Accuracy of oxygen delivery through bubble humidifiers and nasal catheters.

OBJECTIVE: To evaluate the equipment used for nasal insufflation of oxygen and determine its accuracy. STUDY DESIGN: Original study. METHODS: Oxygen delivery assemblies consisting of a flowmeter, bubble humidifier, oxygen delivery tubing and nasal insufflation catheters were assembled. Single and double catheter assemblies were made for four sizes of nasogastric feeding tubes (3.5 Fr, 5.0 Fr, 8.0 Fr and 10.0 Fr) resulting in 64 individual assemblies. A gas flow analyzer measured oxygen flow at the tip of the nasal catheter assemblies and from the pressure relief valve (PRV) of the bubble humidifiers. Statistical analyses were conducted to assess the functionality of assemblies. For functional assemblies, the accuracy of oxygen flow relative to the prescribed flow settings was determined. RESULTS: Catheter size was significantly associated with the functionality of assemblies. Probability (95% confidence interval) of 3.5 Fr, 5.0 Fr and 8.0 Fr assemblies being functional was estimated at 0.53 (0.14, 0.89), 0.83 (0.36, 0.98) and 0.98 (0.76, 0.99), respectively. All 10.0 Fr assemblies were functional. Functional assemblies, in general, consistently under-delivered the prescribed flow because a large portion of set flow was diverted through the bubble humidifier PRV. CONCLUSIONS: Leaks through the PRV cause significant diversion of oxygen prior to it reaching the catheter tips. Smaller patients are particularly susceptible, as small catheters limit oxygen delivery creating proportionally greater leaks through the PRV. CLINICAL RELEVANCE: It was not possible to accurately deliver oxygen because of leaks through the PRV. Targeting a specific outcome (e.g., oxyhemoglobin saturation > 94%, PaO2 80-120 mmHg; 11-16 kPa) and avoiding unnecessarily high fractions of inspired oxygen cannot be done if flow delivery cannot be accurately assured. One possible solution would be to use a bubble humidifier with a 6 psi PRV that does not leak prior to reaching the opening pressure.

Effects of Tylosin Administration Routes on the Prevalence of Antimicrobial Resistance Among Fecal Enterococci of Finishing Swine.

Antibiotics can be administered orally or parenterally in swine production, which may influence antimicrobial resistance (AMR) development in gut bacteria. A total of 40 barrows and 40 gilts were used to determine the effects of tylosin administration route on growth performance and fecal enterococcal AMR. The antibiotic treatments followed Food and Drug Administration label directions and were as follows: (1) no antibiotic (CON), (2) 110 mg tylosin per kg feed for 21 d (IN-FEED), (3) 8.82 mg tylosin per kg body weight through intramuscular injection twice daily for the first 3 d of each week for 3 weeks (IM), and (4) 66 mg tylosin per liter of drinking water (IN-WATER). Antibiotics were administered during d 0 to 21 and all pigs were then fed the CON diet from d 21 to 35. Fecal samples were collected on d 0, 21, and 35. Antimicrobial susceptibility was determined by microbroth dilution method. No evidence of route × sex interaction (p > 0.55) was observed for growth performance. From d 0 to 21, pigs receiving CON and IN-FEED had greater (p < 0.05) average daily gain (ADG) than those receiving IM, with the IN-WATER group showing intermediate ADG. Pigs receiving CON had greater (p < 0.05) gain-to-feed ratio (G:F) than IM and IN-WATER, but were not different from pigs receiving IN-FEED. Overall, enterococcal isolates collected from pigs receiving IN-FEED or IM were more resistant (p < 0.05) to erythromycin and tylosin than CON and IN-WATER groups. Regardless of administration route, the estimated probability of AMR to these two antibiotics was greater on d 21 and 35 than on d 0. In summary, IM tylosin decreased ADG and G:F in finishing pigs, which may be because of a response to the handling during injection administration. Tylosin administration through injection and feed resulted in greater probability of enterococcal AMR to erythromycin and tylosin compared with in-water treatment.

Invited review: Reproducible research from noisy data: Revisiting key statistical principles for the animal sciences.

