Domestication constrains the ability of dogs to convey emotions via facial expressions in comparison to their wolf ancestors.

Dogs (Canis lupus familiaris) are the domestically bred descendant of wolves (Canis lupus). However, selective breeding has profoundly altered facial morphologies of dogs compared to their wolf ancestors. We demonstrate that these morphological differences limit the abilities of dogs to successfully produce the same affective facial expressions as wolves. We decoded facial movements of captive wolves during social interactions involving nine separate affective states. We used linear discriminant analyses to predict affective states based on combinations of facial movements. The resulting confusion matrix demonstrates that specific combinations of facial movements predict nine distinct affective states in wolves; the first assessment of this many affective facial expressions in wolves. However, comparative analyses with kennelled rescue dogs revealed reduced ability to predict affective states. Critically, there was a very low predictive power for specific affective states, with confusion occurring between negative and positive states, such as Friendly and Fear. We show that the varying facial morphologies of dogs (specifically non-wolf-like morphologies) limit their ability to produce the same range of affective facial expressions as wolves. Confusion among positive and negative states could be detrimental to human-dog interactions, although our analyses also suggest dogs likely use vocalisations to compensate for limitations in facial communication.

Behavioural synchronisation between different groups of dogs and wolves and their owners/handlers: Exploring the effect of breed and human interaction.

Dogs have previously been shown to synchronise their behaviour with their owner and the aim of this study was to test the effect of immediate interactions, breed, and the effects of domestication. The behavioural synchronisation test was conducted in outdoor enclosures and consisted of 30 s where the owner/handler was walking and 30 s of standing still. Three studies were conducted to explore the effect of immediate interaction (study A), the effect of breed group (study B), and the effect of domestication (study C). In study A, a group of twenty companion dogs of various breeds were tested after three different human interaction treatments: Ignore, Pet, and Play. The results showed that dogs adjusted their movement pattern to align with their owner's actions regardless of treatment. Furthermore, exploration, eye contact, and movement were all influenced by the owners moving pattern, and exploration also decreased after the Play treatment. In study B, the synchronisation test was performed after the Ignore treatment on three groups: 24 dogs of ancient dog breeds, 17 solitary hunting dogs, and 20 companion dogs (data from study A). Irrespective of the group, all dogs synchronised their moving behaviour with their owner. In addition, human walking positively influenced eye contact behaviour while simultaneously decreasing exploration behaviour. In study C, a group of six socialised pack-living wolves and six similarly socialised pack-living dogs were tested after the Ignore treatment. Interestingly, these animals did not alter their moving behaviour in response to their handler. In conclusion, dogs living together with humans synchronise with their owner's moving behaviour, while wolves and dogs living in packs do not. Hence, the degree of interspecies behavioural synchronisation may be influenced by the extent to which the dogs are immersed in everyday life with humans.

Fear of the human 'super predator' in native marsupials and introduced deer in Australia.

Recent experiments have demonstrated that carnivores and ungulates in Africa, Asia, Europe and North America fear the human 'super predator' far more than other predators. Australian mammals have been a focus of research on predator naiveté because it is suspected they show atypical antipredator responses. To experimentally test if mammals in Australia also most fear humans, we quantified the responses of four native marsupials (eastern grey kangaroo, Bennett's wallaby, Tasmanian pademelon, common brushtail possum) and introduced fallow deer to playbacks of predator (human, dog, Tasmanian devil, wolf) or non-predator control (sheep) vocalizations. Native marsupials most feared the human 'super predator', fleeing humans 2.4 times more often than the next most frightening predator (dogs), and being most, and significantly, vigilant to humans. These results demonstrate that native marsupials are not naïve to the peril humans pose, substantially expanding the taxonomic and geographic scope of the growing experimental evidence that wildlife worldwide generally perceive humans as the planet's most frightening predator. Introduced fallow deer fled humans, but not more than other predators, which we suggest may result from their being introduced. Our results point to both challenges concerning marsupial conservation and opportunities for exploiting fear of humans as a wildlife management tool.

