Toxicity of halogenated oxyquinolines in dogs. A clinical study. 3. Intoxication experiments

LANNEK, BIRGITTA and PAUL LINDBERG: Toxicity of halogenated oxyquinolines in dogs. A clinical study. III. Intoxication experiments. Acta vet. scand. 1974, 15, 398–418. — Oxyquinoline drugs, which are normally well tolerated by dogs, will cause disease when a dog’s resistance is occasionally lowered. In a series of experiments in dogs we tried to reproduce the disease pattern of spontaneous cases of poisoning. Most of these experiments, using even high single or repeated oral doses, failed. It was observed, more or less by chance, that the intestinal absorption of ¹²⁵I-labelled vioform was greatly increased when the dog was not fasted. The consumption of fat, but not of protein or carbohydrates, was found to be the responsible factor. When the oxyquinoline drug was given in a fat emulsion, approx. 1/3 of the dogs fell ill. When fouled fish was also added, the dose necessary to produce disease was lowered to the range used in vioform therapy. We believe that phenolic substances, which may be produced from bacterial degradation of proteins in intestinal disorders, compete with oxyquinolines in metabolic and elimination processes.vioform; oxyquinolines; dogs; convulsions; heart injury; liver injury; poisoning; diarrhoea; phenolic substances.     

Toxicity of halogenated oxyquinolines to dogs. The effect of feeding fouled herring

Acute clinical toxicity of halogenated oxyquinolines in dogs has been reported (Schantz & Wikström 1965, Hangartner 1965, Müller 1967). The clinical picture was comprehensively studied (Lannek 1972 b). The dose range in the clinical material was found to vary from about 7 to 250 mg per kg body weight (Lannek 1972 a).     

C-reactive protein as an indicator of inflammatory responses to experimentally induced cystitis in dogs

The aim of this study was to demonstrate and assess C-reactive protein (CRP) changes in dogs with induced bacterial cystitis with or without antibiotics. We also evaluated availability of CRP levels to serve as an indicator for monitoring or diagnosing bacterial cystitis. Serial CRP concentrations in dogs with induced bacterial cystitis were higher than those of controls (p < 0.001). CRP concentrations peaked on day 7 and gradually decreased thereafter. In the treatment group, CRP concentrations decreased after medication compared to the untreated group (p = 0.032). CRP levels had a linear correlation with urine white blood cell counts among all groups (r = 0.837, p < 0.001, n = 140). Compared to the negative urine culture group, dogs with positive urine culture results had higher CRP concentrations (median 43.8 mg/L vs. 5.9 mg/L; p < 0.001). Area under the receiver operating characteristic curve was 0.955; when cut-off value was 12.2 mg/L, CRP measurements were found to have a sensitivity of 92.3% and specificity of 86.4%. This result indicates that rapid increases of CRP occurred after inducing bacterial cystitis and CRP may be a useful indicator for monitoring or diagnosing canine bacterial cystitis together with sediment urinalysis and urine bacterial culture.     

Dogs are more permissive than cats or guinea pigs to experimental infection with a human isolate of Bartonella rochalimae

Bartonella rochalimae was first isolated from the blood of a human who traveled to Peru and was exposed to multiple insect bites. Foxes and dogs are likely natural reservoirs for this bacterium. We report the results of experimental inoculation of two dogs, five cats and six guinea pigs with the only human isolate of this new Bartonella species. Both dogs became bacteremic for 5–7 weeks, with a peak of 10³–10⁴ colony forming units (CFU)/mL blood. Three cats had low bacteremia levels (< 200 CFU/mL) of 6–8 weeks’ duration. One cat that remained seronegative had two bacterial colonies isolated at a single culture time point. A fifth cat never became bacteremic, but seroconverted. None of the guinea pigs became bacteremic, but five seroconverted. These results suggest that dogs could be a reservoir of this strain of B. rochalimae, in contrast to cats and guinea pigs.     

