Mongolian Lark as an indicator of taxonomic,functional and phylogenetic diversity of steppe birds

Biodiversity is declining at an unprecedented rate.Many grassland species have been lost and are now of conservation concern.Identifying efficient biodiversity indicators is a key pillar of the global conservation strategy.Mongolian Lark(Melanocorypha mongolica) is a charismatic bird species abound in Mongolian steppes,and recent studies demonstrated that this species share similar habitat requirements an d likely co-occur with other grassland birds.We tested the hypothesis that Mongolian Lark would make a suitable indicator for different aspects of avian biodiversity.We made a large-scale bird survey in Inner Mongolia,and classified point counts into three groups based on different taxonomic,functional,and phylogenetic biodiversity measures.We identified species indicators of each group using the indicator value(IndVal) analysis,and examined the bird compositional difference among groups.Linear models were used for studying the patterns of occurrence of Mongolian Lark in relation to various biodiversity measures.Mon golian Lark showed the highest indicator value in comparison with other potential species indicators.Bird species richness is significantly higher at sites with Mongolian Lark,an d its occurrence was negatively correlated with species abundance,functional richness,Rao’s quadratic entropy and phylogenetic diversity.Mongolian Lark is therefore a suitable indicator species for avian biodiversity in Mongolian steppes.It is highly distinctive in flight and usually seen singly.The widespread distribution and charismatic appearance make it easy to monitor and adequate for citizen science,and may provide useful information on the paradigm of surrogacy in conservation ecology.

Comparison of Constituents and Insecticidal Activities of Essential Oil from Artemisia lavandulaefolia by Steam Distillation and Supercritical-CO_2 Fluid Extraction

Essential oil was extracted from Artemisia lavandulaefolia DC.by steam distillation（SD） and supercritical-CO2 fluid extraction（SFE）,respectively.The constituents of the essentil oils extracted with those two methods were analyzed by gas chromatography-mass spectrometry（GC-MS） and insecticidal activities of the essential oils were evaluated,then the results were compared to assess their biological activity.Thirty-one compounds were identified in the essential oil extracted by SD,and its main components were eucalyptol,α,α,4-trimethyl-3-cyclohexene-1-methanol and so on.Twenty-two compounds were identified for the essential oil extracted by SFE,and its main components were cyclodecene,n-hexadecanoic acid and so on.Six chemical compositions were all contained in the essential oils extracted by the two methods,i.e.,eucalyptol,α,α,4-trimethyl-3-cyclohexene-1-methanol,caryophyllene,[3aS-（3aα,3bβ,4β,7α,7aS）]-octahydro-7-methyl-3-methylene-4-（1-methylethyl）-1H-cyclopenta[1,3]cyclopropa-[1,2]benzene,nerolidol and（-）-Spathulenol.The fumigation toxicity of the essential oil obtained by means of SD to the adults of Sitophilus zeamais is significantly higher than that of the essential oil by means of SFE.The contact toxicity of the essential oil obtained by means of SFE to the adults of S.zeamais is higher than that of the essential oil obtained by means of SD,but the difference is not significant.

Function of note strings in Japanese Tit alarm calls to the Common Cuckoo:a playback experiment

Background: Birds produce alarm calls to convey information about threats. Some Passerine alarm calls consist of several note strings, but few studies have examined their function. Previous studies have shown that Japanese Tits(Parus minor) can alter the calling rate and number and combination of notes in response to predators. We previously found the combinations of note types in Japanese Tit alarm calls to be significantly different in response to the Sparrowhawk(Accipiter nisus) and Common Cuckoo(Cuculus canorus).Methods: Through playback experiments, we tested whether the note strings in Japanese Tit alarm calls to the Common Cuckoo have different functions in conveying information. The note strings of selected alarm calls were divided into the categories of C and D, and different calls were then constructed separately based on the two note string categories. Original alarm calls(C–D), C calls and D calls were played back to male Japanese Tits during the incubation period.Results: Male Japanese Tits had a significantly stronger response to C calls than to C–D calls, and they showed a significantly stronger response to both C and C–D calls than to D calls, suggesting that Japanese Tits discriminated between the C and D calls.Conclusions: Our study demonstrated that the C-and D-category note strings of Japanese Tit alarm calls to the Common Cuckoo have different functions, which supports the previous finding that different note strings in an alarm call can provide different information to receivers. However, the exact meanings of these note strings are not yet known, and further investigation is therefore required.

