Mate Choice by Chemical Cues in Western Redback (Plethodon vehiculum) and Dunn's (P. dunni) Salamanders

Western redback and Dunn's salamanders (Plethodon vehiculum and Plethodon dunni, respectively) can distinguish between potential mates by using chemical cues. In laboratory choice tests, adult males of both species showed significant discrimination between chemical cues of gravid females over non-gravid females of equal body size. Furthermore, males of both species differentiated the odour of paired gravid females that differed by ? 5 mm snout-vent length (SVL). Given that clutch size is related to female body size in these species, adult males may be able to distinguish between females via cues that signal potentially high female reproductive success. In choice tests, P. vehiculum females did not discriminate between two relatively large males that differed by ? 5 mm SVL. However, females of P. vehiculum did discriminate between two relatively small males that differed by the same amount. Apparently, P. vehiculum females ranked males by both absolute and relative body size using chemical cues. This pattern could reflect a female preference for large males or that females avoid mating with the smallest males.     

Underwater Video Reveals Strong Avoidance of Chemical Alarm Cues by Prey Fishes

A diversity of fishes release chemical cues upon being attacked by a predator. These cues, commonly termed alarm cues, act as sources of public information warning conspecifics of predation risk. Species which are members of the same prey guild (i.e. syntopic and share predators) often respond to one another's alarm cues. The purpose of this study was to discriminate avoidance responses of fishes to conspecific alarm cues and cues of other prey guild members from responses to unknown damaged fish odours and novel odours. We used underwater video to measure avoidance responses of freshwater littoral species, namely fathead minnows (Pimephales promelas), finescale dace (Chrosomus neogaeus), and brook stickleback (Culaea inconstans), to both injured fish cues and novel non‐fish odours. The cyprinids (minnows and dace) showed significant avoidance of minnow cues over swordtail cues, morpholine, and the control of distilled water and tended to avoid fathead cues over cues of known prey guild members (stickleback). Cyprinids also significantly avoided cues of stickleback over unknown heterospecific cues (swordtail) and tended to avoid stickleback cues over morpholine and the distilled water control. Stickleback significantly avoided fathead minnow extract over the distilled water and tended to avoid stickleback and swordtail over distilled water. We conclude that fishes in their natural environment can show dramatic changes in behaviour upon exposure to alarm cues from conspecifics and prey guild members. These responses do not represent avoidance of cues of any injured fish or any novel odour.     

The Use of Conspecific Chemical Cues for Cannibal Avoidance in California Newts (Taricha torosa)

Adult California newts, Taricha torosa (Caudata: Salamandridae), prey on both conspecific egg masses and larvae. We used both laboratory and field experiments to determine if newt larvae use conspecific chemical cues to avoid adult intraspecific predators. Larvae showed significant hiding responses when exposed to adult cues in both the field and the laboratory. Larvae did not exhibit significant hiding responses to chemical cues from invertebrate predators.     

Population and Species Divergence of Chemical Cues that Influence Male Recognition of Females in Desmognathine Salamanders

Growing evidence indicates that males may be more discriminating of mating partners than often has been assumed. In the North American Ocoee dusky salamander, Desmognathus ocoee (Plethodontidae: Desmognathinae), sexual incompatibility among conspecific populations is high in encounters staged in the laboratory, at least in part because males fail to recognize ‘other’ females as appropriate targets for courtship. I used Y-mazes to test the hypothesis that males of D. ocoee discriminate between substrate-borne chemical cues produced by ‘own’ (homotypic) and ‘other’ (heterotypic) females. Males of four populations discriminated in favor of substrates soiled by homotypic females over clean (control) substrates (expt 1), suggesting that females produce chemical cues of sociosexual significance to males. Furthermore, males from these populations discriminated in favor of substrates soiled by homotypic females vs. substrates soiled by heterotypic females (expt 2), both conspecific and heterospecific (D. carolinensis and D. orestes). Thus, differences among populations and species in female chemical cues appear to affect the chemotactic responses of males. I suggest that, together with differences in behavioral signals and responses exhibited during courtship, differences in female chemical cues likely contribute to sexual incompatibility among populations and taxa of desmognathine salamanders.     

