Diagnostic parasitology

Diagnosis of parasitic infections in small animals is challenging, interesting, and fun, and involves the recognition of parasite stages based on size, morphology, color, and movement. Size and morphology are the major diagnostic parameters, and a calibrated microscope is the essential tool in the diagnostic laboratory. The fecal flotation technique is used routinely for the diagnosis of most parasites that reside within the gastrointestinal tract. The specific gravity and type of solution used often will influence the results obtained. A sugar flotation solution with a specific gravity of 1.27 is one that will float a majority of parasite stages with minimal distortion. Other techniques for diagnosis of specific gastrointestinal parasites include the direct smear and the merthiolate-iodine-formalin preservative method for diagnosis of Giardia sp, the Baermann technique for diagnosis of lungworm and other live larvae, the direct sedimentation technique for diagnosis of trematode eggs, the ether-formalin-sedimentation technique for diagnosis of trematode eggs and concentrating protozoan cysts from feces with high fat content, and the McMaster technique, a dilution modification of the fecal flotation technique. Techniques for evaluation of parasites in blood include the Knott's test, hematocrit method, and direct blood smear for diagnosis of microfilariae and thick and thin stained blood smears for diagnosis of Babesia sp, Haemobartonella sp, Cytauxzoon felis, Hepatozoon canis, and Trypanosoma cruzi. Tissue impression smears are used commonly for the diagnosis of Leishmania sp in dogs and biopsy specimens or aspirates are used for the diagnosis of Pneumocystis carinii. Diagnosis of acute toxoplasmosis can be accomplished with peritoneal and thoracic fluids, and organisms occasionally are detected in blood or spinal fluid. Serological tests for many parasitic diseases have been developed and often are used as supportive diagnostic tests.     

Treatments and outcomes of severe hemorrhagic enteritis in three eastern bongos (Tragelpahus eurycerus isaacii) at the Singapore Zoo

Three eastern bongos (Tragelaphus eurycerus isaaci) presented acutely with hemorrhagic diarrhea at the Singapore Zoo, thought to be caused by a mouldy batch of hay. Repeated fecal tests were negative of parasites and common gastrointestinal bacteria including salmonella and campylobacter. The diarrhea resolved for all individuals after a week of leaf-only diet. However, 2 individuals developed signs of colic. Both animals were anesthetized for examination including blood tests and imaging studies. The findings were consistent of gastrointestinal ileus and a possible impaction. With intensive treatment involving repeated sedations for fluid therapy administration and treatments for gastrointestinal impaction, one individual eventually made a full recovery, but the other individual died due to septic peritonitis secondary to a rupture in the spiral colon. Persistent supportive therapy may be vital in treating severe gastrointestinal disease in this species.     

Intestinal hyperammonaemia in horses

Intestinal hyperammonaemia in horses is a syndrome putatively caused by increased intestinal production and/or absorption of ammonia that clinically manifests as severe neurological abnormalities often in conjunction with signs of gastrointestinal dysfunction but without evidence of hepatic compromise. Ingestion of Black Locust (Robinia pseudoacacia) is a novel primary condition that may predispose to development of the syndrome as described in this issue. Treatment of the condition is symptomatic and aimed at decreasing blood ammonium concentrations by reducing further production and absorption, reducing brain oedema and preventing self‐inflicted trauma. Prognosis for survival is guarded, particularly in cases with severe gastrointestinal lesions, and is influenced by the ability to control clinical signs.     

Triage and approach to the acute abdomen

The presentation of a patient with acute abdominal pain often is both a diagnostic and therapeutic challenge for the veterinary practitioner. Rapid physical examination and assessment of the patient's cardiovascular status are critical to initiating appropriate and often aggressive medical management as diagnostic tests are being performed. A number of diagnostic tests can be performed, including complete blood count, serum biochemistry profile, serum amylase and lipase, abdominal radiographs, abdominal ultrasound, abdominal paracentesis or diagnostic peritoneal lavage, cytology, and fluid analysis. In some cases, surgical exploration may be indicated, depending on each patient's clinical signs and response to therapy. Of the most important aspects of medical management of the patient with acute abdominal pain is to maintain oxygen delivery to tissues and to treat aggressively for pain or discomfort. The initial triage and approach to the patient with acute abdomen are discussed.     

