Intracellular IFN-γ and IL-4 levels of CD4 + and CD8 + T cells in the peripheral blood of naturally infected (Leishmania infantum) symptomatic dogs before and following a 4-week treatment with miltefosine and allopurinol: a double-blinded, controlled and cross-sectional study.

BACKGROUND: Canine leishmaniosis (CanL) is a systemic disease caused by the protozoan parasite Leishmania infantum with a wide spectrum of clinical signs, with cutaneous, ocular, renal and lymphoreactive conditions prevailing in the clinical setting. The immune system plays a pivotal role in the evolution of Leishmania infection and its response to antileishmanial treatment. Cytokines are important immune response mediators that are released by activated lymphocytes and less so by other immunocytes. In dogs with leishmaniosis, IFN-γ and IL-4 have been recognized as the main activators of cellular and humoral immunity, respectively. The objective of this study was to investigate intracellular IL-4 and IFN-γ expression by CD4 + and CD8 + lymphocytes in the peripheral blood of symptomatic dogs before and after combined antileishmanial treatment with miltefosine and allopurinol. RESULTS: Postantileishmanial treatment CD4 + IL-4 + and CD8 + IL-4 + cell counts were significantly decreased, although no similar changes were observed in the comparisons made between the pre- and posttreatment CD4 + IFN-γ + and CD8 + IFN-γ + counts and ratios. CONCLUSION: The findings indicate that IL-4 production by T cells may facilitate the symptomatic phase of CanL, whereas IFN-γ production by CD4 + and CD8 + cells may indicate its negligible role in the evolution of natural CanL and perhaps the equivocal positive influence of antileishmanial treatment.

Norwegian or Crusted Sarcoptic Mange in Two Leishmanial Dogs.

Norwegian or crusted scabies (N/CS) is a rare skin disease with very few cases reported in the dog or the cat. Two adult, stray dogs were admitted in our clinic with a generalized, multifocal to diffuse and nonpruritic dermatitis that was characterized by severe crusting, scaling, and ulceration. In both instances, leishmaniosis and N/CS were diagnosed by immunofluorescent antibody test serology, lymph node cytology, and skin scrapings in which high numbers of Sarcoptes mites were found. The combination of miticidal and antileishmanial treatment, supported by topical treatment and nutritional support, resulted in the complete resolution of the skin lesions and spectacular improvement of the body condition in both cases. Dog 1 eventually died from end-stage kidney disease attributed to leishmaniosis-associated glomerulonephritis, whereas the also proteinuric dog 2 remains clinically healthy. The manifestation of the rare type of N/CS in these dogs could be attributed to cell-mediated immunosuppression, which was most likely induced by leishmaniosis and malnutrition. The necessity of searching for leishmaniosis in those scabietic cases, especially in the endemic areas of leishmaniosis, is strongly recommended.

Cutaneous Disease as Sole Clinical Manifestation of Protothecosis in a Boxer Dog.

Prototheca wickerhamii is ubiquitous, saprophytic achlorophyllous algae that cause opportunistic infections in the dog and cat and disseminated disease usually in immunocompromised animals. In this report an uncommon case of canine cutaneous protothecosis is presented. A 6-year-old female boxer was brought in with skin lesions that consisted of nodules and generalized footpad hyperkeratosis, depigmentation, and erosion. Cytology and histopathology showed pyogranulomatous inflammation along with organisms containing round sporangia with spherical sporangiospores. PCR and sequencing identified the causal organism as Prototheca wickerhamii. Therapy applied in this patient with either fluconazole alone or combination of amphotericin B and itraconazole proved effective only for footpad lesions but not for skin nodules. Systemic therapy seems to be ineffective for skin nodules, at least in chronic cases of canine cutaneous protothecosis. Although canine protothecosis usually presents with the disseminated form, cutaneous disease as sole clinical manifestation of the infection may also be witnessed.

