Odorrana Tormota Research Articles

Goal-directed bodily signals in birds and frogs.

Researchers have recently described the wing-fluttering signal of Japanese tits and eyeblink signal of concave-eared torrent frogs as bodily communication that elicits specific responses. I assess the evidence that these may be intentional, goal-directed signals using established criteria for gestural communication.

Female frogs communicate with males through blinking

Blinking is a behavior unique to animal taxa with motile eyelids, such as most amphibians and reptiles as well as all birds and mammals1. Eyelid movement has physiological functions, such as lubricating the cornea and washing away dust, but its potential signaling functions are not well understood1,2. The use of eyeblinks as a social signal is currently thought to be restricted to some primates, especially humans and their companion animals, but has not been verified in other taxa1,3,4. Here, through field observation and experiments, we demonstrate that female concave-eared torrent frogs (Odorrana tormota), which inhabit noisy streams, use eyeblinks to communicate with males to urge them to initiate amplexus for mating. Our findings reveal that eyeblinks may serve as a social signal in non-primate species.

Frequency jumps and subharmonic components in calls of female Odorrana tormota differentially affect the vocal behaviors of male frogs

Studies have demonstrated that the sounds of animals from many taxa with nonlinear phenomena (NLP)—caused by nonlinear characteristics of vocal organ dynamics that lead to nonlinear vocal phenomena—can influence the behavior of receivers. However, the specific functions of different NLP components have received less attention. In most frog species, females produce few or no vocalizations; in contrast, female Odorrana tormota exhibit a diverse range of calls that are rich in NLP components. Previous field playbacks have shown that the female calls can elicit responses from male frogs. Therefore, we conducted a phonotaxis experiment to investigate the differential effects of different NLP calls by female O. tormota on the vocal behavior of male frogs. The results revealed that calls with subharmonics elicited a greater number of short calls and answering calls from male frogs compared to calls with frequency jumps. However, calls with frequency jumps triggered more staccato calls from males than calls with subharmonics. Additionally, during the phonotaxis experiments, we recorded the initial vocalizations of males in response to playbacks of female calls. The majority of males first produced short calls. Under calls with frequency jumps, most of male frogs approaching within 10 cm of the loudspeaker produced staccato calls instead of “meow” calls or short calls. While under calls with subharmonics, most male frogs preferred to produced short calls. Our findings demonstrate that frequency jumps and subharmonic components in the calls of female O. tormota have different effects on male vocal behaviors. The current study lays a foundation for a further understanding of the function of anuran NLP components.

The succession of gut microbiota in the concave-eared torrent frog (Odorrana tormota) throughout developmental history.

The gut microbiota of amphibians plays a crucial role in maintaining health and adapting to various developmental stages. The composition of gut microbial community is influenced by the phylogeny, habitat, diet, and developmental stage of the host. The present study analyzed the microbiota in the intestine of O. tormota at 11 developmental stages (from the tadpole at Gosner stage 24 to the 3-year-old adult) using high-throughput 16S rRNA sequencing. Alpha diversity index analysis of the microbiota revealed that the index decreased from tadpole at Gosner stage 24 to adult frog stage, remained stable during the adult frog stages, but increased significantly at the early metamorphosis and hibernation preparation stages. The gut microbiota structure is similar in adult frogs but differs significantly in other developmental stages. Furthermore, the dominant phyla of gut microbiota in tadpoles were Proteobacteria, Firmicutes, Actinobacteria, and Bacteroidetes, whereas those in adult frogs were Proteobacteria, Firmicutes, Bacteroidetes, and Verrucomicrobia. Host and environmental factors jointly affected the gut microbial diversity and community composition of O. tormota, but developmental stage, feeding habit, and habitat type had a more significant influence. The microbial community in the gut varies with the developmental stage of the host and constantly adapts to the survival requirements of the host. These findings advance our understanding of the evolutionary mechanism of amphibian gut microbiota in maintaining health homeostasis and adaptation.

