Morphological and genetic analyses of the first record of the Niger Hind, Cephalopholis nigri (Perciformes: Serranidae), in the Mediterranean Sea and of the African Hind, Cephalopholis taeniops, in Malta
© The Author(s) 2016
Received: 8 September 2016
Accepted: 17 November 2016
Published: 24 November 2016
Non-native marine species, including tropical eastern Atlantic fish species are on the increase in Malta, with shipping activities being the main vector for the movement of these alien species from the Atlantic into the Mediterranean Sea. This calls for cooperation and collaboration between various sea-users and researchers to ensure continuous monitoring of coastal biodiversity.
Research methods involving local fishermen cooperation in monitoring efforts to identify and track populations of alien species in the Central Mediterranean has led to new records for the genus Cephalopholis (Perciformes: Serranidae) in Malta. Morphological characteristics, meristic counts and mitochondrial DNA sequences from specimens of both species sampled from Maltese waters were analysed to confirm their species identify accurately, essential for tracking their respective population expansions in the Mediterranean.
Results and conclusion
Results from this study have led to confirmation of the first record of the Niger Hind, Cephalopholis nigri (Günther, 1859), in the Mediterranean Sea and of the establishment of the African Hind, Cephalopholis taeniops (Valenciennes, 1828) in Maltese waters.
KeywordsCephalopholis nigri Cephalopholis taeniops Non-native New record Mediterranean
Groupers are economically important species and are caught by commercial, artisanal and recreational fishermen. In the Mediterranean, the subfamily Epinephelinae is represented by six native species, Epinephelus aeneus, E. caninus, E. costae, E. marginatus, Hyporthodus haifensis and Mycteroperca rubra (Heemstra and Randall 1993; Froese and Pauly 2016). In addition to these, there are non-native tropical Epinephelinae species, including six Indo-Pacific Epinephelus species, E. malabaricus, E. coioides, E. merra, E. fasciatus, E. geoffroyi and E. areolatus, that were first recorded in 1966 (Heemstra and Randall 1993), 1969 (Ben-Tuvia and Lourie 1969; Heemstra and Golani 1993), 2004 (Lelong 2005), 2011 (Bariche and Heemstra 2012), 2015 (Golani et al., 2015) and 2015 (Rothman et al., 2016) respectively. Additionally, another two Atlantic species, Cephalopholis taeniops, first noted in 2002 (Ben Abdallah et al., 2007), and Mycteroperca fusca, reported in 2010 (Heemstra et al., 2010) were also found in the Mediterranean Sea. The increase in new records of non-native Epinephelinae species in this region follows the trend noted for a number of other tropical fish groups (Golani 2010; Golani 2013; Vella et al., 2015a, b & 2016a).
The genus Cephalopholis Bloch & Schneider, 1801 is composed of 24 species, 19 of which occur in the Red Sea and the Indo-Pacific region, one is from the eastern Pacific, two from the western Atlantic and two from the eastern Atlantic (Heemstra and Randall 1993; Froese and Pauly 2016). These species are primarily tropical and subtropical species, none of which are native to the Mediterranean Sea. The two species analysed in this study are C. nigri and C. taeniops, both of which are of tropical eastern Atlantic Ocean origin, with their native range extending from Angola to the Canary Islands and Western Sahara respectively (Heemstra and Randall 1993; Froese and Pauly 2016). While specimens of C. taeniops have been morphologically analysed in Libya (Ben Abdallah et al., 2007) and Israel (Salameh et al., 2009), prior to this study there have been no records of C. nigri in the Mediterranean Sea.
Results and discussion
Measurements and meristic counts of the first record of Cephalopholis nigri in the Mediterranean Sea and of a Cephalopholis taeniops both specimens caught from Maltese waters
Maximum body depth
Length of dorsal fin base
Pectoral fin base
Anal fin base
Dorsal fin spines
Dorsal fin soft rays
Pectoral fin soft rays
Pelvic fin spines
Pelvic fin soft rays
Anal fin spines
Anal fin soft rays
Scales on lateral line
A total of 2026 bp were sequenced from the mtDNA of both specimens. The sequence lengths obtained were 412 bp, 604 bp, 585 bp and 425 bp for cytochrome b (Cytb), cytochrome c oxidase I (COI), 16S rRNA (16S) and 12S rRNA (12S) genes respectively. Each sequence was run via BLASTn to identify sequence matches.
The two studied specimens were genetically confirmed to species level at the 12S and the 16S genes at >99.3% matches with Craig and Hastings (2007). The 12S matched AY949451 and AY949387, while 16S matched AY947604 and AY947589 for C. nigri and C. taeniops respectively. Cytochrome B gene of C. taeniops also confirmed the species with a 100% match to EF455990-1 specimens from Mauritania (Gonzalez-Sevilla et al., unpublished). It was not possible to compare CytB in C. nigri and COI for both species with any other sequences as there is no publically available data for them. This study presents the first sequences for these alien species collected from the Mediterranean Sea.
In December, 2015, an additional three specimens of C. taeniops have been caught in close proximity to each other (Fig. 4). The three recorded individuals included one specimen with a colouration similar to the one described in Fig. 2, and another two specimens of the less common darker variety (Seret, 1981), one of which can be seen in Fig. 4.
