Anal Sac Secretions Research Articles

Sprayed Urine Emits a Pungent Odor due to its Increased Adhesion to Vertical Objects via Urinary Proteins Rather Than to Changes in its Volatile Chemical Profile in Domestic Cats.

Spraying urine on vertical objects by raising the tail is a commonly observed functional behavior for chemical communication in Felidae species, including domestic cats (Felis silvestris catus). The sprayed urine is recognized as a chemical signal for territorial ownership of their habitats. Previous studies reported that sprayed urine emits a more pungent odor than urine excreted from a squatting position. However, little is known about how sprayed urine acts as a strong scent mark in the environment. Here, we showed that sprayed urine originates only from bladder urine without any secretions, such as anal sac secretions, but it can effectively emit volatile organic compounds (VOCs) when smeared on vertical objects due to its strong adhesion. Chemical profiles of VOCs and odor qualities were similar between fresh sprayed urine and bladder urine sampled immediately after spraying from the same individuals. Meanwhile, feline-specific proteinuria arising from excretion of a carboxylesterase that produces a precursor of cat-specific odorants resulted in reduced surface tension of the urine and increased adhesion to vertical surfaces, which kept sprayed urine on the surfaces and led to the emission of large amounts of VOCs. In conclusion, proteinuria contributes to the emission of a strong odor through its enhanced adhesion to vertical objects without other secretions containing malodorous substances. These findings improve our understanding of the mechanism of scent marking via the spraying of urine for chemical communication in cats.

Bacterial microbiota and proinflammatory cytokines in the anal sacs of treated and untreated atopic dogs: Comparison with a healthy control group.

The pathogenesis of anal sacculitis has not been extensively investigated, although atopic dogs seem to be predisposed to the disease. The aim of this study was therefore to characterize and compare the bacterial microbiota and pro-inflammatory cytokines in the anal sacs of dogs from three groups (healthy dogs, untreated atopic dogs and atopic dogs receiving antipruritic treatment or allergen-specific immunotherapy) in order to determine whether changes could be at the origin of anal sacculitis in atopic dogs. Bacterial populations of anal sac secretions from fifteen healthy dogs, fourteen untreated and six treated atopic dogs were characterized by sequencing the V4 region of the 16S rRNA gene using Illumina technology. Proinflammatory cytokines were analyzed with the Luminex multiplex test. Community membership and structure were significantly different between the anal sacs of healthy and untreated atopic dogs (P = 0.002 and P = 0.003, respectively) and between those of untreated and treated atopic dogs (P = 0.012 and P = 0.017, respectively). However, the community structure was similar in healthy and treated atopic dogs (P = 0.332). Among the proinflammatory cytokines assessed, there was no significant difference between groups, except for interleukin 8 which was higher in the anal sacs of untreated atopic dogs compared to treated atopic dogs (P = 0.02), and tumor necrosis factor-alpha which was lower in the anal sacs of healthy dogs compared to treated atopic dogs (P = 0.04). These results reveal a dysbiosis in the anal sacs of atopic dogs, which may partially explain the predisposition of atopic dogs to develop bacterial anal sacculitis. Treatments received by atopic dogs (oclacitinib, desloratadine and allergen-specific immunotherapy) shift the microbiota of the anal sacs towards that of healthy dogs. Further studies are required to identify significant cytokines contributing to anal sacculitis in atopic dogs.

Comparison of volatile compounds of anal sac secretions between the sexes of domestic dog (Canis lupus familiaris).

Volatile compounds of anal sac secretions are odorant chemicals used across the carnivores for social communication such as identifying individuals and group membership. Odor profiles taken from expressed anal sac secretions of some species of carnivores have been detected in previous studies. In this study, the volatile compounds of anal sac secretions between five male and five female domestic dogs (Canis lupus familiaris) were compared. Volatile chemicals were extracted, separated, and analyzed by gas chromatography-mass spectrometry with solid-phase micro-extraction and identified from their electron ionization mass spectra and Kovats retention indices. The results showed the presence of various types of compounds including organic fatty acids, ketones, aldehydes, esters, and alcohols in the anal sac secretions of dogs. Greater amounts of diversity and esters, and lower amounts of alcohols were detected in the anal sac secretions of females compared to males. This was accompanied by finding citrate and acetic acid ester only in the females. Furthermore, presence of some sex-specific organic compounds like dimethylcyclopentyl ethanone indicates that the volatile profiles of anal sac secretions in 10 domestic dogs are differentiated by host sex.

