Human Animal Research Articles

Encapsulation of Saccharomyces spp. for Use as Probiotic in Food and Feed: Systematic Review and Meta-analysis.

Probiotics, particularly yeasts from the genus Saccharomyces, are valuable for their health benefits and potential as antibiotic alternatives. To be effective, these microorganisms must withstand harsh environmental conditions, necessitating advanced protective technologies such as encapsulation to maintain probiotic viability during processing, storage, and passage through the digestive system. This review and meta-analysis aims to describe and compare methods and agents used for encapsulating Saccharomyces spp., examining operating conditions, yeast origins, and species. It provides an overview of the literature on the health benefits of nutritional yeast consumption. A bibliographic survey was conducted following the Preferred Reporting Items for Systematic Review and Meta-Analysis (PRISMA) guidelines. The meta-analysis compared encapsulation methods regarding their viability after encapsulation and exposure to the gastrointestinal tract. Nineteen studies were selected after applying inclusion/exclusion criteria. Freeze drying was found to be the most efficient for cell survival, while ionic gelation was best for maintaining viability after exposure to the gastrointestinal tract. Consequently, the combination of freeze drying and ionic gelation proved most effective in maintaining high cell viability during encapsulation, storage, and consumption. Research on probiotics for human food and animal feed indicates that combining freeze drying and ionic gelation effectively protects Saccharomyces spp.; however, industrial scalability must be considered. Reports on yeast encapsulation using agro-industrial residues as encapsulants offer promising strategies for preserving potential probiotic yeasts, contributing to the environmental sustainability of industrial processes.

Iron-depleting nutritional immunity controls extracellular bacterial replication in Legionella pneumophila infections

The accidental human pathogen Legionella pneumophila (Lp) is the etiological agent for a severe atypical pneumonia known as Legionnaires’ disease. In human infections and animal models of disease alveolar macrophages are the primary cellular niche that supports bacterial replication within a unique intracellular membrane-bound organelle. The Dot/Icm apparatus—a type IV secretion system that translocates ~300 bacterial proteins within the cytosol of the infected cell—is a central virulence factor required for intracellular growth. Mutant strains lacking functional Dot/Icm apparatus are transported to and degraded within the lysosomes of infected macrophages. The early foundational work from Dr. Horwitz’s group unequivocally established that Legionella does not replicate extracellularly during infection—a phenomenon well supported by experimental evidence for four decades. Our data challenges this paradigm by demonstrating that macrophages and monocytes provide the necessary nutrients and support robust Legionella extracellular replication. We show that the previously reported lack of Lp extracellular replication is not a bacteria intrinsic feature but rather a result of robust restriction by serum-derived nutritional immunity factors. Specifically, the host iron-sequestering protein Transferrin is identified here as a critical suppressor of Lp extracellular replication in an iron-dependent manner. In iron-overload conditions or in the absence of Transferrin, Lp bypasses growth restriction by IFNγ-primed macrophages though extracellular replication. It is well established that certain risk factors associated with development of Legionnaires’ disease, such as smoking, produce a chronic pulmonary environment of iron-overload. Our work indicates that iron-overload could be an important determinant of severe infection by allowing Lp to overcome nutritional immunity and replicate extracellularly, which in turn would circumvent intracellular cell intrinsic host defenses. Thus, we provide evidence for nutritional immunity as a key underappreciated host defense mechanism in Legionella pathogenesis.

Preventing future zoonosis: SARS-CoV-2 mutations enhance human–animal cross-transmission

The COVID-19 pandemic has driven substantial evolution of the SARS-CoV-2 virus, yielding subvariants that exhibit enhanced infectiousness in humans. However, this adaptive advantage may not universally extend to zoonotic transmission. In this work, we hypothesize that viral adaptations favoring animal hosts do not necessarily correlate with increased human infectivity. In addition, we consider the potential for gain-of-function mutations that could facilitate the virus’s rapid evolution in humans following adaptation in animal hosts. Specifically, we identify the SARS-CoV-2 receptor-binding domain (RBD) mutations that enhance human–animal cross-transmission. To this end, we construct a multitask deep learning model, MT-TopLap trained on multiple deep mutational scanning datasets, to accurately predict the binding free energy changes upon mutation for the RBD to ACE2 of various species, including humans, cats, bats, deer, and hamsters. By analyzing these changes, we identified key RBD mutations such as Q498H in SARS-CoV-2 and R493K in the BA.2 variant that are likely to increase the potential for human–animal cross-transmission.

