A study of immunoglobulin M antibody to measles, canine distemper, and rinderpest viruses in sera of patients with subacute sclerosing panencephalitis.

Seven boys were studied who had the clinical features of subacute sclerosing panencephalitis (SSPE) and whose brain histology was consistent with SSPE. Measles antigen was detected in the seven brains by the direct fluorescent antibody method. Three out of the seven boys had in their sera measles specific immunoglobulin M (IgM) which was detected by the indirect fluorescent antibody method, and the cell receptors for it were acetone stable. A prozone effect was noted in the sera of two patients. The absorption of one patient's serum with Staphylococcus aureus to reduce the titre of immunoglobulin G (IgG) removed the prozone effect. Two of the boys who had high titres of measles specific IgM also had serum IgM which reacted with canine distemper virus antigen but the titres were eightfold lower. None of the boys had detectable rinderpest specific IgM in their sera.

Changes in circulating immune complex concentrations and antibody titres during treatment of Q fever endocarditis.

Serum samples from 20 patients with Q fever endocarditis were tested for the presence of circulating immune complexes to see whether their concentrations correlated with antibody titres during treatment and whether they could be used to monitor the response to antimicrobial treatment. Circulating immune complexes were found in all 20 patients. The concentrations in 15 patients correlated with either or both of the Q fever phase 1 and phase 2 antibody titres obtained during treatment. In the other five patients no correlation with the antibody titres was found. There was no association between circulating immune complex concentrations and clinical response to treatment.

Prognostic value of Epstein-Barr virus serology in patients with nasopharyngeal carcinoma.

We report the results of a study which examined the prognostic value of Epstein-Barr virus (EBV) serology in patients with nasopharyngeal carcinoma (NPC). Antibody titres to EBV were estimated by indirect immunofluorescence and correlated with the stage of disease. Neither persistent high titres nor falling titres after treatment were found to be reliable indicators of relapse or survival, respectively, in individual patients. By contrast, four-fold rises in titre, particularly of antibodies to the early antigens of the virus were highly significant predictors of relapse. These increases could be seen well in advance of clinical detection of recurrence of the tumour at the primary or metastatic sites.

Serological responses against the pathogenic dimorphic fungus Mucor amphibiorum in populations of platypus (Ornithorhynchus anatinus) with and without ulcerative mycotic dermatitis.

Mucor amphibiorum, a dimorphic fungus, causes ulcerative dermatitis and systemic infections in the platypus Ornithorhynchus anatinus in some river systems in Tasmania but apparently not in other regions of Australia. As yet there are no suitable tests for population surveys, nor for detection of internal lesions in live animals. Consequently, immunoglobulins were purified from the serum of platypuses and anti-immunoglobulin antisera were prepared in rabbits in order to develop an enzyme-linked immunosorbent assay (ELISA) for anti-M. amphibiorum antibodies. Antigens from plate-grown cultures resulted in greater signal-to-noise ratios in indirect ELISA than those from broth-grown cultures. Platypuses with clinical ulcerative dermatitis had elevated anti-Mucor antibody levels compared to apparently unaffected individuals. Seroconversion was observed in one animal coincident with the development of cutaneous ulcers. The results suggested that platypuses in affected rivers were exposed to M. amphibiorum at a higher frequency than the occurrence of clinical disease. Some platypuses from New South Wales had elevated antibody levels but these increased significantly with age suggesting exposure to cross-reactive antigens, although exposure to M. amphibiorum cannot be excluded. Further studies are warranted to determine factors that result in progression from infection to disease, the occurrence of the fungus in areas where disease has not been observed and the specificity of antigen used in ELISA.

Cytochemical assay for differential respiratory activity in roots and root hairs.

The study of the underground parts of plants is often difficult, and as a result roots are often treated as homogeneous physiological entities with respect to root respiration. In this study we demonstrate a partitioning of respiration within root tissues using nitro blue tetrazolium staining and an incident light optical system that permits detailed observations of intact roots. The assay is rapid and easy to perform, and reveals that respiratory activity in roots is not uniform in space and time. The results show that root hairs in particular may be regions of enhanced respiratory activity in some species or in certain developmental or physiological states. This fact has important implications for the role of root hairs in the overall respiratory budget of roots and the energetics of nutrient assimilation. The results suggest that root respiration studies should consider differential respiratory activities of root cell types within roots.