Nose of dog, eye of elk, and wolf's liver: exploring the interconnectedness of Indigenous health and foraging among the Dukha reindeer herders of Mongolia.

Recent studies show that Traditional Ecological Knowledge (TEK) of Indigenous Peoples' food systems and the transfer of that knowledge to their children are critical to physical and mental health, promote the transfer of language and culture, and further resilience. The aim of this exploratory, medical ecological study was to document TEK about foraging and plant use among the Dukha, a small, semi-nomadic reindeer herding community in Northern Mongolia. Field research was conducted at multiple remote camps in 2019 and 2023. Methods included semi-structured, open-ended interviews (N = 16), photo identification, and participant observation (N = 34 families). Study Informants identified 27 foraged plants. Results placed Dukha foraging practices within a framework illustrating the interconnectedness of their worldview and cultural norms, including the physical and spiritual aspects of the landscape, with health, illness, and curing. Dukha curative knowledge of the flora aligns with both Traditional Mongolian Medicine (TMM) and scientific studies on healing and preventative care. Through foraging and plant use, the Dukha balance relationships with the land and spirits, benefit from better nutrition and physical activity, utilise healing qualities of indigenous flora, and foster the intergenerational transfer of language and traditional knowledge.

Kill wolves to save caribou? Sadly, it seems to work.

Some worry the findings will stall efforts to halt logging-the root cause of caribou population declines.

A panel of tetranucleotide STR markers as an alternative approach to forensic DNA identification of wolf and dog.

Commercial panels of microsatellite (STR) loci are focused on the use of DNA of the domestic dog (Canis lupus familiaris) and are often inapplicable for genotyping the DNA of the gray wolf (Canis lupus lupus). We propose a CPlex test system, including one hexa- and 12 tetranucleotide autosomal STR loci, as well as two sex loci, that is equally efficient in DNA identification of biological samples of the wolf and the dog. Analysis of molecular variance between samples revealed significant differentiation values (FST = 0.0784, p < 0.001), which allows to use the panel to differentiate wolf and dog samples. Population subdivision coefficients (θ-values) were calculated for each of the 13 STR loci of the developed test system. It was shown that the values of the genotype frequency for dogs and wolves, without and with considering the θ-value, differ by three orders of magnitude (for dogs 8.9 × 10-16 and 2.1 × 10-14 and for wolves 1.9 × 10-15 and 4.5 × 10-14, respectively). The use of population subdivision coefficients will allow to identify the most reliable results of an expert identification study and the power of exclusion provided by the STR loci of the CPlex test system makes it possible to achieve a reliable level of evidence in forensic DNA analysis of both wolves and dogs. The test system has been validated for use in forensic identification of the dog and wolf based on biological traces found at crime scenes, as well as for individual identification and establishing biological relationship of animals of these species.

Effects of lethal management on gray wolf pack persistence and reproduction in Wisconsin, USA.

Direct human-caused mortality accounts for about half of all large mammal mortality in North America. For social species like gray wolves (Canis lupus), the death of pack members can disrupt pack structure and cause pack dissolution, and mortality of breeding adults or wolves during reproduction and pup-rearing can decrease pup recruitment. We estimated minimum and maximum probability of wolf pack persistence in Wisconsin, USA, during biological years (15 April-14 April) 2011-2019 and evaluated the influence of pack size and legal harvest mortality on pack persistence during 2012-2014. Harvests comprised 75-161 mortalities within 194 monitored packs during 2012-2014, with 56-74% of packs having no wolves harvested each year. As an index of reproduction during 2013-2019, we also estimated the proportion of packs where pups responded to howl surveys. We evaluated the influence of pack size, legal harvest, and agency removal on reproduction during 2013-2015. Annual maximum pack persistence probability was uniformly high (0.95-1.00), and annual minimum pack persistence probability ranged from 0.86-0.98 with a possible decline during years of harvest. Reproduction was similar in years following harvest and agency removal (2013-2015, pup response = 0.27-0.40), and years without harvest or agency removal the year prior (2016-2019, pup response = 0.28-0.66). Pack size had a positive effect on pack persistence and reproduction. Total number of wolf mortalities and number of adult male and females removed did not influence pack persistence or reproduction. We suggest that low per-pack mortality, timing of harvest and agency removal, and harvest characteristics during 2012-2014 supported stable pack persistence and reproduction.