Chemical Alarm Cues Are Conserved within the Coral Reef Fish Family Pomacentridae

Fishes are known to use chemical alarm cues from both conspecifics and heterospecifics to assess local predation risks and enhance predator detection. Yet it is unknown how recognition of heterospecific cues arises for coral reef fishes. Here, we test if naïve juvenile fish have an innate recognition of heterospecific alarm cues. We also examine if there is a relationship between the intensity of the antipredator response to these cues and the degree to which species are related to each other. Naïve juvenile anemone fish, Amphiprion percula, were tested to see if they displayed antipredator responses to chemical alarm cues from four closely related heterospecific species (family Pomacentridae), a distantly related sympatric species (Asterropteryx semipunctatus) and a saltwater (control). Juveniles displayed significant reductions in foraging rate when exposed to all four confamilial heterospecific species but they did not respond to the distantly related sympatric species or the saltwater control. There was also a strong relationship between the intensity of the antipredator response and the extent to which species were related, with responses weakening as species became more distantly related. These findings demonstrate that chemical alarm cues are conserved within the pomacentrid family, providing juveniles with an innate recognition of heterospecific alarm cues as predicted by the phylogenetic relatedness hypothesis.     

Social Context Influences the Antipredator Behaviour of Fathead Minnows to Chemical Alarm Cues

Prey fishes, like many organisms under fluctuating predation threat, rely on multiple sources of information to accurately gauge current risk. This includes the use of chemical cues such as alarm cues from damaged conspecifics or familiar heterospecifics, as well as the odour of known predators. While each fish is well equipped with its own array of sensory abilities, they should also be alert to the behaviours of nearby neighbours who may have information they lack. In the present study, we tested the ability of fathead minnows to use social cues in combination with the odour of damaged conspecifics and heterospecifics to mediate the assessment of predation risk. Specifically, we tested whether the presence of a shoal of conspecifics or familiar heterospecifics would significantly change a minnow's antipredator behaviour when exposed to the odour of a damage‐release cue from a conspecific or ecologically similar heterospecific. The results of our study showed a significant interaction between the damage‐release cues to which the minnows were exposed and the presence/absence of shoalmates. These findings have important implications for the design of future investigations of antipredator responses because most studies of group‐living prey have been conducted on solitary subjects.     

Fish Faeces as the Primary Source of Chemical Cues Inducing Fish Avoidance Diapause in Daphnia Magna

In a laboratory batch culture experiment, females of Daphnia magna were exposed to five different experimental media containing either:(1) water from an aquarium with fish, (2) extract of fish faeces, (3) mixture of both media, (4) extract of homogenised conspecific Daphnia, or (5) control water without the addition of extra cues. The experiment was planned to test potential pathways of excretion of the chemical cues that induce resting-egg formation in D. magna and to find an effective way of collecting these chemical cues. The results indicate that fish faeces are the prevailing source of the chemical cues that induce resting-egg production in D. magna. The ease of collection and the possibility of storing it in a frozen state make it a convenient cue for inducing diapause response in Daphnia. The results of the experiment imply that in natural conditions Daphnia may face high concentration of the inductive signals once migrating to the bottom zone where fish faeces commonly accumulate.     

Thermal Sensitivity of Aggression in Two Terrestrial Salamanders,Plethodon cylindraceus and P. montanus

Understanding the abiotic and biotic mechanisms regulating species distributions is an important area of study for ecologists. Increasingly, studies have begun to focus on the influence of abiotic conditions on biotic interactions. Numerous examples of interspecific aggression exist within the terrestrial salamander genus Plethodon and many occur between montane endemics and lowland conspecifics. Here, we explore how variation in temperature might influence aggressive interactions between two Plethodon species that have relatively broad elevational overlap, P. cylindraceus and P. montanus. We found that both species increased aggressive acts with increasing temperatures. We also found that the pattern of variation in aggression across test temperatures differed between species. Interestingly, we found that the lowland congener, P. cylindraceus, was more aggressive than the montane endemic, P. montanus, a pattern opposite of previous observations within Plethodon. The large range of sympatry observed between these species may be a result of a reverse of what is expected based on previous research with congeners in similar systems and not the result of temperature‐specific outcomes.     