Reproducible results define the very core of scientific integrity in modern research. Yet, legitimate concerns have been raised about the reproducibility of research findings, with important implications for the advancement of science and for public support. With statistical practice increasingly becoming an essential component of research efforts across the sciences, this review article highlights the compelling role of statistics in ensuring that research findings in the animal sciences are reproducible-in other words, able to withstand close interrogation and independent validation. Statistics set a formal framework and a practical toolbox that, when properly implemented, can recover signal from noisy data. Yet, misconceptions and misuse of statistics are recognized as top contributing factors to the reproducibility crisis. In this article, we revisit foundational statistical concepts relevant to reproducible research in the context of the animal sciences, raise awareness on common statistical misuse undermining it, and outline recommendations for statistical practice. Specifically, we emphasize a keen understanding of the data generation process throughout the research endeavor, from thoughtful experimental design and randomization, through rigorous data analysis and inference, to careful wording in communicating research results to peer scientists and society in general. We provide a detailed discussion of core concepts in experimental design, including data architecture, experimental replication, and subsampling, and elaborate on practical implications for proper elicitation of the scope of reach of research findings. For data analysis, we emphasize proper implementation of mixed models, in terms of both distributional assumptions and specification of fixed and random effects to explicitly recognize multilevel data architecture. This is critical to ensure that experimental error for treatments of interest is properly recognized and inference is correctly calibrated. Inferential misinterpretations associated with use of P-values, both significant and not, are clarified, and problems associated with error inflation due to multiple comparisons and selective reporting are illustrated. Overall, we advocate for a responsible practice of statistics in the animal sciences, with an emphasis on continuing quantitative education and interdisciplinary collaboration between animal scientists and statisticians to maximize reproducibility of research findings.

On accuracy and precision of flowmeters used for oxygen therapy in a veterinary teaching hospital.

OBJECTIVE: To determine the accuracy and precision of flowmeters used for oxygen therapy in a veterinary teaching hospital. STUDY DESIGN: An observational study. METHODS: A total of 50 flowmeters used for oxygen therapy were evaluated using Defender 530 gas flow analyzers to measure flow. For each flowmeter, a minimum of seven flow settings were tested in random order and in triplicate. Flow measured at ambient conditions was converted to standardized flow specifications (21.1 °C and 760 mmHg) and analyzed using general linear mixed models. Flowmeters were considered accurate at a given flow setting when the targeted mean flow was within the corresponding 95% confidence interval. Precision of flow was characterized based on the magnitude of variance component estimates. RESULTS: Flowmeters of 1.0, 3.5 and 8.0 L minute-1 were considered accurate across flow settings corresponding to their capacity range. Flowmeters of 7.0 and 15.0 L minute-1 were accurate at flow settings ≤2.0 L minute-1. For flow settings ≥3.0 L minute-1, average oxygen flow was consistently below reference values. Precision varied with the capacity of the flowmeter, ranked by decreasing precision as 1.0 > 3.5 > 8.0 > 7.0 > 15.0 L minute-1. CONCLUSIONS AND CLINICAL RELEVANCE: A flowmeter of the smallest maximum capacity within the desired flow range is more appropriate for smaller patients where accurate, precise flow delivery is needed. Although 15.0 L minute-1 flowmeters were accurate at flow settings ≤2.0 L minute-1, the graduated increments do not allow exact flow settings <0.5 L minute-1. Flowmeters of 15 L minute-1 capacity should be useful for high-flow oxygen delivery for which accuracy and precision are not critical.

Effect of cleaning status on accuracy and precision of oxygen flowmeters of various ages.

