ALARM CALLS OF THE EUROPEAN GROUND SQUIRREL SPERMOPHILUS CITELLUS AND THE TAURUS GROUND SQUIRREL S. TAURENSIS ENCODE INFORMATION ABOUT CALLER IDENTITY

ABSTRACT

Vocalizations of many mammalian species have been reported to encode information about caller identity. In this study, we analyzed 300 alarm calls from 10 free-living European Ground Squirrels Spermophilus citellus (30 per individual) and 300 alarm calls from 10 free-living Taurus Ground Squirrels S. taurensis (30 per individual), and tested the potential of these calls to encode information about the callers' identities. Discriminant analysis including all 10 European Ground Squirrel individuals correctly classified 98% of calls, and cross-validation reached a classification success of 97%. Correct classification of 98% and cross-validation of 98% was assigned when the analysis included only those individuals producing calls consisting of both elements (eight individuals). For the Taurus Ground Squirrel, correct classification was 95% and cross-validation 94% for all 10 animals. When only those individuals producing calls consisting of both elements were included (eight individuals), discriminant analysis led to 94% correct classification and cross-validation produced a classification success of 93%. These analyses demonstrate that the structure of alarm calls in these two closely related species is highly variable and that it has significant potential to encode information about caller identity.     

Species-specific and shared features in vocal repertoires of three Eurasian ground squirrels (genus <Emphasis Type="Italic">Spermophilus</Emphasis>)

Along to alarm calls, Eurasian ground squirrels of the genus Spermophilus also produce other call types toward potential predators and rival conspecifics. Individually identified 50 speckled (Spermophilus suslicus), 18 European (S. citellus) and 59 yellow (S. fulvus) ground squirrels were examined for interspecies differences in their vocal repertoires. A separate sample of 116 (90 adult and 26 juvenile) S. suslicus was examined for presence of ultrasound in their alarm calls. In addition, all tonal calls in all the three species were checked for presence of nonlinear phenomena. Calls were elicited by approaching animals in live-traps or near burrows; some types of vocalizations were also recorded during handling. Eight call types, three tonal and five wideband ones, were described. Vocal repertoires were remarkably similar between species, excluding the alarm calls, which were species-specific. Alarm calls with ultrasonic components were found in two individuals of S. suslicus. Concerning nonlinear phenomena, biphonation in alarm calls of S. suslicus, frequency jumps and sidebands in screams of S. citellus, frequency jumps and subharmonics in screams of S. fulvus were found. Results are discussed with literature evidence on audible and ultrasonic vocalizations in ground squirrels.     

CONFERENCE ANNOUNCEMENTS

ABSTRACT

In most mammals, adults produce relatively low frequency vocalizations compared to those of juveniles. This rule is not maintained however at least in four species of ground squirrels, whose juveniles call at the adult's fundamental frequency. These findings have been obtained however with separate sets of juveniles and adults and no data is available concerning the ontogeny linked to these differences. Here we analyze the acoustic structure of alarm calls of the same Yellow Spermophilus fulvus and Speckled S. suslicus ground squirrel individuals, recorded as pups and as adults after hibernation. We found the fundamental frequencies of adults within the same frequency ranges as those of pups, in spite of the significant difference in body mass. In ground squirrels, severing the relationship between body size and call frequency removes some vocal cues to age. We discuss some functional hypotheses advanced to explain manipulations with fundamental frequencies in ground squirrels and other animals, and suggest the lack of data for discussing the mechanisms of such vocal tuning.     

Characteristics of arctic ground squirrel alarm calls

Summary Arctic ground squirrels, Citellus undulatus, produce six distinctly different sounds. Each of these sounds may represent a signal in itself, but combinations of these acoustic elements or repetition of a single element produce additional signals. Several of these signals serve as alarm calls. One sound element consists of a short (0.05 sec) broad frequency chat while another is a longer (0.16 to 0.25 sec) descending narrow frequency whistle. Squirrels utter three-note chatter calls when approached by a ground predator, and a series of five or more chatters, which fade in intensity, is given upon the close approach of a ground predator as the squirrel escapes into a burrow. A single whistle, which resembles the alarm call of some birds, is given upon the approach of an aerial predator. This call is repeated at approximately six to eight second intervals if the predator alights near a squirrel and remains nearby.     