Relatedness-based mate choice and female philopatry: inbreeding trends of wolf packs in a human-dominated landscape.

Inbreeding can reduce offspring fitness and has substantial implications for the genetic diversity and long-term viability of populations. In social cooperative canids, inbreeding is conditioned by the geographic proximity between opposite-sex kin outside natal groups and the presence of related individuals in neighbouring groups. Consequently, challenges in moving into other regions where the species is present can also affect inbreeding rates. These can be particularly problematic in areas of high human density, where movement can be restricted, even for highly vagile species. In this study, we investigate the socio-ecological dynamics of Iberian wolf packs in the human-dominated landscape of Alto Minho, in northwest Portugal, where wolves exhibit a high prevalence of short-distance dispersal and limited gene flow with neighbouring regions. We hypothesise that mating occurs regardless of relatedness, resulting in recurrent inbreeding due to high kin encounter rates. Using data from a 10-year non-invasive genetic monitoring programme and a combination of relatedness estimates and genealogical reconstructions, we describe genetic diversity, mate choice, and dispersal strategies among Alto Minho packs. In contrast with expectations, our findings reveal relatedness-based mate choice, low kin encounter rates, and a reduced number of inbreeding events. We observed a high prevalence of philopatry, particularly among female breeders, with the most common breeding strategy involving the pairing of a philopatric female with an unrelated immigrant male. Overall, wolves were not inbred, and temporal changes in genetic diversity were not significant. Our findings are discussed, considering the demographic trend of wolves in Alto Minho and its human-dominated landscape.

A first exploratory comparison of the behaviour of wolves (Canis lupus) and wolf-dog hybrids in captivity.

Extensive introgression of genes from domesticated taxa may be a serious threat for the genomic integrity and adaptability of wild populations. Grey wolves (Canis lupus) are especially vulnerable to this phenomenon, but there are no studies yet assessing the potential behavioural effects of dog-introgression in wolves. In this study, we conducted a first systematic comparison of admixed (N = 11) and non-admixed (N = 14) wolves in captivity, focusing on their reaction to unfamiliar humans and novel objects, and the cohesiveness of their social groups. When exposed to unfamiliar humans in the experimental task, wolves were more vigilant, fearful and aggressive than admixed wolves, and less likely to approach humans, but also more likely to spend time in human proximity. When exposed to novel objects, wolves were more aggressive than admixed wolves, less likely to spend time in object proximity, and more likely to interact with objects, but also less vigilant and as fearful as admixed wolves. Finally, social networks were more cohesive in wolves than in admixed wolves. Although caution is needed when comparing groups of captive individuals with different life experiences, our study suggests that dog admixture may lead to important behavioural changes in wolves, with possible implications for conservation strategies.

Using meta-analysis to understand the impacts of dietary protein and fat content on the composition of fecal microbiota of domestic dogs (Canis lupus familiaris): A pilot study.

