A Genetic Tool to Identify Predators Responsible for Livestock Attacks in South America and Recommendations for Human-Wildlife Conflict Mitigation.

Livestock predation induces global human-wildlife conflict, triggering the retaliatory killing of large carnivores. Although domestic dogs (Canis familiaris) contribute to livestock depredation, blame primarily falls on wild predators. Dogs can also transmit pathogens between wildlife, domestic animals, and humans. Therefore, the presence of free-ranging dogs can have negative consequences for biodiversity conservation, smallholder economy, food supply, and public health, four of the United Nations' Sustainable Developed Goals (SDGs) for 2030. In Ecuador, where livestock sustains rural households, retaliatory poaching threatens Andean bear (Tremarctos ornatus), jaguar (Panthera onca), and puma (Puma concolor) populations. However, the role of dogs in these incidents remains underexplored. The present study evaluates the possibility of reliable molecular identification of predatory species from DNA traces in bite wounds. Our results revealed the presence of dog saliva on four out of six livestock carcasses presumably attacked by wild predators. These findings highlight the importance of rectifying misinformation about large carnivores in Ecuador and the need to control dog populations. We recommend that local administrations incorporate DNA analysis into livestock predation events to examine how common the problem is, and to use the analysis to develop conflict mitigation strategies which are essential for the conservation of large carnivores.

Comparative Tensile Properties and Collagen Patterns in Domestic Cat (Felis catus) and Dog (Canis lupus familiaris) Ovarian Cortical Tissues.

The importance of the ovarian extracellular environment and tissue rigidity on follicle survival and development has gained attention in recent years. Our laboratory has anecdotally observed differences in the rigidity of domestic cat and dog ovarian cortical tissues, which have been postulated to underlie the differences in in vitro culture responses between the species, wherein cat ovarian tissues display higher survival in extended incubation. Here, the tensile strengths of cat and dog ovarian cortical tissues were compared via micropipette aspiration. The underlying collagen patterns, including fiber length, thickness, alignment, curvature, branch points and end points, and overall tissue lacunary and high-density matrix (HDM) were quantified via picrosirius red staining and TWOMBLI analysis. Finally, we explored the potential of MMP (-1 and -9) and TIMP1 supplementation in modulating tissue rigidity, collagen structure, and follicle activation in vitro. No differences in stiffness were observed between cat or dog cortical tissues, or pre- versus post-pubertal status. Cat ovarian collagen was characterized by an increased number of branch points, thinner fibers, and lower HDM compared with dog ovarian collagen, and cat tissues exposed to MMP9 in vitro displayed a reduced Young's modulus. Yet, MMP exposure had a minor impact on follicle development in vitro in either species. This study contributes to our growing understanding of the interactions among the physical properties of the ovarian microenvironment, collagen patterns, and follicle development in vitro.

First identification of Strongyloides stercoralis infection in a pet dog in Argentina, using integrated diagnostic approaches.

BACKGROUND: Strongyloides stercoralis is a soil-transmitted intestinal nematode with a complex life cycle that primarily affects humans, non-human primates, dogs, and occasionally cats. This study presents, to the best of our knowledge, the first case of S. stercoralis infection and its genotyping in a domestic dog from Argentina. METHODS: The patient was a female wired-haired Teckel dog exhibiting recurrent coughing. Coproparasitological analysis using the Baermann technique revealed the presence of rhabditiform larvae morphologically compatible with S. stercoralis. To confirm this finding, molecular diagnosis (18S ribosomal RNA) and analysis of the cox1 gene were performed. RESULTS: We identified a haplotype (HP20) that has previously only been related to S. stercoralis infection in dogs, but was found in the present study to be highly related to the haplotype (HP16) of a zoonotic variant and divergent from those previously described from human patients in Argentina. Furthermore, unlike in human cases following treatment with ivermectin, the dog was negative after moxidectin treatment according to polymerase chain reaction of the sampled faeces. CONCLUSIONS: This case report shows the importance of further investigation into potential transmission events and prevalences of S. stercoralis in dogs and humans in South America. The results reported here should also encourage future work that examines different scenarios of infection with S. stercoralis in dogs and humans with the aim of integrating clinical management, diagnosis, treatment and follow-up strategies in the quest for new approaches for the treatment of this disease in animals and humans. The findings support the adoption of a One Health approach, which recognizes the interconnectedness between animal and human health, in addressing parasitic infections such as strongyloidiasis.

