Progressive gammopathy and coagulopathy in a young English bulldog.

A restricted polyclonal or biclonal gammopathy resulting in bleeding tendencies was diagnosed in a young, neutered male English bulldog with concurrent splenomegaly, anemia, and severe elevations in IgM and, to a lesser degree, IgA immunoglobulins. There was a positive clinical response to treatment with prednisone and chlorambucil. This case bears similarity to a recently published syndrome of polyclonal gammopathy that is not neoplastic in origin in this breed. Key clinical message: The current case describes the management and clinical course of a recently described syndrome of polyclonal gammopathy in English bulldogs.

Gammapathie et coagulopathie progressives chez un jeune bouledogue Anglais. Une gammapathie polyclonale restreinte ou biclonale résultant en une tendance aux saignements fut diagnostiquée chez un jeune bouledogue Anglais mâle castré, avec une splénomégalie concomitante, de l'anémie et une augmentation sévère des immunoglobulines IgM et, à un degré moindre, des IgA. Une réponse clinique positive au traitement avec de la prednisone et du chlorambucil fut notée. Ce cas comporte des similarités avec un syndrome récemment décrit de gammapathie polyclonale qui ne serait pas d'origine néoplasique chez cette espèce.Message clinique clé :Le présent cas décrit la gestion et l'évolution clinique d'un syndrome récemment décrit de gammapathie polyclonale chez les bouledogues Anglais.(Traduit par Dr Serge Messier).

Clinical, haematological and biochemical findings in tigers infected by Leishmania infantum.

BACKGROUND: A large number of animal species are susceptible to Leishmania infantum (Kinetoplastida, Trypanosomatidae) in endemic areas, including domestic and wild felids such as tigers (Panthera tigris). Knowledge on the infection of this endangered species is still at its infancy, and therefore this study aims to identify clinical presentation and clinicopathological findings of tigers naturally infected by L. infantum. RESULTS: Tigers either L. infantum-positive (group A) or -negative (group B) were apparently healthy or presented visceral leishmaniasis unrelated conditions, except for one animal in which a large non-healing cutaneous lesion was observed. However, histological exam and immunohistochemistry carried out on the lesion excluded the presence of L. infantum amastigotes. Biochemical analysis showed that the average concentration of total proteins, globulins and haptoglobin were significantly higher (p < 0.01, p = 0.01 and p = 0.02, respectively), while the albumin/globulin ratio significantly lower (p = 0.05) in group A compared with group B. The biochemical alterations were partially confirmed by the serum protein electrophoresis results revealing a significant increase in the total protein value (p = 0.01) and hypergammaglobulinemia (p = 0.03) but an unmodified albumin/globulin ratio in group A. CONCLUSIONS: In this study tigers infected by L. infantum have shown to be mainly asymptomatic. The absence of clinical signs may lead veterinarians to overlook leishmaniasis in animals kept in captivity. Therefore, diagnostic and screening tests as serology should be part of routinely surveillance programs to be performed on tigers in zoological gardens located in endemic areas. Though only few protein-related laboratory abnormalities were recorded in infected animals, they could provide diagnostic clues for a first suspicion of L. infantum infection in tigers. Indeed, considering the high risk of zoonotic transmission in heavily frequented environment as zoos, a prompt diagnosis of L. infantum infection is of pivotal importance.

HYPERGAMMAGLOBULINEMIA AND MYELOMA IN FIVE TIGERS (PANTHERA TIGRIS): CLINICOPATHOLOGICAL FINDINGS.

