CHAPTER III MATERIALS AND METHODS The present study entitled Studies on Categorisation of Prevalent Diseases and Clinical

Biomedical Profile of Geriatric Dogs was conducted by using the methodology discussed under the following heads: 3.1 Place of study The study was conducted at Small Animal Clinic and Department of Veterinary Medicine, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana. 3.2 Nature of Work The study was conducted on the clinical cases of dogs which were brought to the Out Patient Department of Small Animal Clinic. 3.3 Selection of Animal for Study Dogs of seven years of age or above, presented at Out Patient Department, Small Animal Clinic, were selected to be screened. Animals were subjected to comprehensive clinical examination. Special examinations were conducted on the basis of organ/system suspected to be involved as per requirement of the case. Subsequently the cases were categorised into various disease conditions (organ wise: kidney, liver conditions, heart problems, skin, ocular, GIT conditions, respiratory problems, musculoskeletal conditions in geriatric dogs on basis of age, Breed and Sex of the animal.

3.4

CLINICAL EXAMINATION

3.4.1 Client information Case number, date of presentation, name, address and phone number of the dog owner were recorded in Geriatric Case Record 3.4.2 Signalment Patient specific data i.e. case number, age, breed, sex; weight was recorded in Geriatric Case Records 3.4.3 History History was recorded with respect to appetite, exercise intolerance, lethargy, abdominal discomfort, vomiting, diarrhea, fever, weakness, previous sign of illness, dietary history, any history of trauma ,vaccination, deworming status, history of weight gain or weight loss, polydipsia, polyuria, polyphagia, history of pica etc. 3.4.4 Detailed physical examination A complete physical examination of the animal was carried out as proposed by Nelson and Couto (2009).Various parameters like rectal temperature, respiration rate, heart rate, pulse rate, body condition and colour of the visible mucus membrane were recorded. A detailed percussion and palpation of abdomen was performed for presence of any pain, fluid or abnormal mass in the abdomen. A thorough auscultation of the thorax for the nature of cardiac rhythm, rate, and adventitious sound was also performed and recorded. In dogs with abnormal cardiac auscultation findings, ECG was performed and if possible Echocardiography was done. Based on the above observations, a tentative diagnosis was drawn.

3.5 BLOOD EXAMINATION 3.5.1 Blood collection The animal was properly restrained on the lateral recumbency. The blood was aseptically collected from the cephalic vein or the recurrent tarsal vein. The vials with disodium EDTA as anticoagulant were used for preservation of blood for hematology and blood protozoa, Sodium fluoride vials were used for Glucose estimation. Blood samples collected in sterile syringes were allowed to clot for 2 hours and were centrifuged at 2500 rpm for 3 minutes for separation of serum. Serum samples were collected and preserved at -200 C for estimation of biochemical parameters. 3.5.2 Haematological analysis The following parameters were estimated was estimated using fully automated haematology analyser.1 a) Hemoglobin (g/dL) b) Total leucocyte count (TLC) c) Total erythrocyte count (TEC) d) Packed cell volume (PCV) e) Platelet count Differential leukocyte count (DLC) was done using Leishmans stain as per method suggested by Jain (1986). 3.5.3 Biochemical analysis
Serum samples were analysed according to the system involved for determining

activities of, alanine aminotransferase (ALT), aspartate aminotransfrerase (AST), alkaline phospahatase (ALKP), gamma glutamyl transferase (GGT) and concentrations
1

ADVIA 2120 Hematology system, Siemens Healthcare Diagnostics Inc., USA

of Total Bilirubin, blood urea nitrogen (BUN), creatinine, total plasma protein and albumin., Calcium, Phosphorus, Glucose, Cholesterol, Sodium and potassium. These analyses were performed using fully automated chemistry analyser2 with 3.54 Blood/fecal examination for parasites Blood and fecal samples were examined for the presence of any parasitc infestatation as per method of Soulsby (1982). Blood was collected in EDTA containing vials and the samples were evaluated for haemoprotozoan infections reagent kits.3 3.6 URINE ANALYSIS Urine of geriatric dogs (when required) was collected in plastic vials by catheterization or cystocentesis aseptically. The urine was subjected to specific gravity and chemical analysis wherever required. Specific gravity was examined using dipsticks1 and presence of proteins, glucose, bilirubin, ketones, blood, urobilinogen, nitrite, leucocytes was detected using dipstick method4,5 3.7 ELECTROCARDIOGRAPHY Echocardiography was performed in animals suspected of cardiac abnormality on the basis of physical examination and ausculatation findings (rate, rhythm, adventitious sounds). Dogs were restrained in right lateral recumbency on wooden table. ECG was recorded by using Baileys hexaxial lead system. Limb leads were applied on elbow joint on both forelegs and slightly above stifle joint on the hind leg. Recordings were performed at paper speed of 50 mm/sec and amplitude of 10 mV. All the abnormalities were detected from lead II tracing.

