Ultrasonographic evaluation of normal canine iliopsoas muscle

The aim of this study was to describe the normal ultrasonographic appearance of the iliopsoas muscle and related landmarks. Hind limbs of five dog cadavers with no history of lameness were evaluated. The origin and mid-body of the psoas major and the common insertion of the iliacus and psoas major on the lesser trochanter of the femur were identified and evaluated. New methylene blue was injected under ultrasonographic guidance at the three sites. Dissection was performed to confirm placement of the dye. The L3 origin, mid-body, and insertion of the muscle were identified in all dogs and were consistent in appearance and compatible with the general appearance of muscle and tendons. The L2 origin was consistently difficult to image. The same ultrasound technique was subsequently applied to four healthy dogs, and consistent images of the iliopsoas muscle and associated landmarks were obtained. In this study, the major structures of the iliopsoas could be identified and in all dogs had a similar appearance. Ultrasound is an important tool for the diagnosis of musculotendinous injury and may be useful for identification of ilipsoas injury as a cause of lameness in the dog.     

同时检测犬瘟热病毒和犬细小病毒双重PCR方法的建立

为建立可以同时检测犬瘟热病毒(CDV)和犬细小病毒(CPV)的双重PCR方法,本研究根据GenBank登录的CDV N蛋白序列和CPV NS基因保守序列,设计合成2对特异性引物。通过优化反应条件,对CDV阳性病毒株反转录后的cDNA模板和CPV的DNA模板进行双重PCR扩增,同时得到2条与试验设计相符的669 bp(CDV)和392 bp(CPV)特异性条带,建立了同时检测CDV和CPV的双重PCR方法。实验结果表明:在同一PCR反应体系中可以同时检测这2种病毒,而对犬腺病毒Ⅰ型、犬腺病毒Ⅱ型、狂犬病毒检测均为阴性;CDV和CPV的最低检出限分别为101.8TCID50和101.4TCID50。采用该方法对在黑龙江省不同地区所采集的30份犬病料样品进行检测,CDV阳性率为30%;CPV阳性率为23.33%,表明建立的PCR方法可以用于临床诊断。     

Retrospective comparison of the efficacy of fluconazole or itraconazole for the treatment of systemic blastomycosis in dogs

Background: Itraconazole is recommended for treatment of blastomycosis in dogs. Some evidence suggests that fluconazole might be less hepatotoxic than itraconazole. Objectives: To compare (1) incidence of clinical remission and death; (2) treatment duration; (3) total drug cost; (4) incidence of relapse; and (5) incidence of increased ALT activities in dogs with blastomycosis treated with fluconazole or itraconazole. Animals: One hundred and forty‐four dogs with systemic blastomycosis treated with itraconazole or fluconazole from 1998 to 2008. Methods: Retrospective case review. Information obtained included signalment, body weight, clinical signs, drug regimen, treatment duration, time to clinical remission, and laboratory results. Results: Neither treatment efficacy between fluconazole (75% remission) and itraconazole (90% remission) nor relapse rate (18% for itraconazole, 22% for fluconazole) was significantly different (P= .13, .75, respectively). Treatment duration was significantly longer for fluconazole (median 183 days) than for itraconazole (138 days; P= .001). Costs for fluconazole (median $1,223) were significantly less than for itraconazole ($3,717; P < .001). Incidence of increased ALT activities was not significantly different between groups (17% [3/18] for fluconazole, 26% [6/23] for itraconazole; P= .71). Conclusions: Fluconazole is associated with survival to clinical remission in 75% of dogs with blastomycosis. Although dogs receiving fluconazole were treated longer, drug costs were one‐third those of itraconazole. Hepatotoxicosis, as estimated by increases in serum ALT activity, can be observed with similar incidence for both drugs.     