Serum Phospholipase A2 in Canine Acute Pancreatitis

During 3 years 28 cases of acute pancreatitis were diagnosed in dogs. In 26 of these dogs, the disease was fatal. The most frequent symptoms were vomiting, anorexia and lethargy. Two thirds showed tenderness upon abdominal palpation. Ascites was found in 3 cases. Of the blood, parameters, serum amylase level was elevated in 86 % and lipase in 89 % of the cases. Sixteen dogs were uremic and half of the dogs were hyperglycemic. Two thirds of the dogs had leukocytosis. Using stepwise multiple regression the best blood parameters explaining acute pancreatitis were leukocytes together with lipase and glucose.In an attempt to find a more specific serum test for dogs to diagnose acute pancreatitis serum phospholipase A2 (PLA₂) activity was measured. In sixteen out of the 28 dogs with acute pancreatitis, serum PLA₂ activity was increased. The ascites fluids were rich in PLA2. Serum PLA₂ is more often increased in the severe necrotizing pancreatitis (80 %) than in the milder forms of acute pancreatitis (44 %). All dogs with increased serum PLA₂ had also increased serum amylase and lipase activities. The dogs with an increased serum PLA₂ and dogs with ascites had fat necrosis in the vicinity of the pancreas. Experimental pancreatitis was induced in 4 dogs by injecting Na-taurocholate and trypsin into the pancreas. In these cases, very high PLA₂ activities in the serum and ascites fluids were detected, but none seemed to be present in the urine samples.Key words: dog, acute pancreatitis, phospholipase A2     

Evaluation of urine specific gravity as a predictor of hypotension during anaesthesia in healthy dogs premedicated with dexmedetomidine

ObjectiveTo investigate the relationship between urine specific gravity (USG) and the risk of arterial hypotension during general anaesthesia (GA) in healthy dogs premedicated with dexmedetomidine and methadone.Study designProspective clinical cohort study.AnimalsA total of 75 healthy client-owned dogs undergoing GA for elective tibial plateau levelling osteotomy.MethodsAfter placing an intravenous catheter, dogs were premedicated with dexmedetomidine (5 μg kg^–1) and methadone (0.3 mg kg^–1) intravenously. After induction of GA with alfaxalone to effect, the bladder was expressed and USG measured. An arterial catheter was placed, and residual blood was used to measure packed cell volume (PCV) and total protein (TP). GA was maintained with isoflurane vaporised in oxygen and a femoral and sciatic nerve block were performed. Arterial blood pressure < 60 mmHg was defined as hypotension and recorded by the anaesthetist. Treatment for hypotension was performed in a stepwise manner following a flow chart. Frequency of hypotension, treatment and response to treatment were recorded. Logistic regression modelling was used to assess the association between USG, TP and PCV and incidence of perioperative hypotension; p < 0.05.ResultsData from 14 dogs were excluded. Of the 61 dogs, 16 (26%) were hypotensive during GA, 15 dogs needed treatment of which 12 were responsive to a decrease in inhalant vaporiser setting. The logistic regression model was not statistically significant (p = 0.8). There was no significant association between USG (p = 0.6), TP (p = 0.4), PCV (p = 0.8) and arterial hypotension during GA.Conclusions and clinical relevanceIn healthy dogs premedicated with dexmedetomidine and methadone and maintained under GA with isoflurane and a femoral and sciatic nerve block, there was no relationship between the specific gravity of urine collected after premedication and intraoperative arterial hypotension.     

