhage and necrosis in the brain, liver, lungs, kidneys, lymphoid tissues, and gastrointestinal mucosa. Intranuclear inclusions were found in vascular endothelium, heart, lungs, liver, kidneys, adrenal cortex, and gastrointestinal tract.

Animals↗

Optimal conditions for in vitro blastogenesis of feline peripheral blood lymphocytes.

A microculture technique was developed for the in vitro blastogenesis of feline lymphocytes. Blastogenesis of ficoll-diatriazoate gradient separated mononuclear cell, washed blood and whole blood were compared. In general the whole blood cultures yielded higher stimulation indices (SI) than the washed blood or separated mononuclear cell cultures. The effect of several variables on the stimulation of lymphocyte cultures was examined. A cell concentration of 3 x 10(5) cells/well and a 1:20 dilution of washed and unwashed whole blood gave optimal stimulation with concanavalin A (Con A). Phytohaemagglutinin-P (PHA-P) did not give significant levels of stimulation. Inactivated fetal calf serum (FCS) at levels of 2.5% (for washed blood) and 5% (for separated mononuclear cell and whole blood) gave highest SI. Supplementation with FCS was preferable to autologous, homologous or horse sera for all cultures. Optimal SI was obtained in all cultures incubated for 3 days and labelled with 1 microCi tritiated thymidine (3H-TdR) for the last 16 hours. The highest SI were in the range of 70 to 105 (18,764 to 42,681 counts per minute (CPM) for separated mononuclear cell culture, 100 to 165 (28,403 to 45,334 CPM) for washed blood culture and 105 to 186 (41,076 to 69,999 CPM) for whole blood culture.

Animals↗

Immunogenicity of equine herpesvirus type 1 (EHV1) and equine rhinovirus type 1 (ERhV1) following inactivation by betapropiolactone (BPL) and ultraviolet (UV) light.

Some kinetic data on the inactivation of equine herpesvirus type 1 (EHV1) and equine rhinovirus type 1 (ERhV1) by betapropiolactone (BPL) and ultraviolet (UV) irradiation are reported. 0.25% BPL at 37 degrees C for 1 h reduced the titre of EHV1 by greater than 10(3 . 4) and of ERhV1 by greater than 10(4 . 1) TCID50/ml. UV irradiation (334 microW/cm2) produced similar reductions in titre after 2 min. These data were used as a basis for inactivating EHV1 and ERhV1 by the combined action of BPL and UV irradiation. Viruses were exposed to 0.1% BPL for 1 h at 4 degrees C with constant stirring, followed by UV irradiation for 2 min, followed by incubation for 3 h at 37 degrees C. Inactivated EHV1 elicited secondary immune responses only in horses whereas ERhV1 produced primary immune responses in mice (including athymic nu/nu mice), rabbits and probably in horses.

Animals↗

Isolation of an adenovirus antigenically distinct from equine adenovirus type 1 from diarrheic foal feces.

Adenovirus was isolated in equine fetal kidney cell cultures from the feces of 2 foals with diarrhea that also had large numbers (greater than 10(6)/g) of rotavirus particles in their feces. Unlike equine adenovirus type 1 (EAdV1), the fecal EAdV did not hemagglutinate human O, rhesus macaque, or equine RBC. By serum neutralization, the fecal viruses were identical with each other, but showed no relationship to EAdV1. Antiserum prepared against the fecal viruses did not contain hemagglutination-inhibiting antibody to EAdV1. It is proposed that the fecal viruses be considered prototypic of EAdV2. The frequency of neutralizing antibody to EAdV2 in 339 equine serum samples was 77%. Neither EAdV1 nor EAdV2 is related by serum neutralization to any of 30 human adenovirus serotypes.

Adenoviridae↗

In vitro blastogenesis of equine lymphocytes by inactivated equine adenovirus type 1 antigen.