High level of extra-pair paternity in the socially monogamous Marsh Tits(Poecile palustris)

Background:Extra-pair copulation behavior has been widely studied among socially monogamous birds.Many species revealed high rates of extra-pair paternity.But few of the studies have been carried out in the Asian population.Methods:From 2012 to 2019,we explored the extra-pair paternity of Marsh Tits(Poecile palustris)in Xianrendong National Nature Reserve,Liaoning Province,China.During the study,adult Marsh Tits were captured with mist nets and parental birds,with nest-box traps.Blood samples were taken from the brachial vein.Parentage analyses were carried out using nine highly variable microsatellite loci through Cervus 3.0 software and maximum likelihood approach.Results:Forty-nine offspring(15.08%)from 20 nests(45.45%)were the results of extra-pair fertilization out of a total of 325 offspring in 44 nests.The average extra-pair offspring ratio was 33.54%,with a set varying from 11.11 to 71.43%.Nine extra-pair fathers had been successfully identified,four of whom were the close neighbors of the focus nest while the nests of the remaining five were relatively far.No significant difference was found in the genetic similarity between the social and extra-pair mates of the female,nor in the heterozygosis among the maternal half-siblings.Conclusions:In general,our study proved that the extra-pair paternity in Marsh Tits and its extra-pair mating is independent of the genetic compatibility hypothesis.This complements the understudied bird's extra-pair paternity in Asian area and contributes to the comprehensive insight of birds'extra-pair paternity behaviors.

Egg-spot matching in common cuckoo parasitism of the oriental reed warbler: effects of host nest availability and egg rejection

Background: The success of cuckoo parasitism is thought to depend largely on the extent of egg matching between cuckoo and host eggs, since poor-matching cuckoo egg would lead to more frequent egg rejection by the host. In this study, we investigated how egg-spot matching between the Common Cuckoo(Cuculus canorus) and its host, the Oriental Reed Warbler(Acrocephalus orientalis) is affected by the local parasitism rate, nest availability in breeding synchronization and egg rejection.Methods: We used the paired design of parasitized and their nearest non-parasitized nests where breeding occurred simultaneously to compare egg-spot matching. The image analysis was used to compare four eggshell pattern variables, namely spot size, density, coverage on the different areas of egg surface, and the distribution on the whole egg surface. Egg recognition experiments were conducted to test the effect of egg spots on egg rejection by the host.Results: Our results show that much better matching in almost all spot parameters tested on the side of the egg and the spot distribution on the whole egg occurred in parasitized nests than in non-parasitized nests. Matching of spot density between cuckoo and host eggs in parasitized nests increased with the synchronization between temporal availability of nests and the egg-laying period of female cuckoos. Egg recognition experiments in which the warbler eggs were deliberately painted with extra spots led to a significantly higher egg rejection rate(78.3%) than of unpainted eggs.Conclusion: Our data suggest that both the high temporal encounter rate between cuckoo and warbler nests as well as the high egg rejection ability of the host are important factors for egg-spot matching of the cuckoos.

Egg recognition abilities of tit species in the Paridae family:do Indomalayan tits exhibit higher recognition than Palearctic tits?

DEAR EDITOR,Recent studies have shown that the closely related cinereous tit(Parus cinereus)and green-backed tit(P.monticolus)in China display strong egg recognition ability in contrast to tit species in Europe,which lack such ability.However,egg recognition in other populations of cinereous and greenbacked tits and additional Paridae species still requires further research.

Stereotypy and variability of social calls among clustering female big-footed myotis(Myotis macrodactylus)

Echolocating bats have developed advanced auditory perception systems, predominantly using acoustic signaling to communicate with each other They can emit a diverse range of social calls in complex behavioral contexts. This study examined the vocal repertoire of five pregnant big-footed myotis bats （Myotis macrodactylus）. In the process of clustering the last individual to return to the colony （LI） emitted social calls that correlated with behavior, as recorded on a PC-based digital recorder. These last individuals could emit 10 simple monosyllabic and 27 complex multisyllabic types of calls, constituting four types of syllables. The social calls were composed of highly stereotyped syllables, hierarchically organized by a common set of syllables. However, intra-specific variation was also found in the number of syllables, syllable order and patterns of syllable repetition across call renditions. Data were obtained to characterize the significant individual differences that existed in the maximum frequency and duration of calls. Time taken to return to the roost was negatively associated with the diversity of social calls. Our findings indicate that variability in social calls may be an effective strategy taken by individuals during reintegration into clusters of female M. macrodactylus.

