On the Distribution and Biology of Drums (or Croakers)-sciaenidae Family-throughout the World Ocean

Zool Stud. 2019; 58: e38.

Reproductive Biology of Johnius taiwanensis (Perciformes: Sciaenidae) in Fujian Waters, Southern China

Lan-Lan Zhang

i Country Key Laboratory of Marine Environmental Science, College of Sea and World Sciences, Xiamen University, Xiamen, Fujian, Mainland china. Email: 1035451458@qq.com (Zhang); lpfang@xmu.edu.cn (Fang); 1053779442@qq.com (Xu)

Min Liu

1 State Key Laboratory of Marine Environmental Science, College of Sea and Globe Sciences, Xiamen University, Xiamen, Fujian, China. Email: 1035451458@qq.com (Zhang); lpfang@xmu.edu.cn (Fang); 1053779442@qq.com (Xu)

Lü-Ping Fang

ane State Cardinal Laboratory of Marine Environmental Science, College of Sea and World Sciences, Xiamen University, Xiamen, Fujian, Red china. Email: 1035451458@qq.com (Zhang); lpfang@xmu.edu.cn (Fang); 1053779442@qq.com (Xu)

Qing Xu

1 State Key Laboratory of Marine Environmental Science, College of Body of water and Globe Sciences, Xiamen University, Xiamen, Fujian, Communist china. East-mail: 1035451458@qq.com (Zhang); lpfang@xmu.edu.cn (Fang); 1053779442@qq.com (Xu)

Jian-Jie Lin

2 Fuzhou Marine and Fisheries Technology Center, Fuzhou, Fujian, People's republic of china. Electronic mail: 21841978@qq.com

Received 2019 Aug 26; Accepted 2019 Nov 12.

Abstract

Johnius taiwanensis is a newly described species from the Family Sciaenidae (Perciformes). The species is ordinarily found in shallow coastal waters forth both sides of the Taiwan Strait, on the west sides of Zhejiang, Fujian, Guangdong and Hong Kong and east side of Taiwan, and has been misidentified for decades. Nosotros studied the reproductive biology of J. taiwanensis from Fujian littoral waters, southern China, using gonadosomatic index (GSI) and gonad histology analyses. Monthly sampling from July 2016 to October 2017 was conducted and a total of 638 specimens were nerveless, ranging from 7.3 to 19.0 cm standard length (SL). Gonad histology suggested that the spawning activeness of J. taiwanensis females and males lasted from April to October, and the pinnacle spawning months for females was July to September. Mature females and males were 12.5 and 11.8 cm SL, respectively, while the estimated sizes at fifty% maturity were 12.0 cm and 10.9 cm SL, respectively. Vitellogenic stage oocytes (O3) and mail-ovulatory follicles (POF) or hydrated oocytes (HO) were observed, and POF and O3 in ovaries indicated that J. taiwanensis spawns multiple times each spawning flavor. HO or both HO and POF were observed in ovaries collected from one same location in May 2017 and August 2016.

Keywords: Gonad histology, Gonadosomatic index, Johnius taiwanensis, Sciaenidae, Spawning season

Groundwork

The Family Sciaenidae consists of approximately 300 species from 70 genera throughout Atlantic, Indian and Pacific Oceans, in coastal marine, brackish and freshwater; many associate with muddied or sandy bottoms (Chao et al. 2015; Lin et al. 2019). Sciaenids are important food fishes with global capture fishery production of gauge 1.7 million tons annually over the past decade (http://world wide web.fao.org/fishery/statistics/global-capture-product/en, accessed on 23 August 2019). Some sciaenids grade large aggregations for spawning, feeding and over-wintering, which makes them vulnerable to overfishing (Liu and Sadovy de Mitcheson 2008). In contempo years, small- and medium-sized croakers enter fisheries and become predominate in Chinese and adjacent waters, e.m., the genera Collichthys, Johnius and Pennahia (Yamaguchi et al. 2006; Li 2010; Tuuli et al. 2011; Wang et al. 2011; Xie et al. 2012).

