Unauthorized Stocking of an Endangered Bitterling Acheilognathus typus in an Irrigation Pond Detected and Substantiated by Biological and Human Lines of Evidence
1
Aquatic Life Conservation Society, Kanazawa 12-27, Kamiyazawa, Rifu 981-0121, Miyagi, Japan
2
Fisheries Resources Institute Shiogama Branch, Japan Fisheries Research and Education Agency, Shiogama 985-0001, Miyagi, Japan
3
Biodiversity Network Niigata, Terayama 1-8-25, Higashiku, Niigata 950-0892, Niigata, Japan
4
Society for Shinaimotsugo Conservation, Koyachi 504-1, Kimazuka, Kashimadai, Osaki 989-4102, Miyagi, Japan
*
Author to whom correspondence should be addressed.
Fishes 2022, 7(4), 150; https://doi.org/10.3390/fishes7040150
Submission received: 30 May 2022
/
Revised: 18 June 2022
/
Accepted: 22 June 2022
/
Published: 25 June 2022
(This article belongs to the Section Biology and Ecology)
Abstract
:Acheilognathus typus, an endangered bitterling, was captured in an irrigation pond in the northern part of Niigata prefecture, Japan, in 2019. This bitterling species had once occupied that region. Its absence for years indicated the possible extinction of the bitterling there. We expected that the recently captured individuals are an unknown remnant stock of that endangered species found through an extensive survey. Mitochondrial genotyping, however, revealed that the recently captured individuals had a common haplotype with those from Kashimadai, Miyagi prefecture, Japan. The uniqueness of the haplotype to these two ponds indicates a stock identity between them. In the pond in Kashimadai, the illegal activity of catching the bitterlings by a person from the northern Niigata prefecture was detected in 2015. We conclude that the bitterling from the northern Niigata prefecture was stocked from a pond in Kashimadai. Our report is the first example of unauthorized bitterling stocking substantiated from both biological and human lines of evidence.
1. Introduction
Unauthorized fish stocking is defined as a clandestine activity for fish introduction without agreement or approval from stake/right holders [1]. Unfortunately, in Japan, such activities are in most cases not illegal, except for cases of legally protected/prohibited species or reserves. Unauthorized stocking is, thus, a matter of ethics mostly, though it disturbs faunal integrity in various manners, i.e., competition, predation, genetic pollution, pathogen transmission, etc. Such stocking activities involving bitterlings (Acheilognathinae, Cyprinidae) have been suspected in many water bodies in Japan [2,3,4,5,6,7]. Because bitterlings are popular in home aquaria for their beautiful coloration and small size, they often attract high prices in pet-fish markets [8,9]. Angling for bitterlings is also popular, with a particular cultural basis since the Edo period [4]. Because of the popularity of bitterlings for aquarium and angling, a strong incentive for unauthorized stocking might exist [10], especially in the case of rare and threatened species, when technically possible.
An endangered bitterling, Acheilognathus typus, is endemic to the northern part of Honshu Island (Tohoku region), Japan (Figure 1) and listed in the Japan red list as critically endangered [11]. Only a few remnant populations are currently known [12,13] after a drastic decline decades ago [14] and due to invasive largemouth bass predation [15]. It is believed extinct in the Niigata prefecture [16].
One of us (N.I.) collected this bitterling in an irrigation pond in the northern part of the Niigata prefecture in October 2019 (Figure 2 and Figure 3). The collecting site was in the Agano river–Shinano river basin that contains habitats from which this bitterling was extirpated. Hence, we genotyped the collected bitterling to test whether it was of a remnant natural population unknown to date or introduced from an existing remote habitat.
2. Materials and Methods
Five adult males (cat. no. 1901–1905) among nine collected with portable net trap and hand net were selected for mitochondrial genotyping. DNAs were extracted from fin clips with the QuickGene DNA Tissue kit and QuickGene-810 (Kurabo, Neyagawa, Japan). Primer sequences [13] and PCR/sequencing conditions [5] followed previous reports. We determined sequences of 503 bp at positions 15,640–16,142 of the complete mtDNA sequence (AB239602) [17]. This region contained the upstream 14 bp of tRNA-Pro and the upstream 489 bp of the control region.