Effects of morphine and fentanyl constant rate infusion on urine output in healthy and traumatized dogs

ObjectiveTo determine whether healthy and traumatized dogs receiving a constant rate infusion (CRI) of either morphine or fentanyl have decreased urine production.Study designProspective randomized controlled study.Animal populationEighteen privately owned previously healthy dogs that had undergone trauma were included. Twenty-three privately owned healthy dogs were used as the controls.MethodsTraumatized dogs were randomized into one of two groups. Group Tmorphine received a CRI of morphine (0.12 mg kg⁻¹ hour⁻¹) and group Tfentanyl received a CRI of fentanyl (3 μg kg⁻¹ hour⁻¹) both administered in lactated Ringer’s solution (LRS) at a rate of 60 mL kg⁻¹ day⁻¹. Control healthy dogs were randomized into one of three groups. The LRS control group (CLRS) (n = 8) received LRS at a rate of 60 mL kg⁻¹ day⁻¹. Group Cmorphine (n = 8) and group Cfentanyl (n = 7) received the same infusions as Tmorphine and Tfentanyl, respectively. Collected data were identical for all groups and consisted of measuring total fluid administered, urine output, and urine specific gravity (USG) for a 24-hour period. An analysis of variance (anova) was used for statistical analysis and a p < 0.05 was considered statistically significant.ResultsUrine output was significantly decreased (p < 0.05) in all groups compared with the LRS control group. The end mean USG was significantly lower (p = 0.003) in the LRS control group compared with all other groups.ConclusionsThere was a decrease in urine output with a CRI of morphine or fentanyl in both healthy and traumatized dogs.Clinical relevanceDecreased urine output caused by an opioid effect might lead to improper assessments of renal function and urine production.     

An Open‐label Phase 1 Dose‐escalation Clinical Trial of a Single Intravenous Administration of Gemcitabine in Dogs with Advanced Solid Tumors

Background

A broad range of gemcitabine dosages have been used in dogs.

Hypothesis/Objectives

To determine maximally tolerated dose (MTD), dose‐limiting toxicity (DLT), and preliminary antitumor activity of intravenous administration of gemcitabine in dogs with advanced solid tumors.

Animals

Twenty‐two client‐owned dogs.

Methods

Dogs with advanced cancer were prospectively enrolled in an open‐label Phase 1 study of gemcitabine. Gemcitabine was administered as a 30‐minute intravenous bolus starting at 800 mg/m², using escalation of 50 mg/m² increments with 3 dogs per dose level. MTD was established based on the number of dogs experiencing DLT assessed after 1 cycle. Treatment continued until disease progression or unacceptable toxicosis. Additional dogs were enrolled at MTD to better characterize tolerability, and to assess the extent and duration of gemcitabine excretion.

Results

Twenty‐two dogs were treated at 4 dose levels, ranging from 800 to 950 mg/m². Neutropenia was identified as DLT. MTD was 900 mg/m². DLT consisting of grade 4 febrile neutropenia was observed at 950 mg/m² in 2 dogs. There were no nonhematologic DLTs. Twenty dogs received multiple doses, and none had evidence of severe toxicosis from any of their subsequent treatments. At 900 mg/m², 2 complete and 5 partial responses were observed in dogs with measurable tumors. The amount of gemcitabine excreted in urine decreased over time, and was undetectable after the first 24 hours.

Conclusions and Clinical Importance

The recommended dose of gemcitabine for future Phase 2 studies is weekly 900 mg/m². In chemotherapy‐naïve dogs with advanced solid tumor this dose level merits further evaluation.     

Hypercoagulability in Dogs with Blastomycosis

Background

Blastomycosis is a potentially fatal fungal disease that most commonly affects humans and dogs. The organism causes systemic inflammation and has a predilection for the lungs. The inflammation might lead to a hypercoagulable state with microemboli in the pulmonary circulation which could contribute to inadequate oxygen exchange in infected dogs.

Hypothesis/Objectives

Dogs with blastomycosis will be hypercoagulable compared with healthy case‐matched controls.

Animals

Client‐owned dogs with a diagnosis of blastomycosis (n = 23) and healthy case‐matched controls (n = 23).

Methods

Prospective case‐controlled study of client‐owned dogs presented to a veterinary teaching hospital with clinical signs compatible with blastomycosis. Complete blood counts, fibrinogen, PT, aPTT, thromboelastometry (TE), thrombin antithrombin complexes (TAT), and thrombin generation were evaluated.

