Quantitative analysis in spontaneous canine anal sac gland adenomas and carcinomas

Stained cytological specimens from 7 canine anal sac gland adenomas and 11 canine anal sac gland carcinomas were analyzed by computer-assisted nuclear morphometry. In each case, the nuclei of at least 100 neoplastic cells were measured, and the mean nuclear area (MNA), mean nuclear perimeter (MNP), mean nuclear diameter (MND) and nuclear roundness (NR) were calculated. The study aimed to evaluate (1) the possibility of using nuclear cytomorphometry as an auxiliary diagnostic method to differentiate between canine anal sac gland adenomas and adenocarcinomas, and (2) the prognostic value of nuclear morphometry in canine anal sac gland adenocarcinomas. The results indicated that (1) MNA, MNP, MND and NR could be used as effective auxiliary tools for differential diagnosis between canine anal sac gland adenomas and adenocarcinomas, and (2) MNA, MNP and MND are reliable prognostic indicators for canine anal sac gland adenocarcinomas.     

Clinical stage, therapy, and prognosis in canine anal sac gland carcinoma

BACKGROUND: Reports of canine anal sac gland carcinoma (ASGC) describe varied clinical presentations and management and differing responses to therapy. A unifying approach to clinical stage determination and management of this disease has yet to be presented. HYPOTHESIS: An ordinal clinical staging scheme for canine ASGC can be devised on the basis of responses to therapy for a retrospective cohort of affected dogs. ANIMALS: 130 dogs with naturally occurring ASGC. METHODS: A simplified clinical stage system and a management algorithm for canine ASGC were derived from retrospective evaluation of a cohort of 80 dogs; applicability of both was then prospectively evaluated in a cohort of 50 dogs. RESULTS: Retrospective evaluation revealed 4 statistically significant negative prognostic indicators for survival: lack of therapy, presence of distant metastases, presence of lymph node metastases, and primary tumor size. Lymph node extirpation was a statistically significant positive prognostic indicator by bivariate analysis. In both retrospective and prospective analyses, the modified clinical stage scheme revealed a significant association with survival time. CONCLUSIONS AND CLINICAL IMPORTANCE: The clinical staging scheme permits differentiation between groups in terms of prognosis and, therefore, decisions on therapy. This will facilitate application of appropriate therapy and enhanced communication and collaboration in further investigations of ASGC.     

Evaluation of clinical, macroscopic, and histopathologic response to treatment in nonhypoproteinemic dogs with lymphocytic-plasmacytic enteritis

BACKGROUND: Lymphocytic-plasmacytic enteritis (LPE) is a common cause of chronic vomiting and diarrhea in dogs. However, little information is available about endoscopic or histopathologic improvement after therapy in dogs with LPE. HYPOTHESIS: The objective was to study the clinical, endoscopic, and histopathologic evolution of LPE during and after immunosuppressive treatment with prednisone and metronidazole. Most dogs also were treated symptomatically with metoclopramide and cimetidine. ANIMALS: Sixteen dogs with LPE and normal serum protein concentrations diagnosed at the Veterinary Medical Teaching Hospital of the Complutense University of Madrid were monitored during and after drug treatment. The control group consisted of 9 dogs that had no gastrointestinal signs for the preceding 12 months. METHODS: In this prospective clinical treatment trial, clinical, endoscopic, and histopathologic scores were evaluated to describe disease evolution during conventional therapy. Dogs with LPE were monitored for 120 days from the start of treatment. Re-evaluation was performed on post-treatment days 30, 60, 90 (end of treatment), and 120. RESULTS: The average disease activity index observed in our study fell progressively from its initial value, and the decrease between consecutive re-evaluations was statistically significant until day 60 (P = .04). Our results indicate that 75% of the animals revealed improvement of endoscopic gastric lesions (defined as a reduction of the endoscopic score) after treatment, and 75% exhibited improvement of endoscopic duodenal lesions. Statistical analysis of the data revealed significant differences between pre- and post-treatment gastric and duodenal macroscopic endoscopic lesions (P < .05). On the other hand, treatment did not lead to any significant changes in the severity of the gastric and duodenal histopathologic lesions of the affected dogs. CONCLUSIONS AND CLINICAL IMPORTANCE: Treatment of nonhypoproteinemic dogs with LPE led to clinical and endoscopic improvement, but histopathologic lesions were unchanged during therapy.     