Molecular identification of Bartonella species in dogs with leishmaniosis (leishmania infantum) with or without cytological evidence of arthritis.

Recent evidence suggest that Bartonella species may cause polyarthritis and lameness in dogs. Canine leishmaniosis (CanL) due to Leishmania infantum is a multi-systemic disease often occurring in association with arthritis. We hypothesized that concurrent Bartonella infection may be a contributing factor for the development of arthritis in dogs with CanL. Hence the primary objective of this study was to investigate the molecular prevalence of Bartonella spp. in dogs with naturally occurring CanL, with or without cytologically documented arthritis. Thirty-eight dogs with CanL (31 with neutrophilic arthritis and 7 without arthritis) were retrospectively studied. Seventy-four archived clinical specimens from these 38 dogs, including 33 blood samples, 19 bone marrow (BM) samples and synovial fluid (SF) aspirates from 22 dogs were tested for Bartonella spp. DNA using the Bartonella alpha proteobacteria growth medium (BAPGM) diagnostic platform. Overall, eight (21.1%) dogs were infected with one or two Bartonella species; however, Bartonella spp. infection was not associated with arthritis in dogs with CanL. Further prospective studies are warranted to determine if there is a correlation between Bartonella spp. infection and the development of arthritis in dogs with CanL.

Serial analysis of serum cardiac troponin I changes and correlation with clinical findings in 46 dogs with mitral valve disease.

BACKGROUND: Cardiac troponin I (cTnI) is a biomarker correlated with the severity of myocardial injury. It is hypothesized that serial assessment of cTnI could provide information about the disease progression in chronic heart failure. OBJECTIVE: The purpose of the study was to correlate serial serum cTnI concentrations with clinical scoring and select diagnostic imaging findings in dogs managed for mitral valve degeneration (MVD) for a period of 6 months. METHODS: Client-owned dogs with MVD were prospectively recruited for the study. The dogs were allocated into 3 groups (I, II, III) according to the severity of their clinical signs based on the classification suggested by the International Small Animal Cardiac Health Council. During the 6-month study period, serum specimens for biochemical testing were obtained biweekly, clinical progression and response to treatment were also evaluated biweekly, and radiographic reevaluation was performed every 2 months. RESULTS: A total of 46 dogs were evaluated. There was a marked decrease in cTnI values during the first 2 weeks after initial diagnosis, more pronounced in group III, and corresponding to the initiation of therapy and clinical stabilization of animals. Serum cTnI was significantly different (P < .05) among the 3 dog groups throughout the study period. The interaction between Group and Period was significant in group III, indicating an upward cTnI trend evident in severely affected animals. A positive correlation was demonstrated in all groups between serum cTnI and clinical scoring. CONCLUSION: Long-term management of MVD in dogs could benefit from the serial measurement of cTnI.

Serum canine pancreatic lipase immunoreactivity in experimentally induced and naturally occurring canine monocytic ehrlichiosis (Ehrlichia canis).

Ehrlichia canis infection causes multisystemic disease in dogs (canine monocytic ehrlichiosis, CME) which is associated with variable morbidity and mortality. Atypical clinical manifestations, including gastrointestinal signs, may occasionally occur in CME and approximately 10-15% of dogs are presented with historical or clinical evidence of vomiting, diarrhea, and/or abdominal discomfort. The objective of this study was to investigate if there are any alterations in serum canine pancreatic lipase immunoreactivity (cPLI) in dogs with experimentally induced or naturally occurring monocytic ehrlichiosis. Serum samples from 10 Beagle dogs experimentally infected with E. canis and two healthy uninfected Beagles were serially examined; samples from 20 naturally infected dogs (10 with non-myelosuppressive [NME] and 10 with myelosuppressive [ME] ehrlichiosis) were also examined at a given point in time (cross-sectional sampling). None of the experimentally infected Beagles showed gastrointestinal signs or increased cPLI concentrations prior to or following the artificial infection. Three naturally infected dogs with NME and one with ME demonstrated serum cPLI concentrations in the diagnostic range for pancreatitis (>400 µg/L) without showing gastrointestinal signs. The results of the present study indicated that 4/20 (20%) of dogs naturally infected with E. canis demonstrated increased serum cPLI concentrations consistent with mild and clinically inapparent pancreatitis.

