Indian Journal of Animal Research

  • Chief EditorK.M.L. Pathak

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DNA Barcoding of Argyrops filamentosus (Perciformes: Sparidae) Based on Mitochondrial COI Gene and Histopathological Evaluation of Gills Infected by Monogeneans

Rewaida Abdel-Gaber1,*, Saleh Al Quraishy1, Masheil Alghamdi1, Hossam M.A. Aljawdah1, Saleh N. Maodaa1, Esam M. Al-Shaebi1
1Department of Zoology, College of Sciences, King Saud University, Riyadh, Saudi Arabia.

Background: Sparidae is a fish family that belongs to the order Perciformes and is commonly known as sea breams and porgies. Many species are included in this family and divided into 71 genera that could be barcoded using mitochondrial DNA genes. These fish are vulnerable to different parasitic taxa that affect fish status. The present study is aimed to confirm the molecular status of Argyrops filamentosus fish via the mitochondrial cytochrome c oxidase (COI) gene and to study the pathological changes of gills infested by monogenean parasites.

Methods: Thirty Argyrops filamentosus fish were collected from Jeddah (Saudi Arabia) and identified molecularly via the mtCOI gene. Also, gills were isolated and examined microscopically for presence of monogeneans. Histopathological impacts of monogeneans on fish gills were studied in comparison to the gills of non-infected fish. 

Result: The DNA of fish species was barcoded and showed highly stringent criteria with the previously Argyrops filamentosus sequence data. The obtained host DNA sequences were deposited in NCBI database under accession number OP975758.1. Examination of the investigated fish gills revealed the presence of three monogenean species Protolamellodiscus senilobatus Kritsky, Jiménez-Ruiz and Sey, 2000, Acleotrema maculatus Morsy, El-Fayoumi and Fahmy (2014) and Haliotrema susanae Soo, 2019. Monogenean parasites penetrated deeply with their haptor to the gill lamella and caused damage and degeneration of epithelial cells leading to the formation of a cup-shaped depression. Therefore, the mtDNA gene has the ability for host identification and heavy monogeneans infections lead to severe damage to fish gills. 

The king soldier bream Argyrops filamentosus Valenciennes, 1830, belongs to the family Sparidae (Order: Perciformes) and this species is originally distributed in the western Indian Ocean from the Red Sea, Oman and Arabian Gulf to South Africa including Mauritius, Madagascar and La Réunion (Fricke et al., 2018). The soldier bream is an active carnivorous species feeding particularly on sardines and benthic invertebrates (Ghanem et al., 2021). Fish are well known to be parasitized by many eukaryotic organisms (M’Rabet et al., 2016) and the gills of fish represent one of the biotopes mostly exploited by different parasites, including monogeneans (Shawket et al., 2018).

Traditional morphology-based species identification is being replaced by DNA barcoding because it is becoming more generally available, reliable and is more widely adopted (Ali et al., 2014). According to Nerlovic et al., (2015), the partial examination of the mitochondrial cytochrome c oxidase subunit I (COI) and 16S ribosomal RNA (16S rRNA) gene sequences validated the fish’s identity. DNA-based techniques were used by Hendrick et al., (2019) and Cucalon et al., (2022) to guarantee successful host identification.

The knowledge of parasitic communities makes researchers recognize the role of host fish in the food web or ecosystem and provide a deeper assessment of the role of parasites in affecting fish health (Shawket et al., 2018). The gills in fish are important organs of examination in disease diagnosis, because of their direct contact with the environment, such as several irritants, parasites and pollutants present in the water. The host and parasite relationship in the case of monogeneans results in large-scale damage on the site of attachment (Shawket et al., 2018). To attach themselves to a host, monogeneans use a specialized organ called a haptor (Arya and Singh, 2020). By attachment and feeding, monogeneans induce pathological changes in the host epithelium, thus creating a gateway for secondary bacterial and viral infections (Feist and Longshaw, 2008).

The present study is aimed to confirm the molecular status of A. filamentosus fish targeting the COI gene and to study the pathological changes of gills infected by monogenean parasites in comparison with non-infected gills.
Fish and samples collection
 
Thirty fish specimens of the solider bream, A. filamentosus were collected from commercial fishermen in Jeddah (Saudi Arabia), during the period January-December 2022. The fish’s gills were separated, submerged in 0.9% saline solution, then examined under Nikon SMZ18 stereomi croscope. Monogeneans were isolated from the gills and identified according to the diagnostic criteria. A piece of fish muscle was removed from the dorsal side and preserved in 96% ethanol for molecular analysis.
 
