Anal Gland Secretion Research Articles

Characterization of the microbiome and volatile compounds in anal gland secretions from domestic cats (Felis catus) using metagenomics and metabolomics

Many mammals rely on volatile organic chemical compounds (VOCs) produced by bacteria for their communication and behavior, though little is known about the exact molecular mechanisms or bacterial species that are responsible. We used metagenomic sequencing, mass-spectrometry based metabolomics, and culturing to profile the microbial and volatile chemical constituents of anal gland secretions in twenty-three domestic cats (Felis catus), in attempts to identify organisms potentially involved in host odor production. We found that the anal gland microbiome was dominated by bacteria in the genera Corynebacterium, Bacteroides, Proteus, Lactobacillus, and Streptococcus, and showed striking variation among individual cats. Microbiome profiles also varied with host age and obesity. Metabolites such as fatty-acids, ketones, aldehydes and alcohols were detected in glandular secretions. Overall, microbiome and metabolome profiles were modestly correlated (r = 0.17), indicating that a relationship exists between the bacteria in the gland and the metabolites produced in the gland. Functional analyses revealed the presence of genes predicted to code for enzymes involved in VOC metabolism such as dehydrogenases, reductases, and decarboxylases. From metagenomic data, we generated 85 high-quality metagenome assembled genomes (MAGs). Of importance were four MAGs classified as Corynebacterium frankenforstense, Proteus mirabilis, Lactobacillus johnsonii, and Bacteroides fragilis. They represent strong candidates for further investigation of the mechanisms of volatile synthesis and scent production in the mammalian anal gland.

A New Webbing Aberoptus Species from South Africa Provides Insight in Silk Production in Gall Mites (Eriophyoidea)

Arthropods include a high diversity of lineages adapted for silk production. Several species of microscopic phytophagous mites of the hyperdiverse superfamily Eriophyoidea spin web; however, the origin of their silk is unknown. We described a new web-spinning mite, Aberoptus schotiae n. sp., collected from leaves of Schotia brachypetala (Fabaceae) in South Africa and showed that it has a complex life cycle, including two morphotypes of adults and nymphs. Molecular phylogenetic analyses and 28S sequence comparison showed conspecificity of heteromorphic females and rejected synonymy of Aberoptus and Aceria proposed by previous authors. For the first time, we provided SEM images of the web nests and, using a set of different microscopic techniques, described the silk-producing anal secretory apparatus (ASA) of Aberoptus. It comprises two pairs of anal glands (hypertrophied in web-spinning females), three cuticular sacs and a rectal tube leading to the anal opening. This is a unique case (analogy) of anal silk secretion in Chelicerata previously reported only in Serianus (Pseudoscorpiones). Recent findings of rudimentary ASA in distant eriophyoid lineages and the results of this study transform the current paradigm of exoticism of web-spinning eriophyoid taxa into the concept of synapomorphic specialization of the hindgut for excreting the anal gland secretions in Eriophyoidea.

Identification of the parasite, Trypanosoma cruzi, in multiple tissues of epidemiological significance in the Virginia opossum (Didelphis virginiana): Implications for environmental and vertical transmission routes.