Gross, histological and immunohistochemical features of mucormycosis in the platypus.

Nine male and five female adult free-living platypuses, obtained in a prospective capture-release study from northern Tasmania, exhibited gross features of cutaneous mycosis caused by Mucor amphibiorum. The lesions were present on the hind limbs (six cases), front limbs (four), tail (five), dorsal trunk (three) and ventral trunk (one). They varied in size, and ranged from raised red nodules or plaques, which sometimes exuded purulent material, to ulcerated lesions with central cavitation, red exuding centres and raised epidermal margins. Older lesions were covered either partly or fully by thickened and irregular epidermis. Histological examination of skin biopsies revealed discrete, poorly encapsulated granulomas, or more commonly a diffuse granulomatous or pyogranulomatous inflammation. Inflammatory cells consisted of neutrophils or eosinophils, sparse plasma cells and lymphocytes, many macrophages and occasional multinucleated giant cells. Fibrovascular tissue was diffusely and irregularly scattered in the granulomatous regions. Sphaerules characteristic of M. amphibiorum infection were observed in all lesions. The cutaneous distribution of the lesions and the natural history of the platypus indicated that entry of M. amphibiorum may have been via superficial skin wounds. T cells were the predominant infiltrating lymphoid cells in the diffuse lesions, indicating the importance of the cell-mediated response to infection.

Mumps or Coxsackie A9 virus antibody in the cerebrospinal fluid of patients with meningitis or encephalitis.

Ten patients with meningitis and one patient with encephalitis associated with mumps virus infection were studied and nine patients had mumps virus antibody in their cerebrospinal fluids taken 19-36 days after onset. Another 10 patients with meningitis associated with Coxsackie A9 virus infection were investigated and three patients had significant Coxsackie A9 virus antibody in their cerebrospinal fluids taken 14-20 days after onset. There was evidence that the mumps and Coxsackie A9 virus antibody in the cerebrospinal fluids was produced inside the central nervous system. In the acute and convalescent cerebrospinal fluids there was no correlation between the white blood cell counts and the mumps or Coxsackie A9 virus antibody titres.

Lymphoid neoplasia in the koala.

OBJECTIVE: Correlation of immunophenotype with history, anatomical and morphological features of lymphoid neoplasia in the koala. METHODS: Routine necropsies were performed on 51 koalas with suspected lymphoid neoplasia between 1986 and 1997 in New South Wales and Queensland. Immunophenotyping was by an immunoperoxidase method utilising species cross-reactive antibodies raised against human lymphocytes and an antibody raised against koala IgG. Cases were classified according to organs and tissues affected and the morphological features of neoplastic cells. RESULTS: Twenty-six (51%) of the cases were of the T cell immunophenotype, 12 (24%) were of B cell immunophenotype and 13 (25%) did not stain. The age and sex of koalas did not correlate with immunophenotype (P = 0.686 and P = 1.000, respectively). Thirty-two cases were leukaemic and 36 had multiple organ involvement, probably reflecting presentation of koalas at advanced stages of disease. Abdominal tissue involvement was most common (44 cases), followed by nodal (32), atypical (21) and cervicomediastinal (14). The T cell immunophenotype was over-represented among the leukaemic cases (P = 0.013). Generally, the T cell immunophenotype predominated except for many affected atypical tissues. Neoplastic cells were mostly of medium nuclear size with round to oval nuclei. No correlations were found for cell morphology, mitotic index and immunophenotype. CONCLUSION: The prognostic value of an immunophenotypic, anatomical and morphological basis for the classification of lymphoid neoplasia in the koala currently is limited by the need to detect these neoplasms at an early age, the requirement for freshly fixed tissues and the restricted range of available cross-reacting antibodies.

Diabetes mellitus in a koala (Phascolarctos cinereus).