Responses of soil Collembolans to vegetation restoration in temperate coniferous and broad-leaved mixed forests

A total of 900 soil samples were collected from five habitats,including primary coniferous broad-leaved mixed forests,secondary coniferous broad-leaved mixed forests,secondary broad-leaved forests,secondary shrub forests,and cutover lands in spring,summer,and autumn to quantify responses of soil Collembolans(springtails)to the restoration of vegetation of temperate coniferous and broad-leaved mixed forests.The results reveal that the taxonomic composition of Collembolans varied in the different stages of vegetation restoration.Seasonal variations were in regard to their abundance and richness.High similarities existed in Collembola communities at different stages of vegetation restoration,and distribution patterns of Collembola taxa displayed an evenness throughout all habitats.Soil Collembolans tended to gather on litter layers and soil surface;the highest abundance was found in the upper 5 cm soil layer during the initial stages of vegetation recovery.Tomocerus,Proisotoma,and Folsomia genera responded positively to the restoration of vegetation.However,responses of Ceratophysella and Parisotoma genera were negative.In addition,the Onychiuridae family did not respond to the vegetation restoration process.It was concluded that restoration of vegetative cover can increase the abundance of soil Collembolans,but different genera respond differently.

Identification of vocal individuality in male cuckoos using different analytical techniques

Background: Individuality in vocalizations may provide an effective tool for surveying populations of the Common Cuckoo(Cuculus canorus) but there remains few data on which technique to use to identify individuality. In this research, we compared the within-and between-individual variation in cuckoo calls using two different analytical methods, and discuss the feasibility of using call individuality to count male cuckoos within a population.Methods: We recorded vocalization from 13 males, and measured 15 spectro-temporal variables for each call. The majority of these call variables(n = 12) have greater variation between individuals than within individual. We first calculated the similarity(Pearson's R) for each paired calls in order to find a threshold that could distinguish calls emitted from the same or different males, and then counted the number of males based on this distinction. Second, we used the more widely accepted technique of discriminant function analysis(DFA) to identify individual male cuckoos, and compared the correct rate of classifying individuals between the two analytical methods.Results: Similarity of paired calls from the same male was significantly higher than from different males. Under a relatively broad threshold interval, we achieved a high(>90%) correct rate to distinguish calls and an accurate estimate of male numbers. Based on banded males(n = 3), we found the similarity of paired calls from different days was lower when compared with paired calls from the same day, but this change did not obscure individual identification, as similarity values of paired calls from different days were still larger than the threshold used to distinguish calls from the same or different males. DFA also yielded a high rate(91.9%) of correct classification of individuals.Conclusions: Our study suggests that identifying individual vocalizations can form the basis of an appropriate survey method for counting male cuckoos within a population, provided the performance of different analytical techniques are compared.

The spatial distribution and breeding behavior of neighbors affect the reproductive success of tits

The spatial distribution and breeding behavior of neighboring birds(conspecific and heterospecific)may influence reproduction and the effects differ across species.In this study,we investigated intra-and inter-specific spatial distribution of breeding via artificial nestboxes and examined whether overlap with neighbors affected reproductive success of the focal breeding pair.Regarding spatial distribution,Varied Tits(Sittiparus varius)and Cinereous Tits(Parus cinereus)showed similar pattern:distances of nestboxes containing conspecific tits were greater than those with heterospecific ones.In terms of breeding behavior,reproductive success was not significantly related to the distance from their neighbor’s nest.The reproductive success of Varied Tits was significantly related to that of their neighbors,while in Cinereous Tits,no strong link was found to any recorded reproductive behavior of the neighbors.These findings explain spatial variation in nest site selection of Varied Tits and Cinereous Tits.Reproductive success rate of Varied Tits is affected by that of the neighbors while that of Cinereous Tits is not.The results highlight that there are differences in reproductive outcome among related species and offer suggestions on the use of artificial nestboxes in experiment and conservation programs.