Intraspecific Interactions Decrease Stress Affecting Welfare in Shelter Dogs: A Comparison of Four Different Housing Conditions.

Shelters are stressful environments for domestic dogs (Canis familiaris). Evaluating dogs' welfare is crucial to improve their life condition and to promote a better management of shelters. We aimed at verifying which variables improved welfare in 10 shelter dogs ((hosted in the shelter "Centro cinofilo Caerite" in Bracciano (Rome)) by analysing their behavioural responses in different environmental conditions. Furthermore, faecal samples were taken to measure cortisol metabolites (CM), a non-invasive method to evaluate adrenocortical activity in dogs. Dogs were observed for a total of 400 h in 4 different cage conditions: (i) alone in a cage; ii) alone in an enriched cage; (iii) in cage with conspecifics; (iv) in cage with regular interaction with humans outside the cage. Alone in the cage situation showed highest frequencies of displacement activities (Friedman test: χ2 = 13.32; p = 0.004). In contrast, being in the cage with conspecifics seems to reduce displacement activity frequency, as well as the level of faecal cortisol metabolites (Friedman test: χ2 = 8.04; p = 0.045). Our results suggest that conspecifics' presence is the best way to reduce stress in shelter dogs. This research could provide some useful guidelines for managing shelters and improving dogs' life condition.

Efficacy of Edible and Leisure Reinforcers with Domestic Dogs.

Preference assessments are often used to identify stimuli that function as potential reinforcers for training or intervention purposes. Specifically, various preference assessment formats have been used to identify preferred stimuli for humans, cockroaches, cotton-top tamarins, tortoises, and wolves, to name a few. However, to date, no study has evaluated the differential efficacy between food and leisure stimuli within domestic dogs. The current study aimed to compare the reinforcing value and efficacy between food and leisure stimuli for domestic dogs by comparing rates of behavior when receiving access to either their top-preferred food or leisure items. Overall results suggest (1) domestic dogs prefer food over leisure items, and (2) food is more likely to function as a reinforcer than leisure items for domestic dog's behavior. These results suggest that dog owners and trainers should consider using food reinforcers over leisure items as reinforcers when attempting to train dogs.

CanISO: a database of genomic and transcriptomic variations in domestic dog (Canis lupus familiaris).

BACKGROUND: The domestic dog, Canis lupus familiaris, is a companion animal for humans as well as an animal model in cancer research due to similar spontaneous occurrence of cancers as humans. Despite the social and biological importance of dogs, the catalogue of genomic variations and transcripts for dogs is relatively incomplete. RESULTS: We developed CanISO, a new database to hold a large collection of transcriptome profiles and genomic variations for domestic dogs. CanISO provides 87,692 novel transcript isoforms and 60,992 known isoforms from whole transcriptome sequencing of canine tumors (N = 157) and their matched normal tissues (N = 64). CanISO also provides genomic variation information for 210,444 unique germline single nucleotide polymorphisms (SNPs) from the whole exome sequencing of 183 dogs, with a query system that searches gene- and transcript-level information as well as covered SNPs. Transcriptome profiles can be compared with corresponding human transcript isoforms at a tissue level, or between sample groups to identify tumor-specific gene expression and alternative splicing patterns. CONCLUSIONS: CanISO is expected to increase understanding of the dog genome and transcriptome, as well as its functional associations with humans, such as shared/distinct mechanisms of cancer. CanISO is publicly available at https://www.kobic.re.kr/caniso/ .

Establishing a Sequencing Method for the Whole Mitochondrial DNA of Domestic Dogs.

