Validation of the shortened version of the Canine Behavioral Assessment and Research Questionnaire (C-BARQ) using participants from the Dog Aging Project.

The Canine Behavioral Assessment and Research Questionnaire (C-BARQ) is a 100-item owner-completed survey instrument used for assessing behavior and temperament of companion dogs. The shortened version of the C-BARQ (C-BARQ(S)) consists of 42 items of the long C-BARQ. We aimed to validate the shortened C-BARQ(S) by comparing it with the long questionnaire in the same human-dog pair. We examined data from a nationwide cohort of companion dogs enrolled in the large-scale longitudinal Dog Aging Project (DAP) study. Among 435 participating owners who completed both the long and shortened versions of the C-BARQ within 60 days of each other, agreement between individual questions of the long and shortened C-BARQ using an unweighted kappa statistic and percent agreement was examined. Associations between the two questionnaires for mean behavior and temperament domain scores and mean miscellaneous category scores were assessed using Pearson correlation coefficients. Of 435 dogs in the study, the mean (SD) age was 7.3 (4.3) years and 216 (50%) were female. Kappa values between the long and shortened C-BARQ for individual questions within the 14 behavior and temperament domains and a miscellaneous category ranged from fair to moderate (0.23 to 0.40 for 21 items and 0.41 to 0.58 for 26 items, respectively). Pearson correlation coefficients above 0.60 between both questionnaires for 12 of the 14 mean behavior and temperament domain scores and a category of miscellaneous items were observed. Kappa values for individual questions between the long and shortened C-BARQ ranged from fair to moderate and correlations between mean domain scores ranged from moderate to strong.

Clippers are superior to scissors in the collection of hair for chemical analysis in companion dogs: a Dog Aging Project preliminary study.

OBJECTIVE: To identify the safest, most efficient method for hair sample collection from companion dogs among clippers, scissors, and razors and to validate obtained samples with cortisol concentration analysis. ANIMALS: 25 healthy, privately owned dogs. METHODS: 2 hair samples were collected from each dog's ischiatic region with different implements (scissors, razors, or clippers). The collecting clinician completed a Hair Collection Questionnaire (HCQ) for each sample that compared subjective sample quality, time of collection, restraint needed, and patient experience. Each sample was evaluated by cortisol enzyme immunoassay. RESULTS: Clippers had higher overall HCQ scores than scissors, and scissors had higher HCQ scores than razors. Collection was faster for clippers than scissors, and scissors were faster than razors. There were no differences in sample quality between scissors and clippers, and sample quality was lower with razors. There was no difference in restraint needed or patient experience. Collection of long hair had higher HCQ scores than collection of medium and short hair. Collection of hair from dogs with an undercoat had higher HCQ scores than collection of hair from dogs without an undercoat. Dog size had no effect on HCQ score. Hair cortisol concentration did not vary between scissors or clippers (P = .111). Hair color and age did not affect hair cortisol concentration (P = .966 and P = .676, respectively). CLINICAL RELEVANCE: Clippers are recommended for hair sample collection from companion dogs. Scissors are an adequate alternative.

DNA methylation and chromatin accessibility predict age in the domestic dog.

Across mammals, the epigenome is highly predictive of chronological age. These "epigenetic clocks," most of which have been built using DNA methylation (DNAm) profiles, have gained traction as biomarkers of aging and organismal health. While the ability of DNAm to predict chronological age has been repeatedly demonstrated, the ability of other epigenetic features to predict age remains unclear. Here, we use two types of epigenetic information-DNAm, and chromatin accessibility as measured by ATAC-seq-to develop age predictors in peripheral blood mononuclear cells sampled from a population of domesticated dogs. We measured DNAm and ATAC-seq profiles for 71 dogs, building separate predictive clocks from each, as well as the combined dataset. We also use fluorescence-assisted cell sorting to quantify major lymphoid populations for each sample. We found that chromatin accessibility can accurately predict chronological age (R2 ATAC = 26%), though less accurately than the DNAm clock (R2 DNAm = 33%), and the clock built from the combined datasets was comparable to both (R2 combined = 29%). We also observed various populations of CD62L+ T cells significantly correlated with dog age. Finally, we found that all three clocks selected features that were in or near at least two protein-coding genes: BAIAP2 and SCARF2, both previously implicated in processes related to cognitive or neurological impairment. Taken together, these results highlight the potential of chromatin accessibility as a complementary epigenetic resource for modeling and investigating biologic age.