Results

Cases had a leukocytosis compared with controls [mean (SD) 16.6 (7.6) × 10³/μL versus 8.2 (1.8) × 10³/μL, P < .001], hyperfibrinogenemia [median 784 mg/dL, range 329–1,443 versus median 178 mg/dL, range 82–257, P < .001], and increased TAT concentrations [mean (SD) 9.0 (5.7) μg/L versus 2.0 (2.8) μg/L, P < .001]. As compared to controls, cases were also hypercoagulable as evaluated by thromboelastometry and had increased in vitro thrombin generation on calibrated automated thrombography.

Conclusions and Clinical Importance

Hypercoagulability occurs in dogs with systemic blastomycosis. Additional studies are needed to explore a possible contribution of thrombogenicity to the clinical manifestations of systemic blastomycosis.     

Evaluation of Iron Deficiency Using Reticulocyte Indices in Dogs Enrolled in a Blood Donor Program

Background

People donating blood more than twice annually are at risk of developing iron deficiency. Little is known about the iron status of dogs enrolled in blood donor programs.

Hypothesis

Dogs donating blood ≥6 times annually will show evidence of iron deficiency based on their reticulocyte indices.

Animals

Thirteen dogs enrolled in a blood donor program donating ≥6 times over the preceding 12 months and 20 healthy nondonor control dogs.

Methods

Prospective observational study. Mature red blood cell (RBC) indices, reticulocyte indices, serum iron, serum ferritin, and total iron‐binding capacity (TIBC) were compared between groups.

Results

Packed cell volume (median 47%, range 40–52%, P < .01), hematocrit (median 46.4%, range 40.3–52.5%, P < .01), and reticulocyte count (median 16,000/μL, range 9,000–38,000/μL, P < .01) were significantly lower in the blood donor dogs. No statistically significant differences were noted in the mature RBC indices between groups. Both reticulocyte mean corpuscular volume (median 88.8 fL, range 83.4–95.5 fL, P = .03) and reticulocyte hemoglobin content (median 24.6 pg, range 23.1–26.6 pg, P < .01) were significantly lower in the blood donor group. Serum iron and ferritin were similar between groups; however, TIBC was significantly higher in the control group (median 403 μg/dL, range 225–493 μg/dL, P = .02).

Conclusions

The findings in dogs donating ≥6 times annually suggest the presence of iron‐deficient erythropoiesis in this population.     

Incidence,Severity and Prognosis Associated with Hyponatremia in Dogs and Cats

Background

Hyponatremia is a common electrolyte abnormality in human patients and is associated with substantial morbidity and death. The incidence and importance of hyponatremia in dogs and cats has not been determined.

Hypothesis/Objectives

To describe the incidence of and prognosis associated with hyponatremia in dogs and cats at a university teaching hospital.

Animals

Of 16,691 dogs and 4,211 cats with measured blood or serum sodium concentration.

Methods

Retrospective study. Medical records of animals with a blood or serum sodium concentration measured during a 60‐month period were reviewed to determine the severity of hyponatremia and its associated fatality rate. Cases with moderate (11–15 mmol/L below the reference range) or severe hyponatremia (≥16 mmol/L below the reference range) were further reviewed.

Results

Of 4,254 dogs (25.5%) and 2,081 cats (49.4%) were diagnosed with hyponatremia. Case fatality rates of dogs and cats with hyponatremia were 13.7% and 11.9%, respectively, compared to 4.4% and 4.5% with a normal blood or serum sodium concentration (P < 0.0001). The magnitude of hyponatremia was linearly associated with a higher case fatality rate (P < 0.0001). Hyponatremia was associated with a lower case fatality rate than hypernatremia in the same population. Among the animals with moderate or severe hyponatremia, 92.1% of dogs and 90.6% of cats presented with community‐acquired hyponatremia, and 7.9% of dogs and 9.4% of cats developed hospital‐acquired hyponatremia.