In human beings, whole mitochondrial DNA (mtDNA) sequencing has been widely used in many research fields, including medicine, forensics, and genetics. With respect to the domestic dog (Canis lupus familiaris), which is commonly recognized as being an additional member of the traditional human family structure, research studies on mtDNA should be developed to expand and improve our collective knowledge of dog medicine and welfare as it seems that there is still room for further development in these areas. Moreover, a simple and robust method for sequencing whole mtDNA that can be applied to various dog breeds has not yet been described in the literature. In the present study, we aim to establish such a method for the whole mtDNA sequencing of the domestic dog. In the experiments we conducted, oral mucosa DNA samples obtained from six Japanese domestic dogs were used as a template. We designed four primer pairs that could amplify approximately 5 kbp from each region of the mtDNA and validated several PCR conditions. Subsequently, the PCR amplicons were pooled and subjected to library preparation. The sequencing of the libraries was performed using next-generation sequencing (NGS), followed by bioinformatics analysis. Our results demonstrate that the proposed method can be used to perform highly accurate resequencing. We believe that this method may be useful for future research conducted to better understand dog medicine and welfare.

Evolutionary history of the extinct wolf population from France in the context of global phylogeographic changes throughout the Holocene.

Phylogeographic patterns in large mammals result from natural environmental factors and anthropogenic effects, which in some cases include domestication. The grey wolf was once widely distributed across the Holarctic, but experienced phylogeographic shifts and demographic declines during the Holocene. In the 19th-20th centuries, the species became extirpated from large parts of Europe due to direct extermination and habitat loss. We reconstructed the evolutionary history of the extinct Western European wolves based on the mitogenomic composition of 78 samples from France (Neolithic-20th century) in the context of other populations of wolves and dogs worldwide. We found a close genetic similarity of French wolves from ancient, medieval and recent populations, which suggests the long-term continuity of maternal lineages. MtDNA haplotypes of the French wolves showed large diversity and fell into two main haplogroups of modern Holarctic wolves. Our worldwide phylogeographic analysis indicated that haplogroup W1, which includes wolves from Eurasia and North America, originated in Northern Siberia. Haplogroup W2, which includes only European wolves, originated in Europe ~35 kya and its frequency was reduced during the Holocene due to an expansion of haplogroup W1 from the east. Moreover, we found that dog haplogroup D, currently restricted to Europe and the Middle East, was nested within the wolf haplogroup W2. This suggests European origin of haplogroup D, probably as a result of an ancient introgression from European wolves. Our results highlight the dynamic evolutionary history of European wolves during the Holocene, with a partial lineage replacement and introgressive hybridization with local dog populations.

Inflammaging in domestic dogs: basal level concentrations of IL-6, IL-1ß, and TNF-α in serum of healthy dogs of different body sizes and ages.

Inflammaging, a "hallmark of aging," refers to a chronic, progressive increase in the proinflammatory status of mammals as they age, and this phenotype has been associated with many age-related diseases, including cardiovascular disease, arthritis and cancer. Though, inflammaging research is common in humans, there is a lack of data for this process for the domestic dog. Here, serum concentrations of IL-6, IL-1ß and TNF-α in healthy dogs of different body sizes and ages were measured to determine whether inflammaging may play a mechanistic role in aging rates in dogs, similar to those found in humans. Using a four-way ANOVA, a significant decrease in IL-6 concentrations in young dogs with the rest of the age categories showing increased IL-6 concentrations was found, similar to humans. However, only young dogs have decreased IL-6 concentrations, with adult dogs having similar IL-6 concentrations to senior and geriatric dogs, implying differences in aging rates between humans and dogs. And, there was a marginally significant interaction between sex*spayed or neutered status and IL-1ß concentrations with intact females having the lowest IL-1ß concentrations compared with intact males, and spayed and neutered dogs. The presence of estrogen in intact females may, overall, decrease inflammatory pathways. This implies that age at spaying or neutering may be an important aspect to consider for inflammaging pathways in dogs. Furthermore, sterilized dogs often die of immune-related diseases, which could be linked to the increases in IL-1ß in sterilized dogs found in this study.

