Kostas Papasouliotis

Description of outcomes of experimental infection with feline haemoplasmas: Copy numbers, haematology, Coombs' testing and blood glucose concentrations

The aim of this study was to compare blood copy, haematological and glucose values between cats experimentally infected with either Mycoplasma haemofelis (Group HF: 10 cats), ‘Candidatus M. haemominutum’ (Group HM: 3 cats) or ‘Candidatus M. turicensis’ (Group TU: 3 cats). Blood samples were collected regularly up to 85 days post-infection (DPI) for haemoplasma real-time quantitative PCR, haematology, Coombs’ testing and blood glucose measurement. Statistical analysis was performed using a general linear model (ANOVA) appropriate for a repeated measures experiment with significance set as P < 0.05. Cats in Group TU had significantly lower blood copy numbers than cats in Group HF (P < 0.001) and HM (P < 0.001). All Group HF cats developed anaemia (often severe), macrocytosis and evidence of erythrocyte-bound antibodies whereas Groups HM and TU cats did not. Group HF had significantly lower PCVs, haemoglobin concentrations and red blood cell counts, and significantly higher mean cell volumes, than Groups HM and TU. In Group HF, erythrocyte-bound antibodies reactive at 4 °C (both IgM and IgG) appeared between 8 and 22 DPI and persisted for two to four weeks, whereas those reactive at 37 °C (primarily IgG) appeared between 22 and 29 DPI and persisted for one to five weeks. In most cats antibodies appeared after the fall in haemoglobin started. Although Group TU had significantly lower glucose concentrations than Groups HF (P = 0.006) and HM (P = 0.027), mean blood glucose concentrations remained within the reference range in all groups. This study demonstrates that M. haemofelis infection, in contrast to ‘Candidatus M. haemominutum’ and ‘Candidatus M. turicensis’ infection, can result in a severe macrocytic anaemia and the development of cold and warm reactive erythrocyte-bound antibodies.

Serum protein electrophoresis in 155 cats

All serum protein electrophoresis (SPE) results obtained between 2002 and 2009 from clinical cases presented to the University of Bristol Feline Centre were examined retrospectively. One hundred and fifty-five results met the inclusion criteria. Signalment and final diagnoses were obtained from the case records. Clinical cases were classified as having normal or abnormal SPE results by comparison to reference intervals for SPE created using 77 clinically normal cats. Abnormal results were then further divided according to the specific SPE abnormality. Cases were also categorised, according to the final diagnosis, using the DAMNITV classification system. Of the 155 cases, 136 (87.7%) had abnormal SPE results, most commonly due to a polyclonal increase in gamma globulins. A monoclonal gammopathy occurred in four cats; one with feline infectious peritonitis (FIP), one with lymphoma and two cases of splenic plasmacytoma (one suspected, one confirmed). The most common DAMNITV classification associated with SPE abnormalities was infectious/inflammatory disease (80/136; 58.8%), including 39 cats diagnosed with FIP.

Prevalence of Potentially Pathogenic Enteric Organisms in Clinically Healthy Kittens in the UK

Faecal samples were collected from 57 clinically healthy kittens presented for initial vaccination, in the UK. Routine bacteriological examination identified Salmonella species in one and Campylobacter species in five samples. Polymerase chain reaction (PCR) detected the presence of Campylobacter species in a further four samples. Routine parasitological examination revealed Toxocara species ova in nine (including four kittens stated to have been administered an anthelmintic) and Isospora species in four samples. No Giardia or Cryptosporidium species were detected by routine methods. A Giardia species enzyme-linked immunosorbent assay (ELISA) test kit designed for use in cats was positive in three kittens. A similar test kit designed for use in humans was negative in all samples and produced negative results even when known positive samples were tested. Potentially pathogenic enteric organisms were detected in 19 kittens by routine methods and 26 (prevalence 45%) by all methods. The high prevalence in asymptomatic kittens highlights the possibility that the detection of these organisms in kittens with gastrointestinal disease may be an incidental finding.

