Molecular systematics of new world gopher, bull, and pinesnakes (Pituophis: Colubridae), a transcontinental species complex

Pituophis melanoleucus (gopher, bull, and pinesnakes) is among the most widely distributed polytypic species complexes in North America, with most authors recognizing from a single transcontinental species (the melanoleucus complex, composed of 15 subspecies) to four (monotypic and polytypic) species. We used mitochondrial gene sequences from the two middle American species, P. deppei and P. lineaticollis, and from 13 subspecies from most of the range of the melanoleucus complex to test various phylogenetic hypotheses for Pituophis. Maximum parsimony and maximum likelihood methods identified the same major clades within Pituophis and indicated that two segments of the melanoleucus complex, the lodingi-melanoleucus-mugitus eastern pinesnake clade and the affinis-annectens-bimaris-catenifer-deserticola- sayi-ruthveni-vertebr alis clade from central and western United States and northern Mexico, represent divergent, allopatric lineages with no known intergradation zone. We recognize each of these two groupings as a different species. Our data also indicate that some ruthveni are more closely related to sayi than to other ruthveni. Nonetheless, ruthveni is an allopatric taxon diagnosable from its closest relatives by a combination of morphometric characters, and because it is likely that at least some of these traits are independent and genetically inherited, we interpret this as evidence that ruthveni has attained the status of independent evolutionary lineage, despite the fact that it retains strong genetic affinities with sayi. The endemic Baja Californian gopher snakes (bimaris and vertebralis) are considered by some taxonomists as a different species, P. vertebralis, but we discovered that these serpents belong to two different clades and hence we do not agree with the recognition of P. vertebralis as presently defined. In summary, we believe that three distinct species are included in the melanoleucus complex, Pituophis melanoleucus (sensu stricto), P. catenifer, and P. ruthveni, and that their recognition better represents the evolutionary diversity within this species complex.     

Optimal patch use by foraging workers of Lasius fuliginosus, L. niger and Myrmica ruginodis

1. The predictions of the marginal value theorem, that foragers should spend more time in both better quality patches and more distant patches, were tested with three European ant species, Lasius fuliginosus , L. niger and Myrmica ruginodis.
2. As the quality of patches of sucrose solution increased, the feeding time of foraging workers of L. niger also increased.
3. At constant patch quality, feeding times of L. niger and M. ruginodis increased with increasing distance of the patch from the nest entrance.
4. Foraging workers of L. fuliginosus showed a similar response to patch distance in the field, but feeding times were also significantly influenced by air temperature, decreasing as temperature increased.
5. These results show qualitative agreement with the predictions of the marginal value theorem.     

Intraspecific variation in advertisement call characteristics and acoustic strategies among male forest green tree frogs

Frogs are a representative taxon that use advertisement calls to aid in reproduction. In most frog species, calls vary with body size, and allometric constraints between body size and call frequency have been widely reported among anuran species. Although this variation is an important driver of sexual selection in frogs, male advertisement call strategies may also vary according to body size. In this study, we conducted playback experiments on the male forest green tree frog (Zhangixalus arboreus) to determine whether male advertisement call characteristics and strategies vary according to body size and the amplitude of intraspecific chorus noise. The results indicated that the calls of larger individuals are louder and lower than those of smaller ones, who call more frequently; moreover, the calls become lower, and the number of calls decreases, as noise levels increase. These findings suggest that forest green tree frog emits lower calls or refrains from calling when chorus noise increases, and that intraspecific variation in advertisement call characteristics can induce different strategies in response to chorus noise. Because advertisement call variation with body size is common among frog species, intraspecific variation in male advertisement call strategies may also be a common phenomenon.     