The Behavioural Response of Mayfly Nymphs (Stenonema sp.) to Chemical Cues from Crayfish (Orconectes rusticus)

Artificial reef systems play an important role in the increase of natural production of biological marine resources and they have been deployed worldwide. In Portugal, seven artificial systems have been deployed along the southern coast of the Algarve. Research to date has focussed mainly on fish populations, particularly those of economical importance. The present work aims to study the macrobenthic communities of the artificial reef structures, as these will enhance the food resources and shelter, making the reefs more attractive to fish. In particular, we experimentally analysed the sequence of colonisation of macrobenthic communities of the Ancão artificial reef system, which was deployed in the summer of 2002. The study of the colonisation of benthic communities was done using samples of concrete cubic units (15 × 15 cm) that were suspended at the reef modules at a depth of 20 m, at the time of the reef immersion. Four replicate samples were collected by SCUBA diving from two groups of the Ancão reef every three months from the starting date. Sampling was done using essentially non-destructive methods to assess the percentage cover of macrobenthic organisms in both vertical and horizontal surfaces. The percentage cover of the taxonomic groups was compared within the different surfaces of the samples and between the two reef groups. The bottom surface of cubic samples had a significantly higher colonisation related to the dominance of barnacle cover, probably due to lower sedimentation levels. Samples from both reef groups showed a similar pattern of colonisation. Barnacles, bryozoans and serpulids dominated the samples three months immediately after the beginning of the experiment. Other invertebrates groups, such as Porifera, Hydrozoa, Anthozoa, other sessile Polychaeta, Decapoda, Gastropoda and Bivalvia, were more abundant after six months of colonisation.     

Fathead Minnows, Pimephales promelas, Learn to Recognize Chemical Alarm Cues of Introduced Brook Stickleback, Culaea inconstans

In four experiments conducted over a 6-year period, we investigated whether fathead minnows, Pimephales promelas, could acquire the ability to recognize chemical alarm cues of introduced brook stickleback, Culaea inconstans. A laboratory experiment documented that stickleback-naïve minnows did not exhibit an anti-predator response when exposed to the chemical alarm cues of stickleback. In a laboratory experiment conducted 5 years after the introduction of stickleback to the pond, minnows exhibited an antipredator response to stickleback cues. Moreover, in a field experiment the minnows exhibited avoidance of areas labelled with stickleback alarm cues. Minnows raised from eggs taken from the test pond did not exhibit an anti-predator response to stickleback cues while minnows from the test pond that had experience with stickleback cues did respond to stickleback cues. Our results provide clear evidence that cross-species responses to chemical alarm cues of fishes can be learned. Learned recognition of alarm cues has important implications for predator/prey interactions.     

Sex Recognition via Chemical Cues in the Sex-Role-Reversed Gulf Pipefish (Syngnathus scovelli)

Sexual selection theory predicts that members of the choosy sex, usually females, should employ multiple sensory systems to obtain information about potential mates. Such predictions should also apply to systems in which sexual selection acts most strongly on females (i.e. sex-role-reversed species), and males of these taxa should be the individuals employing multiple cues to assess female attractiveness. Very little work has been directed toward mate choice involving multiple sensory systems in sex-role-reversed taxa, but fishes of the family Syngnathidae (pipefishes, seahorses, and sea dragons) provide an excellent opportunity to contribute to this research enterprise. While much is known about visual communication in pipefish, the role of chemical communication has not been investigated. Using dichotomous choice tests, we found that male, but not female, Gulf pipefish attended to chemical cues of opposite sex conspecifics. Given that males distinguished sex on the basis of chemical cues, we also tested whether males could assess female body size, an important trait with respect to mate choice in pipefish, on the basis of chemical cues alone. When given the choice between chemical cues produced by large vs. small females, males exhibited no preference. Our results suggest that male pipefish can use chemical cues to distinguish between males and females but not to differentiate females of different body size.     

The nucleolar organizer of Plethodon cinereus cinereus (Green)

The lampbrush nucleolar organizers in P. c. cinereus are on the shorter arms of the 7th longest bivalent, near to the centromeres. The organizer consists of a non-chromomeric, Feulgen-negative section of the half bivalent axis inserted between typically chromomeric regions with normal lampbrush loops. The main axis of the organizer may appear double or single. The distributions and lengths of the double regions are variable. The axial strand of the organizer can be broken with DNase. Nucleolus-like objects are attached at irregular intervals along the organizer axis. The attached nucleoli closely resemble the free nucleoli in the same nucleus. Where free and attached nucleoli appear as beaded rings, the general characteristics and range of lengths of the attached rings are similar to those of the free rings. The attached rings occur singly, in pairs, or in clusters. The point of attachment of a ring is sometimes marked by a granule on the organizer axis. Pairs of attached nucleolar rings sometimes form double bridges in which the nucleoli extend linearly across a gap in the organizer axis. The length of the organizer varies from 20 μm to 300 μm, depending, at least in part, on the stage of oocyte development. The difference is a function of the length of the axial strand. Nucleolar organizers similar to that described for P. c. cinereus have been seen on the lampbrush bivalents of 7 other species of Plethodon and 2 species of Eurycea. In all of these, the 2 organizers of the nucleolus bivalents differ in length.     

