Clinical techniques in small animal molecular oncology

The diagnosis, staging, and treatment of tumors in veterinary as well as human oncology have traditionally incorporated elements of anatomic extent of neoplasia through various clinical and pathologic methods. These clinicopathological methods have been the basis for the development of the tumor, node, and metastasis and grading systems, which have translated into clinically significant advances over the last 20 to 30 years. Unfortunately, there continues to be significant limitations to this system when prognostication and therapeutic decisions need to be made specific to a patient. For example, completely resected and cleanly staged phenotypically identical grade II mast cell tumors in dogs can have opposing clinical outcomes. In addition, dogs or cats with identical stage and grade lymphoma can have significantly divergent responses to the same multi-agent chemotherapy protocol. Numerous nonanatomic neoplastic molecular prognostic factors have been recently identified and have the potential to improve on the presently available tumor, node, and metastasis- and grading-based systems. To date in human oncology, single nonanatomic factors have only occasionally translated into efficient and independent prognostic factors, which speaks to the heterogeneity of cancer. Therefore, the use of panels of factors have been encouraged that will allow for the development of a molecular prognostic index, which can then be used in concert with presently available systems. This review will summarize how to best utilize presently available tumor, node, and metastasis- and grading-based systems, and incorporate newly available molecular prognostic factors.     

Evaluation of the effects of ischemic injury and ureteral obstruction on delayed graft function in cats after renal autotransplantation

OBJECTIVE: To determine the relative importance of ischemic injury to delayed graft function (DGF) in cats. STUDY DESIGN: Experimental study. ANIMALS: Six intact female cats. METHODS: Cats had renal autograft transplantation without ureteral transection and reimplantation and a contralateral nephrectomy. Serum creatinine and blood urea nitrogen (BUN) concentrations were measured regularly and abdominal ultrasound was performed before surgery, the day after surgery and twice weekly thereafter. Ultrasound-guided renal biopsy was performed on day 7. Cats were euthanatized on day 21. Histology of the autograft, ureter, bladder, vascular anastomoses sites, and contralateral kidney were performed. Observations were compared with those from an historic group of research cats that had extravesicular ureteroneocystostomy and contralateral nephrectomy. RESULTS: Five cats completed the study. Serum creatinine and BUN concentrations increased after surgery, peaking at 3.2+/-0.8 and 77.6+/-15.9 mg/dL, respectively, 1-2 days after surgery. Serum creatinine concentration returned to the reference interval by 6 days after surgery. BUN gradually decreased in all cats but did not return to the reference interval by study end. Serum creatinine and BUN concentrations were consistently lower but not significantly so (P=.29 and .56, respectively) compared with the historic ureteroneocystostomy group. No ultrasonographic abnormalities or renal biopsy histologic abnormalities were observed. At necropsy, 1 autograft had generalized interstitial fibrosis. CONCLUSION: Harvesting the renal graft and the ischemia before revascularization causes impaired renal function after engraftment. CLINICAL RELEVANCE: The process of harvesting and reimplanting the renal graft can contribute to DGF in cats, independent of ureteral obstruction.     

头骨形态指标鉴别水貂来源有效性的评估

人工驯养野生动物是获取野生动物产品的重要手段,但是非法盗猎的野生动物产品以人工饲养的名义进入市场,就会对野生动物的保护形成挑战。因此,准确鉴别市场上的动物产品是来自养殖场还是野外,是保护野外种群,维持养殖业正常秩序的关键。本研究以北美水貂(Neovison vison)为例,基于颧骨内侧空间(ZAS)、颧宽(ZAA和ZBA)、颅基长(BSL)、下颌长(MDL)、眶后收缩(POC)和最大面部宽度(GFB)等量度,建立了9个头骨形态计量学指标:POC/GFB、POC/BSL、POC/MDL、ZAS SQRT/BSL、ZA SSQRT/MDL、ZBA×BSL/2ZAS、ZBA×MDL/2ZAS、ZBB×BSL/2ZAS和ZBB×MDL/2ZAS。野生组(n=32,9雄23雌)和饲养组(n=45,35雄10雌)的比较表明,除POC/GFB外,所有指标对于在两组样品之间均有显著差异,整体判别正确率为73.2%—85.5%。在性别已知的情况下,雄性的整体正确率为82.4%—93.8%,雌性为41.7%—85.7%。为此,应针对不同性别、不同来源的动物建立相应的参考数据,利用似然比进行鉴别。本研究的结果不仅可用于北美水貂的鉴别,也为其他小型食肉动物的鉴别提供经验。     

Utility of plasma N-terminal pro-brain natriuretic peptide (NT-proBNP) to distinguish between congestive heart failure and non-cardiac causes of acute dyspnea in cats