Optimal efficacy in mosquito-host interaction inhibition by 35-60 kHz animal sounds for malaria control

Female Anopheles gambiae sensu stricto, the malaria vector, continues to develop resistance to synthetic insecticides thus threatening its control. Studies on mosquitoe's phonotaxis showed that the electronic mosquito repellent (EMR) (40-55 kHz) sound, Odorrana tormota (35-60 kHz) sound and winded-EMR sound yielded 68.99%, 45.88% and 60.70% repellency respectively, supporting feasibility of non-pollutant sounds in mosquito control. Rudimental studies with O. tormota sounds didn’t observe protocols for mosquito rearing standard operation procedures (SOPs) and World Health Organisation (WHO) guidelines for efficacy testing of spatial repellents. Also, phonotactic inhibition by natural sounds of Delphinapterus leucas and male A. gambiae s.s had not been studied. This research, therefore, determined spatial activity index (SAI) and protection index (PI) of mated female A. gambiae s.s evoked by 35-60 kHz sounds of O. tormota, D. leucas, and male A. gambiae s.s in adherence to mosquito rearing SOPs and WHO guidelines. Mosquitoes were reared in Kenya Medical Research Institute (KEMRI) laboratories. Sounds of a hundred 3-5-day-old male A. gambiae s.s were recorded using Avisoft recorder whereas the sounds of O. tormota and D. leucas were acquired. Abnormal mosquito behavioural responses were observed during the bioassays. The bioassay data was collected and analyzed using statistical softwares. Mean SAI evoked by O. tormota, male A. gambiae s.s, and D. leucas were 0.142, 0.318, and 0.206 respectively, indicating negative phonotaxis. The PI elicited by O. tormota was 80.06% exceeding male A. gambiae s.s and D. leucas by 1.65% and 8.24% respectively. The constant frequency modulated sound of O. tormota yielded mean bandwidth (mean entire; 13.95 kHz), maximum frequency (mean entire; 41.61 kHz), maximum energy (8.02 Pa2s), entropy (2.87 bits), delta power (24.10 dB), average power (55.60 dB), maximum power frequency (38.09 kHz) and maximum power (75.90 dB).

Changes in vocal characteristics of male concave-eared torrent frogs (<i>Odorrana tormota</i>) in different chorus tides

Changes in vocal characteristics of male concave-eared torrent frogs (<i>Odorrana tormota</i>) in different chorus tides

DPOAEs and tympanal membrane vibrations reveal adaptations of the sexually dimorphic ear of the concave-eared torrent frog, Odorrana tormota

While most anuran species are highly vocal, few of them seem to be endowed with a complex call repertoire. Odorrana tormota, combines a remarkable vocalization complexity with auditory sensitivity over an extended spectral range spanning from audible to ultrasonic frequencies. This species is also exceptional for its ability to modify its middle ear tuning by closing the Eustachian tubes (ET). Using scanning laser Doppler vibrometry, the tympanal vibrations were measured to investigate if the tuning shift caused by the ET closure contributes to intraspecific acoustic communication. To gain insight into the inner ear frequency selectivity and sensitivity of this species, distortion product otoacoustic emissions were recorded at multiple frequency-level combinations. Our measurements of inner ear responses indicated that in O. tormota each sex is more sensitive to the frequencies of the other sex's vocalizations, female ears are more sensitive to 2–7 kHz, while male ears are more sensitive to 3–15 kHz. We also found that in both sexes the ET closure impacts the sensitivity of the middle and inner ear at frequencies used for communication with conspecifics. This study broadens our understanding of peripheral auditory mechanisms contributing to intraspecific acoustic communication in anurans.

Male antiphonal calls and phonotaxis evoked by female courtship calls in the large odorous frog (Odorrana graminea).

Acoustic communication plays a vital role in frog reproduction. In most anuran species, long-distance sound communication is one-way from males to females; during the reproductive season, males produce species-specific advertisement calls to attract gravid females, and females are generally silent but perform phonotactic movements that lead to amplexus. One exception is the concave-eared torrent frog (Odorrana tormota). In this species, females produce courtship calls that elicit antiphonal vocalizations by males, followed by precise phonotactic movements. The large odorous frog O. graminea (previously Odorrana livida) in southern China is subject to the same environmental constraints as O. tormota, with which it is sympatric; it is unclear whether their sound communication is one-way or bidirectional. Here, we provide the first data on female O. graminea vocalizations and their functions. Using playbacks of female calls, we conducted acoustic behavioral experiments in the laboratory in response to which males emitted single- or multi-note antiphonal calls with a varying fundamental frequency. Moreover, they were attracted to female call playbacks, exhibiting precise phonotaxis. The female courtship call-male response interaction thus forms a duet between partners of a receptive pair. These results demonstrate that this unique communication system likely reflects an adaptation to an environment in which short-distance communication is at a premium given the high levels of ambient noise.