Dog Life Spans and the Evolution of Aging.

AbstractThe basic tenets of the evolutionary theories of senescence are well supported. However, there has been little progress in determining the relative influences of mutation accumulation and life history optimization. The causes of the well-established inverse relationship between life span and body size across dog breeds are used here to test these two classes of theories. The life span-body size relationship is confirmed for the first time after controlling for breed phylogeny. The life span-body size relationship cannot be explained by evolutionary responses to differences in extrinsic mortality either of contemporary breeds or of breeds at their establishment. The development of breeds larger and smaller than ancestral gray wolves has occurred through changes in early growth rate. This may explain the increase in the minimum age-dependent mortality rate with breed body size and thus higher age-dependent mortality throughout adult life. The main cause of this mortality is cancer. These patterns are consistent with the optimization of life history as described by the disposable soma theory of the evolution of aging. The dog breed life span-body size relationship may be the result of the evolution of greater defense against cancer lagging behind the rapid increase in body size during recent breed establishment.

Molecular epidemiology of Canine circovirus in domestic dogs and wildlife in Namibia, Africa.

Canine circovirus (CanineCV) is a DNA virus affecting domestic dogs and other wild carnivore species. Despite the potential implications for dogs' health and wildlife conservation, data on CanineCV presence, epidemiology and genetic features from Africa is still poor. In the present study, biological specimens collected between 2020 and 2022 from a total of 32 jackals and 575 domestic dogs were tested for the presence of CanineCV DNA to evaluate its frequency. Furthermore, sequencing was conducted on positive samples to characterize the strains and compare them with publicly available sequences through phylogenetic analysis. A high CanineCV prevalence was observed both in jackals (43.75%; 95 CI: 28.17% - 60.67%) and domestic dogs (27.13%; 95 CI: 23.66% - 30.91%). All aside from one Namibian strain formed an independent clade, suggestive of extremely rare introduction events, followed by local persistence, circulation, and evolution. Remarkably, different recombination events were observed involving strains from both jackals and domestic dogs, which testify to the likely strain exchange between these populations. Distinctive amino acid residues were also observed in jackals. The limitations of the considered host populations however prevent a definitive conclusion on host adaptation, biological, and clinical features. Further studies should be performed to expand our current knowledge of the CanineCV disease scenario in Namibia, other African regions, and associated host species in Africa.

Live attenuated Leishmania infantum centrin deleted mutant (LiCen-/-) as a novel vaccine candidate: A field study on safety, immunogenicity, and efficacy against canine leishmaniasis.

This study was designed to evaluate the safety, immunogenicity, and efficacy of a single dose of L. infantum (LiCen-/-) live attenuated candidate vaccine against canine leishmaniasis (CanL). Eighteen healthy domestic dogs with no anti-Leishmania antibodies and negative leishmanin skin test (LST) were randomly inoculated intravenously with either L. infantum (LiCen-/-) vaccine candidate in 10 dogs or phosphate-buffered saline (PBS) in 8 dogs. The safety, immunogenicity, and efficacy rate of L. infantum (LiCen-/-) vaccine candidate against CanL were evaluated by different criteria, including clinical manifestations, injection-site lesion, hematology and biochemistry values, anti-Leishmania antibodies using direct agglutination test (DAT), delayed-type hypersensitivity (DTH) using LST, and CD4+ and CD8+ T-cells subsets, as well as by measuring interferon (IFN-γ), interleukin (IL-23), IL-17, and IL-10 cytokines. Spleen aspiration and detection of Leishmania parasite using parasitological examinations (microscopy and culture) were performed in both vaccinated and control groups. Two months after intervention, each dog was challenged intraperitoneally (IP) with wide type (WT) L. infantum. Two-month follow-up post vaccination showed no clinical signs and serious side effects associated with the vaccination. A significant increase was found in the expression of IL-17, CD4+, and CD8+ gene transcripts in PBMCs, as well as increased levels of Th1 cytokines, and reduction of Th2 cytokine. The efficacy of the vaccine candidate was calculated to be 42.85%. While the time window for assessing the vaccine's effectiveness was too limited to draw any real conclusions but the preliminary results showed a moderate efficacy rate due to inoculation a single dose of L. infantum (LiCen-/-) vaccine candidate. Further investigations with more sample sizes and multiple doses of the vaccine candidate using natural challenges in the endemic areas of CanL are recommended.