BackgroundCirculating plasma N-terminal pro-brain natriuretic peptide (NT-proBNP) concentration facilitates emergency diagnosis of congestive heart failure (CHF) in people. Its utility to discriminate between dyspneic cats with CHF vs. primary respiratory disease requires further assessment. Our objectives were to determine if NT-proBNP (1) differentiates dyspneic cats with CHF vs. primary respiratory disease; (2) increases with renal insufficiency; (3) correlates with left atrial dimension, radiographic cardiomegaly, and estimated left ventricular filling pressure (E/E_a).MethodsNT-proBNP was measured in 167 dyspneic cats (66 primary respiratory disease, 101 CHF) to evaluate (1) relationship with clinical parameters; (2) ability to distinguish CHF from primary respiratory disease; (3) optimal cut-off values using receiver operating characteristic (ROC) curve analysis.ResultsNT-proBNP (1) was higher (median and inter-quartile [25th–75th] percentile) in CHF (754 pmol/L; 437, 1035 pmol/L) vs. primary respiratory disease (76.5 pmol/L; 24, 180 pmol/L) cohorts (P < 0.001); (2) positively correlated in CHF cats with increased inter-ventricular septal end-diastolic thickness (ρ = 0.266; P = 0.007) and LV free wall thickness (ρ = 0.218; P = 0.027), but not with radiographic heart size, left atrial size, left ventricular dimensions, E/E_a ratio, BUN, creatinine, or thyroxine; (3) distinguished dyspneic CHF cats from primary respiratory disease at 265 pmol/L cut-off value with 90.2% sensitivity, 87.9% specificity, 92% positive predictive value, and 85.3% negative predictive value (area under ROC curve, 0.94).ConclusionsNT-proBNP accurately discriminated CHF from respiratory disease causes of dyspnea.     

Canine oral melanoma

Melanoma is the most common oral malignancy in the dog. Oral and/or mucosal melanoma has been routinely considered an extremely malignant tumor with a high degree of local invasiveness and high metastatic propensity. Primary tumor size has been found to be extremely prognostic. The World Health Organization staging scheme for dogs with oral melanoma is based on size, with stage I = <2-cm-diameter tumor, stage II = 2- to <4-cm-diameter tumor, stage III = > or = 4cm tumor and/or lymph node metastasis, and stage IV = distant metastasis. Median survival times for dogs with oral melanoma treated with surgery are approximately 17 to 18, 5 to 6, and 3 months with stage I, II, and III disease, respectively. Significant negative prognostic factors include stage, size, evidence of metastasis, and a variety of histologic criteria. Standardized treatments such as surgery, coarse-fractionation radiation therapy, and chemotherapy have afforded minimal to modest stage-dependent clinical benefits and death is usually due to systemic metastasis. Numerous immunotherapeutic strategies have been employed to date with limited clinical efficacy; however, the use of xenogeneic DNA vaccines may represent a leap forward in clinical efficacy. Oral melanoma is a spontaneous syngeneic cancer occurring in outbred, immunocompetent dogs and appears to be a more clinically faithful therapeutic model for human melanoma; further use of canine melanoma as a therapeutic model for human melanoma is strongly encouraged. In addition, the development of an expanded but clinically relevant staging system incorporating the aforementioned prognostic factors is also strongly encouraged.     

Bacterial community analysis of purulent material from liver abscesses of crossbred cattle and Holstein steers fed finishing diets with or without tylosin

Liver abscesses in feedlot cattle are polymicrobial infections. Culture-based studies have identified Fusobacterium necrophorum as the primary causative agent, but a number of other bacterial species are frequently isolated. The incidence of liver abscesses is highly variable and is affected by a number of factors, including cattle type. Holstein steers raised for beef production have a higher incidence than crossbred feedlot cattle. Tylosin is the commonly used antimicrobial feed additive to reduce the incidence of liver abscesses. The objective of this study was to utilize 16S ribosomal RNA amplicon sequence analyses to analyze the bacterial community composition of purulent material of liver abscesses of crossbred cattle (n = 24) and Holstein steers (n = 24), each fed finishing diet with or without tylosin. DNA was extracted and the V3 and V4 regions of the 16S rRNA gene were amplified, sequenced, and analyzed. The minimum, mean, and maximum sequence reads per sample were 996, 177,070, and 877,770, respectively, across all the liver abscess samples. Sequence analyses identified 5 phyla, 14 families, 98 genera, and 102 amplicon sequence variants (ASV) in the 4 treatment groups. The dominant phyla identified were Fusobacteria (52% of total reads) and Proteobacteria (33%). Of the top 25 genera identified, 17 genera were Gram negative and 8 were Gram positive. The top 3 genera, which accounted for 75% of the total reads, in the order of abundance, were Fusobacterium, Pseudomonas, and Bacteroides. The relative abundance, expressed as percent of total reads, of phyla, family, and genera did not differ (P > 0.05) between the 4 treatment groups. Generic richness and evenness, determined by Shannon–Weiner and Simpson’s diversity indices, respectively, did not differ between the groups. The UniFrac distance matrices data revealed no clustering of the ASV indicating variance between the samples within each treatment group. Co-occurrence network analysis at the genus level indicated a strong association of Fusobacterium with 15 other genera, and not all of them have been previously isolated from liver abscesses. In conclusion, the culture-independent method identified the bacterial composition of liver abscesses as predominantly Gram negative and Fusobacterium as the dominant genus, followed by Pseudomonas. The bacterial community composition did not differ between crossbred and Holstein steers fed finishing diets with or without tylosin.