Molecular barcoding of fish
 
DNA was extracted using a DNeasy tissue kit© (Qiagen, Hilden, Germany) following the recommended steps. From the mitochondrial genome, the cytochrome c oxidase I (COI) gene was amplified by PCR using primers of Baldwin et al., (2009). PCR amplification under the following conditions: 95°C for 3 min, followed by 35 cycles at 95°C for 20 s, 55°C for 20 s and 72°C for 40 s, with a final extension at 72°C for 10 min. PCR products were sequenced using ABI PRISM 310 Genetic Analyzer (Applied Biosystems, Foster City, CA) at Prince Naif Health Research Center, King Saud University, Riyadh, Saudi Arabia. Sequences were deposited at a public sequence database, GenBank of the National Center for Biotechnology Information (NCBI, https://www.ncbi.nlm.nih.gov/). A search on BLASTn was performed to determine the most related sequences available in GenBank for the COI dataset. Analysis of the aligned DNA sequences of the partial COI gene region was done by the maximum likelihood (ML) using MEGA7 with the best-fit substitution models. Statistical support for each node was evaluated using a non-parametric bootstrap test with 1000 replicates.
 
Histopathological studies
 
Small portions of gills from non-infected fish and those naturally infected with monogeneans were collected and fixed in 10% neutral buffered formalin. The tissues were processed by routine paraffin embedding method and the sections were stained with Hematoxylin and Eosin (H & E). The stained sections were examined and photographed using a Leica DM 2500 microscope (NIS ELEMENTS software, version 3.8).
 
Prevalence of parasitic infections
 
Fish are considered one of the important sources of animal protein (de Boer et al., 2020). Sea breams are fish of considerable economic importance and are commonly affected by several pathological problems, which can lead to mortalities and loss of production (Bannai, 2018). The examined solider bream fish, A. filamentosus, was found to be naturally infected with three Monogeneans, ectoparasitic flatworms. Identification of parasitic taxa of sea breams associated with their morphological and biological peculiarities. Molecular techniques provide useful alternatives to morphology for the identification of parasites (Chaudhary et al., 2015; Abdel-Gaber et al.,  2023). Monogeneans isolated from the gills of the solider bream fish were identified as Protolamellodiscus senilobatus Kritsky, Jiménez-Ruiz and Sey, 2000 (F: Diplectanidae), Acleotrema maculatus Morsy, El-Fayoumi and Fahmy (2014) (F: Diplectanidae) and Haliotrema susanae Soo, 2019 (F: Ancyrocephalidae) with a prevalence rate of 70 (21/30) for P. senilobatus, 46.66% (14/30) for A. maculatus and 50.00% (15/30) for H. susanae. Mixed infection for the recovered parasites was recorded to be 33.33% (10/30). Little information is available on parasitic infections of the soldier bream fish and the specificity with its host. Therefore, the present study provided more data on parasite fauna in this sparid fish species and confirm its taxonomic position.  
 
Molecular barcoding of Argyrops filamentosus fish
 
DNA barcoding is considered to be an efficient tool to identify fish species within an ecosystem and to evaluate the genetic variability within species (Rajkumar et al., 2015; Panprommin et al., 2019; Ghouri et al., 2020; Alshehri et al., 2022). In the present study, the partial sequences of the mt COI gene were amplified by PCR that generated 539 bp fragment. The average base-pair proportions for the COI gene were A(24.49% 132) | C(28.01% 151) | G(16.14% 87) | T(31.35% 169), which is consistent with Ward et al., (2005), Ivanova et al., (2007), Armani et al., (2012), Consuegra et al., (2015) and Cutarelli et al., (2018). PCR products were deposited in GenBank under the accession number OP975758.1. The ML approach was used to align nucleotide sequence data from 24 taxa over 530 positions to produce a phylogenetic dendrogram that represented different marine species (Table 1).

Table 1: GenBank accession numbers for COI sequences used in ML analysis for fish host.