Pharmacologic management of feline hyperthyroidism.

Radioiodine is considered the treatment of choice for hyperthyroidism, but in some situations, methimazole therapy is preferred, such as in cats with preexisting renal insufficiency. Unfavorable outcomes from methimazole are usually attributable to side effects, such as gastrointestinal upset, facial excoriation, thrombocytopenia, neutropenia, or liver enzyme elevations. Because restoration of euthyroidism can lead to a drop in glomerular filtration rate, all cats treated with methimazole should be monitored with blood urea nitrogen and creatinine levels in addition to serum thyroxine (T(4)) and a complete blood cell count. Transdermal methimazole is associated with fewer gastrointestinal side effects and can be used in cats with simple vomiting or inappetence from oral methimazole. Hypertension may not resolve immediately when serum T(4) is normalized, and moderate to severe hypertension should be treated concurrently with atenolol, amlodipine, or an angiotensin-converting enzyme inhibitor.     

Abdominal irradiation as a rescue therapy for feline gastrointestinal lymphoma: a retrospective study of 11 cats (2001-2008)

In this retrospective study medical records of 11 cats with gastrointestinal lymphoma were evaluated to determine the efficacy of radiation therapy when used in a rescue therapy setting. All cats had relapsed or resistant lymphoma. Two fractions of radiation were delivered over 2 days for a total of 800cGy. Acute effects of radiation were not noted, except one cat that had a self-limiting loss of appetite. Response was noted in 10/11 cats. Median survival post-radiation therapy was 214 days and the overall median survival in this study was 355 days. This study suggests that abdominal irradiation for feline gastrointestinal lymphoma was well tolerated and may contribute to a positive clinical response.     

Nonsteroidal anti-inflammatory drugs induce hypermotilinemia and disturbance of interdigestive migrating contractions in instrumented dogs

Nonsteroidal anti-inflammatory drugs (NSAIDs) induce gastric ulcers due to inhibition of prostaglandin production. Prostaglandins have an influence on physiological gastrointestinal motility, but the relationships between NSAID-induced gastric ulcer, gastrointestinal motility and motilin are unknown. Fifteen dogs were allocated randomly to three groups in which either gelatin, meloxicam or indomethacin was administered. Fecal occult blood and gastrointestinal motility were monitored continuously for 6 days. In addition, analyses of the plasma motilin concentration, gastrointestinal endoscopy and gastric emptying, and detection of motilin cells were performed. Gastrointestinal motility was disturbed in the indomethacin group, presenting as disappearance of interdigestive migrating contractions (IMCs) 3 days before gastric ulcers were detected. Delayed gastric emptying and hypermotilinemia were observed significantly more often in the indomethacin group compared with the other groups. Motilin cell-crypt/villi ratio in the indomethacin group significantly decreased in the duodenum and jejunum, compared with the other groups. No significant changes in any tests were observed in the meloxicam group, when compared with the gelatin group. These findings suggest that the disturbance of IMCs caused by hypermotilinemia, with changes in motilin cell distribution, and delayed gastric emptying induced by indomethacin may contribute to the development of gastric ulcers.     

Laboratory procedures for the diagnosis of gastrointestinal tract diseases of dogs and cats

An increasing number of laboratory tests are available for diagnosis of gastrointestinal tract diseases in dogs and cats. Use of these tests can lead to more accurate and rapid diagnoses. This review discusses laboratory tests, both new and old, and the role they currently play in the evaluation of animals presented with gastrointestinal problems. A minimum database helps assess the severity of the disorder, detect extra-gastrointestinal causes of problems and assists in formulating diagnostic and therapeutic plans. Faecal examination remains one of the most important diagnostic procedures in the investigation of gastrointestinal problems. Zinc sulphate faecal flotation is an excellent routine screening technique for helminth and protozoal infections, including giardiasis. Rectal cytology can assist in the diagnosis of large bowel disorders. Interpretation of faecal immunodiagnostic tests is hampered by insufficient knowledge of test sensitivities and specificities. Routine faecal cultures are not warranted and faecal occult blood tests are rarely indicated. Serum tests for gastric inflammation are now under development. The serum trypsin-like immunoreactivity test remains the gold standard technique for the diagnosis of exocrine pancreatic insufficiency. Breath hydrogen tests can be helpful in assessing the functional relevance of mild abnormalities in small-bowel biopsy specimens. Subnormal concentrations of serum cobalamin appear to be more specific indicators of gastrointestinal disease in cats than in dogs. Tests for small intestinal bacterial overgrowth remain controversial and assessment of gastrointestinal permeability has yet to prove its value in the diagnostic assessment of companion animals with gastrointestinal problems. Faecal alpha1-protease inhibitor shows promise for the diagnosis of protein-losing enteropathy.     