Plasma levels of luteinizing hormone and prolactin in relation to double brooding in Great Tit(Parus major)

The reproductive behaviors of birds are mainly controlled by the hypothalamus-pituitary-gonad axis.Many studies have shown that reproductive hormones are tightly linked to the breeding sub-stages.However,only a few studies have examined the temporal trend of hormone levels among different reproductive stages in multiple brooded species.We investigated the changes in plasma luteinizing hormone(LH)and prolactin(PRL)concentrations during different reproductive stages of the facultative double-brooded Great Tit(Parus major).We found that the concentrations of LH and PRL in females were significantly higher than those in males.Females had significantly higher LH and lower PRL concentrations in the pre-breeding period than in the first/second brooding periods,and there were no significant changes between the first and second brooding periods.The concentrations of LH and PRL in males had no significant difference between the pre-breeding period and the first brooding periods,while LH and PRL concentrations in the second brooding period were significantly higher than those in the first brooding period.We conclude that there are sex-based differences between LH and PRL at different stages of reproduction.The changes in LH and PRL in both males and females should be related to their physiological functions.Especially for males,individuals with higher levels of LH and PRL are more likely to maintain second clutches.

Plastering mud around the entrance hole affects the estimation of threat levels from nest predators in Eurasian Nuthatches

Background:For cavity-nesting birds,the nest entrance plays an important role in preventing predators from accessing nests.Several species of nuthatches use mud to narrow the entrance of cavities.In theory,the smaller the entrance hole size,the more effective it is against predators;however,few studies have tested whether narrowing the entrance hole size can affect the estimation of threat levels from nest predators in cavity-nesting birds.Methods:Using dummy experiments,we tested whether Eurasian Nuthatches(Sitta europaea,narrow the entrance hole of cavities)and Cinereous Tits(Parus cinereus,do not narrow the entrance hole,as a control)perform different nest defence behaviours against Common Chipmunks(Tamias sibiricus,small nest predator)and Red Squirrels(Sciurus vulgaris,larger nest predator).Results:Both nuthatches and tits exhibited stronger response behaviours(high dummy response scores)against chipmunks than against squirrels.Compared with tits,nuthatches exhibited more aggressive behaviours to chipmunks,but their responses to squirrels were similar.Conclusions:Nest defence behaviours of nuthatches to chipmunks differed from tits,and the results suggested that nuthatches might estimate threat levels of nest predators according to their narrowed entrance-hole size.

Newly Discovered Virus Associated Enzyme Capable of Alteration of Nucleic Acid Structures through Phosphotriester/-Diester Bonds

A newly discovered enzyme, that can catalyze the formation of phosphotriester/-diester bonds between nucleic acids, was found to be associated with plant and animal viruses; （i.e. southern bean mosaic virus, brome mosaic virus, influenza virus, avian virus and mouse retrovirus）. A partially purified enzyme from maize developing endosperms was prepared through 15%-35% ammonium sulfate fractionation, DEAE-cellulose anion exchange column chromatography and Sephadex GI50 gel filtration. The enzyme preparation was then used to demonstrate its main functional characteristics. The enzyme can use varieties of short and long chain length of nucleotides as substrates. However, the enzyme requires at least a minimum of 3 to 4 units of nucleotide chain length for the reaction to occur. The enzyme activity shows an optimum reaction in 50 mM sodium acetate buffer at pH 5.4 and is significantly inhibited by 6-azauridine as compared to other nucleotide analogs. By analyzing the data documented in literature and the results from the present study of the association of this enzyme with viruses and the distinctive inhibitory effect of 6-azauridine, it is speculated that this enzyme is likely associated with many other plant and animal viruses. The association of this enzyme on the surface of virus particles can be explored as a common antigen for developing a versatile antiviral vaccine.