The Welfare of Dogs and Cats in the European Union: A Gap Analysis of the Current Legal Framework.

Companion animals, especially dogs and cats, have increasingly been recognized as moral subjects and valued as family members by European citizens. This new role encourages policy makers to face the many companion animals' welfare issues not yet covered by the EU legislation. The main gaps in the protection of dog and cat welfare during their all lifespan have been identified: unhealthy and unethical breeding practices, irresponsible sale and ownership, transport for commercial and non-commercial purposes, training methods, working dog conditions and rights, regulation of dog and cat shows and competitions, the therapeutic neglect, dog and cat end-of-life care, shelter management legislation and the free-ranging cat and dog population rights. The EU legislation framework is still very weak and far from establishing a harmonized approach, promoting a high standard of care and protection across Member States. We conclude that education and awareness regarding responsible pet ownership and the need for a One Welfare approach have a high value in finding adequate solutions, especially when poor human social welfare affects companion animal welfare. Given the link between human and companion animal welfare, the use of stakeholders' involvement strategies and a transdisciplinary approach appear crucial for the development of an EU legal framework for the well-being of dogs and cats.

Human animal relationships in Bos indicus cattle breeds addressed from a Five Domains welfare framework.

The present review has two objectives, the first is to investigate the differences in temperament between Bos indicus and Bos taurus breeds and determining the effects on production due to positive treatment and to compare this with negative HAR, by using the Five Domain Model as framework. The second objective is to discuss potential strategies to achieve better HAR when working with Bos indicus cattle. Bos indicus are more reactive and temperamental than Bos taurus cattle. When human animal relationships (HAR) are evaluated, Bos indicus cattle may react with greater intensity. They may be more likely to develop a negative emotional state, especially in extensively raised Bos indicus cattle that are handled only a few times each year. Bos indicus cattle can have positive emotional states when they have frequent positive interactions with people. Interactions with people, both positive and negative, would be in the fourth Domain of the Five Domains model. Cattle that are more reactive during handling may also have lower weight gain, even when they have abundant feed. This would be in the first Domain of Nutrition. When cattle are handled in races and corrals, injuries may be more likely to occur. Injuries and bruises would be in the third Domain of Health. Injuries could be caused by either poor handling practices by people or poor handling facilities. Yelling or electric prod use would be examples of poor HAR. Second Environmental Domain issues may be broken facilities or slick, slippery floors that are associated with falls.

The culture of care to enhance laboratory animal personnel well-being: a scoping review.

There has been intense focus on improving the quality of animal research in recent times. An emerging concept of a 'culture of care' has been proposed as another important pillar to enhance scientific quality, with staff well-being being a critical aspect. Professionals working with research animals can face moral and psychological burdens and are at risk of experiencing work-related stress. However, data on the global prevalence of stress in this population is limited. Equally, it is not clear how these stresses manifest, and what impact they might have on an individual's workplace performance and research quality. The purpose of this review was to identify work-related stress, its prevalence, and map evidence on strategies to mitigate stresses. We also set out to identify studies assessing the association between work-related stress and research quality. A systematic search was conducted across four databases, in addition to hand searching relevant references. We included peer-reviewed publications describing work-related stress, culture of care and laboratory animal professionals. A total of 49 publications were included for data mapping. Compassion fatigue was the most frequently described work-related stress, and its prevalence across Europe and North America is likely to be widespread. Multiple strategies to mitigate compassion fatigue and work-related stress were put forward, however, limited empirical evidence was available to assess success. Moreover, no studies reported empirical data linking work-related stress with research quality, despite several publications stating the case. Further population-specific research and measured assessments are urgently needed to deliver culture of care programmes to improve human well-being, animal welfare and research quality.

Valproic acid-induced teratogenicity is driven by senescence and prevented by Rapamycin in human spinal cord and animal models.