Fungal infections of the central nervous system in the dog and cat

Fungal infections of the central nervous system (CNS) in dogs and cats are uncommon. The purpose of this paper is to review the clinical signs, diagnostic tests, and therapeutic options of fungal infections of the CNS in the dog and cat. Clinical signs are dependent on lesion location and are often multifocal. Extraneural involvement is common. Antemortem diagnosis can be difficult and is definitively made via cytology, biopsy, or culture of an affected organ or cerebrospinal fluid (CSF). Magnetic resonance imaging can support a diagnosis and may assist in therapeutic decisions. Fungal serology can support a diagnosis when direct visualization of the organism is not possible. Long-term azole maintenance therapy is suggested to enhance survival and prevent relapse. Serial cerebrospinal fluid evaluation and magnetic resonance imaging may identify early relapse.     

Laboratory procedures for the diagnosis of gastrointestinal tract diseases of dogs and cats

An increasing number of laboratory tests are available for diagnosis of gastrointestinal tract diseases in dogs and cats. Use of these tests can lead to more accurate and rapid diagnoses. This review discusses laboratory tests, both new and old, and the role they currently play in the evaluation of animals presented with gastrointestinal problems. A minimum database helps assess the severity of the disorder, detect extra-gastrointestinal causes of problems and assists in formulating diagnostic and therapeutic plans.

Faecal examination remains one of the most important diagnostic procedures in the investigation of gastrointestinal problems. Zinc sulphate faecal flotation is an excellent routine screening technique for helminth and protozoal infections, including giardiasis. Rectal cytology can assist in the diagnosis of large bowel disorders. Interpretation of faecal immunodiagnostic tests is hampered by insufficient knowledge of test sensitivities and specificities. Routine faecal cultures are not warranted and faecal occult blood tests are rarely indicated.

Serum tests for gastric inflammation are now under development. The serum trypsin-like immunoreactivity test remains the gold standard technique for the diagnosis of exocrine pancreatic insufficiency. Breath hydrogen tests can be helpful in assessing the functional relevance of mild abnormalities in small-bowel biopsy specimens. Subnormal concentrations of serum cobalamin appear to be more specific indicators of gastrointestinal disease in cats than in dogs. Tests for small intestinal bacterial overgrowth remain controversial and assessment of gastrointestinal permeability has yet to prove its value in the diagnostic assessment of companion animals with gastrointestinal problems. Faecal alpha₁-protease inhibitor (α₁-PI) shows promise for the diagnosis of protein-losing enteropathy.     

Treatment of potassium balance disorders.

Potassium is the predominant intracellular cation and is critical for the maintenance of resting cellular membrane potential. Abnormalities of potassium balance can manifest as skeletal and cardiac muscle dysfunction. Abnormalities of potassium concentration in plasma can result from changes in external potassium balance (intake vs. excretion) or internal balance (intracellular to extracellular). Hyperkalemia can result from renal failure, uroperitoneum, or severe dehydration and acidosis in calves with diarrhea. Hypokalemia occurs due to reduced forage intake, when increased gastrointestinal losses occur as with diarrhea, due to increased renal losses as with metabolic alkalosis or exogenous corticosteroid administration which promote kaliuresis, or with redistribution of potassium into the intracellular compartment with alkalosis or in association with insulin-mediated glucose uptake. Aggressive intravenous and oral therapy are often necessary to correct potassium balance disorders, in addition to therapy aimed at correcting any underlying disorder contributing to the potassium imbalance.     

