Association of serum and fecal microRNA profiles in cats with gastrointestinal cancer and chronic inflammatory enteropathy.

BACKGROUND: Differentiation of gastrointestinal cancer (GIC) from chronic inflammatory enteropathies (CIE) in cats can be challenging and often requires extensive diagnostic testing. MicroRNAs (miRNAs) have promise as non-invasive biomarkers in serum and feces for diagnosis of GIC. HYPOTHESIS/OBJECTIVES: Cats with GIC will have serum and fecal miRNA profiles that differ significantly from healthy cats and cats with CIE. Identify serum and fecal miRNAs with diagnostic potential for differentiation between cats with GIC and CIE as compared to healthy cats. ANIMALS: Ten healthy cats, 9 cats with CIE, and 10 cats with GIC; all client-owned. METHODS: Cats were recruited for an international multicenter observational prospective case-control study. Serum and feces were screened using small RNA sequencing for miRNAs that differed in abundance between cats with GIC and CIE, and healthy cats. Diagnostic biomarker potential of relevant miRNAs from small RNA sequencing and the literature was confirmed using reverse transcription quantitative real-time PCR (RT-qPCR). RESULTS: Serum miR-223-3p was found to distinguish between cats with GIC and CIE with an area under the curve (AUC) of 0.9 (95% confidence interval [CI], 0.760-1.0), sensitivity of 90% (95% CI, 59.6-99.5%), and specificity of 77.8% (95% CI, 45.3-96.1%). Serum miR-223-3p likewise showed promise in differentiating a subgroup of cats with small cell lymphoma (SCL) from those with CIE. No fecal miRNAs could distinguish between cats with GIC and CIE. CONCLUSION AND CLINICAL IMPORTANCE: Serum miR-223-3p potentially may serve as a noninvasive diagnostic biomarker of GIC in cats, in addition to providing a much needed tool for the differentiation of CIE and SCL.

Association of fecal and serum microRNA profiles with gastrointestinal cancer and chronic inflammatory enteropathy in dogs.

BACKGROUND: Reliable biomarkers to differentiate gastrointestinal cancer (GIC) from chronic inflammatory enteropathy (CIE) in dogs are needed. Fecal and serum microRNAs (miRNAs) have been proposed as diagnostic and prognostic markers of GI disease in humans and dogs. HYPOTHESIS/OBJECTIVES: Dogs with GIC have fecal and serum miRNA profiles that differ from those of dogs with CIE. AIMS: (a) identify miRNAs that differentiate GIC from CIE, (b) use high-throughput reverse transcription quantitative real-time PCR (RT-qPCR) to establish fecal and serum miRNA panels to distinguish GIC from CIE in dogs. ANIMALS: Twenty-four dogs with GIC, 10 dogs with CIE, and 10 healthy dogs, all client-owned. METHODS: An international multicenter observational prospective case-control study. Small RNA sequencing was used to identify fecal and serum miRNAs, and RT-qPCR was used to establish fecal and serum miRNA panels with the potential to distinguish GIC from CIE. RESULTS: The best diagnostic performance for distinguishing GIC from CIE was fecal miR-451 (AUC: 0.955, sensitivity: 86.4%, specificity: 100%), miR-223 (AUC: 0.918, sensitivity: 90.9%, specificity: 80%), and miR-27a (AUC: 0.868, sensitivity: 81.8%, specificity: 90%) and serum miR-20b (AUC: 0.905, sensitivity: 90.5%, specificity: 90%), miR-148a-3p (AUC: 0.924, sensitivity: 85.7%, specificity: 90%), and miR-652 (AUC: 0.943, sensitivity: 90.5%, specificity: 90%). Slightly improved diagnostic performance was achieved when combining fecal miR-451 and miR-223 (AUC: 0.973, sensitivity: 95.5%, specificity: 90%). CONCLUSIONS AND CLINICAL IMPORTANCE: When used as part of a diagnostic RT-qPCR panel, the abovementioned miRNAs have the potential to function as noninvasive biomarkers for the differentiation of GIC and CIE in dogs.