Cutaneous involvement in canine leishmaniosis due to Leishmania infantum (syn. L. chagasi).

BACKGROUND: Canine leishmaniosis, a zoonotic infectious disease caused by Leishmania infantum (syn. L. chagasi), is fairly common in endemic areas, and skin lesions are the most frequent clinical manifestation. Despite the large volume of relevant scientific information, there is a relative paucity of up-to-date review articles on the cutaneous manifestations of the disease. HYPOTHESIS/OBJECTIVES: This review provides current information on the macroscopic and histological skin lesions of dogs with leishmaniosis and comments on their pathogenesis and definitive diagnosis. METHODS: Published articles have been thoroughly reviewed and the retrieved information has been used and supplemented as appropriate with the authors' clinical experience. RESULTS: The main clinicopathological presentations of canine leishmaniosis are exfoliative, ulcerative, nodular, sterile pustular and papular dermatitis and nodules at the site of parasite inoculation. Histopathology is usually dominated by mononuclear inflammatory infiltrates; immunohistochemical findings are variable, and Leishmania amastigotes are present in both diseased and normal-looking skin of dogs with leishmaniosis. Definitive diagnosis of the skin lesions in a dog with leishmaniosis is based on their macroscopic appearance, exclusion of main differentials, histopathology, demonstration of the parasite in the skin and complete response to antileishmanial treatment. CONCLUSIONS AND CLINICAL IMPORTANCE: Canine leishmaniosis due to L. infantum is characterized by diverse cutaneous manifestations that may reflect different host-parasite relationships. Furthermore, different types of skin lesions may occur, in various combinations, in the same dog. The definitive diagnosis of these lesions is based on clinical and laboratory examinations and on the response to antileishmanial treatment.

Markers of lipid peroxidation and α-tocopherol levels in the blood and neoplastic tissue of dogs with malignant mammary gland tumors.

BACKGROUND: Excess reactive oxygen species due to oxidative stress and the ensuing lipid peroxidation are believed to be involved in mammary gland tumor (MGT) pathogenesis. OBJECTIVE: The purpose of this study was to evaluate the extent of lipid peroxidation as evidenced by thiobarbituric acid-reactive substances (TBARS) formation, and the concentration of α-tocopherol as an inhibitor of lipid peroxidation, in blood and neoplastic tissue of dogs with malignant MGT. The correlation between inflammatory cell infiltration score and TBARS or α-tocopherol in MGT was also evaluated. METHODS: Sixteen intact female dogs with malignant MGT and 12 clinically healthy and age/weight-matched controls were included in the study. In all dogs, serum TBARS, α-tocopherol, total cholesterol, and triglycerides were measured. Tissue TBARS and α-tocopherol levels were determined in 1 cm(3) sized tissue samples collected from MGT and adjacent, ipsilateral, normal mammary gland tissue from the 16 affected dogs. The degree of inflammatory cell tumor infiltration was evaluated histologically. RESULTS: There were no statistically significant differences in serum levels of TBARS, α-tocopherol, total cholesterol, and triglycerides between dogs with and without malignant MGT. TBARS were significantly higher, whereas α-tocopherol was lower in neoplastic tissue when compared with normal mammary gland tissue. There was no correlation between TBARS or α-tocopherol concentration and the inflammatory cell infiltration score in neoplastic tissue. CONCLUSION: The increased level of TBARS suggests oxidative stress induction in canine malignant MGT. The origin of this phenomenon is not clear, as a potential oxidative burst could not be attributed to inflammatory cells infiltrating the tumors.