The interplay between diet and fecal microbiota composition is garnering increased interest across various host species, including domestic dogs. While the influence of dietary macronutrients and their associated microbial communities have been extensively reviewed, these reviews are descriptive and do not account for differences in microbial community analysis, nor do they standardize macronutrient content across studies. To address this, a meta-analysis was performed to assess the impact of dietary crude protein ("protein") and dietary crude fat ("fat") on the fecal microbiota composition in healthy dogs. Sixteen publications met the eligibility criteria for the meta-analysis, yielding a final data set of 314 dogs. Diets were classed as low, moderate, high, or supra in terms of protein or fat content. Sequence data from each publication were retrieved from public databases and reanalyzed using consistent bioinformatic pipelines. Analysis of community diversity indices and unsupervised clustering of the data with principal coordinate analysis revealed a small effect size and complete overlap between protein and fat levels at the overall community level. Supervised clustering through random forest analysis and partial least squares-discriminant analysis indicated alterations in the fecal microbiota composition at a more individual taxonomic level, corresponding to the levels of protein or fat. The Prevotellaceae Ga6A1 group and Enterococcus were associated with increasing levels of protein, while Allobaculum and Clostridium sensu stricto 13 were associated with increasing levels of fat. Interestingly, the random forest analyses revealed that Sharpea, despite its low relative abundance in the dog's fecal microbiome, was primarily responsible for the separation of the microbiome for both protein and fat. Future research should focus on validating and understanding the functional roles of these relatively low-abundant genera.

Diversity of Trichinella species in carnivores from Bosnia and Herzegovina.

BACKGROUND: In Bosnia and Herzegovina, domestic and wild carnivores represent a significant driver for the transmission and ecology of zoonotic pathogens, especially those of parasitic aetiology. Nevertheless, there is no systematic research of Trichinella species in animals that have been conducted in Bosnia and Herzegovina, even though trichinellosis is considered the most important parasitic zoonosis. The available results of the few studies carried out in Bosnia and Herzegovina are mainly related to the confirmation of parasitic larvae in the musculature of domestic pigs and wild boars or data related to trichinellosis in humans. The objective of our study was to present the findings of a comprehensive investigation into the species composition of Trichinella among 11 carnivorous species within the territory of Bosnia and Herzegovina, as follows: red fox (Vulpes vulpes), grey wolf (Canis lupus), brown bear (Ursus arctos), wildcat (Felis silvestris), pine marten (Martes martes), European badger (Meles meles), weasel (Mustela nivalis), European polecat (Mustela putorius), Eurasian lynx (Lynx lynx), but also dog (Canis lupus familiaris) and cat (Felis catus). RESULTS: In the period 2013-2023, carnivore musculature samples (n = 629), each consisting of 10 g of muscle tissue, were taken post-mortem and individually examined using the artificial digestion method. In the positive samples (n = 128), molecular genotyping and identification of parasitic larvae of Trichinella spp. were performed using a PCR-based technique up to the species/genotype level. Positive samples were used for basic PCR detection of the genus Trichinella (rrnS rt-PCR technique) and genotyping (rrnl-EVS rt-PCR technique). The Trichinella infection was documented for the first time in Bosnia and Herzegovina among red foxes, grey wolves, brown bears, dogs, badgers and Eurasian lynx, with a frequency rate of 20.3%. Additionally, the presence of T. britovi infection was newly confirmed in Bosnia and Herzegovina, marking the initial documented cases. Furthermore, both T. britovi and T. pseudospiralis infections were observed in the wildcat population, whereas T. britovi and T. spiralis infections were detected in pine martens. Consistent with previous research, our findings align particularly regarding carnivores, with data from other countries such as Germany, Finland, Romania, Poland and Spain, where T. britovi exhibits a wider distribution (62.5-100%) compared to T. spiralis (0.0-37.5%). T. britovi is more common among sylvatic carnivores (89.0%), while T. spiralis prevails in wild boars (62.0%), domestic swine (82.0%) and rodents (75.0%). CONCLUSION: The results of our study represent the first molecular identification of species of the genus Trichinella in Bosnia and Herzegovina. Additionally, our findings underscore the necessity for targeted epidemiological studies to thoroughly assess trichinellosis prevalence across diverse animal populations. Considering the relatively high frequency of trichinellosis infection in investigated animal species and its public health implications, there is an evident need for establishing an effective trichinellosis surveillance system in Bosnia and Herzegovina.