There is a low intra-species variation (0.076) for COI among all specimen sequences, agreeing with Xia et al., (2008), Renxie et al., (2018) and Ceruso et al., (2020) who also observed the lower intraspecific distance between comparable sparid fish. The results validate that COI is a useful marker for species diagnosis in the family Sparidae. One family (Sparidae) for the fish host (Argyrops filamentosus) was identified for the recovered monogenean parasites. The phylogenetic relationship of the family Sparidae is supported by previous data from Day (2002) and Chiba et al., (2009). All the GenBank entries that matched our COI sequence under the highly stringent criteria (89.90-99.44% identity, 98-100% query coverage and E-value 0.0) were assigned to two monophyletic groups within family Sparidae for A. spinifer and A. filamentosus. In ML analyses (Fig 1), the taxa of Argyrops were grouped in a distinct clade with high bootstrap values. Dendrograms confirmed the association of our specimen with the Argyrops group, with special reference to Argyrops filamentosus (JF492876.1, KJ012288.1, KJ012287.1, HQ945946.1 and HQ945852.1) that found in South Africa and the Indian Ocean. This study confirms the high efficacy of DNA barcodes for the identification of sparid fish.

Fig 1: Phylogenetic analysis by Maximum Likelihood method based on the Tamura-Nei model.


 
Histopathological studies
 
The histopathological alterations of the infected gills were observed and compared to the control sections of A. filamentosus (Fig 2).

Fig 2: Gills isolated from Argyrops filamentosus.



The damages are consistent with Purivirojkul (2012) reported that infection with monogeneans is one of the most prevalent parasitic agents affecting gills and destroying gills which leads to impairment of fish respiration as well as a general deterioration of fish health. Microscopic examinations revealed that the gills from control fish consisted of horizontal flat filaments and were supported by bony gill arches. On these filaments, secondary lamellae were found. These lamellae are composed of a thin epithelial cell layer covering pillar cells, which in turn surround the blood sinusoids. Monogenean parasites were attached to the secondary gill lamella of infected fish. Parasites penetrated deeply with their haptor to the gill lamella and caused damage and degeneration of epithelial cells leading to the formation of a cup-shaped depression. Nevertheless, the major host response to monogenean infection was represented by hyperplasia of the gill filament. The severity of respiratory damage for the fish host was directly proportional to the number of parasites on the gills. This agreed with Campos et al., (2011) and Pahor et al., (2017) stated that when the intensity of monogenean parasites increases, the gill damage may be serious with the pronounced impact of the histology and lead to fish mortality. Because of these damages, hyperplasia of gill epithelium associated with monogenean anchors was observed, which agreed with Shamsi et al., (2009), Raissy et al., (2011), Pahor et al., (2017) and Hoai (2020) also noticed hyperplasia of the epithelial cells, subsequent lamellar fusion and extensive secretion of mucus. Moreover, these results agreed with those observed on the gill filament of Plectropomus maculatus infecting Acleotrema maculatus (Morsy et al., 2014), Pagellus erythrinus infecting Diplectanum aequans (Adawy et al., 2016), Gyrodactylus cichlidarum infecting Oreochromis niloticus (Grano-Maldonado et al.,  2018), Cichlidogyrus philander infecting Pseudocrenilabrus philander (Igeh and Avenant-Oldewage, 2020), Dactylogyrus spp. infecting Epinephelus coioides (Ramudu et al., 2020); and three species of monogeneans Anacanthorus spathulatus, Notozothecium janauachensis and Mymarothecium boegeri infecting Colossoma macropomum (Tavares-Dias et al., 2021).  
Therefore, it could be concluded that the ability of mtDNA barcoding for host identification determines the specificity of monogenean parasites to their host species. There is an effect of heavy parasitic infections on fish gills and epithelial destruction of lamellae. Further studies should include some tools for controlling the parasite distribution.
This study was supported by the Researchers Supporting Project (RSP2023R25), King Saud University, Riyadh, Saudi Arabia.
All the datasets generated or analyzed during this study are included in this published article.
The author(s) declare that they have no conflict of interest regarding the content of this article.

  1. Abdel-Gaber, R., Alotaibi, M., Al Quraishy, S., Omar, S.A., Elkhadragy,  M.F., Aljawdah, H.M.A., Al-Shaebi, E.M. (2023). Monosteph anostomum mesospinosum (Digenea: Acanthocolpidae) infecting the coastal travelly Carangoides caeruleopinnatus (Carangiformes: Carangidae) from the marine water off Saudi Arabia. Indian Journal of Animal Research. pp. 1-9.