Pacific treefrogs use temporal integration to differentiate advertisement from encounter calls

Male Pacific treefrogs, Hyla regilla, use advertisement and encounter calls in their reproductive communication. Encounter calls are produced when a male hears either advertisement or encounter calls at amplitudes that exceed his aggressive threshold for that call type, and they are believed to play a role in male spacing in a chorus. Aggressive thresholds of males for advertisement and encounter calls are plastic; males resume advertisement calling (i.e. accommodate) after repeated exposure to either advertisement or encounter calls that exceed their aggressive thresholds for that call type. Presentation of one call type does not elevate a male's aggressive threshold for the other call type, indicating that discrete neural ‘channels’ exist for processing advertisement versus encounter calls. Because these calls are spectrally highly similar, it is likely that they are differentiated in the central auditory system on the basis of their different temporal structure; pulses are repeated in advertisement and encounter calls at rates of approximately 100 pulses/s and 30 pulses/s, respectively. The present study tested whether consecutive advertisement-type interpulse intervals (IPIs) are required to activate the filter for advertisement calls. Males were accommodated to synthetic stimuli that differed in the proportion of advertisement versus encounter IPIs. Aggressive thresholds of males for the encounter call and advertisement call (monophasic type) were elevated 450% and 58%, respectively, following accommodation to a stimulus that had nine of each IPI type, arranged in an alternating sequence. Aggressive thresholds for the monophasic advertisement call were elevated 151%, however, when males were accommodated with stimuli that had two sets of eight to nine consecutive advertisement IPIs; consistent with previous work, aggressive thresholds for the encounter call were not significantly elevated. Aggressive thresholds for the advertisement call are typically elevated approximately four-fold following accommodation with that call type. These findings are consistent, therefore, with the hypothesis that several consecutive advertisement IPIs are required to activate the ‘advertisement channel’; if consecutive advertisement IPIs were not required, the stimulus that consisted of the same number of advertisement and encounter IPIs should have induced comparable elevations of the aggressive thresholds of males for advertisement and encounter calls. Selective filters for the advertisement call, therefore, appear to integrate information across many consecutive ‘correct’ IPIs. This integration process may contribute to a female's call preference.Copyright 2002 The Association for the Study of Animal Behaviour. Published by Elsevier Science Ltd. All rights reserved .     

Differentiation of vocalizations in bushbabies (Galaginae, Prosimiae, Primates) and the significance for assessing phylogenetic relationships

A comparative bioacustic analysis of vocalizations of the prosimian subfamily Galaginae revealed that morphologically similar sibling taxa within the main groups of the lesser galagos and the greater galagos can be reliably identified phenotypically on the basis of the acoustic structure of their loud call or advertisement call. Results confirm the separation of two distinct species of greater galagos, Galago crassicaudatus and Galago garnettii, and strongly suggest the discrimination of three distinct species from the senegalensis lesser bushbaby group, Galago senegalensis, Galago moholi and Galao zanzibaricus. An investigation of the ontogenetic development of the loud call indicated that it is derived from the infant's isolation call, displaying in all studied bushbaby taxa a fairly similar acoustic pattern. Shared acoustic characters of the loud call among the different taxa as well as the infant's isolation call were used to propose a hypothesis about the phylogenetic affinities in bushbabies. The results seem to be supported by recent fossil records.     

Genetic population structure and call variation in a passerine bird, the satin bowerbird, Ptilonorhynchus violaceus

Geographic variation in vocalizations is widespread in passerine birds, but its origins and maintenance remain unclear. One hypothesis to explain this variation is that it is associated with geographic isolation among populations and therefore should follow a vicariant pattern similar to that typically found in neutral genetic markers. Alternatively, if environmental selection strongly influences vocalizations, then genetic divergence and vocal divergence may be disassociated. This study compared genetic divergence derived from 11 microsatellite markers with a metric of phenotypic divergence derived from male bower advertisement calls. Data were obtained from 16 populations throughout the entire distribution of the satin bowerbird, an Australian wet-forest-restricted passerine. There was no relationship between call divergence and genetic divergence, similar to most other studies on birds with learned vocalizations. Genetic divergence followed a vicariant model of evolution, with the differentiation of isolated populations and isolation-by-distance among continuous populations. Previous work on Ptilonorhynchus violaceus has shown that advertisement call structure is strongly influenced by the acoustic environment of different habitats. Divergence in vocalizations among genetically related populations in different habitats indicates that satin bowerbirds match their vocalizations to the environment in which they live, despite the homogenizing influence of gene flow. In combination with convergence of vocalizations among genetically divergent populations occurring in the same habitat, this shows the overriding importance that habitat-related selection can have on the establishment and maintenance of variation in vocalizations.     

