Untargeted Analysis of Serum Metabolomes in Dogs with Exocrine Pancreatic Insufficiency.

Exocrine pancreatic insufficiency (EPI) is a malabsorptive syndrome resulting from insufficient secretion of pancreatic digestive enzymes. EPI is treated with pancreatic enzyme replacement therapy (PERT), but the persistence of clinical signs, especially diarrhea, is common after treatment. We used untargeted metabolomics of serum to identify metabolic disturbances associated with EPI and generate novel hypotheses related to its pathophysiology. Fasted serum samples were collected from dogs with EPI (n = 20) and healthy controls (n = 10), all receiving PERT. Serum metabolomes were generated using UPLC-MS/MS, and differences in relative metabolite abundances were compared between the groups. Of the 759 serum metabolites detected, 114 varied significantly (p < 0.05, q < 0.2) between dogs with EPI and healthy controls. Differences in amino acids (arginate, homoarginine, 2-oxoarginine, N-acetyl-cadaverine, and α-ketoglutaramate) and lipids (free fatty acids and docosahexaenoylcarnitine) were consistent with increased proteolysis and lipolysis, indicating a persistent catabolic state in dogs with EPI. Relative abundances of gut microbial metabolites (phenyllactate, 4-hydroxyphenylacetate, phenylacetyl-amino acids, catechol sulfates, and o-cresol-sulfate) were altered in dogs with EPI, consistent with disruptions in gut microbial communities. Increased kynurenine is consistent with the presence of intestinal inflammation in dogs with EPI. Whether these metabolic disturbances participate in the pathophysiology of EPI or contribute to the persistence of clinical signs after treatment is unknown, but they are targets for future investigations.

Longitudinal analysis of the rectal microbiome in dogs with diabetes mellitus after initiation of insulin therapy.

There have been numerous studies in humans and rodents substantiating the role of the gastrointestinal microbiome in the pathogenesis and progression of both type 1 and type 2 diabetes mellitus. Diabetes mellitus is a common endocrinopathy in dogs; however, little is known about the composition of the gut microbiome during the development and treatment of diabetes in this species. The objective of this pilot study was to characterize the gastrointestinal microbiome of dogs with diabetes mellitus at the time of diagnosis and over the first 12 weeks of insulin therapy and identify associations with glycemic control. Rectal swabs and serum for fructosamine measurement were collected from 6 newly diagnosed diabetic dogs at 2-week intervals for 12 weeks. Rectal samples were sequenced using 16S, ITS, and archaeal primers. Measures of alpha and beta diversity were assessed for changes over time; associations between absolute sequence variant (ASV) relative abundances and time and fructosamine concentration were identified using a microbiome-specific, multivariate linear effects model. No statistically significant changes over time were noted in alpha diversity and samples significantly grouped by dog rather than by time in the beta diversity analysis. However, multiple ASVs were negatively (Clostridium sensu stricto 1, Romboutsia, Collinsella) and positively (Streptococcus, Bacteroides, Ruminococcus gauveauii, Peptoclostridium) associated with time and two ASVs were positively associated with fructosamine (Enterococcus, Escherichia-Shigella). These changes in gastrointestinal microbial composition warrant further investigation of how they may relate to diabetes mellitus progression or control in dogs.

Toxidromes for Working Dogs.

Terrorist attacks with biological and chemical warfare agents are increasing in frequency worldwide. Additionally, hazardous chemical accidents, illicit drug laboratories and intentional poisonings are potential sites for exposure to working dogs. Working dogs play a crucial role in law enforcement, military and search and rescue teams. Their intelligence, agility and strength make them ideal partners to be deployed to these natural disaster sites, terrorist attacks and industrial accidents. This, unfortunately, leads to increasing exposure to chemical and biological weapons and other hazardous substances. First responders have little to no training in emergency care of working dogs and veterinarians have very little training on recognition of the clinical signs of many of these agents. In order to ensure a rapid medical response at the scene first responders and veterinarians need a primer on these agents. Identifying a specific agent amidst the chaos of a mass casualty event is challenging. Toxidromes are a constellation of clinical and/or laboratory findings that allow for rapid identification of the clinical signs associated with a class of toxin and have been helpful in human medical triage. Focusing on a class of agents rather than on each individual toxin, allows for more expedient administration of antidotes and appropriate supportive care. This article reviews toxidromes for the most common chemical weapons with a special emphasis on clinical signs that are specific (and different) for canines as well as appropriate antidotes for working canines. To our knowledge, there are no publications describing toxidromes for working dogs.