Dog size and patterns of disease history across the canine age spectrum: Results from the Dog Aging Project.

Age in dogs is associated with the risk of many diseases, and canine size is a major factor in that risk. However, the size patterns are complex. While small size dogs tend to live longer, some diseases are more prevalent among small dogs. In this study we seek to quantify how the pattern of disease history varies across the spectrum of dog size, dog age, and their interaction. Utilizing owner-reported data on disease history from a substantial number of companion dogs enrolled in the Dog Aging Project, we investigate how body size, as measured by weight, associates with the lifetime prevalence of a reported condition and its pattern across age for various disease categories. We found significant positive associations between dog size and the lifetime prevalence of skin, bone/orthopedic, gastrointestinal, ear/nose/throat, cancer/tumor, brain/neurologic, endocrine, and infectious diseases. Similarly, dog size was negatively associated with lifetime prevalence of ocular, cardiac, liver/pancreas, and respiratory disease categories. Kidney/urinary disease prevalence did not vary by size. We also found that the association between age and lifetime disease prevalence varied by dog size for many conditions including ocular, cardiac, orthopedic, ear/nose/throat, and cancer. Controlling for sex, purebred vs. mixed-breed status, and geographic region made little difference in all disease categories we studied. Our results align with the reduced lifespan in larger dogs for most of the disease categories and suggest potential avenues for further examination.

Analysis of 2,570 responses to Dog Aging Project End of Life Survey demonstrates that euthanasia is associated with cause of death but not age.

OBJECTIVE: The Dog Aging Project End of Life Survey was used to evaluate factors associated with manner of death (euthanasia vs unassisted death), including cause of death (CoD), reason for euthanasia (RFE) if performed, medical symptoms, old age characteristics, and perimortem quality of life (QoL). SAMPLE: Responses collected between the End of Life Survey launch (January 20, 2021) through December 31, 2021, from 2,570 participants whose dogs died. METHODS: Response frequencies were described. Associations between manner of death and medical symptoms or old age characteristics were evaluated using logistic regression. Factors associated with RFE were evaluated using multinomial regression. The effects of CoD, age at death, and QoL on the frequency of euthanasia as the manner of death were evaluated using multivariate logistic regression. RESULTS: 2,195 (85.4%) dogs were euthanized, and 375 (14.6%) experienced unassisted death. The most frequent owner-reported CoD was illness/disease (n = 1,495 [58.1%]). The most frequently reported RFE was pain/suffering (n = 1,080 [49.2% of those euthanized]). As age increased, RFE was more likely to be "poor QoL" than any other response. In a multivariate regression including CoD, chronologic age, and QoL, euthanasia as the manner of death was not significantly associated with age. CLINICAL RELEVANCE: Euthanasia was a common manner of death for dogs in the US. Compared with unassisted death, euthanasia was associated with CoD illness/disease, lower QoL scores, and the presence and number of medical symptoms and old age characteristics. Understanding factors associated with manner of death is important to veterinarians who care for dogs at the end of life.

Case report: Severe asymptomatic hypertriglyceridemia associated with long-term low-dose rapamycin administration in a healthy middle-aged Labrador retriever.

Rapamycin is an mTOR inhibitor that has been shown to extend the lifespan of laboratory model organisms. In humans, rapamycin is used at higher doses as an immunosuppressive medication to prevent organ rejection. Numerous adverse effects are seen with rapamycin treatment in humans, with one of the most common being dysregulation of lipid metabolism. In humans, this often manifests as mild to moderate serum lipid elevations, with a small subset developing extreme triglyceride elevations. This case report describes an eight-year-old, castrated male, clinically healthy Labrador retriever who developed severe hypertriglyceridemia associated with low-dose rapamycin administration over a six-month period. During this time, the dog was asymptomatic and displayed no other clinical abnormalities, aside from a progressive lipemia. Within 15 days of discontinuing rapamycin treatment, and with no targeted lipemic intervention, the dog's lipemia and hypertriglyceridemia completely resolved.