Detection of corn intrinsic and recombinant DNA fragments and Cry1Ab protein in the gastrointestinal contents of pigs fed genetically modified corn Bt11

Genetically modified corn has been approved as an animal feed in several countries, but information about the fate of genetically modified DNA and protein in vivo is insufficient. Genetically modified corn Bt11 is developed by inserting a recombinant DNA sequence encoding insecticidal Cry1Ab protein from Bacillus thuringiensis subsp. kurstaki. We examined the presence of corn intrinsic and recombinant cry1Ab gene by PCR, and the Cry1Ab protein by immunological tests in the gastrointestinal contents of five genetically modified corn Bt11-fed and five nongenetically modified corn-fed pigs. Fragments of corn zein (242 bp), invertase (226 bp) and of ribulose-1,5-bisphosphate carboxylase/ oxygenase genes (1,028 bp) were detected in the gastrointestinal contents of both Bt11 and nongenetically modified corn-fed pigs. Fragments of recombinant cry1Ab gene (110 bp and 437 bp) were detected in the gastrointestinal contents of the Bt11-fed pigs but not in the control pigs. Neither corn intrinsic nor cry1Ab gene fragments were detected in the peripheral blood by PCR. The gastrointestinal contents were positive for Cry1Ab protein by ELISA, immunochromatography, and immunoblot; however, these methods did not work for blood and precluded conclusions about any potential absorption of the protein. These results suggest that ingested corn DNA and Cry1Ab protein were not totally degraded in the gastrointestinal tract, as shown by their presence in a form detectable by PCR or immunological tests.     

Exocrine Pancreatic Insufficiency in the Dog: Historical Background,Diagnosis, and Treatment

This overview summarizes research performed during the last decades that has had an impact on the diagnosis and management of exocrine pancreatic insufficiency (EPI) in dogs. Pancreatic acinar atrophy is by far the most common cause for the maldigestion signs of canine EPI. The ability to diagnose pancreatic acinar atrophy in the subclinical phase before the development of total acinar atrophy and manifestation of clinical signs has offered new possibilities to study the pathogenesis of the disease. Diagnosis of exocrine pancreatic dysfunction is based on typical findings in clinical histories and clinical signs and is confirmed with pancreatic function tests. In recent years, the measurement of serum canine trypsin-like immunoreactivity has become the most commonly used pancreatic function test to diagnose canine EPI. Serum trypsin-like immunoreactivity measurement is species- and pancreas-specific. When clinical maldigestion signs of EPI appear, enzyme replacement therapy is indicated. Despite accurate enzyme supplementation, only a small portion of orally administered enzymes are delivered functionally intact into the small intestine. In dogs, the highest enzyme activity in the duodenum has been obtained with nonenteric-coated supplements: raw chopped pancreas or powdered enzymes. Aside from dietary enzyme supplements, dietary changes are often made to improve clinical response, but sometimes weight gain and stool quality remain suboptimal. Other medications for treatment of gastrointestinal tract signs are often used in such dogs with EPI. Antibiotics are the most common adjunctive medication. Of the antibiotics administered, tylosin is used in Finland almost exclusively.     

Lead toxicosis in cats-a review

Although the incidence of lead toxicosis in small animals continues to decrease, it remains a significant malady. We have reviewed the literature of the past 45 years, which revealed 70 cases involving cats. Sources, signs, diagnosis, pathology and treatment of feline lead toxicosis are reviewed. In 84% of these cases the source of lead was old paint usually from home renovation. The most common signs in cats are anorexia, vomiting, and seizures. The younger individuals seem more likely to show CNS signs. Since signs are often vague, lead toxicosis may be significantly under diagnosed in cats. The gold standard of diagnostic tests is blood lead concentration, although it does not necessarily correlate with total body burden of lead or with metabolic effects including clinical signs. Diagnostic tests including erythropoietic protoporphyrin (EPP), urine aminolevulinic acid, and others are discussed. Gross findings on necropsy are few and include a yellow-brown discoloration of the liver often with a nutmeg-like appearance. Histological examination may reveal pathognomonic inclusion bodies in liver and renal tissues. Characteristic histological changes in the CNS include neuronal necrosis and demyelination. Treatment of lead toxicosis in cats, as in any species, involves removing the exposure, decontaminating the individual and the environment, supportive care and chelation therapy. The most recently available chelator is succimer (meso 2,3-dimercaptosuccinic acid). Succimer given orally is well tolerated and has a wide margin of safety. A high index of suspicion of lead toxicosis is warranted in cats since they often present with vague and non-specific signs. With any consistent history owners need to be asked about home renovation. Early diagnosis and treatment affords a good prognosis.     