Comparative transcriptome analysis provides insights into the molecular mechanisms of high-frequency hearing differences between the sexes of Odorrana tormota

BackgroundAcoustic communication is important for the survival and reproduction of anurans and masking background noise is a critical factor for their effective acoustic communication. Males of the concave-eared frog (Odorrana tormota) have evolved an ultrasonic communication capacity to avoid masking by the widespread background noise of local fast-flowing streams, whereas females exhibit no ultrasonic sensitivity. However, the molecular mechanisms underlying the high-frequency hearing differences between the sexes of O. tormota are still poorly understood.ResultsIn this study, we sequenced the brain transcriptomes of male and female O. tormota, and compared their differential gene expression. A total of 4,605 differentially expressed genes (DEGs) between the sexes of O. tormota were identified and eleven of them were related to auditory based on the annotation and enrichment analysis. Most of these DEGs in males showed a higher expression trend than females in both quantity and expression quantity. The highly expressed genes in males were relatively concentrated in neurogenesis, signal transduction, ion transport and energy metabolism, whereas the up-expressed genes in females were mainly related to the growth and development regulation of specific auditory cells.ConclusionsThe transcriptome of male and female O. tormota has been sequenced and de novo assembled, which will provide gene reference for further genomic studies. In addition, this is the first research to reveal the molecular mechanisms of sex differences in ultrasonic hearing between the sexes of O. tormota and will provide new insights into the genetic basis of the auditory adaptation in amphibians during their transition from water to land.

Comparative study on gut microbiota in three Anura frogs from a mountain stream.

Composition and diversity in gut microbiota are impacted by a wide variety of factors. The similarity of gut microbiota in related or sympatric species has been gaining recent traction. Here, 16S rRNA gene sequencing technology was employed to study the gut microbiota of three sympatric frog species, namely Odorrana tormota, O. graminea, and Amolops wuyiensis. In these three frog species, the most abundant phylum was Proteobacteria, followed by Bacteroidetes, Verrucomicrobia, and Firmicutes. The most abundant family was Burkholderiaceae in three species. The most dominant genera were Burkholderia, Caballeronia, and Paraburkholderia with the highest relative abundance in O. tormota, O. graminea, and A. wuyiensis, respectively. No differences were observed in alpha diversity indexes among the three frog species. However, bacterial similarity of gut microbiota was significantly different between O. tormota and A. wuyiensis and between O. graminea and A. wuyiensis. Metabolism‐related gene function was predominantly enriched in the gut microbiota of the three evaluated frog species. From these findings, that the relative abundance of the gut microbiota and predicted gene functions differed in three species, we conclude that there were significant differences in the gut microbiota of the three species. Similar alpha diversity and interspecific bacterial similarity in the gut might be related to bacterial transmission among the three Anura frogs evaluated in this study.

Dynamic transcriptome and histomorphology analysis of developmental traits of hindlimb thigh muscle from Odorrana tormota and its adaptability to different life history stages

BackgroundSystematic studies on the development and adaptation of hindlimb muscles in anura amphibians are rare. Here, we integrated analysis of transcriptome and histomorphological data for the hindlimb thigh muscle of Odorrana tormota (concave-eared torrent frog) at different developmental stages, to uncover the developmental traits of hindlimb thigh muscle from O. tormota and its adaptability to different life history stages.ResultsThe development of hindlimb thigh muscle from O. tormota has the following characteristics. Before metamorphosis, myogenous cells proliferate and differentiate into myotubes, and form 11 muscle groups at G41; Primary myofibers and secondary myofibers appeared during metamorphosis; 11 muscle groups differentiated continuously to form myofibers, accompanied by myofibers hypertrophy after metamorphosis; During the growth process of O. tormota from G42 to G46, there were differences between the sexes in the muscle groups that differentiate into muscle fibers, indicating that there was sexual dimorphism in the hindlimb thigh muscles of O. tormota at the metamorphosis stages. Some genes and pathways related to growth, development, and movement ability of O. tormota at different developmental stages were obtained. In addition, some pathways associated with adaptation to metamorphosis and hibernation also were enriched. Furthermore, integrated analysis of the number of myofibers and transcriptome data suggested that myofibers of specific muscle groups in the hindlimbs may be degraded through lysosome and ubiquitin pathways to transform into energy metabolism and other energy-related substances to meet the physiological needs of hibernation.ConclusionsThese results provide further understanding the hindlimb thigh muscle development pattern of frogs and their adaption to life history stages.

