Diagnosis of Ehrlichia ewingii infection by PCR in a puppy from Ohio

Abstract: An 8‐week‐old, male, German Shepherd‐cross puppy found in southeastern Ohio was presented to The Ohio State University Veterinary Teaching Hospital for evaluation of lameness and lethargy. Fever and joint effusion involving multiple joints were identified on physical examination. Results of a CBC included mild anemia, mature neutrophilia, monocytosis, and hyperglobulinemia. Rickettsial morulae were identified within neutrophils in joint fluid and peripheral blood. Both initial and convalescent serum titers were negative for Ehrlichia sp.; however, PCR analysis was strongly positive for Ehrlichia ewingii. The patient's clinical signs resolved within several days of beginning doxycycline treatment. Resolution of infection was confirmed by negative PCR results after 18 days of treatment and again after 3 years. This is the first reported case of E. ewingii in Ohio. More importantly, this case demonstrates the importance of PCR in making a definitive diagnosis of tick‐borne disease and the potential pitfalls of relying on serologic testing alone in making a diagnosis.     

Equine Granulocytic Anaplasmosis

Equine granulocytic anaplasmosis (EGA) is a seasonal rickettsial disease of horses transmitted by Ixodes spp. ticks. The etiological agent is Anaplasma phagocytophilum, a coccobacillary gram-negative organism with a tropism for granulocytes. Clinical manifestations include fever, partial anorexia, depression, distal limb edema, petechiation, icterus, ataxia, and reluctance to move. Hematologic changes observed are thrombocytopenia, decreased packed-cell volume and marked leukopenia involving first lymphocytes and then granulocytes. Diagnosis is based on awareness of geographic area for infection, typical clinical signs, abnormal laboratory findings, and visualization of characteristic morulae in the cytoplasm of neutrophils and eosinophils in a peripheral blood smear. Treatment consists of the administration of tetracycline. The disease is being diagnosed with increasing frequency in the United States, Canada, Brazil and northern Europe.     

Clinical features of canine granulocytic anaplasmosis in 18 naturally infected dogs

BACKGROUND: Anaplasma phagocytophilum, the causative agent of canine granulocytic anaplasmosis (CGA), is a Gram-negative intracellular organism transmitted by ixodid ticks. Thus far, only a few clinical studies evaluating dogs with CGA have been published. OBJECTIVES: Evaluation of dogs naturally infected with A. phagocytophilum in which known co-infections were excluded. ANIMALS: Eighteen dogs with CGA. METHODS: Prospective study. The diagnosis of CGA was based on a positive PCR test result; dogs with co-infections were excluded. History, clinical findings, CBC, clinical biochemistry, infectious disease screening, diagnostic imaging, and the course of disease were evaluated. RESULTS: CGA was diagnosed based on a positive PCR test for A. phagocytophilum; 10 dogs also had morulae in neutrophils. Six of 18 dogs were seronegative to A. phagocytophilum, the others were seropositive. All dogs were acutely ill. The most common clinical findings were lethargy, inappetence, fever, and splenomegaly. Abnormal laboratory results included thrombocytopenia, anemia, lymphopenia, hypoalbuminemia, and abnormally high plasma alkaline phosphatase activity. In 6 of 10 dogs tested, the platelet-bound antibody test was positive; Coombs' test was negative in 9 dogs. All dogs were treated with doxycycline and recovered. PCR testing as well as blood smear analysis for morulae were negative in 14 tested dogs 2-8 weeks after beginning treatment. CONCLUSIONS AND CLINICAL IMPORTANCE: Clinical findings in dogs with CGA were nonspecific. Positive platelet-bound antibody test results suggest immune-mediated platelet destruction as an important pathogenic mechanism. With correct diagnosis and treatment, prognosis is good.     

