"Is free-roaming a key factor determining lifespan? An epidemiological study on the life expectancy of Turkish companion dogs".

Companion dog lifespan data has exclusively been studied in developed economies. Here we report results from n = 1312 privately owned Turkish companion dogs (Free-roaming and non Free-roaming) from an online survey analyzed through Kaplan-Meier analysis and Cox regression. Median survival time (MST) was 13 years. Most common causes of death were viral infections (n = 126), cancer (n = 60), and cardiovascular disease (n = 36). Desexing (χ2 = 31.6, P = 2E-8), being a mixed breed (χ2 = 6.4, P = 0.01), and regular preventative care (χ2 = 5.3, P = 0.02) significantly increased lifespan. Roaming freely significantly decreased lifespan (χ2 = 19.5, P = 1E-5). Dogs living in duplexes and single-family homes lived longer than dogs living in apartments and houses on acreage (χ2 = 10.5, P = 0.01). Owner income or education levels did not correlate with lifespan. In a Cox model, only desexing (HR = 0.478, P = 0.0006), living in a house on acreage (HR = 2.30, P = 0.0064) and being allowed to roam freely (HR = 1.59, P = 0.041) remained significant. Even though there are studies that contain information about dog demographics and mortality data outside of the western countries, to our knowledge, this is the first study of factors that influence companion dog lifespan in a middle income economy. While much of our findings correlate with those from developed economies, our sample also lets us study the effects of factors not commonly found in developed economies on dog lifespan, such as being allowed to roam freely.

Acarbose suppresses symptoms of mitochondrial disease in a mouse model of Leigh syndrome.

Mitochondrial diseases represent a spectrum of disorders caused by impaired mitochondrial function, ranging in severity from mortality during infancy to progressive adult-onset disease. Mitochondrial dysfunction is also recognized as a molecular hallmark of the biological ageing process. Rapamycin, a drug that increases lifespan and health during normative ageing, also increases survival and reduces neurological symptoms in a mouse model of the severe mitochondrial disease Leigh syndrome. The Ndufs4 knockout (Ndufs4-/-) mouse lacks the complex I subunit NDUFS4 and shows rapid onset and progression of neurodegeneration mimicking patients with Leigh syndrome. Here we show that another drug that extends lifespan and delays normative ageing in mice, acarbose, also suppresses symptoms of disease and improves survival of Ndufs4-/- mice. Unlike rapamycin, acarbose rescues disease phenotypes independently of inhibition of the mechanistic target of rapamycin. Furthermore, rapamycin and acarbose have additive effects in delaying neurological symptoms and increasing maximum lifespan in Ndufs4-/- mice. We find that acarbose remodels the intestinal microbiome and alters the production of short-chain fatty acids. Supplementation with tributyrin, a source of butyric acid, recapitulates some effects of acarbose on lifespan and disease progression, while depletion of the endogenous microbiome in Ndufs4-/- mice appears to fully recapitulate the effects of acarbose on healthspan and lifespan in these animals. To our knowledge, this study provides the first evidence that alteration of the gut microbiome plays a significant role in severe mitochondrial disease and provides further support for the model that biological ageing and severe mitochondrial disorders share underlying common mechanisms.

Lifetime prevalence of malignant and benign tumours in companion dogs: Cross-sectional analysis of Dog Aging Project baseline survey.

