Protein-losing enteropathy in dogs is associated with decreased fecal proteolytic activity

Background: Measurement of proteolytic activity in feces is a traditional method for the diagnosis of exocrine pancreatic insufficiency (EPI). A drawback of this method is the occurrence of falsely low results that may lead to a false‐positive diagnosis of EPI. We hypothesized that intestinal loss of serum proteinase inhibitors in protein‐losing enteropathy (PLE) may inhibit fecal proteolytic activity and be a potential source of false low results. Objective: The objective of this study was to determine the effect of PLE on fecal proteolytic activity in dogs. Methods: Fecal proteolytic activity was measured using a radial diffusion casein digestion assay in 12 samples from 4 clinically healthy control dogs and 30 samples from 16 dogs with PLE. Gastrointestinal protein loss was assessed using an ELISA to determine fecal canine α₁‐proteinase inhibitor concentration. The relationship between the concentration of canine α₁‐proteinase inhibitor in the feces and the diameter cleared in the casein digestion assay was determined. The mean clearing diameter was compared between control dogs and dogs with PLE. Results: A significant negative correlation was observed between fecal canine α₁‐proteinase inhibitor concentration and casein clearing diameter (P < .001, Pearson r=—.6317, r 2 =.3999). Mean clearing diameter was significantly lower in dogs with PLE than in control dogs (12.63 vs 16.83 mm, P < .001, two‐tailed Student's t‐test). Conclusion: Increased fecal loss of α₁‐proteinase inhibitor in dogs with PLE is associated with a significant decrease in fecal proteolytic activity and may result in a false positive diagnosis of EPI.     

NBT-PABA test to assess efficiency and kinetics of substituted proteolytic enzyme action in pancreatic duct ligated minipigs

The NBT-PABA test is an established method for diagnosis of pancreatic exocrine insufficiency. In the present study the NBT-PABA test was used to test and compare the efficacy of two multienzyme preparations (product A and B) differing in galenic preparation in minipigs in which pancreatic exocrine insufficiency (PEI) was induced by pancreatic duct ligation. Without enzyme substitution no distinct increase in PABA was found in blood after oral administration of NBT-PABA. Administration of both enzyme preparations led to a clear dose dependent rise in PABA-concentrations in blood. Interestingly, the two preparations showed different time curves of serum PABA concentration, indicating differences in the kinetic of proteolytic enzyme action. It is concluded that the NBT-PABA test can be a very useful test for indirectly evaluating proteolytic enzyme efficacy in vivo, and also gives information about the kinetics of enzyme action, not only the end-result of enzyme action (like digestibility trials which were used traditionally). A single test is performed in a few hours and there is no need for fistulated animals.     

Comparison of radioimmunoassay and chemiluminescent assay methods to estimate canine blood cortisol concentrations

Background Non‐radioactive assay methods are widely used in commercial laboratories to measure canine blood cortisol concentrations, despite a paucity of published validity data of these tests compared with the traditional ‘gold standard’ radioimmunoassay. Objectives To compare a commercial chemiluminescence assay with radioimmunoassay for blood cortisol measurement, determine the effect of storage on the radioimmunoassay, and determine the impact of any differences on clinical decisions. Methods The study included 54 client owned dogs undergoing adrenal function testing. Fresh plasma or serum samples (n = 170) were assayed for cortisol using radioimmunoassay (RIA1). Samples (n = 196) were also frozen and stored in batches, and assayed by chemiluminescence and radioimmunoassay (RIA2). Results Overall, there was a strong correlation (r² = 0.967, P < 0.001) between RIA2 and chemiluminescence concentrations without significant difference between means. Strong correlations were present for RIA2 and chemiluminescence at concentration subgroups of > 400 nmol/L (r² = 0.869, P < 0.001), < 100 nmol/L (r² = 0.790, P < 0.001), and < 40 nmol/L (r² = 0.738, P < 0.001). Significant differences between means were present for RIA2 and chemiluminescence concentrations in the < 100 nmol/L, and < 40 nmol/L (P < 0.001) groups. Despite a significant difference in RIA1 and RIA2 results overall, there was no significant difference between RIA1 and RIA2 for any of the concentration groups. In seven cases, discrepant RIA2 and chemiluminescence results may have altered clinical decisions. Conclusions Although RIA and chemiluminescence cortisol concentrations appear highly correlated, a significant difference may exist for concentrations less than 100 nmol/L in stored canine sera. Results of chemiluminescence cortisol assays should be interpreted with caution unless the specific assay method in the laboratory has been adequately validated in dogs.     