Lifetime prevalence of owner-reported medical conditions in the 25 most common dog breeds in the Dog Aging Project pack.

Introduction: Large scale data on the prevalence of diverse medical conditions among dog breeds in the United States are sparse. This cross-sectional study sought to estimate the lifetime prevalence of medical conditions among US dogs and to determine whether purebred dogs have higher lifetime prevalence of specific medical conditions compared to mixed-breed dogs. Methods: Using owner-reported survey data collected through the Dog Aging Project (DAP) Health and Life Experience Survey for 27,541 companion dogs, we identified the 10 most commonly reported medical conditions in each of the 25 most common dog breeds within the DAP cohort. Lifetime prevalence estimates of these medical conditions were compared between mixed-breed and purebred populations. The frequency of dogs for whom no medical conditions were reported was also assessed within each breed and the overall mixed-breed and purebred populations. Results: A total of 53 medical conditions comprised the top 10 conditions for the 25 most popular breeds. The number of dogs for whom no medical conditions were reported was significantly different (p = 0.002) between purebred (22.3%) and mixed-breed dogs (20.7%). The medical conditions most frequently reported within the top 10 conditions across breeds were dental calculus (in 24 out of 25 breeds), dog bite (23/25), extracted teeth (21/25), osteoarthritis (15/25), and Giardia (15/25). Discussion: Purebred dogs in the DAP did not show higher lifetime prevalence of medical conditions compared to mixed-breed dogs, and a higher proportion of purebred dogs than mixed-breed dogs had no owner-reported medical conditions. Individual breeds may still show higher lifetime prevalence for specific conditions.

A review of frailty instruments in human medicine and proposal of a frailty instrument for dogs.

Over the last few decades, frailty has become a pillar of research and clinical assessment in human gerontology. This complex syndrome, characterized by loss of physiologic reserves leading to decreased resilience to stressors, is of critical importance because it predicts higher risks of poor health outcomes, including mortality. Thus, identifying frailty among the elderly human population has become a key focus of gerontology. This narrative review presents current scientific literature on frailty in both humans and animals. The authors discuss the need for an accessible frailty instrument for companion dogs suitable for general use in veterinary medicine and the advances that would be facilitated by this instrument. A phenotypic frailty instrument for companion dogs, utilizing components that are easily collected by owners, or in the general practice setting, is proposed. The authors elaborate on the domains (physical condition, physical activity, mobility, strength, cognitive task performance, and social behavior), factors that will be included, and the data from the Dog Aging Project that inform each domain.

A masked, placebo-controlled, randomized clinical trial evaluating safety and the effect on cardiac function of low-dose rapamycin in 17 healthy client-owned dogs.

Introduction: Geroscience studies of low-dose rapamycin in laboratory species have identified numerous benefits, including reversing age-related cardiac dysfunction. Cardiovascular benefits have been observed in dogs with 10 weeks of treatment, raising questions about possible benefits and adverse effects of long-term use of low-dose rapamycin. The objectives of this study were to assess the impact of 6 months of low-dose rapamycin on echocardiographic indices of cardiac function in healthy dogs and to document the occurrence of adverse events. Methods: Seventeen client-owned dogs aged 6-10 years, weighing 18-36 kg, and without significant systemic disease were included in a prospective, randomized, placebo-controlled, masked clinical trial. Low-dose rapamycin (0.025 mg/kg) or placebo was administered three times per week for 6 months. Baseline, 6-month, and 12-month evaluation included physical examination, cardiology examination, and clinicopathology. Three-month evaluation included physical examination and clinicopathology. Owners completed online questionnaires every 2 weeks. Results: There were no statistically significant differences in echocardiographic parameters between rapamycin and placebo groups at 6 or 12 months. No clinically significant adverse events occurred. In 26.8% of the bi-weekly surveys owners whose dogs received rapamycin reported perceived positive changes in behavior or health, compared to 8.1% in the placebo group (p = 0.04). Discussion: While no clinically significant change in cardiac function was observed in dogs treated with low-dose rapamycin, the drug was well-tolerated with no significant adverse events.

Development and validation of a novel instrument to capture companion dog mortality data: the Dog Aging Project End of Life Survey.