Trypanosoma cruzi, a parasitic protozoan, is endemic to the Americas and the causative agent of Chagas disease in humans. In South America, opossums facilitate transmission via infected anal gland secretions in addition to transmission via triatomine vectors. In North America, the Virginia opossum is a reservoir host for the parasite with transmission routes that are not clearly defined. The unique biology of this marsupial provides the opportunity to investigate vertical transmission in this wildlife species in situ. Our objectives were to investigate alternative routes of transmission that may facilitate spillover into other species and to determine if vertical transmission was evident. Virginia opossums were sampled at 10 trapping locations over a 10-month period in a 5-county region of north central Florida. Peripheral blood, fecal swabs, and anal gland secretions were collected from each adult individual, and peripheral blood was collected from joey opossums. Total DNA was extracted from each collected sample type, and T. cruzi infected individuals and the infecting Discrete Typing Unit (DTU) were identified using real time PCR methods. Adult Virginia opossums (n = 112) were infected with T. cruzi (51.8%, 95% CI [42.6-60.8%]) throughout the sampled period and at each location. T. cruzi DNA was found in each of the three biological sample types. Vertical transmission of T. cruzi was inferred in one litter of mother-dependent (n = 20, 5.0%, 95% CI [0.9-23.6%]) joey opossums where 2 joeys from this same litter were rtPCR positive for T. cruzi. We inferred vertical transmission from mother to neonate which may serve to amplify the prevalence of T. cruzi in adult Virginia opossums. T. cruzi DNA was detected in the anal gland secretions of Virginia opossums. Infected anal gland secretions suggest a possible environmental route of transmission for T. cruzi via the deposition of contaminated feces and spraint at wildlife latrines. Only DTU1 was identified in the sampled population which is consistent with human autochthonous cases in the United States.

The vomeronasal system in semiaquatic beavers.

In contrast to the main olfactory system that detects volatile chemicals in the nasal air, the vomeronasal system can detect nonvolatile chemicals as well as volatiles. In the vomeronasal system, chemicals are perceived by the vomeronasal organ (VNO) projecting axons to the accessory olfactory bulb (AOB). Beavers (Castor spp.) are semiaquatic mammals that have developed chemical communication. It is possible that the beaver's anal gland secretions, nonvolatile and insoluble substances, may work as a messenger in the water and that beavers may detect the nonvolatile chemicals floating on the water surface via the VNO. The present study aimed to clarify the specificities of the beaver vomeronasal system by histologically and immunohistochemically analyzing the VNO and AOB of 12 Eurasian beavers (C. fiber). The VNO directly opened to the nasal cavity and was independent of a narrow nasopalatine duct connecting the oral and nasal cavities. The VNO comprised soft tissues including sensory and nonsensory epithelium, glands, a venous sinus, an artery, as well as cartilage inner, and bone outer enclosures. The AOB had distinct six layers, and anti-G protein α-i2 and α-o subunits were, respectively, immunoreactive in rostral and caudal glomeruli layers indicating expressions of V1Rs and V2Rs. According to gene repertories analysis, the beavers had 23 and six intact V1R and V2R genes respectively. These findings suggested that beavers recognize volatile odorants and nonvolatile substances using the vomeronasal system. The beaver VNO was developed as well as in other rodents, and it had two specific morphological features, namely, disadvantaged contact with the oral cavity because of a tiny nasopalatine duct, and a double bone and cartilage envelope. Our results highlight the importance of the vomeronasal system in beaver chemical communication and support the possibility that beavers can detect chemicals floating on the water surface via the VNO.

Influencing Elimination Location in the Domestic Cat: A Semiochemical Approach.

Simple SummaryIn the domestic cat, elimination at an inappropriate location is considered by cat owners and non-cat owners as an undesirable behaviour. The aim of the study was to assess the effects of a composition derived from cat anal glands on the elimination behaviour of domestic cats in a cattery setting. The study was conducted in four catteries housing 33 cats, using 37 litter trays. The data collection lasted two weeks, each litter tray receiving one treatment on the first week and the other treatment on the second week. The parameters studied included daily elimination (urine plus stools) weight, urine weight, stool weight, elimination type and urine/stool quantity scoring. Four out of the six parameters studied showed a treatment effect, consistently in favour of cats defecating significantly less in the litter trays sprayed with the treatment versus litter trays sprayed with the control. These results demonstrate that a composition originating from cat anal glands influences cats’ defecation location.In the domestic cat, elimination at an inappropriate location is considered by cat owners and non-cat owners as an undesirable behaviour. The aim of the study was to assess the effect of a semiochemical formulation, reconstituted volatile fraction of cat anal gland secretions on the elimination behaviour of domestic cats. The study was conducted in four catteries, which housed 33 cats, using 37 litter trays and followed a randomised crossover design using the litter tray as the experimental unit. The parameters studied included daily elimination (urine plus stools) weight, urine weight, stool weight, elimination type and urine/stool quantity scoring. The parameters were analysed using GLMM with SAS 9.4 software. Four out of the six parameters studied showed a treatment effect, consistently in favour of cats defecating significantly less in the litter trays sprayed with the treatment versus litter trays sprayed with the control (elimination weight p = 0.0199; elimination type p = 0.0251; stool weight p = 0.0005 and stool quantity p = 0.003). These results demonstrate that an intraspecific semiochemical message originating from cat anal glands influences cats’ defecation location.