Five adult tigers (Panthera tigris) presented with a range of clinical signs, including paresis (2/5), lameness (2/5), ataxia (3/5), anorexia (5/5), and lethargy (5/5). Each tiger demonstrated elevated plasma globulin levels (7.8-14.8 g/dl; [reference interval 2-5.1 g/dl]) on routine biochemistry, confirmed as a monoclonal gammopathy using protein electrophoresis. Serum gammaglobulin concentration ranged from 5 to 7.5 g/dl, or 45.1-63.4% of total protein concentration. Azotemia was present in three tigers. Diagnostics and management varied with the presenting signs but included magnetic resonance imaging, radiography, chemotherapy, supportive care, and euthanasia. In each case, necropsy revealed a neoplastic plasma cell proliferation in the bone marrow and one or more extramedullary sites. Lytic lesions in the thoraco-lumbar spine were found in three animals, and one lesion was associated with spinal cord compression. Splenomegaly was present in 4/5 cases. Histopathology confirmed a plasma cell neoplasm in each case, and immunohistochemistry staining with multiple myeloma oncogene 1 (MUM1) was positive in each case. CD20 staining was performed in two cases and was positive in one. CD3 staining was performed in the same two cases, and was negative in each. Based on the clinical, gross, microscopic, and immunohistochemical findings, myeloma was diagnosed in all five tigers.

Monoclonal hyperglobulinemia in lymphosarcoma in a cockatiel (Nymphicus hollandicus) and a blue and gold macaw (Ara ararauna).

A 15-yr-old cockatiel (Nymphicus hollandicus) presented with decreased activity and appetite. A presumptive diagnosis of lymphoproliferative disease was made because of a monoclonal band observed on plasma protein electrophoresis. The owner declined further diagnostics or treatment, and the bird died 2 yr later. In a second case, a 4-yr-old blue and gold macaw (Ara ararauna) presented with lethargy and anorexia. Despite supportive treatment, the bird died 7 days later. Blood studies also demonstrated a monoclonal band on plasma protein electrophoresis. The histologic review of tissues from both birds found extensive neoplastic lymphoplasmocytic cells in the liver and supported a diagnosis of lymphosarcoma.

Comparative study of the plasma globulin level, CD21(-) B-cell counts and FOXP3 mRNA expression level in CD4(+) T-cells for different clinical stages of feline immunodeficiency virus infected cats.

Feline immunodeficiency virus (FIV) infection leads to hypergammaglobulinemia through mechanisms that remain poorly understood. We investigated changes in plasma globulin level, B cells, and T cells with progression of the clinical stage of FIV-infected cats. We classified FIV-infected cats into the stage of Asymptomatic carrier (AC) and AIDS-related complex (ARC) based on the clinical symptoms, and measured the plasma globulin level, the CD4(+) T-cell counts, and analyzed surface markers of B cells. We investigated the relationship between the plasma globulin level and regulatory T cells (Tregs) using the Forkhead box P3 (FOXP3) mRNA expression level. In FIV-infected cats, the plasma globulin level and the surface immunoglobulin (sIg)(+) CD21(-) B-cell counts were increased, whereas the CD4(+) T-cell counts were decreased compared with specific-pathogen free (SPF) cats. The mRNA expression of Blimp-1 (master gene of plasma cells) was increased in peripheral blood, and the FOXP3 mRNA expression level was decreased in CD4(+) T-cells. These immunological changes were marked in the ARC stage. These data indicate that the decrease of Tregs and the increase of plasma cells lead to hypergammaglobulinemia.

Use of recombinant interferon omega in feline retrovirosis: from theory to practice.

Type-I interferons (IFNs) are cytokines that have non-specific antiviral activity, participating mostly in innate defense mechanisms. Their administration has been proposed to treat several viral and immunomediated diseases as an immunomodulatory therapy. Due to its availability, recombinant human interferon-alpha (rHuIFN-α) has been studied in relation to feline retrovirosis, both in vitro and in vivo. However, IFNs are species-specific and antibodies have been shown to develop in response to the high rHuIFN-α doses necessary for an effective therapy. A recombinant feline IFN has been developed, which has been characterized as interferon-omega (rFeIFN-ω), designed to overcome these problems. Nonetheless, very few studies have been undertaken to evaluate its efficacy in cats naturally infected with FIV or FeLV. In an initial study, we here demonstrated that rFeIFN-ω can dramatically improve the clinical condition of infected cats, and induce improvement of hematologic parameters. Minor changes or no change was observed for hypergammaglobulinemia, CD4/CD8 ratio, proviral load, viremia and RT activity, suggesting that the overall effect of IFN was on innate immunity. More studies are needed in order to better understand its in vivo mechanisms.

