Axiom Veterinary Laboratory News Letter AUTUMN 2000

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Autumn 2000

"NEWSFLASH
New Axiom Multiuser Sample Box"
Topscreen for Cats
How to get your samples to us quickly and safely
Comings and Goings
What is in your Diagnosis?
Canine Hypothyroidism: Your questions answered
Investigation of suspected Bleeding Disorder
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Topscreen for Cats 

We are pleased to offer the only in-vitro allergy screening test for cats, TOPSCREEN CAT is available on its own (£26) or combined with flea saliva (£35.50). This is the same format that is available for the dog. Please make sure on your submission form that you clearly specify either a canine or feline Topscreen as different monoclonal antibodies are used for these two tests. The expanded ALLERCEPT allergy testing service using the Fc epsilon receptor technique continues to be available for both dogs and cats and immunotherapy preparations (IMMUCEPT) can also be prepared on prescription for both species. This makes Axiom Veterinary Laboratories the leading diagnostic laboratory for allergic skin disease in the UK.

To complement this expertise we are now offering an expanded endocrine service under the guidance of Dr Richard Dixon. Our range of endocrine assays now includes canine TSH and FT4 by equilibrium dialysis. This facility, if used in conjunction with our comprehensive cytology and microbiology services, should allow practitioners to investigate even the most challenging dermatological and endocrine cases. 

How to get your samples to us quickly and safely 
We are proud to announce that we have been re-certified under ISO 9002 for a further 3-year period. Possession of this mark means that we are rigorously inspected every six months with a longer inspection once every 3 years. It is this renewal inspection that we have passed with flying colours. This is a tribute to all members of staff and the high quality of service that we look to achieve.

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Comings and Goings  

As part of our effort to get results to you in a timely fashion we have been working with Verifac computers and their Congress information system so that we can send our results back through this system. If you have a Verifac computer system within your practice this means results can go straight back into patient files. If you do not possess a Verifac system but are running on a windows based system then the same software can be used to cut and paste results straight into patient files. If you are interested in this Technology please do not hesitate to contact us in the first instance.

As you know, our automated fax system has caused us some grief over the past 12 months. It is certainly working better than before but still not to our satisfaction. Unfortunately in an effort to make this piece of software work correctly some damage has been done to the typefaces and the access number has been moved from its original position. The software people are working on this and I apologise for the temporary poor quality of these reports. One annoying problem is that the phone number 778844 appears as 778811. Unfortunately this goes through to a very kindly old lady who knows nothing about clinical pathology! As she is a kindly lady she understands why this is happening but obviously we need to stop this nuisance. Please note the phone number is 01626 778844; it is an ISDN line, which means there are between 6 and 10 active incoming phone lines on this single number.

You will also note that all members of staff have a direct call number and this is listed both on our CD and the printed copies of our price list and so if you wish to speak to someone directly it is probably easier to use the direct line number.

Our printed fax line number of 01626 779570 is always very busy but there are other numbers you can use if you can't get through on this line and they are 01626 771447 and 01626 771448.

We can also send results back to you by Email; this is quick and efficient and bypasses the wretched fax software so the presentation is better as well!

All those who have received our new CD will I am sure see a tremendous improvement on last year's effort.
We are already working on the 3rd edition for 2002. This CD will load on the hard drive of an individual PC or indeed a server. It should produce an icon in windows and this icon can be minimised on the tool bar so the programme can remain running in the background during consultations. Not only does this CD-ROM contain our price list but also the various protocols that we have produced, particularly those relating to endocrine evaluation. There are some very basic haematological images included on the CD which you can print, which are intended to be helpful to your veterinary staff who are doing haematology. In addition these images are also available on a A4 poster to pin up on the wall of the practice lab. If you would like a poster and have not received one please contact the office.

If by chance your practice cannot operate a CD-ROM (there are some!), or does not have a windows operating computer system, we can provide you with a printed copy of both the price list and the guide. The price list is also available in HTML; Microsoft Word and rich text formats, the latter being particularly useful if you are working on a Unix based system. This information can be supplied to you on a floppy disc or as an email attachment. Please contact the office in the first instance with your requirements.

Martin Wheeler our Director of Graphics has been very busy designing our website, the launch of this web site is imminent and can be found at www.axiomvetlab.co.uk or axiomvetlab.com. As this is a public site certain veterinary information would be inappropriate and is not displayed. Over the next few months we will be "playing around" with certain ideas so the site is likely to continually change.

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What is in your Diagnosis?

A 3 year old male Hungarian visla presented with a 4 week history of polydipsia, polyuria, and progressive weight loss. Abdominal radiography confirmed the abdominal distension is due to ascites; no other radiographic abnormalities were detected.