The number of alien species in the Mediterranean Sea are on the increase (Golani 2010; 2013), including species of both Atlantic and Indo-Pacific origin. In fact the list of non-native species extends across several taxa, including groupers from the family Epinephelinae. Cephalopholis nigri is the latest record of a non-native grouper in the region. The species is not known to be a natural migrant as there are no records of this species elsewhere in the Mediterranean Sea. Moreover, given that Epinephelinae species are highly prized catches, it is very unlikely that records of this species would have gone unnoticed by divers and fishermen. This first Mediterranean record of C. nigri is from a busy Maltese harbour which caters for dockyards, oil platform servicing, transhipment activities and berthing of large marine vessels including cruise-liners and super-yachts. Like elsewhere in the region (Galil, 2006; Katsanevakis et al., 2014), such maritime activity can be considered as the main vector in the introduction of alien species. This new record of alien species, follows others from areas that are characterized by intensive marine activity in Malta, such as the first Mediterranean records of Stegastes variabilis, Lutjanus fulviflamma and Abudefduf hoefleri (Vella et al., 2015a, b & 2016a). Nonetheless, one cannot exclude the possibility that C. nigri was an aquarium release given that members of the genus Cephalopholis are exported as ornamental fish (Monteiro-Neto et al., 2003) and in recent years this industry has led to an increase in alien species within the Mediterranean Sea (Guidetti et al., 2016; Zenetos et al., 2016).
Prior to this study there have only been isolated sighting reports for C. taeniops in Maltese waters without the analyses of any voucher specimen for this alien grouper species. This study has therefore presented the first scientific morphometric and genetic analyses of this species from Maltese waters, while reporting an increasing number of sightings and photographic records. The four new records of C. taeniops indicate the establishment of a population in Maltese waters. Ongoing scientific monitoring would be required to further study the expanding range of this species, although the permanence of its population is threatened by the local fishing industry that is looking at C. taeniops as another grouper that may be exploited for commercial means (pers. comm. with fishermen). This latter activity can prove to be beneficial in controlling the spread of this non-native species.
The occurrence and proliferation of these carnivorous species need monitoring as one cannot exclude the possibility that expanding populations of these species might lead to interspecific competition for resources with other already vulnerable native Epinephelinae species in the Mediterranean and in Maltese waters. However, since groupers are species under pressure from over-exploitation these alien species are also being exploited for local consumption, keeping their numbers low.
The diagnostic features used in the morphological identification of both specimens followed Heemstra and Randall (1993) and Froese and Pauly (2016). All length measurements were taken to the nearest 0.1 mm using electronic calipers and mass was recorded to the nearest 0.01 g.
DNA was extracted using GF-1 Tissue DNA Extraction Kit (Vivantis Technologies). PCR amplifications were carried out for Cytb, COI and 16S using the primers sets as described in Vella et al., (2016a, b), and for the 12S using H1478 and L1091 primers (Kocher et al., 1989). PCR amplifications were carried out following the amplification protocols described in Vella et al., (2016a). PCR products were purified and sequenced via ABI3730XL sequencer using both the forward and reverse primers. The sequences, at both nucleotide and amino acid level, were analyzed using Geneious v6 (http://www.geneious.com, Kearse et al., 2012). The sequences obtained were deposited in GenBank, with accession numbers of C. taeniops KX758563-6 and C. nigri KX758567-70 for Cytb, COI, 16S and 12S respectively. These sequences were compared to other sequences available in genomic databases using BLASTn.
12S rRNA gene
16S rRNA gene
Cytochrome c oxidase I gene
Cytochrome b gene
Thanks are due to sport fishermen, in particular to Mr. R. Farrugia, Hooked on Fishing sports club and Mr. J. Tanti who have assisted in this research.
The genetic research disclosed in this publication is funded by the REACH HIGH Scholars Programme-Post Doctoral Grants. The grant may be part-financed by the European Union, Operational Programme II - Cohesion Policy 2014–2020 “Investing in human capital to create more opportunities and promote the well being of society” - European Social Fund.
Availability of data and materials
The genetic sequence data supporting the results of this article are available in GenBank, as accession numbers KX758563-6 (C. taeniops) and KX758567-70 (C. nigri) for Cytb, COI, 16S and 12S genes respectively. While the voucher specimens CBRG/F.160710/CN001 and CBRG/F.160409/CT001 were collected from Malta and have been deposited in the ichthyological collection of the Conservation Biology Research Group laboratory at the University of Malta.
NV and AV have contributed to all aspects of the research work presented here including the conception and design of the molecular genetics research, analyses and interpretation of both genetic and morphological data and were involved in finalizing the manuscript. SAD contributed to the morphological research work of the specimens collected. All three authors were involved in the drafting of the manuscript and gave approval for publication.
All three authors are researchers of the Conservation Biology Research Group, Department of Biology, University of Malta.
None of the authors of this paper have financial or non-financial competing interests associated with this research work.
Consent for publication
Authors gave consent to publish this MS.
Ethics approval and consent to participate
This research did not require any ethics approval or consent to participate.
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