Bacteria isolated from Bengal cat (Felis catus × Prionailurus bengalensis) anal sac secretions produce volatile compounds potentially associated with animal signaling.

In social animals, scent secretions and marking behaviors play critical roles in communication, including intraspecific signals, such as identifying individuals and group membership, as well as interspecific signaling. Anal sacs are an important odor producing organ found across the carnivorans (species in the mammalian Order Carnivora). Secretions from the anal sac may be used as chemical signals by animals for behaviors ranging from defense to species recognition to signaling reproductive status. In addition, a recent study suggests that domestic cats utilize short-chain free fatty acids in anal sac secretions for individual recognition. The fermentation hypothesis is the idea that symbiotic microorganisms living in association with animals contribute to odor profiles used in chemical communication and that variation in these chemical signals reflects variation in the microbial community. Here we examine the fermentation hypothesis by characterizing volatile organic compounds (VOC) and bacteria isolated from anal sac secretions collected from a male Bengal cat (Felis catus × Prionailurus bengalensis), a cross between the domestic cat and the leopard cat. Both left and right anal sacs of a male Bengal cat were manually expressed (emptied) and collected. Half of the material was used to culture bacteria or to extract bacterial DNA and the other half was used for VOC analysis. DNA was extracted from the anal sac secretions and used for a 16S rRNA gene PCR amplification and sequencing based characterization of the microbial community. Additionally, some of the material was plated out in order to isolate bacterial colonies. Three taxa (Bacteroides fragilis, Tessaracoccus, and Finegoldia magna) were relatively abundant in the 16S rRNA gene sequence data and also isolated by culturing. Using Solid Phase Microextraction (SPME) gas chromatography-mass spectrometry (GC-MS), we tentatively identified 52 compounds from the Bengal cat anal sac secretions and 67 compounds from cultures of the three bacterial isolates chosen for further analysis. Among 67 compounds tentatively identified from bacterial isolates, 51 were also found in the anal sac secretion. We show that the bacterial community in the anal sac consists primarily of only a few abundant taxa and that isolates of these taxa produce numerous volatiles that are found in the combined anal sac volatile profile. Several of these volatiles are found in anal sac secretions from other carnivorans, and are also associated with known bacterial biosynthesis pathways. This is consistent with the fermentation hypothesis and the possibility that the anal sac is maintained at least in part to house bacteria that produce volatiles for the host.

Identification of putative volatile sex pheromones in female domestic dogs (Canis familiaris)

More than hundred urine samples, vaginal secretions, vulval and anal imprints as well as anal sac secretions, collected during various phases of the ovarian cycle, were evaluated with the HS-SPME/GC–MS technique. The results indicate there were differences among samples of urine collected during particular phases of the ovarian cycle. Unequivocally, the amount of carbonyl aromatic compounds, such as acetophenone (hypnone) and benzaldehyde, increased during the period of proestrus and estrus of the estrous cycle. This was accompanied by increases in methylketones, e.g., 2-octanone, 2-pentanone and 3-hexanone. Simultaneously, amounts of sulfide compounds (1-methylotiopropane; 1-methylotiobutane, 1-methylotipentane and dimethyl trisulfide) decreased during the period of estrus and abruptly increased in diestrus. These observations suggest a possible dual mechanism of interaction between males and females during and subsequent to the mating period, including the existence of both attracting and repelling signals. No significant changes were detected in samples other than urine. Further studies including a proteomic approach as well as behavioral assessments are suggested to identify the complete range of estrual semiochemical signals and to verify the semiochemical activities of identified substances.

Identification of 2-phenylethanol with a rose-like odor from anal sac secretions of the small Indian mongoose (Herpestes auropunctatus).

The small Indian mongoose (Herpestes auropunctatus) is an invasive species in Okinawa and Amami-Oshima, Japan. Major strategies for their eradication have been the use of baited traps, which suffer from decreasing efficiency with declining populations and the bycatch of native animals. To address these concerns, mongoose-specific lures are required. In this study, we aimed to identify species- and/or sex-specific compounds from anal sac secretions of small Indian mongooses. Volatile compounds emitted from male and female mongoose anal sac secretions were analyzed by thermal desorption-gas chromatography-mass spectrometry. In addition to several fatty acids, 2-phenylethanol was identified as a minor compound, which is uncommon in mammalian secretions but a dominant odorant in roses. Female samples emitted higher levels of 2-phenylethanol than male samples did. These findings indicate that 2-phenylethanol is a female-specific volatile compound of anal sac secretions in small Indian mongooses, and it may be useful as an ingredient of mongoose-specific scent lures.