Acoustically induced call modification in the white-lipped frog,Leptodactylus albilabris

The vocalization behaviour of Leptodactylus albilabris was investigated using field playback experiments. To assess the response of males to pre-recorded natural ‘chirp’ (advertisement call) and natural ‘chuckle” (aggressive call) stimuli of gradually increasing broadcast intensity, three parameters (intensity, dominant frequency and repetition rate) of the chirp call were analysed. Of the males tested, 69% showed a significant increase in chirp intensity with increased levels of both stimulus types. In response to playback of the chirp stimulus, males actively modified the dominant frequency of their chirp calls over a mean range of 91·42 Hz, and in one case as much as 400 Hz. Moreover, 12 of 17 males shifted the frequency of their call towards the dominant frequency of the chirp stimulus (2175 Hz) by either increasing or decreasing the dominant frequency of their chirp calls. In response to the natural chuckle stimulus, 83% of the males showed either a decrease or no significant change in the dominant frequency of their chirps. All eight males for which both the chirp frequency and intensity were analysed and that showed an increase in chirp intensity also showed a concomitant increase in chirp dominant frequency. These results are the first to document quantitatively the plasticity of advertisement call intensity and dominant frequency in an anuran. The possible effects of advertisement call modification on male mating success in L. albilabris is discussed.     

Patterns of variability are consistent across signal types in the treefrog Dendropsophus ebraccatus

Animal signal characteristics vary at multiple levels, and this variation can be related to the selective forces acting on signal structure. However, the effectiveness of selection acting on any one signal type may depend on selective forces acting on the same characteristics in other signal types, particularly if the signals share a common physical production mechanism. In anurans, signal variability has been related to the potential for various forms of selection by female choice on the characteristics of male advertisement calls. The significance of variability in the characteristics of another vocalization type, the aggressive call, is less well understood. In the present stiudy, within‐ and between‐male variability was measured in several characteristics of both advertisement and aggressive calls in the treefrog Dendropsophus ebraccatus. Many characteristics of both call types were repeatable within males. Fine‐temporal and spectral characteristics generally had low variation within males, whereas gross‐temporal characteristics were more variable. Unexpectedly, there were strong correlations between both measures of variability and mean values of the call characteristics of advertisement and aggressive calls. Thus, despite potentially conflicting selection pressures acting on advertisement and aggressive calls, the characteristics of individuals' calls were consistent across these two signal types. The potential forces responsible for consistency across call types are discussed, including morphological constraints and behavioural syndromes that act across different signalling contexts. In general, it is argued that measurements of variability made across multiple signal types, as in the present study, can provide important insights into the evolution of signal structure in organisms with complex signal repertoires. © 2013 The Linnean Society of London, Biological Journal of the Linnean Society, 2013, 109 , 131–145.     

Behavioral thermoregulation and the role of melatonin in a nocturnal snake

Daily and seasonal variations in hormone levels influence the complex interactions between behavior and physiology. Ectothermic animals possess the unique ability behaviorally to adjust body temperature (T(b)) to control physiological rate processes. Thus, a hormone may indirectly influence a physiological rate by directly influencing the behaviors that adjust or control that rate process. Although many hormonal influences on behavioral regulation of T(b) remain uninvestigated, melatonin (MEL) generally is considered a hormone that decreases mean preferred T(b). Many ectotherms demonstrate the selection of lower T(b)'s in response to increased MEL concentrations. Here, we examined the influence of MEL on the behavioral regulation of T(b) in the nocturnal African house snake Lamprophis fuliginosus. A series of experiments with two injection regimes of MEL had no significant effect on the mean preferred T(b) of L. fuliginosus. In addition, mean preferred T(b)'s during the photophase did not differ significantly from those during scotophase. Our findings suggest that L. fuliginosus does not respond to elevated concentrations of either endogenous or exogenous MEL. To verify that the African house snake is nocturnal, we investigated activity patterns of L. fuliginosus throughout the photoperiod. The activity period of L. fuliginosus occurs in the scotophase of the photoperiod, a pattern consistent with that of nocturnal species. This suggests that nocturnal organisms such as L. fuliginosus may not respond to MEL in the same manner as many diurnal species. Our results support the hypothesis that some animals, particularly nocturnal species, may have developed alternative responses to increased plasma concentrations of MEL.     

