Reproductive interference between Rana dalmatina and Rana temporaria affects reproductive success in natural populations

Experimental evidence suggests that reproductive interference between heterospecifics can seriously affect individual fitness; support from field studies for such an effect has, however, remained scarce. We studied reproductive interference in 25 natural breeding ponds in an area where two ranid frogs, Rana dalmatina and Rana temporaria, co-occur. The breeding seasons of the two species usually overlap and males of both species are often found in amplexus with heterospecific females, even though matings between heterospecifics produce no viable offspring. We estimated species abundance ratios based on the number of clutches laid and evaluated fertilization success. In ponds with low spatial complexity and a species abundance ratio biased towards R. temporaria, the average fertilization success of R. dalmatina eggs decreased, while this relationship was not detectable in spatially more complex ponds. Fertilization success of R. temporaria did not decrease with increasing relative numbers of heterospecifics. This asymmetry in fitness effects of reproductive interference may be attributed to R. temporaria males being more competitive in scramble competition for females than R. dalmatina males. Our study is among the first to demonstrate that in natural breeding populations of vertebrates interference among heterospecifics has the potential to substantially lower reproductive success at the population level, which may in turn affect population dynamics.     

Lymphoma development in mice chronically exposed to UMTS-modulated radiofrequency electromagnetic fields

There are public concerns regarding possible carcinogenic or cancer-promoting effects of electromagnetic fields (EMFs) from mobile phones and base stations. The objective of the present study was to investigate whether chronic exposure to EMFs of the UMTS (Universal Mobile Telecommunication System) influences the development of lymphoma in a lymphoma animal model, the AKR/J mouse. Unrestrained mice were chronically sham-exposed (n = 160) or exposed (n = 160) in identical exposure systems (radial waveguides) to a generic UMTS test signal (24 h per day, 7 days per week, 0.4 W/kg SAR). Additionally, 30 animals were kept as cage controls. Animals were checked visually each day and were weighed and palpated weekly to detect swollen lymph nodes. Starting at the age of 6 months, blood samples were taken from the tail every 2 weeks to perform differential leukocyte counts and to measure the hematocrit. Visibly diseased animals or those older than 43 weeks were killed humanely, and tissue slices were examined for metastatic infiltrations and lymphoma type. The study was performed in a blinded way. Cage control animals had a significantly lower growth rate than those kept in the radial waveguides. The number of ill animals, the mean survival time, and the severity code of the disease did not differ between the experimental groups. Therefore, the data show no negative effects from exposure and corroborate earlier findings in AKR/J mice exposed to GSM EMF (Sommer et al., BMC Cancer 4, 77-90, 2004).     

Population structure and genetic diversity of Rana dalmatina in the Iberian Peninsula

The increasing fragmentation of natural habitats may strongly affect patterns of dispersal and gene flow among populations, and thus alter evolutionary dynamics. We examined genetic variation at twelve microsatellite loci in the Agile frog (Rana dalmatina) from 22 breeding ponds in the Iberian Peninsula, the southwest limit of its range, where populations of this species are severely fragmented and are of conservation concern. We investigated genetic diversity, structure and gene flow within and among populations. Diversity as observed heterozygosities ranged from 0.257 to 0.586. The mean number of alleles was 3.6. Just one population showed a significant F _IS value. Four populations show evidence of recent bottlenecks. Strong pattern of structure was observed due to isolation by distance and to landscape structure. The average degree of genetic differentiation among populations was F _ST = 0.185. Three operational conservation units with metapopulation structure were identified. Additionally, there are some other isolated populations. The results reinforce the view that amphibian populations are highly structured even in small geographic areas. The knowledge of genetic structure pattern and gene flow is fundamental information for developing programmes for the preservation of R. dalmatina at the limits of its geographic distribution.     