OBJECTIVE: To evaluate oxygen flowmeters for accuracy and precision, assess the effects of cleaning and assess conformity to the American Society for Testing Materials (ASTM) standards. STUDY DESIGN: Experimental study. METHODS: The flow of oxygen flowmeters from 31 anesthesia machines aged 1-45 years was measured before and after cleaning using a volumetric flow analyzer set at 0.5, 1.0, 2.0, 3.0, and 4.0 L minute-1. A general linear mixed models approach was used to assess flow accuracy and precision. RESULTS: Flowmeters 1 year of age delivered accurate mean oxygen flows at all settings regardless of cleaning status. Flowmeters ≥5 years of age underdelivered at flows of 3.0 and 4.0 L minute-1. Flowmeters ≥12 years underdelivered at flows of 2.0, 3.0 and 4.0 L minute-1 prior to cleaning. There was no evidence of any beneficial effect of cleaning on accuracy of flowmeters 5-12 years of age (p > 0.22), but the accuracy of flowmeters ≥15 years of age was improved by cleaning (p < 0.05). Regardless of age, cleaning increased precision, decreasing flow variability by approximately 17%. Nine of 31 uncleaned flowmeters did not meet ASTM standards. After cleaning, a different set of nine flowmeters did not meet standards, including three that had met standards prior to cleaning. CONCLUSIONS: Older flowmeters were more likely to underdeliver oxygen, especially at higher flows. Regardless of age, cleaning decreased flow variability, improving precision. However, flowmeters still may fail to meet ASTM standards, regardless of cleaning status. CLINICAL RELEVANCE: Cleaning anesthesia machine oxygen flowmeters improved precision for all tested machines and partially corrected inaccuracies in flowmeters ≥15 years old. A notable proportion of flowmeters did not meet ASTM standards. Cleaning did not ensure that they subsequently conformed to ASTM standards. We recommend annual flow output validation to identify whether flowmeters are acceptable for continued clinical use.

Inferential considerations for low-count RNA-seq transcripts: a case study on the dominant prairie grass Andropogon gerardii.

BACKGROUND: Differential expression (DE) analysis of RNA-seq data still poses inferential challenges, such as handling of transcripts characterized by low expression levels. In this study, we use a plasmode-based approach to assess the relative performance of alternative inferential strategies on RNA-seq transcripts, with special emphasis on transcripts characterized by a small number of read counts, so-called low-count transcripts, as motivated by an ecological application in prairie grasses. Big bluestem (Andropogon gerardii) is a wide-ranging dominant prairie grass of ecological and agricultural importance to the US Midwest while edaphic subspecies sand bluestem (A. gerardii ssp. Hallii) grows exclusively on sand dunes. Relative to big bluestem, sand bluestem exhibits qualitative phenotypic divergence consistent with enhanced drought tolerance, plausibly associated with transcripts of low expression levels. Our dataset consists of RNA-seq read counts for 25,582 transcripts (60% of which are classified as low-count) collected from leaf tissue of individual plants of big bluestem (n = 4) and sand bluestem (n = 4). Focused on low-count transcripts, we compare alternative ad-hoc data filtering techniques commonly used in RNA-seq pipelines and assess the inferential performance of recently developed statistical methods for DE analysis, namely DESeq2 and edgeR robust. These methods attempt to overcome the inherently noisy behavior of low-count transcripts by either shrinkage or differential weighting of observations, respectively. RESULTS: Both DE methods seemed to properly control family-wise type 1 error on low-count transcripts, whereas edgeR robust showed greater power and DESeq2 showed greater precision and accuracy. However, specification of the degree of freedom parameter under edgeR robust had a non-trivial impact on inference and should be handled carefully. When properly specified, both DE methods showed overall promising inferential performance on low-count transcripts, suggesting that ad-hoc data filtering steps at arbitrary expression thresholds may be unnecessary. A note of caution is in order regarding the approximate nature of DE tests under both methods. CONCLUSIONS: Practical recommendations for DE inference are provided when low-count RNA-seq transcripts are of interest, as is the case in the comparison of subspecies of bluestem grasses. Insights from this study may also be relevant to other applications focused on transcripts of low expression levels.

Short communication: On recognizing the proper experimental unit in animal studies in the dairy sciences.

Sound design of experiments combined with proper implementation of appropriate statistical methods for data analysis are critical for producing meaningful scientific results that are both replicable and reproducible. This communication addresses specific aspects of design and analysis of experiments relevant to the dairy sciences and, in so doing, responds to recent concerns raised in a letter to the editor of the Journal of Dairy Science regarding journal policy for research publications on pen-based animal studies. We further elaborate on points raised, rectify interpretation of important concepts, and show how aspects of statistical inference and elicitation of research conclusions are affected.

Insecticide Rotation Programs with Entomopathogenic Organisms for Suppression of Western Flower Thrips (Thysanoptera: Thripidae) Adult Populations under Greenhouse Conditions.