Does novelty influence the foraging decisions of a scavenger?

Acquiring knowledge about the environment is crucial for survival. Animals, often driven by their exploratory tendencies, gather valuable information regarding food resources, shelter, mating partners, etc. However, neophobia, or avoiding novel environmental stimuli, can constrain their exploratory behaviour. While neophobia can reduce potential predation risks, decreased exploratory behaviour resulting from it may limit the ability to discover highly rewarding resources. Dogs (Canis familiaris) living in semi-urban and urban environments as free-ranging populations, although subject to various selection forces, typically have negligible predation pressure. These dogs are scavengers in human-dominated environments; thus, selection against object-neophobia can provide benefits when searching for novel food resources. Although captive pack-living dogs are known to be less neophobic than their closest living ancestors, wolves (Canis lupus), little is known about free-ranging dogs' behavioural responses to novel objects, particularly in foraging contexts. Using an object choice experiment, we tested 259 free-ranging dogs from two age classes, adult and juvenile, to investigate their object-neophobia in a scavenging context. We employed a between-subject study design, providing dogs with a familiar and a potentially novel object, both baited with equal, hidden food items. Adult and juvenile dogs significantly inspected the novel object first compared to the familiar one, even when the hidden food item was partially visible. To validate these findings, we compared novel objects with different strengths of olfactory cues (baited vs. false-baited) and found that they were inspected comparably by adults and juveniles. No significant differences were found in the latencies to inspect the objects, suggesting that free-ranging dogs may still be cautious when exploring their environments. These results indicate that free-ranging dogs, evidently from an early ontogenetic phase, do not show object-neophobia, as demonstrated by their preference for novel over familiar food sources. We conclude that little to no constraint of neophobia on exploratory behaviour in semi-urban and urban-dwelling animals can guide foraging decision-making processes, providing adaptive benefits.

From wolves to humans: oral microbiome resistance to transfer across mammalian hosts.

The mammalian mouth is colonized by complex microbial communities, adapted to specific niches, and in homeostasis with the host. Individual microbes interact metabolically and rely primarily on nutrients provided by the host, with which they have potentially co-evolved along the mammalian lineages. The oral environment is similar across mammals, but the diversity, specificity, and evolution of community structure in related or interacting mammals are little understood. Here, we compared the oral microbiomes of dogs with those of wild wolves and humans. In dogs, we found an increased microbial diversity relative to wolves, possibly related to the transition to omnivorous nutrition following domestication. This includes a larger diversity of Patescibacteria than previously reported in any other oral microbiota. The oral microbes are most distinct at bacterial species or strain levels, with few if any shared between humans and canids, while the close evolutionary relationship between wolves and dogs is reflected by numerous shared taxa. More taxa are shared at higher taxonomic levels including with humans, supporting their more ancestral common mammalian colonization followed by diversification. Phylogenies of selected oral bacterial lineages do not support stable human-dog microbial transfers but suggest diversification along mammalian lineages (apes and canids). Therefore, despite millennia of cohabitation and close interaction, the host and its native community controls and limits the assimilation of new microbes, even if closely related. Higher resolution metagenomic and microbial physiological studies, covering a larger mammalian diversity, should help understand how oral communities assemble, adapt, and interact with their hosts.IMPORTANCENumerous types of microbes colonize the mouth after birth and play important roles in maintaining oral health. When the microbiota-host homeostasis is perturbed, proliferation of some bacteria leads to diseases such as caries and periodontitis. Unlike the gut microbiome, the diversity of oral microbes across the mammalian evolutionary space is not understood. Our study compared the oral microbiomes of wild wolves, dogs, and apes (humans, chimpanzees, and bonobos), with the aim of identifying if microbes have been potentially exchanged between humans and dogs as a result of domestication and cohabitation. We found little if any evidence for such exchanges. The significance of our research is in finding that the oral microbiota and/or the host limit the acquisition of exogenous microbes, which is important in the context of natural exclusion of potential novel pathogens. We provide a framework for expanded higher-resolution studies across domestic and wild animals to understand resistance/resilience.