Kinetics of Plasma Cell‐Free DNA and Creatine Kinase in a Canine Model of Tissue Injury

Background

Cell‐free DNA (cfDNA) comprises short, double‐stranded circulating DNA sequences released from damaged cells. In people, cfDNA concentrations correlate well with disease severity and tissue damage. No reports are available regarding cfDNA kinetics in dogs.

Objectives/Hypothesis

Cell‐free DNA will have a short biological half‐life and would be able to stratify mild, moderate, and severe tissue injury. Our study aims were to determine the kinetics and biological half‐life of cfDNA and to contrast them with those of creatine kinase (CK).

Animals

Three groups of 10 dogs undergoing open ovariohysterectomy, surgery for cranial cruciate ligament rupture (CCLR), or hemilaminectomy.

Methods

Plasma for cfDNA and CK analysis was collected at admission, at induction of anesthesia, postsurgery (time 0) and at 6, 12, 24, 36, 48, 60, and 72 hours after surgery.

Results

The biological half‐life of plasma cfDNA and CK were 5.64 hours (95% confidence interval [CI 95], 4.36–7.98 hours) and 28.7 hours (CI95, 25.3–33.3 hours), respectively. In the hemilaminectomy group, cfDNA concentrations differed significantly from admission at 6–12 hours after surgery. Creatine kinase activity differed among the surgical groups and reached a peak 6 hours after surgery. In the ovariohysterectomy and CCLR groups, plasma CK activity 72 hours after surgery did not differ from admission activity of the ovariohysterectomy group. In contrast, in the hemilaminectomy group, plasma CK activity after 72 hours did not return to the ovariohysterectomy group admission activity.

Conclusions and Clinical Importance

Plasma CK activity has a longer biological half‐life than previously thought. In contrast to plasma CK activity, cfDNA has a short half‐life and could be a useful marker for peracute severe tissue injury.     

Ultrasound-Guided Cytology of Spleen and Liver: A Prognostic Tool in Canine Cutaneous Mast Cell Tumor

Background: In the clinical staging of cutaneous mast cell tumors (cMCT), the diagnosis of metastasis is controversial based on cytological examination of lymph nodes, spleen, liver, bone marrow, and blood.
Objectives: To define the prognostic role of ultrasound-guided cytology of spleen and liver in cMCT. The results of cytological evaluation were compared in relation with survival time.
Animals: Fifty-two client-owned dogs with a diagnosis of cMCT.
Methods: Selection of cases was based on cytological evaluation of liver and spleen to detect infiltration at distant sites. The Kaplan Meier method was used to compare survival in dogs with and without infiltration of spleen and liver (log-rank test P < .05).
Results: Ten dogs with cMCT had mast cell infiltration of spleen, liver, or both and 4 of these dogs had involvement of the regional lymph nodes. The majority of dogs had 2 or more ultrasonographically abnormal findings simultaneously in spleen and liver. Nine dogs had grade II cMCT, and 1 had grade III cMCT. Dogs with positive evidence of mast cell infiltration to spleen, liver, or both had shorter survival times (34 versus 733 days) compared with dogs negative for mast cell infiltration at distant sites.
Conclusion and Clinical Importance: Dogs with evidence of mast cell infiltration at distant sites have a shorter survival times than dogs without evidence of infiltration at distant sites. This study suggests that cytology of spleen and liver is indicated either for ultrasonographically normal or for ultrasonographically abnormal spleen and liver in dogs with cMCT.     