Case Report: Successful Intravenous Lipid Emulsion Therapy for Canine Amphetamine Toxicosis.

A 4-year-old, female-spayed, mixed breed dog, weighing 24.2 kg, was presented for acute ingestion of ~12.3 mg/kg of Adderall XRⓇ, an extended-release amphetamine medication. In dogs, the oral median lethal dose for amphetamines ranges anywhere from 9-11 mg/kg to 20-27 mg/kg. On presentation, the patient was agitated, tachycardic and hypertensive. Initial treatment was instituted with intravenous lipid emulsion (IVLE) therapy, and baseline and post-treatment amphetamine concentrations were quantified in serum and plasma. In both serum and plasma, post-IVLE concentrations of amphetamine were lower 1 h after treatment and IVLE was the only treatment instituted during this time. The dog improved significantly while in hospital and was discharged <24 h after presentation. This is the first known reported use of IVLE for treatment of amphetamine toxicosis with documented decreases in both serum and plasma amphetamine levels shortly after administration of IVLE.

Resolution of Severe Neurologic Signs Following Intravenous Lipid Emulsion Therapy in a Young Dog With a Portosystemic Shunt: Case Report.

A 5-month-old male intact Great Pyrenees was presented for an acute onset of severe neurologic signs (stupor, absent menace, intermittent head turn to the left). The patient's history included possible naproxen ingestion with a maximum ingested dose of 59 mg/kg, exceeding the reported dose of >50 mg/kg known to cause neurologic signs. Blood sampling for baseline bloodwork was performed, and intravenous lipid emulsion (ILE) was subsequently administered, for treatment of the suspected toxicosis. Due to severe and life-threatening neurologic signs, other methods of decontamination were contraindicated and unlikely to be effective; extracorporeal therapy was also unavailable. Complete resolution of neurologic signs occurred 30 min after completion of ILE therapy. At this time, the owners found the missing naproxen tablets after returning home and the bloodwork results returned revealing findings consistent with hepatic encephalopathy. The fasted blood ammonia concentration immediately prior to ILE administration was 702.1 µg/dL (reference interval, RI: 24-36 µg/dL) and decreased to 194.1 µg/dL 24 h later. In the first 24 h, the patient also received three doses of lactulose, N-acetylcysteine, and intravenous fluids. The patient was subsequently diagnosed with a single, large intrahepatic portosystemic shunt via computed tomography and underwent an endovascular coil embolization procedure. Given the rapid and dramatic improvement in severe neurologic signs after ILE therapy alone, it is strongly suspected that this treatment resulted in improvement of hepatic encephalopathy.

Current Rules and Regulations for Dogs Working in Assistance, Service, and Support Roles.

The legal landscape for dogs working in assistance, service, and support roles is complicated and contradictory. Regulations permit access to public places, allow subsets of dogs' emergency transport and treatment, provide elevated protections for K-9s and assistance animals from criminal acts, and make it a crime to fraudulently represent a service animal. Federal and state agencies provide different regulations for dogs to access public places. Identification and verification of the working animal are not standardized. Working dog legislation is a changing landscape that requires veterinarians to be up to date on laws and regulatory guidance.

Assistance, Service, Emotional Support, and Therapy Dogs.

This article focuses on the areas where harnessing the canine's trainability, mobility, and sociability enables their use for aiding and augmenting humans. This area, which is rapidly expanding, has provided life-changing solutions for persons affected by various impairments and disabilities (eg, visual, hearing, physical, mental).

Use of a Multimodal, Peer-to-Peer Learning Management System for Introduction of Critical Clinical Thinking to First-Year Veterinary Students.