Conclusions and clinical importance

Hyponatremia was found commonly in this population and was associated with increased case fatality rate. Presence and severity of hyponatremia might be useful as a prognostic indicator.     

Multidisciplinary approach in the diagnosis of acute leptospirosis in dogs naturally infected by Leptospira interrogans serogroup Icterohaemorrhagiae: A prospective study

Leptospirosis, a zoonotic disease with worldwide distribution, is caused by spirochetes of the genus Leptospira. In dogs, this disease is frequently misdiagnosed. Few studies have attempted to associate the detection of Leptospira spp. infection with clinicopathological and renal histopathological findings using a multidisciplinary approach. The present study isolated and characterized Leptospira spp. obtained from naturally infected dogs and described relevant clinical and histopathological findings. Blood and urine were collected from 57 dogs with clinical symptomatology suggestive of leptospirosis; 38 cases were confirmed by PCR in urine or by culture or microscopic agglutination testing (titers ≥800). A total of 12 strains of pathogenic Leptospira were isolated from the studied dogs (seven in blood, four in urine and one in both blood and urine samples). All isolates were characterized as Leptospira interrogans serogroup Icterohaemorrhagiae. Of the confirmed cases, almost one-third of the animals had been vaccinated. Our analysis of laboratory testing revealed that azotemia and proteinuria were statistically significant predictors of infection. The main histopathological findings seen in kidney tissues were necrosis, degeneration, tubular regeneration, mononuclear inflammatory infiltrate and congestion. A multidisciplinary approach involving clinicopathological and histopathological characterization of renal involvement can aid in the identification of acute leptospirosis infection.     

Incidence,Severity and Prognosis Associated with Hypernatremia in Dogs and Cats

Background

Hypernatremia has been associated with substantial morbidity and death in human patients. The incidence and importance of hypernatremia in dogs and cats has not been determined.

Hypothesis/Objectives

To describe the incidence of and prognosis associated with hypernatremia in dogs and cats at a university teaching hospital.

Animals

A total of 16,691 dogs and 4,211 cats with measured blood or serum sodium concentration.

Methods

Retrospective study. Medical records of animals with a blood or serum sodium concentration measured during a 60‐month period were reviewed to determine the severity of hypernatremia and its associated case fatality rate. Cases with moderate (11–15 mmol/L above the reference range) or severe hypernatremia (≥16 mmol/L above the reference range) were further reviewed.

Results

A total of 957 dogs (5.7%) and 338 cats (8.0%) were diagnosed with hypernatremia. Case fatality rates of dogs and cats with hypernatremia was 20.6 and 28.1%, respectively compared to 4.4 and 4.5% with a normal blood or serum sodium concentration (P < .0001). The magnitude of hypernatremia was linearly associated with a higher case fatality rate (P < .0001). Hypernatremia was associated with a higher case fatality rate than hyponatremia. Among the animals with moderate or severe hypernatremia, 50% of dogs and 38.5% of cats presented with community‐acquired hypernatremia, and 50% of dogs and 61.5% of cats developed hospital‐acquired hypernatremia.

Conclusions and clinical importance

Hypernatremia was found infrequently in this population but was associated with increased case fatality rates in dogs and cats. Presence and severity of hypernatremia might be useful as a prognostic indicator.     

Comparison between Urine Protein: Creatinine Ratios of Samples Obtained from Dogs in Home and Hospital Settings

Background

The urine protein:creatinine ratio (UPC) is used to quantify urine protein excretion and guide recommendations for monitoring and treatment of proteinuria.

Hypothesis/Objectives

Home urine samples will have lower UPCs than hospital samples. The objectives were to compare UPCs of samples collected in each setting and to determine whether environment of sample collection might affect staging, monitoring or treatment recommendations.

Animals

Twenty‐four client‐owned dogs.