A retrospective study of 237 dogs hospitalized with suspected acute hemorrhagic diarrhea syndrome: Disease severity, treatment, and outcome.

BACKGROUND: Few studies have investigated management and outcome in dogs with acute hemorrhagic diarrhea syndrome (AHDS), and there is a paucity of data on dogs with concurrent signs of sepsis. OBJECTIVES: To report outcome in dogs with suspected AHDS according to disease severity and antimicrobial treatment, and to evaluate effect of fluid resuscitation on clinical criteria. ANIMALS: Two hundred thirty-seven dogs hospitalized with suspected AHDS. METHODS: Retrospective study based on medical records. Disease severity was evaluated using AHDS index, systemic inflammatory response syndrome (SIRS) criteria, and serum C-reactive protein (CRP) according to 3 treatment groups: No, 1, or 2 antimicrobials. RESULTS: Sixty-two percent received no antimicrobials, 31% received 1 antimicrobial, predominantly aminopenicillins, and 7% received 2 antimicrobials. At admission, median AHDS index was 13 (interquartile range, 11-15), which decreased significantly after the first day's hospitalization (P < .001) for all groups. Compared with no antimicrobials (7%), more dogs had ≥2 SIRS criteria in the antimicrobial groups (15% and 36%, respectively). C-reactive protein (CRP) correlated positively with AHDS index at hospitalization (P < .001). Across treatment groups, rehydration markedly reduced number of clinical SIRS criteria. Survival to discharge was 96%, lower for dogs receiving 2 antimicrobials (77%, P < .05). CONCLUSIONS AND CLINICAL IMPORTANCE: The majority of dogs hospitalized with suspected AHDS improve rapidly with symptomatic treatment only, despite signs of systemic disease on initial presentation. The often-used SIRS criteria might be a poor proxy for identifying dogs with AHDS in need of antimicrobial treatment, in particular when hypovolemic. The role of CRP in clinical decision-making or prognostication warrants further investigation.

Antimicrobial reduction measures applied in Danish pig herds following the introduction of the "Yellow Card" antimicrobial scheme.

Following introduction of the antimicrobial restrictive "Yellow Card Scheme" in summer 2010, a rapid decrease in the Danish national pig antimicrobial consumption was observed. The aims of this study were to (i) investigate which measures had been implemented to reduce the antimicrobial consumption according to farmers and veterinarians and (ii) where possible, investigate if said measures were reflected in the herds' antimicrobial purchase data. Based on national register data from VetStat and the Central Husbandry Register, the study population was selected among Danish pig herds which had decreased their annual antimicrobial consumption with ≥10% following the introduction of the Yellow Card Scheme comparing June 1, 2009-May 31, 2010 to June 1, 2010-May 31, 2011. Subsequently, questionnaire surveys of both farmers and veterinarians were carried out, resulting in responses from 179 farmers accounting for 202 herds (response ratio: 83%) and 58 veterinarians accounting for 140 herds. Prior to the introduction of the Yellow Card Scheme, 24% of the participating herds had an antimicrobial consumption for one or more age groups which exceeded the Yellow Card Scheme threshold values on antimicrobial consumption, while 50% of the herds had an antimicrobial consumption below the national average. The measures most frequently stated as having contributed to the antimicrobial reduction were increased use of vaccines (52% of farmers; 35% of the veterinarians), less use of group medication (44% of the farmers; 58% of the veterinarians) and staff education (22% of the farmers; 26% of the veterinarians). Reduced usage of antimicrobials for oral use accounted for 89% of the total reduction in antimicrobial use. Among the farmers, 13% also stated that change in choice of product had contributed to reducing their antimicrobial consumption. However, when analyzing purchase data, no general trend was seen towards a larger purchase of products with a higher registered dosage per kg animal compared to similar products. The findings of this study indicate that implementation of antimicrobial restrictive legislation at herd-level may lead to a variety of antimicrobial reducing initiatives in both herds with a high- and herds with a low previous level of antimicrobial consumption.

Reporting the national antimicrobial consumption in Danish pigs: influence of assigned daily dosage values and population measurement.