Olfactory discrimination of anal sac secretions in the domestic cat and the chemical profiles of the volatile compounds

Scent emitted from anal sac secretions provides important signals for most Carnivora. Their secretions emit a variety of volatile compounds, some of which function as chemical signals with information about the scent owners. The domestic cat has a pair of anal sac glands to secrete a pungent liquid. Their anal sac secretions may give information about sex, reproductive state, and recognition of individuals. However, little is known about the volatile compounds emitted from anal sac secretions and their biological functions in cats. In this study, we examined the volatile chemical profiles of anal sac secretions in cats and their olfactory ability to discriminate intraspecific anal sac secretions. Analysis with gas chromatography–mass spectrometry showed that the major volatile compounds were short-chain free fatty acids, whose contents varied among individuals, as well as other carnivores. There was no sex difference in the volatile profiles. In temporal analyses of individual anal sac secretions performed 2 months apart, the profiles were highly conserved within individuals. Habituation–dishabituation tests showed that cats can distinguish individual differences in the odor of anal sac secretions. These results suggest that cats utilize short-chain free fatty acids emitted from anal sac secretions to obtain scent information for individual recognition rather than species or sex recognition.

Draft Genome Sequence of Propionibacterium avidum Strain UCD-PD2 Isolated from a Feline Anal Sac.

ABSTRACTHere, we present the draft genome sequence of Propionibacterium (Cutibacterium) avidum strain UCD-PD2. The assembly contains 2,667,287 bp in 51 contigs. The strain was isolated from anal sac secretion samples collected from a feral domestic cat (Felis catus) as part of a larger project to study the microbiology of cats.

Draft Genome Sequence of Enterococcus faecalis Strain UCD-PD3.

Here, we present the draft genome sequence of Enterococcus faecalis strain UCD-PD3. The assembly contains 2,861,314 bp in 73 contigs. This strain was isolated from a feral domestic cat (Felis catus) anal sac secretion sample, as part of a project on isolating and characterizing the microbes present in feline anal sacs.

River otter population size estimation using noninvasive latrine surveys

AbstractAcross much of North America, river otter (Lontra canadensis) populations were extirpated or greatly reduced by the early 20th century. More recently, reintroductions have resulted in restored populations and the recommencement of managed trapping. Perhaps the best example of these river otter reintroductions occurred in Missouri, regarded as one of the most successful carnivore recovery programs in history. However, abundance estimates for river otter populations are difficult to obtain and often contentious when used to underpin management activities. We assessed the value of latrine site monitoring as a mechanism for quantifying river otter abundance. Analyses of fecal DNA to identify individual animals may result in an improved population estimate and have been used for a variety of mammal species. We optimized laboratory protocols, redesigned existing microsatellite primers, and calculated genotyping error rates to enhance genotyping success for a large quantity of river otter scat samples. We also developed a method for molecular sexing. We then extracted DNA from 1,421 scat samples and anal sac secretions (anal jelly) collected during latrine site counts along 22–34‐km stretches representing 8–77% of 8 rivers in southern Missouri in 2009. Error rates were low for the redesigned microsatellites. We obtained genotypes at 7–10 microsatellite loci for 24% of samples, observing highest success for anal jelly samples (71%) and lowest for fresh samples (collected within 1 day of defecation). We identified 63 otters (41 M, 22 F) in the 8 rivers, ranging from 2 to 14 otters per river. Analyses using program CAPWIRE resulted in population estimates similar to the minimum genotyping estimate. Density estimates averaged 0.24 otters/km. We used linear regression to develop and contrast models predicting population size based on latrine site and scat count indices, which are easily collected in the field. Population size was best predicted by a combination of scats per latrine and latrines per kilometer. Our results provide methodological approaches to guide wildlife managers seeking to initiate similar river otter fecal genotyping studies, as well as to estimate and monitor river otter population sizes. © 2011 The Wildlife Society.