土壤脲酶活性两种测定方法的比较

利用比色法和扩散法对金强河地区和甘肃农业大学校园内的土壤脲酶活性进行了分析。结果表明：比色法和扩散法测定的土壤脲酶活性平均值分别为（0．531±0．207）mg／g和（0．446±0．204）mg／g,两种测定方法的结果和变异系数差异均不显著（P〉0．05）。但从试验操作方面考虑,扩散法操作比较简单、所用药品种类少、时间短,是一种比较可行的土壤脲酶活性测定方法。     

美洲南瓜(Cucurbita pepo)种皮发育形态观察及其相关酶活性测定

采用组织整体染色和石蜡切片相结合的方法对美洲南瓜有壳品种04LAg-26-2和无壳品种04LAg-26-28种皮发育形态结构比较观察。结果表明,有壳美洲南瓜种皮是由表皮层、下皮层、厚壁组织层、软组织层和绿色组织层构成,无壳裸仁南瓜的形成主要是授粉后18 d种皮发育过程中厚壁组织中的细胞变形、瓦解,缺少木质素的积累而导致了下皮层、厚壁组织层的缺失;对2个品种的种皮、叶片、叶柄和瓜囊不同组织中木质素合成相关酶活性测定结果表明,无壳品种04LAg-26-28在授粉18 d后,种皮中苯丙氨酸解氨酶(PAL)、肉桂酸-4-羟基化酶(C₄H)、4-香豆酸-辅酶A连接酶(4CL)、肉桂醇脱氢酶(CAD)和过氧化物酶(POD)活性明显降低,而叶片、叶柄和瓜囊中这几种酶活性在授粉18 d 后,呈逐渐增加的趋势。     

Comparison of the VIDAS and IMMULITE‐2000 methods for cortisol measurement in canine serum

Background: Serum cortisol concentration is often measured in dogs for the diagnosis and monitoring of adrenal disease. An enzyme‐linked fluorescent assay (VIDAS method) on the MiniVidas analyzer has been validated for the measurement of cortisol concentration in human serum and could have applications for canine samples. Objective: The aim of this study was to compare canine cortisol results obtained using the VIDAS method with those obtained using the IMMULITE‐2000 immunoassay, which has previously been validated for canine serum. Methods: The concentration of cortisol in 40 canine serum samples was determined concurrently with the VIDAS and IMMULITE methods, the latter as the reference method. Pearson's correlation coefficient, linear, and Deming regression analyses and Bland–Altman analysis were used to compare the 2 methods. Acceptability of the new method was judged using a medical decision chart (MEDx chart). Results: Cortisol concentrations obtained with the IMMULITE method ranged from 23.1 to 1380 nmol/L. Correlation (r=.977) and simple linear regression (slope=1.0722, confidence interval [CI] 0.996–1.148; intercept=?4.799, CI ?42.838 to 33.240) revealed no proportional or constant error. Based on Deming regression and a Bland–Altman plot the 2 methods gave comparable results. The MEDx chart indicated that performance of the new method was good at decision limits of 40, 132, and 480 nmol/L. Conclusion: Results of the VIDAS method were comparable to those of the IMMULITE‐2000 reference method such that the VIDAS may be used as an alternative assay to evaluate serum cortisol concentration in dogs for the diagnosis of adrenal disease.     

A comparison of uniaxial and triaxial accelerometers for the assessment of physical activity in dogs

The present study compares the outputs of uniaxial accelerometer (UA) and triaxial accelerometer (TA) to determine whether UAs can be used instead of TAs in estimating physical activity (PA) in domestic dogs (N = 79). The PA of the dogs was measured simultaneously by a UA and a TA attached to regular collars under 3 different conditions: unstructured activity in the kennels (kennel activity); and 2 structured on-lead activities of different intensities on a designated pathway (walking and trotting). The study finds that UA consistently detected significantly more steps. Kennel activity showed the largest differences between the accelerometers, and trotting showed the least. Dogs in the heaviest body weight category showed the best correlation between the devices and the least differences between the accelerometers across the 3 activities. The limits of agreement were wide in all activities. Significantly higher agreements were associated with lower step counts in kennel activity and higher step counts in walking and trotting. The results show that these 2 types of accelerometers cannot be used interchangeably. Confounding factors, such as body weight, must be considered in future analysis of accelerometer outputs and selection of devices for different contexts. Optimal interpretation of objectively measured PA is paramount in supporting future dog fitness, health, and welfare studies.     