Hypergammaglobulinemia in an SIV-infected rhesus macaque with a B-cell neoplasm with plasma cell differentiation.

An SIV-infected rhesus macaque presented with anemia, hypercalcemia, and hyperglobulinemia. Neoplastic round cells with plasma cell morphology infiltrated multiple organs and stained immunohistochemically positive for CD45, MUM1/IRF4, CD138, VS38C, and Kappa light chain and variably positive for CD20 and CD79a, consistent with a B-cell neoplasm with plasma cell differentiation.

Facial paralysis and lymphocytic facial neuritis in a rhesus macaque (Macaca mulatta) positive for simian retrovirus type D2.

Simian retrovirus type D (SRVD) is a naturally occurring betaretrovirus in nonhuman primates of the genus Macaca. Infection can lead to a variety of clinical, hematologic, and histopathologic abnormalities. We report an unusual clinical presentation of facial paralysis and histologic lymphocytic neuritis in an SRVD type 2 (SRVD2)-infected rhesus macaque (Macaca mulatta) with a catheter-associated vena caval thrombus, anemia, thrombocytopenia, and multisystemic lymphoid hyperplasia. At initial presentation, a right atrial mass was detected by echocardiography. The macaque was clinically asymptomatic but had persistent anemia, thrombocytopenia, hyperglobulinemia, and later neutropenia. It was seropositive for SRV and PCR-positive for SRVD 2. Approximately 1 mo after initial presentation, the macaque developed right facial paralysis and was euthanized. Histologic lesions included lymphoplasmacytic aggregates affecting multiple organs, consistent with SRV-related lymphoid hyperplasia. The right facial nerve showed lymphoplasmacytic inflammation. The nerve itself was negative immunohistochemically for SRV antigen, but antigen was present infrequently in pericapillary lymphoid cells within the facial nerve and abundantly within lymphoid aggregates in the adjacent parotid salivary gland, bone marrow, and soft tissue. Known neurotropic viruses could not be identified. Given the widespread inflammation in this macaque, particularly in the area surrounding the facial nerve, lymphocytic neuritis and facial paralysis likely were an indirect effect of SRV infection due to local extension of SRV-related inflammation in the surrounding tissue.

Sporotrichosis: the main differential diagnosis with tegumentary leishmaniosis in dogs from Rio de Janeiro, Brazil.

Seventy-four dogs from the State of Rio de Janeiro with ulcerated cutaneous lesions were submitted to clinical, dermatological, parasitological, mycological, histopathological and cytopathological exams, a leishmanin skin test, an indirect immunofluorescence (IIF) test for leishmaniosis, and nonspecific laboratory tests such as blood count and serum biochemistry. Sporothrix schenckii was isolated from 41 dogs and Leishmania (Viannia) braziliensis was isolated from 33 animals. Most dogs with sporotrichosis were from the municipality of Rio de Janeiro (53.7%) and presented ulcerated cutaneous lesions on the head (68.3%). Laboratory alterations in these animals included anemia (58.5%), hypoalbuminemia (83%) and hyperglobulinemia (75.6%). Histopathology revealed the predominance of a chronic granulomatous inflammatory infiltrate (70.7%), and yeast-like structures were detected in 17% of the histopathological exams and in 32% of the cytological exams. Three of 41 dogs with sporotrichosis were seropositive by IIF for leishmaniosis and 2 of 20 animals tested within this group had a positive leishmanin skin test. Similarly, most of the 33 dogs with leishmaniosis were from the municipality of Rio de Janeiro (69.7%) and had ulcerated cutaneous lesions on the head (84.8%). Laboratory alterations in these animals included anemia (66.7%), hypoalbuminemia (100%) and hyperglobulinemia (91%). Histopathology showed the predominance of a chronic granulomatous inflammatory infiltrate (63.6%) and amastigote forms were detected in 30.3% of the histopathological exams and in 31.8% of the 22 cytological exams performed. About 72.7% of the dogs were seropositive by IIF and five of seven animals had a positive skin test. Due to the clinical similarities, histopathological and nonspecific laboratory results similarities, the serological and skin tests for leishmaniosis positive in dogs with sporotrichosis, and the overlapping endemic areas in Rio de Janeiro, the differential diagnosis between the two diseases requires the demonstration of their respective etiological agents.