The genus Johnius is endemic to Indo-West Pacific, with about 35 species, mainly occurring in shallow tropical and subtropical littoral waters, and Johnius species usually have small-scale-to medium-sized bodies and a minor rima oris, sub-concluding to inferior positioned (Trewavas 1977; Sasaki 1989 2001). Eight Johnius species are confirmed in Chinese waters, including the near recently described species, J. taiwanensis (Chao et al. 2019). Johnius taiwanensis inhabits shallow littoral waters of southeast Mainland china, along both sides of the Taiwan Strait, on the west sides of Zhejiang, Fujian, Guangdong and Hong Kong and east side of Taiwan (Chao et al. 2019) (Fig. 1). It is a mutual species simply has been misidentified for years every bit J. belangerii, J. macrorhynus, J. sina or Wak sina (Chu et al. 1963; Yu and Shen 1987; Shen 1993; Lin et al. 2007). Johnius taiwanensis can be caught by trawler and hook-and-line almost year-round and is an important local food fish.

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Distribution of Johnius taiwanensis (light purple areas) in Chinese waters. Ruby-red stars: the sampling sites from north to south, Ningde, Fuzhou, Quanzhou and Zhangzhou in Fujian waters.

As a newly described species, no studies have been conducted on the biological science or environmental of J. taiwanensis. The present study examines the reproductive cycle of J. taiwanensis in Fujian waters, southern China, using the gonadosomatic alphabetize (GSI) and gonad histology analyses, to evaluate its spawning seasonality and sizes at maturity, and to discuss the results in the context of fisheries.

MATERIALS AND METHODS

Fish sampling

Johnius taiwanensis was collected by hook-and-line in Zhangzhou waters (117°33'50"E, 23°41'xx"N), and by trawler in Quanzhou (118°57'42"E, 24°51'10"N), Fuzhou (119°46'12"E, 26°07'xvi"Due north) and Ningde (119°50'25"E, 26°36'50"Due north) waters, Fujian Province, southern China (Fig. 1). Specimens were randomly collected monthly from July 2016 to October 2017 with a continuous. Measurements of full length (TL 0.ane cm), standard length (SL 0.1 cm), whole torso weight (BW 0.1 thousand) and gonad weight (GW 0.1 g) were conducted in the laboratory.

Gonad histology

Middle portions of paired gonad lobes from males and females were fixed in Dietrich's fixative for at least one calendar week. Gonad tissue was transferred to 70% alcohol prior to histological procedures. The gonad tissue was dehydrated in a series of alcohols (from 75 to 100%), cleared in toluene, and embedded in paraffin wax. Gonad tissue were sectioned at six-7 μm, stained with haematoxylin and eosin, and mounted on slides.

Sexual maturation stages

Gonad sections were examined nether microscopy. Maturity stages of ovaries were determined based on the about avant-garde oocytes, and the occurrence of hydrated oocytes (HO), post-ovulatory follicles (POF) and/or atretic oocytes. The relative proportion of master and secondary spermatocytes (1SC/2SC) and spermatids (ST) in testes, and the appearance of sperm (SP) in sperm duct were used to determine the maturity stages of testes. Each gonad was allocated to one of the five maturity stages: young/resting female (F1) and young/resting male (M1), developing female (F2) and developing male (M2), maturing female (F3) and maturing male person (M3), ripe female person (F4) and ripe male (M4), and spent female person (F5) and spent male (M5) (Grier 1981; Wallace and Selman 1981; Yamaguchi et al. 2006; Tuuli et al. 2011) (Table 1).

Table 1

Descriptions of sexual maturation stages for females and males of Johnius taiwanensis

Sexual maturation stages Gonadal characters
Females
Immature/resting (F1) The most advanced oocytes are chief growth stage oocytes (O1). O1 closely packed and predominate.
Developing (F2) The most advanced oocytes are at cortical alveolar phase oocytes (O2), together with O1.
Maturing (F3) The most advanced oocytes are at vitellogenic phase oocytes (O3), simply prior to the nucleus migratory stage. Zona radiata is thicker than those of O1 and O2. Yolk globules begin to fuse.
Ripe (F4) The most advanced oocytes are O3 with the nucleus migratory or a unmarried yolk mass originated from the yolk globules. Hydrated oocytes (HO) or mail-ovulatory follicles (POF) may be present in some ovaries.
Spent (F5) O1 boss together with the occurrence of atretic O3 (AO3).
Males
Immature/resting (M1) Only spermatogonia (SG), primary and secondary spermatocytes (1SC/2SC) are present. No sperm in sperm duct.
Developing (M2) Big amount of 1SC/2SC with the advent of spermatids (ST). No sperm in sperm duct.
Maturing (M3) Large amount of ST at the peripheral and fundamental tubules. Sperm duct has sperm, merely not full.
Ripe (M4) Big amount of SP at the fundamental tubules. Sperm duct is full of sperm with large corporeality of ST.
Spent (M5) The lumen of tubules and sperm duct are empty or with residual sperms. SG and 1SC/2SC can be observed at the peripheral tubules.

