Short Communication |
Corresponding author: Gang Cheng ( 25965716@qq.com ) Corresponding author: Gang Wei ( wg198553@126.com ) Academic editor: Günter Gollmann
© 2024 Jing Liu, Yanlin Cheng, Shize Li, Gang Cheng, Gang Wei.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation:
Liu J, Cheng Y, Li S, Cheng G, Wei G (2024) The advertisement call of Leptobrachella dushanensis (Anura, Megophryidae). Herpetozoa 37: 373-378. https://doi.org/10.3897/herpetozoa.37.e136688
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We describe the advertisement call of the Dushan Leaf Litter Toad, Leptobrachella dushanensis, based on recordings obtained from three individuals from its type locality. The advertisement call of L. dushanensis consists of a single note with a dominant frequency of 4.84–4.91 kHz. Furthermore, we compare the differences in advertisement calls between L. dushanensis and its sister species, Leptobrachella dong. The advertisement calls of L. dushanensis and L. dong can be clearly distinguished. Our findings provide bioacoustic evidence supporting the taxonomic status of L. dushanensis and provide basic data for future behavioral and ecological studies on Leptobrachella.
acoustic differences, bioacoustics, frogs, southern China
The Asian leaf litter toads of the genus Leptobrachella Smith, 1925, are the largest clade of the family Megophryidae Bonaparte, 1950, currently comprising 105 species (
Vocalizations of anuran amphibians have received much attention in studies of behavioral ecology and species identification (
Leptobrachella dushanensis (Anura, Megophryidae) is a newly described species in Southern China (
A total of 197 calls were recorded from three individuals of L. dushanensis, collected from its type locality Dushan County, Guizhou Province, China (107.63°E, 25.94°N, ca. 1200 m elev., 17.0–19.0 ℃ air temperature, 98% ambient humidity) on April 1, 2024, between 19:00–23:00 h. Additionally, to compare the differences in advertisement calls between L. dushanensis and its sister species, Leptobrachella dong, we also collected advertisement calls from L. dong. A total of 117 calls were recorded from four individuals of L. dong, collected from Congjiang County, Guizhou Province, China (108.28°E, 25.57°N, ca. 1190 m elev., 12.0–13.0 ℃ air temperature, 90% ambient humidity) on March 4, 2022, between 19:00–23:00 h. Calls from each individual were obtained using a digital recorder, a SONY ICD-PX470 (sampling rate 44.1 kHz, 16-bit resolution). Each call was recorded within a 0.5 m distance from the calling individual. All recordings were obtained from isolated individuals, without overlapping calls or mixed choruses, and saved as WAV files. The recordings are publicly available in Figshare at https://doi.org/10.6084/m9.figshare.26969512.
Snout-vent lengths (SVLs) of all recorded males were measured in situ using a precision digital caliper to the nearest 0.1 mm. One L. dushanensis (specimen number: DS20240401001) and one L. dong (specimen number: CJ20220304001) were collected for species identification, and the others were released to their original habitat after measurement. After taking photos of the two specimens, we euthanized them using isoflurane and then fixed the specimens in 10% buffered formalin. Tissue samples were taken and preserved separately in 95% ethanol before fixation. Specimens were deposited in the Moutai Institute, China. Mitochondrial COI genes were extracted and amplified from muscle samples and sequenced, and the obtained sequence was verified and uploaded to GenBank (https://www.ncbi.nlm.nih.gov/). The sequencing results were compared to finally determine the species collected.
The advertisement call characteristics were analyzed with the software Raven Pro 1.6 (K. Lisa Yang Center for Conservation Bioacoustics [at the Cornell Lab of Ornithology] 2023). Temporal properties were measured using Raven’s waveform display. Spectral properties were measured by averaging the spectrum over the entire duration of a call (Hann window, DFT = 512 samples, overlap = 50%, hop size = 256 samples). Only calls that had a high signal-to-noise ratio and were free from overlapping calls of nearby males were used for the analysis. We used “call-centred” terminology as summarised by
Males were observed calling on rocks in streams surrounded by shrubs. After molecular identification, we determined that the species captured in Dushan County is L. dushanensis (Fig.
Descriptive statistics for call parameters of advertisement calls of Leptobrachella dushanensis.