Epidemiological survey of infectious agents in free-ranging maned wolves (Chrysocyon brachyurus) in Northeastern Brazil.

Infectious diseases are one of the most concerning threats to maned wolves (Chrysocyon brachyurus) due to the potential impact on free-ranging populations. The species is currently classified as vulnerable according to the national list of threatened species and occurs mainly in open habitats, such as the Cerrado, a tropical savannah, which comprises its main distribution area in Brazil. In the northeastern region, it occurs in the Cerrado of Bahia, Piauí, Maranhão, and Tocantins states. Therefore, this study aimed to investigate the occurrence of infectious agents in Chrysocyon brachyurus through an epidemiological assessment of free-ranging individuals in western Bahia, specifically in the Barreiras microregion, a Cerrado area intensely fragmented and anthropized by agricultural activity. Eleven specimens were evaluated for serological titration, antigen research, and genetic material research for canine distemper virus (CDV), canine parvovirus (CPV), adenovirus-canine-type 1 (CAdV-1), canine coronavirus (CCoV), Leptospira interrogans and Toxoplasma gondii from 2020 to 2022. In addition to maned wolves, domestic dogs were also evaluated and tested. All maned wolves (100%) evaluated by the dot-ELISA technique exhibited immunoglobulin M (IgM) and seven (64%) exhibited immunoglobulin G (IgG) against CDV and CPV, while 100% exhibited IgG against CDV when using the immunochromatographic technique. Regarding CAdV-1, 90% were seropositive for IgG, while 64% exhibited IgG against T. gondii. Nine dogs from the region were also sampled, and all (100%) exhibited IgM and IgG against CDV and CPV. For IgG against T. gondii and against CAdV-1, 90% of the animals were seropositive. Molecular evaluation yielded negative results for all maned wolves and dogs assessed for CAdV-1, CDV, and T. gondii, as well as the CCoV antigen. These data indicate the occurrence of viral agents and Toxoplasma gondii in maned wolves and dogs, suggesting circulation in both populations.

Applying XGBoost and SHAP to Open Source Data to Identify Key Drivers and Predict Likelihood of Wolf Pair Presence.

Wolves have returned to Germany since 2000. Numbers have grown to 209 territorial pairs in 2021. XGBoost machine learning, combined with SHAP analysis is applied to predict German wolf pair presence in 2022 for 10 × 10 km grid cells. Model input consisted of 38 variables from open sources, covering the period 2000 to 2021. The XGBoost model predicted well, with 0.91 as the AUC. SHAP analysis ranked the variables: distance to the closest neighboring wolf pair was the main driver for a grid cell to become occupied by a wolf pair. The clustering tendency of related wolves seems to be an important explanatory factor here. Second was the percentage of wooded area. The next eight variables related to wolf presence in the preceding year, except at fifth, eighth and tenth position in the total order: human density (square root) in the grid, percentage arable land and road density respectively. Other variables including the occurrence of wild prey were the weakest predictors. The SHAP analysis also provided crucial added value in identifying a variable that had threshold values where its contribution to the prediction changed from positive to negative or vice versa. For instance, low density of people increased the probability of wolf pair presence, whereas a high density decreased this probability. Cumulative lift techniques showed that the model performed almost four times better than random prediction. The combination of XGBoost, SHAP and cumulative lift techniques is new in wolf management and conservation, allowing for the focusing of educational and financial resources.

Hybrid Improved Grey Wolf Support Vector Regression Algorithm for Modeling Solubilities of APIs in Pure Ionic Liquids: σ-Profile Descriptors.