Evaluation of a multidrug chemotherapy protocol with mitoxantrone based maintenance (CHOP-MA) for the treatment of canine lymphoma

The purpose of this study was to evaluate the efficacy of adding mitoxantrone to a cyclophosphamide, doxorubicin, vincristine, l -asparaginase and prednisone containing protocol. Sixty-five dogs with multicentric lymphoma were evaluated for overall remission and survival times. Remission and survival time versus stage, substage, pretreatment hypercalcaemia and pretreatment steroid administration were also evaluated. Overall median remission for dogs with multicentric lymphoma was 302 days and overall median survival was 622 days. Of the dogs with multicentric lymphoma, 23 (35%) received all scheduled mitoxantrone doses. Only median survival versus substage was found to be significant (substage a median survival was 679 days and substage b median survival was 302 days, P = 0.025). Increasing the total combined dose of doxorubicin and mitoxantrone may improve remission times when compared with historical controls, and further studies are needed to determine how best to utilize mitoxantrone in multidrug chemotherapy protocols for canine multicentric lymphoma.     

Isolation and identification of a canine coronavirus strain from giant pandas (Ailuropoda melanoleuca)

Two giant pandas (Ailuropoda melanoleuca) died of unknown causes in a Chinese zoo. The clinical disease profile suggested that the pandas may have suffered a viral infection. Therefore, a series of detection including virus isolation, electron microscopy, cytobiological assay, serum neutralization and RT-PCR were used to identify the virus. It was determined that the isolated virus was a canine coronavirus (CCV), on the basis of coronavirus, neutralization by canine anti-CCV serum, and 84.3% to 100% amino acid sequence similarity with CCV. The results suggest that the affected pandas had been infected with CCV.     

表达犬瘟热病毒H基因重组新城疫病毒的构建及其免疫原性研究

为研制安全、有效的新型犬瘟热疫苗,本研究利用新城疫病毒(NDV) LaSota弱毒疫苗株反向遗传操作系统,构建出表达犬瘟热病毒(CDV)弱毒疫苗株Rockborn-20/8血凝素(H)蛋白的重组病毒rLa-CDV-H,并对其生物学特性进行鉴定,评估其作为犬瘟热活载体疫苗的安全性和有效性.通过免疫荧光和western blot试验证明了CDV H蛋白的正确表达;重组病毒株保持了LaSota亲本株的低致病性和高滴度鸡胚生长特性;重组病毒rLa-CDV-H接种12周龄比格犬后,可以诱导显著的CDV中和抗体反应.本研究表明重组病毒rLa-CDV-H具有作为犬瘟热重组病毒活载体疫苗的潜力.     

Evaluation of ifosfamide salvage therapy for metastatic canine osteosarcoma

A retrospective study was performed to assess toxicity and response rate of ifosfamide salvage treatment for dogs diagnosed with metastatic osteosarcoma (OSA). Dogs diagnosed with OSA and previously treated with standard chemotherapy were included in the study. Nineteen dogs met the inclusion criteria, and 17 dogs were evaluable for response. Ifosfamide doses ranged from 375 to 425 mg m^?2 (median dose 375 mg m^?2), with a median of two doses administered per dog (range 1–7 doses). The overall response to ifosfamide was 11.8% [complete response (CR) = 1/17, partial response (PR) = 1/17, stable disease (SD) = 2/17, progressive disease (PD) = 13/17]. Two dogs were hospitalized due to ifosfamide toxicosis. The median survival duration from the first dose of ifosfamide to death was 95 days. Ifosfamide was well tolerated, but minor anti‐tumour activity was observed.     

Double‐blinded cross‐over study on the efficacy of pentoxifylline for canine atopy

The pathogenesis of canine atopy has not been established completely. Recent studies have shown that tumour necrosis factor alpha and Interleukin‐6 play a role in allergic reactions in humans and mice. Pentoxifylline (PTX) suppresses synthesis of these cytokines and may be a useful therapy for modulating the symptoms of canine atopy. The objectives of this study were to investigate the effects of PTX (10mg kg^?1 twice daily for 4 weeks) on clinical signs (erythema and pruritus) and intradermal skin test reactivity in atopic dogs (n = 10). The study was a double‐blinded, placebo controlled, crossover clinical trial with a washout period of 2 weeks between treatments. Clinical signs were evaluated and scored by the investigator and owners. During PTX treatment, scores of pruritus and erythema decreased significantly. PTX did not affect intradermal skin test reactivity to house dust mite at 15 min (allergen of reference for this study).     