Proposal for rational antibacterial use in the diagnosis and treatment of dogs with chronic diarrhoea

Chronic diarrhoea is a frequent complaint in canine practice and the diagnostic path is often characterised by numerous diagnostic tests and stepwise empirical treatments, often applied before gastrointestinal endoscopy/mucosal biopsies. These include dietary interventions (novel protein, hydrolysed protein diet), parasiticides and still, in many cases, antibacterials. Indiscriminate use of antibacterial drugs risks detrimental consequences for both the individual patient (antimicrobial resistance, long-term disruption of intestinal bacterial populations, potential worsening of gastrointestinal signs) and the general public. For that reason, in this Perspective essay we advocate use of antibacterials only after histopathologic evaluation of gastrointestinal biopsies or, for those cases in which endoscopy is not possible, after other therapeutic trials, such as diet/pre-probiotics or anti-inflammatory drugs have proven unsuccessful. They should be reserved, after appropriate dietary trials, for those canine chronic diarrhoeic patients with signs of true primary infection (i.e. signs of systemic inflammatory response syndrome or evidence of adherent-invasive bacteria) that justify antibacterial use.     

Chronic pancreatitis with secondary diabetes mellitus treated by use of insulin in an adult California sea lion

CASE DESCRIPTION: A 21-year-old neutered male captive California sea lion developed chronic polyuria; polydipsia; polyphagia; accelerated development of existing cataracts; and frequent episodes of gastrointestinal upset including anorexia, signs of abdominal discomfort, diarrhea, and vomiting. CLINICAL FINDINGS: Chronic hypercholesterolemia, hypertriglyceridemia, hyperglycemia, and glucosuria were identified. During episodes of gastrointestinal abnormalities, transient hyperbilirubinemia and increased serum J-glutamyltransferase activities developed. Clinical findings strongly suggested chronic pancreatitis with secondary diabetes mellitus and intermittent cholestasis. Multiple diagnostic tests, including abdominal ultrasonography, serial hematologic and serum biochemical analyses, fecal examinations, urinalyses and bacteriologic culture of urine, measurement of serum fructosamine and insulin concentrations, and evaluation of thyroid and adrenal function, did not reveal any specific parasitic, endocrine, hepatic, or neoplastic etiologies. TREATMENT AND OUTCOME: For 1.5 years, the sea lion received once-daily administration of glargine insulin, gastrointestinal protectants, and a strict high-protein, low-fat diet. Daily monitoring of glucose regulation was achieved by training the sea lion to submit to blood and urine sampling. Glucose regulation ranged from fair to good, and clinical signs of diabetes mellitus lessened. Episodes of gastrointestinal upset still occurred, although the frequency and severity decreased. Ultimately, a severe episode developed, associated with diabetic ketoacidosis and sepsis, and the sea lion died. Severe fibrosing pancreatitis with exocrine and endocrine atrophy and abscesses arising from ectatic pancreatic ducts were found. Peripancreatic fibrosis caused stricture of the common bile duct, resulting in gallbladder distension without cholecystitis. CLINICAL RELEVANCE: Diabetes mellitus can occur secondary to chronic pancreatitis in California sea lions and insulin therapy should be considered.     

Fluid therapy in mature cattle.

Fluid therapy is practical and beneficial when properly administered to cattle. Mature cattle are more frequently alkalotic than acidotic, so nonalkalizing solutions are usually indicated. Exceptions include cattle with choke, carbohydrate engorgement, diabetes mellitus, and occasionally, renal disease, diarrhea, and fatty liver/ketosis. Many dehydrated cattle need supplemental potassium and calcium as well as sodium, chloride, and water. Intravenous administration is indicated in patients with obstructive gastrointestinal disease and those with severe dehydration. Oral or intraruminal administration is less expensive and, often, very effective.     