Population genetic structure of the European ground squirrel in the Czech Republic

The European ground squirrel (Spermophilus citellus) is considered an endangered species with declining numbers throughout Europe, most pronounced at the western margin of its distribution area. Being extinct in Germany and Poland, the western margin of its distribution is in the Czech Republic. Here, landscape fragmentation has restricted the ground squirrels into few and very isolated localities where local extinctions still occur. In the present study we analysed European ground squirrels from six Czech and one Slovak localities using five microsatellite loci as genetic marker. The results show a strong genetic differentiation among the investigated populations (mean value of F _ST = 0.16) and high levels of inbreeding (values of F _IS ranged from 0.34 to 0.90). High level of inbreeding is generally considered to affect the viability of each population, which could lead to extinction. One of the most important factors is the lack of migration due to the large distances between the populations and the presence of migration barriers. Based on the results obtained we recommend a few suggestions for a conservation management of this species.     

Topographic distribution and daily activity of the European ground squirrel population in Bugacpuszta, Hungary

Factors affecting the population density and daily pattern of activity of the European ground squirrelSpermophilus citellus (Linnaeus, 1766) were studied in two habitats of the Kiskunság National Park, Hungary. One habitat was dominated by sandy and the other by salty grassland. Population densities were estimated by counting ground squirrel burrow entrances in sample areas with different microhabitat characteristics. Three categories (elevated, level and depressed) were used to describe the spatial distribution of burrows according to microtopography. Daily activity was evaluated by counting the ground squirrels visible on the surface and registering their alarm calls. The highest proportion of ground squirrel holes was found on elevations in the two habitats. Ground squirrels showed a similar daily activity pattern in different areas and in the same season in consecutive years. The daily activity consisted in a morning peak, a midday pause and a less regular afternoon activity period. Human activity had no direct effect on the daily activity pattern. Emission of alarm calls peaked in June, when litters emerge from the maternal burrows.     

Eavesdropping of an African ground squirrel on the heterospecific alarm calls of a noisy ground-nesting bird

Animals gather information about their environment from a variety of sources to enable adaptive decision-making behaviour. Eavesdropping on heterospecific alarm calls enhances predator avoidance, reduces time spent vigilant and allows for more time on daily activities such as foraging. If the information is relevant and reliable, individuals that respond to heterospecific signals may benefit from a wider range of information at a low marginal cost. The Cape ground squirrel (Xerus inauris) and crowned lapwing (Vanellus chilensis) are ground-dwelling species that are taxonomically distant but share similar predators, habitat and anti-predatory behaviours. We used playback experiments of the alarm calls produced by conspecifics and lapwings to investigate the vigilance responses of adult female Cape ground squirrels. Squirrels responded with greater vigilance to both squirrel and lapwing alarm calls, and no changes of vigilance levels were observed in response to a control sound. However, contrary to our predictions, changes in vigilance and time to relax did not differ between conspecific versus heterospecific playbacks. The results from our study suggest that squirrels perceive lapwing alarm calls as relevant and reliable information and that responding to it could increase their survival.     

冬眠对达乌尔黄鼠盲肠菌群的影响

冬眠哺乳动物的肠道微生物会发生季节性变化,同时在冬眠期间动物处于禁食状态,对肠道微生物的多样性和组成也产生影响。本研究通过16S rRNA基因高通量测序分析达乌尔黄鼠育肥阶段 (起始育肥期、快速育肥期、育肥完成期) 和冬眠阶段 (冬眠早期、冬眠晚期、出眠期) 共6个时期盲肠菌群的多样性、组成和功能,并通过冗余分析 (RDA) 探究其生理特征与菌群组成和功能之间的关系,揭示达乌尔黄鼠盲肠菌群的季节性变化。菌群组成的分析显示达乌尔黄鼠盲肠菌群主要由厚壁菌门 (Firmicutes)、拟杆菌门 (Bacteroidetes) 和疣微菌门 (Verrucomicrobia) 组成。与其他时期相比,冬眠早期厚壁菌门的相对丰度减少,拟杆菌门和疣微菌门的相对丰度增加。在Alpha多样性中,起始育肥期、快速育肥期和冬眠早期的Chao1和ACE指数显著低于出眠期,育肥完成期的Simpson指数显著低于快速育肥期 (P < 0.05) 。通过加权和非加权的UniFrac距离矩阵的主坐标分析发现盲肠菌群均显示出了明显的季节性聚类。PICRUSt分析中,丁酸代谢等代谢通路在育肥阶段富集,冬眠阶段集中在氮代谢等相关通路中。RDA分析显示达乌尔黄鼠不同时期的生理特征与其盲肠菌群的组成和功能显著相关。本研究表明,冬眠使达乌尔黄鼠盲肠菌群的多样性和相对丰度发生改变,盲肠菌群组成和功能的变化调节了达乌尔黄鼠的生理代谢,使达乌尔黄鼠适应季节性的环境变化。     