Spawning seasonality

The spawning months were determined using both the average monthly GSI and gonad histology analyses. The GSI was calculated using the formula: GSI (%) = GW /(BW – GW) × 100. The GSI is an like shooting fish in a barrel, cheap and quick method to evaluate the spawning seasonality by identifying the peak month(due south) on GSI value. Evaluating spawning seasonality through gonad histological examination gives a more detailed description of gonad developmental stages and sexual maturation stages with a time swallow.

Size at maturity

The minimum torso sizes for female and male maturity were estimated based on GSI data and gonadal histological results. The GSI values of mature females (F3, F4 and F5) and mature males (M3, M4 and M5), adamant past gonadal histology, were plotted against their body sizes (SL); the SL with the largest and almost persistent increase in GSI% was considered to be the minimum sizes for female and male person maturity.

To determine the torso sizes at fifty% sexual maturation for females and males, all individuals collected over the spawning months were assigned to body size classes (SL) of 1.0 cm, and the percentage of mature females and males in each size class were calculated. A maturity curve was plotted against SL classes to guess the size at 50% sexual maturation.

Statistical analysis

Differences in trunk sizes of females and males were analyzed by statistical significance, set to < 0.05.

RESULTS

Biological parameters

A total of 638 individuals were collected in July 2016-October 2017; at least 30 individuals for well-nigh of the months. Body sizes ranged from seven.three to xviii.five cm SL (xiv.9 ± 1.9, hateful ± S.D., northward = 340) for females and from 8.0 to 19.0 cm SL (14.8 ± 1.nine, n = 298) for males; there was no significant departure in SL between females and males (p = 0.313) (Fig. 2). Sexual activity ratio of female: male showed monthly variation betwixt 0.69:1.00 in September and 2.50:one.00 in June, merely in that location was no significant difference (p = 0.569). The relationship of TL (cm) to SL (cm) and BW (k) to SL (cm) were: TL = 1.1366 × SL + 0.9558 (R ii = 0.9432, north = 638) (Fig. 3a) and BW = 0.02 × SL3.0198 (R ii = 0.9533, due north = 638) (Fig. 3b), respectively.

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Torso size (standard length in cm) distributions for females (due north = 340) and males (north = 298) of Johnius taiwanensis collected between July 2016 to October 2017 in Fujian waters, China.

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Relationships of (a) total length (TL in cm) to standard length (SL in cm) and (b) torso weight (BW in g) to standard length (SL in cm) of Johnius taiwanensis collected between July 2016 and October 2017 in Fujian waters, Cathay.

Sexual maturation stages

All v sexual maturation stages for females were observed: young/resting (F1), developing (F2), maturing (F3), ripe (F4) and spent (F5) (Table ane; Fig. 4). In males, developing (M2), maturing (M3), ripe (M4) and spent (M5) males were adamant; no young/resting males (M1) were constitute in the present study (Tabular array 1; Fig. five).

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Sexual maturation stages in females of Johnius taiwanensis. (a) Young/resting (F1) (17.2 cm SL, November 2016); (b) developing (F2) (15.2 cm SL, April 2017); (c) maturing (F3) (xvi.6 cm SL, September 2016); (d) ripe (F4) (16.8 cm SL, July 2016); (eastward) ripe (F4) (xvi.5 cm SL, August 2016); (f) spent (F5) (14.2 cm SL, July 2016). AO3, atretic vitellogenic phase oocyte; BV, blood vessels; GW, gonadal wall; HO, hydrated oocyte; LA, lamellae; O1, chief growth phase oocyte; O2, cortical-alveolar stage oocyte; O3, vitellogenic phase oocyte; OL, ovarian lumen; POF, mail-ovulatory follicle. Scale bars: a-c = 200 μm; d-f = fifty μm.