Feline mesothelioma: case report and review of cytologic,immunocytochemical, histopathologic,and immunohistochemical findings

Mesotheliomas are uncommon neoplasms that arise from mesothelial cells in either the abdominal or thoracic cavities and are rarely diagnosed in cats. A 10-y-old spayed female domestic shorthair cat was presented to the Louisiana State University oncology service for evaluation of a large amount of abdominal effusion. Abdominal ultrasound identified a large mesenteric mass with numerous ill-defined nodules. An abdominocentesis was performed with cytologic and immunocytochemical findings consistent with a neoplastic effusion, with large clusters of epithelioid cells that exhibited strong cytoplasmic expression of pancytokeratin, vimentin, and Wilms tumor 1 antigens. Further testing was declined, and meloxicam was prescribed until the cat died 23 d after initial presentation. Upon postmortem examination, the omentum was contracted into a firm mass adhered to multiple organs and accompanied by numerous small white nodules throughout the abdominal cavity. On histopathology and immunohistochemistry, neoplastic cells were found throughout the abdominal cavity; 60–95% exhibited moderate-to-strong cytoplasmic immunoreactivity for cytokeratin, vimentin, and Wilms tumor 1 protein. The final diagnosis was an epithelioid mesothelioma. Our case illustrates the utility of cytology, immunocytochemistry, and its relation to histology and immunohistochemistry. We also reviewed the reported cases of feline mesothelioma.     

What is your diagnosis? Pleural fluid from a neonatal Thoroughbred filly with pneumonia

A 3-day-old filly was presented to the Cornell University Hospital for Animals with an umbilical hematoma and mild aspiration pneumonia. The foal underwent abdominal surgery for resection of the hematoma. Recovery was uneventful, but 3 days after surgery, the foal became progressively tachypneic. Imaging studies revealed bilateral pleural effusion and pleuropneumonia. Cytologic evaluation and bacterial culture of the pleural fluid from both sides of the chest revealed sterile exudates, consisting mostly of neutrophils, with fewer macrophages and lymphocytes. Pleural fluid macrophages contained variable amounts of purple-magenta globular material in their cytoplasm. A lighter colored granular precipitate was also seen throughout the background of the smears. Similar material was identified in a macrophage in a peripheral blood smear prepared 2 days after abdominal surgery. Large amounts of extracellular pink precipitate were also seen in the blood smear and persisted in the blood for 7 days after surgery. A protective lubricant, carboxymethylcellulose, had been instilled into the abdominal cavity during surgery to prevent intra-abdominal adhesions. The intracytoplasmic pigment within pleural fluid and blood macrophages and the extracellular precipitate in peripheral blood and pleural fluid smears was compatible with carboxymethylcellulose. The material was probably derived hematogenously and was considered an incidental finding. The pleuritis was attributed to exacerbation of the original aspiration pneumonia by the general anesthesia.     

Peritoneal effusion in cats: 65 cases (1981-1997)

OBJECTIVE: To determine signalment, diagnoses, presence of effusions in multiple sites, and outcome in cats with peritoneal effusion. DESIGN: Retrospective case series. ANIMALS: 65 cats. PROCEDURE: Medical records from 1981 to 1997 were reviewed to obtain information on cats with peritoneal effusion identified on physical examination, radiographs, abdominal ultrasonograms, or at necropsy. RESULTS: Conditions most commonly associated with peritoneal effusion in cats, in order of frequency, were cardiovascular disease, neoplasia, hepatic disease, renal disease, feline infectious peritonitis, peritonitis attributable to other causes, and urinary tract trauma. Dilated cardiomyopathy (DCM) was the most common disease associated with peritoneal effusion; however, DCM was diagnosed in most of these cats before taurine deficiency was found to be a primary cause of this form of cardiomyopathy in cats. Neoplasia was the most common cause after 1987. Right-sided congestive heart failure was the most commonly associated disorder in cats < 1 year old, whereas neoplastic disease was more common with increasing age. Most effusions were detected during the initial physical examination and were modified transudates. Peritoneal effusion was commonly accompanied by fluid accumulation elsewhere, particularly pleural effusion. The prognosis for a cat with abdominal effusion in this study was poor (mean survival time, 21 days; range, 1 to 350 days; median, 2.5 days). CLINICAL IMPLICATIONS: The primary differential diagnosis for peritoneal effusion in cats is neoplastic disease in older cats and right-sided heart failure in kittens. Diseases associated with peritoneal effusion generally have poor prognoses.     