Effects of hibernation on somatostatin-like immunoreactivity in the brain of the ground squirrel (Spermophilus richardsonii) and European hedgehog (Erinaceus europaeus)

Summary The localization of the somatostatin system in the brains of Richardson's ground squirrels (Spermophilus richardsonii) and European hedgehogs (Erinaceus europaeus) was described by use of immunocytochemical methods. In addition, (i) chemically differing types of somatostatin and (ii) different activity phases of the somatostatin system during the hibernation cycle were investigated in the ground squirrel by means of high pressure liquid chromatography (HPLC) and radioimmunoassay (RIA).In both species, the hypothalamic component of the somatostatin system (periventricular nuclei, fiber projections to the median eminence) is more prominent than the widespread extrahypothalamic representation of the system displaying mainly scattered perikarya and nerve fibers. The reactivity pattern of the somatostatin system varied among hibernating, aroused, and non-hibernating animals; moreover, the interspecific differences were pronounced. The activity of the hypothalamic somatostatin system in the hibernating ground squirrel appeared to be suppressed when compared to non-hibernating controls, whereas in the hibernating hedgehog this system showed signs of increased activity in comparison to non-hibernating controls. In contrast, in the present material the extrahypothalamic components of the somatostatin system did not exhibit significant changes in their activity.     

Multigenic and morphometric differentiation of ground squirrels (Spermophilus, Scuiridae, Rodentia) in Turkey, with a description of a new species

This study reports on the molecular phylogenetics of ground squirrels, genus Spermophilus, in Turkey using cytochrome b (1140bp), part of the D-loop and flanking tRNAs (572bp), X chromosome (867-1051bp) and Y chromosome (983-989bp) DNA sequences. Individuals also were characterized by karyotype and with geometric morphometric analyses of mandibles and skulls. Two hundred fourteen individuals from 91 localities were studied. All the data support the recognition of a new species in SW Anatolia: the Taurus ground squirrel Spermophilus taurensis sp. nov. The new species has a small distribution in the Taurus Mountains in an area that is a hotspot for biodiversity. Molecular clock analysis suggests that the new species diverged from the European ground squirrel, Spermophilus citellus, about 2.5 million years ago and that the ancestor of these two species diverged from the widespread Anatolian ground squirrel, Spermophilus xanthoprymnus, about 5 million years ago. Morphometric differentiation in skull and mandible shape among the three species is incomplete, but statistically significant. S. xanthoprymnus is subdivided into five cytochrome b phylogroups and we use these data to infer the location of glacial refugia where the species lived during the last glacial maximum. This study illustrates the potential of combined molecular and morphometric studies to uncover new Anatolian species and to reconstruct their phylogeographic history. The new species is important for squirrel taxonomy and for understanding Eurasian mammal evolution.     