Fluid therapy in mature cattle

Fluid therapy is practical and beneficial when properly administered to cattle. Mature cattle are more frequently alkalotic than acidotic, so nonalkalizing solutions are usually indicated. Exceptions include cattle with choke, carbohydrate engorgement, diabetes mellitus, and, occasionally, renal disease, diarrhea, and fatty liver/ketosis. Most dehydrated cattle need supplemental potassium and calcium as well as sodium, chloride, and water. Intravenous administration is indicated in patients with obstructive gastrointestinal disease and those with severe dehydration. Oral or intraruminal administration is less expensive and often very effective.     

Procarbazine as adjunctive therapy for treatment of dogs with presumptive antemortem diagnosis of granulomatous meningoencephalomyelitis: 21 cases (1998-2004)

BACKGROUND: Granulomatous meningoencephalomyelitis (GME) is an idiopathic inflammatory disease of the central nervous system. Remission often is short-lived in dogs treated with glucocorticoids. Procarbazine is T cell-specific and crosses the blood-brain barrier. HYPOTHESIS: Dogs with presumptive antemortem diagnosis of GME given procarbazine as adjunctive therapy to prednisone will have improved long-term outcome compared with dogs given no treatment or glucocorticoids alone. ANIMALS: Two groups were studied: (1) Dogs with presumptive antemortem diagnosis of GME treated with procarbazine and prednisone (n = 21); (2) Dogs that had a histologic diagnosis of GME at postmortem examination and received no treatment (n = 11). METHODS: Dogs with presumptive GME treated with procarbazine were identified retrospectively from medical records of 2 veterinary referral hospitals. Selection criteria required all dogs have a neurologic examination, blood work, cerebrospinal fluid analysis, and brain imaging (MRI or CT). RESULTS: Median survival time for all dogs studied was 5.0 months. Median survival time for dogs treated with procarbazine was 14.0 months and for untreated dogs, 0.73 months. Treatment with procarbazine was significantly correlated with survival time (P < .001). Procarbazine was the only independent predictor of survival. Prednisone was reduced in dosage or discontinued in 17 dogs. Adverse reactions to procarbazine therapy included myelosuppression in 7 dogs and hemorrhagic gastroenteritis in 3 dogs. CONCLUSION: These data suggest that procarbazine treatment of presumptive GME may result in greater improved long-term outcome than has been previously reported for glucocorticoid treatment alone and may complement other immunomodulatory therapies. Procarbazine-treated dogs must be monitored for adverse reactions.     

The immunophenotype of peripheral blood lymphocytes in clinically healthy dogs and dogs with lymphoma in remission

Lymphoma is a common cancer of dogs that frequently is treated with chemotherapy or radiation therapy. Response to therapy is variable and currently available diagnostic tests do not reliably predict response to therapy. Treatment for lymphoma often results in lymphopenia, but it is unknown whether the changes in circulating lymphocytes result from generalized or specific reduction of lymphocytes. In this study, blood lymphocytes from 12 clinically healthy dogs, 10 dogs in remission because of treatment for B-cell lymphoma, and 8 dogs in remission from T-cell lymphoma were analyzed by flow cytometry by using a panel of 20 antibodies reactive with canine leukocyte antigens. Results identified similar lymphocyte parameters in treated dogs regardless of the type of lymphoma. Treated dogs had >50% reduction in blood lymphocyte concentration, and an absolute decrease in most subsets of lymphocytes. Both groups of treated dogs had relative increases in the proportion of CD3+, T-cell receptor (TCR)αβ+, and CD90+ lymphocytes, and a decreased proportion of CD45RA+ cells. In addition, dogs with T-cell lymphoma in remission had a significant increase in the proportion of CD49d+ lymphocytes. These findings were interpreted as representing likely suppression of lymphocyte regeneration by chemotherapy, with a relative increase in the proportion of memory over naïve lymphocytes. Lack of correlation with the T- or B-cell origin of the initial lymphoma suggested that, by using flow cytometric methods, residual circulating neoplastic cells could not be detected. However, the changes in the lymphocyte profile of dogs treated with chemotherapy may have relevance to their immunocompetence.