Valproic acid (VPA) is an effective and widely used anti-seizure medication but is teratogenic when used during pregnancy, affecting brain and spinal cord development for reasons that remain largely unclear. Here we designed a genetic recombinase-based SOX10 reporter system in human pluripotent stem cells that enables tracking and lineage tracing of Neural Crest cells (NCCs) in a human organoid model of the developing neural tube. We found that VPA induces extensive cellular senescence and promotes mesenchymal differentiation of human NCCs. We next show that the clinically approved drug Rapamycin inhibits senescence and restores aberrant NCC differentiation trajectory after VPA exposure in human organoids and in developing zebrafish, highlighting the therapeutic promise of this approach. Finally, we identify the pioneer factor AP1 as a key element of this process. Collectively our data reveal cellular senescence as a central driver of VPA-associated neurodevelopmental teratogenicity and identifies a new pharmacological strategy for prevention. These results exemplify the power of genetically modified human stem cell-derived organoid models for drug discovery.

Rapid assessment of human–elephant conflict: a crime science approach

An interdisciplinary approach has the potential not only to help solve conservation-centric problems but also to enrich and improve evidence-based scientific research. Crime science, an offshoot of criminology, provides a comprehensive, solution-oriented approach that transcends disciplinary boundaries and bridges science and practice for developing effective conservation interventions to real-life problems such as Human Elephant Conflict (HEC). This paper focuses on HEC as a conservation concern, but the resultant behaviors toward elephants, people, and their property are criminology’s concern. Using the Action Research paradigm, a rapid assessment of human–elephant conflict (HEC) in India was undertaken to identify contextual solutions. This study utilized problem-oriented field research methods that enabled the gathering of data on elephant habitat-landscape, villagers’ lifestyle (habitat) in the fringe areas, their current approaches in dealing with the conflict, the challenges forest officials face to mitigate HEC, and the assistance provided by district administrators to protect villagers and their corps and HEC-related deaths. The qualitative inquiry, including observation of village-forest fringe areas, focus group discussions with villagers, and interviews with forest officers and rangers, and district administrators/collectors who are handlers, guardians, and managers of the conflict space, provided rich data in identifying situational practical measures and underscored the role of crime science in providing a conceptual framework to gather evidence in addressing HEC in forest areas. The findings of the research suggest that human–animal convergence space is the source (or location) of conflict and criminology-driven situational crime prevention measures, including increasing effort, risks, reducing rewards and provocations, and removing excuses might mitigate the conflict, requiring coordinated efforts by villagers, forest and district administrators, and local law enforcers.

A Cross-Cultural Comparison of Cat–Human Relationships in the United States and Japan

ABSTRACT Across the world, more and more cats are being kept in captive spaces such as homes, animal shelters, and cat cafés. There is a growing need to understand whether cross-cultural differences influence human–animal interactions in these human-controlled environments. Cats are one of the world’s most popular companion animals, and both the US and Japan have high rates of cat ownership. However, cultural differences may account for differences in how people feel about cats and how humans and cats behave toward one another. The research aim was to examine whether differences exist between the US and Japan across multiple facets of the human–cat relationship. Study 1 examined each country’s attitude toward pet cats. Results indicate US citizens have significantly more positive attitudes toward pet cats than Japanese citizens. Study 2 examined owner attachment to their cat and pet cat social behavior. Results indicate owners in both countries were highly attached to their cats. In the US, but not Japan, owner attachment level predicted pet cat sociability toward the owner. Study 3 examined cat social behavior in cat cafés. The results of studies 2 and 3 indicate that pet cats and café cats in the US spend a greater proportion of time in proximity to people compared with pet and café cats in Japan. In all, differences and similarities were seen between the US and Japan. In humans, differences in childhood cat experiences and lower rates of cat ownership may impact attitudes toward cats. In pet cats, owner characteristics, the size of the cat’s environment, cat care practices, and attitudes toward cats may impact cat social behavior. Future exploration in this area can benefit our understanding of cross-cultural human–animal interactions and how to apply these findings to improve captive-cat welfare.

A toxicogenomics-based identification of potential mechanisms and signaling pathways involved in PFCs-induced cancer in human.

The number of new diagnosed cancer cases and cancer deaths are increasing worldwide. Perfluorinated compounds (PFCs) are synthetic chemicals, which are possible inducers of cancer in human and laboratory animals. Studies showed that PFCs induce breast, prostate, kidney, liver and pancreas cancer by inducing genes being involved in carcinogenic pathways. This study reviews the association between PFCs induced up-regulation/down-regulation of genes and signaling pathways that are important in promoting different types of cancer. To obtain chemical-gene interactions, an advanced search was performed in the Comparative Toxicogenomics Database platform. Five most prevalent cancers were studied and the maps of their signaling pathways were drawn, and colored borders indicate significantly differentially expressed genes if there had been reports of alterations in expression in the presence of PFCs. In general, PFCs are capable of inducing cancer in human via altering PPARα and PI3K pathways, evading apoptosis, inducing sustained angiogenesis, alterations in proliferation and blocking differentiation. However, more epidemiological data and mechanistic studies are needed to better understand the carcinogenic effects of PFCs in human.