Pattern-recognition receptor mRNA expression and function in canine monocyte/macrophages and relevance to canine anal furunuclosis

Pattern-recognition receptors (PRRs) are important components of the innate immune system, enabling early detection of infection. Defective PRR function has been implicated in several infectious and immune-mediated diseases of human beings, including Crohn's disease (CD). Anal furunculosis (AF) is an immune-mediated disease which primarily occurs in German shepherd dogs (GSD) and could result from a similar type of PRR dysfunction. The aim of the current study was to investigate canine PRR responses in vitro and to test the hypothesis that these were altered in AF-affected GSD. The pattern-recognition receptors TLR1, TLR2, TLR4, TLR6, TLR9, NOD1 (nucleotide-binding oligomerisation domain) and NOD2 were evaluated in the DH82 canine monocyte/macrophage cell line. These cells were found to express mRNA for all the selected PRRs with TLR2 mRNA the most and TLR5 mRNA the least abundant. A similar pattern of expression was found in canine blood-derived monocyte/macrophages. Stimulation of DH82 cells and blood-derived monocyte/macrophages using specific PRR-ligands, resulted in expression of pro-inflammatory cytokine mRNA. Quantification of TNFalpha mRNA and protein secretion from stimulated cells demonstrated variable responses with lipopolysaccharide (TLR4 ligand) and PAM(3)CSK4 (TLR1/2 ligand) proving to be the most potent and CpG DNA (TLR9 ligand) the least potent. Comparing PRR responses in blood-derived monocyte/macrophages from healthy blood-donor dogs with those from AF-affected GSD showed a deficiency in the latter in response to LD-MDP (NOD2 ligand) at the mRNA level but not at the protein level. It is possible that dysfunctional NOD2 responses by cells of the monocyte/macrophage lineage are involved in the pathogenesis of AF.     

Feline hyperthyroidism: spectrum of clinical presentions and response to carbimazole therapy

OBJECTIVE: To determine the spectrum of clinical presentations of hyperthyroidism in cats and response to carbimazole therapy by analysis of historical, clinical and laboratory data. DESIGN: A prospective clinical study involving client-owned cats presenting to a private veterinary practice in Australia. PROCEDURE: Twenty-five cats diagnosed as hyperthyroid during a 23-month period participated in the study with owner consent. Therapy with carbimazole was instituted and revisits were scheduled 2, 6 and 13 weeks after diagnosis. The cats were physically examined and underwent haematological and serum biochemical testing at each revisit. Owners were also asked to assess clinical signs in their cats in the periods between veterinary examinations. Cats with underlying renal disease were managed by alterations or cessation of carbimazole therapy. RESULTS: A high prevalence of lethargic or inappetent cats without detectable underlying nonthyroidal illness was found. There was also a high prevalence of cats less than 10-years-old and cats in good body condition. Fourteen cats treated with carbimazole and monitored for 13 weeks responded favourably to therapy. Side-effects were minor and uncommon. Cats with underlying renal disease that became apparent during the study, responded well to alterations or cessation of carbimazole therapy. CONCLUSION: The trend towards more subtle clinical presentations of hyperthyroid cats reported previously continued in this study. The findings of the current study do not appear to support the traditional view of hyperthyroid cats as being old, hyperactive, hungry and thin. Carbimazole therapy was found to decrease the prevalence of almost all clinical abnormalities in 14 cats and side-effects were minor and uncommon. This study demonstrates the usefulness of medical management of hyperthyroidism in the cat when radioiodine therapy is not possible due to renal compromise or other factors.     

Canine dilated cardiomyopathy: a retrospective study of signalment, presentation and clinical findings in 369 cases