Although cancer is widely regarded as a major contributor to canine morbidity and mortality, its frequency in companion dogs has only infrequently been characterised. We analysed cross-sectional data from the baseline survey of owners of 27 541 living companion dogs enrolled in the Dog Aging Project as of 31 December 2020 to estimate the lifetime prevalence of malignant and benign tumours and several potentially-associated characteristics. Survey questions elicited information on history of 'cancer or tumors' including organ site and histologic type. Owners reported 819 malignant tumours (56% sited in the skin, muscle or other soft tissue) and 404 benign tumours (69% sited in the skin, muscle or other soft tissue). The lifetime prevalence of malignant tumours (29.7/1000 dogs) was approximately double the lifetime prevalence of benign tumours (14.7/1000 dogs). Lifetime prevalence of both malignant and benign tumours increased with dog age at survey completion. There were no statistically discernable differences in age-adjusted lifetime prevalence of malignant (prevalence ratio (PR) = 0.93 [95% confidence interval (CI) 0.82, 1.07] or benign tumours (PR = 1.10, 95% CI 0.91, 1.34) in mixed vs. purebred dogs. The lifetime prevalence of malignant tumours increased with increasing dog size class; compared to toy and small dogs, the age-adjusted PRs (95% CIs) for medium, standard, large, and giant dogs were 1.65 (1.28, 2.11), 2.92 (2.35, 3.64), 3.67 (2.92, 4.62) and 2.99 (1.23, 4.02), respectively. Similar though less pronounced patterns in relation to dog size class were observed for benign tumours. Ongoing prospective data collection will permit future studies on risk factors for canine tumour incidence.

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.

Lifespan of companion dogs seen in three independent primary care veterinary clinics in the United States.

BACKGROUND: The privately owned companion dog is an emerging model in comparative medicine, notably because it shares the human environment including its risk factors, is affected by many analogous age-related diseases, receives comparable medical care, and has excellent veterinary medical data available.Past studies of dog lifespan have used academic, corporate or insurance data. While independent primary care data exist for the UK, none have as of yet been published for the US. This study analyzed data from three independent primary care US veterinary hospitals and identified factors that influence lifespan and mortality in a cohort of n = 20,970 privately owned dogs using Kaplan-Meier survival estimators and Cox Proportional Hazards modelling, including body size as a covariate. RESULTS: As previously reported, body size was negatively correlated with lifespan. Gonadectomy was associated with a longer lifespan, with the effect being stronger in females than in males. This lifespan advantage was conserved in gonadectomized female dogs that lived to at least ages 5 and 8 years. We did not find significant differences in lifespan between purebred and mixed breed dogs; however, breeds with larger effective population sizes and/or lower inbreeding coefficients had median survival times 3-6 months longer than breeds with smaller effective population sizes or higher inbreeding coefficients, indicating that these measures of genetic diversity may be affecting breed lifespans. We also found that dog breeds belonging to the "Mountain" ancestral group had median survival times that were 3.5-4.6 years shorter than other purebred dog groups, which remained significant even when correcting for body size. CONCLUSIONS: Our findings show that it is possible to obtain and analyze data from independent veterinary clinics in the US, an approach that could be useful for studies of comparative epidemiology under the One Health and One Welfare paradigms. We also show that the lifespan effects of gonadectomy are not identical between the sexes and should be investigated separately by sex in future analyses. More research is needed to further clarify the influence of age at gonadectomy, as well as the factors leading to the observed differences in lifespan in the "Mountain" ancestral group and in dog breeds of varying inbreeding coefficients and effective population sizes.

Desexing Dogs: A Review of the Current Literature.