Purulent pericarditis in a puppy

An eight-week-old female Cavalier King Charles spaniel was presented with tachypnoea, pyrexia and distended jugular veins following bite injuries to the chest and abdomen. Pericardial effusion was suspected from clinical, electrocardiographic and radiographic findings, and confirmed by echocardiography. Pericardiocentesis was carried out and the purulent fluid aspirated yielded Pasteurella multocida on culture. After drainage, broad spectrum antibiotic therapy was instituted and the puppy went on to make an excellent clinical recovery. Despite this, some hyperechoic areas of myocardium were observed on echocardiography and occasional ventricular premature complexes were seen on electrocardiography. These were probably the result of an associated myocarditis.     

Acute Pancreatitis in Cats With Hepatic Lipidosis

The purpose of this study was to characterize the incidence, clinical features, and prognosis of acute pancreatitis in cats with hepatic lipidosis. Of 13 cats histologically diagnosed with hepatic lipidosis between July 1988, and November 1989,5 (38%) were also histologically diagnosed with acute pancreatitis. In cats with hepatic lipidosis alone, the signalment, history, physical examination, and clinicopatho-logic findings were generally indistinguishable from those of cats with concurrent acute pancreatitis except that cats with acute pancreatitis were more likely to be cachectic and to have coagulation abnormalities. Hepatomegaly was seen on abdominal radiographs in both groups. Of the 5 cats with concurrent acute pancreatitis, abdominal ultrasonography detected 1 cat with a hypoechoic pancreas and 5 with peritoneal effusion; those abnormalities were not seen in cats without concurrent acute pancreatitis. Cats with concurrent acute pancreatitis had only a 20% recovery rate, compared with a 50% recovery rate in cats with hepatic lipidosis alone. We conclude that cats with hepatic lipidosis should be rigorously evaluated for concurrent acute pancreatitis because of 1) the rate of disease coincidence, 2) the inability of signalment, history, physical examination, and clinicopathologic findings to adequately distinguish between hepatic lipidosis and acute pancreatitis, 3) the worse prognosis associated with concurrent acute pancreatitis, and 4) the opposing nutritional strategies for hepatic lipidosis and acute pancreatitis. (Journal of Veterinary Internal Medicine 1993; 7:205–209. Copyright © 1993 by the American College of Veterinary Internal Medicine.)     

Fatal hepatic sarcocystosis in a puppy with eosinophilia and eosinophilic peritoneal effusion

A 3-month-old male Golden Retriever puppy was evaluated for lethargy and fever of 2-days duration. Results of a CBC and biochemical profile revealed marked eosinophilia (6.3 X 10(3)/microL; reference interval 0.1-1.2 X 10(3)/microL), moderate thrombocytopenia, and increased activities of alanine aminotransferase, aspartate aminotransferase, and creatine kinase. Hepatomegaly and peritoneal effusion were found using abdominal ultrasound. Peritoneal fluid analysis revealed eosinophilic inflammation (23,000 nucleated cells/microL with 88% eosinophils). Despite supportive treatment the puppy's condition deteriorated rapidly; euthanasia was requested, and a necropsy performed. Microscopically, there was marked necrosuppurative and eosinophilic hepatitis with vasculitis. Numerous hepatocytes contained protozoal organisms suspected to be Toxoplasma gondii or Neospora caninum. However, serum was negative for both T gondii and N caninum antibodies; polymerase chain reaction assay on hepatic tissue was negative for both organisms; and immunohistochemical evaluation of hepatic tissue using serum raised against T gondii, N caninum, and Sarcocystis neurona also was negative. Schizont morphology suggested that merozoites replicated by endopolygeny, forming rosettes around a central residual body. Transmission electron microscopy revealed that merozoites lacked rhoptries. These findings were consistent with a diagnosis of Sarcocystis canis, an apicomplexan parasite with an unknown life cycle.     