Methods

Prospective, nonmasked study. Clients collected a urine sample from their dog at home and a second sample was collected at the hospital. Dogs receiving corticosteroids or angiotensin‐converting enzyme inhibitors were excluded, as were those with urine samples of inadequate volume, no protein on dipstick analysis, or active urine sediment. Samples were refrigerated after collection, dipstick and sediment evaluations were completed and each sample was frozen at −80°C within 12 hours. UPCs were performed on frozen samples within 2 months.

Results

From 81 paired samples, 57 were excluded. Of the remaining 24, 12/24 (50%) had higher hospital sample UPCs, 9/24 (38%) had identical UPCs, and 3/24 (12%) had lower hospital UPCs. The UPCs of hospital samples were higher than home samples for the total population (P = .005) and the subset with UPC > 0.5 (P = .001).

Conclusions

Setting and related circumstances of urine collection in dogs is associated with UPC differences; results are usually higher in hospital than in home samples. This difference has the potential to affect clinical interpretation.     

Flea species infesting dogs in Florida and Bartonella spp. prevalence rates

Several Bartonella spp. associated with fleas can induce a variety of clinical syndromes in both dogs and humans. However, few studies have investigated the prevalence of Bartonella in the blood of dogs and their fleas. The objectives of this study were to determine the genera of fleas infesting shelter dogs in Florida, the prevalence of Bartonella spp. within the fleas, and the prevalence of Bartonella spp. within the blood of healthy dogs from which the fleas were collected. Fleas, serum, and EDTA-anti-coagulated whole blood were collected from 80 healthy dogs, and total DNA was extracted for PCR amplification of Bartonella spp. The genera of fleas infesting 43 of the dogs were determined phenotypically. PCR amplicons from blood and flea pools were sequenced to confirm the Bartonella species. Amplicons for which sequencing revealed homology to Bartonella vinsonii subsp. berkhoffii (Bvb) underwent specific genotyping by targeting the 16S–23S intergenic spacer region.A total of 220 fleas were collected from 80 dogs and pooled by genus (43 dogs) and flea species. Bartonella spp. DNA was amplified from 14 of 80 dog blood samples (17.5%) and from 9 of 80 pooled fleas (11.3%). B. vinsonii subsp. berkhoffii DNA was amplified from nine dogs and five of the flea pools. Bartonella rochalimae (Br) DNA was amplified from six dogs and two flea pools. One of 14 dogs was co-infected with Bvb and Br. The dog was infested with Pulex spp. fleas containing Br DNA and a single Ctenocephalides felis flea. Of the Bvb bacteremic dogs, five and four were infected with genotypes II and I, respectively. Of the Bvb PCR positive flea pools, three were Bvb genotype II and two were Bvb genotype I.Amplification of Bvb DNA from Pulex spp. collected from domestic dogs, suggests that Pulex fleas may be a vector for dogs and a source for zoonotic transfer of this pathogen from dogs to people. The findings of this study provide evidence to support the hypothesis that flea-infested dogs may be a reservoir host for Bvb and Br and that ectoparasite control is an important component of shelter intake protocols.     

Detection and characterization of Leptospira spp. in dogs diagnosed with kidney and/or liver disease in Selangor,Malaysia