Background: Desexing dogs is promoted for population control, preventative healthcare, and behavior modification. Common methods are orchiectomy and ovariectomy/ovariohysterectomy. GnRH superagonist implants are available in some areas. Alternative methods like vasectomy and salpingectomy/hysterectomy are uncommon. The terminology used to describe desexing is inconsistent and contradictory, showing a need for the adaption of standardized terminology. Population Control: Surprisingly, empirical studies show no effects of desexing on population control in companion and shelter dogs despite desexing being consistently recommended in the literature. There is evidence for a population control effect in free-roaming dogs, where desexing also has benefits on zoonotic disease and bite risk. Population control in free-roaming dogs is mostly correlated with female, not male desexing. Health and Lifespan: Desexing affects numerous disease risks, but studies commonly neglect age at diagnosis and overall lifespan, age being by far the most important risk factor for most diseases. We argue that lifespan is a more important outcome than ultimate cause of death. A beneficial effect of desexing on lifespan is consistently demonstrated in females, while evidence for a beneficial effect in males is inconsistent. Studies are likely biased in desexing being a proxy for better care and desexed dogs having already lived to the age of desexing. Desexing reduces or eliminates common life-limiting diseases of the female reproductive system such as pyometra and mammary tumors, while no analogous effect exists in males. Disease risks increases across sexes and breeds include cruciate ligament rupture, various cancers, and obesity. Urinary incontinence risk is increased in females only. Various other disease risk changes show considerable variability between breeds and sexes. Behavioral Effects: Desexed males show reduced libido, roaming, conspecific mounting, and urinary marking frequency, as well as reduced male dog-directed aggression in a majority of males desexed for behavioral reasons. There is a detrimental effect on the risk and progression of age-related cognitive dysfunction. Desexed dogs may be less likely to cause bite injuries across sexes. The evidence for other effects such as human-directed aggression, human or object mounting, resource guarding, or shyness and anxiety is inconsistent and contradictory. There are few studies specific to females or individual breeds. Conclusions: The evidence for a beneficial effect of desexing is stronger in female than in male dogs; however, there is significant variation between breeds and sexes, and more research is needed to further elucidate these differences and to arrive at individualized evidence-based recommendations for clinical practice.

Cross species application of quantitative neuropathology assays developed for clinical Alzheimer's disease samples.

A major obstacle for preclinical testing of Alzheimer's disease (AD) therapies is the availability of translationally relevant AD models. Critical for the validation of such models is the application of the same approaches and techniques used for the neuropathological characterization of AD. Deposition of amyloid-ß 42 (Aß42) plaques and neurofibrillary tangles containing phospho-Tau (pTau) are the pathognomonic features of AD. In the neuropathologic evaluation of AD, immunohistochemistry (IHC) is the current standard method for detection of Aß42 and pTau. Although IHC is indispensable for determining the distribution of AD pathology, it is of rather limited use for assessment of the quantity of AD pathology. We have recently developed Luminex-based assays for the quantitative assessment of Aß42 and pTau in AD brains. These assays are based on the same antibodies that are used for the IHC-based diagnosis of AD neuropathologic change. Here we report the application and extension of such quantitative AD neuropathology assays to commonly used genetically engineered AD models and to animals that develop AD neuropathologic change as they age naturally. We believe that identifying AD models that have Aß42 or pTau levels comparable to those observed in AD will greatly improve the ability to develop AD therapies. Abbreviations: Alzheimer's disease (AD); amyloid ß 42 (Aß42); phospho-Tau (pTau); immunohistochemistry (IHC).

Risk Factors Associated with Lifespan in Pet Dogs Evaluated in Primary Care Veterinary Hospitals.

The objective of this population-based retrospective cohort study was to identify factors associated with lifespan in pet dogs evaluated at primary care veterinary hospitals. Dogs ≥3 mo of age that visited any of 787 US hospitals at least twice from January 1, 2010, through December 31, 2012, were included. Survival curves were constructed for dogs by reproductive status, breed, body size, and purebreed (versus mixed-breed) status. Multivariate Cox proportional hazard regression was performed to identify factors associated with lifespan. There were 2,370,078 dogs included in the study, of whom 179,466 (7.6%) died during the study period. Mixed-breed dogs lived significantly longer than purebred dogs, and this difference was more pronounced as body size increased. Controlling for other factors, dogs of either sex had a greater hazard of death over the study follow-up period if sexually intact rather than gonadectomized. For dogs who lived to 2 yr of age, the hazard of death decreased with increasing frequency of dental scaling. Our findings support previous reports of the impact of body size and gonadectomy on lifespan and provide new evidence in support of ultrasonic dental scaling and mixed breeding.

A randomized controlled trial to establish effects of short-term rapamycin treatment in 24 middle-aged companion dogs.