Duodenal perforation in a puppy with canine parvovirus infection

A 2-month-old puppy was brought to a veterinary hospital with diarrhea, vomiting, and anorexia. The test for canine parvovirus was positive, and she was hospitalized for supportive care. Her gastrointestinal symptoms initially improved; however, vomiting and lethargy developed again in the second week of hospitalization. Abdominal ultrasonography results were suspicious of a duodenal perforation. Cytology of the abdominal effusion confirmed septic peritonitis; therefore, emergency exploratory laparotomy was performed. The surgery was successful, and the puppy recovered fully. When symptoms recur or deteriorate in patients with parvoviral infection, surgically curable complications may be disregarded if supportive therapy is continued without additional investigative examinations. This report highlights the usefulness of abdominal ultrasound in conjunction with fluid cytology to identify subsequent complications when the clinical signs of parvovirus deteriorate.Key clinical message:This case report demonstrates duodenal perforation as a complication of parvoviral infection. Abdominal ultrasonography and peritoneal fluid cytology can be crucial for the early recognition of intestinal complications requiring immediate successful perioperative treatment.     

Hypereosinophilic paraneoplastic syndrome in a cat with intestinal T cell lymphosarcoma

A 10-year-old, neutered female, domestic shorthair cat was presented with a recent history of weight loss, polydipsia, diarrhoea and vomiting. On physical examination, intestinal thickening and mesenteric lymph node enlargement were apparent. Clinical investigations revealed peripheral blood eosinophilia, eosinophilic abdominal effusion and eosinophilic mesenteric lymphadenitis. There was a temporary response to treatment with glucocorticoids but signs progressed and the cat was euthanased. On histology, there was eosinophilic infiltration and fibroplasia of intestine and mesenteric lymph nodes. Large aggregates of neoplastic round cells in the intestine and lymph nodes were identified as T lymphocytes using immunohistochemistry. A diagnosis of intestinal T cell lymphosarcoma was made. This case demonstrates that hypereosinophilic paraneoplastic syndrome may occur in cats with lymphosarcoma. Eosinophil chemotaxis may be a response to the production of interleukin-5 by neoplastic lymphocytes.     

Evaluation of cytology in the diagnosis of acute canine monocytic ehrlichiosis (Ehrlichia canis): a comparison between five methods

The purpose of this study was the comparison of the diagnostic sensitivity between buffy coat (BC), peripheral blood (PB), lymph node (LN), bone marrow (BM) and short-term culture (P-D) cytology that has been based on the detection of Ehrlichia canis morulae, in the acute phase of canine monocytic ehrlichiosis (CME). Their cellular localization, total numbers and microscopic differentials were also investigated. The highest sensitivities were achieved after evaluating 1000 oil immersion fields (OIFs) in BC (66%) and an equal number in LN (60.9%) smears, separately or together (74%). The morulae were more often detected into lymphocytes than monocytes. The highest total number of morulae (n=143) were found in P-D smears. Finally, to avoid false positive diagnoses, platelets, lymphocytic azurophilic granules, lymphoglandular bodies and phagocytosed nuclear material should not be confused with the morulae.     

Babesia canis vogeli,Ehrlichia canis,and Anaplasma platys infection in a dog

A 12‐month‐old male neutered mixed breed dog was presented with a history of diarrhea, lethargy, emaciation, polydypsia, and sniffling. Physical examination findings included pale mucous membranes, increased heart and respiratory rates, and normal rectal temperature (38°C). Hematologic abnormalities included anemia and thrombocytopenia. Biochemical abnormalities included hypoalbuminemia, hyperbilirubinemia, and elevated ALP and ALT activities. A SNAP 4Dx test result was positive for Ehrlichia canis. Babesia canis vogeli organisms were found in the peripheral blood films, while morulae of E canis were not seen. Real‐time polymerase chain reaction testing confirmed the presence of both B c vogeli and E canis organisms, and also was positive for Anaplasma platys infection. The dog recovered following treatment with doxycycline and imidocarb dipropionate, with normal hematology and biochemical profiles.     