Leptospirosis is a serious bacterial disease that affects both humans and animals. A wide range of symptoms have been described in humans; the disease in dogs is commonly associated with kidney and/or liver disease. In Malaysia, information about the common serovars infecting dogs is limited. Therefore, we investigated the occurrences of leptospirosis in 124 pet dogs diagnosed with kidney and/or liver disease. Blood, urine, abdominal effusion, and/or kidney and liver were collected from the dogs. Based on microscopic agglutination testing, 53 of 124 (42.7%) dogs were seropositive for leptospiral exposure. Sera were frequently positive to serovars Bataviae (n = 12), Javanica (n = 10), and Icterohaemorrhagiae (n = 10). Direct detection using PCR showed that 42 of 124 (33.9%) of the whole blood and 36 of 113 (31.9%) urine samples were positive for pathogenic Leptospira spp. By PCR, 2 of 23 (9.1%) kidney and 2 of 23 (9.1%) liver were positive for pathogenic Leptospira spp. Abdominal effusion from 4 dogs were PCR-positive for pathogenic Leptospira spp. The species detected were L. interrogans, L. borgpetersenii, L. kirschneri, and L. kmetyi by partial 16S rRNA sequencing. We further identified and characterized 11 Leptospira spp. isolates from 8 dogs as serovars Bataviae, Javanica, and Australis. The mortality rate of the Leptospira-infected dogs was high (18 of 53; 34%).     

Cohort Study of the Success of Controlled Weight Loss Programs for Obese Dogs

Background

Most weight loss studies in obese dogs assess rate and percentage of weight loss in the first 2–3 months, rather than the likelihood of successfully reaching target weight.

Objective

To determine outcome of controlled weight loss programs for obese dogs, and to determine the factors associated with successful completion.

Animals

143 obese dogs undergoing a controlled weight loss program.

Methods

This was a cohort study of obese dogs attending a referral weight management clinic. Dogs were studied during their period of weight loss, and cases classified according to outcome as “completed” (reached target weight), “euthanized” (was euthanized before reaching target weight), or “stopped prematurely” (program stopped early for other reasons). Factors associated with successful completion were assessed using simple and multiple logistic regression.

Results

87/143 dogs (61%) completed their weight loss program, 11 [8%] died or were euthanized, and the remaining 45 [32%] stopped prematurely. Reasons for dogs stopping prematurely included inability to contact owner, refusal to comply with weight management advice, or development of another illness. Successful weight loss was positively associated with a faster rate (P < .001), a longer duration (P < .001), and feeding a dried weight management diet (P = .010), but negatively associated with starting body fat (P < .001), and use of dirlotapide (P = .0046).

Conclusions and Clinical Relevance

Just over half of all obese dogs on a controlled weight loss program reach their target weight. Future studies should better clarify reasons for success in individual cases, and also the role of factors such as activity and behavioral modification.     

Detection and characterization of Clostridium perfringens in the feces of healthy and diarrheic dogs

Clostridium perfringens has been implicated as a cause of diarrhea in dogs. The objectives of this study were to compare 2 culture methods and to evaluate a multiplex polymerase chain reaction (PCR) assay to detect C. perfringens toxin genes alpha (α), beta (β ), beta 2 (β2), epsilon (ɛ), iota (ι), and C. perfringens enterotoxin (cpe) from canine isolates. Fecal samples were collected from clinically normal non-diarrheic (ND) dogs, (n = 105) and diarrheic dogs (DD, n = 54). Clostridium perfringens was isolated by directly inoculating stool onto 5% sheep blood agar (SBA) and enrichment in brain-heart infusion (BHI) broth, followed by inoculation onto SBA. Isolates were tested by multiplex PCR for the presence of α, β, β2, ɛ, ι, and cpe genes. C. perfringens was isolated from 84% of ND samples using direct culture and from 87.6% with enrichment (P = 0.79). In the DD group, corresponding isolation rates were 90.7% and 93.8% (P = 0.65). All isolates possessed the α toxin gene. Beta (β), β2, ɛ, ι, and cpe toxin genes were identified in 4.5%, 1.1%, 3.4%, 1.1%, and 14.8% of ND isolates, respectively. In the DD group, β and β2 were identified in 5%, ɛ and ι were not identified, and the cpe gene was identified in 16.9% of isolates. Enrichment with BHI broth did not significantly increase the yield of C. perfringens, but it did increase the time and cost of the procedure. C. perfringens toxin genes were present in equal proportions in both the ND and DD groups (P ≤ 0.15 to 0.6). Within the parameters of this study, culture of C. perfringens and PCR for toxin genes is of limited diagnostic usefulness due to its high prevalence in normal dogs and the lack of apparent difference in the distribution of toxin genes between normal and diarrheic dogs.     