OBJECTIVE: To describe a case of diabetes mellitus in a koala (Phascolarctos cinereus). DESIGN: A case report with controls. PROCEDURES: We describe clinical and laboratory findings in a 6-year-old, free-living, female koala presented with traumatic injury and subsequently found to have polydipsia, hyperglycaemia and glucosuria. Over a 5 week period, serum biochemical analyses, haematological examinations, urinalyses, measurement of serum insulin and fructosamine concentrations, necropsy, histopathological examination of a range of tissues and immunohistochemical examination of the pancreas for insulin-containing cells were done. For reference purposes, serum insulin and fructosamine concentrations were determined in four and two healthy koalas, respectively, and three healthy koalas' pancreases were examined histologically and immunohistochemically. RESULTS: The koala had persistent hyperglycaemia, hyperlipidaemia, hyponatraemia, hypochloraemia and glucosuria. Serum insulin concentration of the diabetic koala was only marginally smaller than that of healthy koalas, but all concentrations were smaller than reference concentrations in dogs and people. Fructosamine concentration did not allow the diabetic koala to be distinguished from healthy koalas and concentrations of all koala analytes were greater than expected for healthy dogs and people. Histopathological examination revealed extensive degeneration of pancreatic islet cells and fatty infiltration of hepatocytes. Immunoperoxidase staining revealed decreased or absent insulin in the beta calls of the affected koala. CONCLUSION: Clinical signs, clinicopathological results and histopathological changes were consistent with diabetes mellitus. The pathogenesis of the condition could not be determined but may have been related to the administration of a parenteral corticosteroid preparation, the stress of capture or tissue damage and inflammation.

Haematological, serum biochemical and serological features of platypuses with and without mycotic granulomatous dermatitis.

OBJECTIVE: To determine whether there are haematological, serum biochemical and serological differences between platypuses (Ornithorhynchus anatinus) with and without granulomatous dermatitis due to Mucor amphibiorum infection. An additional objective was to establish reference haematological and serum biochemical ranges for the species in Tasmania. DESIGN: A clinicopathological and serological study. ANIMALS: A total of 37 free-living adult platypuses captured from streams and dams in Northern Tasmania were used in the clinicopathological study. Twenty-seven were clinically normal and 10 had mycotic granulomatous dermatitis. A total of 22 platypuses (20 adult and 2 juvenile) were used for the serosurvey. Eighteen were captured from streams in Northern Tasmania, and four were submitted for necropsy. RESULTS: Platypuses with mycotic ulcerative dermatitis had significantly smaller packed red cell volumes, haemoglobin concentrations, lymphocyte counts, serum cholesterol and calcium concentrations, and higher serum globulin and potassium concentrations than clinically normal animals. The lymphopenia and hyperkalaemia were thought to be clinically significant. Numbers of Trypanosoma binneyi in blood smears were similar between the two groups. Diseased platypuses had higher concentrations of serum antibody against Mucor amphibiorum as determined by ELISA compared to clinically normal platypuses. CONCLUSION: Platypuses affected by mycotic granulomatous dermatitis showed haematological and serum biochemical changes when compared to clinically normal animals from the same Tasmanian sites. A serological survey may be a useful method for detecting the prevalence of exposure to Mucor amphibiorum and humoral immunity in platypus populations both in Tasmania and the mainland of Australia.

Hepatitis delta virus infection in Northern Ireland 1970-1989.

The incidence of hepatitis delta virus (HDV) infection in Northern Ireland (1970-1989) was tested by enzyme linked immunosorbent assay in 401 hepatitis B surface antigen (HBsAg) positive sera. Hepatitis delta antigen (HDAg) was tested in 388 patients and antibody to delta antigen (anti-HD) in 401 patients. Four patients (1.03%) were HDAg positive. Nine patients (2.24%) were positive for anti-HD and after acid pre-treatment of sera from eight of these patients, five were positive for HDAg. The overall incidence of HDV markers was 3.27%, which reflects the low incidence in HBsAg carriers in Northern Ireland (who were in high risk groups for delta hepatitis). The use of acid treatment of the sera to break up antigen/antibody complexes has been a useful technological improvement in the identification of this virus.

Echovirus type 7 outbreak in Northern Ireland during 1984.