Odorant-binding proteins in canine anal sac glands indicate an evolutionarily conserved role in mammalian chemical communication

BackgroundChemical communication is an important aspect of the behavioural ecology of a wide range of mammals. In dogs and other carnivores, anal sac glands are thought to convey information to conspecifics by secreting a pallet of small volatile molecules produced by symbiotic bacteria. Because these glands are unique to carnivores, it is unclear how their secretions relate to those of other placental mammals that make use of different tissues and secretions for chemical communication. Here we analyse the anal sac glands of domestic dogs to verify the secretion of proteins and infer their evolutionary relationship to those involved in the chemical communication of non-carnivoran mammals.ResultsProteomic analysis of anal sac gland secretions of 17 dogs revealed the consistently abundant presence of three related proteins. Homology searches against online databases indicate that these proteins are evolutionary related to ‘odorant binding proteins’ (OBPs) found in a wide range of mammalian secretions and known to contribute to chemical communication. Screening of the dog’s genome sequence show that the newly discovered OBPs are encoded by a single cluster of three genes in the pseudoautosomal region of the X-chromosome. Comparative genomic screening indicates that the same locus is shared by a wide range of placental mammals and that it originated at least before the radiation of extant placental orders. Phylogenetic analyses suggest a dynamic evolution of gene duplication and loss, resulting in large gene clusters in some placental taxa and recurrent loss of this locus in others. The homology of OBPs in canid anal sac glands and those found in other mammalian secretions implies that these proteins maintained a function in chemical communication throughout mammalian evolutionary history by multiple shifts in expression between secretory tissues involved in signal release and nasal mucosa involved in signal reception.ConclusionsOur study elucidates a poorly understood part of the biology of a species that lives in close association with humans. In addition, it shows that the protein repertoire underlying chemical communication in mammals is more evolutionarily stable than the variation of involved glands and tissues would suggest.

Dogs can scent-match individual Eurasian beavers from their anal gland secretion

Dogs Canis lupus familiaris are increasingly being used in wildlife conservation studies, due to their extensive ofactory capabilities. Dogs are a useful tool for species detection, species discrimination (or subspecies), and scent-matching of individuals within a species. Scent-matching can reduce or eliminate the need for expensive genotyping of obtained biological samples. We investigated the potential use of dogs to scent-match individual Eurasian beavers Castor fiber via anal gland secretion (AGS) samples, in 30 double blind floor platform experiments. We hypothesised that dogs can scent-match individual beavers when presented with AGS from different beavers of both sexes. We showed that dogs were able to scent-match individual beavers with average accuracy of 88.9%, sensitivity of 66.7% and specificity of 93.3%. Our results suggest that scent-matching dogs may be used as a reliable additional method to DNA analysing of biological samples to improve accuracy of individual beaver detection, and a better alternative than live-trapping/capturing in monitoring of specific beavers in e.g. a reintroduction project.