  2. Adawy, R., Abd El-Meged, R., Sorour, S.S.G. (2016). Some studies on monogenean infections in gills of marine water fishes in Egypt. Assiut Veterinary Medical Journal. 62(151): 1-11. 

  3. Ali, A.M., Gyulai, G., Hidvégi, N., Kerti, B., Al Hemaid, F.M., Pandey, A.K., Lee, J. (2014). The changing epitome of species identification - DNA barcoding. Saudi J. Biol Sci. 21(3): 204-231. 

  4. Alshehri, E., Alqahtani, J., Al-Shaebi, E.M., Al-Quraishy, S., Abdel- Gaber, R. (2022). Identification of Gnathiid isopods as ectoparasites of the Brown-spotted Grouper Epinephelus chlorostigma from thr Red Sea of Saudi Arabia. Indian Journal of Animal Research. pp.1-8. 

  5. Armani, A., Guardone, L., Castigliego, L., D’Amico, P., Messina, A., Malandra, R., Gianfaldoni, D., Guidi, A. (2012). DNA and Mini-DNA barcoding for the identification of Porgies species (family Sparidae) of commercial interest on the international market. Food Control. 50: 589-596. 

  6. Arya, P.V., Singh, H.S. (2020). Microhabitat preference studies of freshwater fish gill parasitic monogenea Mizelleus indicus (Jain, 1957), Pandey et al. (2003). on gills of Wallago attu (BI. And Sch.). Biochemical and Cellular Archives. 20: 3945.  

  7. Baldwin, C.C., Mounts, J.H., Smith, D.G., Weigt, L.A. (2009). Genetic identification and color descriptions of early life- history stages of Belizean Phaeoptyx and Astrapogon (Teleostei: Apogonidae) with comments on identification of adult Phaeoptyx. Zootaxa. 2008: 1-22.

  8. Bannai, M.A.A. (2018). Hysterothylacium persicum (Nematoda: Raphidascarididae) parasite of orangespotted grouper Epinephelus coioides (Forsskål, 1775) Iraqi marine water fishes. Iraqi Journal of Science. 59(3C): 1548-1533.

  9. Campos, C.M., Moraes, J.R.E., Moraes, F.R. (2011). Histopathology of gills of Piaractus mesopotamicus (Holmberg, 1887) and Prochilodus lineatus (Valenciennes, 1836) infested by monogenean and myxosporea, caugth in Aquidauana River, State of Mato Grosso do Sul, Brazil. Revista Brasileira de Parasitologia Veterinária0 20(1): 67-70. 

  10. Ceruso, M., Venuti, I., Osca, D., Caputi, L., Anastasio, A., Crocetta, F., Sordino, P., Pepe, T. (2020). The complete mitochondrial  genome of the sharpsnout seabream Diplodus puntazzo (Perciformes: Sparidae). Mitochondrial DNA B Resour. 5(3): 2379-2381.

  11. Chaudhary, A., Chiary, H.R., Sharma, B., Singh, H.S. (2015). First report on molecular identifi cation of invasive tapeworm, Bothriocephalus acheilognathi Yamaguti, 1934 (Cestoda: Pseudophyllidea) in India. Bioinvasions Rec. 4: 269-276.

  12. Chiba, S., Iwatsuki, Y., Yoshino, T., Hanzawa, N. (2009). Comprehensive  phylogeny of the family Sparidae (Perciformes: Teleostei) inferred from mitochondrial gene analyses. Genes and Genetic Systems. 84(2): 153-170. 

  13. Consuegra, S., John, E., Verspoor, E., de Leaniz, C.G. (2015). Patterns of natural selection acting on the mitochondrial genome of a locally adapted fish species. Genet Sel Evol. 47: 58.(2015). https://doi.org/10.1186/s12711-015- 0138-0. 

  14. Cucalon, R.V., Valdiviezo-Rivera, J., JiménezPrado, P., Navarrete- Amaya, R., Shervette, V.R., Torres-Noboa, A., Wierzal, N., Karpan, K.C., Borders, T., Calle, P., Lujan, N.K., Aguirre, W.E. (2022). Phylogeography of the Chocó endemic rainbow characin (Teleostei: Rhoadsia). Ichthyol Herpetol. 110(1): 138-155.