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Sexual maturation stages in males of Johnius taiwanensis. (a) Developing (M2) (13.1 cm SL, July 2016); (b) maturing (M3) (fourteen.4 cm SL, September 2017); (c) ripe (M4) (17.8 cm SL, September 2017); (d) spent (M5) (15.ane cm SL, Nov 2016). GW, gonadal wall; SC, spermatocytes; SD, sperm duct; SP, sperm; ST, spermatids. Scale bars = fifty μm.

Ovaries with testify of HO or both HO and POF were found in May (2017) and August (2016), nerveless in Dongshan Bay (117°33'50"East, 23°41'twenty"Northward) of Zhangzhou waters, indicating very contempo spawning activity. Ovaries had O3 and POF or O3, HO and POF, confirming that J. taiwanensis is a multiple spawner.

Spawning seasonality

The monthly boilerplate GSI% of females and males showed similar and distinct seasonal variation patterns, with females by and large having college GSI values than males (Fig. vi). In females, the boilerplate GSI peaks were in July and Baronial, with GSI over 5.v%, indicating that these were the tiptop spawning months for females. From November to April, the average GSI% of females were depression, less than 1.5%. In males, the average GSI peaks were in May and August with GSI over 1.3%, indicating that this was the peak spawning season for males. Similar to females, the average GSI% of males were low from November to adjacent April, less than 0.6%.

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Monthly gonadosomatic index (GSI%, hateful ± S.D.) in females and males of Johnius taiwanensis collected between July 2016 and October 2017.

Based on gonadal histology, the spawning seasons for females and males were revealed to concluding longer than those determined by GSI values above. In females, F1 and F2 were establish from November to June, dominating from November to next Apr (Fig. 4a b; Fig. 7a). The proportion of F2 increased in March and April, followed by the appearance of F3 in April and the increase of F3 in May-August, indicating that this was the ovary development phase of female sexual maturation (Fig. 4c; Fig. 7a). The proportions of F3 and F4 remained high between May and October, indicating the female spawning season in May−October. In July−September, the proportions of F4 were more than than 50%, indicating the peak spawning season (Fig. 4d e; Fig. 7a). The proportion of F5 and F1 were high after October, indicating the terminate of the spawning season (Fig. 4f; Fig. 7a).

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Monthly female (a) and male (b) sexual maturation stages of Johnius taiwanensis based on gonad histology. F1, immature/resting female; F2, developing female person; F3, maturing female; F4, ripe female person; F5, spent female; M2, developing male; M3, maturing male; M4, ripe male; M5, spent male person. The criteria for sexual maturation stages were described in tables ane and 2. Numbers on the tiptop of confined represent sample sizes.

M1 was not found in males. M2 was simply establish in July (Fig. 5a; Fig. 7b). M3 was absolutely dominant in March (100%), and decreased with the dramatically increase of M4 proportions in Apr and May (Fig. 5b c; Fig. 7b). M4 remained loftier between Apr and October, indicating that the male spawning season was April−October, longer than that of females. Loftier proportions of M4 (> 50%) in April-September indicated the peak spawning season. M5 were ascendant from October to side by side February, indicating the finish of the spawning season (Fig. 5d; Fig. 7b).

Size at maturity

The estimated minimum body sizes for female and male person maturity were 12.5 and 11.8 cm SL, respectively (Fig. 8a b). The estimated body sizes at 50% sexual maturation for females and males were 12.0 and 10.nine cm SL, respectively, based on all females and males collected during the spawning months from Apr to October (Fig. 9).

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Relationship of the gonadosomatic index (GSI%) and standard length (SL) for (a) mature females (north = 204) and (b) mature males (n = 260) of Johnius taiwanensis. F3, maturing female person; F4, ripe female person; F5, spent female person; M3, maturing male; M4, ripe male; M5, spent male person. Vertical lines represent the minimum sizes with big increment in GSI%: 12.5 cm SL in (a) and 11.8 cm SL in (b).

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Sizes at 50% sexual maturation for females and males of Johnius taiwanensis, based on all females (due north = 204) and males (n = 198) nerveless during the spawning months from April to October. 12.0 cm and ten.ix cm SL were extrapolated straight for females and males, respectively.