O bjective : To review the clinical and diagnostic findings and survival of dilated cardiomyopathy from a large population of dogs in England.
M ethods : A retrospective study of the case records of dogs with dilated cardiomyopathy collected between January 1993 and May 2006.
R esults : There were 369 dogs with dilated cardiomyopathy of which all were pure-bred dogs except for four. The most commonly affected breeds were dobermanns and boxers. Over 95 per cent of dogs weighed more than 15 kg and 73 per cent were male. The median duration of signs before referral was three weeks with 65 per cent presenting in stage 3 heart failure. The most common signs were breathlessness (67 per cent) and coughing (64 per cent). The majority of dogs (89 per cent) had an arrhythmia at presentation and 74 per cent of dogs had radiographic signs of pulmonary oedema or pleural effusion. The median survival time was 19 weeks.
C linical S ignificance : Dilated cardiomyopathy occurs primarily in medium to large breed pure-bred dogs, and males are more frequently affected than females. The duration of clinical signs before referral is often short and the survival times are poor. Greater awareness of affected breeds, clinical signs and diagnostic findings may help in early recognition of this disease which often has a short clinical phase.     

Low concentrations of cerebrospinal fluid GABA correlate to a reduced response to phenobarbital therapy in primary canine epilepsy

In this study, we investigated whether pretreatment cerebrospinal fluid (CSF) neurotransmitter concentrations of gamma-aminobutyric acid (GABA) and glutamate (GLU) were correlated with response to phenobarbital treatment in dogs with primary epilepsy. Eleven untreated dogs, 6 males and 5 females, with a median age of onset of seizures of 3 years (range: 0.5-5 years) were selected for therapy based on progressive or serious seizure patterns. The median interval between the first observed seizure and start of phenobarbital therapy was 485 days (range: 101-1,765 days). All dogs were purebred, with the exception of I male dog. Oral phenobarbital was started at 2.5 mg/kg every 12 hours. Trough serum phenobarbital concentrations were measured at 15, 45, 90, 180, 360, 540, and 720 days after the start of treatment. There was no difference in the mean trough serum concentration or in the mean number of seizures recorded between each time period of phenobarbital measurement over the 2-year evaluation. No correlation was found between CSF GLU, GABA, or GLU: GABA ratio and the total number of seizures recorded before or after initiation of phenobarbital therapy. Lower CSF GABA concentration, however, was correlated with a lower seizure frequency difference (the total number of seizures before phenobarbital therapy minus the total number of seizures after phenobarbital therapy for an identical time period of evaluation) and lower percentage reduction in seizures: ([total number of seizures before phenobarbital therapy minus the total number of seizures after phenobarbital therapy] divided by the total number of seizures before phenobarbital therapy) x 100. There was no correlation between CSF GLU and the seizure frequency difference and percentage reduction in seizures. A negative correlation between the CSF GLU:GABA ratio and seizure frequency difference was found. Thus, dogs with an initial lower CSF GABA concentration before phenobarbital therapy did not respond as well as did dogs with a higher CSF GABA concentration.     

Evaluation of a Portable Meter to Measure Ketonemia and Comparison with Ketonuria for the Diagnosis of Canine Diabetic Ketoacidosis

Background: The diagnosis of canine diabetic ketoacidosis (DKA) usually is based on measurement of urinary acetoacetate (ketonuria). In humans, this test is less sensitive and specific than blood 3-β-hydroxybutyrate (ketonemia) evaluation.
Hypothesis: Ketonemia measurement using a portable meter is more accurate than ketonuria determination with a dipstick to diagnose canine DKA.
Animals: Seventy-two client-owned diabetic dogs with ketonemia, ketonuria, or both.
Methods: Prospective observational study. Based on blood bicarbonate concentration and anion gap, dogs were divided into 2 groups: patients with DKA ( n = 25); patients with diabetic ketosis ( n = 47). Sensitivity, specificity, and positive and negative likelihood ratio (LR) at different cut-off points were determined for both ketonemia and ketonuria. Receiver operating characteristic (ROC) analysis was used to assess the accuracy of each diagnostic test to diagnose DKA.
Results: With regard to ketonemia, cut-off values of 2.3 and 4.3 mmol/L revealed 100% sensitivity and 100% specificity, respectively, whereas cut-off values of 2.8 and 3.5 mmol/L showed a −LR of 0.05 and a + LR of 13.16, respectively. With regard to ketonuria, a cut-off value of 1+ revealed 92% sensitivity, 40% specificity, and −LR of 0.20, whereas a cut-off value of 3+ revealed 44% sensitivity, 94% specificity, and +LR of 6.89. The areas under the ROC curves for the ketonemia and ketonuria tests were significantly different (0.97 and 0.81, respectively, P = .003).
Conclusions and Clinical Importance: Measurement of ketonemia is accurate and more effective than measurement of ketonuria to diagnose canine DKA.     