Computed tomographic-dacryocystography (CT-DCG) of the normal canine nasolacrimal drainage system with three-dimensional reconstruction

The aims of the study were (1) to quantify the influence of selected imaging parameters on the image quality (slice thickness, mAs, and beam orientation) defining optimal conditions for scan protocols and (2) to evaluate the benefits of the 3D reconstruction techniques for visualization of NDS structures in dogs. CT-DCG was performed bilaterally in 32 heads of dogs. CT transverse images were obtained using a combination of two slice thickness values (0.8 mm and 2 mm) and two mAs values (50 mAs and 300 mAs). Two beam projection orientations were also tested: transverse plane (perpendicular to the hard palate) and oblique to the hard palate. Three-dimensional images were obtained using Volume Rendering (VR). Transverse beam projection proved to be superior for the assessment of the inferior and superior lacrimal canaliculi and lacrimal sac. In this study, there was no statistical difference regarding mAs values (50 mAs and 300 mAs) and slice thickness values (0.8 mm and 2 mm). Three-dimensional images were helpful for the assessment of topographic relationship between nasolacrimal structures and cranial landmarks.     

Use of canine small intestinal submucosa allograft for treating perineal hernias in two dogs

Here, we describe two dogs in which canine small intestinal submucosa (SIS) was implanted as a biomaterial scaffold during perineal herniorrhaphy. Both dogs had developed severe muscle weakness, unilaterally herniated rectal protrusions, and heart problems with potential anesthetic risks. Areas affected by the perineal hernia (PH) located between the internal obturator and external anal sphincter muscles were reconstructed with naïve canine SIS sheets. In 12 months, post-operative complications such as wound infections, sciatic paralysis, rectal prolapse, or recurrence of the hernia were not observed. Symptoms of defecatory tenesmus also improved. Neither case showed any signs of rejection or specific immune responses as determined by complete and differential cell counts. Our findings demonstrate that canine SIS can be used as a biomaterial scaffold for PH repair in dogs.     

Oral melphalan for the treatment of relapsed canine lymphoma

Oral melphalan has been included in multi‐agent rescue protocols for canine lymphoma but its activity as a single‐agent for this purpose has not been established. Inexpensive cost, ease of administration and tolerability make oral melphalan an attractive candidate for single‐agent rescue therapy of canine lymphoma. Retrospective evaluation of 19 cases of relapsed canine lymphoma treated with oral melphalan was performed. Melphalan was primarily administered (n = 16) via a high dose protocol (HDM ) with a median dosage of 19.4 mg m^?2. Fifteen dogs (78.9%) were treated concurrently with corticosteroids. Response evaluation was possible for all dogs with a calculated overall clinical benefit (partial response [PR ] + stable disease [SD ]) of 31.6% (PR 3/19; SD 3/19). Times to progression following melphalan (TTP ‐M) were 14, 24 and 34 days for responders and 20, 28 and 103 days for dogs experiencing SD . Twelve of 17 dogs evaluable for toxicity experienced an adverse event (AE ) with only 3 dogs experiencing a grade III or higher AE . Haematologic toxicity was common (11/17) while gastrointestinal toxicity was rare (1/17). Although treatment resulted in limited clinical benefit and non‐durable responses, oral melphalan was well‐tolerated and may be a reasonable rescue option in cases where minimal effective agents remain.     