A comparison of orocaecal transit times assessed by the breath hydrogen test and the sulphasalazine/sulphapyridine method in healthy beagle dogs

Orocaecal transit time (OCTT) was assessed in six healthy beagles by means of the breath hydrogen test (BH2T) and the sulphasalazine/sulphapyridine method (SLZ) after the administration of a test meal of canned food mixed with sulphasalazine. Orocaecal transit time was defined as the time taken from the oral administration of the test meal to the time when the first portion of the meal reached the colon. In five of the dogs the OCTTs assessed by the BH2T were shorter than those measured by the SLZ method by 30, 15, 45, 30 and 45 minutes. However, the median OCTT assessed by the BH2T (135 minutes, range 120 to 195 minutes) was not significantly different from that measured by the SLZ (180 minutes, range 150 to 210 minutes) and was highly correlated with it (r = 0.94, P = 0.016). The sixth dog maintained baseline hydrogen and plasma sulphapyridine readings throughout the monitoring period and the OCTT could not be measured.

Palliative intracavitary carboplatin therapy in a cat with suspected pleural mesothelioma

A 12-year-old neutered male oriental shorthair cat was referred to the Animal Health Trust for investigation of pleural effusion. Ultrasonography revealed marked irregular thickening of the pleural surface of the cranial and caudal mediastinum. Cytological examination of the pleural fluid and fine needle aspirates of the thickened pleura suggested a diagnosis of mesothelioma. Following complete drainage of the thoracic cavity under ultrasound guidance, 180 mg/m2 carboplatin diluted in 60 ml sterile water was infused into the pleural space (30 ml in each hemithorax). This resulted in complete resolution of clinical signs for 34 days (having required thoracocentesis on four occasions in the preceding 4 weeks). The procedure was repeated using 200 mg/m2 carboplatin, and there was a further 20-day period where the cat was free of clinical signs. Further treatment was declined and the cat was euthanased 120 days after initial presentation. This is the first report of successful palliative chemotherapy for suspected feline mesothelioma and suggests that intracavitary carboplatin could be considered in tumours affecting the pleural cavity.

Polymerase chain reaction survey of feline haemoplasma infections in Greece

The aim of this study was to use real-time polymerase chain reaction assays to determine the prevalence of three haemoplasma species in cats from Greece and to evaluate possible associations between haemoplasma infection and age, gender, feline immunodeficiency virus/feline leukaemia virus (FIV/FeLV) status and packed cell volume (PCV). Ninety-seven cats (24 ill anaemic, 55 ill non-anaemic, 18 healthy non-anaemic) were included in the study. Twenty cats (20.6%) were haemoplasma positive; seven cats were infected only with Mycoplasma haemofelis, 10 were infected only with ‘Candidatus Mycoplasma haemominutum’ and three were co-infected with M haemofelis and ‘Candidatus M haemominutum’. ‘Candidatus Mycoplasma turicensis’ was not detected. Haemoplasma infection was associated with older age (P=0.019). M haemofelis infection tended to be more common in anaemic cats (P=0.058). No association between gender and haemoplasma infection, or haemoplasma relative copy number and PCV, was detected. Retroviral infection rates were very low with only one FeLV proviral positive cat found.

Measurement of thyroxine and cortisol in canine and feline blood samples using two immunoassay analysers

OBJECTIVES The AIA-360 (Tosoh Corporation) is an automated immunoassay analyser. The aims of this study were to estimate the precision of thyroxine and cortisol AIA-360 immunoassays in canine and feline samples and to compare the results produced with those obtained by a chemiluminescence analyser (Immulite® 1000, Siemens). METHODS Blood samples from 240 clinical cases (60 dogs and 60 cats for both thyroxine and cortisol) were analysed using both instruments. RESULTS Deming regression calculations showed excellent correlation (thyroxine, canine rs  = 0 · 94, feline rs  = 0 · 97; cortisol, canine rs  = 0 · 97, feline rs  = 0 · 97). Agreement between the two instruments was examined by Bland-Altman difference plots, which identified wide confidence intervals and outliers for thyroxine (canine n = 6, feline n = 4) and cortisol (canine n = 3, feline n = 4) results. Inter/intra-run precision of the AIA-360 was excellent for both cortisol and thyroxine (coefficients of variation <7%). CLINICAL SIGNIFICANCE The instrument showed excellent correlation for cortisol and thyroxine in canine and feline samples demonstrating that the AIA-360 can be used in clinical practice. The agreement studies suggest that the results from the AIA-360 cannot be used interchangeably with those generated by the Immulite 1000 and should be interpreted using reference intervals that have been established specific to the AIA-360.