During 1984, 118 patients with echovirus type 7 infection were diagnosed. The incidence in Northern Ireland was more than three times higher than the rest of the United Kingdom. The outbreak peaked in June and July, with the highest incidence in Belfast and the eastern part of Northern Ireland. All patients were sufficiently unwell to require hospital admission. Aseptic meningitis was the commonest illness (54.2%) followed by gastroenteritis (22.0%), respiratory tract infections (11.9%) and influenza-like illnesses (8.5%). Males (62%) were affected more than females and 50 patients (42%) were less than one year old. The present epidemic had features in common with four previous enterovirus epidemics, except that the under one year age group was predominantly affected and no family or street outbreaks were detected.

An Otofuke-like virus associated with diarrhoea. Case report and electronmicroscopic study.

Virus particles similar to Otofuke virus have been found, together with rotaviruses and astroviruses, by electronmicroscopy in faeces from an infant with diarrhoea in Northern Ireland. Previously Otofuke virus has been found only in Japan whence it may have been carried to this country by a businessman.

Clinical Q fever in Northern Ireland 1962-1989.

Q fever was diagnosed in 443 patients in Northern Ireland between 1962 and 1989. From 1986 onwards there was an increase, which peaked in 1989 with 107 cases of whom 47 were infected in Ballycastle, Co Antrim. There were three outbreaks and 21 clusters of patients with Q fever. Most cases were in April and May which correlated with the peak lambing and calving season. Q fever mainly affected males in the 40-49 year old age group. County Antrim had the highest prevalence rate of 40/100,000 population and also had the most sheep. The number of sheep in Northern Ireland has doubled in the past ten years. Q fever was strongly associated with occupation and animal contact. Eighty-seven patients (19.6%) drank unpasteurized milk. The commonest presenting illnesses were pneumonia (62.8%), influenza-like illness (24.6%), involvement of the heart (9.0%) and hepatitis (1.6%). Thirty-two patients (7.2%) had endocarditis, 20 of whom had prosthetic valves and three of whom died. Coxiella burnetii was present on valves removed from seven patients.

Hepatitis B virus infection in Northern Ireland 1970-1987.

In the 18 years between 1970 and 1987, 504 patients were found to have hepatitis B surface antigen (HBsAg) in their blood. Acute hepatitis was present in 184 patients and six died (3.3%). The annual incidence of acute hepatitis B virus infection in Northern Ireland was about one-quarter that of England and Wales. A decrease in acute infection occurred in 1986-87, while in England and Wales acute infection has fallen by more than half since the peak in 1984. Hepatitis B virus infection in health care staff and patients in high risk groups were reviewed: 32% were in those of foreign origin or who had known foreign contacts. In blood donors there was a marked fall in incidence of hepatitis B surface antigen carriage from 1982 onwards: the incidence in antenatal patients and those screened for rubella antibody (mainly females) was half that of new blood donors in 1972-81. Carrier rates in blood donors and antenatal patients were less than those from other parts of the United Kingdom. All indices show that Northern Ireland has a lower incidence of hepatitis B virus infection than the rest of the United Kingdom.

Human immunodeficiency virus infection in Northern Ireland 1980-1989.

To 31st December 1989, 71 persons are known to have attended medical practitioners in Northern Ireland with a diagnosis of Human Immunodeficiency Virus (HIV) infection. Twenty-one of these persons have had the diagnosis of Acquired Immune Deficiency Syndrome (AIDS) and 11 have died. The distribution of reports in the "at risk" categories of homosexual/bisexual males, injecting drug users, heterosexual males and females was significantly different (p less than 0.001) from those reported in the United Kingdom as a whole. Of tests for HIV infection carried out in patients attending the genitourinary medicine department of the Royal Victoria Hospital between 1987-1989, 0.16% have been positive. The prognostic value of the T4 lymphocyte count at presentation for the subsequent development of AIDS was significant (p = 0.0011). The commonest AIDS indicator disease diagnosed was Pneumocystis carinii pneumonia which was seen in seven of the 21 patients (33%).

Investigation into the characteristics, triggers and mechanism of apnoea and bradycardia in the anaesthetized platypus (Ornithorhynchus anatinus).