Western flower thrips, Frankliniella occidentalis (Pergande), is one of the most destructive insect pests of greenhouse production systems with the ability to develop resistance to a wide variety of insecticides. A common resistance management strategy is rotating insecticides with different modes of action. By incorporating entomopathogenic organisms (fungi and bacteria), which have discrete modes of action compared to standard insecticides, greenhouse producers may preserve the effectiveness of insecticides used for suppression of western flower thrips populations. The objective of this study was to determine how different rotation programs that include entomopathogenic organisms (Beauveria bassiana, Isaria fumosoroseus, Metarhizium anisopliae, and Chromobacterium subtsugae) and commonly used standard insecticides (spinosad, chlorfenapyr, abamectin, and pyridalyl) may impact the population dynamics of western flower thrips adult populations by means of suppression. Eight-week rotation programs were applied to chrysanthemum, Dendranthema x morifolium plants and weekly counts of western flower thrips adults captured on yellow sticky cards were recorded as a means to evaluate the impact of the rotation programs. A final quality assessment of damage caused by western flower thrips feeding on foliage and flowers was also recorded. Furthermore, a cost comparison of each rotation program was conducted. Overall, insecticide rotation programs that incorporated entomopathogenic organisms were not significantly different than the standard insecticide rotation programs without entomopathogenic organisms in suppressing western flower thrips adult populations. However, there were no significant differences among any of the rotation programs compared to the water control. Moreover, there was no differential effect of the rotation programs on foliage and flower quality. Cost savings of up to 34% (in US dollars) are possible when including entomopathogenic organisms in the rotation program. Therefore, by incorporating entomopathogenic organisms into insecticide rotation programs, greenhouse producers can decrease costs without affecting suppression, as well as diminish selection pressure on western flower thrips adult populations, which may avoid or delay resistance development.

Front of Pack Labels Enhance Attention to Nutrition Information in Novel & Commercial Brands.

OBJECTIVES: 1) To assess whether Front-of-Pack (FOP) nutrition labels garner attention more readily than more complete, mandated nutrition information (the Nutrition Facts Panel (NFP), required in the US), and 2) To determine whether label design characteristics, specifically, color coding and/or coding with facial icons, increase attention to the FOP label. METHODS: In two experiments, we tracked the allocation of attention while participants (n=125) viewed novel and commercial packages with varied FOP designs using a change detection methodology. RESULTS: We found empirical evidence that FOP labels are attended more often, and earlier, than the currently mandated NFP, and that this benefit is due both to its placement on the front of the package and to the design characteristics of the FOP. Specifically, the use of color in FOPs increased attention to the label, but there was no evidence that coding information via facial icons impacted attention. CONCLUSIONS: Our work supports a growing body of evidence supporting the use of FOP labels to attract attention to nutritional information. Findings may be relevant to inform policy decisions on labeling standards.

Ecotypes of an ecologically dominant prairie grass (Andropogon gerardii) exhibit genetic divergence across the U.S. Midwest grasslands' environmental gradient.

Big bluestem (Andropogon gerardii) is an ecologically dominant grass with wide distribution across the environmental gradient of U.S. Midwest grasslands. This system offers an ideal natural laboratory to study population divergence and adaptation in spatially varying climates. Objectives were to: (i) characterize neutral genetic diversity and structure within and among three regional ecotypes derived from 11 prairies across the U.S. Midwest environmental gradient, (ii) distinguish between the relative roles of isolation by distance (IBD) vs. isolation by environment (IBE) on ecotype divergence, (iii) identify outlier loci under selection and (iv) assess the association between outlier loci and climate. Using two primer sets, we genotyped 378 plants at 384 polymorphic AFLP loci across regional ecotypes from central and eastern Kansas and Illinois. Neighbour-joining tree and PCoA revealed strong genetic differentiation between Kansas and Illinois ecotypes, which was better explained by IBE than IBD. We found high genetic variability within prairies (80%) and even fragmented Illinois prairies, surprisingly, contained high within-prairie genetic diversity (92%). Using Bayenv2, 14 top-ranked outlier loci among ecotypes were associated with temperature and precipitation variables. Six of seven BayeScanFST outliers were in common with Bayenv2 outliers. High genetic diversity may enable big bluestem populations to better withstand changing climates; however, population divergence supports the use of local ecotypes in grassland restoration. Knowledge of genetic variation in this ecological dominant and other grassland species will be critical to understanding grassland response and restoration challenges in the face of a changing climate.

Intraocular pressure changes in equine athletes during endurance competitions.