Nano-Rutin: A Promising Solution for Alleviating Various Disorders.

Rutin, often known as vitamin P, is a natural flavonoid compound, which offers a broad spectrum of therapeutic potentials. Rutin is metabolised to different compounds by the gut bacteria after consumption, therefore, very little is absorbed. Higher plants contribute to rutin synthesis in large quantities, and it may also be found in many fruits and fruity rinds, particularly citrus fruits and berries. The present paper highlights several studies conducted on rutin along with its nanoformulations regarding its broad spectrum of therapeutic potentials. Numerous electronic databases, including Springer, PubMed, Science Direct, Pubchem, etc. were searched to extract relevant published literature demonstrating rutin effectiveness in various ailments. The reviewed literature showed that rutin and related flavonoids possess a variety of physiological properties that protects human beings, plants and animals. Antioxidant, anti-inflammatory, anti-allergic, cytoprotective, vasoprotective, anticarcinogenic, neuroprotective, cardioprotective, antibacterial, antiviral, antiprotozoal, antitumor, anti-hypertensive antiplatelet, antispasmodic and hypolipidemic, activities. Nanotechnology has been implemented for the improvement of the bioavailability of rutin using novel drug-delivery carriers. The study concludes that the development of rutin nanoformulations for multiple therapeutic approaches contributes towards enhanced aqueous solubility as well as tailored pharmacokinetics compared to conventional delivery of rutin. However, more investigations should be conducted to confirm the improved bioavailability of the rutin nanoformulations.

Estimates of the burden of human rabies deaths and animal bites in India, 2022–23: a community-based cross-sectional survey and probability decision-tree modelling study

Reliable and recent data of human rabies deaths and animal bites are not available in India, where a third of global cases occur. Since there is a global target of eliminating dog-mediated human rabies by 2030, understanding whether the country is on track is essential. We aimed to estimate the animal-bite burden and the number of human rabies deaths in India. We conducted a community-based nationwide cross-sectional survey with a multistage cluster-sampling design from March 2, 2022 to Aug 26, 2023, covering 60 districts in 15 Indian states. The head of the household or an adult family member was interviewed to collect information about animal-bite history in family members, receipt of anti-rabies vaccination (ARV), and death following animal bite in the family. Annual animal-bite incidence along with 95% CIs were estimated after applying the sampling weights and adjusting for clustering. We estimated annual human rabies deaths using a decision-tree probability model with parameters from the community survey and laboratory data on rabies positivity among suspected rabid dogs. Of the 337 808 individuals residing in the 78 807 households surveyed, 2052 gave a history of animal bite, mostly (1576 [76·8%]) due to dogs in the past 1 year. The weighted and adjusted annual incidence of animal bite was 6·6 (95% CI 5·7-7·6) per 1000 population, translating into 9·1 million bites nationally. Annual dog-bite incidence was 5·6 (4·8-6·6) per 1000. Among people who had been bitten by a dog, 323 (20·5%) did not receive ARV, and 1043 (66·2%) received at least three doses. Nearly half (615 [49·1%]) of the 1253 individuals who received one dose did not complete their full course of vaccination. We estimated 5726 (95% uncertainty interval 3967-7350) human rabies deaths occurring annually in India. Although there was a substantial decline in human rabies deaths over the past two decades, to eliminate dog-mediated human rabies by 2030, India needs to fast-track its actions by adopting a focused one-health approach. Integrating human and animal surveillance, ensuring timely administration of full course of post-exposure prophylaxis, and accelerating dog vaccination across the country are crucial steps towards this goal. Indian Council of Medical Research.

A Suite of Pathogens Detected in Ticks Sampled From Wildlife Hosts in Central Kenya

ABSTRACTOpportunistic sampling of ticks from animals during veterinary treatment offers important insights about wildlife exposure to pathogens. Here, we report pathogens and other tick‐borne organisms detected in ticks sampled from 10 animal species in and near Laikipia County, Kenya, in 2014–2016. Pathogen analysis was performed using a combination of Fluidigm PCR and Illumina sequencing. We detected a suite of important pathogens affecting human and domestic and wild animal health, including Anaplasma marginale, Rickettsia africae and Theileria parva. These findings expand our limited understanding of wildlife exposure to tick‐borne pathogens and potential ramifications for animal health and conservation.