Canine Subcutaneous Zygomycosis Caused by Conidiobolus sp.: A Case Report and Review of Conidiobolus Infections in other Species

Résumé— Un chien, jeune adulte, a été présenté pour une masse sous-cutanée évoluant depuis un mois sur la partie droite du thorax et une masse ulcérée sous-cutanée sur la cuisse droite. L'examen histo-pathologique a révélé une inflammation pyogranulomateuse du derme et du tissu sous-cutané avec de larges hyphes fongiques. De nouvelles lésions sont apparues. Un traitement à base d'itraconazole pendant 2 semaines a été inefficace. L'euthanasie a été pratiquée 10 semaines après la première consultation. La culture fongique faite à partir de prélèvements chirurgicaux a permis d'isoler un Conidiobolus sp. Des infections àConidiobolus sp. ont été rapportées chez l'homme, le cheval, le mouton et le lama. Histologiquement, les hyphes sont difficiles à différencier de ceux d'autres zygomycètes ou de Pythium insidiosum. La culture est nécessaire pour une identification définitive. Une discussion sur les aspects comparés des infections cutanées àConidiobolus sp. est développée. [Hiller, A., Kunkle, G. A., Ginn, P. E., Padhye, A. A. Canine subcutaneous zygomycosis caused by Conidiobolus sp.: a case report and review of Conidibolus infections in other species. (Zygomycose sous-cutanée àConidiobolus sp. chez un chien: cas clinique et revue des infections àConidiobolus dans les autres espèces). Resumen— Un perro adulto joven presentaba una masa subcutánea en la región torácica derecha y otra ulcerada en la parte proximal de la extremidad posterior derecha, ambas de un mes de duración. El exámen histopatológico mostró una inflamación piogranulomatosa de la dermis y del tejido subcutáneo con grandes hifas füngicas. A pesar de que la enfermedad cutánea presentó un curso oscilante, aparecieron nuevas lesiones. Un tratamiento de dos semanas con itraconazol no alteró el curso de la enfermedad. El animal fue eutanasiado a las 10 semanas de la primera visita. Se ais lóConidiobolus sp. a partir de las muestras quirúrjicas. Las infecciones por Conidiobolus sp. estan descritas en la especie humana, la equina, la ovina y en las llamas. Las hifas no se diferencian fácilmente a nivel histológico de otros cigomicetales o de Pythium insidiosum y es necesaria la identificatión mediante un cultivo fúngico para establecer un diagnóstico definitive Se discuten las caracteristicas de la infección cutánea por Conidiobolus sp. y se describen las caracteristicas de la pitiosis cutánea. [Hillier, A., Kunkle, G. A., Ginn, P. E. and Padhye, A. A. Canine subcutaneous zygomycosis caused by Conidiobolus sp.: a case report and review of Conidibolus infections in other species (Cigomcosis subcutánea canina causada por Conidibolus sp.: un caso clinico y revisién de las infecciones por Conidibolus sp. en otras especies). Zusammenfassung— Ein junger ausgewachsener Hund wurde vorgestellt, der seit einem Monat an der rechten Thoraxwand ein subkutanes Gebilde und proximal an der rechten Hintergliedmaße eine ulzer-ierende, subkutane Masse aufwies. Bei der histopathologischen Untersuchung wurde eine pyogranulomatöse Entzündung der Dermis und Subkutis mit breiten Pilzhyphen entdeckt. Trotz des “Auf und Ab” der Hautkrankheit erschionon laufend neue Hautveränderungen. Eine zweiwöchige Behandlung mit Itrakonazol veränderte den Verlauf der Krankheit nicht. 10 Wochen nach der Erstvorstellung des Patienten wurde en euthenesiert. In der Kultur von chrirugischen Proben wurde Conidiobolus sp. isoliert. Conidiobolus sp.-Infektionen sind bei Menschen, Pferden, Schafen und Lamas beschrieben. Die Hyphen können histologisch schlecht von denen anderen Zygomyzeten oder von Pythium insidiosum unterschieden werden. Für die endgültige Diagnose ist eine Identifizierung mittels Pilzkultur notwendig. Die Erscheinungsbilder von kutanen Conidiobolus sp.-Infektionen werdon diskutiert und mit denen kutaner Phytiosis verglichen. [Canine subcutaneous zygomycosis caused by Conidiobolus sp.: a case report and reviews of Conidiobolus infections in other species (Subkutane Zygomykose des Hundes durch Conidiobolus sp.: Fallbericht und übersicht über Conidioblus-Infektionen bie anderen Tierarten). Abstract— A young adult dog was presented with a 1-month history of a subcutaneous mass on the right thoracic wall and an ulcerative, subcutaneous mass of the proximal right hindlimb. Pyogranulomatous inflammation of the dermis and subcutis with broad fungal hyphae were found on histopathologic examination. Despite some waxing and waning of the skin disease, new lesions continued to appear. A 2-week course of itraconazole did not alter the course of the disease. Euthanasia was performed 10 weeks after initial presentation. Conidiobolus sp. was isolated in culture from surgical specimens. Conidiobolus sp. infection has been reported in humans, horses, sheep and llamas. The hyphae cannot be easily distinguished histologically from those of some other zygomycetes or Pythium insidiosum and fungal culture identification of the organism is required for definitive diagnosis. A discussion of the features of cutaneous Conidiobolus sp. infection is given and comparative features of cutaneous pythiosis are presented.     