Age is the single greatest risk factor for most causes of morbidity and mortality in humans and their companion animals. As opposed to other model organisms used to study aging, dogs share the human environment, are subject to similar risk factors, receive comparable medical care, and develop many of the same age-related diseases humans do. In this study, 24 middle-aged healthy dogs received either placebo or a non-immunosuppressive dose of rapamycin for 10 weeks. All dogs received clinical and hematological exams before, during, and after the trial and echocardiography before and after the trial. Our results showed no clinical side effects in the rapamycin-treated group compared to dogs receiving the placebo. Echocardiography suggested improvement in both diastolic and systolic age-related measures of heart function (E/A ratio, fractional shortening, and ejection fraction) in the rapamycin-treated dogs. Hematological values remained within the normal range for all parameters studied; however, the mean corpuscular volume (MCV) was decreased in rapamycin-treated dogs. Based on these results, we will test rapamycin on a larger dog cohort for a longer period of time in order to validate its effects on cardiac function and to determine whether it can significantly improve healthspan and reduce mortality in companion dogs.

Asymptomatic heart valve dysfunction in healthy middle-aged companion dogs and its implications for cardiac aging.

Heart disease is the leading cause of death in the USA, accounting for about one in every four deaths. Age is the greatest risk factor for heart disease in both people and dogs; however, heart disease is generally not considered as a major cause of morbidity or mortality in dogs. As part of the preliminary selection process for a veterinary clinical trial, 40 companion dogs with no history of cardiac pathology that were at least 6 years old and weighed at least 18 kg underwent a cardiac screening using Doppler echocardiography. Eleven dogs from this cohort were diagnosed with valvular regurgitation by echocardiography, and seven of these cases were of sufficient severity to warrant exclusion from the clinical trial. In only one case was a heart murmur detected by auscultation. Serum alkaline phosphatase levels were significantly higher in the dogs with moderate to severe valvular regurgitation compared to the rest of the cohort. These observations suggest that asymptomatic degenerative valvular disease detectable by echocardiography, but not by a standard veterinary exam including auscultation, may be present in a significant fraction of middle-aged companion dogs, indicating a previously underappreciated similarity between human and canine aging. Further, these data suggest that companion dogs may be a particularly useful animal model for understanding mechanisms of age-related degenerative valve disease and for developing and testing interventions to ameliorate cardiac disease. Future studies should address whether dogs with asymptomatic valve disease are at higher risk for subsequent morbidity or early death.

Age-related retention of fiber cell nuclei and nuclear fragments in the lens cortices of multiple species.

PURPOSE: To determine the differences between species in the retention of lens fiber cell nuclei and nuclear fragments in the aging lens cortex and the relationship of nuclear retention to lens opacity. For this purpose old human, monkey, dog, and rat lenses were compared to those of three strains of mouse. We also investigated possible mechanisms leading to nuclear retention. METHODS: Fixed specimens of the species referred to above were obtained from immediate on site sacrifice of mice and rats, or from recently fixed lenses of other species, dogs, monkeys, and humans, obtained from collaborators. The retention of undegraded nuclei and nuclear fragments was graded 1-4 from histologic observation. All species lenses were examined microscopically in fixed sections stained with hematoxylin and eosin (H&E) or 4',6-diamidino-2-phenylindole (DAPI). Slit lamp observations were made only on the mice and rats before sacrifice and lens fixation. Values of 0 to 4 (clear lens to cataract) were given to degree of opacity. MRNA content in young versus old C57BL/6 mouse lenses was determined by quantitative PCR (qPCR) for DNase II-like acid DNase (DLAD) and other proteins. DLAD protein was determined by immunofluorescence of fixed eye sections. RESULTS: In old C57BL/6 and DBA mice and, to a lesser degree, in old CBA mice and old Brown Norway (BN) rats lenses were seen to contain a greatly expanded pool of unresolved whole nuclei or fragments of nuclei in differentiating lens fiber cells. This generally correlated with increased slit lamp opacities in these mice. Most old dog lenses also had an increase in retained cortical nuclei, as did a few old humans. However, a second rat strain, BNF1, in which opacity was quite high had no increase in retained nuclei with age nor did any of the old monkeys, indicating that retained nuclei could not be a cause of opacity in these animals. The nuclei and nuclear fragments were located at all levels in the outer cortex extending inward from the lens equator and were observable by the DAPI. These nuclei and nuclear fragments were seen from 12 months onward in all C57BL/6 and DBA/2 mice and to a lesser degree in the CBA, increasing in number and in space occupancy with increasing age. Preliminary results suggest that retention of nuclei in the C57BL/6 mouse is correlated with an age-related loss of DLAD from old lenses. CONCLUSIONS: A very marked apparently light refractive condition caused by retained cortical nuclei and nuclear fragments is present in the lens cortices, increasing with age in the three strains of mice examined and in one of two strains of rats (BN). This condition was also seen in some old dogs and a few old humans. It may be caused by an age-related loss of DLAD, which is essential for nuclear DNA degradation in the lens. However, this condition does not develop in old BNF1 rats, or old monkeys and is only seen sporadically in humans. Thus, it can not be a universal cause for age related lens opacity or cataract presence, although it develops concurrently with opacity in mice. This phenomenon should be considered when using the old mouse as a model for human age-related cataract.