Development of a Simple and Rapid DNA Extraction Method for Aspergillus flavus

Abstract Aspergillus species are known to be very important in human and domestic animal health. Aspergillus species commonly cause severe systemic and skin infections, as well as allergic lung diseases. With the development of PCR techniques, these methods are used to identify and diagnose fungi. DNA extraction from Aspergillus species is difficult because the fungal cell wall structure is very durable and complex. Fungal DNA extraction methods containing proteinase K and liquid nitrogen are widely used to break down the cell wall. However, these methods cause DNA loss during the extraction in Aspergillus species. In this study, on the contrary, the commonly used DNA extraction by means of ammonium hydroxide, which is generally used to break down chitin in DNA extraction of ticks and plants, is used. The efficiency of the cell wall lysis method from A. flavus with ammonium hydroxide was compared with methods containing proteinase K and liquid nitrogen. For this purpose, DNA extraction of A. flavus was tried using three different methods. As a result, the cell wall of A. flavus was lysed using ammonium hydroxide in this study. The obtained DNA’s quality, concentration, and PCR performance were sufficient. This method has been evaluated as a faster, more straightforward, and more economical alternative.

P02-33 Bridging the gap: human and preclinical animal microphysiological systems for assessing drug-induced liver injury during drug discovery

P02-33 Bridging the gap: human and preclinical animal microphysiological systems for assessing drug-induced liver injury during drug discovery

The Chinese Freshwater Jellyfish Unbound

Jellyfish are commonly known as dangerous sea creatures that humans should avoid. Focusing on the newly discovered freshwater jellyfish in the late nineteenth century, this article explores a global history of Craspedacusta sowerbii, a species native to China first noted in nineteenth-century Britain but since traveling the world over. The worldwide distribution of C. sowerbii, as both polyps and medusae, made it a global subject of scientific inquiry. However, no independent study has examined the history behind the origin, circulation, and implication of the freshwater jellyfish and its relationship with marine types. The late nineteenth century was the period when our premodern knowledge of jellyfish transformed from it being purely a marine life to the modern recognition of jellyfish as both marine and freshwater organisms, triggered by the discovery and discussion of the Chinese freshwater jellyfish C. sowerbii among naturalists and philosophers. This article focuses on Arthur de Carle Sowerby’s engagement with C. sowerbii in terms of narratives of evolution, nomenclature, and bioinvasion between 1880 and 1941. I argue that jellyfish, as a lowly form of invertebrates, should be taken more seriously by historians of modern China and historians of biology who are interested in the diverse roles of animals in shaping human–animal interactions in the Anthropocene. While the scientific research of jellyfish’s global bloom abounds, the interpretation of the global rise of jellyfish could benefit from a historically and culturally bound analysis of this classical animal. This is particularly so as scientists recognized the diverse perceptions of jellyfish. The global jellyfish bloom requires historical perspectives on a topic that has hitherto received scant attention from historians of science.

Does wildlife-vehicle collision frequency increase on full moon nights? A case-crossover analysis

Wildlife-vehicle collisions (WVCs) raise concerns for both human safety and animal welfare. As the literature has reported increased animal-related crash frequency on full moon nights in several regions, we investigated if a similar pattern is observed in Texas. We counted WVC and non-WVC frequencies on full moon nights and new moon nights in Texas between January 2011 and January 2020. The analysis revealed a 45.80% (95% confidence interval (CI): 29.94%–61.29%) increase in WVCs on full moon nights compared to new moon nights, with no statistically significant difference for non-WVCs (95% CI: -2.58%–1.45%). The association was more pronounced in rural areas than in urban areas. It is likely that brighter moonlight is strongly associated with higher WVC rates. The results illuminate the importance of heightened caution for drivers even on bright nights, particularly when driving through areas with high wildlife density.

Beyond the incubator: applying a "one health" approach in the NICU.