Female concave‐eared torrent frogs prefer smaller males

AbstractMate choice is a natural selection mechanism whereby the traits of select members of a population are passed on to their offspring. We hypothesized that, for stream‐dwelling anurans, females might prefer males with smaller body size and higher call frequency as their vocal calls are more conspicuous in the species’ habitat featuring intense but predominantly low‐frequency stream noise. To test this hypothesis, we first measured the body size of amplectant male concave‐eared torrent frogs (Odorrana tormota) in the field, a species with demonstrated ability of ultrasonic communication. Our field observation showed that, on average, amplectant males had smaller body size compared to that of non‐amplectant males (P < 0.05), an unusual finding among anurans. We next performed two‐choice amplexus experiments by pairing a female with two males in an aquarium indoors. Results of laboratory experiments similarly revealed female’s preference for males with smaller body sizes. Additionally, we recorded and analyzed the advertisement calls of 33 males and confirmed that their call frequency was inversely related to the body size as expected. Together, our studies showed that, for O. tormota, smaller males with higher call frequencies had higher mating success over bigger rivals, that is, females prefer smaller males.

Molecular characterization, expression analysis of myostatin gene and its negative regulation by miR-29b-3p in Chinese concave-eared frogs (Odorrana tormota).

The molecular characteristics, expression patterns and functions of the amphibian myostatin (MSTN) gene are unknown. Here, we isolated a full-length Odorrana tormota MSTN cDNA sequence of 1701 bp (Ot-MSTN), containing a putative N-terminal signal peptide, a TGF-β propeptide domain and an active peptide. Ot-MSTN was expressed in 9 selected tissues examined, and the highest level of expression was in thigh muscle, followed by brain and female gonadal tissue. The expression of Ot-MSTN in multiple O. tormota tissues supported that the activities of MSTN may be not limited to skeletal muscle. Ot-MSTN expression was decreased from stage 31 to stage 40, while the growth rate was increased. The expression of Ot-MSTN in adult male frogs increased with age, indicating that adult male frogs may inhibit the continued hypertrophy of thigh muscle fibers and decrease the growth rate of thigh muscle to ensure muscles do not grow too large. Luciferase reporter assays showed that miR-29b-3p directly targeted the 3'-UTR of Ot-MSTN. miR-29b-3p expression in the thigh muscle of 2 yrs. females who grew faster was significantly lower than that of the slow-growing 2 yrs. male individuals, which showed an opposite trend with Ot-MSTN expression. In addition，miR-29b-3p expression reversed trends of Ot-MSTN expression at different developmental stages in thigh muscle. Therefore, these data indicate that miR-29-3p may negatively regulate the expression of MSTN and regulate thigh muscle growth and development in O. tormota.

A Frog-Derived Immunomodulatory Peptide Promotes Cutaneous Wound Healing by Regulating Cellular Response.

Wound healing-promoting peptides exhibit excellent therapeutic potential in regenerative medicine. However, amphibian-derived wound healing-promoting peptides and their mechanism of action remain to be further elucidated. We hereby characterized a wound healing-promoting peptide, Ot-WHP, derived from Chinese concave-eared frog Odorrana tormota. It efficiently promoted wound healing in a mouse model of full-thickness wounds. Ot-WHP significantly increased the number of neutrophils in wounds, and modestly promoted neutrophil phagocytosis and phorbol myristate acetate (PMA)-induced neutrophil extracellular trap formation. Ot-WHP also significantly increased the number of macrophages in wound sites, and directly induced chemokine, cytokine and growth factor production in macrophages by activating mitogen-activated protein kinases (MAPKs) and nuclear factor-κB (NF-κB) signaling pathways. Of note, Ot-WHP did not act as a chemoattractant for neutrophils and macrophages, suggesting its chemotactic activity depends on inducing chemoattractant production in macrophages. Besides, Ot-WHP directly promoted keratinocyte migration by enhancing integrin expression and cell adhesion. In addition, Ot-WHP significantly enhanced the cross-talk between macrophages and keratinocytes/fibroblasts by promoting keratinocyte/fibroblast proliferation, and fibroblast-to-myofibroblast transition despite having no direct effects on keratinocyte/fibroblast proliferation, and fibroblast differentiation. Collectively, Ot-WHP directly elicited the production of regulatory factors in macrophages, consequently initiated and accelerated the inflammatory phase by recruiting neutrophils and macrophages to wounds, and in turn enhanced the cross-talk between macrophages and keratinocytes/fibroblasts, additionally promoted keratinocyte migration, and finally promoted cutaneous wound healing. Our findings provide a promising immunomodulator for acute wound management and new clues for understanding the mechanism of action of amphibian-derived wound healing-promoting peptides.