A polydipsia profile reveals the following haematological and biochemical abnormalities:

Haematology: Erythrocytosis (RBC 10.37 x1012/l; PCV 0.69 l/l; Hb 24.0 g/dl),and mild neutrophilia (11.7 x109/l).

Biochemistry: Total plasma proteins 38.5 g/l; hypoalbuminaemia (9.9 g/l); hypocalcaemia (2.26 mmol/l); hypercholesterolaemia (11.7 mmol/l).

Urinalysis (catheterized sample): Mild haematuria, proteinuria (3+ on dipstick) and hyposthenuria (urine SG 1.005). Urine microscopy was otherwise unremarkable.

Peritoneal fluid analysis: The gross specimen appeared clear and colourless. Protein 1.8 g/l; SG 1.008; nucleated cells 0 x109/l; RBC 0 x1012/l. Only a few mesothelial cells were seen on the cytospin preparation.

On the basis of these results what would be your tentative diagnosis? What additional test(s) may be helpful? Please see page X for an interpretive summary of the laboratory abnormalities described above.

Answer to the case challenge
The hypocellularity, low protein content and SG are typical of a true transudate, hence the water-like appearance of this effusion. Hypoalbuminaemia can be associated with chronic liver disease, glomerulonephropathy and protein-losing enteropathy (PLE). With PLE globulins may also be significantly decreased which is not the case here. 3+ protein on a urine dipstick is likely to be significant given the low specific gravity and absence of an active urinary sediment. The fact that cholesterol s also increased provides additional support for a protein-losing nephropathy.

The extent of the urinary protein loss can be more precisely quantified by measuring the urine protein:creatinine ratio (since it has been shown that the UP:UC ratio on a single urine sample correlates well with measured 24 hour protein excretion). The UP:UC ratio in this case was 11.8 which is high (normal usually less than 1.0). Values of this magnitude are indicative of glomerular disease. Proteinuria, hypoalbuminaemia, hypercholesterolaemia and ascites or subcutaneous oedema are classical features of the nephrotic syndrome.

The erythrocytosis may simply be due to severe dehydration. Equally, it is possible that this dog had absolute polycythaemia secondary to renal disease. Secondary polycythaemia most frequently is associated with renal disease eg renal lymphoma, pyelonephritis etc.

Two images of reactive mesothelial cells in peritoneal fluid.

SOME BACKGROUND INFORMATION
The main glomerular diseases in the dog are immune-complex glomerulonephritis and amyloidosis. True autoimmune glomerulonephritis involving antibodies against glomerular basement membrane antigens is thought not to occur in the dog. Deposition of immune complexes in the capillary walls of the glomerulus ultimately leads to malfunction of the entire nephron. Frequently, however, during the earlier stages of the disease, nephron function remains adequate hence urea and creatinine may be within reference limits (as was the case here).

Glomerulonephritis can also be a manifestation of systemic lupus erythematosus (SLE). Generally, however, dogs with SLE show involvment of other body systems in addition to the kidneys eg joints (immune-mediated polyarthritis), skin and blood (immune-mediated anaemia and/or thrombocytopenia). Glomerulonephropathy may also be associated with leishmaniasis and, less frequently, ehrlichiosis.

Amyloidosis may occur as a primary systemic disorder in Shar peis. Secondary reactive amyloidosis, involving the deposition of amyloid A (AA) protein, is associated with chronic inflammatory disease and neoplasia. Differentiation of renal amyloidosis from immune-mediated glomerulonephritis requires histological examination of a renal biopsy.

Dogs with glomerulonephritis usually do not develop oedema or ascites until the albumin concentration is well below 15 g/l. Both albumin and globulins may be lost in urine as the disease progresses (urine protein electrophoresis may be helpful in this respect). Hypercoagulability develops as a result of increased platelet adhesion and aggregation, and the urinary loss of antithrombin. Pulmonary thromboembolism is therefore an important potential complication of glomerulonephritis in dogs. Systemic hypertension occurs in over 80% of dogs with glomerular disease and may lead to retinal haemorrhage, detachment and papilloedema, and approximately 60% have hypercholesterolaemia which may also contribute to platelet hyperaggregability.

Treatment: The prognosis for dogs with suspected immune-mediated glomerulonephritis is extremely guarded; any response tends at best to be short lived. Azathioprine, chlorambucil, cyclophosphamide and cyclosporine have been used either experimentally or in clinical situations with variable and often disappointing results. The administration of corticosteroids is controversial since it has been suggested that they may actually increase the proteinuria and promote azotaemia. Aspirin may be beneficial both for its anti-inflammatory and antithrombotic properties if given at low doses (0.5 mg/kg once or twice daily). A low sodium protein restricted diet is advised to minimize the risk of sodium retention and systemic hypertension. The use of ACE inhibitors eg enalapril has yet to be fully investigated.