OBJECTIVE: The researchers and clinicians within the Dog Aging Project (DAP), a longitudinal cohort study of aging in companion dogs, created and validated a novel survey instrument titled the End of Life Survey (EOLS) to gather owner-reported mortality data about companion dogs. SAMPLE: Bereaved dog owners who participated in the refinement, face validity assessment, or reliability assessment of the EOLS (n = 42) and/or completed the entire survey between January 20 and March 24, 2021 (646). PROCEDURES: The EOLS was created and modified by veterinary health professionals and human gerontology experts using published literature, clinical veterinary experience, previously created DAP surveys, and feedback from a pilot study conducted with bereaved dog owners. The EOLS was subjected to qualitative validation methods and post hoc free-text analysis to evaluate its ability to thoroughly capture scientifically relevant aspects of companion dogs' deaths. RESULTS: The EOLS was well received with excellent face validity as assessed by dog owners and experts. The EOLS had fair to substantial reliability for the 3 validation themes-cause of death (κ = 0.73; 95% CI, 0.5 to 0.95), perimortem quality of life (κ = 0.49; 95% CI, 0.26 to 0.73), and reason for euthanasia (κ = 0.3; 95% CI, 0.08 to 0.52)-and had no need for any substantial content alterations based on free-text analysis. CLINICAL RELEVANCE: The EOLS has proven to be a well-accepted, comprehensive, and valid instrument for capturing owner-reported companion dog mortality data and has the potential to enhance veterinarians' ability to care for the aging dog population by illuminating their understanding of companion dogs' end-of-life experiences.

Development and Validation of a Novel Instrument to Capture Companion Dog Mortality Data: The Dog Aging Project End of Life Survey.

Objective: The researchers and clinicians within the Dog Aging Project (DAP), a longitudinal cohort study of aging in companion dogs, created and validated a novel survey instrument titled the End of Life Survey (EOLS) to gather owner-reported mortality data about companion dogs. Sample: Bereaved dog owners who participated in the refinement, face validity assessment, or reliability assessment of the EOLS (n=42) and/or completed the entire survey between January 20 and March 24, 2021 (n=646). Procedures: The EOLS was created and modified by veterinary health professionals and human gerontology experts using published literature, clinical veterinary experience, previously created DAP surveys, and feedback from a pilot study conducted with bereaved dog owners. The EOLS was subjected to qualitative validation methods and post-hoc free-text analysis to evaluate its ability to thoroughly capture scientifically relevant aspects of companion dogs' death. Results: The EOLS was well-received with excellent face validity as assessed by dog owners and experts. The EOLS had fair to substantial reliability for the three validation themes: cause of death (kappa = 0.73; 95% CI [0.5-0.95]), perimortem quality of life (kappa = 0.49; 95% CI [0.26-0.73]), and reason for euthanasia (kappa = 0.3; 95% CI [0.08-0.52]) and had no need for any substantial content alterations based on free-text analysis. Clinical Relevance: The EOLS has proven to be a well-accepted, comprehensive, and valid instrument for capturing owner-reported companion dog mortality data and has the potential to enhance veterinarians' ability to care for the aging dog population by illuminating their understanding of companion dogs' end-of-life experiences.

Development and evaluation of a survey instrument to assess veterinary medical record suitability for multi-center research studies.

Here we describe the development and evaluation of a survey instrument to assess the research suitability of veterinary electronic medical records (EMRs) through the conduct of two studies as part of the Dog Aging Project (DAP). In study 1, four reviewers used the instrument to score a total of 218 records in an overlapping matrix of pairs to assess inter-rater agreement with respect to appropriate format (qualification), identification match (verification), and record quality. Based upon the moderate inter-rater agreement with respect to verification and the relatively large number of records that were incorrectly rejected the instrument was modified and more specific instructions were provided. In study 2, a modified instrument was again completed by four reviewers to score 100 different EMRs. The survey scores were compared to a gold standard of board-certified specialist review to determine receiver operating curve statistics. The refined survey had substantial inter-rater agreement across most qualification and verification questions. The cut-off value identified had a sensitivity of 95 and 96% (by reviewer 1 and reviewer 2, respectively) and a specificity of 82% and 91% (by reviewer 1 and reviewer 2, respectively) to predict gold standard acceptance or rejection of the record. Using just qualification and verification questions within the instrument (as opposed to full scoring) minimally impacted sensitivity and specificity and resulted in substantial time savings in the review process.