Efficacy and safety of suberoylanilide hydroxamic acid (Vorinostat) in the treatment of canine corneal fibrosis

Objective Study aims were to evaluate the safety and efficacy of the Food and Drug Administration‐approved drug Vorinostat [suberoylanilide hydroxamic acid (SAHA)] in the treatment of canine corneal fibrosis using an in vitro model. Methods Healthy donor canine corneas were collected and used to generate primary canine corneal fibroblasts (CCFs) by growing cultures in minimal essential medium supplemented with 10% fetal bovine serum. Canine corneal myofibroblasts, used as a model for corneal fibrosis, were produced by growing CCF cultures in serum‐free medium containing transforming growth factor β1 (1 ng/mL). Trypan blue exclusion assays were used to determine the optimal SAHA dose for this in vitro model. Four hour after culturing with TGFβ1, CCF cultures were treated with 0.06% SAHA for 5 min (group 1) and for 24 h (group 2), representing single and multiple dose treatment regimes, respectively. Cultures were then further incubated in the presence of TGFβ1 (1 ng/μL) under serum‐free conditions until they reached 70% confluence. Trypan blue exclusion, immunocytochemistry, and TUNEL assays were used to evaluate the cytotoxicity of SAHA. Real‐time PCR, western blot analysis, and immunocytochemistry were used to determine the efficacy of SAHA to inhibit canine corneal myofibroblast formation. Results Topical SAHA application in both treatment groups successfully decreased α‐smooth muscle actin expression when compared to the TGFβ1 only treatment group (P < 0.05). Tested SAHA did not affect CCF phenotype or cellular viability and did not cause significant cell death. Conclusions Suberoylanilide hydroxamic acid safely and effectively inhibits TGFβ1‐induced CCFs transformation to myofibroblast in vitro.     

Mycobacterial nodular granulomas affecting the subcutis and skin of dogs (canine leproid granuloma syndrome)

Objective To obtain a better understanding of a disease affecting dogs in which nodular mycobacterial granulomas are present in the subcutis or skin. Design Retrospective survey Procedure A diagnosis of an unidentified mycobacterial infection was made in 45 dogs following detection of acid-fast bacilli surrounded by granulomatous or pyogranulomatous inflammation in tissue sections. Eight cases were identified from the records of our diagnostic laboratory. In all cases culture for mycobacteria was unsuccessful. Another 37 cases were identified by practitioners and veterinary pathologists in response to a survey mailed to veterinary clinics throughout Australia. Data from these 45 cases were entered into a data base. The data set was incomplete, as some respondents did not answer all questions. Results Over 90% of affected dogs had short coats and about half were Boxers or Boxer-cross dogs. A weak seasonal trend for the development of primary lesions in autumn and winter was identified. Cases were recorded from New South Wales (35 dogs), Western Australia (4), Queensland (4), Tasmania (1) and New Zealand (1). The subcutis and skin of the ears and head (including the ears) were involved in 64% and 85% of cases, respectively, for which lesion site was recorded. Culture was attempted but was unsuccessful in 19 cases, including 11 cases where material was submitted to our laboratory and/or a Mycobacteria Reference Laboratory. Lymph node enlargement, internal organ involvement or constitutional signs were not a feature of the syndrome, and most lesions did not worry the dogs. Although many practitioners recorded a favourable response to therapy with doxycycline (response rate 57%) or amoxycillin-clavulanate (63%), spontaneous resolution of infection was thought to have occurred in six of seven dogs (86%) not given systemic antimicrobials, and three dogs where antimicrobials had failed previously. A minority of cases failed to respond to antimicrobial therapy and continued to have chronic lesions. There was no discernible trend for dogs of a particular age or sex to be affected. Conclusion This syndrome is caused by saprophytic mycobacteria of limited pathogenicity that give rise to lesions restricted principally but not exclusively to the subcutis and skin of body extremities. Fastidious growth requirements have prevented their isolation on synthetic media used for culture of mycobacteria. Organisms presumably enter the subcutis following a breach in integrity of the epidermal barrier and produce self-limiting disease in immunocompetent dogs. Lesions tend to resolve spontaneously. The possibility of a public health threat from affected dogs is highly unlikely.     