Health and conservation research on platypuses (Ornithorhynchus anatinus) may require anaesthesia to reduce stress and the risk of injury to both the animal and the researcher, as well as to facilitate examination and sample collection. Platypus anaesthesia can be difficult to manage, with reports of periods of apnoea and bradycardia described. This study investigated the conditions around sudden-onset apnoea and bradycardia in 163 field-anaesthetized platypuses as part of a health study. Anaesthesia was induced and maintained using isoflurane delivered in oxygen by face mask. Sudden-onset apnoea and bradycardia was observed in 19% of platypuses, occurring either at induction of anaesthesia, during recovery, or both. At induction, occurrence was more often recorded for adults (P = 0.19) and was correlated with low body temperature (P < 0.001), season (P = 0.06; greater incidence in summer) and longer pre-anaesthetic holding time (P = 0.16). At recovery, sudden-onset apnoea and bradycardia occurred only in platypuses that had been placed in dorsal recumbency as part of their examination, and correlated with poor body condition (P = 0.002), time in dorsal recumbency (P = 0.005), adults (P = 0.06), number of fieldworkers (P = 0.06) and females (P = 0.11). The sudden-onset apnoea and bradycardia we observed is likely to result from the irritant nature of isoflurane (stimulating the trigeminal nerve via nasal chemoreceptors). We propose that this mechanism is analogous to that of submersion of the face/nasal cavity in cold water during a natural dive response, but that the term 'nasopharyngeal response' would more appropriately describe the changes observed under isoflurane anaesthesia. Although we did not record any long-term adverse effects on platypuses that had undergone this response, the nasopharyngeal response could complicate the diagnosis of anaesthetic dose-dependent apnoea and bradycardia. Therefore, we suggest that these responses during anaesthesia of platypuses might be avoided by minimizing the stress around capture and handling, as well as reducing the time in dorsal recumbency.

Haemolytic anaemia associated with Theileria sp. in an orphaned platypus.

CASE REPORT: The clinical and laboratory findings in an orphaned juvenile female platypus (Ornithorhynchus anatinus) that presented with a severe anaemia and tick infestation are reported. The animal developed a terminal septicaemia and died. Antemortem clinical pathology, postmortem histopathology and 18S rDNA sequencing supported a diagnosis of extravascular haemolytic anaemia secondary to Theileria ornithorhynchi infection. CONCLUSION: Although T. ornithorhynchi infection is common in the platypus, this is the first case in which it has been shown to cause a haemolytic anaemia in this species and molecular characterisation of the organism has been described. A review of the previous literature concerning T. ornithorhynchi and possible treatment options for future cases are discussed.

Genotypic analysis of Mucor from the platypus in Australia.

Mucor amphibiorum is the only pathogen known to cause significant morbidity and mortality in the free-living platypus (Ornithorhynchus anatinus) in Tasmania. Infection has also been reported in free-ranging cane toads (Bufo marinus) and green tree frogs (Litoria caerulea) from mainland Australia but has not been confirmed in platypuses from the mainland. To date, there has been little genotyping specifically conducted on M. amphibiorum. A collection of 21 Mucor isolates representing isolates from the platypus, frogs and toads, and environmental samples were obtained for genotypic analysis. Internal transcribed spacer (ITS) region sequencing and GenBank comparison confirmed the identity of most of the isolates. Representative isolates from infected platypuses formed a clade containing the reference isolates of M. amphibiorum from the Centraal Bureau voor Schimmelcultures repository. The M. amphibiorum isolates showed a close sequence identity with Mucor indicus and consisted of two haplotypes, differentiated by single nucleotide polymorphisms within the ITS1 and ITS2 regions. With the exception of isolate 96-4049, all isolates from platypuses were in one haplotype. Multilocus fingerprinting via the use of intersimple sequence repeats polymerase chain reaction identified 19 genotypes. Two major clusters were evident: 1) M. amphibiorum and Mucor racemosus; and 2) Mucor circinelloides, Mucor ramosissimus, and Mucor fragilis. Seven M. amphibiorum isolates from platypuses were present in two subclusters, with isolate 96-4053 appearing genetically distinct from all other isolates. Isolates classified as M. circinelloides by sequence analysis formed a separate subcluster, distinct from other Mucor spp. The combination of sequencing and multilocus fingerprinting has the potential to provide the tools for rapid identification of M. amphibiorum. Data presented on the diversity of the pathogen and further work in linking genetic diversity to functional diversity will provide critical information for its management in Tasmanian river systems.