Chemical scent constituents in feces of wild Iberian wolves (Canis lupus signatus)

In spite of the importance of feces in scent marking and intraspecific chemical communication of many mammals, only a few studies have examined the chemical constituents of feces that could have a signaling function. We described here the chemicals found in feces recently deposited in the wild by adult and pup Iberian wolves (Canis lupus signatus). By means of analyses using gas chromatography–mass spectrometry (GC–MS), we identified 77 compounds in feces of adult wolves, mainly heterocyclic aromatic organic compounds, such as indole or phenol, but also steroids, such as cholesterol, carboxylic acids and their esters between n-C3 and n-C18, aldehydes, alcohols and significant quantities of squalene and α-tocopherol, which would increase the chemical stability of feces on humid substrates. Some of the odoriferous compounds are very likely originated in the anal-sac secretions, and later incorporated onto feces during defecation. However, the abundance of the main odoriferous compounds in feces could be explained by the result of the bacterial action on food components. Feces of pups only had 26 compounds, most of these were also found in adults’ feces, but four compounds were only found in pups’ feces, and there were also age-related differences respect to the proportions of shared chemicals. These differences could be partly due to the absence of anal-sac glands in pups, which would not need to scent mark substrates.

Comparison of anal sac cytological findings and behaviour in clinically normal dogs and those affected with anal sac disease

No previous study has explored the relationship between cytology and the frequency of behaviours associated with anal sac disease (ASD). The goals of the study were: (i) to compare the cytological findings between anal sac secretions from normal dogs with no history of ASD to those with non-neoplastic ASD; (ii) to determine whether anal sac cytological findings can be used to differentiate between normal dogs and dogs with ASD; (iii) to explore the correlation of anal sac cytology and behaviour between normal dogs and dogs with ASD; and (iv) to describe behaviours typical of ASD as reported by owners. Thirty dogs were selected for this study, based on their behavioural history as detailed in a questionnaire completed by their owners. Of the thirty dogs, ten were considered normal insofar as they had no history of ASD clinical signs. The remaining 20 dogs were characterized as having ASD, with a chronic history of perianal pruritus, but no other pruritus. All dogs had their anal sacs manually expressed, and the discharge was examined microscopically in a blinded manner. A total of 171 oil immersion fields (OIFs) were examined from normal dogs and 333 OIFs from dogs with ASD. The behavioural results for dogs with ASD revealed that scooting recurred with a median frequency of 3 weeks post-anal sac expression. There were no clinically statistically significant cytological differences between normal dogs and those with ASD, thereby leading to the conclusion that cytology is an ineffective tool for diagnosing ASD.

Induced Estrus in Female Small Asian Mongooses (Herpestes javanicus) for the Purpose of Controlling Invasive Alien Species in Okinawa Island

The small Asian mongoose (Herpestes javanicus) belongs to the family Herpestidae, order Carnivora, and originally inhabited Iraq through northern India to extreme southern China and Indonesia, the Malay Peninsula, Hainan, Java (Nowak 2005). Mongooses, including small Asian mongooses, have been widely introduced by people to exterminate rats for pest control (Yamada and Sugimura 2004). In Japan, small Asian mongooses were introduced to eliminate yellow-spotted pit vipers (Habu snakes; Protobothrops flavoviridis) and sugarcane rats on Okinawa Island in 1910 and Amami-Oshima Island, Kagoshima Prefecture in 1979 (Yamada 2002). Naturalized and overbred mongooses, which prey on insects, reptiles, small mammals, wild birds and domestic fowl, have impacted the poultry industry, and threaten the survival of rare endemic species, such as the Amami rabbit (Pentalagus furnessi), Okinawa rail (Galirallus okinawae) and so on (Yamada 2002). A mongoose eradication project led by the Ministry of the Environment, Government of Japan, has been conducted on Amami-Oshima Island since 1996 and northern Okinawa Island since 2000. Moreover, mongooses were designated as invasive alien species by the Invasive Alien Species Act in Japan (Ministry of the Environment, 2004); however, the eradication project has not yet succeeded. The distribution of mongooses has expanded on Okinawa and Amami-Oshima Islands and, in addition, mongooses were found in Kagoshima city (Kyusyu Island) in 2007. Currently, cage-type traps and cylindrical trapping with bait, chicken meat, have been mainly used for capturing mongooses on Okinawa and Amami-Oshima Islands. Mongoose-drawing techniques require a more effective for increase of the capture number. For effective capture, studies searching for and identifying attractant substances for species-specific capture have been performed in small Asian mongooses, and the previous study attempted to identify volatile fatty acids from anal sac secretions in this species (Ogura et al. 2000b). In the present study, to establish a trapping method with an estrous female or its excreta, we tried to create a protocol to artificially induce estrus in female small Asian mongooses using exogenous hormonal treatment. Estrus was determined based on the profile of urinary estrogen concentrations.