Serum thymidine kinase activity in clinically healthy and diseased cats: a potential biomarker for lymphoma

The thymidine kinases are enzymes that convert deoxythymidine to deoxythymidine monophosphate and have a function in DNA synthesis. Rapidly proliferating cells will have higher levels of thymidine kinase. Serum thymidine kinase activity (sTK) is a useful tumour marker in humans and dogs, with utility as a prognostic indicator in lymphoma. In the current study serum samples were collected from 49 clinically healthy cats, 33 with lymphoma, 55 with inflammatory disease and 34 with non-haematopoietic neoplasia (NHPN). sTK was measured using a radioenzyme assay and a reference interval (1.96 × SD) was established from the clinically healthy cats (<5.5 U/l). Mean sTK activity for healthy cats was 2.2 U/l (range 0.8–8.4, ± SD 1.7). Mean sTK activity for cats with lymphoma was 17.5 U/l (range 1.0–100.0 SD ± 27.4). Mean sTK activity for cats with NHPN was 4.2 U/l (range 1.0–45.0, SD ± 8.6). Mean sTK activity for the inflammatory group was 3.4 U/l (range 1.0–19.6, SD 3.9). Cats with lymphoma had significantly higher sTK activity than healthy cats or cats with inflammatory disease (P <0.0001) and cats with NHPN (P <0.0002). sTK activity is a potentially useful biomarker for feline lymphoma and further study is required to assess its utility as a prognostic indicator.

Measurement of ionized calcium in canine blood samples collected in prefilled and self-filled heparinized syringes using the i-STAT point-of-care analyzer

BACKGROUND Heparinized syringes are commonly used with point-of-care analyzers (eg, i-STAT) to measure ionized calcium (iCa(2+)); however there is little information about the validity of their use in canine patients. OBJECTIVE To examine the suitability of prefilled (40 IU heparin/mL) and self-filled (150 IU heparin/mL) heparinized syringes for iCa(2+) measurements using the i-STAT analyzer. METHODS Forty-seven blood samples were collected from 41 canine patients. Two milliliters of blood were collected into a 2-mL nonanticoagulated (NA) syringe, a commercially available preheparinized (PH) syringe (dry calcium-balanced lithium heparin, 40 IU/mL), and a 2-mL self-filled heparinized (SH) syringe (liquid sodium heparin, 150 IU/mL). iCa(2+) was measured in the sample using the i-STAT analyzer and a wet-reagent analyzer (KoneLab 30i) used as the reference instrument. Data were analyzed using paired t-tests, Pearson correlation coefficient, and Bland-Altman difference plots. RESULTS There was no significant difference between the results obtained from NA and PH syringes using the i-STAT analyzer, and the correlation was excellent (r =.97). The i-STAT values from the SH syringes (mean, 1.07 mmol/L) were significantly lower (P<.001) than those from the NA syringes (mean, 1.38 mmol/L). iCa(2+) was significantly higher with the i-STAT analyzer than with the KoneLab analyzer for both the PH (mean i-STAT, 1.38 mmol/L vs mean KoneLab, 1.30 mmol/L) and SH (mean i-STAT, 1.07 mmol/L vs mean KoneLab, 1.03 mmol/L) samples. CONCLUSIONS Blood samples collected in the PH syringes used in this study can be used interchangeably with those collected in NA syringes for measuring iCa(2+) using the i-STAT analyzer. SH syringes with high-concentration heparin products (5000 IU/mL) are unsuitable for measuring iCa(2+) because they cause clinically significant underestimations. Although there was good correlation between the i-STAT and KoneLab analyzers, the results should be interpreted using analyzer-specific reference intervals.

The effect of short-term dietary fibre administration on oro-caecal transit time in dogs

SummaryThe effect on oro-caecal transit time of addition of soluble (guar gum) and insoluble (wheat bran) fibre to diets fed to seven healthy dogs at levels that have been shown to reduce post-prandial hyperglycaemia was assessed. Oro-caecal transit time was measured by sequential analysis of exhaled breath hydrogen levels. Median oro-caecal transit times were 105 min (range: 45 to 135), 113 min (range: 53 to 203) and 105 min (range: 75 to 195) after administration of a 770 g standard meal of canned food alone, or with the addition of 7.7 g wheat bran or guar gum, respectively. There was no significant difference between the oro-caecal transit times (p=0.964) for the different diets when the results for all the dogs were pooled. The difference between diets was statistically significant for both maximal post-prandial hyperglycaemia (p=0.035) and area of post-prandial hyperglycaemia under the post-prandial glucose curve (p=0.006).