Fears from the past? The innate ability of dogs to detect predator scents

Throughout the animal kingdom, antipredator mechanisms are an evolutionary driving force to enable the survival of species classified as prey. Information regarding a predator’s location can be determined through chemosensory cues from urine, faeces, visual and/or acoustic signals and anal gland secretions; and in several lab and field-based studies it has been seen that these cues mediate behavioural changes within prey species. These behaviours are often linked to fear and avoidance, which will in turn increase the prey’s survival rate. In many studies dogs (Canis lupus familiaris) have been used as a predator species, however, no research has addressed a dog’s innate ability to detect predator scents, hence the rationale behind this study. We assessed the innate ability of the untrained domestic dog to detect faecal scents of wild Eurasian brown bear (Ursus arctos arctos) and European lynx (Lynx lynx). The study monitored 82 domestic dogs across the UK and Norway. The dogs were exposed to the two predator faecal scents from Eurasian brown bear and European lynx, a herbivore faecal scent of Eurasian beaver (Castor fiber) and water control. Measurements were taken upon the time spent within a 40 cm radius of each scent and changes in the dog’s heart rate when within this 40 cm radius. We found dogs spent a decreased length of time around the predator scents and had an increased heart rate in relation to their basal heart rate. We conclude that dogs can innately sense predator scents of brown bear and lynx and elicit fear towards these odours, as shown through behavioural and physiological changes.

Trypanosoma cruzi infections and associated pathology in urban-dwelling Virginia opossums (Didelphis virginiana).

Trypanosoma cruzi, a zoonotic protozoan parasite, infects a wide range of mammals. The southern United States has endemic sylvatic transmission cycles maintained by several species of wildlife and domestic dogs. We hypothesized that urban-dwelling opossums (Didelphis virginiana) in South Texas are infected with T. cruzi, and that tissue pathology would be associated with infection. In 2017, we collected blood, heart tissue and anal gland secretions from 100 wild opossums across three seasons that were trapped by animal control in South Texas. In addition, anal gland tissue and intercostal muscle were collected from 43 of the 100 opossums for which time allowed the extra tissue collection. All blood, tissue, and secretion samples were screened for T. cruzi DNA using qPCR with confirmation of positive status achieved through one or more additional PCR assays, including a qPCR to determine the parasite discrete typing unit (DTU). T. cruzi DNA was detected in at least one tissue of 15% of the opossums sampled: blood clot (9%), heart tissue (10%), anal gland secretions (12%), intercostal muscle (16.3%), and anal gland tissue (11.6%). Infection was detected in two or more different tissue types in nine of the opossums. The 35 tissues for which parasite DTU was determined were exclusively ‘Tcl’- a DTU previously associated with locally-acquired human disease in the United States. T. cruzi-positive opossums were nearly 14 times more likely to exhibit significant heart lesions on histopathology (lympoplasmacytic inflammation±fibrosis) when compared to negative opossums (OR = 13.56, CI = 1.23–751.28, p-value = 0.03). Three triatomines were opportunistically collected from the study site, of which two were infected (66.7%), and bloodmeal analysis revealed canine, opossum, and human bloodmeals. Given the presence of parasite in opossum blood, unique potential for shedding of parasite in anal glad secretions, and evidence of vectors feeding on opossums, it is likely that opossums serve as wild reservoirs around urban dwellings in South Texas.

Knowing Me, Knowing You: Anal Gland Secretion of European Badgers (Meles meles) Codes for Individuality, Sex and Social Group Membership