An in vitro evaluation of storage media for the preservation of canine packed red blood cells

The effect of four different red blood cell storage media on in vitro parameters of stored canine red blood cells was studied. The storage media included citrate-phosphate-dextrose-adenine (CPDA-1), two additive solutions, and an additive solution modified by the addition of plasma. Biochemical and hematologic parameters, including red cell adenosine triphosphate (ATP); 2,3-diphosphoglycerate (2,3-DPG); pH; percent hemolysis; and supernatant sodium, potassium, and glucose were assessed immediately following preparation of the red cell concentrate and after 35 and 42 days of storage at 4 degrees C. All parameters changed significantly (p < 0.05) during storage. Significant differences due to effect of the storage media were also seen at each time period. After 35 days and 42 days of storage, CPDA-1 maintained the highest pH, potassium, and sodium values, and had the lowest 2,3-DPG, ATP (p=0.052), and glucose values. No differences were seen in hemolysis after 35 days of storage. No additional benefit was noted from the addition of plasma to the additive solution. The additive solutions compared favorably with CPDA-1.     

A randomized controlled trial of misoprostol monotherapy for canine atopic dermatitis: effects on dermal cellularity and cutaneous tumour necrosis factor-alpha

Abstract In this blinded randomized placebo-controlled trial, 20 dogs with atopic dermatitis (AD) were given placebo (8 dogs) or misoprostol (12 dogs) at 5 µg kg⁻¹, orally, three times daily for 3 weeks. Administration of the active drug, but not of placebo, led to a significant decrease in lesional and pruritus scores. The median reduction from baseline of both scores was ≈30%. Misoprostol therapy did not lead to decreases of dermal cell counts or skin tumour necrosis factor (TNF)α mRNA copy numbers that were significantly different from those of placebo. Skin TNFα protein production, assessed using indirect immunofluorescence, decreased or remained unchanged in dogs receiving misoprostol. In contrast, post treatment TNFα fluorescence scores were higher in all but two dogs given placebo. The changes from baseline of TNFα fluorescence scores did not correlate significantly with those of lesional or pruritus indices. These observations confirm the modest efficacy of misoprostol for treatment of canine AD and suggest that its mild anti-allergic effects are not associated with either inhibition of inflammatory cell emigration or TNFα production.     

MAGNETIC RESONANCE IMAGING OF CEREBRAL CORTICAL NECROSIS (POLIOENCEPHALOMALACIA) IN A DOG

A 3-year-old neutered female mixed breed dog was examined because of severe, generalized seizure activity, tetraparesis, and encephalopathic signs. Cerebrospinal fluid (CSF) evaluation was unremarkable except for a mild increase in protein. Serum and CSF titers for infectious diseases were negative. Magnetic resonance (MR) imaging examination of the brain was performed and lesions were found within the cerebral gray matter of the temporal and parietal lobes. The lesions had increased signal intensity on T1, T2, and proton density-weighted images. There was mild inhomogeneous enhancement following intravenous contrast medium administration. Neurologic status improved and the seizures were well controlled, but the dog never regained normal mentation and euthanasia was performed 10 weeks after initial evaluation. At necropsy, severe cerebral cortical necrosis was found in the regions corresponding to the lesions seen on MR imaging examination. Large numbers of fat-containing macrophages (gitter cells) were found within these areas, and are thought to be responsible for the characteristic hyperintensity seen on the MR images.     

犬细小病毒荧光定量PCR检测方法的建立与评价

通过序列比对和Blast分析,选定犬细小病毒(Canine parvovirus,CPV)VP2蛋白保守区基因为检测的目的基因,引物采用Primer Premier 5.0软件设计。利用灵敏度较高的TaqMan探针法建立CPV核酸检测方法。通过对标准品的扩增、测序及对标准扩增曲线的绘制,建立CPV核酸检测方法。同时对建立的检测方法进行了检测特异性、灵敏度和重复性分析。将阳性对照标准品进行10倍梯度稀释后可检测到102拷贝/μL样品,表明该检测体系具有较高的检测灵敏度。通过分析表明,本检测方法在用空白对照及类似的猪细小病毒、猪圆环病毒作为扩增对照时,没有发现非特异性产物的产生,表明该体系对于CPV的检测是特异的。通过6次批间重复检测,体系的变异系数小于3%,表明该检测体系具有良好的重复性。