达乌尔黄鼠实验室饲养、繁殖及其冬眠阵

为探索实验室条件下达乌尔黄鼠饲养与繁殖的方法及冬眠阵的发生规律,参照野生黄鼠冬眠洞穴的主要生态环境参数,建立人工冬眠屋,采用传统锯末技术记录冬眠阵。结果显示: (1) 处于春季繁殖期的黄鼠应以大鼠饲料为主,辅以少量黄瓜等,夏季活跃期交叉饲喂大鼠饲料与兔饲料,辅以多水的瓜果蔬菜,秋季育肥期以大鼠饲料为主,辅以高脂肪高蛋白的花生、豆类等。(2)雌鼠怀孕期为28 d 左右,哺乳期约一个月,雌鼠每窝产仔4 ～ 8 只,平均5.52 只;初生幼鼠两周内忌换垫料,并避免将异味带入鼠房。(3)黄鼠冬眠期从当年11月下旬至次年3 月上旬,平均93.95 d;冬眠阵睡眠时长平均7. 44 d,阵间激醒时长平均1.36 d,睡眠天数占整个冬眠期的89.9% ;整个冬眠期,黄鼠冬眠阵平均7. 55 个。(4)2009 年秋至2011 年春季,自野外共捕回黄鼠185 只, 存活146 只,存活率78. 9% 。在2006、2009 和2011 年的黄鼠繁殖期,共配对25 对,产仔138 只,成活92 只,成活率为66.7% 。结果表明,野生达乌尔黄鼠可在人工饲养条件下实现繁殖,并可在人工冬眠屋成功冬眠。     

Ca-ATPase activity and protein composition of sarcoplasmic reticulum membranes isolated from skeletal muscles of typical hibernator,the ground squirrel Spermophilus undulatus

Ca-ATPase activity in sarcoplasmic reticulum (SR) membranes isolated from skeletal muscles of the typical hibernator, the ground squirrel Spermophilus undulatus, is about 2-fold lower than that in SR membranes of rats and rabbits and is further decreased 2-fold during hibernation. The use of carbocyanine anionic dye Stains-All has revealed that Ca-binding proteins of SR membranes, histidine-rich Ca-binding protein and sarcalumenin, in ground squirrel, rat, and rabbit SR have different electrophoretic mobility corresponding to apparent molecular masses 165, 155, and 170 kDa and 130, 145, and 160 kDa, respectively; the electrophoretic mobility of calsequestrin (63 kDa) is the same in all preparations. The content of these Ca-binding proteins in SR membranes of the ground squirrels is decreased 3–4 fold and the content of 55, 30, and 22 kDa proteins is significantly increased during hibernation.     

Dwindling genetic diversity in European ground squirrels?

The European ground squirrel (Spermophilus citellus) is endangered and in decline. Populations are increasingly fragmented, and only a coordinated conservation effort at the European level may guarantee its long-term survival. To obtain a general population genetic picture on a larger geographic scale, we screened 117 individuals from seven local populations in Hungary, Romania, and Austria for allelic variation at eleven microsatellite loci. We found a high (23.4%) proportion of private alleles, and a moderate to somewhat elevated level (15.27%) of partitioning of genetic diversity among populations, compared to that found in many other terrestrial mammals. Genetic variability was significantly higher than in earlier studied Czech populations that are considered genetically depleted, but significantly lower than in undisturbed populations of S. suslicus and S. brunneus, that are similar to the European ground squirrel in their ecological requirements, reproductive biology, and social organization. Genetic diversity was also lower than in most presumably “undisturbed” populations of other Sciurid species. This, together with the observed level and pattern of genetic differentiation among populations, such as no significant increase of genetic differentiation with geographic distance and similar variance of genetic differentiation between populations independent of geographic distance, indicated the prevalence of relatively strong drift effects for all populations. A Bayesian STRUCTURE analysis and a factorial correspondence analysis concordantly revealed a fairly complex genetic composition of local populations, but no major geographic trend in the pattern of the genetic composition. Overall, the results suggest disintegration of local colonies that might earlier have been more connected genetically. The STRUCTURE analysis also suggested anthropogenic translocations among single Hungarian populations. Our data on genetic diversity and its distribution do not object to such conservation measures. Translocation of individuals particularly from nearby populations may increase the chances of survival of small and isolated populations and counteract inbreeding at low densities.     

Genetic and morphologic variability of annual glassworts (Salicornia L.) from the Gulf of Trieste (Northern Adriatic)