  15. Cutarelli, A., Galiero, G., Capuano, F., Corrado, F. (2018). Species identification by means of mitochondrial cytochrome b DNA sequencing in processed Anchovy, Sardine and Tuna species. Food and Nutrition Sciences. 9: 369-375. 

  16. Day, J.J. (2002). Phylogenetic relationships of the Sparidae (Teleostei:  Percoidei) and implications for convergent trophic evolution.  Biol J. Linn Soc. 76: 269-301.

  17. de Boer, J., Schösler, H., Aiking, H. (2020). Fish as an alternative protein - A consumer - oriented perspective on its role in a transition towards more healthy and sustainable diets. Appetite. 152: 104721. 

  18. Feist, S.W., Longshaw, M. (2008). Histopathology of fish parasite infections - importance for populations. Journal of Fish Biology. 73: 2143-2160.

  19. Fricke, R., Mahafina, J., Behivoke, F., Jaonalison, H., Léopold, M., Ponton, D. (2018). Annotated checklist of the fishes of Madagascar, southwestern Indian Ocean, with 158 new records. Fish Taxa. 3(1): 1-432. 

  20. Ghanem, R., Jamila, B.S.S., Azzurro, E. (2021). Documented occurrence of the exotic soldierbream Argyrops filamentosus  (Valenciennes, 1830) in the specially protected area of Zembra (Tunisia): a result of a joint monitoring strategy. Cahiers de Biologie Marine. 62: 227-233. 

  21. Ghouri, M.Z., Ismail, M., Javed, M.A., Khan, S.H., Munawar, N., Umar, A.B., Nisa, M.U., Afrab, S.O., Amin, S., Khan, Z., Ahmad, A. (2020). Identification of edible fish species of Pakistan through DNA barcoding. Front Mar Sci. 7: 554183. 

  22. Grano-Maldonado, M.I., Rodríquez-Santiago, M.A., García-Vargas, F., Nieves-Soto, M., Soares, F. (2018). An emerging infection caused by Gyrodactylus cichlidarum Paperna, 1968 (Monogenea: Gyrodactylidae) associated with massive mortality on farmed tilapia Oreochromis niloticus (L.) on the Mexican Pacific coast. Lat Am J. Aquat Res. 46(5): 961-968. 

  23. Hendrick, G.C., Dolan, M.C., McKay, T., Sikkel, P.C. (2019). Host DNA integrity within blood meals of hematophagous larval gnathiid isopods (Crustacea, Isopoda, Gnathiidae). Parasites and Vectors. 12: 316. 

  24. Hoai, T.D. (2020). Reproductive strategies of parasitic flatworms (Platyhelminthes, Monogenea): the impact on parasite management in aquaculture. Aquaculture International. 28: 421-447.  

  25. Ivanova, N.V., Zemlak, T.S., Hanner, R.H., Hebert, P.D.N. (2007). Universal primer cocktails for fish DNA barcoding. Molecular Ecology Notes. 7(4): 544-548. 

  26. Kritsky, D.C., Jiménez-Ruiz, F.A., Sey, O. (2000). Diplectanids (Monogenoidea: Dactylogyridea) from the gills of marine fishes of the Persian Gulf off Kuwait. Comparative Parasitology. 67(2): 145-164. 

  27. M’Rabet, C., Ensibi, C., Dhaouadi, R., Yehia, O.K. (2016). A preliminary study on gill parasites of gilthead sea bream Sparus aurata (Linnaeus 1758) (Pisces: Teleostei) from the eastern Tunisian sea-cage aquaculture. GERF Bull Biosci. 7(1): 1-5. 

  28. Morsy, K., El Fayoumi, H., Fahmy, M. (2014). Description of Acleotrema maculatus sp. nov. (Monogenea: Diplectanidae)  infecting the spotted coral grouper Plectropomus maculatus (F: Serranidea) from the Red Sea and its histopathological impact. Beni-Suef University Journal of Basic and Applied Sciences. 3(3): 222-228. 

  29. Nerlovic, V., Mravinac, B., Devescovi, M. (2015). Additional information  on the blue runner, Caranx crysos (Mitchill, 1815), from the northern Adriatic Sea: meristic and molecular characterizations. Acta Adriat. 56: 309-318.