Effectiveness of a single application of 0·25% fipronil solution for the treatment of hirstiellosis in captive green iguanas (Iguana iguana): an open-label study.

Hirstiella spp. are common ectoparasites of captive green iguanas (Iguana iguana). Suggested treatments are empirical and some of them are of low efficacy and potentially toxic. The objective of this open-label study was to investigate the short-term efficacy and safety of a single application of 0·25% fipronil solution for the treatment of hirstiellosis. The skin of 50 green iguanas was thoroughly examined with the aid of bright light and magnifying lenses. A total of 21 iguanas were found to be infested, harbouring 1-24 mites (median: 5). All 35 mites collected from 17 iguanas were identified as Hirstiella sp. Both infested and non-infested lizards, sharing the same enclosure, were carefully wiped with 0·25% fipronil solution. The safety and the efficacy of the treatment were evaluated after 2 days in 47/50 (94%) and 7 days in 29/50 (58%) iguanas. Compared with pre-treatment levels, the parasitic load did not changed significantly on the second day but was significantly lower on day 7 (P = 0·006). No adverse reactions were noticed. Based on these results a single whole-body application of 0·25% fipronil solution can be considered a safe and effective treatment for the reduction of parasitic burden in captive green iguanas infested by Hirstiella sp. mites.

Efficacy of rifampicin in the treatment of experimental acute canine monocytic ehrlichiosis.

OBJECTIVES: To assess the efficacy of rifampicin in achieving clinical and haematological recovery and clearing infection in dogs with experimentally induced acute monocytic ehrlichiosis. METHODS: Five Ehrlichia canis-infected Beagle dogs were treated with rifampicin (10 mg/kg/24 h orally for 3 weeks), nine E. canis-infected dogs received no treatment (infected untreated dogs) and two dogs served as uninfected controls. Clinical score, platelet counts, immunofluorescent antibody titres and PCR detection of E. canis-specific DNA in blood, bone marrow and spleen aspirates were evaluated on post-inoculation days 21 (start of rifampicin), 42 (end of rifampicin) and 98 (end of the study). RESULTS: By day 21 post-inoculation, all infected dogs became clinically ill and thrombocytopenic, seroconverted and were PCR positive in at least one tissue. Clinical scores and antibody titres did not differ between the treated and infected untreated dogs throughout the study. The rifampicin-treated dogs experienced an earlier resolution of their thrombocytopenia (Kaplan-Meier survival plot, P=0.048), and the median platelet counts were significantly higher in the treated compared with the infected untreated dogs on post-inoculation days 42 (P=0.0233) and 98 (P=0.0195). At the end of the study, three treated and six untreated infected dogs remained PCR positive in one tissue each. CONCLUSIONS: The rifampicin treatment regimen applied in this study hastened haematological recovery, but was inconsistent in eliminating the acute E. canis infection.

Presumptive pure erythroid leukemia in a dog.

A 6.5-year-old, intact male Cocker Spaniel dog was referred with a history of depression and anorexia of 1-week duration. Mucosal pallor was prominent on physical examination. Complete blood cell count revealed pancytopenia and occasional blast cells. Bone marrow aspirate cytology indicated that individual particles were composed of approximately 60% hematopoietic cells and a monomorphic population of blast cells with perfectly round nuclei, consistent paranuclear clearing, and deeply basophilic cytoplasm devoid of granules dominating the marrow fields. The granulocytic lineage was severely decreased with a granulocytic-to-erythroid ratio of 0.15 and a blast cell percentage of at least 70% of all nucleated cells; the myeloblasts and monoblasts composed <5% of nonerythroid cells. Bone marrow cytology slides were submitted for immunocytochemical immunophenotyping using antibodies to myeloperoxidase, cluster of differentiation (CD)3, CD79a, CD11b, CD45, and CD34. The neoplastic cells did not express any of the antigens assessed. The combination of light microscopic cytomorphology and the immunophenotype were strongly suggestive of pure erythroid leukemia.