An inactivated equine adenovirus type 1 (EAdV1) vaccine was administered to 4 horses. The horses had virus-neutralizing (VN) antibody titers before they were vaccinated, but developed higher VN antibody titers in response to vaccination. Nonvaccinated control horses did not show increases in VN antibody during the study, indicating that any increase in antibody titer in vaccinated horses was a result of vaccination and not due to an EAdV1 epizootic during the study. Specific EAdV1 in vitro lymphocyte blastogenesis (LB) was evaluated, using lymphocytes from 4 vaccinated and 2 control horses. Horses were vaccinated on days 0 and 14, and the LB assays were conducted on days -4, 0, 3, 7, 10, 14, 17, 21, 24, and 28. Lymphocytes from horses were incubated for 4 days with 2 concentrations of inactivated, concentrated and purified EAdV1 antigen. The LB responses for the 2 control horses showed no significant changes during the study period (maximum stimulation indices to EAdV1 antigen for individual horses were between 2.8 and 3.6). The 4 vaccinated horses showed marked increases in stimulation indices in response to EAdV1 antigen (maximum stimulation indices, between 5.3 and 18.6). In control assays, identical lymphocyte preparations from all horses responded normally to phytohemagglutinin.

Adenoviridae↗

Differentiation of respiratory and abortigenic isolates of equine herpesvirus 1 by restriction endonucleases.

Viruses classified by immunologic criteria as equine herpesvirus 1 cause respiratory disease and abortion in horses. Restriction endonuclease analyses of the DNA's of viruses from animals with respiratory disease and from aborted fetuses show that the patterns for respiratory viruses, while similar to each other, are entirely different from the patterns for fetal viruses. It is therefore proposed that the DNA restriction endonuclease patterns of fetal and respiratory viruses analyzed in this study be designated as prototypic of equine herpesvirus 1 and 4, respectively.

Abortion, Veterinary↗

Isolation of Chlamydia psittaci from cats with conjunctivitis.

Chlamydia psittaci was repeatedly demonstrated in stained smears of conjunctival scrapings from a group of cats in a single household and in 5 instances the organism was isolated by yolk sac inoculation of 6-day-old pathogen free, embryonated hen eggs. Thirteen of 15 cats in the cattery developed conjunctivitis at various times over a 9-month period. The outstanding features of the disease were its severity, chronicity and refractoriness to treatment. Prolonged (2 week) treatment with tetracycline was required to effect clinical recovery. Nine of 14 cats in the household developed significant complement-fixing (CF) antibody titres (greater than 128) to the chlamydia group antigen. A single serum from the owner had a titre of 32 although no associated illness was recognized. Of 134 serums collected from random source cats aged 1 month to 16 years, 17 (12.7%) also contained CF antibody to chlamydia group antigen. This is the first report of the isolation of chlamydia from cats with conjunctivitis outside North America and the first report to indicate general incidence figures for chlamydia infection of cats where vaccination is not used.

Animals↗

Acute and chronic canine parvovirus myocarditis following intrauterine inoculation.

Canine myocarditis virus was inoculated into puppies in utero 8 days before parturition. Puppies were clinically normal at birth. One puppy died 23 days after inoculation and a second puppy was comatose before euthanasia on day 27 post inoculation. Both had acute, non-suppurative myocarditis, and parvovirus was isolated from the heart of each pup. Two other puppies remained clinically normal until euthanasia at 87 and 131 days after inoculation. These latter puppies had extensive, focal fibrosis within the myocardium. Despite weekly monitoring, ECG changes were not recorded. The apparent predilection of canine parvovirus for cardiac myocytes is discussed.

Animals↗

Caliciviridae.

The caliciviruses, as a proposed family Caliciviridae, have a distinct virion morphology with cup-shaped depressions on a spherical capsid surface. The viruses have single-stranded RNA, which has a molecular weight about 2.6 x 10(6) and is infectious. The RNA is covalently linked to a small protein. A single major polypeptide is found in the capsid. A subgenomic RNA, molecular weight about 1 x 10(6), coding for the capsid polypeptide is found in infected cells. Caliciviruses infecting swine, pinnipeds and cats have been characterized. Viruses which are morphologically identical to the known caliciviruses have been identified in human feces; these viruses have been shown to be associated with gastroenteritis, but they have not yet been propagated in the laboratory.

Animals↗