Seroprevalence and bacteremia [corrected] of Anaplasma phagocytophilum in cats from Bavaria and Lower Saxony (Germany)

Anaplasma (A.) phagocytophilum is a tick-transmitted obligate intracellular bacterium and has been identified in a wide range of mammalian species, causing febrile disease in some. Few reports show that it can also cause granulocytic anaplasmosis in cats. As data on the occurrence of A. phagocytophilum in cats from Germany is limited, a total of 326 serum and 306 EDTA-blood samples from cats from Germany were screened by direct (Giemsa-stained blood/buffy coat smears, real-time PCR) and indirect (IFAT) methods. Of 274 Giemsa-stained blood smears which could be evaluated none was positive for morulae, but one blood sample (< or =0.1%; 1/306) was positive for A. < or = phagocytophilum-DNA in PCR. Antibodies (cutoff > or = 1:64) were detected in 53 out of 326 samples (16.2%). Altogether, the results show a high seroprevalence rate of anti-A. phagocytophilum antibodies in cats in Germany while the low detection rate of this bacterial agent by direct methods is similar to those of other studies on A. phagocytophilum infections in cats.     

Large granular lymphoma in an FIV-positive and FeLV-negative cat

The clinical course of a feline leukaemia virus (FeLV)-negative and feline immunodeficiency virus (FIV)-positive cat affected with a large granular lymphocyte lymphoma is presented. Cyto-logical examination showed neoplastic cells in the pleural effusion and in two abdominal masses. Bone marrow and peripheral blood were moderately involved and chemotherapy was used to control the tumour. Cytochemistry, immunohis-tochemistry and ultrastructural studies were applied to define the cellular lineage; cytochemistry suggested a T-cell lineage.     

B‐cell lymphoma with Mott cell differentiation in two young adult dogs

Abstract: Two young adult dogs with gastrointestinal signs were each found to have an intra‐abdominal mass based on physical examination and diagnostic imaging. On exploratory laparotomy, small intestinal masses and mesenteric lymphadenopathy were found in both dogs; a liver mass was also found in dog 1. Cytologic and histologic examination of intestinal and liver masses and mesenteric lymph nodes revealed 2 distinct lymphoid cell populations: lymphoblasts and atypical Mott cells. With Romanowsky stains, the atypical Mott cells contained many discrete, clear to pale blue cytoplasmic inclusions consistent with Russell bodies that were positive by immunohistochemistry for IgM and CD79a in both dogs and for IgG in dog 2. The Mott cells and occasional lymphoblasts stained strongly positive with periodic acid‐Schiff. Using flow cytometric immunophenotyping in dog 1, 60% of peripheral blood mononuclear cells and 85% of cells in an affected lymph node were positive for CD21, CD79a, IgM, and MCH II, indicative of B‐cells. With electron microscopy, disorganized and dilated endoplasmic reticulum was seen in Mott cells in tumors from both dogs. Antigen receptor gene rearrangement analysis of lymph node and intestinal masses indicated a clonal B‐cell population. Based on cell morphology, tissue involvement, and evidence for clonal B‐cell proliferation, we diagnosed neoplasms involving Mott cells. To the authors' knowledge, this is the second report of Mott cell tumors or, more appropriately, B‐cell lymphoma with Mott cell differentiation, in dogs. More complete characterization of this neoplasm requires further investigation of additional cases. This lymphoproliferative disease should be considered as a differential diagnosis for canine gastrointestinal tumors.     

Surgical Treatment of Bile Peritonitis in 24 Dogs and 2 Cats: A Retrospective Study (1987–1994)