Effects of dispersal barriers and geographic distance on the genetic structure of a narrowly distributed frog in a spatially structured landscape

AbstractLandscape features (e.g., mountains and rivers) can act as barriers to dispersal and gene flow, and therefore impede population connectivity, increasing genetic differentiation between populations. The concave‐eared torrent frog (Odorrana tormota) is a rare, stream‐associated species in eastern China. In this study, we investigated the genetic structure and population demography of this narrowly distributed frog based on mitochondrial Cyt b gene and seven nuclear microsatellite loci. As a result, we found that the rare frog still preserved a relatively high level of genetic diversity compared to some other amphibia. Population structure analyses distinctly identified three or four tentative genetic clusters for the species in the study area. Additionally, by fine‐scale spatial autocorrelation analysis, significant positive genetic structure was uncovered in the shortest distance classes (0–5 km, 5–10 km). Demographic analyses revealed a population expansion (0.075–0.017 Mya) and 15 times population decline (c. 9000 years ago). In conclusion, we supposed that stable montane environments and associated historical population expansion might provide an opportunity for the species to harbor high genetic diversity. The relatively recent population decline might be correlated with climate change as well as genetic differentiation among populations. In addition, our results showed that, on a small landscape scale, dispersal is closely linked to geographic distance and the presence of river systems may not substantially affect the genetic structure for the narrowly distributed O. tormota.

Cover Image: Integrated mRNA and miRNA expression profile analyses reveal the potential roles of sex‐biased miRNA‐mRNA pairs in gonad tissues of the Chinese concave‐eared torrent frog (Odorrana tormota). J Exp Zool (Mol Dev Evol) 3–4/2019

Cover Image: Integrated mRNA and miRNA expression profile analyses reveal the potential roles of sex‐biased miRNA‐mRNA pairs in gonad tissues of the Chinese concave‐eared torrent frog (Odorrana tormota). J Exp Zool (Mol Dev Evol) 3–4/2019

Integrated mRNA and miRNA expression profile analyses reveal the potential roles of sex-biased miRNA-mRNA pairs in gonad tissues of the Chinese concave-eared torrent frog (Odorrana tormota).

The Chinese concave-eared torrent frog (Odorrana tormota) is typically sexually dimorphic. Females are significantly less common than males in the wild. Until now, the molecular mechanisms of reproduction and sex differentiation of frogs remain unclear. Here, we integrated mRNA and microRNA (miRNA) expression profiles to reveal the molecular mechanisms of reproduction and sex differentiation in O. tormota. We identified 234 differentially expressed miRNAs (DEMs) and 18,551 differentially expressed transcripts. Of these, 12,053 mRNAs and 64 miRNAs were upregulated in testes, and 6,498 mRNAs and 170 miRNAs were upregulated in ovaries. Integrated analysis of the miRNA and mRNA expression profiles predicted 75,602 potential miRNA-mRNA interaction sites, with 42,065 negative miRNA-mRNA interactions. We found 36 differentially expressed genes (DEGs) related to reproduction and sex differentiation, of which 15 DEGs formed 92 negative miRNA-mRNA interactions with 34 known DEMs. Thus, miRNAs may play other important roles in O. tormota. Furthermore, Gene Ontology enrichment and Kyoto Encyclopedia of Genes and Genomes pathway analyses showed reproductive-related processes, such as the gonadotropinreleasing hormone signaling pathway and ovarian steroidogenesis. Based on functional annotation and the literature, the retinoic acid signaling pathway, the SOX9-AMH pathway, and the process of spermatogenesis may be involved in the molecular mechanisms of reproduction and sex differentiation in O. tormota, and may be regulated by miRNAs. The miRNA-mRNA pairs described may provide further understanding of the regulatory mechanisms associated with reproduction and sex differentiation, and the molecular mechanism of reproduction in O. tormota.

Complete genome sequence of a divergent strain of Tibetan frog hepatitis B virus associated with a concave-eared torrent frog (Odorrana tormota).