OBJECTIVE: To assess intraocular pressure (IOP) in conditioned equine athletes and document changes with exercise. A secondary objective was to assess associations between IOP and heart rate, as well as with other subjective physical parameters. SAMPLE POPULATION: Horses were evaluated during 50 mile endurance ride competitions. Data were collected on 69 horses during five different competitions at three different locations with 59 horses ridden once, nine horses ridden in two competitions, and one horse ridden in three competitions for a total of 80 horse-ride combinations. PROCEDURES: Intraocular pressure was measured using a TonoVet(®) tonometer in both eyes of each horse prior to, at two time points during, and immediately after endurance competitions. Heart rates and subjective veterinary scores were recorded on ride cards at each time point. RESULTS: For horses with shorter finishing times (≤400 min), IOP decreased by at least 3.1 ± 0.9 mmHg (least square mean estimate ± SEM) from baseline to the end of endurance exercise (P < 0.007), although upward fluctuation was apparent during the ride. For horses with longer finishing times, IOP did not change significantly from baseline to the end of exercise. Responses also differed between horses awarded 'Best Condition' relative to other horses, whereby the latter, but not the former, showed an overall decrease in IOP by end of exercise relative to baseline (estimate decrease of 3.2 ± 0.6 mmHg; P < 0.001). There was no evidence for any association between IOP and heart rate, nor between IOP and subjective clinical scores. CONCLUSIONS: Intraocular pressure fluctuated in horses during endurance riding competitions.

Inferential implications of normalizing binomial proportions in a structural equation model: A simulation study motivated by feedlot data.

Most commercial software for implementation of structural equation models (SEM) cannot explicitly accommodate outcome variables of binomial nature. As a result, SEM modeling strategies of binomial outcomes are often based on normal approximations of empirical proportions. Inferential implications of these approximations are particularly relevant to health-related outcomes. The objective of this study was to assess the inferential implications of specifying a binomial variable as an empirical proportion (%) in predictor and outcome roles in a SEM. We addressed this objective first by a simulation study, and second by a proof-of-concept data application on beef feedlot morbidity to bovine respiratory disease (BRD). We simulated data on body weight at feedlot arrival (AW), morbidity count for BRD (Mb), and average daily gain (ADG). Alternative SEMs were fitted to the simulated data. Model 1 specified a directed acyclic causal diagram with morbidity fitted as a binomial outcome (Mb) and as a proportion (Mb_p) predictor. Model 2 specified a similar causal diagram with morbidity fitted as a proportion for both outcome and predictor roles within the network. Structural parameters for Model 1 were accurately estimated based on the nominal coverage probability of 95 % confidence intervals. In turn, there was poor coverage for most morbidity-related parameters under Model 2. Both SEM models showed adequate empirical power (>80 %) to detect parameters not equal to zero. Model 1 and Model 2 produced predictions that were reasonable from a management standpoint, as determined by calculating the root mean squared error (RMSE) through cross-validation. However, interpretability of parameter estimates in Model 2 was impaired due to the model misspecification relative to the data generation. The data application fitted SEM extensions, Model 1 * and Model 2 * , to a dataset from a group of feedlots in the Midwestern US. Models 1 * and 2 * included explanatory covariates, specifically percent shrink (PS), backgrounding type (BG), and season (SEA). Lastly, we tested if AW exerted both direct and BRD-mediated indirect effects on ADG using Model 2 * . In Model 1 * , mediation was not testable due to the incomplete path from morbidity as a binomial outcome through Mb_p as a predictor to ADG. Model 2 * supported a minor morbidity-mediated mechanism between AW and ADG, though parameter estimates were not directly interpretable. Our results indicate normal approximation to a binomial disease outcome in a SEM may be a viable option for inference on mediation hypotheses and for predictive purposes, despite limitations in interpretability due to inherent model misspecification.

Differential effects of temperature and mTOR and Wnt-planar cell polarity pathways on syndecan-4 and CD44 expression in growth-selected turkey satellite cell populations.