Canine sporotrichosis in Rio de Janeiro, Brazil: clinical presentation, laboratory diagnosis and therapeutic response in 44 cases (1998-2003).

A sporotichosis epidemic involving forty-four dogs in the Metropolitan area of Rio de Janeiro is described. Solitary skin lesions were noted in 18 dogs (40.9%), 2-4 such lesions were observed in 17 animals (38.6%), and nine (20.5%) animals had five or more lesions. Twenty-five (56.8%) animals had single ulcerated skin lesions on the nose and nine (20.5%) showed nasal mucosal involvement (three of which also has a skin lesion). Respiratory symptoms were observed in 17 (38.6%) dogs and were found to be the most common extracutaneous signs of infection. Anemia, leukocytosis with neutrophilia, hypoalbuminemia and hyperglobulinemia were the most frequent hematological abnormalities. Histopathological analysis of skin biopsies in most cases revealed granulomatous reactions characterized by histiocytic hyperplasia and neutrophil infiltration. Yeast-like cells were observed in seven (16.7%) of 42 dogs examined histologically. During the study, eight (18.2%) animals were lost to follow-up and three (6.8%) were submitted to euthanasia. Of the remaining 33 dogs, five (15.2%) presented spontaneous regression of the lesions, 26 (78.8%) were cured after treatment, and two (6%) continue to be treated. The present cases indicate that many dogs with sporotrichosis respond well to treatment and in a few dogs, the disease may be self-limiting.

Clinical and serological aspects of visceral leishmaniasis in northeast Brazilian dogs naturally infected with Leishmania chagasi.

Human visceral leishmaniasis is endemic in the northeast of Brazil, where the domestic dog is an important parasite reservoir in the infectious cycle of Leishmania chagasi. In this study, we evaluated the clinical signs of canine visceral leishmaniasis (CVL), serum protein profile and the antileishmanial IgG antibody production in 86 dogs living in northeast endemic areas of leishmaniasis. Thirty dogs from a leishmaniasis-free area were used as a control group. The major clinical signs of CVL seen were emaciation and skin ulcers (80%), followed by onychogryphosis and conjunctivitis (73%). Depilation was observed in 60% of animals while lymphadenomegaly, splenomegaly, liver enlargement or kidney involvement was less frequent (< or =20%). VL seropositive dogs presented with serum hyperproteinemia, hypoalbuminemia, hypergammaglobulinemia and decreased albumin/globulin ratio. A lower sensitivity and higher specificity was observed for promastigote indirect fluorescent antibody test (IFAT) (83 and 100%, respectively) compared with enzyme-linked immunosorbent assay (ELISA) (94 and 90%), which uses a crude extract of Leishmania. There was a positive correlation between IFAT and ELISA titers of antileishmanial IgG antibodies (Spearman test, P < 0.05), which was augmented in CVL dogs. This study found that the determination of serum protein, A/G ratio and the use of two different leishmanial serological tests like IFAT and ELISA are essential in CVL screening.

Clinical and pathological features of the Porcine Ulcerative Dermatitis Syndrome (PUDS).