Identification and functional characterization of female antennae-biased odorant receptor 23 involved in acetophenone detection of the Indian meal moth Plodia interpunctella

The Indian meal moth,Plodia interpunctella(Lepidoptera:Pyralidae),a globally distributed storage pest,relies on odors that are emitted from stored foods to select a suitable substrate for oviposition.However,the molecular mechanism underlying the chemical communication between P.interpunctella and its host remains elusive.In this study,130 chemosensory genes were identified from the transcriptomes of 7 P.interpunctella tissues,and the quantitative expression levels of all 56 P.interpunctella odorant receptor genes(PintORs)were validated using real-time quantitative polymerase chain reaction.The functional characteristics of 5 PintORs with female antennae-biased expression were investigated using 2-electrode voltage clamp recordings in Xenopus laevis oocytes.PintOR23 was found to be specifically tuned to acetophenone.Acetophenone could elicit a significant electrophysiological response and only attracted mated females when compared with males and virgin females.In addition,molecular docking predicted that the hydrogen bonding sites,TRP-335 and ALA-167,might play key roles in the binding of PintOR23 to acetophenone.Our study provides valuable insights into the olfactory mechanism of oviposition substrate detection and localization in P.interpunctella and points toward the possibility of developing eco-friendly odorant agents to control pests of stored products.

Social experience promotes the exploratory behaviour in Java Sparrows

Personality widely exists in diverse animal taxa.Such inter-individual variance in behaviour is supposed to be influenced by social context.However,it remains unknown whether the experience of social life has any carryover effects on the subsequent expression of personality.Here,we examined exploratory behaviour in caged Java Sparrows(Lonchura oryzivora)using exploration assays.Birds were assigned to live in either a solitary or a social context for four weeks.We compared the expressions of exploration before and after the treatments,and found that birds showed higher exploration tendencies after than before social life,while the isolated birds were consistent in their exploratory behaviours.Different living experience led to differences in the exploration activities for birds without significant differences in exploration before.Our results indicate that social experience can make birds more proactive.

Glomerular organization in the antennal lobe of the oriental armyworm Mythimna separata

In insects,the number and specificity of antennal lobe glomeruli often reflect the number and diversity of expressed chemosensory genes,which are linked to its ecological niche and specific olfactory needs.The oriental armyworm,Mythimna separata,is an important and common lepidopteran pest of cereal crops.Given its reliance on the olfactory system for crucial behaviors,understanding the evolutionary potential of this system requires a thorough characterization of the anatomical structure of the primary olfactory center.Here,we systematically identified all antennal lobe glomeruli of M.separata based on synaptic antibody immunostaining and mass staining of the olfactory sensory neurons.A total of 69 glomeruli were identified in females and 65 in males,and an intuitive nomenclature based on glomerular positions was applied.Our findings uncovered some sex-specific glomeruli in this species.There were ten female-specific glomeruli and three male-specific glomeruli,except for the macroglomerular complex(MGC)units,with a notable observation that the female labial pit organ glomerulus was larger than its male counterpart.Additionally,we identified four antennal-lobe tracts(ALTs)and retrograde labeling from the calyx revealed that all glomeruli were innervated by the medial ALT projection neurons.The comparison of the olfactory system structures between M.separata and sympatric moths supports their evolutionary convergence in noctuid moths.These results collectively lay the foundation for future studies on olfactory processing in M.separata.

Cannot see you but can hear you: vocal identity recognition in microbats

Identity recognition is one of the most critical social behaviours in a variety of animal species. Microchiropteran bats present a special use case of acoustic communication in the dark. These bats use echolocation pulses for navigating, foraging, and communicating; however, increasing evidence suggests that echolocation pulses also serve as a means of social communication. Compared with echolocation signals, communication calls in bats have rather complex structures and differ substantially by social context. Bat acoustic signals vary broadly in spectrotemporal space among individuals, sexes, colonies and species. This type of information can be gathered from families of vocalizations based on voice characteristics. In this review we summarize the current studies regarding vocal identity recognition in microbats. We also provide recommendations and directions for further work.