Serum acute phase proteins in dogs with symptomatic esophageal spirocercosis.

Canine spirocercosis (CS) is a helminthic infection caused by the nematode Spirocerca lupi. The clinical hallmark of the disease is esophageal dysphagia due to parasite-induced esophageal nodules. Currently, there is limited information on the involvement of serum acute phase proteins (APPs) in the symptomatic CS. The objective of this study was to investigate whether C-reactive protein (CRP), serum amyloid A (SAA), haptoglobin (Hp) and albumin are involved in CS, and if their concentrations measured on admission reflect the severity of benign esophageal lesions. Nineteen dogs with spontaneous symptomatic esophageal spirocercosis and 7 clinically healthy dogs were studied retrospectively. The most consistently increased APP in the symptomatic dogs was Hp (95% of the dogs), followed by CRP (68%). The SAA concentrations were infrequently increased (5% of the dogs), while albumin concentrations were decreased in 58% of the affected dogs. The dogs with spirocercosis had significantly higher median concentrations of Hp (p=0.0001) and CRP (p=0.02) compared to healthy dogs. Median albumin concentrations did not differ between the two groups of dogs. The median concentrations of Hp, CRP and albumin did not differ significantly between the dogs having a single or multiple esophageal nodules. The results of this study indicate that in symptomatic CS, Hp and CRP are significantly and consistently increased, while SAA and albumin may be of limited value as diagnostic markers. No association was established between the concentrations of Hp, CRP and albumin measured on admission and the number of esophageal nodules.

Serum cardiac troponin I concentrations in naturally occurring myelosuppressive and non-myelosuppressive canine monocytic ehrlichiosis.

The aim of this study was to determine whether serum cardiac troponin I (cTnI) concentration measured on admission was associated with the clinical severity of canine monocytic ehrlichiosis and was predictive of clinical outcome (death or survival) in dogs naturally infected with Ehrlichia canis. Serum cTnI concentration was compared among 22 dogs with non-myelosuppressive ehrlichiosis (NME), 22 dogs with myelosuppressive ehrlichiosis (ME) and 10 healthy dogs. Unlike healthy dogs, 45.5% NME and 59.1% ME dogs had increased cTnI concentrations. There was no difference in the frequency of cTnI increase or mean cTnI concentrations between the NME and ME groups, whereas mean cTnI concentration was significantly lower in healthy dogs. No association was established between cTnI concentration on admission and clinical outcome.

Dust mite species in the households of mite-sensitive dogs with atopic dermatitis.

BACKGROUND: The presence of important house dust and storage mite species in the microenvironment of atopic dogs has not been thoroughly investigated. OBJECTIVES: To compare the presence and population of five dust mite species (Dermatophagoides farinae, Dermatophagoides pteronyssinus, Acarus siro, Tyrophagus putrescentiae and Lepidoglyphus destructor) among households with mite-sensitive atopic dogs (Group A), households with clinically healthy dogs (Group B) and households without pets (Group C, n=25) in Greece. ANIMALS: Twenty mite-sensitive atopic dogs and 20 clinically healthy dogs. METHODS: Dust samples were collected with a vacuum cleaner from owners' mattresses (all groups) and from dogs' sleeping areas (Groups A and B) or living room couch (Group C), once every season of the year. Following dust flotation, mites were counted and identified. RESULTS: Dermatophagoides farinae was the most prevalent (60, 40 and 64% in Groups A, B and C, respectively), followed by D. pteronyssinus (45, 35 and 48%, respectively), whereas the three storage mites were found in fewer households. No major differences could be found between Groups A and B or between households with (Groups A and B) and without dogs (Group C) regarding the presence or numbers of the five dust mite species. CONCLUSIONS AND CLINICAL IMPORTANCE: The presence and population of five common house dust and storage mite species does not differ among Greek households with mite-sensitive atopic dogs, households with healthy dogs and households without pets.