European badgers, Meles meles, are group-living in the UK, and demarcate their ranges with shared latrines. As carnivores, badgers possess paired anal glands, but olfactory information on the content of badger anal gland secretion (AGS) is largely uninvestigated. Here, we examined the volatile organic compounds (VOCs) of AGS samples from 57 free-living badgers using solid-phase microextraction (SPME) and gas chromatography—mass spectrometry. AGS was rich in alkanes (C7–C15, 14.3% of identified compounds), aldehydes (C5–C14, 9.7%), phenols (C6–C15, 9.5%), alcohols (C5–C10, 7.3%), aromatic hydrocarbons (C6–C13, 6.8%), ketones (C6–C13, 6.3%) and carboxylic acids (C3–C12, 5.6%) and contained a variety of esters, sulfurous and nitrogenous compounds, and ethers. The number of VOCs per profile ranged from 20 to 111 (mean = 65.4; ± 22.7 SD), but no compound was unique for any of the biological categories. After normalization of the raw data using Probabilistic Quotient Normalization, we produced a resemblance matrix by calculating the Euclidian distances between all sample pairs. PERMANOVA revealed that AGS composition differs between social groups, and concentration and complexity in terms of number of measurable VOCs varies between seasons and years. AGS VOC profiles encode individual identity, sex and vary with female reproductive state, indicating an important function in intraspecific communication. Because AGS is excreted together with fecal deposits, we conclude that chemical complexity of AGS enables particularly latrine-using species, such as badgers, to advertise more complex individual-specific information than in feces alone.

An Electrophilic Natural Product Provides a Safe and Robust Odor Neutralization Approach To Counteract Malodorous Organosulfur Metabolites Encountered in Skunk Spray.

The anal secretions of skunks comprise several types of malodorous organosulfur compounds. The pungent metabolites are used defensively by skunks to repel threats posed by predators, and in many parts of the world, those perceived threats include humans and their pets. The extremely low thresholds for detection of the organosulfur metabolites make efforts to "de-skunk" people, animals, and clothing a process fraught with many challenges. The fungal-derived metabolite pericosine A (4) is a promiscuous yet stabile electrophilic compound that we propose is used by some fungi as a novel form of chemical defense. Our investigations have indicated that pericosine A readily reacts with skunk-spray secretions to transform them into odorless products. Mechanistic and computational studies suggested that pericosine A and its synthetic analogues react via SN2'-type mechanisms with thiols and thioacetates under aqueous conditions to generate stable thioethers. Testing revealed that pericosine A did not cause skin or eye irritation and was highly effective at deodorizing skunk anal gland secretions when formulated to include adjunctive cosmetic ingredients.

Anal scent gland secretions inform on sexual maturity, sex and social status in the Alpine marmot, Marmota marmota (Rodentia: Sciuridae): a role in intrasexual competition in cooperative breeders?

Anal scent gland secretions inform on sexual maturity, sex and social status in the Alpine marmot, Marmota marmota (Rodentia: Sciuridae): a role in intrasexual competition in cooperative breeders?

Pathology of the Emerging Mycobacterium tuberculosis Complex Pathogen, Mycobacterium mungi, in the Banded Mongoose ( Mungos mungo).

Wild banded mongooses ( Mungos mungo) in northeastern Botswana and northwest Zimbabwe are infected with a novel Mycobacterium tuberculosis complex (MTC) pathogen, Mycobacterium mungi. We evaluated gross and histologic lesions in 62 infected mongooses (1999-2017). Many tissues contained multifocal irregular, lymphohistiocytic to granulomatous infiltrates and/or multifocal or coalescing noncaseating to caseating granulomas with variable numbers of intralesional acid-fast bacilli. Over one-third of nasal turbinates examined had submucosal lymphohistiocytic to granulomatous infiltrates, erosion and ulceration of the nasal mucosa, bony remodeling, and nasal distortion. Similar inflammatory cell infiltrates expanded the dermis of the nasal planum with frequent ulceration. However, even in cases with intact epidermis, acid-fast bacilli were present in variable numbers among dermal infiltrates and on the epidermal surface among desquamated cells and debris, most commonly in small crevices or folds. In general, tissue involvement varied among cases but was highest in lymph nodes (50/54, 93%), liver (39/53, 74%), spleen (37/51, 73%), and anal glands/sacs (6/8, 75%). Pulmonary lesions were present in 67% of sampled mongooses (35/52) but only in advanced disseminated disease. The pathological presentation of M. mungi in the banded mongoose is consistent with pathogen shedding occurring through scent-marking behaviors (urine and anal gland secretions) with new infections arising from contact with these contaminated olfactory secretions and percutaneous movement of the pathogen through breaks in the skin, nasal planum, and/or skin of the snout. Given the character and distribution of lesions and the presence of intracellular acid-fast bacilli, we hypothesize that pathogen spread occurs within the body through a hematogenous and/or lymphatic route. Features of prototypical granulomas such as multinucleated giant cells and peripheral fibrosis were rarely present in affected mongooses. Acid-fast bacilli were consistently found intracellularly, even in regions of necrosis. The mongoose genome has a unique deletion (RD1mon) that includes part of the encoding region for PPE68 (Rv3873), a gene co-operonic with PE35. These proteins can influence the host's cellular immune response to mycobacterial infections, and it remains uncertain how this deletion might contribute to observed patterns of pathology. M. mungi infection in banded mongooses is characterized by both a unique transmission and exposure route, as well as accompanying pathological features, providing an opportunity to increase our understanding of MTC pathogenesis across host-pathogen systems.