A "one health" approach recognises that human health, animal health and planetary health are closely interlinked and that a transdisciplinary approach is required to fully understand and maintain global health. While, by necessity, Neonatal Intensive Care has traditionally focused on the acutely unwell newborn, the avoidance of long-term harm is core to many management decisions. The COVID 19 pandemic and climate crisis have brought into sharp relief the importance of a "one health" approach as part of long-term health promotion in the holistic care of neonates, who may survive to experience the burden of future environmental crises. This narrative review seeks to integrate what we know about "one health" issues in the neonatal intensive care unit, notably antimicrobial resistance and climate change, and suggest "everyday changes" which can be utilised by practitioners to minimise the impact of neonatal intensive care on these global health issues. Many of the changes suggested not only represent important improvements for planetary health but are also core to good neonatal practice. IMPACT: Neonatal patients are likely to bear the burden of future environmental crises including pandemics and climate related disasters. While the focus of intensive care practitioners is acute illness, awareness of "one health" problems are important for our smallest patients as part of preventing long-term harm. High quality neonatal care can benefit both the planet and our patients.

CYP8B1 Catalyzes 12alpha-Hydroxylation of C27 Bile Acid: In Vitro Conversion of Dihydroxycoprostanic Acid into Trihydroxycoprostanic Acid.

Sterol 12α-hydroxylase (CYP8B1) is the unique P450 enzyme with sterol 12-oxidation activity, playing an exclusive role in 12α-hydroxylating intermediates along the bile acid (BA) synthesis pathway. Despite the long history of BA metabolism studies, it is unclear whether CYP8B1 catalyzes 12α-hydroxylation of C27 BAs, the key intermediates shuttling between mitochondria and peroxisomes. This work provides robust in vitro evidence that both microsomal and recombinant CYP8B1 enzymes catalyze the 12α-hydroxylation of dihydroxycoprostanic acid (DHCA) into trihydroxycoprostanic acid (THCA). On the one hand, DHCA 12α-hydroxylation reactivity is conservatively detected in liver microsomes of both human and preclinical animals. The reactivity of human tissue fractions conforms well with the selectivity of CYP8B1 mRNA expression, while the contribution of P450 enzymes other than CYP8B1 is excluded by reaction phenotyping in commercial recombinant enzymes. On the other hand, we prepared functional recombinant human CYP8B1 proteins according to a recently published protocol. Titration of the purified CYP8B1 proteins with either C4 (7α-hydroxy-4-cholesten-3-one) or DHCA yields expected blue shifts of the heme Soret peak (type I binding). The recombinant CYP8B1 proteins efficiently catalyze 12α-hydroxylation of both DHCA and C4, with substrate concentration occupying half of the binding sites of 3.0 and 1.9 μM and kcat of 3.2 and 2.6 minutes-1, respectively. In summary, the confirmed role of CYP8B1 in 12α-hydroxylation of C27 BAs has furnished the forgotten passageway in the BA synthesis pathway. The present finding might have opened a new window to consider the biology of CYP8B1 in glucolipid metabolism and to evaluate CYP8B1 inhibition as a therapeutic approach of crucial interest for metabolic diseases. SIGNIFICANCE STATEMENT: The academic community has spent approximately 90 years interpreting the synthesis of bile acids. However, the 12α-hydroxylation of intermediates catalyzed by CYP8B1 is not completely mapped on the classic pathway, particularly for the C27 bile acids, the pivotal intermediates shuttling between mitochondria and peroxisomes. This work discloses the forgotten 12α-hydroxylation pathway from dihydroxycoprostanic acid into trihydroxycoprostanic acid. The present finding may facilitate evaluating CYP8B1 inhibition as a therapeutic approach of crucial interest for metabolic diseases.

Exploring Levels of Interspecies Interaction: Expectations, Knowledge, and Empathy in Human-Dog Relationships.

This exploratory study examines the complex dynamics of human-dog relationships and their impact on interspecies communication. Twelve human-dog dyads were studied using narrative interviews to explore how people perceive their relationships with their dogs. In addition, the dyads engaged in a cooperative task to observe interaction dynamics during everyday activities. This study shows that individual expectations frame interactions and that traditional notions of dog ownership are evolving into more family-like relationships. Effective communication relies on a nuanced mix of verbal and non-verbal cues, with empathy emerging as a fundamental element guiding these interactions. Our findings underline the profound influence of human expectations, knowledge and empathy on communication with dogs. They also highlight the critical role of compatibility between human and dog dyads, and emphasize that such compatibility is a key determinant of satisfaction in interspecies relationships. These findings contribute to a deeper understanding of how human factors modulate communication and satisfaction in human-animal interactions.