2 3

Johnson & Johnson VITRS 750Xrc & Fully automated chemistry system. Johnson & Johnson diagnostic kits, Mumbai, India 4 Multistix 10 SG Reagent Strips for Urinalysis, Siemens Healthcare Diagnostics Inc., USA 5 CLINITEK STATUS analyzer, Bayer Healthcare LLC

3.8

RADIOGRAPHY A lateral and ventrodorsal radiograph of chest was taken to visualize the size, shape of

the heart and the cardiac silhouette in dogs suspected for cardiac disease. The lung fields were evaluated for any abnormality or metastasis. Lateral radiograph of abdomen were taken to detect any abnormal mass, uroliths, pyometra, nephroliths, hepatomegaly, spleenomegaly, prostate enlargement, spondylosis etc. 3.9 ULTRASONOGRAPHY Dogs required were subjected to abdominal ultrasonography for visualisation of different organs like liver, gall bladder, spleen kidney, urinary bladder, prostrate and uterus. Organs were examined using a concept/MCV Veterinary Ultrasound Scanner (a gray scale, real time B mode scanner). A 3.5 MHz microconvex or 7.5 MHz microconvex linear array transducer was used depending on the patient. 4.0 HISTOPATHOLOGICAL EXAMINATION

Liver biopsy was collected by ultrasound guided trucut biopsy needle in four dogs and the samples were procured in 10% formalin solution. Histopathological evaluation was done using H&E stain. 4.1 POSTMORTEM EXAMINATION Postmortem examination was conducted on 3 dogs which died during treatment or were brought dead. Tissue samples were collected in 10% formalin and processed for further histopathological examination for reaching to a confirmatory diagnosis.

DIAGNOSIS A clinical diagnosis was formed on basis of clinical signs, complete blood count and various biochemistry parameters of the blood samples in suspected cases. Confirmed diagnosis where possible was obtained with, Radiography, Echocardiography and biopsy or postmortem examination. Hundred geriatric dogs were examined and they were categorised into twelve major groups based on the etiology viz. urinary diseases (n=46), Tumors (n=20), hepatic disorders (n=14), musculoskeletal conditions (n=15), heart (n=8), reproductive problems (n=9), respiratory problems (n=5), skin conditions (n=5), ocular conditions (n=5), ear affections (n=4) and endocrine abnormalities (n=5), peritonitis(n=3) Urinary system disorders were further divided into Viz. Renal failure (n=36), urolithiasis (n=6), and Cystitis (n=4) which were diagnosed on the basis of hematobiochemical analysis, urine analysis, radiography and ultrasound techniques. Clinical diagnosis of tumors were made on basis of Ultrasound and confirmatory diagnosis were made on basis of histopathology by taking swabs, aspirations from the swelling, biopsy and postmortem examination analysis. Fifteen cases of musculoskeletal disorders were further divided into Viz. spondylosis (n=10), degenerative joint disease (n=2), hip dysplasia (n=1) and myositis (n=1).The cases were confirmed on basis of radiographic images. Reproductive cases were further diagnosed into pyometra (n=3) and prostrate affections (n=6) diagnosed on basis of radiographic images and confirmed by ultrasonography. Clinical diagnoses of Endocrine problems were diagnosed on basis of biochemical parameters.

Clinical diagnosis of respiratory system problems were diagnosed on the basis of clinical signs and radiographic images. Effect of age on skin, eye and ear affections conditions was diagnosed by clinical signs. Cardiac disorders were diagnosed by clinical signs, radiography, ECG and echocardiography. On the basis of these techniques cardiac disorders were diagnosed as DCM, pleural effusions and ventricular hypertrophy. Out of one hundred and forty dogs 14 dogs were diagnosed with hepatopathies and Among 14 dogs, 6 dogs each were diagnosed of chronic hepatitis and hepatosis each, one dog each were diagnosed cholecystitis and cirhossis each. These were diagnosed on the basis of clinical signs, hematology, enzymology, liver biopsy and histopathology where ever possible.