The Evaluation of the Diagnostic Value of a PCR Assay When Compared to a Serologic Micro-Agglutination Test for Canine Leptospirosis.

Objective: To evaluate PCR assay sensitivity and specificity compared to that of microscopic agglutination test (MAT) for diagnosis of canine leptospirosis. Electronic records search was performed to identify dogs with results for both PCR and MAT testing for leptospirosis. Methods: All dogs were clinically ill. Diagnosis of leptospirosis was defined as an unvaccinated dog with a positive MAT titer of ≥1:800 or a vaccinated dog or dog with an unknown vaccination status with a positive MAT titer of ≥1:1,600. Diagnosis of leptospirosis was excluded based on MAT titer <1:800 on both the initial and convalescent samples or an initial MAT titer <1:800 and an alternative definitive diagnosis. Results: Forty-nine samples (urine, n = 39; blood, n = 10) were evaluated. Leptospirosis was diagnosed in 17 dogs and excluded in 26 dogs. Urine PCR assay demonstrated sensitivity of 69.2%, specificity of 100%, positive predictive value of 100%, and negative predictive value of 86.6%. Blood PCR assay demonstrated sensitivity of 25%, specificity of 100%, positive predictive value of 100%, and negative predictive value of 25%. Overall PCR sensitivity was 52.4%, specificity was 100%, positive predictive value was 100%, and negative predictive value was 73.7%. Conclusions: PCR assay performed on urine or blood has high specificity and positive predictive value when compared to MAT for diagnosis of clinical canine leptospirosis. Sensitivity and negative predictive value are moderate to low, so PCR testing should be performed in conjunction with paired MAT testing for canine leptospirosis. Prior antibiotic therapy does not preclude the use of the PCR test.

Age and Physical Activity Levels in Companion Dogs: Results From the Dog Aging Project.

While there has been an abundance of studies on the important relationship between physical activity and age in both dogs and humans, studies on dogs have primarily focused on how a dog's biological characteristics, such as their weight, affect the age-activity relationship. To date, there is little knowledge about how this relationship may be associated with contextual- and owner-level characteristics. We leveraged a large and novel data set from the Dog Aging Project (DAP) to investigate the extent to which the age-activity relationship is associated with certain dog and owner characteristics, namely dog size, owner age, and the environment in which they live. Dogs are a unique model for aging research as they are exposed to similar social and environmental elements as humans but have a shorter life span, allowing researchers to observe their entire life course. We find that older dogs are less active than younger dogs; rural dogs are more active than suburban and urban dogs, especially at younger ages; and larger dogs are more active than smaller dogs. These findings are generally consistent with previous studies. However, a surprising finding is that older owners have more active dogs than younger owners. As one of the first studies to utilize the large survey data from the DAP, this study lays the foundation for future investigations to further understand and identify the biological, social, and environmental causes, as well as consequences, of aging.

An open science study of ageing in companion dogs.

The Dog Aging Project is a long-term longitudinal study of ageing in tens of thousands of companion dogs. The domestic dog is among the most variable mammal species in terms of morphology, behaviour, risk of age-related disease and life expectancy. Given that dogs share the human environment and have a sophisticated healthcare system but are much shorter-lived than people, they offer a unique opportunity to identify the genetic, environmental and lifestyle factors associated with healthy lifespan. To take advantage of this opportunity, the Dog Aging Project will collect extensive survey data, environmental information, electronic veterinary medical records, genome-wide sequence information, clinicopathology and molecular phenotypes derived from blood cells, plasma and faecal samples. Here, we describe the specific goals and design of the Dog Aging Project and discuss the potential for this open-data, community science study to greatly enhance understanding of ageing in a genetically variable, socially relevant species living in a complex environment.

Dog Models of Aging.