Social odours covary with bacterial community in the anal secretions of wild meerkats

The fermentation hypothesis for animal signalling posits that bacteria dwelling in an animal’s scent glands metabolize the glands’ primary products into odorous compounds used by the host to communicate with conspecifics. There is, however, little evidence of the predicted covariation between an animal’s olfactory cues and its glandular bacterial communities. Using gas chromatography-mass spectrometry, we first identified the volatile compounds present in ‘pure’ versus ‘mixed’ anal-gland secretions (‘paste’) of adult meerkats (Suricata suricatta) living in the wild. Low-molecular-weight chemicals that likely derive from bacterial metabolism were more prominent in mixed than pure secretions. Focusing thereafter on mixed secretions, we showed that chemical composition varied by sex and was more similar between members of the same group than between members of different groups. Subsequently, using next-generation sequencing, we identified the bacterial assemblages present in meerkat paste and documented relationships between these assemblages and the host’s sex, social status and group membership. Lastly, we found significant covariation between the volatile compounds and bacterial assemblages in meerkat paste, particularly in males. Together, these results are consistent with a role for bacteria in the production of sex- and group-specific scents, and with the evolution of mutualism between meerkats and their glandular microbiota.

Emerging Tuberculosis Pathogen Hijacks Social Communication Behavior in the Group-Living Banded Mongoose (Mungos mungo).

ABSTRACTAn emerging Mycobacterium tuberculosis complex (MTC) pathogen, M. mungi, infects wild banded mongooses (Mungos mungo) in Northern Botswana, causing significant mortality. This MTC pathogen did not appear to be transmitted through a primary aerosol or oral route. We utilized histopathology, spoligotyping, mycobacterial interspersed repetitive units-variable number of tandem repeats (MIRU-VNTR), quantitative PCR (qPCR), and molecular markers (regions of difference [RDs] from various MTC members, including region of difference 1 [RD1] from M. bovis BCG [RD1BCG], M. microti [RD1mic], and M. pinnipedii [RD1seal], genes Rv1510 [RD4], Rv1970 [RD7], Rv3877/8 [RD1], and Rv3120 [RD12], insertion element IS1561, the 16S RNA gene, and gene Rv0577 [cfp32]), including the newly characterized mongoose-specific deletion in RD1 (RD1mon), in order to demonstrate the presence of M. mungi DNA in infected mongooses and investigate pathogen invasion and exposure mechanisms. M. mungi DNA was identified in 29% of nasal planum samples (n = 52), 56% of nasal rinses and swabs (n = 9), 53% of oral swabs (n = 19), 22% of urine samples (n = 23), 33% of anal gland tissue (n = 18), and 39% of anal gland secretions (n = 44). The occurrence of extremely low cycle threshold values obtained with qPCR in anal gland and nasal planum samples indicates that high levels of M. mungi can be found in these tissue types. Histological data were consistent with these results, suggesting that pathogen invasion occurs through breaks in the nasal planum and/or skin of the mongoose host, which are in frequent contact with anal gland secretions and urine during olfactory communication behavior. Lesions in the lung, when present, occurred only with disseminated disease. No environmental sources of M. mungi DNA could be found. We report primary environmental transmission of an MTC pathogen that occurs in association with social communication behavior.