Social interaction can function as a reinforcer for dogs: Effects of stimulus duration and session parameters.

Prior research has produced mixed results on whether human social interaction can function as a reinforcer for dog behavior. However, that research used either short durations of social interaction or rapid, repeated trials such that satiation could have been a factor. We investigated whether two durations of social interaction (30 s or 4 s petting plus vocal praise) would maintain more responding than extinction, than each other, or than food. We limited each session to 10 trials and temporally spaced sessions within and across days. Both durations of social interaction produced more responding than extinction, but there was no difference in responding between the two social interaction durations. When we compared responding in food sessions to 30-s and 4-s social interaction sessions, we could not determine differences in responses emitted per session for two dogs due to ceiling effects, but the third dog doubled her responding when food was provided. Additionally, latencies in food sessions for all dogs were significantly lower than expected from a random sampling of latencies. Our results suggest both durations of social interaction can function as a reinforcer, especially when delivered sporadically, but they are still not as effective as food as a reinforcer for most dogs.

High Investment in Reproduction Is Associated with Reduced Life Span in Dogs.

AbstractProminent differences in aging among and within species present an evolutionary puzzle. The theories proposed to explain evolutionary differences in aging are based on the axiom that selection maximizes fitness, not necessarily life span. This implies trade-offs between investment in self-maintenance and investment in reproduction, where high investments in growth and current reproduction are associated with short life spans. Fast growth and large adult size are related to shorter life spans in the domestic dog, a bourgeoning model in aging research; however, whether reproduction influences life span in this system remains unknown. Here we test the relationship between reproduction and differences in life span among dog breeds, simultaneously controlling for shared ancestry and recent gene flow. We found that shared ancestry explains a higher proportion of the among-breed variation in life history traits, in comparison with recent gene flow. Our results also show that reproductive investment negatively impacts life span, and more strongly so in large breeds, an effect that is not merely a correlated response of adult size. These results suggest that basic life history trade-offs are apparent in a domestic animal whose diversity is the result of artificial selection and that among-breed differences in life span are due to a combination of size and reproduction.

Appeasement function of displacement behaviours? Dogs' behavioural displays exhibited towards threatening and neutral humans.

Appeasement signals are behavioural patterns displaying an animal's non-aggressive attitude and are hypothesized to reduce the aggressive behaviours in the receiver. In domestic dogs, specific displacement behaviours (i.e., behavioural patterns exhibited without an apparent function related to the ongoing situation), have been suggested to function as appeasement signals. To test this possibility, we assessed whether the occurrence of these behaviours was dependent on a social conflict context, predicting that, if displacement behaviours also function as appeasement signals, they should be more prevalent in a conflict vs. non-conflict context. Fifty-three dogs were exposed to two unfamiliar humans approaching them in either a mildly threatening or neutral way. We categorized the attitude of the dogs towards the strangers as "reactive", i.e., barking and lunging towards the stimulus, and "non-reactive", i.e., remaining passive in front of the stimuli. We coded dogs' displacement activities and modelled their duration or frequency as a function of the interaction between the test condition and the attitude of the dog. Displacement behaviours of "blinking", "nose licking" and "lip wiping" were associated with a "non-reactive" attitude, independently from the test condition, confirming an association with a non-aggressive intention. "Head turning" was associated with a "non-reactive" attitude in the threatening condition. In conclusion, dogs with a non-aggressive attitude exhibited more putative appeasement signals; however, these were not strictly associated with a conflict-ridden situation, calling for further investigation of their function.