Aknowledgments: Axiom Veterinary :Laboratories would like to thank Helen Harper of the Shepton Vet Group, Shepton Mallet, Somerset for allowing us to publish details of this interesting case.

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Canine Hypothyroidism: Your questions answered

The reliable confirmation of hypothyroidism in dogs continues to pose one of the greatest diagnostic challenges in practice. We now offer a thorough range of thyroid tests including free T4 by equilibrium dialysis (fT4d) and thyroglobulin antibodies (TgAb) to help with these cases. However, being an area of some confusion, we thought it would be useful to answer the five most common questions relating to testing for canine hypothyroidism.

1) "What is the best profile to test for hypothyroidism ?"
We offer a standard profile (total T4 and cTSH), a premium profile (total T4, fT4d and cTSH), and a comprehensive profile (total T4, fT4d, cTSH and TgAb). Since there is no single perfect test for hypothyroidism we recommend our comprehensive profile. This is now very widely used and is designed to maximize your chances of making an accurate diagnosis. However if cost is a major consideration then both the premium and standard profiles offer excellent value for money.

2) "What are thyroglobulin antibodies and when should I test for them ?"
Dogs with lymphocytic thyroiditis (the most common cause of hypothyroidism) produce antibodies to normal thyroid tissue, including thyroglobulin. The identification of TgAb, using the latest method, is a highly specific marker for thyroid disease.

3) "Isn't measurement of total T4 alone good enough ?"
Total T4 is depressed by virtually all non-thyroidal illnesses, drug therapy and is even subnormal in healthy dogs at various times during the day. Consequently finding a decreased total T4 concentration does not confirm hypothyroidism. Instead of requesting total T4 measurement alone, we recommend using one of our thyroid function profiles, which are designed to help differentiate genuine hypothyroidism from these other non-thyroidal influences.

4) "Should I try trial thyroid replacement therapy ?"
We only recommend this approach as a last resort when all other options have been exhausted. After trial therapy has been attempted, it is very difficult to assess thyroid status at a later date. These cases will often return to haunt you at a later date!

5) "How should I monitor a dog receiving thyroid replacement therapy?"
Blood should be collected for total T4 and cTSH estimation to monitor treatment. It is important that the sample is collected six hours after the medication has been given. A 6 hour post-pill TT4 value less than 35 nmol/l for a dog receiving once daily medication indicates thyroid replacement is inadequate. TSH is generally at the low end of the limit of detection of the assay.

Investigation of suspected Bleeding Disorder

We now have a supply of Simplate II devices for measuring buccal mucosal bleeding time (BMBT) and blood tubes for measuring activated clotting time (ACT). These can be ordered through our diagnostic support team. Both the BMBT and ACT should be performed in conjunction with our haemorrhage screen which includes a complete blood count, one stage prothrombin time (OSPT) and activated partial thromboplastin time (APTT).

Activated clotting time
This test uses 2ml of whole blood which is added to a special tube containing diatomaceous earth. It evaluates the intrinsic and common pathways (Factors XII, XI, IX, VIII, X, V and II) of the coagulation cascade. ACT will be prolonged if there is more than a 70%-75% decrease in any one of these Factors. A clot should form in 60-100 seconds (dog) or less than 60 seconds (cat) if the tube is incubated at 37oC. Note that after 60 seconds the tube should inverted every 5 seconds until a clot forms. Potential causes of prolonged ACT include warfarin toxicity, haemophilia and disseminated intravascular coagulation.

Buccal mucosal bleeding time
Buccal mucosal bleeding time (BMBT) can be measured using the Simplate II device. This makes two linear incisions in the buccal mucosa and the time taken for complete cessation of bleeding is determined. Filter paper is used to absorb excess blood every 30 seconds but care should be taken not to touch the incision. Bleeding time in normal dogs and cats is 1.7-4.2 minutes (mean 2.6 min). BMBT will be prolonged with thrombocytopenia and platelet dysfunction (eg associated with aspirin therapy or von Willebrand's disease).

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Axiom Veterinary Laboratories Ltd.
The Manor House, Brunel Road, Newton Abbot, Devon TQ12 4PB, UK.
TEL: +44 (0)1626 355655 FAX: +44 (0)1626 357750/1 E-mail:admin@axiomvetlab.co.uk
 

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