Behavioral Responses of CD-1 Mice to Six Predator Odor Components.

Mammalian prey species are able to detect predator odors and to display appropriate defensive behavior. However, there is only limited knowledge about whether single compounds of predator odors are sufficient to elicit such behavior. Therefore, we assessed if predator-naïve CD-1 mice (n = 60) avoid sulfur-containing compounds that are characteristic components of natural predator odors and/or display other indicators of anxiety. A 2-compartment test arena was used to assess approach/avoidance behavior, general motor activity, and the number of fecal pellets excreted when the animals were presented with 1 of 6 predator odor components in one compartment and a blank control in the other compartment. We found that 2 of the 6 predator odor components (2-propylthietane and 3-methyl-1-butanethiol) were significantly avoided by the mice. The remaining 4 predator odor components (2,2-dimethylthietane, 3-mercapto-3-methylbutan-1-ol, 3-mercapto-3-methylbutyl-1-formate, and methyl-2-phenylethyl sulphide) as well as a nonpredator-associated fruity odor (n-pentyl acetate) were not avoided. Neither the general motor activity nor the number of excreted fecal pellets, both widely used measures of stress- or anxiety-related behavior, were systematically affected by any of the odorants tested. Further, we found that small changes in the molecular structure of a predator odor component can have a marked effect on its behavioral significance as 2-propylthietane was significantly avoided by the mice whereas the structurally related 2,2-dimethylthietane was not. We conclude that sulfur-containing volatiles identified as characteristic components of the urine, feces, and anal gland secretions of mammalian predators can be, but are not necessarily sufficient to elicit defensive behaviors in a mammalian prey species.

Will Trespassers Be Prosecuted or Assessed According to Their Merits? A Consilient Interpretation of Territoriality in a Group-Living Carnivore, the European Badger (Meles meles).

Socio-spatial interactions of Carnivores have traditionally been described using the vocabulary of territoriality and aggression, with scent marks interpreted as ‘scent fences’. Here, we investigate the role of olfactory signals in assumed territorial marking of group-living solitary foragers using European badgers Meles meles as a model. We presented anal gland secretions (n = 351) from known individuals to identifiable recipients (n = 187), to assess response-variation according to familiarity (own-group, neighbours, strangers) and spatial context (in-context: at a shared border; out-of-context: at an unshared border/ the main sett). Sniffing and over-marking (with subcaudal gland secretion) responses were strongest to anal gland secretions from strangers, intermediate to neighbouring-group and weakest to own-group members. Secretions from both, strangers and neighbours, were sniffed for longer than were own-group samples, although neighbour-secretion presented out-of-context evoked no greater interest than in-context. On an individual level, responses were further moderated by the relevance of individual-specific donor information encoded in the secretion, as it related to the physiological state of the responder. There was a trend bordering on significance for males to sniff for longer than did females, but without sex-related differences in the frequency of subcaudal over-marking responses, and males over-marked oestrous female secretions more than non-oestrous females. There were no age-class related differences in sniff-duration or in over-marking. Evaluating these results in the context of the Familiarity hypothesis, the Threat-level hypothesis, and the Individual advertisement hypothesis evidences that interpretations of territorial scent-marks depicting rigid and potentially agonistic discrimination between own- and foreign-group conspecifics are overly simplistic. We use our findings to advance conceptual understanding of badger socio-spatial ecology, and the general context of territoriality and group-range dynamics.