The objective of this study was to model the solubility of active pharmaceutical ingredients (APIs) in different ionic liquids (ILs) based on the σ-moments of cations, anions, and APIs that were used as molecular descriptors calculated using the σ-profiles of three categories of descriptors based on conductor-like screening model for real solvents. The database of 83 API-ILs systems composed of 14 APIs, 12 cations, and 7 anions (25 ILs combinations) was collected as 850 data points at different temperature ranges. A hybrid Improved Grey Wolf Support vector regression, abbreviated as I-GWO-SVR(r), algorithm was selected as the learning method. Based on a comprehensive comparison with 11 different models, various statistical factors, and graphical analyses, including an external validation test, analysis of variance (ANOVA), and sensitivity analysis, the capability and validity of the proposed approach have been assessed and verified. The overall study confirmed that the proposed new model provided the best results in terms of predicting the solubility of APIs in ILs.

Sero-epidemiological investigation on Toxoplasma gondii infection in Apennine wolf (Canis lupus italicus) and wild boar (Sus scrofa) in Italy.

BACKGROUND: The wild boar (Sus scrofa) and the Apennine wolf (Canis lupus italicus) are two wild species that have both increased their presence in the Italian territory, albeit in varying numbers. They can be occasionally found in peri-urban areas as well. Both of these species can serve as intermediate hosts for Toxoplasma gondii, as they can become infected either through the consumption of oocysts found in water, soil, or on vegetables, or through the ingestion of meat containing bradyzoites. Consequently, these animals can be regarded as key indicators of Toxoplasma presence in the wild or peri-urban environment. In our study, we examined a total of 174 wild boar meat juice and 128 wolf sera from Italy for the detection of T. gondii IgG using the indirect fluorescent antibody test (IFAT). RESULTS: The results showed that 40 (22.6%) of the wild boar meat juice and 34 (26.6%) of the wolf serum samples tested positive. Interestingly, there were no significant differences in seropositivity with respect to gender, age group, or the region of origin in both species. CONCLUSIONS: Overall the results indicate a moderate exposure in both the species under investigation, highlighting the spread of T. gondii in sylvatic and periurban environments. The prevalence of T. gondii in wild boar is consistent with findings from other studies conducted in Europe. Our study, with a considerably larger sample size compared to the available research in European context, provides valuable data on the seroprevalence of T. gondii in wolves.