RESEARCH PAPER: Comparison of two different methods for physiologic dead space measurements in ventilated dogs in a clinical setting

ObjectiveTo compare physiologic dead space (V_D) and physiologic dead space to tidal volume (V_T) ratio (V_D/V_T) obtained by an automated single breath test for carbon dioxide (CO₂) (method SBT) and a manual calculation (method MC) in ventilated healthy dogs.Study designProspective clinical study.AnimalsTwenty client‐owned dogs, ASA I and II undergoing anaesthesia for clinical purposes.MethodsFollowing pre‐medication, induction of anaesthesia, and intubation of the trachea, intermittent positive pressure ventilation was commenced. Mixed expired CO₂ partial pressure (PēCO₂) was measured by two methods: method MC by analysis, using an infrared capnograph, of the expired gas collected in a mixing box and method SBT which calculated it automatically by a device consisting of a mainstream capnograph and a pneumotachograph. At four time points arterial partial pressure of CO₂ (PaCO₂) was measured. Physiologic dead space variables (V_D and V_D/V_T) were calculated manually (method MC) or automatically (method SBT) using the Bohr–Enghoff equation.Method MC and SBT were compared using Bland–Altman plots and linear regression. Intra‐class correlation coefficient (ICC) was used to measure consistency of each method.ResultsFour measurement pairs were obtained in all 20 dogs for method SBT and MC. The bias was ?1.15 mmHg, 7.97 mL and 0.02 for PēCO₂, V_D and V_D/V_T, respectively. Linear regression analysis revealed a correlation coefficient (r²) of 0.79, 0.94, and 0.83 for PēCO₂, V_D and V_D/V_T, respectively. The ICC revealed an excellent consistency for both methods.ConclusionsThe single breath test (SBT) can be used for clinical evaluation of V_D and V_D/V_T in anaesthetized ventilated dogs.Clinical relevanceThrough measuring V_D and V_D/V_T important information about lung ventilation can be obtained and the SBT is an easy method to use for this purpose.     

Comparison of 4 fixation and staining methods for the cytologic evaluation of ear canals with clinical evidence of ceruminous otitis externa

BACKGROUND: Swab cytology of ear canals is one of the most useful and rapid methods to assess the presence of external ear infections. Smears are generally stained with rapid Romanowsky-type stains, with or without prior heat fixation. OBJECTIVES: The aim of this study was to compare 4 different methods of fixation and staining of ear swab cytology samples from dogs. METHODS: Eight dogs with otitis externa were selected from a dermatology referral population. A cotton swab was used to obtain ceruminous material from 12 ear canals. Four smears of each swab were prepared on glass slides (randomly identified as A, B, C, or D) and air-dried for cytologic examination. Samples marked A were stained with Dip Quick (Jorgensen Laboratories Inc, Loveland, CO, USA) after heat fixation; samples marked B were stained without heat fixation; samples marked C were heat-fixed and dipped only in the counterstain (the blue reagent) of Dip Quick; and samples marked D were dipped only in the counterstain, without heat fixation. Ten high-power fields (hpf; X100 oil immersion objective) in each slide were evaluated by 2 observers, and total numbers of keratinocytes, yeast, bacteria, and neutrophils were counted. Statistical comparison was performed using an ANOVA model applied after verifying the normal distribution of the data, and using nonparametric sign tests and Wilcoxon signed rank tests. RESULTS: No statistically significant differences were observed in the numbers of keratinocytes, yeast, bacteria, or neutrophils among the 4 staining methods (P > .05), although significant interobserver differences were found. CONCLUSION: We conclude that heat fixation does not improve the quality of ceruminous ear swab samples for cytologic evaluation, and propose a 1-step dip in the blue reagent alone as a rapid method of staining samples from canine ear canals.     

Surgery: Comparison of three closure methods and two absorbable suture materials for closure of jejunal enterotomy incisions in healthy dogs

Summary

The macroscopic and histological appearance of jejunal antimesenteric incisions approximated with two different absorbable suture materials (monofilament versus multifilament) and three closure techniques (appositional single layer, crushing single layer, and double layer) were compared in healthy dogs at 14 or 28 days, postoperatively. No significant differences between the two suture materials were observed for most of the macroscopic or histological variables. However, the monofilament suture material caused significantly more fibrous tissue reaction in the muscular layer of the jejunum than did the multifilament suture material. Of the three enterotomy closure techniques used in this study, the appositional single‐layer method proved to be the best. The double‐layer closure method caused a significant decrease in the incisional circumference, the relative circumference, and volume of the jejunum, and a significant increase in jejunal wall thickness. Our findings suggest that canine jejunal enterotomy incisions can be closed using an appositional suture pattern with relatively rapidly absorbable monofilament suture material. The use of double‐layer suture patterns for closure of jejunal enterotomy incisions should be avoided because the size of the intestinal lumen may be reduced.     