Head-mounted mobile eye-tracking in the domestic dog: A new method.

Humans rely on dogs for countless tasks, ranging from companionship to highly specialized detection work. In their daily lives, dogs must navigate a human-built visual world, yet comparatively little is known about what dogs visually attend to as they move through their environment. Real-world eye-tracking, or head-mounted eye-tracking, allows participants to freely move through their environment, providing more naturalistic results about visual attention while interacting with objects and agents. In dogs, real-world eye-tracking has the potential to inform our understanding of cross-species cognitive abilities as well as working dog training; however, a robust and easily deployed head-mounted eye-tracking method for dogs has not previously been developed and tested. We present a novel method for real-world eye-tracking in dogs, using a simple head-mounted mobile apparatus mounted onto goggles designed for dogs. This new method, adapted from systems that are widely used in humans, allows for eye-tracking during more naturalistic behaviors, namely walking around and interacting with real-world stimuli, as well as reduced training time as compared to traditional stationary eye-tracking methods. We found that while completing a simple forced-choice treat finding task, dogs look primarily to the treat, and we demonstrated the accuracy of this method using alternative gaze-tracking methods. Additionally, eye-tracking revealed more fine-grained time course information and individual differences in looking patterns.

Comparison of behavioural tendencies between "dangerous dogs" and other domestic dog breeds - Evolutionary context and practical implications.

Aggressive behaviour by dogs is a considerable social problem, but the ability to predict which individuals may have increased aggressive tendencies is very limited, restricting the development of efficient preventive measures. There is a common perception that certain breeds are more likely to exhibit aggressive behaviour, which has contributed to the introduction of breed-specific legislation. The rationale for such legislation explicitly assumes high heritability of this trait while also implying relatively little variation within breeds; these assumptions are largely untested. We compared behavioural tendencies between 8 breeds that are subject to legislation in at least one country and 17 breeds that are not subject to legislation using two validated psychometric tools: the Dog Impulsivity Assessment Scale (DIAS), which scores elements of impulsivity, including a tendency for aggressive behaviour, and Positive and Negative Activation Scale (PANAS), which scores sensitivity to positive and negative stimuli (which may trigger aggressive responses). We found that the two groups of breeds do not differ significantly in the specific DIAS factor relating to aggressive behaviour, "Aggression Threshold and Response to Novelty", or any other DIAS and PANAS factors. We found large variations in all behavioural tendencies measured by both psychometric scales within both groups and within each breed studied. Taken together, our findings indicate that breed alone is not a reliable predictor of individual behavioural tendencies, including those related to aggression, and therefore breed-specific legislation is unlikely to be an effective instrument for reducing risk.

Characterization of the genetics and epidemiology of Brugia sp. in domestic dogs in Chad, Africa.

Neglected tropical diseases pose a threat to domestic animal health, as domestic animals can serve as reservoirs for certain zoonotic parasitic infections, including Guinea worm (Dracunculus medinensis) and lymphatic filariasis. Surveillance for these parasites in domestic animals is needed to understand infection prevalence and transmission cycles, with the goal of instituting appropriate interventions. The goal of this research was to report our finding of Brugia sp. infection in dogs from Chad, Africa, and to characterize the genetics and epidemiology of the parasite. During a recent Chadian canine pathogen surveillance project, we identified Brugia sp. infections in a total of 46 out of 428 dogs (10.7%) sampled at three time points in 2019-2020. We found high levels of sequence similarity to B. malayi and B. pahangi based on amplification of 18S rRNA, 5.8S rRNA, and ITS-2 regions. Phylogenetic analysis of 18S rRNA gene sequences placed the Chadian Brugia sp. in a clade with other Brugia spp. but grouped it separately from both B. malayi and B. pahangi. Analysis of Hha I sequences showed the greatest similarity with B. patei, a parasite previously reported from dogs, cats, and wildlife hosts in Kenya. Epidemiologic analysis using generalized linear regression modeling found significantly higher odds of Brugia sp. detection among dogs in villages in southern Chad compared to those in the northern region. Further, within the northern region, there were higher odds of detection in the dry season, compared to the wet season, which is consistent with the ecology of a presumably mosquito-borne parasite. The same 428 dogs were tested for Dirofilaria immitis antigen using a commercial assay (IDEXX SNAP 4Dx) at the earliest time point of the study, with 119 dogs testing positive. However, no association was noted between Brugia infection and a dog being positive for Di. immitis antigen, with only seven of the 119 Di. immitis antigen-positive dogs being Brugia-positive. This is the first report of Brugia sp. in domestic dogs in Chad and additional research is needed to definitively identify the species present, elucidate transmission, and understand potential risks to canine and human health.