Gross and cytological characteristics of normal feline anal-sac secretions.

This study was conducted to characterize the gross and cytological characteristics of secretions from normal feline anal sacs. Thirty cats with no recent history of anal-sac disease were selected consecutively and their anal sacs manually expressed. The color, cell counts, or presence of solid portions of the secretions in the youngest cats (<1 year old) were not significantly different from the rest. Young cats (<1 year old) tended to more often have watery secretions. The secretions contained 13 (median) epithelial cells per 600x magnification microscopic field, basophilic background debris, and a mixture of Gram-positive cocci (83 per field; median), Gram-negative cocci (38 per field; median), and Gram-negative rods (1.8 per field; median). Most secretions had occasional neutrophils, and a minority of cats had occasional yeasts in their secretion. Erythrocytes were found only rarely. Extreme heterogeneity existed in gross color, consistency, and amount of solid material. Hence, these latter parameters would not be valid indicators of anal-sac disease.

A computational investigation of sulfur-containing heterocyclic components from the anal sac secretions of Mustela species

A computational investigation of the sulfur-containing heterocyclic components (substituted thietanes and 1,2-dithiolanes) of Mustela anal sac secretions has been carried out. A cluster analysis of the chemical compositions of Mustela anal sac volatiles reveals little similarity with established phylogenetic relationships between members of the genus. Ab initio calculations [MP2/6-311++G(2df,2p)//B3LYP/6-311++G**] show the lowest-energy C5H10S isomeric thietane to be 2,2-dimethylthietane, which is also the most abundant of the Mustela thietanes. Similarly, 3,3-dimethyl-1,2-dithiolane is the lowest-energy C5H10S2 compound. 2-n-Propylthietane is the highest-energy C6H12S compound, but the most abundant Mustela C6H12S compound produced, whereas cis-2-ethyl-4-methylthietane, the lowest-energy C6H12S thietane, has never been observed in Mustela anal sac secretions. A molecular docking analysis of the Mustela sulfur-containing heterocycles into both porcine and bovine odorant binding proteins reveals the interactions of the docked ligands with the proteins to be largely hydrophobic, and have binding energies generally lower than typical odorant molecules such as linalool or eugenol.

Gross and cytological characteristics of normal canine anal-sac secretions.

A study was conducted to characterize the gross and cytological characteristics of secretions from normal canine anal sacs. Fifty dogs with no recent history of anal-sac disease were selected consecutively and their anal sacs manually expressed. Of the 47 dogs with known or estimated ages, 27 were < 1 year olds. However, these youngest dogs were not significantly different from the rest in cell counts, viscosity, or presence of dark or light solid portions of the secretions. For all dogs combined, the secretions typically contained 41 (median) corneocytes per 400x magnification microscopic field, lots of basophilic background debris, and a mixture of Gram-positive cocci (173 per field; median), Gram-positive rods (29 per field; median), and Gram-negative rods (two per field; median). Although median counts across 10 400x fields per sac indicated no leucocytes or yeasts, many of the normal dogs had an occasional 400x field with non-degenerate neutrophils and yeasts. Erythrocytes were not a feature of these anal-sac secretions, and mononuclear leucocytes were found only rarely. Extreme heterogeneity existed in gross colour, consistency and the amount of solid material. Thus, these latter parameters probably would not be valid indicators of anal-sac disease.

Volatile components in defensive spray of the hooded skunk, Mephitis macroura.

GC-MS analysis of the anal sac secretion from the hooded skunk, Mephitis macroura, showed the following seven major components comprised 99% of the volatiles in this secretion: (E)-2-butene-1-thiol, 3-methyl-1-butanethiol, S-(E)-2-butenyl thioacetate, S-3-methylbutenyl thioacetate, 2-phenylethanethiol, 2-methylquinoline, and 2-quinolinemethanethiol. Minor volatile components identified in this secretion are phenylmethanethiol, S-phenylmethyl thioacetate. S-2-phenylethyl thioacetate, bis[(E)-2-butenyl] disulfide, (E)-2-butenyl 3-methylbutyl disulfide, bis(3-methylbutyl) disulfide, and S-2-quinolinemethyl thioacetate. This secretion is similar to that of the striped skunk, Mephitis mephitis, differing only in that it contains four compounds not reported from the striped skunk: phenylmethanethiol, S-phenylmethyl thioacetate, 2-phenylethanethiol, and S-2-phenylethyl thioacetate.