Comparison of a semiquantitative point-of-care assay for the detection of canine microalbuminuria with routine semiquantitative methods for proteinuria

Background: It has been speculated that renal disease can be identified through the detection and quantification of microalbuminuria, however, reliable measurement of albuminuria in any quantity can be challenging. Recently, a new point‐of‐care immunoassay was validated for the specific detection of microalbuminuria and early renal disease in dogs. Objectives: The goal of this study was to determine if measurement of microalbuminuria by the point‐of‐care immunoassay correlated with results from routine semiquantitative methods for detecting proteinuria in dogs. Methods: One hundred and thirty‐eight urine samples, from 133 different dogs, submitted for urinalysis to the Clinical Pathology Laboratory at the University of Missouri‐Columbia Veterinary Medical Teaching Hospital were eligible for the study. Samples that contained >20 RBC/high power field (hpf) or >20 WBC/hpf were excluded, as were samples with insufficient volume to complete all tests. All samples were evaluated with a urinary dipstick with or without a sulfosalicylic acid turbidimetric test, a urine protein:creatinine (UPC) ratio, and the immunoassay for microalbuminuria. Data were analyzed by the Spearman rank order correlation. Results: Microalbuminuria results correlated significantly with those of the dipstick (r= 0.715), sulfosalicylic acid test (r= 0.742), and UPC ratio (r= 0.830). Correlation between the immunoassay and UPC ratio was the same (r= 0.830) when only samples with trace or 1+ proteinuria by dipstick were analyzed (n = 51). Conclusions: The point‐of‐care immunoassay results for microalbuminuria correlated with the results of semiquantitative methods for detecting total proteinuria in dogs. Routine methods for canine proteinuria appear to be adequate for determining whether further testing for renal disease is warranted.     

3种检测牛白细胞源抗菌肽抗病毒活性方法的建立

为了建立牛白细胞源抗菌肽抗病毒活性的检测方法,采用半数组织细胞感染量(TCID₅₀)测定、荧光试验、MTT比色3种方法,用Vero ccl-81作为细胞模型,以猪水疱性口炎病毒(VSV)作为模式病毒,对牛白细胞源抗菌肽抗病毒活性进行评价。依据3种方法浓度水平测定的结果和由此结果推算出的抗病毒指标值建立回归模型,并对3种方法的稳定性进行试验。结果表明:抗菌肽浓度在0～62.5μg/mL时对Vero ccl-81细胞安全,在此浓度区间检测方法有效。在3种检测方法中,TCID₅₀法最准确,荧光试验特异直观迅速,MTT比色法准确又直观。TCID₅₀方法回归模型方程为:y=0.285x+1.890(R^2=0.928),荧光试验回归模型方程为:y=2.138x+6.136(R^2=0.903),CPE法回归模型方程为:y=10.229x+3.859(R^2=0.979),3种方法稳定性均较好。说明牛白细胞源抗菌肽基于TCID₅₀法、荧光试验、MTT比色法3种方法联合检测模型可以定量化地准确计算其抗病毒活性。     

不同种植年限条件下黄花蒿根际土壤微生物生物量、酶活性及真菌群落组成

试验采集未种植、种植1年、3年和5年的黄花蒿根际土壤,采用常规分析和Illumina MiSeq高通量测序技术,研究了土壤微生物生物量、酶活性及真菌群落组成。结果表明,在人工种植黄花蒿的土壤中,微生物生物量碳氮减少,碳氮比例改变;脱氢酶、脲酶和蔗糖酶活性降低,酸性磷酸酶活性增强;说明黄花蒿释放的化感物质选择性抑制了土壤微生物生长、繁殖和代谢。在不同种植年限的土壤中,主成分分析显示代表不同种植年限土壤真菌群落的点在坐标图中分布距离较远,表明它们的群落组成发生了显著变化(P<0.05)。此外,子囊菌门占土壤真菌的66.10%~95.28%,黄花蒿种植时间影响真菌门类和优势真菌的丰富度。在前20种优势真菌中,有14种共存于不同种植年限的土壤中,每种土壤中存在1~3种独有真菌,说明土壤是决定真菌种群组成的主导因素,又因种植黄花蒿而改变。在栽培1~5年的黄花蒿土壤中,优势菌株中出现蒿属的常见病菌——蒿白粉菌和艾菊柄锈菌,提高相应病害的发生风险。     

Comparison of three serotests for the detection of pseudorabies antibodies in pigs

The serum-neutralization test (SN), enzyme-linked immunosorbent assay (ELISA) and the radial immunodiffusion enzyme assay (RIDEA) were compared for the detection of pseudorabies (PRV) antibodies in swine sera. A total of 1285 serum samples were tested. All three tests were considered useful in determining the PRV antibody status of swine on a herd basis, but available evidence supports the continued use of SN as the definitive test because of possible false positive reactions associated with ELISA and RIDEA.