ALLOZYME AND ADVERTISEMENT CALL VARIATION IN THE TÚNGARA FROG,PHYSALAEMUS PUSTULOSUS

We analyzed variation in advertisement calls and allozymes in 30 populations along a 5000-km transect throughout most of the range of the túngara frog, Physalaemus pustulosus. All 12 call variables measured show significant differences among populations despite the importance of the advertisement call in species recognition. Some call variables exhibited clinal variation, whereas most others differed between the two major allozyme groups that have invaded Panama at different times, perhaps 4–4.5 million yr apart. Call variables that primarily affect discrimination among conspecifics tended to exhibit greater variation than call variables that are crucial for species recognition. The proximate mechanism of production underlying a call variable, however, is a better predictor of its variation. Contrary to predictions of some sexual selection models, call variation exhibits predictable patterns of geographical variation, although a substantial portion of variation among populations is not explained by geographic position. Although allozymes, calls, and geography usually covary, closer populations can have more similar calls independent of allozyme similarity.     

Vasotocin maintains multiple call types in the gray treefrog, Hyla versicolor.

The neuropeptide arginine vasotocin (AVT) influences vocalizations in some anuran amphibians but it is unknown whether AVT alters all vocal behaviors of a species similarly. We first characterized the vocal repertoire of male gray treefrogs (Hyla versicolor). Three different call types were distinguished by unique sets of temporal and spectral features. Second, we examined the effects of AVT on each call type by injecting frogs with either AVT (100 microg; intraperitoneal) or saline and recording subsequent behavior. In the field, AVT maintained advertisement calling, whereas calling ceased in saline-injected animals. Advertisement call rate in AVT-injected males fell significantly and dominant frequency of the call was significantly higher. In the laboratory, AVT induced advertisement calling in males that were not initially vocalizing and dominant frequency was also significantly higher in these males. AVT maintained aggressive calling similarly but the characteristics of aggressive calls were not altered by AVT. There were no significant differences in release call behavior between AVT- and saline-injected groups; however, release call duration decreased significantly in AVT-injected animals, compared with preinjection values for the same animals. The effects of AVT on vocal behavior in this species are therefore not the same for each call type. AVT may act at more general motivational levels in the central nervous system and other neural or endocrine factors may control choice of call type and direct motor output.     

A preliminary study, using multivariate analysis, of a species complex of African house snakes (Boaedon fuliginosus)

systematics of Black and Lined house snakes from the Boaedon fuliginosus species complex were investigated using the limited amount of readily available material. These two forms have recently been regarded both as separate species, and as conspecific morphs. Multivariate and univariate biometrics indicate that in West Africa these two forms are separate species, B. fuliginosus (Black house snake) and B. lineatus (Lined house snake). However, in the rest of Africa these two forms appear to be the conspecific morphs of a single species which is provisionally and tentatively regarded as B. fuliginosus even though it is predominantly striped. There is evidence of "character shift" and this species complex may pose some interesting problems for the concepts of "gene pool cohesion" and "phenetic species".     