Age-related cataract in dogs: a biomarker for life span and its relation to body size.

Clinical data from 72 dog breeds of varying size and life expectancy were grouped according to breed body mass and tested for prevalence at ages 4 to 5, ages 7 to 10, and lifetime incidence of non-hereditary, age-related cataract (ARC). The incidence of ARC was found to be directly related to the relative life expectancies in the breed groups: The smallest dog breeds had a lower ARC prevalence between ages 4 and 5 than mid-size breeds and these, in turn, a lower prevalence than the giant breeds. A similar sequence was evident for ages 7 to 10 and for overall lifetime incidence of ARC. These differences became more significant when comparing small and giant breeds only. We could also confirm the inverse relationship between body size and life expectancy in these same sets of dog breeds. Our results show that body size, life expectancy, and ARC incidence are interrelated in dogs. Given that ARC has been shown to be at least partially caused by oxidative damage to lens epithelial cells and the internal lens, we suggest that it can be considered not only as a general biomarker for life expectancy in the canine and possibly other species, but also for the systemic damages produced by reactive oxygen species. This suggests new approaches to examine the gene expression pathways affecting the above-noted linkages.

Inbreeding and fertility in Irish Wolfhounds in Sweden: 1976 to 2007.

BACKGROUND: Given that no influence of inbreeding on life expectancy could be demonstrated in Irish Wolfhounds in a previous study, it was decided to test the influence of inbreeding and other parameters on fertility in this breed. METHODS: The study was based on all Irish Wolfhound litters registered in Sweden between 1976 and 2007 (n = 822 litters) as provided by the Swedish Kennel Club (SKK) and combined with a pedigree database going back to 1862. Analyses were performed using linear regression in a Generalised Linear Model and other tests in the SAS system. RESULTS: Mean number of pups per litter was 6.01 +/- 2.65, with a maximum of 13. There were no significant differences in either the number of litters or the number of pups between years of birth. Males were used for breeding at a significantly earlier age than females. Mean number of litters per parent was 2.96 +/- 3.14 for males and 1.59 +/- 0.87 for females. No influence of Wright's inbreeding coefficients over 5, 10, 20 and 30 generations and/or Meuwissen's inbreeding coefficients on litter size was detected. In the Generalised Linear Model, highly significant, but weak (coefficient of determination (R2) = 0.0341) influences were found for maternal age at mating as well as maternal inbreeding measured by Wright's inbreeding coefficient over 30 generations and Meuwissen's inbreeding coefficient. Paternal inbreeding coefficients over 5, 10, 20 and 30 generations and calculated after Meuwissen, as well as maternal inbreeding coefficients over 5, 10 and 20 generations did not have significant effects on litter size. CONCLUSION: The low coefficient of determination (R2) value of the Generalised Linear Model indicates that inbreeding does not have a strong influence on fertility in Irish Wolfhounds, which is consistent with earlier results and the breed's genetic history. These results likely reflect the aforementioned genetic history and should not be extrapolated to other breeds without prior breed-specific research.