  30. Pahor, E.F., Klosterhoff, M.C., Marchiori, N.C., Pereira, J. (2017). Moderate pathogenic effect of Ligophorus uruguayense (Monogenoidea, Ancyrocephalidae) in juvenile mullet Mugil liza (Actinopterygii, Mugilidae) from Brazil. Anais da Academia Brasileira de Ciências. 89(4): 2997-3003.

  31. Panprommin, D., Soontornprasit, K., Tuncharoen, S. (2019). DNA barcodes for the identification of species diversity in fish from Kwan Phayao, Thailand. Journal of Asia-Pacific Biodiversity. 12(3): 382-389.

  32. Purivirojkul, W. (2012). Histological change of aquatic animals by parasitic infection, histopathology-Reviews and Recent Advances, Enrique Poblet Martinez, Intech Open. https:/ /doi.org/10.5772/52769. https://www. intechopen.com/ books/histopathology-reviews-and-recent-advances/ histological-change-of-aquatic-animals-by-parasitic- infection.

  33. Raissy, M., Ansari, M., Moumeni, M. (2011). Parasite fauna of Aphanius vladykovi Coad, 1988 (Osteichthyes: Cyprinodontidae) in Gandoman Lagoon. Comp Parasitol. 78: 104-106.

  34. Rajkumar, G., Bhavan, P.S., Udayasuriyan, R., Vadivalagan, C. (2015). Molecular identification of shrimp species, Penaeus  semisulcatus, Metapenaeus dobsoni, Metapenaeus brevicornis, Fenneropenaeus indicus, Parapenaeopsis stylifera and Solenocera crassicornis inhabiting in the coromandel coast (Tamil Nadu, India) Using MT-COI gene. International Journal of Fisheries and Aquatic Studies. 2(4): 96-106. 

  35. Ramudu, K.R., Shivam, S., Sanil, N.K., Sharma, S.R.K., Vijayagopal,  P., Babu, P.P.S., Anuraj, A., Navanath, K., Rao, B.M., Loka, J. (2020). Histopathological changes in the gills associated with Dactylogyrus spp. infestation in Orange- spotted grouper, Epinephelus coioides reared in Sea Cages. International Journal of Current Microbiology and Applied Sciences. 9(8): 1-8. 

  36. Renxie, W., Haoran, Z., Jing, L., Sufang, N., Yao, X., Yongxia C. (2018). DNA barcoding of the family Sparidae along the coast of China and revelation of potential cryptic diversity in the IndoWest Pacific oceans based on COI and 16S r- RNA genes. J. Oceanol Limn. 36: 1753-1770.

  37. Shamsi, S., Jalali, B., Aghazadeh Meshgi, M. (2009). Infection with Dactylogyrus spp. among introduced cyprinid fishes and their geographical distribution in Iran. Int J. Vet Res. 10: 70-74.

  38. Shawket, N., Elmadhi, Y., M’bareck, I., Youssir, S., El Kharrim, K. and Belghyti, D. (2018). Distribution of two monogenean (Gastrocotylidae) from the North Atlantic coast of Morocco. Beni-Suef University Journal of Basic and Applied Sciences.  7(3): 270-275.

  39. Soo, O.Y.M. (2019). A new species of Haliotrema (Monogenea: Ancyrocephalidae (sensu lato) Bychowsky and Nagibina, 1968) from holocentrids off Langkawi Island, Malaysia with notes on the phylogeny of related Haliotrema species. Parasitol Int. 68(1): 31-39.

  40. Tavares-Dias, M., Ferreira, G.V., Videira, M.N. (2021). Histopathological  alterations caused by monogenean parasites the gills of tambaqui Colossoma macropomum (Serrasalmidae). Semina: Ciênc Agrár. 42(3): 2057-2064. 

  41. Ward, R.D., Zemlak, T.S., Innes, B.H., Last, P.R., Hebert, P.D. (2005). DNA barcoding Australia’s fish species. Philos Trans R Soc Lond B Biol Sci. 360(1462): 1847-1857. 

  42. Xia, J.H., Huang, J.H., Gong, J.B., Jiang, S.G. (2008). Significant population genetic structure of yellowfin seabream Acanthopagrus latus in China. Journal of Fish Biology. 73(8): 1979-1992.

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