The nomenclature of ulcerative dermatitis as used in literature is somehow confusing because on the one hand this skin disorder is associated with bacterial growth and on the other hand it is a synonym for a chronic sporadic disease of adult sows with unknown aetiology. Thus, we propose the terminus 'Porcine Ulcerative Dermatitis Syndrome (PUDS)' for the latter to distinguish between these two disease complexes. This syndrome could be identified by clinical and pathological examinations in six sows, that were submitted to the clinic. Epidermal ulcers could be found nearly all over the body, but teats were always spared. Haematological examination showed a slight anaemia but physiological leucocyte counts. However, lymphopenia (x = 44.8%), granulocytosis (x = 42.0%) and an increased number of monocytes (x = 13.1%) could be found. Histologically, a lymphoplasmacytic and granulohistiocytic infiltration in the corium was most prominent. In some cases, a moderate leucocytoclastic vasculitis and perivasculitis could be seen at the dermo-epidermal border. Additionally, a multifocal interstitial nephritis with lymphoplasmacytic infiltration was a prominent feature in all animals. Participation of an immune complex associated disorder can be assumed when regarding histological findings as skin lesions in combination with glomerulonephritis are a common feature of such diseases. Also, IgG levels were elevated two- to fourfold in all affected sows when compared with healthy control pigs. This supports the hypothesis that not only T cells, as shown previously, but also the humoral branch of the immune system is involved in the aetiology of PUDS.

Hypercalcemia in two Amazon parrots with malignant lymphoma.

Malignant lymphoma is a common malignancy in birds. Paraneoplastic syndromes, which are commonly observed in domestic animals, have not been reported in association with lymphoma in birds. Hypercalcemia and hyperglobulinemia were found on plasma chemistry in two Amazon parrots, which were presented with aspecific symptoms. In both cases radiography and ultrasound demonstrated signs of hepatomegaly, which proved to be due to malignant lymphoma on postmortem examination. The hypercalcemia was found to be most consistent with a paraneoplastic effect of the malignant lymphoma in these birds. The exact origin of the hyperglobulinemia remains unclear.

Analysis of serum cytokine profile in a holstein heifer with leukocyte adhesion deficiency which survived for long period.

Serum cytokine levels and their expression of mRNA on neutrophils from a bone marrow (BM) transplanted heifer with leukocyte adhesion deficiency were evaluated. The clinical condition of the affected heifer was relatively stable after BM-transplantation. Persistent hyper gamma-globulinemia and increased serum immunoglobulin G (IgG) concentrations were monitored longitudinally. The concentration of interleukin (IL)-1beta in serum from the affected heifer ranged from 15.8 to 321.7 ng/ml, and maximum concentration occurred at the time which coincided with peak IL-6. Serum levels of IL-6 ranged from 0.32 to 27.9 ng/m l, and they appeared to be associated with the increment of serum IgG in the affected heifer. mRNAs for IL-1beta, IL-6, IL-8 and granulocyte and macrophage colony stimulating factor (GM-CSF) were increased in neutrophils from the affected heifer compared to controls. Persistent hyper gamma-globulinemia of the affected heifer appeared to be associated with enhanced mRNA expression for IL-6 and its serum levels. These findings suggest that humoral immunity of the affected heifer is activated and the production of neutrophils appears to be enhanced under the incapability of beta(2) integrin-mediated functions of phagocytic cells.

Biclonal gammopathy associated with immunoglobulin A in a dog with multiple myeloma.

A 10-year-old neutered male Airedale Terrier was evaluated for inappetance, weight loss, and lameness. Multiple myeloma was diagnosed based on bone marrow plasmacytosis, multiple lytic bone lesions, and hyperglobulinemia with a clonal gammopathy on serum protein electrophoresis. Splenic plasmacytosis, and retinal lesions consistent with hyperviscosity syndrome also were found. Temporary responses to 2 different chemotherapy protocols (melphalan and prednisone, and cyclophosphamide and prednisone) were seen, with remission of clinical signs and a decrease in the biclonal gammopathy but no resolution of the splenic mass. Eventual return of clinical signs led to euthanasia at 175 days postdiagnosis. Necropsy examination confirmed multiple myeloma involving bone marrow and spleen, and glomerulonephritis. An immunoglobulin-A (IgA) gammopathy was demonstrated by immunoelectrophoresis; biclonality was ascertained by immunofixation electrophoresis. The clonal components consisted of intact Ig with heavy chain of the alpha class and light chain of an undetermined class. To our knowledge, this is the first report of undimerized biclonal gammopathy in a dog caused by a single heavy chain class involving IgA.