Veterinary medical students need multiple thinking strategies, particularly critical thinking. We used a multimedia, peer review learning management system (CGScholar) to introduce a series of complex, realistic, case-based e-learning modules to help introduce critical thinking to 422 first-year veterinary students through instructor-designed clinical cases. Students developed and published on the CGScholar platform an analysis of a case and conducted anonymous peer reviews of each other's drafts. Instructors selected desirable characteristics of a student's activity to track and provide automatic feedback to students via an analytics dashboard and aster plot that allowed visualization of progress. The dashboard also enabled instructors to view the entire class's performance, highlighting students whose performance was lagging. Online interactions were supplemented by case-specific face-to-face workshop sessions. Our goal was to address the following questions: Does the addition of multimedia to a work (one's own or others') enhance people's ability to understand and convey the material? Does peer review (of one's own and others' work) lead to improvements in the writer's own work? Does the peer review process enhance the writer's understanding of what constitutes high-quality literature evidence? An anonymous student survey showed that experience was significantly more positive in the second and third year of implementation after inclusion of explicit guidance on the use of the rubric for peer review. Overall, 67% of students thought inclusion of multimedia enhanced their ability to communicate and 52% agreed multimedia enhanced their ability to understand their peers' analyses, but students were split on benefits to their understanding of high-quality literature.

Development and validation of a multivariate predictive model to estimate serum ionized calcium concentration from serum biochemical profile results in cats.

BACKGROUND: Measurement of serum ionized calcium is not always available in practice. Total calcium (tCa) might not be reliable for determination of calcium status in cats. OBJECTIVES: To predict serum ionized calcium concentration from signalment, biochemistry profile and T4, and compare predicted ionized calcium (piCa) to tCa. ANIMALS: A total of 1701 cats from two hospitals. METHODS: Cross-sectional study. Cats with serum ionized calcium, biochemistry profile and T4 available were screened over 6 years and included in the training set (569 cats) to create a multivariate adaptive regression splines model to calculate piCa. Diagnostic performances of tCa and piCa and its prediction interval (PI) were compared in 652 cats from the same institution (test set) and 480 cats from a different hospital (external set). RESULTS: The final model included tCa, chloride, albumin, cholesterol, creatinine, BUN, body condition score, GGT, age, and potassium. For hypercalcemia, piCa was highly specific (test set: 99.8%; confidence interval [CI]: 99.5-100; external set: 97%; CI: 95.3-98.7) but poorly sensitive (test set: 30.4%; CI: 18.3-42.4; external set: 42.5%; CI: 31.7-53.3). For hypocalcemia, piCa was also highly specific (test set: 81.6%; CI: 78-85; external set: 99.6%; CI: 99-100) but poorly sensitive (test set: 57.6%; CI: 50.6-64.6; external set: 0%). These diagnostic performances were comparable to those of tCa. The upper and lower limits of piCa PI had high sensitivity for detecting ionized hypercalcemia and hypocalcemia, respectively. CONCLUSIONS AND CLINICAL IMPORTANCE: Predicted ionized calcium is useful to confirm suspected hypercalcemia in cats and screen for hypercalcemia and hypocalcemia.

The Carle-Illinois (Urbana, Illinois USA) Transport Protocol for LEK9s: Guidelines for Emergency Medical Service Providers.

Three states and one county now allow Emergency Medical Services (EMS) providers to transport injured law enforcement K9s (LEK9s) as long as no human needs the ambulance at the time. Several other states either have pending legislation or are in discussions about this topic. As additional states ponder these laws, it is likely that the EMS transport of LEK9s will become legal in many states. In the wake of this legislation, a significant void was created. Currently, there are no published protocols for the safe transport of LEK9s by EMS providers. Additionally, the transport destination for these LEK9s is unlikely to be programmed into vehicle Global Positioning Systems. The authors of this report convened a Joint Task Force on Working Dog Care, consisting of veterinarians, EMS directors, EMS physicians, and LEK9 handlers, who met to develop a protocol for LEK9s being transported to a veterinary facility. The protocol covers the logistics of getting the LEK9 into the ambulance (eg, when the handler is or is not available), appropriate restraint, and the importance of prior arrangements with a veterinary emergency facility. A LEK9 hand-off form and a Transport Policy Form are provided, downloadable, and customizable for each EMS provider. This protocol provides essential information on safety and transport logistics for injured LEK9s. The hope is that this protocol will assist EMS providers to streamline the transport of an injured LEK9 to an appropriate veterinary facility.

The Carle-Illinois (Urbana, Illinois USA) Treatment Protocol for Law Enforcement K9s: Guidelines for Emergency Medical Services.