3. Results and Remarks
Three specimens of the five (1901, 1902 and 1905) carried Hap05 (haplotype 5) that is specific to a pond in Kashimadai, Miyagi [13] (Table 1). The other two (1903 and 1904) carried Hap01 that is widely distributed over northern Honshu. The locality of the present study is approximately 160 km away from Kashimadai with a major watershed divide between them (Figure 1).
Area-endemic haplotypes of the bitterling were found among >500 specimens collected from almost all localities recently known, although overall variabilities were small [13]. Haplotype interrelationships showed a single major haplotype (Hap01) widely seen accompanying area-specific haplotypes with one nucleotide (nt) substitution from the major type except for a distant haplotype from Fukushima Aizu. The haplotype composition of the present specimens containing the Kashimadai-specific type (Hap05) was similar to the Kashimadai population (Hap01:Hap05 = 2:3 from northern Niigata vs. 17:13 from Kashimadai, p = 0.489 upon χ2-test). In addition, the T to C substitution seen in Hap05 (Table 1) was at the 2 nt linker between the upstream parts of the amino-acyl and D stems of the tRNA-Pro [18]. That site was conservative among Cyprinidae. For example, all 39 species examined had T at that site [19]. The presence of the nt substitution in the conservative site shared by two localities and its absence from other localities indicated stock identity or a close relationship between the two populations, approximately 160 km apart.
Aizu, Fukushima, the only place where a distant haplotype (>4 nt substitutions from others) was solely found [13], is an upstream basin of the Agano river running off to northern Niigata (Figure 1). Past native populations might, thus, have identical or close haplotypes with that from Aizu. If so, a similarity in the haplotype composition from the northern Niigata and Kashimadai populations might indicate the former to be non-native due to an introduction from the latter. The existence of the widely distributed haplotype, Hap01, across the watershed of a major mountain range, however, also indicated a possible fast dispersal of the bitterling within a short geological time-scale. Then, possibilities of a native distribution of the current northern Niigata population and the opposite introduction, or native distribution of both cannot be ruled out.
A suspicious case of poaching A. typus in the irrigation pond in Kashimadai was detected in 2015, where the capture and release of aquatic animals were prohibited by the city authority [20]. The suspected violator from northern Niigata confessed as a poaching trial for stocking nearby. At that time, the capture trial was stopped and the fish were safe. However, that person could learn a sense of the place to repeat the activity more easily than before, evading frequent patrol. Because people conducting unauthorized fish stocking would choose places with easy access to the stocks after establishment [21,22], the suspected violator would be highly likely to have released A. typus collected from Kashimadai into ponds in northern Niigata, near the suspected violator’s living place.
Genetic and human lines of information collectively indicated that the A. typus collected from an irrigation pond in northern Niigata in 2019 descended from the protected stock in Kashimadai. All suspected cases of the unauthorized stocking of bitterlings reported to date have had no human line of evidence [2,3,5,6,7], with one exception [4] having a human line of information only. Our report was the first example of unauthorized bitterling stocking substantiated from both biological and human lines of evidence. The presence of stocks established by unauthorized release, especially in the case of threatened fish, disturbs the prioritization of conservation targets. Without genetic information, authorities might skip the conservation of native populations, devoting limited resources instead to this introduced population.
Author Contributions
N.I. conceived this research; K.S. executed analysis and drafted the manuscript; N.I. and M.H. performed field work. All authors have read and agreed to the published version of the manuscript.
Funding
This research received no external funding.
Institutional Review Board Statement
Approved by the Research Center for Fisheries Resources, Fisheries Resources Institute, Japan Fisheries Research and Education Agency (442006, 13 June 2022). Not applicable for other institutions.
Informed Consent Statement
The person involved in the suspicious activity was informed and approved the sharing of the information upon anonymization.
Data Availability Statement
Sequences of 1901, 1902 and 1905 are identical to LC148866, and 1903 and 1904 to LC149354 can be found in the GenBank database.
Acknowledgments
We thank the personnel of the Society for Shinaimotsugo Conservation for information regarding the patrolling of the irrigation pond.
Conflicts of Interest
K.S. is a member of the editorial board of the journal and did not take any part in the review/decision process of this manuscript.
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Figure 1.
Map of the northern part of Honshu Island, Japan, showing collecting localities for Acheilignathus typus in the present (star) and previous [13] studies. The solid circle indicates a pond in Kashimadai with haplotypes identical to those from northern Niigata. Other known localities are indicated by open circles. Mountain ranges dividing watersheds are indicated by Λ. Dark gray areas contain present-day known localities, whereas areas in light gray indicate past distribution ranges.