Bioactive triterpenoid compounds of Poria cocos (Schw.) Wolf in the treatment of diabetic ulcers via regulating the PI3K-AKT signaling pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Diabetic ulcers represent a chronic condition characterized by prolonged hyperglycemia and delayed wound healing, accompanied by endocrine disorders, inflammatory responses, and microvascular damage in the epidermal tissue, demanding effective clinical treatment approaches. For thousands of years, ancient Chinese ethnopharmacological studies have documented the use of Poria cocos (Schw.) Wolf in treating diabetic ulcers. Recent research has substantiated the diverse pharmacological effects of Poria cocos (Schw.) Wolf, including its potential to alleviate hyperglycemia and exhibit anti-inflammatory, antioxidant, and immune regulatory properties, which could effectively mitigate diabetic ulcer symptoms. Furthermore, being a natural medicine, Poria cocos (Schw.) Wolf has demonstrated promising therapeutic effects and safety in the management of diabetic ulcers, holding significant clinical value. Despite its potential clinical efficacy and applications in diabetic ulcer treatment, the primary active components and underlying pharmacological mechanisms of Poria cocos (Schw.) Wolf remains unclear. Further investigations are imperative to establish a solid foundation for drug development in this domain. AIM OF THE STUDY AND MATERIALS AND METHODS: In this study, we aimed to identify the active compounds and potential targets of Poria cocos (Schw.) Wolf using UHPLC-Q-TOF-MS and TCMSP databases. Additionally, we attempt to identify targets related to diabetic ulcers. Following enrichment analysis, a network of protein-protein interactions was constructed to identify hub genes based on the common elements between the two datasets. To gain insights into the binding activities of the hub genes and active ingredients, molecular docking analysis was employed. Furthermore, to further validate the therapeutic effect of Poria cocos (Schw.) Wolf, we exerted in vitro experiments using human umbilical vein vascular endothelial cells and human myeloid leukemia monocytes (THP-1). The active ingredient of Poria cocos (Schw.) Wolf was applied in these experiments. Our investigations included various assays, such as CCK-8, scratch test, immunofluorescence, western blotting, RT-PCR, and flow cytometry, to explore the potential of Poria cocos (Schw.) Wolf triterpenoid extract (PTE) in treating diabetic ulcers. RESULTS: The findings here highlighted PTE as the primary active ingredient in Poria cocos (Schw.) Wolf. Utilizing network pharmacology, we identified 74 potential targets associated with diabetic ulcer treatment for Poria cocos (Schw.) Wolf, with five hub genes (JUN, MAPK1, STAT3, AKT1, and CTNNB1). Enrichment analysis revealed the involvement of multiple pathways in the therapeutic process, with the PI3K-AKT signaling pathway showing significant enrichment. Through molecular docking, we discovered that relevant targets within this pathway exhibited strong binding with the active components of Poria cocos (Schw.) Wolf. In vitro experiments unveiled that PTE (10 mg/L) facilitated the migration of human umbilical vein vascular endothelial cells (P < 0.05). PTE also increased the expression of CD31 and VEGF mRNA (P < 0.05) while activating the expressions of p-PI3K and p-AKT (P < 0.05). Moreover, PTE demonstrated its potential by reducing the expression of IL-1ß, IL-6, TNF-α, and NF-κB mRNA in THP-1 (P < 0.05) and fostering M2 macrophage polarization. These results signify the potential therapeutic effects of PTE in treating diabetic ulcers, with its beneficial actions mediated through the PI3K-AKT signaling pathway. CONCLUSIONS: PTE is the main active ingredient in Poria cocos (Schw.) Wolf that exerts therapeutic effects. Through PI3K-AKT signaling pathway activation and inflammatory response reduction, PTE promotes angiogenesis, thereby healing diabetic ulcers.

Cougars, wolves, and humans drive a dynamic landscape of fear for elk.

To manage predation risk, prey navigate a dynamic landscape of fear, or spatiotemporal variation in risk perception, reflecting predator distributions, traits, and activity cycles. Prey may seek to reduce risk across this landscape using habitat at times and in places when predators are less active. In multipredator landscapes, avoiding one predator could increase vulnerability to another, making the landscape of fear difficult to predict and navigate. Additionally, humans may shape interactions between predators and prey, and induce new sources of risk. Humans can function as a shield, providing a refuge for prey from human-averse carnivores, and as a predator, causing mortality through hunting and vehicle collisions and eliciting a fear response that can exceed that of carnivores. We used telemetry data collected between 2017 and 2021 from 63 Global Positioning System-collared elk (Cervus canadensis), 42 cougars (Puma concolor), and 16 wolves (Canis lupus) to examine how elk habitat selection changed in relation to carnivores and humans in northeastern Washington, USA. Using step selection functions, we evaluated elk habitat use in relation to cougars, wolves, and humans, diel period (daytime vs. nighttime), season (summer calving season vs. fall hunting season), and habitat structure (open vs. closed habitat). The diel cycle was critical to understanding elk movement, allowing elk to reduce encounters with predators where and when they would be the largest threat. Elk strongly avoided cougars at night but had a near-neutral response to cougars during the day, whereas elk avoided wolves at all times of day. Elk generally used more open habitats where cougars and wolves were most active, rather than altering the use of habitat structure depending on the predator species. Elk avoided humans during the day and ~80% of adult female mortality was human caused, suggesting that humans functioned as a "super predator" in this system. Simultaneously, elk leveraged the human shield against wolves but not cougars at night, and no elk were confirmed to have been killed by wolves. Our results add to the mounting evidence that humans profoundly affect predator-prey interactions, highlighting the importance of studying these dynamics in anthropogenic areas.