This document is a resource for Emergency Medical Services (EMS) treating an injured law enforcement K9 (LEK9) in the field and/or during transport by ambulance to a veterinary hospital. A Joint Task Force on Working Dog Care was created, which included veterinarians, EMS directors, EMS physicians, and canine handlers, who met to develop a treatment protocol for injured LEK9s. The protocol covers many major life-threatening injuries that LEK9s may sustain in the line of duty, and also discusses personnel safety and necessary equipment. This protocol may help train EMS providers to save the life of an injured LEK9.

Comparison of clinical findings between dogs with suspected anaphylaxis and dogs with confirmed sepsis.

OBJECTIVE To compare clinical signs, laboratory test results, and imaging findings between dogs with suspected anaphylaxis and dogs with sepsis. DESIGN Retrospective case-case study. ANIMALS 10 dogs with suspected anaphylaxis and 22 dogs with confirmed sepsis that met the criteria for systemic inflammatory response syndrome. PROCEDURES Medical records for dogs in each group were reviewed and data extracted regarding signalment; reason for hospital admission; physical examination findings; results of CBC, serum biochemical analysis, coagulation testing, cytologic examination, and microbial culture; and imaging reports. RESULTS All dogs in the anaphylaxis group fulfilled the criteria for systemic inflammatory response syndrome. Dogs in both groups had gastrointestinal signs, lethargy, mentation change, and bleeding abnormalities. Dogs with suspected anaphylaxis had a significantly higher eosinophil count and serum alanine aminotransferase activity and lower blood pH than dogs with sepsis. Dogs with sepsis had a significantly higher band neutrophil count, serum globulins concentration, and serum alkaline phosphatase activity and lower serum glucose concentration. Dogs in both groups had intracavitary free fluid and ultrasonographic findings of thickened intestines, gas or fluid-filled intestines, and a thickened gallbladder wall. CONCLUSIONS AND CLINICAL RELEVANCE Clinical signs, laboratory values, and imaging findings may be similar in dogs with sepsis or anaphylaxis. Given the marked difference in prognosis and treatment, early differentiation is important. Anaphylaxis should be considered if a septic nidus cannot be identified, and supportive care should be considered for such patients.

Effects of a transdermal lidocaine patch on indicators of postoperative pain in dogs undergoing midline ovariohysterectomy.

OBJECTIVE To determine the effects of a transdermal lidocaine patch (TLP) on indicators of postoperative pain in healthy dogs following ovariohysterectomy. DESIGN Randomized, blinded controlled trial. ANIMALS 40 healthy shelter-owned female dogs admitted to a student surgery program for ovariohysterectomy. PROCEDURES Dogs were randomly assigned to receive after ovariohysterectomy a 5-cm-wide strip of TLP applied topically on both sides of the incision, for the full length of the incision and a wound dressing (n = 19) or a placebo patch (nonmedicated wound dressing; 21). All dogs underwent midline ovariohysterectomy. Immediately afterward, dogs received 2 IM morphine injections, carprofen (SC, q 12 h for 2 days), and the assigned patch (left in place for 18 hours). Postoperative comfort was evaluated by use of the short form of the Glasgow Composite Measures Pain Scale and serum cortisol concentrations measured prior to premedication and 1, 2, 4, 6, 8, 10, and 18 hours after surgery. RESULTS No significant difference in pain scores or serum cortisol concentrations was identified between dogs that received the TLP and dogs that received a placebo patch after ovariohysterectomy. CONCLUSIONS AND CLINICAL RELEVANCE The TLP provided no additional analgesic benefit to dogs treated concurrently with recommended doses of morphine and carprofen following ovariohysterectomy. Additional studies are needed to investigate whether similar results might be achieved in dogs treated concurrently with other analgesics. (J Am Vet Med Assoc 2017;250:1140-1147).

Evaluation of assays for quantification of DNA in canine plasma as an indirect marker of NETosis.