Interspecific coprophagia by wild red foxes: DNA metabarcoding reveals a potentially widespread form of commensalism among animals.

Vertebrate animals are known to consume other species' faeces, yet the role of such coprophagy in species dynamics remains unknown, not least due to the methodological challenges of documenting it. In a large-scale metabarcoding study of red fox and pine marten scats, we document a high occurrence of domestic dog DNA in red fox scats and investigate if it can be attributed to interspecific coprophagia. We tested whether experimental artifacts or other sources of DNA could account for dog DNA, regressed dog occurrence in the diet of fox against that of the fox's main prey, short-tailed field voles, and consider whether predation or scavenging could explain the presence of dog DNA. Additionally, we determined the calorific value of dog faeces through calorimetric explosion. The high occurrence of dog DNA in the diet of fox, the timing of its increase, and the negative relationship between dog and the fox's main prey, point to dog faeces as the source of DNA in fox scats. Dog faeces being highly calorific, we found that foxes, but not pine martens, regularly exploit them, seemingly as an alternative resource to fluctuating prey. Scattered accounts from the literature may suggest that interspecific coprophagia is a potentially frequent and widespread form of interaction among vertebrates. However, further work should address its prevalence in other systems and the implications for ecological communities. Tools such as metabarcoding offer a way forward.

Pet dogs' Behavioural Reaction to Their Caregiver's Interactions with a Third Party: Join in or Interrupt?

Pet dogs are promising candidates to study attachment-related and potentially jealousy-like behaviours in non-human animals, as they form a strong and stable bond with their human caregivers who often engage in affiliative interactions with diverse social partners. Nevertheless, it is still debated whether non-human animals are capable of experiencing such complex emotions. Even though caregivers frequently report observations of jealousy-like behaviours in dogs, behavioural studies in dogs have thus far led to contradictory results. Adding to this complexity, dogs appear extraordinarily skilled in understanding humans' communicative behaviour and can flexibly and diversely interact with them in social contexts. Here, we aimed at investigating (1) whether dogs indeed respond in a jealousy-consistent manner when seeing their caregiver interact in an affiliative way with a remotely controlled, realistic-looking fake dog, or (2) whether they would rather synchronize their reaction to the fake dog with the caregiver's behaviour, or (3) whether they respond directly to the caregiver without paying much attention to the third party. To address what drives the dogs' behaviours in this triadic situation, we compared four groups of dogs who first observed and then joined the interaction of either the caregiver or a stranger greeting or medically examining the fake dog. We found that the dogs initially responded negatively or neutrally when the fake dog entered the room but changed to more positive reactions when the caregiver approached the fake dog, especially if initiating a positive interaction. When being released, more dogs showed friendly behaviours towards the fake dog when the caregiver-rather than the stranger-was interacting with it. At the same time, however, the dogs tried to block the interaction of the caregiver with the fake dog more often than the one of the stranger. In conclusion, we did not find clear evidence for jealousy-like behaviours in dogs during the human-fake dog interactions, but we observed indicators of behavioural synchronization with the caregivers, suggesting that the caregivers' affiliative behaviours directed at a third party may more often facilitate positive than negative interactions in dogs.