Methods for diagnosis of canine leishmaniasis and immune response to infection

Canine leishmaniasis (CanL) caused by Leishmania infantum (syn. L. chagasi, in Latin America), which is transmitted by the bite of phlebotomine sand flies, is endemic and affects millions of dogs in Europe, Asia, North Africa and South America. It is an emergent disease in North America. Early detection and treatment of infected animals may be critical in controlling the spread of the disease and is an essential part of human zoonotic visceral leishmaniasis control. The laboratory diagnosis of CanL still poses a challenge, despite progress made in the development of several direct and indirect methods. An effective diagnosis test, apart of being able to confirm a clinical suspicion in a single patient as well as to detect infection in asymptomatic dogs, should have high sensitivity, specificity and reproducibility; it must be simple, easy to perform, non-expensive, feasible in regional laboratories or adaptable for field conditions. Ideally, it should detect all Leishmania-infected dogs, preferentially using non-invasive collection of biological samples. In this paper we review the advantages and shortcomings of the available procedures for CanL diagnosis in the different phases, e.g. pre-patent and patent period of the infection and methods to determine the related immune response.     

Treatment of advanced canine anal sac adenocarcinoma with hypofractionated radiation therapy: 77 cases (1999–2013)

Currently no standard of care exists for advanced, inoperable or metastatic anal sac adenocarcinoma (ASAC). The objective of this retrospective study was to assess the role of hypofractionated radiation therapy (RT) in 77 dogs with measurable ASAC. A total of 38% of dogs experienced a partial response to RT. For dogs presenting with clinical signs related to the tumour, improvement or resolution of signs was noted in 63%. For dogs presenting with hypercalcemia of malignancy, resolution was noted in 31% with RT alone and an additional 46% with radiation, prednisone, and/or bisphosphonates. Median overall survival was 329 days (range: 252–448 days). Median progression free survival was 289 days (range: 224–469). There was no difference in survival based on radiation protocol, use of chemotherapy, previous surgery or advanced stage. Radiation toxicities were mild and infrequent. Hypofractionated RT is well tolerated and is applicable in the treatment of advanced primary, locoregional or metastatic ASAC.