The genetic variability of four pre-determined morphotypes of Salicornia (S. patula, S. emerici, S. veneta and the “saline type”) from 10 locations on the Gulf of Trieste coast were studied by means of ploidy level estimation using flow cytometry and by molecular DNA analysis of ITS regions of nrDNA and cpDNA. Two groups, the diploids and tetraploids, with matching nrDNA sequences, were recognized. Two types of cpDNA emerged among the diploids; one the same as in tetraploids. This incongruence between nrDNA and cpDNA sequences indicates a hybridization with tetraploid maternal progenitors and demonstrates the evidence for reticulate evolution. The morphometry, based on generative morphological traits, did not clearly separate the four morphotypes. However, the most important characters—length of the middle fertile segment, length of the lateral flower, width of the scarious margin of the fertile segment in the floral region, conform to two genetically recognized types: diploid S. patula and the widely distributed tetraploid S. emerici, also comprising the “saline type” and morphotype, known as a charismatic endemic S. veneta, a flagship species for nature conservation. Other discriminative traits for diploid and tetraploid morphotypes are parameters of the flowers (comparison of length of the central vs. lateral flower) and stomatal index. The determination key is also given. The tetraploid S. emerici is by far the most common species of annual glassworts in the area, occupying more extreme habitats than a diploid S. patula, which mostly forms monodominate stands.     

Daily rhythmicity and hibernation in the Anatolian ground squirrel under natural and laboratory conditions

We studied daily rhythmicity of body temperature (T _b) before and during hibernation in Anatolian ground squirrels (Spermophilus xanthoprymnus) under natural and laboratory conditions using surgically implanted temperature loggers. Under both conditions, robust daily T _b rhythmicity with parameters comparable to those of other ground squirrel species was observed before but not during hibernation. Euthermic animals had robust daily T _b rhythms with a mean of 37.0°C and a range of excursion of approximately 4°C. No T _b rhythm was detected during torpor bouts, either because T _b rhythmicity was absent or because the daily range of excursion was smaller than 0.2°C. The general patterns of hibernation that we observed in Anatolian ground squirrels were similar to those previously observed by other investigators in other species of ground squirrels.     

Pollen tube reallocation in two preanthesis cleistogamous species, Ranalisma rostratum and Sagittaria guyanensis ssp. lappula (Alismataceae)

Pollen deposition on stigmas and pollen tube growth in two apocarpous species, Ranalisma rostratum and Sagittaria guyanensis ssp. lappula (Alismataceae), were examined with fluorescence microscopy. The reallocation of pollen tubes among pistils was observed in both species. The percentage of pollinated stigmas per flower was only 22.0% in R. rostratum and 51.0% in S. guyanensis, though the seed/ovule ratios are higher than 65% in both species. The number of pollen grains on each single stigma ranged from 0 to 96 in R. rostratum, and from 0 to 125 in S. guyanensis. When more than one pollen grain deposited on a stigma, all pollen tubes grew to the ovary, but only one of them turned towards the ovule and finally entered the nucleus. The other tubes grew through the receptacle tissue into ovules of adjacent carpels whose stigmas were unpollinated or pollinated later. The intercarpellary growth of pollen tubes could be a mechanism to increase the efficiency of sexual reproduction in an apocarpous gynoecium with low pollination on the pistils.     

Systematics and evolution of the genus Torrubiella (Hypocreales, Ascomycota)

Torrubiella is a genus of arthropod-pathogenic fungi that primarily attacks spiders and scale insects. Based on the morphology of the perithecia, asci, and ascospores, it is classified in Clavicipitaceae s. lat. (Hypocreales), and is considered a close relative of Cordyceps s. 1., which was recently reclassified into three families (Clavicipitaceae s. str., Cordycipitaceae, Ophiocordycipitaceae) and four genera (Cordyceps s. str, Elaphocordyceps, Metacordyceps, and Ophiocordyceps). Torrubiella is distinguished morphologically from Cordyceps s. lat. mainly by the production of superficial perithecia and the absence of a well-developed stipitate stroma. To test and refine evolutionary hypotheses regarding the placement of Torrubiella and its relationship to Cordyceps s. lat., a multi-gene phylogeny was constructed by conducting ML and Bayesian analyses. The monophyly of Torrubiella was rejected by these analyses with species of the genus present in Clavicipitaceae, Cordycipitaceae, and Ophiocordycipitaceae, and often intermixed among species of Cordyceps s. lat. The morphological characters traditionally used to define the genus are, therefore, not phylogenetically informative, with the stipitate stromata being gained and/or lost several times among clavicipitaceous fungi. Two new genera (Conoideocrella, Orbiocrella) are proposed to accommodate two separate lineages of torrubielloid fungi in the Clavicipitaceae s. str. In addition, one species is reclassified in Cordyceps s. str. and three are reclassified in Ophiocordyceps. The phylogenetic importance of anamorphic genera, host affiliation, and stipitate stromata is discussed.     