Extramedullary plasmacytoma in a horse with ptyalism and dysphagia.

A Clydesdale mare was examined for weight loss, inappetence, ptyalism, and dysphagia. The main abnormality revealed by serum biochemistry was a marked hyperglobulinemia, and protein electrophoresis revealed a monoclonal gammopathy in the gamma region. The urine was positive for Bence Jones proteins. These findings suggested a plasma cell tumor. The neoplasm could not be located with extensive antemortem examination. At postmortem, neoplastic cells morphologically compatible with plasma cells and positive for equine IgG with imunoperoxidase staining infiltrated the pericardium, mediastinal stromal tissues, adrenal glands, meninges, atrioventricular valves, aorta, abdominal and thoracic fat, and nerves, including the trigeminal nerve. The neoplastic cells invading the cranial nerves were responsible for many of the presenting signs.

Systemic kappaAL amyloidosis associated with bovine leukocyte adhesion deficiency.

Histopathologic and immunohistochemical examinations were conducted on a 5-year-old Holstein-Friesian cow with systemic kappaAL amyloidosis associated with bovine leukocyte adhesion deficiency. Amyloid deposits were present in the perivascular and intercellular spaces of the visceral organs, such as the liver, kidneys, pancreas, adrenal glands, and upper alimentary tract. Amyloid was stained positively with Congo red with or without 5% potassium permanganate pretreatment and had green birefringence observed under polarized light. Immunohistochemically, amyloid reacted strongly against anti-bovine IgG (H+L) and anti-bovine kappa-light chain and reacted weakly against bovine X-light chain antibodies but was negative for anti-human amyloid AA antibody. This is the first description of AL amyloidosis immunohistochemically related to immunoglobulin kappa-light chains of precursor protein in cattle.

Hyperglobulinemia and lymphocyte subset changes in naturally infected, inapparent carriers of equine infectious anemia virus.

OBJECTIVE: To determine blood protein concentration, immunoglobulin concentration, and lymphocyte profiles in equine infectious anemia virus (EIAV) seropositive, naturally infected horses without clinical signs of disease. ANIMALS: 26 clinically normal seropositive horses, 6 febrile ponies with experimentally induced EIA, and 52 clinically normal seronegative horses and ponies. PROCEDURE: Serum and EDTA-anticoagulated blood were obtained from all horses and ponies, and total serum protein and albumin concentrations, immunoglobulin concentrations, and blood lymphocyte subset counts were determined. RESULTS: Compared with seronegative horses, EIAV seropositive inapparent carrier horses had no significant difference in serum reverse transcriptase activity, PCV, or platelet count. Inapparent carrier horses had increased plasma total solids and serum globulin concentrations and decreased serum albumin concentration and albumin-to-globulin ratio. Total serum immunoglobulin and serum IgM concentrations were increased. Inapparent carrier horses had significantly decreased percentages of CD5+ and CD4+ blood lymphocytes. CONCLUSIONS: Serum protein and lymphocyte subset changes in EIAV-infected inapparent carrier horses are consistent with immune activation or chronic inflammation, both of which may, in part, be the result of virus-induced polyclonal B-cell activation. CLINICAL RELEVANCE: EIAV seropositive horses have immune-related abnormalities consistent with ongoing viral activity regardless of the duration they have been infected, even when the usual signs of disease (anemia, fever, weight loss) are not apparent.