Biochemistry

Vital organs such as the heart, liver, kidneys, and lungs communicate with each other through chemical regulators synthesised in the nucleus or in the cytoplasm of the cells in these organs. Analyses of these biochemical substances are crucial in assessing the cellular pathophysiological status of the organs in gastrointestinal, genitourinary, cardiovascular, respiratory, reproductive, and musculoskeletal systems and hence the health of these organs are assessed. Our laboratory offers common clinical chemistry test panels for horses. This includes, comprehensive profile: Glucose, Blood Urea Nitrogen (BUN), Creatinine, Total Protein, Albumin, Globulins, Phosphorus, Sodium, Chloride, Potassium, Total CO2, Anion gap, Total and Ionized Calcium, Total Bilirubin, Alkaline Phosphate, ALT, and GGT. Others are renal, liver and muscle profiles, inflammation assessment (Total protein, Albumin, Globulin, Amyloid A in serum), electrolytes balance assessment (Na, Cl, Mg, PO4, Ca, K) and equine weight loss profile (AP, GGT, Bile Acid, Albumin, TP, Total globulin, A/G ratio and CBC). Equine blood gas panel includes testing pH, pCO2, pO2, tHb, K+, Na+, cCa2+, Cl-, Glu, Lac, Crea, Urea, tBil in the whole blood. Cholesterol, Triglycerides HDL and LDL are also available on demand. Urinalysis is carried out using urine dipstick test, total protein and specific gravity are performed by refractometer along with direct microscopy, stained and unstained urine cytology and urine culture.

Blood and Urine Protein Electrophoresis

Blood and urine are composed of various protein moieties integral indicators in both health and disease conditions. Separating and quantifying these proteins from a serum and urine sample aid in diagnosis of infectious, allergic, nutritional and parasitic diseases in horses. Serum protein electrophoresis is also a valuable tool for the diagnosis of combined immunodeficiency in Arabian horses. Total Protein, Pre-Albumin, Albumin, globulins (Alpha globulin, Beta globulin, Gamma globulin) are assessed in automated capillary electrophoresis machines.

Parasitology

This section is simple yet vital in maintaining the well-being of horses by detecting external and internal parasites or parasitic ova in the fecal or skin scraping samples. Internal parasites being one of the common health concerns among horses wherein intestinal parasite like ascarids, large and small strongyles and tapeworms may cause weight loss, anemia, diarrhea and colic. Direct mount microscopy, fecal flotation, fecal sedimentation for lung worms, fecal sand test, Knott's test for equine microfilaria, fecal worm egg reduction test, Baermann's test for nematode larvae and quantitation of egg parasites in the stool using McMaster fecal egg count are the routine tests at the laboratory. Peripheral blood smears for suspected equine piroplasmosis and cryptosporidiosis by are examined using staining and microscopy. Skin scraping KOH test for checking external parasites and dermatophytes is available. Fecal occult blood test and nematode larvae culture test are in line to be added to the laboratory portfolio.

Molecular Diagnostics

Our molecular diagnostic laboratory is equipped with the state-of-the-art equipment and facilities, including automated extraction and qPCR. Wide ranges of viral, bacterial, fungal, and parasitic infections are identified in our PCR laboratory by using qPCR, which is a powerful technique and used for rapid detection of pathogens, including those, which are difficult to culture. Advantages of Real-Time PCR include greater sensitivity, reproducibility, rapid analysis, better control of quality in the process, and a lower risk of contamination.

Microbiology

The microbiology laboratory performs phenotype profiling for wide range of bacteria and fungi of veterinary importance using standard microbiological techniques of culturing, incubating, isolation and identification with automated biochemical and proteomic approaches (Matrix Assisted Laser Desorption Ionization Time of Flight Mass Spectrometry). The laboratory is equipped with automated microbial detection system with quick and efficient microbial detection in blood or other body fluids (CSF, plural, pericardial and joint fluids) using aerobic and anaerobic culture media bottles.