As the most phenotypically diverse mammalian species that shares human environments and access to sophisticated healthcare, domestic dogs have unique potential to inform our understanding of the determinants of aging. Here we outline key concepts in the study of aging and illustrate the value of research with dogs, which can improve dog health and support translational discoveries. We consider similarities and differences in aging and age-related diseases in dogs and humans and summarize key advances in our understanding of genetic and environmental risk factors for morbidity and mortality in dogs. We address health outcomes ranging from cancer to cognitive function and highlight emerging research opportunities from large-scale cohort studies in companion dogs. We conclude that studying aging in dogs could overcome many limitations of laboratory models, most notably, the ability to assess how aging-associated pathways influence aging in real-world environments similar to those experienced by humans.

Canine Cognitive Dysfunction (CCD) scores correlate with amyloid beta 42 levels in dog brain tissue.

Alzheimer's disease (AD) is a significant burden for human health that is increasing in prevalence as the global population ages. There is growing recognition that current preclinical models of AD are insufficient to recapitulate key aspects of the disease. Laboratory models for AD include mice, which do not naturally develop AD-like pathology during aging, and laboratory Beagle dogs, which do not share the human environment. In contrast, the companion dog shares the human environment and presents a genetically heterogeneous population of animals that might spontaneously develop age-associated AD-like pathology and cognitive dysfunction. Here, we quantitatively measured amyloid beta (Aß42 or Abeta-42) levels in three areas of the companion dog brain (prefrontal cortex, temporal cortex, hippocampus/entorhinal cortex) and cerebrospinal fluid (CSF) using a newly developed Luminex assay. We found significant positive correlations between Aß42 and age in all three brain regions. Brain Aß42 abundance in all three brain regions was also correlated with Canine Cognitive Dysfunction Scale score in a multivariate analysis. This latter effect remained significant when correcting for age, except in the temporal cortex. There was no correlation between Aß42 in CSF and cognitive scores; however, we found a significant positive correlation between Aß42 in CSF and body weight, as well as a significant negative correlation between Aß42 in CSF and age. Our results support the suitability of the companion dog as a model for AD and illustrate the utility of veterinary biobanking to make biospecimens available to researchers for analysis.

Veterinary Big Data: When Data Goes to the Dogs.

Dogs provide an ideal model for study as they have the most phenotypic diversity and known naturally occurring diseases of all non-human land mammals. Thus, data related to dog health present many opportunities to discover insights into health and disease outcomes. Here, we describe several sources of veterinary medical big data that can be used in research. These sources include medical records from primary medical care centers or referral hospitals, medical claims data from animal insurance companies, and datasets constructed specifically for research purposes. No data source provides information that is without limitations, but large-scale, prospective, longitudinally collected data from dog populations are ideal for further research as they offer many advantages over other data sources.

Reasons for Exclusion of Apparently Healthy Mature Adult and Senior Dogs From a Clinical Trial.

Background: Interventional clinical trials intended to maintain health in aging dogs are unusual and require particular attention to exclusion criteria. Objectives: To describe reasons for exclusion when a mature adult and senior canine population with normal health status was sought. Animals: Fifty six companion dogs nominated for a randomized controlled trial (RCT). Procedures: Exclusions occurred within Stage 1 (S1): owner-provided survey information; Stage 2 (S2): medical records review; and Stage 3 (S3): screening examination and within Owner, Dog, or Other factor categories. Results: Of 56 nominated dogs, 39 were excluded at S1 (n = 19), S2 (n = 5), and S3 (n = 15), respectively. Dogs were excluded for Owner (n = 4), Dog (n = 27), Other (n = 6), and concurrent (Owner + Dog; n = 2) factors. The most common exclusion period was S1 (n = 19), with weight outside the target range being the most common exclusion factor in that stage (n = 10). Heart murmurs were the second most common exclusion factor (S1: n = 1; S3: n = 5); suspected or confirmed systemic illness was third most common (S1: n = 2; S2: n = 3; S3: n = 2). Among dogs who passed S1 and S2 screening (n = 32), 15 dogs (48%) were excluded at S3, for heart murmur > grade II/VI (n = 5), cardiac arrhythmias (n = 2), and clinicopathologic abnormalities (n = 2). Conclusions and Clinical Relevance: Dogs nominated for a clinical trial for healthy mature adult and senior dogs were excluded for size, previous diagnoses, and newly discovered cardiac abnormalities. For future interventions in mature adult and senior dogs of normal health status, it is important to define expected age-related abnormalities to ensure that meaningful exclusion criteria are used.