BACKGROUND: Neutrophil extracellular traps (NET), consisting of a filamentous DNA/chromatin-histone scaffold originating from neutrophils are part of the innate immune response, may be released under a variety of inflammatory conditions and are associated with an increased risk for thrombosis. OBJECTIVES: The purpose of this study was to evaluate a SYTOX green fluorescence assay and a histone-DNA complex (hisDNA) ELISA for quantification of NET-related DNA in canine plasma. METHODS: The influence of variations in blood sample handling on assay results was tested. Accuracy of the SYTOX green fluorescence and the hisDNA ELISA was evaluated with dilutional linearity using serial dilutions. Interference was assessed by addition of purified bilirubin or hemoglobin. Precision was determined by calculating the intra- and inter-assay CV. RESULTS: Preanalytic sample handling did not influence DNA measurements by either assay. Citrate and EDTA plasma samples were equivalent. For the DNA fluorescence assay, dilutional linearity was poor due to autofluorescence, which was corrected by addition of canine plasma to the diluent. The presence of bilirubin and hemoglobin also increased autofluorescence, and resulted in falsely low concentrations of DNA. On the hisDNA ELISA, pigmentemia had no effect. CONCLUSIONS: Both assays as modified in this study are suitable for measuring DNA in canine EDTA or citrate plasma. However, performance of the fluorescence assay was impacted by pigmentemia, and it was less sensitive than the ELISA in detecting the presence of nucleosome material in the plasma.

Concentrations of thromboxane metabolites in feline urine.

OBJECTIVE To determine the predominant thromboxane (TX) metabolite in urine of healthy cats, evaluate whether the method of sample collection would impact concentration of that metabolite, and propose a reference interval for that metabolite in urine of healthy cats. ANIMALS 17 cats (11 purpose-bred domestic shorthair cats, 5 client-owned domestic shorthair cats, and 1 client-owned Persian cat). PROCEDURES All cats were deemed healthy on the basis of results for physical examination, a CBC, serum biochemical analysis, urinalysis, and measurement of prothrombin time and activated partial thromboplastin time. Voided and cystocentesis urine samples (or both) were collected. Aliquots of urine were stored at -80°C until analysis. Concentrations of TXB2, 11-dehydroTXB2, and 2,3 dinorTXB2 were measured with commercially available ELISA kits. Urinary creatinine concentration was also measured. RESULTS 11-dehydroTXB2 was the most abundant compound, representing (mean ± SD) 59 ± 18% of the total amount of TX detected. In all samples, the concentration of 11-dehydroTXB2 was greater than that of 2,3 dinorTXB2 (mean, 4.2 ± 2.7-fold as high). Mean concentration of 11-dehydroTXB2 for the 17 cats was 0.57 ± 0.47 ng/mg of creatinine. A reference interval (based on the 5% to 95% confidence interval) of 0.10 to 2.1 ng of 11-dehydroTXB2/mg of creatinine was proposed for healthy cats. CONCLUSIONS AND CLINICAL RELEVANCE In this study, 11-dehydroTXB2 was the major TX metabolite in feline urine. Measurement of this metabolite may represent a noninvasive, convenient method for monitoring in vivo platelet activation in cats at risk for thromboembolism.

Iron metabolism following intravenous transfusion with stored versus fresh autologous erythrocyte concentrate in healthy dogs.

OBJECTIVE To determine effects of IV transfusion with fresh (3-day-old) or stored (35-day-old) autologous erythrocyte concentrate on serum labile iron concentration, iron-binding capacity, and protein interaction with iron in dogs. ANIMALS 10 random-source healthy dogs. PROCEDURES Dogs were randomly assigned to receive autologous erythrocyte concentrate stored for 3 days (n = 5) or 35 days (5). One unit of whole blood was collected from each dog, and erythrocyte concentrates were prepared and stored as assigned. After erythrocyte storage, IV transfusion was performed, with dogs receiving their own erythrocyte concentrate. Blood samples were collected from each dog before and 5, 9, 24, 48, and 72 hours after transfusion. Serum was harvested for measurement of total iron, labile iron, transferrin, ferritin, hemoglobin, and haptoglobin concentrations. RESULTS For dogs that received fresh erythrocytes, serum concentrations of the various analytes largely remained unchanged after transfusion. For dogs that received stored erythrocytes, serum concentrations of total iron, labile iron, hemoglobin, and ferritin increased markedly and serum concentrations of transferrin and haptoglobin decreased after transfusion. CONCLUSIONS AND CLINICAL RELEVANCE Transfusion with autologous erythrocyte concentrate stored for 35 days resulted in evidence of intravascular hemolysis in healthy dogs. The associated marked increases in circulating concentrations of free iron and hemoglobin have the potential to adversely affect transfusion recipients.