Objective — The purpose of this study was to determine the signalment, history, clinical signs, diagnosis, treatment, outcome, and factors affecting outcome of dogs and cats surgically treated for bile peritonitis. Study Design — Retrospective study. Animals or Sample Population — Twenty-four dogs and two cats surgically treated for bile peritonitis. Methods — The medical records of dogs and cats surgically treated for biliary effusions at the Ohio State University and Michigan State University between 1987 and 1994 were reviewed. Statistical analysis was performed to compare factors affecting outcome. Results — The cause of the biliary effusion was determined in 24 animals, and resulted from disruption of the biliary tract secondary to trauma (n = 13) or necrotizing cholecystitis (n = 11). Determination of the bilirubin concentration of the abdominal effusion was the only diagnostic test that was 100% effective in diagnosing bile leakage before surgical intervention. The bilirubin concentration of the effusion was consistently at least two times higher than the serum bilirubin concentration. Bacteriologic culture and sensitivity revealed that a septic, biliary effusion was usually associated with multiple types of gram-negative bacteria. The overall survival rate was 50% (13 of 26). The peripheral white blood cell count was significantly lower in survivors (mean 20,608/uL) compared with nonsurvivors (mean 35,712/uL). The immature neutrophil count was also significantly lower in survivors (mean 686/uL) than in nonsurvivors (4,852/uL). Only 27% (3 of 11) of the animals with a septic biliary effusion survived. In contrast, 100% (6 of 6) of the animals in which no bacteria were isolated from the abdominal effusion survived. Open abdominal drainage was not a successful treatment for 7 of 9 animals with septic biliary effusions. Survival was not significantly affected by the distribution of the peritonitis, cause of biliary effusion, or duration of clinical signs before surgical intervention. Conclusions — Patients with sterile biliary effusions have a much lower mortality rate than those with septic biliary effusions. The successful treatment of sterile biliary effusions does not require open abdominal drainage, and is not affected by the duration of the effusion. Clinical Relevance — This retrospective study provides information that may aid the surgeon in the diagnosis and treatment of bile peritonitis.     

Idiopathic or mesothelioma-related pericardial effusion: clinical findings and survival in 17 dogs studied retrospectively

This retrospective study compares the clinical signs and diagnostic findings of 17 canine patients with histopathological diagnoses of idiopathic pericardial effusion (IPE) or pericardial mesothelioma (MS) in order to identify differences in clinical findings or survival times that might aid in premortem differentiation of these disease conditions. Based on this series of cases, clinical signs, physical examination findings and results of non-invasive diagnostic testing are insufficient to differentiate MS from IPE with confidence unless a discrete pericardial or intrapericardial mass can be identified. Surgical biopsy may be misleading if large amounts of highly reactive and invasive mesothelial cells are seen. Recurrence of significant amounts of pleural effusion within 120 days of pericardiectomy may increase the likelihood that MS is the cause of pericardial effusion in cases in which other causes have been excluded. Survival longer than 120 days postpericardiectomy without chemotherapeutic intervention is associated with a decreased probability of the condition being MS.     

Multi-focal cerebral oligoastrocytoma in a puppy

A nine-month-old puppy was presented for investigation of seizures. Neurological deficits were found localising to the prosencephalon, mesencephalon and myelencephalon. Magnetic resonance imaging identified multiple, large lesions involving both cerebral hemispheres. Management with antiepileptic and immunomodulatory drugs was instituted; however, the seizures became progressively refractory and the puppy was euthanased. Histopathology following post-mortem examination found a mixed glial cell tumour with discrete areas where neoplastic cells appeared oligodendroglial or astrocytic. Primary intracranial neoplasia is a rare cause of neurological disease in young dogs. Moreover, this case is unusual in terms of both the mixed glial nature of the neoplasm and also its multi-focal distribution.     

Multisystemic Chromatolytic Neuronal Degeneration in Cairn Terriers: A Case With Generalized Cataplectic Episodes

Multisystemic chromatolytic neuronal degeneration, a newly recognized disease of Cairn Terriers, is described in a second affected North American puppy. In this puppy, the early onset of hind limb weakness at 11 weeks and rapid development of signs of diffuse CNS involvement were distinctive. Signs of cerebellar dysfunction were prominent, but bouts of cataplectic collapse in this puppy constituted the most distinguishing clinical feature. Although electroencephalograph (EEG) recordings lacked a true rapid eye movement (REM) pattern during cataplectic episodes, cervical electromyograph (EMG) potentials ceased or diminished, and imipramine injection was associated with arousal. Postmortem studies revealed that chromatolytic degeneration was very widespread, affecting many neuronal populations in the brain and spinal cord as well as neurons in sensory ganglia. Although the pattern of chromatolysis varied among affected perikarya, chromatolysis was consistently related to dispersion and loss of ribosomes. In this puppy, as opposed to six studied previously, thoracolumbar myelomalacia also occurred symmetrically in the dorsal horns and adjoining funicular white matter. The metabolic derangement underlying this chromatolytic neuronal degeneration and myelomalacia remains unknown.