Isolation, ID and AST of oxygen sensitive anaerobic bacteria are performed in atmosphere controlled anaerobic chamber. Different enrichment culture procedures are available for the identification of organisms. Both automated minimum inhibitory concentration (MIC) and manual disc diffusion method based on zone of bacterial inhibition around the antibiotic disc, are available for antimicrobial susceptibility under CLSI and EUCAST guidelines. The laboratory receives a wide range of clinical and environmental samples for bacterial and fungal isolation, identification and antimicrobial susceptibility testing (AST) on a daily basis. Various test plans are available for diagnosis and research.

Endocrinology

The endocrine glands: pituitary, thyroid, parathyroid, pancreas, ovaries and testes produce hormones necessary for balanced functions in metabolisms, general growth, reproduction, and the body’s response to combat stress. Our diagnostic tests for the endocrine system are building up. We use automated fluorescence enzyme immunoassay for adrenocorticotropic hormones, progesterone, testosterone, free thyroxine, insulin and cortisol. Adiponectin, Anti-Müllerian hormone, leptin, estradiol, and inhibin using equine specific enzyme-linked immunosorbent assay are in line to be added to the endocrinology lab.

Histopathology

In histopathology, physical alterations to the cellular architecture in response to infectious, necrotic or neoplastic insult to the tissue samples are observed under the microscope. Currently, common histopathology services are available for examining endometrial biopsies, visceral organ tissues samples from post-mortem cases, and samples from the tumors. Apart from routine H&E stain,various other stains are also used on the processed fine sections of the tissues for examination. The histopathology laboratory is equipped with Immuno-histo chemistry and in situ hybridization staining equipment for diagnosis and research.

Cytopathology

In comparison to histopathology techniques, diagnostic cytology provides a quick and relatively non-invasive way to evaluate the cellular reaction to infection, inflammation and neoplasms. Important factors affecting the cytological picture are the type of injury or the type of invading pathogen, the site of infection process, its route of entry and the stage of the disease i.e. acute and chronic recurrent phase. The types of cells such as epithelial cells, neutrophils, lymphocytes, monocytes, bacteria or fungi are evaluated. The role of specific cells such as in ensuing immune response, phagocytic activity of monocytes and their activated forms in removing cell debris and pathogenic organisms, the extent of tissue damage, the stage of disease and the prognosis are assessed. Currently, Diff Quick or Giemsa stain, Gram's stain, Papanicolaou and PAS stains are routinely used for diagnostic cytology.

Necropsy

A well-established necropsy service is available at EVMC carried out by well renowned pathologist with years of experience. Apart from regular necropsy for neonatal and adult horses, we also examine aborted fetuses and placentas to understand the plausible cause of death. Strict biosecurity measures are in place and efficient carcass disposal by a third party contractor is available. Presenting approved Necropsy Request Form is mandatory for carrying out the necropsy examination.

Regenerative Medicine Therapy

At the moment the laboratory offers platelet rich plasma (PRP) and Interleukin-1 Receptor Antagonist Protein (IRAP) are concentrated from the blood samples of in-house horse patients at EVMC. These therapies are getting popular for the treatment of musculoskeletal injuries and osteoarthritis in horses. Platelets are blood cells smaller than RBC; they play an important role in hemostasis and are considered to be the natural sources of growth factors. The growth factors are believed to be actively involved in wound healing and other regenerative processes in the wake of injuries. These growth factors are rich in concentrated platelet preparations. After preparation, these products are injected back into the same horses sampled to promote tissues repair in joint and soft tissue injuries. At the site of trauma and joint swelling inflammatory proteins such as cytokines and Interleukin-1 (IL-1) are released, IRAP therapy prevents IL-1 binding to the joint tissues minimizing the damaging effect of IL-1 in the affected joint. Other products such as stem cell therapy are in line to be added to EVMC laboratory. Regenerative medicine laboratory is following strict good manufacturing practices (GMP).