The nucleolar organizer of Plethodon cinereus cinereus (Green)

The 7th longest lampbrush bivalent in oocytes of Plethodon cinereus cinereus has a region of attached oocyte nucleoli near to the centromere on the shorter arm of each half bivalent. When squash preparations of P. c. cinereus spermatocytes are treated with NaOH to denature chromosomal DNA, and subsequently incubated in a solution of (³H) ribosomal RNA from Xenopus cell cultures, the ribosomal RNA binds specifically to a region near the centromeres on the shorter arms of the 7th longest bivalent, and to a region near the end of the shorter arm of the 14th bivalent. The amount of ribosomal RNA bound to the 7th bivalent at diplotene and 1st meiotic metaphase is regularly different on the 2 halves of the bivalent. Each half of the 14th bivalent is usually labelled more heavily than the less heavily labelled half of the 7th bivalent. These observations are discussed in relation to the involvement of nucleolar organizers in gene amplification and rectification.     

Isolation of dinucleotide microsatellite loci from red‐backed salamander (Plethodon cinereus)

We isolated 13 variable dinucleotide microsatellites from red‐backed salamanders (Plethodon cinereus). After generating fragments using degenerate oligonucleotide primer‐polymerase chain reaction (DOP‐PCR), AC repeats were captured using biotinylated probes and streptavidin‐coated magnetic particles. Captured fragments were cloned into plasmids, screened for microsatellites with a simple PCR reaction, and select plasmids then sequenced. PCR primers were designed and optimized for robust amplification, and nine primers have been further optimized for multiplex reactions with fluorescent primers. These nine loci are variable with an average of 6.11 alleles per locus and an average heterozygosity of 0.61.     

Female Annual Killifish Austrolebias reicherti (Cyprinodontiformes,Rivulidae) Attend to Male Chemical Cues

Chemical communication is an important mediator of social interactions in fish, including mate recognition, mate choice and intrasexual competition. We used a simultaneous‐choice design to test the response of adult females of the annual killifish, Austrolebias reicherti, towards male‐conditioned water, both crude and fractionated into its polar and non‐polar components. Females preferred male‐conditioned water when tested both against control water and female‐conditioned water. The polar and non‐polar fractions of the male‐conditioned water were not significantly attractive, although a tendency was observed towards the non‐polar fraction. Our study shows that annual fish respond to the chemical cues of potential mates. The chemical channel is likely to be particularly important in the muddy ephemeral ponds in which they occur.     

北方山溪鲵精巢显微结构的年周期变化

2001、2003年1～12月自秦岭北坡的溪流中,共采集北方山溪鲵(Batrachuperus tibetanus)38尾(体重27～38g)做精巢切片,在光镜下观察精巢显微结构的年周期变化。结果表明：北方山溪鲵为非连续型精子发生,精原细胞增生有两个高峰期,分别在9～11月和翌年的3～5月,初级精母细胞的成熟分裂期在6月末至7月,精子形成期在7月末至8月。每个精巢小叶可明显地分为深层的非成熟区和浅层的成熟区。非成熟区为精原细胞的贮存区,可增殖并延伸为增殖区。增殖区再发育为成熟区,为精子形成和存储的区域。在精子排出后,成熟区转变为排空区,后者被新的增殖区更替。北方山溪鲵精巢所有小叶同步发育,无精子成熟波。     

Systematic Relationships of Hynobius okiensis among Japanese Salamanders (Amphibia: Caudata)

We conducted an electrophoretic survey to examine systematic relationships of a lotic-breeding salamander Hynobius okiensis endemic to Dogo Island of the Oki Islands, Japan, with several lentic and lotic-breeding Japanese species. Genetically H. okiensis with 2n=56 chromosomes was closer to the lentic-breeding H. nebulosus group (H. nebulosus and H. dunni) with the same chromosome number than to the lotic-breeding H. naevius group (H. naevius and H. kimurae) and H. boulengeri with 58 chromosomes. Chromosome number reduction from 58 to 56, possibly accompanied with a change in breeding environment from streams to still waters, is estimated to have first occurred in the nebulosus group of Hynobius. A reversal only in breeding habits then seems to have followed in steep, montane environments of the small island of Dogo, resulting in the speciation of H. okiensis.