Satellite cells (SCs) comprise a heterogeneous population of muscle stem cells. Thermal stress during the first week after hatch alters proliferation, myogenesis, and adipogenesis of SCs of turkey pectoralis major (p. major) muscle via mechanistic target of rapamycin (mTOR) and wingless-type mouse mammary tumor virus integration site family/planar cell polarity (Wnt/PCP) pathways. Pivotal genes in mTOR and Wnt/PCP pathways are mTOR and frizzled-7 (Fzd7), respectively. The objective of this study was to determine the differential effects of thermal stress on SDC4 and CD44 expression in turkey p. major muscle SCs and how the expression of SDC4 and CD44 is modulated by the mTOR and Wnt/PCP pathways. Satellite cells were isolated from the p. major muscle of 1-week-old faster-growing modern-commercial (NC) turkeys and slower-growing historic Randombred Control Line 2 (RBC2) turkeys, and were challenged with hot (43°C) and cold (33°C) thermal stress for 72 h of proliferation followed by 48 h of differentiation. The NC line SCs were found to contain a lower proportion of SDC4 positive and CD44 negative (SDC4+CD44-) cells and a greater proportion of SDC4 negative and CD44 positive (SDC4-CD44+) cells compared to the RBC2 line at the control temperature (38°C) at both 72 h of proliferation and 48 h of differentiation. In general, at 72 h of proliferation, the proportion of SDC4+CD44- cells decreased with heat stress (43°C) and increased with cold stress (33°C) relative to the control temperature (38°C) in both lines, whereas the proportion of SDC4-CD44+ cells increased with heat stress and decreased with cold stress. In general, the expression of SDC4 and CD44 in the NC SCs showed greater response to both hot and cold thermal stress compared to the RBC2 cells. Knockdown of mTOR or Fzd7 expression increased the proportion of SDC4+CD44- cells while the proportion of SDC4-CD44+ cells decreased during differentiation with line differences being specific to treatment temperatures. Thus, differential composition of p. major muscle SCs in growth-selected commercial turkey may be resulted, in part, from the alteration in SDC4 and CD44 expression. Results indicate differential temperature sensitivity and mTOR and Wnt/PCP pathway responses of growth-selected SC populations and this may have long-lasting effect on muscle development and growth.

Examining the conspicuousness and prominence of two required warnings on OTC pain relievers.

The labeling of over-the-counter (OTC) drugs is critical to their safe and effective use, and certain warnings are meant to be read at the point of purchase (POP). Examples include (i) warnings that alert consumers to the fact that the package is not child-resistant and (ii) warnings that alert consumers to potential product tampering. U.S. law mandates these warnings be "conspicuous" and "prominent" so that it is likely that consumers will read them before leaving the store. Our objective was to quantify the relative prominence and conspicuousness of these warnings. Sixty-one participants reviewed the packages of 5 commercially available analgesics to evaluate the prominence and conspicuousness of these warnings. Evaluated data included (i) the time spent examining the warnings compared with other areas of the label (using a bright pupil eye tracker), (ii) the ability to recall information from the OTCs viewed, and (iii) the legibility of the warnings relative to other elements of the labels (as measured by ASTM D7298-06). Eye-tracking data indicated that warnings were viewed by fewer participants and for less time than other elements of the packages. Recall and legibility data also indicated that the warning statements compared unfavorably with other elements of the labels tested. Evidence presented in this study suggests that 2 required warnings on 5 different OTCs are not prominent or conspicuous when compared with other elements of tested labels.

Hierarchical Bayesian modeling of heterogeneous cluster- and subject-level associations between continuous and binary outcomes in dairy production.

The augmentation of categorical outcomes with underlying Gaussian variables in bivariate generalized mixed effects models has facilitated the joint modeling of continuous and binary response variables. These models typically assume that random effects and residual effects (co)variances are homogeneous across all clusters and subjects, respectively. Motivated by conflicting evidence about the association between performance outcomes in dairy production systems, we consider the situation where these (co)variance parameters may themselves be functions of systematic and/or random effects. We present a hierarchical Bayesian extension of bivariate generalized linear models whereby functions of the (co)variance matrices are specified as linear combinations of fixed and random effects following a square-root-free Cholesky reparameterization that ensures necessary positive semidefinite constraints. We test the proposed model by simulation and apply it to the analysis of a dairy cattle data set in which the random herd-level and residual cow-level effects (co)variances between a continuous production trait and binary reproduction trait are modeled as functions of fixed management effects and random cluster effects.

Toceranib phosphate in the management of canine insulinoma: A retrospective multicentre study of 30 cases (2009-2019).

