Sub-cellular localisation of alkaline phosphatase activity in the cytoplasm of tammar wallaby (Macropus eugenii) neutrophils and eosinophils.

Alkaline phosphatase (ALP) has been used in studies of neutrophil morphology and function as a marker for identifying different granule populations. In human neutrophils, ALP is found within secretory vesicles, a rapidly mobilisable vesicle population important for upregulating membrane receptors during early activation. Intra-cellular ALP activity in the heterophils of rabbits and guinea pigs, in contrast, is found only in secondary granules. The neutrophils and eosinophils of tammar wallabies (Macropus eugenii) have previously been reported to contain large amounts of ALP activity when stained using routine cytochemical techniques. To define the subcellular location of ALP in this species, cell suspensions were examined using cerium chloride cytochemistry and transmission electron microscopy (TEM). ALP was found in 2 distinct cytoplasmic compartments. One compartment displayed morphology consistent with a subpopulation of secondary granules while a second tubulo-vesicular population appeared similar to the secretory vesicles of human neutrophils. Thin tubular vesicles containing ALP were also identified within the cytoplasm of tammar wallaby eosinophils. Large numbers of ALP-containing vesicles have not been recognised previously in eosinophils and this may represent a novel cytoplasmic compartment. In both cell types, ALP-containing structures showed alteration in morphology following stimulation with N-formyl-Met-Leu-Phe (fMLP) and PMA.

Use of real-time quantitative polymerase chain reaction to monitor antibiotic therapy in a dog with naturally acquired Mycoplasma haemocanis infection.

Mycoplasma haemocanis is a hemotropic bacterium that can be associated with acute hemolytic disease in immunocompromised or splenectomized dogs. The present case report describes for the first time the use of real-time quantitative polymerase chain reaction (qPCR) to monitor M. haemocanis infection in a splenectomized dog. The report also describes the application of real-time qPCR for the analysis of deoxyribonucleic acid extracted from stained blood films. The analysis of blood films from the time of initial presentation allowed a retrospective confirmation of M. haemocanis infection. The M. haemocanis copy numbers remained high throughout antibiotic treatment of this dog. A decline in copy numbers was only recorded after 11 months of therapy, when improvements in clinical and hematological indices were also noted. Clearance of infection was not achieved, and the dog remained positive for M. haemocanis at 3.5 months postcessation of antibiotic therapy. Cytological examination of blood films for the presence of organisms was insensitive for the detection of parasitemia.

An evaluation of the performance of OraQuick ADVANCE Rapid HIV-1/2 Test in a high-risk population attending genitourinary medicine clinics in East London, UK.

To date, no data have been published on the use of OraQuick ADVANCE Rapid HIV-1/2 Test (OraQuick) in the UK. We report preliminary findings of an ongoing evaluation of OraQuick in UK genitourinary (GU) medicine clinics. A total of 820 samples from patients in high-risk groups for HIV were tested with OraQuick and results were compared with standard HIV antibody testing. HIV prevalence (enzyme immunoassay [EIA]) was 5.73%, sensitivity of OraQuick was 93.64% (95% CI 82.46-98.66%), specificity 99.87% (99.28-100%), positive predictive value 97.78% (88.27-99.94%) and negative predictive value 99.61% (98.87-99.92%). This includes three false-negatives considered to be due to observer error and now rectified by further training. This has increased test sensitivity to 100%. Our observed test performance of OraQuick compares well with EIA and with other rapid tests. We believe that simple, non-invasive antibody detection tests such as OraQuick can increase HIV testing and diagnosis in UK GU medicine and community settings.

Hematologic characteristics of captive western barred bandicoots (Perameles bougainville) from Western Australia.

BACKGROUND: The western barred bandicoot (Perameles bougainville) is an Australian marsupial species now considered endangered as a consequence of habitat destruction and predation. A recently discovered papillomatosis syndrome is hindering efforts to repopulate this species. Hematology reference intervals have been lacking for P bougainville, preventing optimal interpretation of hematology results from wart-affected and clinically normal animals. OBJECTIVES: The purpose of this study was to establish hematology reference values and describe morphologic characteristics of blood cells of healthy western barred bandicoots. METHODS: Fifty-nine whole blood samples were collected by jugular venipuncture into EDTA from 47 clinically healthy captive western barred bandicoots at 3 locations on the Western Australian mainland. A CBC was performed using an ADVIA-120 analyzer. Data were compared on the basis of geographic location, sex, age, and lactation status, and reference intervals were calculated. Blood cell morphology was evaluated using light microscopy, and transmission and scanning electron microscopy. RESULTS: Significant differences were found based on sex (RBC indices, fibrinogen), age (% polychromatophilic RBCs), and geographic location (RBC, neutrophil, and lymphocyte counts, MCHC, % polychromatophilic RBCs, fibrinogen). Combined reference intervals were calculated for hemoglobin concentration (122-165 g/L), HCT (0.36-0.49 L/L), and total WBC (2.9-14.9 x 10(9)/L), monocyte (0-0.6 x 10(9)/L), eosinophil (0-0.9 x 10(9)/L), and total plasma protein (47-63 g/L) concentrations. Leukocyte, erythrocyte, and platelet morphology were similar to those of other marsupial peramelid species. Nuclei in neutrophils, monocytes, and eosinophils occasionally had an annular configuration. CONCLUSIONS: Reference intervals and blood cell morphology obtained in this study will be useful for the evaluation of laboratory data from ill animals and assist with population health monitoring of western barred bandicoots.