Genetic diversity and population divergence in fragmented habitats: Conservation of Idaho ground squirrels

The Idaho ground squirrel, which consists of a northern (Spermophilus brunneus brunneus) and a southern subspecies (S. b. endemicus), has suffered from habitat loss and fragmentation, resulting in a reduction in both numbers and geographic range of the species. The northern Idaho ground squirrel (NIDGS) is listed as a threatened subspecies under the Endangered Species Act, and the southern Idaho ground squirrel (SIDGS) is a candidate. Because Idaho ground squirrel populations are small and often isolated, they are susceptible to inbreeding and loss of genetic diversity through drift. This research evaluates levels of genetic diversity and patterns of population divergence in both subspecies of Idaho ground squirrels. We hypothesized that NIDGS would exhibit lower genetic diversity and greater population divergence due to a longer period of population isolation relative to most SIDGS populations. Genetic diversity and divergence were quantified using 8 microsatellite loci. Contrary to expectations, SIDGS populations exhibited consistently lower levels of microsatellite diversity. Additionally, NIDGS exhibited only modest divergence among populations, while divergence levels among SIDGS populations were highly varied. Preliminary evaluations of mitochondrial DNA diversity and structure revealed lower diversity in NIDGS and some differences in gene flow that warrant further study. Based on our results, we suggest different management strategies for the two subspecies. Habitat restoration appears to be the most desirable conservation strategy for NIDGS populations. In contrast, low genetic diversity observed in SIDGS may warrant supplementation of isolated populations through translocations or captive breeding to mitigate further loss of genetic variability.     

Definition of a novel symbiovar (sv. retamae) within Bradyrhizobium retamae sp. nov., nodulating Retama sphaerocarpa and Retama monosperma

In this paper we analyze through a polyphasic approach several Bradyrhizobium strains isolated in Spain and Morocco from root nodules of Retama sphaerocarpa and Retama monosperma. All the strains have identical 16S rRNA genes and their closest relative species is Bradyrhizobium lablabi CCBAU 23086^T, with 99.41% identity with respect to the strain Ro19^T. Despite the closeness of the 16S rRNA genes, the housekeeping genes recA, atpD and glnII were divergent in Ro19^T and B. lablabi CCBAU 23086^T, with identity values of 95.71%, 93.75% and 93.11%, respectively. These differences were congruent with DNA–DNA hybridization analysis that revealed an average of 35% relatedness between the novel species and B. lablabi CCBAU 23086^T. Also, differential phenotypic characteristics of the new species were found with respect to the already described species of Bradyrhizobium. Based on the genotypic and phenotypic data obtained in this study, we propose to classify the group of strains isolated from R. sphaerocarpa and R. monosperma as a novel species named Bradyrhizobium retamae sp. nov. (type strain Ro19^T = LMG 27393^T = CECT 8261^T). The analysis of symbiotic genes revealed that some of these strains constitute a new symbiovar within genus Bradyrhizobium for which we propose the name “retamae”, that mainly contains nodulating strains isolated from Retama species in different continents.     

Comparative analysis of Ca2+-signaling in brown preadipocytes of the ground squirrel (Spermophillus undulatus) and mouse

We compared the Ca²⁺ response to noradrenalin, tapsigargin, thimerosal, and ionomycin in brown preadipocytes of the ground squirrel (Spermophillus undulatus) and mouse. The ground-squirrel brown preadipocytes did not respond to noradrenalin in concentrations within the physiological range. Stimulation of the plasma membrane Ca²⁺-channels with thimerosal showed a considerable reduction of the calcium entry in cell precursors of the both species. Intracellular calcium stores liberated in the preadipocytes of the both species by tapsigargin and ionomycin in Ca²⁺-free medium were insignificant. Ca²⁺-entry in preadipocytes was not activated by the intracellular Ca²⁺-store depletion. The Ca²⁺ response of the ground squirrel brown preadipocytes was independent of an animal’s physiological state or annual seasons. Brown preadipocytes of ground squirrels (Spermophillus undulatus) may be considered to be high ionomycin-resistant cells with reduced Ca²⁺-signaling systems.