Moxidectin efficacy in a goat herd with chronic and generalized sarcoptic mange.

A case of sarcoptic mange affecting almost all the animals of a dairy goat herd is described. This pruritic skin disease led progressively to high mortality and dramatic drop of milk yield. The lesions of the affected goats were typical of a chronic and generalized-diffuse sarcoptic mange. Diagnosis was confirmed by skin scrapings and histopathology in which many mites were demonstrated. All surviving goats were treated with injectable moxidectin solution 1% (CYDECTIN-Fort Dodge) at the dose of 0.2 mg/kg, applied every 15 days for four times, subcutaneously. Although pruritus had decreased soon after the first treatment, a satisfactory healing of cutaneous lesions was witnessed 6 weeks after the beginning of moxidectin trial. Parasitological cure was achieved in all affected animals by the end of the trial. In the four monthly followups, no evidence of sarcoptic mange could be found.

Cytologic patterns of lymphadenopathy in canine monocytic ehrlichiosis.

BACKGROUND: Recognition of different cytologic patterns in lymph nodes (LNs) from dogs with canine monocytic ehrlichiosis (CME) and noninfectious causes of lymphoid reactivity may have diagnostic utility. OBJECTIVES: The aims of the present study were to compare cytologic patterns in LNs of dogs with different phases of CME, to investigate the association of cytologic pattern and presence of Ehrlichia spp. morulae, and to compare patterns of lymphoid reactivity between dogs with CME and those with noninfectious causes of lymphoid hyperplasia. METHODS: Cytologic preparations of LNs from 35 dogs with nonmyelosuppressive CME (group A), 16 dogs with myelosuppressive CME (group B), 26 dogs with noninfectious diseases (group C), and 15 healthy dogs (group D) were evaluated. Percentages of lymphocyte types, plasma cells, macrophages, neutrophils, and eosinophils were determined. Samples from dogs in groups A and B were evaluated for the presence of morulae. RESULTS: Cytologic abnormalities in LNs were recorded in 54% of dogs in group A, 88% in group B, 39% in group C, and 0% in group D and were more frequent (P=.02) in dogs with myelosuppressive CME than those with nonmyelosuppressive CME. Plasma cell hyperplasia was more frequent in CME than in noninfectious diseases (P=.03). An association between the presence of cytologic abnormalities and morulae in group A dogs was not found. CONCLUSIONS: Dogs with myelosuppressive CME have more lymphoid cytologic abnormalities than dogs with nonmyelosuppressive CME. LN plasmacytosis is the major pattern of lymphadenopathy in dogs with CME and is found more frequently in dogs with CME than in dogs with noninfectious causes of lymphadenopathy.

Claw histopathology and parasitic load in natural cases of canine leishmaniosis associated with Leishmania infantum.

Histological lesions and the presence of Leishmania spp. amastigotes in claw tissues were investigated in 40 dogs with leishmaniosis, with (16/40--group A) or without (24/40--group B) generalized onychogryphosis. Following euthanasia, the entire third phalanx with intact claw was amputated, formalin fixed, decalcified in a formic acid solution, embedded in paraffin, sectioned longitudinally and stained with haematoxylin and eosin, and acid orcein-Giemsa. Nested polymerase chain reaction (PCR) was used for the detection of Leishmania amastigotes. Lichenoid mononuclear infiltration (all dogs in group A, 21 of 24 dogs in group B), basal keratinocyte vacuolation (nine of 16 dogs in group A, 15 of 24 dogs in group B) and dermoepidermal clefting (13 of 16 dogs in group A, 18 of 24 dogs in group B) were the most prominent histopathological findings. There was no difference in the frequency and severity of these lesions between the two groups. Leishmania amastigotes could not be visualized in the dermis of any of the H&E sections, but their presence was demonstrated by nested PCR in three of 16 dogs in group A and two of 24 dogs in group B. However, the frequency of positive nested PCRs was not significantly different between the two groups. In conclusion, claw histopathology in symptomatic dogs with leishmaniosis, either with or without onychogryphosis is mainly characterized by mononuclear lichenoid dermatitis with or without interface dermatitis and dermoepidermal clefting, and is not accompanied by substantial local parasitism.