Responses to olfactory stimuli in spotted hyenas (Crocuta crocuta): II. Discrimination of conspecific scent.

Scent marking in spotted hyenas (Crocuta crocuta) includes the deposition of anal sac secretions, or "paste," and presumably advertises territorial ownership. To test whether captive hyenas classify and discriminate individuals using odor cues in paste, the authors conducted behavioral discrimination bioassays and recorded hyena investigation of paste extracted from various conspecific donors. In Experiment 1, subjects directed most investigative behavior toward scents from unfamiliar hyenas and members of the opposite sex. In Experiment 2, male hyenas discriminated between concurrent presentations of paste from various unfamiliar females in similar reproductive states. Thus, pasted scent marks convey information about the sex, familiarity, and even identity of conspecifics. Aside from territory maintenance, scent marking may also communicate information about individual sexual status.

The utility of chemical signals as phylogenetic characters: an example from the Felidae

Chemical secretions that are explicitly tied to species recognition may potentially be informative for phylogenetic reconstruction, especially when traditional morphological or molecular characters lack resolution. Anal sac secretions from 16 species within the family Felidae (order Carnivora) were chemically analysed and their utility as phylogenetic characters was assessed. Results were generally consistent across the different chemical data types (e.g. glycolipids, neutral lipids, or phospholipids). Two major clades were indicated, falling out according to body size: one for species greater than 30 kg (Panthera, Uncia, and Puma) and another for those less than 12 kg (remaining species). The primary solutions agreed with respect to the species pairs Prionailurus+Leptailurus, Caracal+Lynx,Oncifelis+Leopardus , Otocolobus+Felis, Panthera leo+P. pardus, and P. tigris+Uncia. The only area of disagreement between chemical types was the positioning of the mountain lion (Puma concolor); however, this species appears to cluster with the cheetah (Acinonyx jubatus) in the «big cat» clade. Although our solutions differ from six previously proposed hypotheses of felid phylogeny (morphological and molecular), the previous estimates all differ strongly amongst themselves reflecting the historical uncertainty regarding felid systematics. Phylogenies derived from the lipid data were very robust and decisive. Few equally most parsimonious trees were obtained, consistency indices were much higher than their expected values, and bootstrap and Bremer support values were also high. Thus, our findings illustrate the species-specific nature of chemical signals and their usefulness as phylogenetic characters.

The utility of chemical signals as phylogenetic characters: an example from the Felidae

Abstract Chemical secretions that are explicitly tied to species recognition may potentially be informative for phylogenetic reconstruction, especially when traditional morphological or molecular characters lack resolution. Anal sac secretions from 16 species within the family Felidae (order Carnivora) were chemically analysed and their utility as phylogenetic characters was assessed. Results were generally consistent across the different chemical data types (e.g. glycolipids, neutral lipids, or phospholipids). Two major clades were indicated, falling out according to body size: one for species greater than 30 kg ( Panthera , Uncia , and Puma ) and another for those less than 12 kg (remaining species). The primary solutions agreed with respect to the species pairs Prionailurus+Leptailurus , Caracal+Lynx , Oncifelis+Leopardus , Otocolobus+Felis , Panthera leo+P. pardus , and P. tigris+Uncia . The only area of disagreement between chemical types was the positioning of the mountain lion ( Puma concolor ); however, this species appears to cluster with the cheetah ( Acinonyx jubatus ) in the «big cat» clade. Although our solutions differ from six previously proposed hypotheses of felid phylogeny (morphological and molecular), the previous estimates all differ strongly amongst themselves reflecting the historical uncertainty regarding felid systematics. Phylogenies derived from the lipid data were very robust and decisive. Few equally most parsimonious trees were obtained, consistency indices were much higher than their expected values, and bootstrap and Bremer support values were also high. Thus, our findings illustrate the species-specific nature of chemical signals and their usefulness as phylogenetic characters.