Embryonic and larval defensive responses of agile frog (Rana dalmatina) to alien crayfish

Red swamp crayfish Procambarus clarkii, a widespread invasive alien crayfish, represents a serious threat for several freshwater species, including amphibians, which are declining at a global scale. As a shared coevolutionary history is the main factor determining the emergence of antipredator responses, Anuran tadpoles may not be able to cope effectively with this introduced predator. We performed two experiments to assess agile frog's (Rana dalmatina) defensive responses to both P. clarkii and native dragonfly larvae (Anax imperator). First, we conditioned embryos (collected from two ponds 30 km away from each other) with predators’ chemical cues to explore possible variation in hatching time caused by predation risk. In the second experiment, to evaluate how predators’ diet affects tadpole behavior, we conditioned tadpoles for a 5‐week period with cues from tadpole‐fed and gammarid‐fed predators and recorded behavioral and morphological responses. Embryos did not alter hatching time in the presence of any predator cue, while tadpoles from both populations strongly reduced activity and visibility when raised in the presence of tadpole‐fed dragonfly larvae. Morphological changes were less straightforward and were induced only in one population, for which broader tails and a slight increase in body size of tadpoles exposed to tadpole‐fed predators were observed. The lack of defensive responses in crayfish‐exposed tadpoles suggests that the spreading of this invasive species in agricultural lowlands of northern Italy may represent a further threat to their conservation.     

Role of aromatase and androgen receptor expression in gonadal sex differentiation of ZW/ZZ-type frogs,Rana rugosa

To elucidate the mechanisms of amphibian gonadal sex differentiation, we examined the expression of aromatase and androgen receptor (AR) mRNAs for days 17-31 after fertilization. The effects of inhibitors and sex steroid hormones were also examined. In ZZ males, expression of AR decreased after day 19, while aromatase expression was low throughout the sampling period. Males treated with 17beta-estradiol (E2) showed increasing aromatase expression after day 21, and formed ovaries. AR antagonist treatment also induced high-level aromatase expression and ovarian differentiation. In males co-treated with an aromatase inhibitor and E2, the undifferentiated gonads developed into testes despite high-level aromatase expression. Males treated with androgen and E2 before and during an estrogen sensitive period, respectively, also formed testes. In ZW females, AR expression persisted at a low-level, while aromatase expression increased after day 18. Short-term treatment with an aromatase inhibitor was ineffective in preventing ovarian differentiation, whereas long-term treatment resulted in testes developing from ovarian structure. Compared with the ZZ males and ZW females, WW females did not exhibit detectable expression of AR, suggesting that the active AR gene(s) itself, or a putative gene regulating AR gene expression, is located on Z chromosomes. From the time lag of aromatase expression between ZW females and ZZ males treated with E2 and the effect of AR antagonist, it was found that in males elevated AR expression suppresses aromatase expression directly or indirectly. Consequently, endogenous androgens, accumulated by blocking estrogen biosynthesis, induced testicular differentiation. The gonadogenesis of males is dependent on sex hormone, whereas that of females has evolved to hormone-independence.     

Pond and landscape determinants of Rana dalmatina population sizes in a Romanian rural landscape

Amphibians are good indicators of human impact, declining steadily worldwide. We explored the relationships between the ponds and nearby landscape parameters and population size of the Agile Frog (Rana dalmatina), estimated from the number of egg masses, in a cultural landscape within the central section of Târnava Mare Basin, Romania. Forty-three permanent ponds were surveyed in a 2600 km² area. The average number of egg masses per pond was 211.13 (SD = 426.41). The egg mass number was significantly and positively related to the emergent aquatic macrophyte cover (its effect peaks at around 50%) and the green connecting corridors between the ponds and forests, and negatively related to the extent of nearby urban areas. The proximity of the forest (positive effect) and the presence of high traffic roads (negative effect) were highly correlated with green corridors and further eliminated from the model due to multicollinearity. Both these variables had significant effects when incorporated in univariate models and multivariate models without green corridors. Since a large part of our study area was currently declared as Natura 2000 site, there is an increased need for management proposals and conservation applications for biodiversity, including amphibians. Rana dalmatina is an important species for monitoring because it is common in the studied area and is suited for short surveys.     