Word

In Chinese waters, 27 sciaenid species are confirmed from xiv genera (Argyrosomus, Atrobucca, Bahaba, Chrysochir, Collichthys, Dendrophysa, Johnius, Larimichthys, Megalonibea, Miichthys, Nibea, Otolithes, Pennahia, Sciaenops), including S. ocellatus, which was introduced from the The states for aquaculture purposes in the 1990s and has invaded Chinese waters (Liao et al. 2009; Chao et al. 2019). Most of these sciaenid species are pocket-size- and medium-sized, with only domestic and local commercial importance. The maximum size of J. taiwanensis in the present study was a male person of 19.0 cm SL, collected in Zhangzhou waters (Fig. ane; Fig. 2). Information technology is confirmed to be the largest individual recorded to date because the maximum size among holotype and paratype specimens was xviii.6 cm SL (Chao et al. 2019).

In Chinese and adjacent waters, the spawning seasonality of sciaenids has been examined in a few species based on either gonad external morphology merely, or GSI% and/or gonad histology analyses (Chu 1960; Chu and Wu 1985; Kakuda and Nakai 1981; Li et al. 2000; Li 2010; Lin et al. 2008; Ni 2018; Tuuli et al. 2011; Wang et al. 2011 2012; Wu 1981; Xu 2014; Yamaguchi et al. 2006). The results showed that the spawning activity of nigh species examined lasted iii−8 months, e.k., Chrysochir, Collichthys, Dendrophysa, Johnius, Larimichthys, Nibea, Otolithes and Pennahia species (Table 2). To the north, the spawning flavour in the E China Sea was mainly dominant in Leap (April and May), due east.g., for 50. crocea and L. polyactis, while to the south in the Taiwan Strait and northern South China Sea, the spawning seasons lasted longer, from Bound to Autumn, mainly dominant in May−October, e.thousand., for C. aureus, C. lucidus, D. russelii, J. belangerii, J. taiwanensis, L. crocea and P. macrocephalus. In the present study, the spawning menstruation of J. taiwanensis lasted from April to October in Fujian waters, Mainland china, with the top in July−September for females, based on gonadal histology analyses.

Tabular array ii

Spawning seasons of sciaenid species in Chinese and next waters

Species Spawning flavour (acme season) Methodology Area Reference
Chrysochir aureus June to December (July to September) GSI and gonadal histology Southwestern waters of Taiwan Ni 2018
Collichthys lucidus Jan and Apr Gonad external morphology Fujian waters, Communist china Wang et al. 2011
March to December Gonad external morphology Pear River Estuary of Guangdong, China Li et al. 2000
Dendrophysa russelii March to September (March to May) GSI and gonadal histology Liusha Bay of Guangdong, People's republic of china Xu 2014
Johnius belangerii June to September (July and Baronial) Gonad external morphology Zhoushan waters of Zhejiang, China Wang et al. 2012
Twelvemonth-round (April and August) Gonad external morphology Fujian waters, Cathay Wang et al. 2011
May to July Gonad external morphology Pear River Estuary of Guangdong, China Li et al. 2000
March to Oct (March) GSI and gonadal histology Liusha Bay of Guangdong, China Xu 2014
J. taiwanensis April to October (July to September) GSI and gonadal histology Fujian waters, China The present study
Larimichthys crocea October to Dec Gonad external morphology Hong Kong waters Chu 1960
October to December Gonad external morphology Liusha Bay of Guangdong, China Li et al. 2000
April to June - E Cathay Body of water Chu and Wu 1985
L. polyactis Apr to June GSI and gonad histology Bohai and the northern xanthous Sea Wu 1981
April and May Gonad external morphology Southern Yellow Sea Lin et al. 2008
March to May Gonad external morphology Eastward Cathay Sea Lin et al. 2008
Nibea albiflora May to August (June and July) GSI and gonadal histology Seto Inland Sea, Japan Kakuda and Nakai 1981
Otolithes ruber April to July (April to July) GSI and gonadal histology Southwestern waters of Taiwan Ni 2018
Pennahia anea March to June (May) GSI and gonadal histology Hong Kong waters, Mainland china Tuuli et al. 2011
P. argentata April to September (May to August) GSI and gonadal histology Ariake Audio waters, Nihon Yamaguchi et al. 2006
P. macrocephalus May to October (July to September) GSI and gonadal histology Southwestern waters of Taiwan Li 2010
April and Baronial Gonad external morphology Fujian waters, Cathay Wang et al. 2011

-: no data.