Individuals | Calls analyzed | Air temperature (°C) | SVL (mm) | Call duration (ms) | Inter-call interval (ms) | Call rate | Note per call | Note rate | No. of pulses per call | Pulse rate | Dominant frequency (kHz) |
---|---|---|---|---|---|---|---|---|---|---|---|
DS20220401001 | 75 | 17.0 | 31.9 | 123.78 ± 16.61 94.00–167.10 | 499.59 ± 49.57 410.80–594.31 | 1.78 ± 0.14 1.65–1.93 | 1 | NA | 21.00 ± 4.50 14.00–33.00 | 164.26 ± 21.72 134.30–223.40 | 4.91 ± 0.15 4.82–5.17 |
DS20240401002 | 77 | 19.0 | 32.3 | 86.76 ± 9.73 68.40–115.40 | 469.77 ± 77.65 303.27–669.00 | 2.09 ± 0.18 1.92–2.39 | 1 | NA | 15.00 ± 1.50 12.00–24.00 | 174.06 ± 12.40 152.01–205.72 | 4.87 ± 0.17 4.82–5.51 |
DS20240401003 | 45 | 19.0 | 32.6 | 78.19 ± 6.52 64.60–97.10 | 394.88 ± 68.53 288.00–588.02 | 2.31 ± 0.15 2.19–2.48 | 1 | NA | 14.00 ± 0.50 10.00–20.00 | 164.56 ± 12.13 139.32–195.67 | 4.84 ± 0.10 4.82–5.51 |
All Individuals | 3/197 | 17.0–19.0 | 31.9–32.6 | 78.19–123.78 | 394.88–499.59 | 1.78–2.31 | 1 | NA | 14.00–21.00 | 164.26–174.06 | 4.84–4.91 |
After molecular identification, we confirmed that the species collected from Congjiang County is L. dong (Fig.
Advertisement calls of the two Leptobrachella species. A. 5 s oscillograms showing nine calls of Leptobrachella dushanensis; B. 0.2 s oscillograms and corresponding spectrograms showing one call of L. dushanensis; C. 5 s oscillograms showing fourteen calls of Leptobrachella dong; D. 0.2 s oscillograms and corresponding spectrograms showing one call of L. dong.
Descriptive statistics for call parameters of advertisement calls of Leptobrachella dong.
Individuals | Calls analyzed | Air temperature (°C) | SVL (mm) | Call duration (ms) | Inter-call interval (ms) | Call rate | Note per call | Note rate | No. of pulses per call | Pulse rate | Dominant frequency (kHz) |
---|---|---|---|---|---|---|---|---|---|---|---|
CJ20220304001 | 27 | 13.0 | 29.6 | 82.55 ± 5.08 65.60–94.00 | 380.01 ± 42.06 312.95–478.13 | 2.23 | 1 | NA | 11.00 ± 0.00 8.00–13.00 | 121.87 ± 5.43 106.71–133.63 | 4.82 |
CJ20220304002 | 30 | 13.0 | 29.8 | 63.54 ± 4.84 48.40–72.80 | 336.30 ± 47.67 274.14–441.65 | 2.57 | 1 | NA | 9.00 ± 1.13 5.00–13.00 | 133.77 ± 21.62 70.80–165.41 | 4.82 |
CJ20220304003 | 30 | 12.0 | 29.5 | 90.82 ± 8.50 63.10–105.60 | 311.65 ± 37.97 270.60–435.54 | 2.55 | 1 | NA | 10.50 ± 0.50 6.00–12.00 | 102.13 ± 7.24 79.24–119.18 | 4.82 |
CJ20220304004 | 30 | 12.5 | 29.4 | 66.56 ± 6.15 54.40–81.60 | 299.20 ± 27.75 242.72–360.42 | 2.72 | 1 | NA | 9.00 ± 0.50 5.00–10.00 | 112.59 ± 16.92 66.89–147.06 | 4.82 |
All Individuals | 4/117 | 12.0–13.0 | 29.4–29.8 | 63.54–90.82 | 299.20–380.01 | 2.23–2.72 | 1 | NA | 9.00–11.00 | 102.13–133.77 | 4.82 |
The principal component analysis (PCA) of call parameters between L. dushanensis and L. dong resulted in two PCs, explaining a cumulated variance of 79.77% (Fig.
Factor loadings of the principal component analysis (PCA) on the advertisement call parameters of the Leptobrachella dushanensis and Leptobrachella dong.
Call parameters | Principal components | ||||
---|---|---|---|---|---|
1 | 2 | 3 | 4 | 5 | |
Call duration | 0.830 | 0.074 | -0.551 | 0.030 | 0.043 |
Inter-call interval | 0.765 | -0.395 | 0.210 | 0.463 | -0.003 |
Number of pulses | 0.967 | -0.004 | -0.178 | -0.175 | -0.060 |
Pulse rate | 0.814 | -0.192 | 0.428 | -0.341 | 0.031 |
Dominant frequency | 0.483 | 0.829 | 0.247 | 0.138 | 0.001 |
Eigenvalue | 3.104 | 0.884 | 0.624 | 0.381 | 0.006 |
Variance (%) | 62.085 | 17.686 | 12.473 | 7.628 | 0.128 |
Cumulative variance (%) | 62.085 | 79.771 | 92.244 | 99.872 | 100.000 |
From the perspective of phylogenetic relationships, L. dushanensis was assigned to the L. oshanensis species group (
This work was supported by the projects from the National Natural Science Foundation of China (Nos. 32270498, 31960099, 32260136, and 32070426), Guizhou Provincial Science and Technology Projects (Nos. ZK[2022]540 and [2023] 099), and high-level personnel research start-up funding projects of Moutai Institute (Nos. mygccrc[2022]055, mygccrc[2022]067, and mygccrc[2022]083).