Egg recognition in Cinereous Tits(Parus cinereus):eggshell spots matter

Background:Brood parasitic birds such as cuckoos(Cuculus spp.)can reduce their host’s reproductive success.Such selection pressure on the hosts has driven the evolution of defense behaviors such as egg rejection against cuckoo parasitism.Studies have shown that Cinereous Tits(Parus cinereus)in China have a good ability for recognizing foreign eggs.However,it is unclear whether egg spots play a role in egg recognition.The aims of our study were to investigate the egg recognition ability of two Cinereous Tit populations in China and to explore the role of spots in egg recognition.Methods:To test the effect of eggshell spots on egg recognition,pure white eggs of the White-rumped Munia(Lonchura striata)and eggs of White-rumped Munia painted with red brown spots were used to simulate experimental parasitism.Results:Egg experiments showed that Cinereous Tits rejected 51.5%of pure white eggs of the White-rumped Munia,but only 14.3%of spotted eggs of the White-rumped Munia.There was a significant difference in egg recognition and rejection rate between the two egg types.Conclusions:We conclude that eggshell spots on Cinereous Tit eggs had a signaling function and may be essential to tits for recognizing and rejecting parasitic eggs.

Saltmarsh vegetation and social environment influence flexible seasonal vigilance strategies for two sympatric migratory curlew species in adjacent coastal habitats

Background:Animals need to adjust their vigilance strategies when foraging between physically contrasting veg-etated and non-vegetated habitats.Vegetated habitats may pose a greater risk for some if vegetation characteristics function as a visual obstruction but benefit others if they serve as protective shelter.Variation in group size,presence of similar species,along with variation in environmental conditions and anthropogenic disturbance can also influence vigilance investment.Methods:In this study,we quantified the vigilance behaviour of two large-bodied,sympatric migratory curlew species-Far Eastern Curlew(Numenius madagascariensis)and Eurasian Curlew(N.arquata)-in vegetated Suaeda salsa saltmarsh and non-vegetated mudflat habitat in Liaohekou National Nature Reserve,China.We used linear mixed models to examine the effects of habitat type,season,tide time,flock size(conspecific and heterospecific),and human disturbance on curlew vigilance investment.Results:Both species spent a higher percentage of time under visual obstruction in S.salsa habitat compared to mudflat habitat but in response,only Far Eastern Curlew increased their percentage of vigilance time,indicating that visual obstruction in this habitat is only a concern for this species.There was no evidence that S.salsa vegetation served as a form of cryptic background colouration since neither species decreased their vigilance effect in S.salsa habitat in spring compared to the autumn migration season.The effect of curlew social environment(i.e.flock size)was habitat dependent since percentage of vigilance time by curlews in saltmarsh increased with both the number of individual curlews and number of other birds present,but not in mudflat habitat.Conclusions:We conclude that both migratory curlew species exhibit a flexible vigilance adjustment strategy to cope with the different environmental and social conditions of adjacent and sharply contrasting coastal habitats,and that the trade-off between the risks of foraging and the abundance of prey may be a relatively common phenom-enon in these and other shorebird populations.

Old nest material functions as an informative cue in making nest-site selection decisions in the European Kestrel(Falco tinnunculus)

Background:For secondary cavity-nesting bird species that do not add lining materials to nests,the presence of old nest material or organic remains that have accumulated within nest cavities from previous breeding events may be a cue of nest-site quality.These materials potentially contain information about past breeding success in con-and heterospecifics and may improve the thermal insulation of eggs during incubation.However,few studies have addressed whether the presence of old nest materials serves as a cue for cavity-nesting raptors when choosing specific nest sites.Methods:We conducted a 9-year nest box experiment to test whether old nest materials from con-and heterospecifics serve as informative cues to the European Kestrel(Falco tinnunculus)when making nest selection decisions,as this species uses nest boxes without adding nesting material.Results:The presence of old nest materials and entrance size best discriminated nest boxes occupied by European Kestrels from unoccupied boxes.Nest boxes containing conspecific organic remains,artificial dry leaf and branch material,and material left behind by Great Tits(Parus major)were reused at higher rates,especially those containing conspecific nest material,than nest boxes containing true or simulated nest materials from predators.In 2010,no single nest box was occupied by the same banded individual that occupied the box in the previous year(10 females and 2 males were banded in 2009).Conclusions:European Kestrels preferred nest boxes containing old nest material over empty boxes,which is consistent with previous findings that they exploit con-and heterospecific cues when deciding where to settle and breed,as old nest or organic material provides substrate for incubating females.Kestrels may be able to assess the predation risks associated with a specific nest site based on experience or the presence of prey remains.The repeated use of nest boxes across breeding seasons by kestrels cannot be entirely ascribed to philopatry.This study provides evidence that old nest materials are potentially used as informative cues when making nest-site selection decisions in European Kestrels.