BACKGROUND: Insulinomas are the most common tumour of the endocrine pancreas in dogs. These malignant tumours have a high metastatic rate and limited chemotherapeutic options. The multi-receptor tyrosine kinase inhibitor sunitinib malate has benefit in the treatment of metastatic insulinoma in people. Toceranib phosphate, an analogous veterinary agent, may provide benefit for dogs. METHODS: A retrospective study describing the extent and duration of clinical outcomes and adverse events (AEs) in dogs diagnosed with insulinoma and receiving toceranib. RESULTS: Records for 30 dogs diagnosed with insulinoma and having received toceranib were identified from a medical record search of five university and eight referral hospitals. The median progression-free interval and overall survival time were 561 days (95% confidence interval (CI): [246, 727 days]) and 656 days (95% CI: [310, 1045 days]), respectively. Of the dogs for which the canine Response evaluation criteria for solid tumours tool could be applied, the majority (66.7%) showed either a complete response, partial response or stable disease. Time to clinical progression was associated with prior intervention and type of veterinary practice. Larger dogs were at increased risk for disease progression and death. No novel AEs were reported. CONCLUSIONS: Most dogs diagnosed with insulinoma and receiving toceranib appeared to have a clinical benefit. Randomised, prospective studies are needed to better elucidate and objectively quantify the potential effect and survival benefit of toceranib therapy for management of insulinoma in dogs.

Investigating Salmonella enterica, Escherichia coli, and Coliforms on Fresh Vegetables Sold in Informal Markets in Cambodia†.

ABSTRACT: Vegetables in Cambodia are commonly sold in informal markets lacking food safety standards and controls. Current data on microbial contamination of vegetables in Cambodian informal markets are limited. The purpose of this study was to investigate Salmonella enterica and indicator organisms (Escherichia coli and coliforms) on the surface of fresh vegetables sold in informal markets in Cambodia. Samples of loose-leaf lettuce, tomatoes, and cucumbers were collected from 104 vendors at four informal markets in Battambang and Siem Reap provinces during the rainy and dry seasons. Detection methods for S. enterica were adapted from the U.S. Food and Drug Administration's Bacteriological Analytical Manual. Coliform and E. coli populations were quantified by plating onto E. coli/coliform count plates. S. enterica was most prevalent on lettuce during the dry season (56.5%, 95% confidence interval [CI] [41.0, 70.8]) than during the rainy season (15.4%, 95% CI [7.5, 29.1]), whereas no significant seasonal differences were apparent for tomatoes and cucumbers. Regardless of season, levels of S. enterica were highest on lettuce (5.7 log CFU/g, 95% CI [5.5, 5.9]), relative to cucumbers (4.2 log CFU/g, 95% CI [3.8, 4.6]) and tomatoes (4.3 log CFU/g, 95% CI [4.1, 4.6]). For E. coli, prevalence was higher during the rainy season (34.0%, 95% CI [25.4, 43.8]) than during the dry season (9.1%, 95% CI [4.9, 16.5]), with the highest prevalence estimated on lettuce. Coliform levels on lettuce and tomatoes were greater during the rainy season (6.3 and 5.3 log CFU/g, 95% CI [5.7, 6.8] and [4.7, 5.8], respectively) than during the dry season (5.2 and 3.9 log CFU/g, 95% CI [4.7, 5.7] and [3.4, 4.4], respectively). These results indicate seasonal patterns for microbial prevalence in lettuce and an overall high level of bacterial contamination on raw vegetables sold in Cambodian informal markets.

Prevalence of Salmonella enterica Isolated from Food Contact and Nonfood Contact Surfaces in Cambodian Informal Markets.