Inbreeding and road effect zone in a Ranidae: the case of Agile frog, Rana dalmatina Bonaparte, 1840

Inbreeding has often been invoked in the extinction of local populations. In eleven western France populations of Agile frog studied, observed heterozygosity was significantly lower than expected in all cases, giving new evidence of such a depression in small populations. It especially occurred in ponds located near an highway rather than in undisturbed populations (FIS = 0.544 and 0.315, respectively). Thus, our results argue for a "road effect zone". Discussing about road distance and conservation policies, we propose that roads are directly involved in inbreeding and in local extinction. Thus, road construction ought to consider conservation management.     

Asynchronous arrival pattern, operational sex ratio and occurrence of multiple paternities in a territorial breeding anuran, Rana dalmatina

Understanding why females mate multiply is a major issue in evolutionary ecology. We investigated the consequences of an asynchronous arrival pattern on male competition and multiple paternity in the apparently monoandrous agile frog ( Rana dalmatina ). The largest frogs arrived first and both males and females lost weight significantly during the spawning period. Asynchronous arrival at breeding sites resulted in a male-biased operational sex ratio (OSR). The OSR was more strongly male-biased at the beginning and at the end of the breeding period when the number of satellite males increased. All females mated only once, but multiple paternity within clutches occurred at the beginning and the end of the breeding period. The influence of asynchronous arrival and biased sex ratio suggests that reduced variance or bet-hedging promoting female fitness had only a reduced role in the evolution of polyandry, and polyandry is likely to be associated with male benefits. Polyandry in frogs can be explained either by forced mating as a result of sexual conflict or by clutch piracy. By modifying intrasexual competition, asynchronous arrival and changes in OSR may have a decisive influence upon the evolution of mating systems and favour both polyandry and stable coexistence of alternative mating behaviour.  © 2005 The Linnean Society of London, Biological Journal of the Linnean Society , 2005, 86 , 191–200.     

Blood pressure in monkeys chronically exposed to psycho-emotional stress

The influence of experimental neurosis due to repeated conflict situations on blood pressure was studied in male monkeys. All animals developed hypertension demonstrable in the conscious state but which disappeared under pentobarbital anaesthesia. Both in conscious and anaesthetized animals, the pulse pressure was in-elevated. The hypertension was accompanied by increased plasma volume to interstitial volume ratio, due to a decrease of the interstitial fluid compartment. The plasma renin concentration was not raised. It is suggested that chronic increase in blood pressure might be responsible for the decrease in compliance of large arteries (as evidenced by increased pulse pressure). This form of hypertension does not depend on the activity of the renin-angiotensin system.     

Reduction in open field-induced hyperthermia in the rat exposed to chlorpyrifos,an anticholinesterase pesticide

Rodents develop a marked elevation in core temperature (T_c) when they are placed in an open field environment (OFE). Open field-induced hyperthermia (OFH) involves activation of thermoregulatory systems that are quiescent under baseline conditions. Cholinergic stimulation arising from exposure to organophosphates (OP) is expected to modulate OFH; subtle effects of OPs may be observed during OFH that would not occur in the absence of stress. We have found that methyl scopolamine (MS), a peripheral muscarinic antagonist, attenuates stress-induced hyperthermia. To this end, we assessed if a low dose of the OP pesticide chlorpyrifos (CHP) would alter OFH with and without administration of MS. T_c and motor activity (MA) of male, Sprague-Dawley rats were monitored by telemetry while housed in their home cage at an ambient temperature of 22°C. At 9 AM the rats were gavaged with corn oil or 10 mg/kg CHP that resulted in a ∼40% inhibition in plasma cholinesterase activity. T_c and MA were the same in the control and CHP groups prior to OFE. The rats were then dosed IP with saline or 1.0 mg/kg MS at 12 noon and subjected to OFE (61×61×61 cm box) for 1 h while T_c and MA were monitored every 2 min. T_c of the corn oil/saline group increased by 1.2°C during OFE, whereas T_c of the CHP/saline group was significantly attenuated. Administration of MS attenuated the OFH in rats treated with corn oil and CHP. Overall, exposure to OFE exacerbated the effects of a relatively low dose of CHP that had no effect in the unstressed animal. In addition, the OFH response to MS suggests a peripheral (i.e., outside of CNS) cholinergic pathway is operative in the control of OFH.