BACKGROUND: Transparent calculation methods are crucial when investigating trends in antimicrobial consumption over time and between populations. Until 2011, one single standardized method was applied when quantifying the Danish pig antimicrobial consumption with the unit "Animal Daily Dose" (ADD). However, two new methods for assigning values for ADDs have recently emerged, one implemented by DANMAP, responsible for publishing annual reports on antimicrobial consumption, and one by the Danish Veterinary and Food Administration (DVFA), responsible for the Yellow Card initiative. In addition to new ADD assignment methods, Denmark has also experienced a shift in the production pattern, towards a larger export of live pigs. The aims of this paper were to (1) describe previous and current ADD assignment methods used by the major Danish institutions and (2) to illustrate how ADD assignment method and choice of population and population measurement affect the calculated national antimicrobial consumption in pigs (2007-2013). RESULTS: The old VetStat ADD-values were based on SPCs in contrast to the new ADD-values, which were based on active compound, concentration and administration route. The new ADD-values stated by both DANMAP and DVFA were only identical for 48 % of antimicrobial products approved for use in pigs. From 2007 to 2013, the total number of ADDs per year increased by 9 % when using the new DVFA ADD-values, but decreased by 2 and 7 % when using the new DANMAP ADD-values or the old VetStat ADD-values, respectively. Through 2007 to 2013, the production of pigs increased from 26.1 million pigs per year with 18 % exported live to 28.7 million with 34 % exported live. In the same time span, the annual pig antimicrobial consumption increased by 22.2 %, when calculated using the new DVFA ADD-values and pigs slaughtered per year as population measurement (13.0 ADDs/pig/year to 15.9 ADDs/pig/year). However, when based on the old VetStat ADD values and pigs produced per year (including live export), a 10.9 % decrease was seen (10.6 ADDs/pig/year to 9.4 ADDs/pig/year). CONCLUSION: The findings of this paper clearly highlight that calculated national antimicrobial consumption is highly affected by chosen population measurement and the applied ADD-values.

Changes in group treatment procedures of Danish finishers and its influence on the amount of administered antimicrobials.

When treating groups of pigs orally, antimicrobials can be administered through either feed or water. During the last decade, the group treatment procedure for finishers has shifted from feed to water administration. We hypothesized that farms implementing this change in treatment procedure would increase their total amount of administered antimicrobials. Based on Danish national register data, we performed a retrospective cohort study with three groups. The cohort of primary interest (Cohort Change) consisted of 50 finisher farms which changed their group treatment procedure from feed administration to water administration between 2008 and 2009. In addition, we identified 221 farms where treatment was administered through feed (Cohort Feed), and another 553 farms where treatment was administered through water (Cohort Water). Both of these groups retained their original treatment procedure throughout the study period. Cohort Change experienced a significant increase in the total amount of prescribed antimicrobials between the years. This increase might be caused by the treatment of more pigs, since antimicrobials administered through the feed are mainly administered at the pen level, while antimicrobials administered in water are mainly administered at the section level. However, we cannot exclude that a change in clinical disease has influenced the amount of prescribed antimicrobials. No change was observed in the other two cohorts. Furthermore, the difference in the amount of prescribed antimicrobials between the years was significantly different in Cohort Change when compared to both Cohort Water and Cohort Feed. Results from this study demonstrate that farms changing their procedure of group treatment from feed administration to water administration may increase their overall use of antimicrobials.

Persistent Spatial Clusters of Prescribed Antimicrobials among Danish Pig Farms--A Register-Based Study.