Viruses of the family Hepadnaviridae are characterized by partially dsDNA circular genomes of approximately 3.2 kb, which are reverse transcribed from RNA intermediates. Hepadnaviruses have a broad host range, which includes humans (hepatitis B virus), other mammals (genus Orthohepadnavirus), and birds (genus Avihepadnavirus). The known host specificity of hepadnaviruses has been expanded by reports of new viruses infecting fish, amphibians, and reptiles. Tibetan frog hepatitis B virus (TFHBV) was recently discovered in a member of the species Nanorana parkeri (family Dicroglossidae) from Tibet. To increase our understanding of hepadnaviruses that infect amphibian hosts, we identified the full-length genome of a divergent strain, TFHBV-Ot, associated with a concave-eared torrent frog (Odorrana tormota, family Ranidae) from China by searching deep-sequencing data. TFHBV-Ot shared a genomic organization and 76.6% overall genome sequence nucleotide identity with the prototype TFHBV associated with N. parkeri (TFHBV-Np). The pairwise amino acid sequence identity between the predicted gene products of TFHBV-Ot and TFHBV-Np ranged between 63.9% and 77.9%. Multiple tissue/organ-specific RNAseq datasets suggested a broad tropism of TFHBV, including muscle, gonads and brain. In addition, we provide information about putative virus-derived small RNAs from an amphibian hepadnavirus. The results presented here expand the known genetic diversity and host range of TFHBV to Ranidae frogs, and warrant an investigation of hepadnaviral infection of amphibian brains.

High-Throughput Sequencing Analysis Reveals Correlations between Host Phylogeny, Gut Microbiota, and Habitat of Wild Frogs from a Mountainous Area

Gut microbiota of vertebrate hosts play crucial roles for the host they inhabit. In this study, we compared the composition and predicted functions of gut microbiota of female individuals belonging to four frog species from different habitats using bacterial 16S rRNA gene sequencing. Our results showed that the gut microbiota of the examined frog species were dominated by the phyla Firmicutes, Proteobacteria, Bacteriodetes, Verrucomicrobia, and Fusobacteria. There were significant differences in the relative abundance of dominant phyla among the four species, although the relative abundance of dominant phyla in the microbiota of individuals of Odorrana tormota was more evenly balanced (ranged from 10.45% for Fusobacteria to 27.21% for Bacteroidetes). The alpha diversity patterns varied depending on the microbial taxon level. At the phylum level, the Chao1 diversity was positively associated with body weight, with the diversity index scores of O. tormota and Amolops wuyiensis markedly lower than those of Odorrana schmackeri and Polypedates megacephalus. At the genus level, Chao1 diversity was associated with evolutionary relationships among species, with the Chao1 diversity index score for P. megacephalus of distant evolutionary relationships being significantly higher than the scores for frogs with close evolutionary relationships. Moreover, permutational multivariate analysis of variance revealed significant differences in the gut bacterial community structure among species (R2 = 0.510, P = 0.001). Further, the predicted functions of the gut microbiota were similar for species with close evolutionary relationships but differed with distant evolutionary relationships. Taken together, our results indicate that wild frogs have species-specific microbial communities and provide insight into the ecological and evolutionary dynamics of host–microbe interactions.

Analysis of the Acoustic Propagation Parameters of the Natural Sounds of Delphinapterus leucas and Odorrana tormota Fundamental in the Startle of the Female Anopheles Gambiae

The female Anopheles gambiae, a malaria vector detects ultrasound by the antennae causing startle responses attributed to neural stress. Resistance to the control and treatment measures by malaria vectors and pathogens has resulted to a low decline rate in malaria cases and deaths. Studies have shown a 46% evasive response in the A. gambiae by the 35-60 kHz sounds of Odorrana tormota. This study determined, analysed and compared acoustic propagation parameters of the sounds of Delphinapterus leucas and Odorrana tormota fundamental in the startle of the mosquito. The sound of D. leucas and O. tormota were acquired. The Avisoft-SASLab 5.2 and Raven Pro 1.5 software were used in the study. Spectral analysis of sounds of D. leucas and O. tormota showed composition of pulsate audible and utrasonic harmonics, FM and CF modulation. The mean of the fundamental frequency (mean entire) of D. leucas was 1.684kHz, less by 3.712kHz from that of the sound of O. tormota differing insignificantly. The sound of D. leucas recorded a maximum value of 64.8kHz in maximum frequency (mean) which was 25.5kHz above that of O. tormota with a highly significant difference. Also, the sound of D. leucas had a 64.10kHz bandwidth (mean entire) wider than that of O. tormota by 53.64kHz, a highly significant difference. The peak amplitude (mean entire) of the sound of D. leucas was 89.05Pa less by 39.837kHz from that of the sound of O. tormota differing significantly. The mean acoustic energy of D. leucas was 0.07Pa2s whereas the O. tormota possessed 2,496.63Pa2s differ insignificantly. These parameters are essential in the study of mosquito repellence and design of a repellent device.