Figure 1.
Map of the northern part of Honshu Island, Japan, showing collecting localities for Acheilignathus typus in the present (star) and previous [13] studies. The solid circle indicates a pond in Kashimadai with haplotypes identical to those from northern Niigata. Other known localities are indicated by open circles. Mountain ranges dividing watersheds are indicated by Λ. Dark gray areas contain present-day known localities, whereas areas in light gray indicate past distribution ranges.
Figure 2.
Acheilignathus typus collected from an irrigation pond in northern Niigata prefecture.
Figure 3.
An irrigation pond at the collecting site.
Table 1.
Haplotypes of the endangered bitterling Acheilognathus typus collected from an irrigation pond in northern Niigata prefecture in comparison with known types [13].
Table 1.
Haplotypes of the endangered bitterling Acheilognathus typus collected from an irrigation pond in northern Niigata prefecture in comparison with known types [13].
| Specimen/Haplotype | Variable Sites * | Geographic Endemicity | |||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| 1 | 1 | 1 | 1 | 1 | 1 | 1 | 1 | 1 | 1 | ||
| 5 | 5 | 5 | 5 | 5 | 5 | 5 | 6 | 6 | 6 | ||
| 6 | 7 | 8 | 8 | 8 | 8 | 9 | 0 | 0 | 0 | ||
| 4 | 9 | 0 | 5 | 5 | 7 | 0 | 8 | 8 | 9 | ||
| 5 | 6 | 3 | 7 | 9 | 1 | 7 | 2 | 9 | 1 | ||
| 1903 (Hap01) | T | G | T | A | T | T | G | T | T | A | |
| 1904 (Hap01) | T | G | T | A | T | T | G | T | T | A | |
| 1901 (Hap05) | C | G | T | A | T | T | G | T | T | A | |
| 1902 (Hap05) | C | G | T | A | T | T | G | T | T | A | |
| 1905 (Hap05) | C | G | T | A | T | T | G | T | T | A | |
| Known haplotypes | |||||||||||
| Hap01 | T | G | T | A | T | T | G | T | T | A | Tohoku common |
| Hap02 | T | G | T | A | T | T | G | C | T | A | Izunuma-Uchinuma |
| Hap03 | T | A | T | A | T | T | G | T | T | A | Izunuma-Uchinuma |
| Hap04 | T | G | T | A | T | T | A | T | T | A | Fukushima Hamadori |
| Hap05 | C | G | T | A | T | T | G | T | T | A | Kashimadai |
| Hap06 | T | G | T | G | T | T | G | T | T | A | Akita |
| Hap07 | T | G | T | A | T | C | G | T | T | A | Akita |
| Hap08 | T | G | C | A | C | T | G | T | G | T | Fukushima Aizu |
* Variable sites correspond with positions of A. typus complete mitochondrial sequence (AB239602) [17].
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MDPI and ACS Style
Saitoh, K.; Inoue, N.; Hasegawa, M. Unauthorized Stocking of an Endangered Bitterling Acheilognathus typus in an Irrigation Pond Detected and Substantiated by Biological and Human Lines of Evidence. Fishes 2022, 7, 150. https://doi.org/10.3390/fishes7040150
AMA Style
Saitoh K, Inoue N, Hasegawa M. Unauthorized Stocking of an Endangered Bitterling Acheilognathus typus in an Irrigation Pond Detected and Substantiated by Biological and Human Lines of Evidence. Fishes. 2022; 7(4):150. https://doi.org/10.3390/fishes7040150
Chicago/Turabian StyleSaitoh, Kenji, Nobuo Inoue, and Masatomo Hasegawa. 2022. "Unauthorized Stocking of an Endangered Bitterling Acheilognathus typus in an Irrigation Pond Detected and Substantiated by Biological and Human Lines of Evidence" Fishes 7, no. 4: 150. https://doi.org/10.3390/fishes7040150
APA StyleSaitoh, K., Inoue, N., & Hasegawa, M. (2022). Unauthorized Stocking of an Endangered Bitterling Acheilognathus typus in an Irrigation Pond Detected and Substantiated by Biological and Human Lines of Evidence. Fishes, 7(4), 150. https://doi.org/10.3390/fishes7040150