The minimum sizes at maturity or the sizes at 50% maturity for females and males are the two criteria commonly used for evaluating the maturity sizes of the fish. For most studies on sciaenid species, mature females are generally larger than mature males. For example, the estimated minimum sizes of mature females and males of P. anea are 12.five cm and eleven.nine cm SL, respectively, and the size at 50% maturity for females was 12.3 cm SL (Tuuli et al. 2011). For P. macrocephalus, the sizes at 50% maturity for females and males are fifteen.9 cm and 14.eight cm SL, respectively (Li 2010). For C. aureus, the sizes at 50% maturity for females and males are 26.3 cm and 21.0 cm SL, respectively (Ni 2018). For O. ruber, the sizes at fifty% maturity for females and males were 21.8 cm and 19.9 cm SL, respectively (Ni 2018). In a few species, mature females were smaller than mature males. For instance, in J. belangerii, the sizes at 50% maturity for females and males were 12.five cm and thirteen.six cm SL, respectively (Wang et al. 2012). In the present study, the estimated minimum sizes for female and male maturity of J. taiwanensis were 12.5 cm and 11.8 cm SL, respectively, mature females larger than mature males. The estimated torso sizes at l% sexual maturation for females and males of J. taiwanensis were 12.0 cm and 10.9 cm SL, respectively, smaller than the minimum sizes for female person and male maturity.

Johnius taiwanensis is a multiple spawners in a spawning season based on gonadal histology, just similar other sciaenid species reported. The show includes the appearance of dissimilar developmental stages of oocytes from primary growth to vitellogenic in ovaries and sometimes beyond vitellogenic such equally HO and POF (Kakuda and Nakai 1981; Li 2010; Ni 2018; Tuuli et al. 2011; Wu 1981; Xu 2014; Yamaguchi et al. 2006; the present study).

Spawning grounds of some sciaenid species in Chinese waters were associated with estuaries. For B. taipingensis, the spawning grounds were associated with the Yangtze River, the Min River and the Pearl River Estuaries (Sadovy and Cheung 2003). For L. crocea, the spawning grounds were also associated with the river mouths along the Chinese declension (Liu and Sadovy de Mitcheson 2008). The advent of HO or both HO and POF in ovaries of J. taiwanensis was confirmed based on gonadal histology, indicating the samples were from its spawning basis. The spawning ground was subsequently confirmed in Dongshan Bay (117°33'50"E, 23°41'20"N) of Zhangzhou waters, associated with the Zhangjiang River Estuary.

CONCLUSIONS

Johnius taiwanensis is one of the few remaining wild-caught croakers commonly plant in the Taiwan Strait. In the present study, the spawning seasonality, the minimum sizes for female person and male maturity and the sizes at l% maturity of J. taiwanensis were determined. The spawning season of J. taiwanensis lasted from April to Oct, peaking in July-September for females. Currently, the nationwide management for domestic marine fisheries has a fishing moratorium in May-August, adjusting slightly for fishing gears. Therefore, J. taiwanensis can exist partly protected under the current management measures. Future studies should further investigate the locations of J. taiwanensis spawning grounds.

Acknowledgments

The present study was supported past the National Programme on Global Modify and Air-Body of water Interaction (GASI-02-PAC-YDaut), and Cardinal Research Funds for Central Universities from Xiamen University (grant no. 20720170077). The authors would like to thank Nian-Ping Cai, Wei-Di Yang, Ying Su, Yuyan Jia, Baian Lin, Xiao-Bing Jiang and Yan Jiang for field and laboratory back up. We also thank Prof. Ning Labbish Chao for specimen identification and Dr. Claire Gorman for providing the distribution map.

Footnotes

Authors' contributions: LLZ and ML designed the report. LLZ wrote the outset draft of the manuscript and performed information analyses. LLZ, LPF and JJL conducted the field collection. LLZ and QX conducted laboratory measurement, dissection and gonadal histology. ML and LPF revised the manuscript. All authors read and approved the last manuscript.

Competing interests: The authors declare that they have no competing interests.

Availability of data and materials: All the data and materials are provided inside the manuscript.

Consent for publication: Not applicable.

Ideals approval consent to participate: Not applicable.

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Source: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6971531/

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