ABSTRACT: The lack of hygiene and sanitation practices and insufficient infrastructure in Cambodian informal markets may increase the risk of food contamination, specifically raw vegetables, which in turn may increase the chances of contracting a foodborne disease. The aims of this study in informal markets in Cambodia were (i) to quantify the prevalence of Salmonella enterica based upon differences in season of the year (rainy versus dry), surface types (food contact surfaces versus nonfood contact surfaces), and location of vendors within the market (inside versus outside) and (ii) to characterize S. enterica serotype prevalence. A total of 310 samples were screened for S. enterica prevalence following the U.S. Department of Agriculture guidelines, and results were confirmed by PCR assay. Whole genome sequencing was used to determine the serotype for each isolate in silico using SeqSero 1.0 on draft genomes. A total of 78 samples were confirmed positive for S. enterica. During the dry season, S. enterica was more prevalent on food contact surfaces than on nonfood contact surfaces (estimated probability of detection [confidence interval]: 0.41 [0.25, 0.59] and 0.17 [0.08, 0.32], respectively; P = 0.002), but no differences were apparent in the rainy season. No differences in S. enterica prevalence were found based on location within the market (P = 0.61). Sixteen S. enterica serotypes were detected across multiple surfaces. The most common S. enterica serotypes were Rissen (18 isolates), Hvittingfoss (11), Corvallis (10), Krefeld (8), Weltevreden (6), and Altona (6). Accurate data on the prevalence of S. enterica in informal markets are crucial for the development of effective surveillance and implementation of suitable intervention strategies at the domestic level, thus preventing foodborne illness.

Unmitigated Surgical Castration in Calves of Different Ages: Electroencephalographic and Neurohormonal Findings.

Castration is a common management procedure employed in North American cattle production and is known to cause a pain response. The present study was designed to investigate the effect of unmitigated surgical castration on the electroencephalography (EEG) responses and plasma substance P (SP) concentrations in calves of different ages under the same experimental conditions. Thirty male Holstein calves in three age categories [<6 weeks (6W); 3 months (3M); 6 months (6M); 10 calves per age group] were used in the study. Calves were subjected to a simulated castration session (SHAM) followed 24 h later by surgical castration (CAST) without analgesia. An EEG analysis was performed before the procedure (i.e., baseline), at treatment, and 0-5, 5-10, and 10-20 min post-treatment for both SHAM and CAST, respectively. Blood samples were collected immediately prior to both treatments (time 0) and again at 1, 2, 4, 8, and 12 h after both treatments. The EEG results showed a three-way interaction between treatment, age, and time for delta and beta absolute power, beta relative power, total power, and median frequency (p = 0.004, p = 0.04, p = 0.04, p = 0.03, and p = 0.008, respectively). Following CAST, EEG total power decreased, and median frequency increased relative to SHAM in 6W and 3M calves only following treatment. For 6W and 3M calves, delta and beta absolute power increased at CAST and at later time points relative to SHAM. Marginal evidence for two-way interactions was noted between time and treatment and between age and treatment on the concentration of SP (p = 0.068 and p = 0.066, respectively). Substance P concentrations decreased in CAST treatment compared to SHAM at the later times (8 h: p = 0.007; 12 h: p = 0.048); 6W calves showed lower SP concentration at CAST relative to SHAM (p = 0.017). These findings indicate variation in EEG responses and in SP concentrations following unmitigated surgical castration in calves and that these responses may be age specific. These EEG findings have implications for supporting the perception of the pain associated with surgical castration in young calves and emphasize the urgency of pain mitigation strategies during routine husbandry practices such as castration, as typically implemented in North American cattle management.

Unmitigated Surgical Castration in Calves of Different Ages: Cortisol Concentrations, Heart Rate Variability, and Infrared Thermography Findings.

The objective was to characterize physiological responses to unmitigated surgical castration in calves of varying ages. Thirty male Holstein calves of three ages [<6 w (6W); 3 m (3M); 6 m (6M); n = 10] underwent a simulated castration treatment (SHAM) followed 24 h later by castration (CAST). For both treatments, heart rate variability, eye temperature, and cortisol were measured over time from treatment to specified end points to capture the acute response period. Interactions between treatment and age (p = 0.035) and time and age (p < 0.001) were noted for cortisol. The 6W calves had lower cortisol compared to 6M calves at SHAM and CAST. Cortisol of 6W calves decreased from peak to pre-treatment levels faster than 6M calves. An interaction between time and age was reported in squared differences of inter-beat-intervals (RMSSD; p = 0.02) and high-frequency power (HFP; p = 0.05), whereby both responses decreased in 6W calves during the sampling period which was not seen in 3M and 6M calves. Average eye temperature (AET) differed by age (p = 0.0018) whereby 6W calves had lower AET than 6M calves (p = 0.0013) regardless of treatment and time. The findings suggest that responses to unmitigated surgical castration seem to be mediated by the autonomic nervous system in an age-related manner.