Cytokines as immunological markers for systemic inflammation in dogs with pyometra.

Pyometra is a disease in dogs caused by bacterial infection of the uterus and resulting in SIRS (systemic inflammatory response syndrome) in nearly 6 of 10 cases. Clinical diagnostic criteria for SIRS are relatively unspecific, and biomarkers for the diagnosis of pyometra and SIRS in dogs are needed. Serum samples from 32 dogs were used in this study and grouped into dogs with pyometra and SIRS, dogs with pyometra without SIRS and healthy controls. The serum concentrations of IFN-γ, IL-4, IL-6, IL-7, IL-8, IL-10, IL-15, IL-18 and TNF-α were measured using multiplex analyses. The serum concentrations of CRP (C-reactive protein) were determined using sandwich ELISA. IL-7, IL-8, IL-15, IL-18 and TNF-α were detected in >94% of samples. IL-10 was detected in 28% of samples, and IL-4, IL-6 and IFN-γ were undetectable. Higher serum concentrations of IL-7 (p < 0.05) were detected in SIRS-positive dogs with pyometra (n = 13) as compared with healthy controls (n = 11). The concentrations of IL-8 were higher in SIRS-positive dogs with pyometra compared to the SIRS-negative group (n = 8; p < 0.05). Positive correlations of IL-15 with IL-18 (p < 0.0001) and with the concentrations of IL-7 (p < 0.0001 for both) were found, although there was no significant difference between groups. Furthermore, IL-15 correlated with concentrations of CRP (p < 0.05), which were higher in dogs with pyometra compared to controls (p < 0.0001). Our data suggest a role of several cytokines in the development of a systemic disease in dogs with pyometra and a possible diagnostic value for serum CRP, IL-7, IL-15 and IL-18 in canine SIRS caused by pyometra.

Plasma PGF 2 alpha metabolite levels in cats with uterine disease.

Uterine disease induces PGF(2 alpha) increase in many animal species, which can be measured by the metabolite 15-keto-(13,14)-dihydro-PGF(2 alpha) (PGFM). Plasma PGFM levels are associated with severity of the uterine disease and presence of systemic inflammatory response syndrome (SIRS) in dogs. The objectives in this study were to investigate PGFM levels, presence of SIRS, and clinical and laboratory parameters in female cats as possible indicators for severity of uterine disease. In total, 7 female cats with pyometra, 2 with mucometra, 7 with cystic endometrial hyperplasia (CEH), and 14 healthy control cats were included. Physical examination, ovariohysterectomy, and histopathology were performed, laboratory parameters were analyzed, and PGFM levels were analyzed by radioimmunoassay. Analysis of variance, Fisher's exact test, Student's t-test and Pearson's product moment correlation coefficient were used for data analysis. In cats with pyometra, mean PGFM levels were increased (21.1 nmol L(-1)) but were decreased in cats with CEH (0.4 nmol L(-1)) compared with control cats (0.6 nmol L(-1)). In cats with mucometra, the mean PGFM level was 8.8 nmol L(-1). Systemic inflammatory response syndrome was present in 6 (85%) cats with pyometra, 1 cat with mucometra, and 1 cat with CEH. Hospitalization length was negatively correlated with albumin and positively correlated with total white blood cell count (WBC), neutrophils, band neutrophils (BN), percentage BN (PBN), and monocytes. Pyometra and mucometra were associated with increased plasma levels of PGFM. The parameters albumin, WBC, neutrophils, BN, PBN, and monocytes may be useful to determine morbidity as measured by hospitalization length.

Molecular evidence for persistence of Anaplasma phagocytophilum in the absence of clinical abnormalities in horses after recovery from acute experimental infection.

BACKGROUND: Anaplasma phagocytophilum infects several mammalian species, and can persist in sheep, dogs, and calves. However, whether this organism persists in horses or induces long-term clinical abnormalities is not known. OBJECTIVES: To evaluate whether A. phagocytophilum can persist in horses and to document clinical findings for 3 months after complete recovery from acute disease. ANIMALS: Five clinically normal adult horses that had recovered spontaneously from experimentally induced acute disease caused by a Swedish equine isolate of A. phagocytophilum. METHODS: Horses were monitored for up to 129 days post inoculation (PI) by daily clinical examination and at least alternate day blood sampling for evidence of A. phagocytophilum on polymerase chain reaction (PCR) and blood smears. All horses were euthanized and underwent postmortem examination. RESULTS: All horses were periodically PCR positive after recovery from acute infection. Before day 66 PI 2 horses were persistently PCR negative whereas 3 horses were intermittently PCR positive. Subsequently, 4 of 5 horses were intermittently PCR positive, particularly after stress mimicking interventions. One animal was positive immediately before postmortem examination. Clinical abnormalities related to persistence of anaplasma were not observed. No specific changes were found at postmortem examination, and all sampled tissues from all horses were negative on PCR for A. phagocytophilum. CONCLUSIONS AND CLINICAL IMPORTANCE: Infection with A. phagocytophilum can persist in the horse for at least 129 days. However, the continued presence of the organism is not associated with detectable clinical or pathological abnormalities.

Enilconazole treatment of horses with superficial Aspergillus spp. rhinitis.