Presence and density of domestic mites in the microenvironment of mite-sensitive dogs with atopic dermatitis.

This study was designed to investigate the presence and density of domestic mites (DMs) in households with atopic dogs sensitive to these mites (group A; n=20), in households with clinically healthy, nonatopic dogs (group B; n=20) and in households without pets (group C; n=25). Dust samples were vacuum-collected from the owner mattress (all groups) and dog sleeping area (groups A and B) or living room couch (group C) on four consecutive occasions, reflecting the four seasons of the year. DMs were found, at least once, in 19 of 20 (95%) group A, 13 of 20 (65%) group B and 21 of 25 (84%) group C households. DM numbers per gram of dust were 0-159 (median, 8.8), 0-302 (median, 3) and 0-1473 (median, 6.9) for group A, B and C, respectively. Dermatophagoides farinae predominated in all groups, since it was identified in 60% of group A, 40% of group B and 64% of group C households. Dermatophagoides pteronyssinus was found in 45%, 35% and 48% of households, in group A, B and C, respectively. No differences were found between households with (groups A and B) or without dogs (group C). When considering both sampling sites together, frequency of DM recovery was higher in group A than in group B (P=0.044). Also, both mite frequency (P=0.011) and density (P=0.015) in dog sleeping area were higher in group A than in group B. In conclusion, presence and density of DMs is higher in the microenvironment of mite-sensitive dogs with atopic dermatitis than in that of clinically healthy nonatopic dogs.

Factors affecting the occurrence, duration of hospitalization and final outcome in canine parvovirus infection.

The objectives of this matched case-control study in a veterinary teaching hospital were to investigate the influence of signalment and historical data on the odds of occurrence of canine parvovirus (CPV) enteritis and the potential usefulness of the clinical signs and clinicopathologic abnormalities recorded on admission as prognostic indicators of mean duration of hospitalization (DOH) and outcome of the disease. Ninety-four puppies with natural CPV enteritis and 188 age-matched controls were studied. The odds to develop CPV enteritis were higher in purebreds compared to mixed-breed puppies. Vomiting and depression at the time of admission were associated with a prolongation of DOH by 2 and 1.75 days, respectively. The lymphopenic and hypoalbuminemic dogs were hospitalized for 1.9 and 2.5 more days, respectively, compared to those without these abnormalities. The odds of non-survival were higher in those puppies with evidence of systemic inflammatory response syndrome (SIRS) at the time of admission.

Evaluation of a serum-based PCR assay for the diagnosis of canine monocytic ehrlichiosis.

The aim of this study was to estimate the relative diagnostic sensitivity and specificity of a polymerase chain reaction (PCR) assay in the serum of dogs with naturally occurring non-myelosuppressive canine monocytic ehrlichiosis (CME), and to investigate the association between PCR positivity and immunofluorescence antibody (IFA) titres for Ehrlichia canis. Serum samples obtained from 38 dogs with non-myelosuppressive CME and 12 healthy dogs were analyzed retrospectively. Each serum sample was analyzed in triplicate using an E. canis-specific nested PCR assay targeting a 389bp sequence of the 16S rRNA gene. E. canis DNA was amplified in 24 of 38 (63.1%) affected dogs; all samples from healthy dogs were negative. A high level of agreement was found among the PCR replicates (P<0.0001). Median IFA titre of the 24 PCR-positive dogs was significantly lower than that of the PCR-negative infected dogs (P=0.0029), indicating that E. canis DNA may circulate prior to the development of a high antibody titre. Serum-based PCR analysis is suggested for the early diagnosis of CME when whole blood samples are not available.