Sex Discrimination via Anal Gland Secretion in a Territorial Monogamous Mammal

Monogamy in mammals is relatively unusual, only occurring in 3–5% and to an even lesser degree in rodents. Monogamy exists in two forms, facultative which evolved due to female dispersion and obligate when male care is needed for offspring survival. Most monogamous rodents use scent marking as a form of territorial defence to obtain exclusive access to vital resources. Intrasexual competition occurs in many species to maintain pair bonds and also to signal presence within a territory. In this study, we tested the hypothesis that resident obligate monogamous Eurasian beaver (Castor fiber) can discriminate between the sexes when investigating anal gland secretions (AGS) from unknown (stranger) mated pairs. We predicted that, due to intrasexual competition, dominant territorial male and female beavers will display a stronger response to AGS of same-sex conspecifics. Territorial intrusion by mated pair strangers was simulated by the formation of experimental scent mounds (ESM) with AGS. Our results showed that both sexes (1) displayed a stronger response to the male AGS in sniffing duration, (2) physically responded for longer durations towards male AGS and (3) overmarked male AGS more frequently. We infer that obligate monogamous mammals can determine the sexual identity of intruding conspecifics of a mated pair via AGS and that intruding males may pose more of a threat to resident males via intrasexual competition and resident females due to the long-term costs, such as infanticide and abortion, of a new dominant male.

Coexistence Patterns Between Ants And Spiders In Grassland Habitats

The ecological importance of both ants and spiders is well known, as well as the relationship between certain spiders and ants. The two main strategies ˗ myrmecomorphy (ant-mimicking) and myrmecophagy (ant-eating) ˗ that connect spiders to ants have been mostly studied at the behavioural level. However, less is known about how these relationships manifest at the ecological level by shaping the distribution of populations and assemblages. Our question was how ant-mimicking and ant-eating spiders associate with ant genera as revealed by field co-occurrence patterns. For both spider groups we examined strength and specificity of the association, and how it is affected by ant size and defence strategy. To study spider-ant association patterns we carried out pitfall sampling on the dolomitic Sas Hill located in Budapest, Hungary. Spiders and ants were collected at eight grassland locations by operating five pitfalls/location continuously for two years. To find co-occurrence patterns, two approaches were used: correlation analyses to uncover possible spider-ant pairs, and null-model analyses (C-score) to show negative associations. These alternative statistical methods revealed consistent co-occurrence patterns. Associations were generally broad, not specific to exact ant genera. Ant-eating spiders showed a stronger association with ants. Both ant-mimicking and ant-eating spiders associated more strongly with Formicine ants - species with formic acid or anal gland secretions, and had neutral association with Myrmicine ants - species with stings and cuticle defences.

Vomeronasal organ lesion disrupts social odor recognition, behaviors and fitness in golden hamsters.

Most studies support the viewpoint that the vomeronasal organ has a profound effect on conspecific odor recognition, scent marking and mating behavior in the golden hamster (Mesocricetus auratus). However, the role of the vomeronasal organ in social odor recognition, social interaction and fitness is not well understood. Therefore, we conducted a series of behavioral and physiological tests to examine the referred points in golden hamster. We found that male hamsters with vomeronasal organ lesion showed no preference between a predator odor (the anal gland secretion of the Siberian weasels (Mustela sibirica) and putative female pheromone components (myristic acid and palmitic acid), but were still able to discriminate between these 2 kinds of odors. In behavioral tests of anxiety, we found that vomeronasal organ removal causes female hamsters to spend much less time in center grids and to cross fewer center grids and males to make fewer crossings between light and dark boxes than sham-operated controls. This indicates that a chronic vomeronasal organ lesion induced anxious responses in females. In aggressive behavioral tests, we found that a chronic vomeronasal organ lesion decreased agonistic behavior in female hamsters but not in males. The pup growth and litter size show no differences between the 2 groups. All together, our data suggested that vomeronasal organ ablation disrupted the olfactory recognition of social chemosignals in males, and induced anxiety-like and aggressive behavior changes in females. However, a vomeronasal organ lesion did not affect the reproductive capacity and fitness of hamsters. Our studies may have important implications concerning the role of the vomeronasal organ in golden hamsters and also in rodents.