Male territoriality,fighting and assessment of fighting ability in the Australian frog Uperoleia rugosa

Males of Uperoleia rugosa form aggregations around the breeding pond. Within the aggregations, each calling male maintains an exclusive zone in which it does not tolerate another calling male. However, attacks on silent intruders, satellite males or females were never observed. Territorial males warned calling intruders by changing from advertisement calls to encounter calls (a display); if the intruder persisted in calling the resident would either attack or retreat and call elsewhere. Fights were usually won by the heavier males, and males of similar weight were more likely to fight, whereas males retreated from opponents that were substantially heavier. Calling was energetically costly and territorial males lost weight and thus their fighting abilities declined, whereas silent satellite males gained weight and were able to oust lighter territorial males. There was thus a dynamic interchange between territorial and satellite males depending on their relative weights. The playback of recorded advertisement calls showed that males can assess each others' fighting abilities on the basis of their vocalizations. Males retreated from a loudspeaker broadcasting the calls of a larger male but they attacked the loudspeaker if the calls of smaller or similar sized males were played (N=39, P<0·001, one-tailed binomial test). Dominant frequency was probably the call parameter used in assessment as it was correlated with the weight (r=−0·71, P<0·001, N=100) and hence the fighting ability of the caller, but this hypothesis needs further testing.     

Multisensory signals trigger approach behaviour in the fire-bellied toad Bombina orientalis: sex differences and call specificity

Animal communication often involves multimodal signals, and interactions between sensory modalities can trigger unique responses in receivers. Response to social signals was investigated in fire-bellied toads by exposing them to playback of male calls (advertisement and release calls) and a video clip of a male conspecific in the laboratory. The cues were presented in isolation and as a combined bimodal stimulus, and approach frequency, latency to approach and time spent around the stimulus source were measured. No positive phonotaxis was observed toward the advertisement call, both during the day and during a phonotaxis trial performed at night. However, females, but not males, approached with greater frequency, lower latency, and spent more time near the source of the bimodal stimulus in an experiment involving the advertisement call. Female response was specific to the advertisement call, as approach was not increased when the release call was used. Males, on the other hand, did not show increased approach in the advertisement call experiment, but approached with greater frequency the bimodal stimulus involving the release call within the first minute of stimulus presentation. The findings suggest that females orient toward calling males and that males eavesdrop on release calls, but in both cases a visual stimulus is also needed to trigger a response. Social approach in Bombina orientalis is thus dependent on multisensory cues, and the nature of the interaction between sensory modalities depends on receiver sex and call type.     

A New Species of the Genus Leptolalax(Anura: Megophryidae) from Northern Thailand

A new species of the genus Leptolalax is described from Doi Saket, Chiang Mai Province, northern Thailand. The new species is distinguished from its congeners by the following combination of characters: 1) adult size large(male SVL 45.8-52.5 mm); 2) tympanum distinct; 3) skin of dorsal surface mostly smooth, with distinct tubercles on dorsal and lateral parts of the body; 4) ventrolateral glandular ridge distinct and complete; 5) lateral body and groin covered with dark brown or black spots; 6) throat, chest, and belly nearly immaculate white; and 7) bi-colored iris, upper onethird golden, and lower two-thirds gray. Molecular phylogenetic analysis of mitochondrial 16S rRNA further distinguishes it from its congeners for which sequences are available.     

Male secondary sexual characteristics, sexual selection, and molecular divergence in fanged ranid frogs of Southeast Asia

A group of Southeast Asian frogs of the family Ranidae are characterized by a suite of unusual secondary sexual characteristics. Males are larger than females, possess bony odontoid processes (fangs) and have enlarged heads. While all members of this clade are sexually dimorphic in this suite of characters, the degree of development of the secondary sexual characteristics in males varies widely across species. One of the most common species, Rana blythii Boulenger, has an extremely broad geographical range. This frog lacks an advertisement call in Borneo but calling has been reported in populations from peninsular Malaysia and Vietnam. This study examines the relationship between the expression of male secondary sexual characteristics, molecular divergence and number of speciation events in widely separated populations of R. blythii , and between R. blythii and other species of closely related fanged frogs belonging to the grunniens species group. Results from morphometric and sequence data indicate that (1) there is no correlation between morphological and molecular distance in these frogs and (2) there is no correlation between morphological distance and the number of speciation events separating taxa. Systematic analysis demonstrates that R. blythii , as currently recognized, is not a single species nor is it monophyletic.     