Persistent Ehrlichia ewingii Infection in Dogs after Natural Tick Infestation

Background

Ehrlichia ewingii, which causes disease in dogs and people, is the most common Ehrlichia spp. infecting dogs in the United States, but little is known about how long E. ewingii infection persists in dogs.

Hypothesis/Objectives

To evaluate the persistence of natural infection with E. ewingii in dogs.

Animals

Four Class A Beagles; no previous exposure to ticks or tick‐borne infectious agents.

Methods

Dogs were exposed to ticks by weekly walks through tick habitat in north central Oklahoma; dogs positive for infection with Ehrlichia spp. by sequence‐confirmed PCR and peptide‐specific serology were evaluated for 733 days (D). Whole blood was collected once weekly for PCR, and serum was collected once monthly for detection of antibodies to Ehrlichia canis (peptide p16), Ehrlichia chaffeensis (indirect fluorescence antibody [IFA] and variable‐length PCR target [VLPT]), and E. ewingii (peptide p28).

Results

All dogs (4/4) became infected with Ehrlichia spp. as evidenced by seroconversion on IFA to E. chaffeensis (4/4); PCR detection of E. ewingii (4/4) and E. chaffeensis (2/4) DNA using both nested and real‐time assays; and presence of specific antibodies to E. ewingii (4/4) and E. chaffeensis (2/4). Infection with E. chaffeensis was not detected after D55. Intermittent E. ewingii rickettsemia persisted in 3 of 4 dogs for as long as 733 days.

Conclusions and Clinical Importance

Our data demonstrate that dogs infected with E. ewingii from tick feeding are capable of maintaining infection with this pathogen long‐term, and may serve as a reservoir host for the maintenance of E. ewingii in nature.     

Antimicrobial Susceptibility Patterns in Urinary Tract Infections in Dogs (2010–2013)

Background

Urinary tract infections (UTIs) are common in dogs. The responsible bacterial populations have evolved with increasing resistance to many antimicrobials.

Objective

To characterize the antimicrobial susceptibility patterns of canine urinary tract isolates over a 51‐month period.

Animals

One thousand six hundred and thirty‐six bacterial isolates from 1,028 dogs.

Methods

Aerobic bacterial isolate growth and susceptibility data from urine cultures of dogs were identified, retrospectively. Medical records were reviewed to obtain signalment, comorbidities, and antimicrobial use in the previous 30 days. The UTIs were further categorized as uncomplicated, complicated, or pyelonephritis.

Results

Common bacterial isolates identified were Escherichia coli (52.5%), Staphylococcus spp. (13.6%), and Enterococcus spp. (13.3%). In vitro susceptibility among all isolates varied for commonly prescribed antimicrobials (amoxicillin [59%], amoxicillin/clavulanic acid [76%], cephalexin [66%], enrofloxacin [74%] and trimethoprim‐sulfamethoxazole [86%]). For all antimicrobials tested (except aminoglycosides), in vitro susceptibility was higher in uncomplicated versus complicated infections (P < .05). Uncomplicated infection isolate susceptibility rates remained ≤90% for PO administered antimicrobials. Administration of amoxicillin, doxycycline, and enrofloxacin, but not amoxicillin/clavulanic acid in the previous 30 days was associated with resistance to that antimicrobial. Multidrug resistant isolates of E. coli and Staphylococcus spp. were more common in dogs with complicated than uncomplicated UTIs (36% versus 21%, P < .0001).

Conclusions and Clinical Importance

In vitro susceptibility was highly variable and no PO administered antimicrobial had >90% efficacy among isolates tested. Multidrug resistance was frequent among isolates tested suggesting that routine culture and susceptibility testing is indicated. Previously prescribed antimicrobials may affect empirical choices made pending susceptibility testing.