The emergence of pathogens resistant to antimicrobials has prompted political initiatives targeting a reduction in the use of veterinary antimicrobials in Denmark, especially for pigs. This study elucidates the tendency of pig farms with a significantly higher antimicrobial use to remain in clusters in certain geographical regions of Denmark. Animal Daily Doses/100 pigs/day were calculated for all three age groups of pigs (weaners, finishers and sows) for each quarter during 2012-13 in 6,143 commercial indoor pig producing farms. The data were split into four time periods of six months. Repeated spatial cluster analyses were performed to identify persistent clusters, i.e. areas included in a significant cluster throughout all four time periods. Antimicrobials prescribed for weaners did not result in any persistent clusters. In contrast, antimicrobial use in finishers clustered persistently in two areas (157 farms), while those issued for sows clustered in one area (51 farms). A multivariate analysis including data on antimicrobial use for weaners, finishers and sows as three separate outcomes resulted in three persistent clusters (551 farms). Compared to farms outside the clusters during this period, weaners, finishers and sows on farms within these clusters had 19%, 104% and 4% higher use of antimicrobials, respectively. Production type, farm type and farm size seemed to have some bearing on the clustering effect. Adding these factors as categorical covariates one at a time in the multivariate analysis reduced the persistent clusters by 24.3%, 30.5% and 34.1%, respectively.

Weaner production with low antimicrobial usage: a descriptive study.

BACKGROUND: Health, productivity and antimicrobial use in the production of pigs are expected to be interrelated to some extent. Previous studies on register-based data have investigated these correlations with a subsequent large variation residing at the farm level. In order to study such farm factors in more detail we designed an elaborate interview-guide. By in-depth interviews of farmers with well-managed 7-30 kg (weaner) productions we sought to describe a set of common key-factors characterizing their management practices. Identification of such common practices could be used in follow-up projects, investigating whether identified factors really are characteristic for good-practicing famers. RESULTS: Eleven farms were selected for a farm visit and in-depth interview. Participating farms used less antimicrobials than the national median (8.2 animal daily doses/100 weaners/day), had a mortality below the national average (2.9%) and an average daily weight gain above the national average (443 g/day). Similarities were observed among participating farms, including the sectioning of farms, use of all-in-all-out procedures with subsequent cleaning, purchasing 7 kg weaners from only one source, as well as active participation in management by a committed farm owner. Most farmers had a specific point of focus in their management, and were convinced that this was the reason for their success. This included; feeding, treatment strategy, refurbishment of facilities and presence in the shed. CONCLUSION: According to register data, participating farms were alike; in the good league regarding use of antimicrobials, mortality and daily growth. However, on-farm interviews elucidated more heterogeneity among farmers than expected. Most of the farmers had a specific point of focus, which they considered to be crucial for their good results. These results indicate the importance of non-registerable factors, highlighting the value of qualitative study techniques in the understanding of human actions. Further studies on the effect of various farmer types are recommended.

Short-term survival and mortality rates in a retrospective study of colic in 1588 Danish horses.

BACKGROUND: Outcomes of colic treatment are of great interest to clinicians, horse owners and insurers. One commonly used criterion of success is the overall short-term survival rate. This is used as to compare treatments and to measure quality of veterinary care, but may be biased by demographic or social factors such as attitudes towards animal suffering and euthanasia. The aims of this study were to 1) describe and analyse characteristics in horses with signs of colic referred to the University Hospital for Large Animals (UHLA), University of Copenhagen, Denmark over a 10-year period and 2) to compare these rates with those published in other comparable studies. RESULTS: The overall survival rate for colic horses over the 10-year study period was 68% (confidence intervals (CI): 66-71%; 1087/1588). In the medical group, 1093 horses, short-term survival was 87% (CI: 85-89%). Thirty one % of referred horses were given diagnoses requiring surgical intervention (CI: 29-33%). In this group 32% of the horses were euthanized before surgery (CI: 28-36%; 159/495). Of the surgical cases 27% (CI: 23-31%) were euthanized or died during surgery. Of the horses that recovered from surgery 25% died or were euthanized (CI: 19-32%; 48/189), while 75% survived to discharge (CI: 68-81%). CONCLUSIONS: The short term survival rates of Danish horses with colic were similar or lower to those reported from other countries. Apart from variability of veterinary care, attitudes towards euthanasia vary among the countries, which may bias the outcomes. This study indicates that qualitative interview studies on owners' attitudes towards animal suffering and euthanasia need to be conducted. Our opinion is that survival rates are not valid as sole indicators of quality of care in colic treatment due to selection bias. If the survival rates are to be compared between hospitals, techniques or surgeons, prospective studies including mutually agreed-on disease severity scores and a predefined set of reasons for euthanasia are needed.