BACKGROUND: Mycotic rhinitis is an uncommon disease in horses with a wide range of clinical signs and treatment options based on case studies of a single or only a few individuals. Moreover, while dogs with mycotic rhinitis appear to respond well to treatment with topical enilconazole, this approach has yet to be reported in horses. OBJECTIVES: The aim of this study was to describe the clinical findings, results of diagnostic investigations, treatment, and outcome in horses with mycotic rhinitis treated with topical enilconazole. ANIMALS: Eight horses were included in the study. METHODS: Medical records from 1995 to 2007 from an equine veterinary teaching hospital were reviewed to identify cases with a diagnosis of mycotic rhinitis. RESULTS: Eleven cases were identified, of which 8 were treated with topical enilconazole. Seven of those 8 horses recovered and had normal appearance of the nasal passages on endoscopic examination. Fungal culture, where positive, yielded only Aspergillus spp. Treatment was readily performed and no adverse effects were noted in these horses. In 5 horses, long-term follow-up revealed complete recovery for an average of 5 years. CONCLUSIONS AND CLINICAL IMPORTANCE: Mycotic rhinitis owing to Aspergillus spp. in horses can be effectively and safely treated with topical enilconazole.

Sows intramammarily inoculated with Escherichia coli influence of time of infection, hormone concentrations and leucocyte numbers on development of disease.

The aim of this study was to identify factors that influence the development of disease in sows inoculated with Escherichia coli in the mammary gland. Ten cross-bred primiparous sows were intramammarily inoculated with living E. coli bacteria at different time points before parturition: seven sows within 48 h before parturition and three sows approximately 96 h before parturition. Before and after inoculation, blood samples and mammary gland biopsy specimens were collected and clinical observations were made. All seven sows inoculated close to parturition developed a rectal temperature of >39.5 degrees C during the first 48 h post-partum and two of them also showed other signs of clinical disease. In the sows inoculated 4 days before parturition, the rectal temperature never exceeded 39.5 degrees C during the first 48 h post-partum and none of them showed any other sign of clinical discase. There was a tendency (P < 0.1) that histological signs of mastitis were more frequent in the sows inoculated close to parturition. There were no overall differences between the two groups of sows in plasma concentrations of cortisol, oestradiol-17beta and 15-ketodihydro-PGF2alpha before inoculation. Before inoculation, the number of neutrophils in the blood was overall higher (P < 0.05) in the group of sows that were inoculated close to parturition. In comparison, the number of lymphocytes before inoculation had a tendency (P < 0.1) to be lower in that group. The data suggest that the time of infection of the mammary gland relative to parturition and the number of circulating neutrophils at the time of infection influence the development of chinical coliform mastitis in the sow.

Renal dysplasia in boxers and Finnish harriers.

Puppies from two litters of dogs were found to have severe polyuria and polydipsia. Four of the dogs were investigated by means of clinical examination, haematological and biochemical analysis, and urinalysis. A modified water deprivation response test was also performed in two of the dogs. Renal changes on postmortem examination in three of the dogs were found to be consistent with renal dysplasia. A possible explanation for the finding of hyposthenuria and the extreme polyuria and polydipsia in association with renal dysplasia may be lack of response to antidiuretic hormone owing to anomalous maturation of the renal tubules. Six other puppies from the two litters of dogs did not show any clinical signs of polyuria and polydipsia, although postmortem examination in one of them also revealed renal dysplasia. The clinical features of renal dysplasia may therefore vary greatly between individuals.

Detection of granulocytic Ehrlichia species DNA by PCR in persistently infected dogs.

Three female beagle dogs inoculated with granulocytic Ehrlichia species were monitored for four to six months to determine whether there was evidence that the organisms persisted. The dogs were inoculated intravenously with blood containing an Ehrlichia species closely related to Ehrlichia equi and Ehrlichia phagocytophila, and identical to the human granulocytic ehrlichiosis agent with respect to its 16S rRNA gene sequence. The clinical signs were evaluated, and blood samples were collected for haematology, serum biochemistry and serology. Ehrlichial inclusions in the blood were monitored by microscopy, and ehrlichial DNA was detected by the polymerase chain reaction (PCR). Two of the dogs were injected with prednisolone on days 54 to 56 and days 152 to 154 after infection, and the other was injected with prednisolone on days 95 to 97 after infection. The dogs were euthanased and examined postmortem. Ehrlichial inclusions were demonstrated in the neutrophils and seroconversion occurred shortly after inoculation. Two of the dogs developed acute disease with rectal temperatures above 39.0 degrees C, after which no further clinical signs were observed. The administration of corticosteroids seemed to facilitate the detection of ehrlichial inclusions. Ehrlichial DNA was detected intermittently by PCR in blood samples from two of the dogs throughout the study. Persistent infection was demonstrated up to five-and-a-half months after inoculation.

Early manifestations of granulocytic ehrlichiosis in dogs inoculated experimentally with a Swedish Ehrlichia species isolate.

Seven beagles were inoculated experimentally with a Swedish canine Ehrlichia species isolate to study its pathogenicity. With respect to the 16S rRNA gene sequence, the isolate was identical to the human granulocytic ehrlichiosis (HGE) agent and closely related to both Ehrlichia equi and E phagocytophila. After an incubation period of four to 11 days, the most prominent clinical signs were high fever for two to five days and depression. All the dogs developed profound thrombocytopenia, moderate leukopenia and a strong serological antibody response. Ehrlichial inclusions were detected in blood neutrophils from four to 14 days after inoculation for four to eight days. Ehrlichial DNA could be detected by polymerase chain reaction during the parasitaemic stage and a few days before and after microscopic inclusions were visible. Postmortem, the dogs showed reactive splenic hyperplasia and non-specific mononuclear reactive hepatitis.