Effects of temperature and duration of sample storage on the haematological characteristics of western grey kangaroos (Macropus fuliginosus).

OBJECTIVE: To investigate the effects of storage duration and temperature on haematological analyses performed on blood from the western grey kangaroo (Macropus fuliginosis). METHOD: Blood samples from five western grey kangaroos were stored at 4 degrees C, 24 degrees C and 36 degrees C. Each sample was analysed haematologically over a 5-day period. RESULTS: The blood samples maintained optimal stability at 4 degrees C. At this temperature the haematological values remained essentially unchanged for the duration of the study, while samples stored at 36 degrees C and 24 degrees C showed significant changes in some haematological measures by 12 h and 48 h, respectively. Disturbances in leukocyte morphology were evident, to varying degrees, in all samples. CONCLUSIONS: Blood samples from macropodids should be tested within 48 h of collection if stored at a room temperature of about 24 degrees C. Where testing is to be delayed for more than 48 h, samples should be refrigerated as soon as possible. Exposure of samples to heat in excess of 24 degrees C should be avoided at all times.

The validation of self-reported smoking status by analysing cotinine levels in stimulated and unstimulated saliva, serum and urine.

OBJECTIVES: Cotinine, a nicotine metabolite, can be used to measure exposure to tobacco smoke. The aim of this study was to compare cotinine levels in different biological fluids collected from both smokers and non-smokers and to relate the findings to self-reported smoking status. Data were also collected concerning the acceptability of the differing methods of sample collection. MATERIAL AND METHOD: Patients recruited to the study were asked to provide samples of urine, blood and saliva (both stimulated and unstimulated). Data collected from patients by questionnaire included information on smoking behaviour such as daily number of cigarettes and environmental exposure to smoke. After the sample collection, patients were asked to rate the acceptability of each sampling method. Samples were analysed using enzyme immunoassay (EIA) kits. RESULTS: In total, 80 patients participated, with 49 being smokers and 31 being non-smokers. There was clear differentiation between smokers and non-smokers (P < 0.001) for all the different samples in terms of cotinine. A significant relationship was seen between cotinine and daily number of cigarettes for both salivas and urine (all P < 0.001) but not for serum. Participants found serum and urine collection methodologies 'very acceptable' (67 and 66%, respectively) whereas 9% found collection of stimulated saliva 'not at all acceptable'. CONCLUSION: Cotinine, whatever the collection method and analysed by EIA kits, shows good differentiation between smokers and non-smokers. Salivary samples have the advantage of being non-invasive, although collection methodology is important, as cotinine levels may vary.

A new motor-driven surgical probe and its in vitro comparison with the Cavitron Ultrasonic Surgical Aspirator.

The Cavitron Ultrasonic Surgical Aspirator (CUSA) has been applied in neurosurgery for several years, but its mode of action is not yet clear and its efficiency at removing soft tissue has not been quantified. We describe here how we have measured the rate of removing soft tissue per unit time, taking ox-liver tissue as the test material. A motor-driven vibrator/aspirator has been developed in our laboratory. It has permitted us to examine the effect of varying independently frequency, amplitude of the vibration, and suction pressure on the removal rate. The results of this investigation show that beyond a certain tip acceleration amplitude (about 100 g) the removal rate does not increase significantly. Also the removal rate is more or less independent of vibration amplitude within the range between 300 micron and 1 mm. Our in vitro experiments with the new probe show that a tip acceleration of about 100 g is enough to remove ox-liver tissue and then the rate of removal is comparable to that obtained with the CUSA operating at maximum vibration amplitude. Analysis of the particle size of the debris collected from CUSA and from our motor-driven device shows that the particle size distribution is similar over the range of 0.5 micron less than d less than 250 micron.