Conserved karyotypes in the Hyla pulchella species group (Anura, Hylidae)

Cytogenetic analyses were done on specimens of Hyla marginata and on three populations of H. semiguttata differing in morphology and in the physical parameters of their advertisement call, as well as in individuals of Hyla sp. (aff. semiguttata). All specimens had 2n=24 chromosomes with a morphology very similar to that of other 24-chromosome Hyla species. Hyla semiguttata and H. marginata showed the same C-banding pattern but were distinguished by the location of the NOR on pair 1 in H. semiguttata (in the three populations) and Hyla sp. (aff. semiguttata), and on pair 10 in H. marginata. The H. semiguttata populations did not differ cytogenetically, despite variations in their morphology and advertisement calls. Similarly, H. semiguttata and H. p. joaquini studied previously had identical C-banding patterns and NOR locations, suggesting that they are very closely related.     

A developmental staging series for the African house snake, Boaedon (Lamprophis) fuliginosus

Embryonic staging series are important tools in the study of morphological evolution as they establish a common standard for future studies. In this study, we describe the in ovo embryological development of the African house snake (Boaedon fuliginosus), a non-venomous, egg-laying species within the superfamily Elapoidea. We develop our staging series based on external morphology of the embryo including the head, eye, facial prominences, pharyngeal slits, heart, scales, and endolymphatic ducts. An analysis of embryonic growth in length and mass is presented, as well as preliminary data on craniofacial skeletal development. Our results indicate that B. fuliginosus embryos are well into organogenesis but lack well-defined facial prominences at the time of oviposition. Mandibular and maxillary processes extend rostrally within 8 days (stage 3), corresponding to the first appearance of Meckel's cartilages. Overall, the development of the craniofacial skeleton in B. fuliginosus appears similar to that of other snake species with intramembraneous bones (e.g., dentary and compound bones) ossifying before most of the endochondral bones, the first of which to ossify are the quadrate and the otic capsule. Our staging series is the first to describe the post-ovipositional development of a non-venomous elapoid based on external morphology. This species is an extremely tractable captive that can produce large clutches of eggs every 45 days throughout the year. As such, B. fuliginosus should be a good model for evolutionary developmental biologists focusing on the craniofacial skeleton, loss of limbs, generational teeth, and venom delivery systems.     

A new golden frog species of the genus Diasporus (Amphibia, Eleutherodactylidae) from the Cordillera Central, western Panama

We describe the frog species Diasporus citrinobapheussp. n. from the Cordillera Central of western Panama. The new species differs from all other species in its genus in coloration, disk cover and disk pad shape, skin texture, advertisement call, and size. It is most similar to Diasporus tigrillo, from which it differs in dorsal skin texture, relative tibia length, number of vomerine teeth, ventral coloration, dorsal markings, and relative tympanum size, and to Diasporus gularis, from which it can be distinguished by the lack of membranes between the toes, adult size, posterior thigh coloration, and position of the choanae. We provide data on morpho- logy, vocalization, and distribution of the new species, as well as brief information on its natural history.     

Liparis rubescens sp. nov. (Orchidaceae) from Thailand

In the context of an ongoing revision of the orchid genus Liparis in Thailand, we describe and illustrate a new species under the name L. rubescens Tetsana, H. A. Pedersen & Sridith. Morphologically, it is particularly similar to L. dumaguetensis Ames from the Philippines, but it differs in the shape of the lateral sepals and in several characters of the labellum. Among the Liparis species already known from Thailand, L. atrosanguinea Ridl. bears a certain resemblance to the new species, but not so much as to make misidentifications likely. So far, L. rubescens is only known from Khao Nan National Park in the southern Thai province of Nakhon Si Thammarat, but we suggest that it should also be searched for in other parts of peninsular Thailand, southernmost Myanmar and Peninsular Malaysia.