The authentication of food products and the verification of their identity are of major importance for consumers. Food fraud through mislabeling is an illegal practice consisting of the substitution of an expensive food product by a relatively cheaper one, misleading false labelling of their origin and adulteration in processed or frozen products. This issue is particularly of high importance concerning fish and seafood, which are easily adulterated primarily due to difficult morphological identification. Fish species of the Mullidae family are considered among the most high-valued seafood products traded in Greece and Eastern Mediterranean in general, in terms of the price and demand. Specifically, the red mullet (Mullus barbatus) and the striped red mullet (Mullus surmuletus) are both indigenous in the Aegean (FAO Division 37.3.1) and the Ionian (FAO Division 37.2.2) Seas, with high levels of consumers’ preferences. However, they could be easily adulterated or misidentified by the invasive Aegean Sea Lessepsian migrator goldband goatfish (Upeneus moluccensis) as well as by the imported West African goatfish (Pseudupeneus prayensis). Keeping this in mind, we designed two novel, time-saving and easy-to-apply multiplex PCR assays and one multiple Melt–Curve analysis real-time PCR for the identification of these four species. These methodologies are based on species-specific primers targeting single nucleotide polymorphisms (SNPs) detected via sequencing analysis of the mitochondrial cytochrome C oxidase subunit I (CO1) and of the cytochrome b (CYTB) genes in newly collected individuals, with additional comparison with congeneric and conspecific haplotypes obtained from the GenBank database. Both methodologies, targeting CO1 or CYTB, utilize one common and four diagnostic primers, producing amplicons of different length that are easily and reliably separated on agarose gel electrophoresis, yielding a single clear band of diagnostic size for each species or a certain Melt–Curve profile. The applicability of this cost-effective and fast methodology was tested in 328 collected specimens, including 10 cooked samples obtained from restaurants. In the vast majority (327 out of the 328) of the specimens tested, one single band was produced, in agreement with the expected products with a single exception a M. barbatus sample that was identified as M. surmuletus, the identity of which was confirmed using sequencing, indicating erroneous morphological identification. The developed methodologies are expected to contribute to the detection of commercial fraud in fish authentication.

Admixture and hybridization may play a key role in population dynamics and speciation with respect to habitat, demographic history, and adaptive selection. The present study examines the genetic diversity of two congeneric—but in different subgenera—holothurians that live in sympatry in mixed populations. Strong evidence of admixture was provided by analyzing RAD sequencing data from 90 and 67 individuals of Holothuria (Holothuria) tubulosa and Holothuria (Roweothuria) poli, respectively, from various areas of the Hellenic Seas (eastern Mediterranean). Coalescent demographic analysis revealed a recent directional gene flow from H. poli to H. tubulosa. The two species populations diverged around 13.5 thousand years ago, just after the Last Glacial Maximum. According to the pairwise sequential Markovian coalescent approach, the historical population effective sizes for both species declined during the last Pleistocene glaciations, probably due to population decline, followed by a relative rapid recovery as it is calculated using LD methods. The presented results imply a role for admixture upon secondary contact and are consistent with the recent suggestion that the genomic underpinning of ecological speciation often has an older, allopatric origin.

The red porgy (Pagrus pagrus) and the common dentex (Dentex dentex) are Sparidae species of high commercial value, traded in the Greek market. In some cases, fish species identification from Greek fisheries is difficult for the consumer due to the strong morphological similarities with their imported counterparts or closely related species such as Pagrus major, Pagrus caeroleustictus, Dentex gibbosus and Pagellus erythrinus, especially when specimens are frozen, filleted or cooked. Techniques based on DNA sequencing, such as COI barcoding, accurately identify species substitution incidents; however, they are time consuming and expensive. In this study, regions of mtDNA were analyzed with RFLPs, multiplex PCR and HRM in order to develop a rapid method for species identification within the Sparidae family. HRM analysis of a 113 bp region of cytb and/or a 156 bp region of 16s could discriminate raw or cooked samples of P. pagrus and D. dentex from the aforementioned closely related species and P. pagrus specimens sampled in the Mediterranean Sea when compared to those fished in the eastern Atlantic. HRM analysis exhibited high accuracy and repeatability, revealing incidents of mislabeling. Multiple samples can be analyzed within three hours, rendering this method a useful tool in fish fraud monitoring.

The Mediterranean monk seal (Monachus monachus) is classified by the IUCN as “endangered,” with a global population estimated to number fewer than 800 individuals. Our understanding of the biology and health status of the species is still limited, rendering every medical case a challenge for conservationists and veterinary clinicians. Although studying and managing disease in wild marine hosts is complex and challenging, studying and mitigating the effects of any disease to the Mediterranean monk seal is of utmost importance for conservation. The aim of this study was to document for the first time the presence of the hookworm Uncinaria hamiltoni in rehabilitated Mediterranean monk seal pups in Greece. A detailed examination protocol was followed for all pups that live-stranded over 30 years in 22 different locations, including physical, parasitological, and other examinations. Hookworms (adults and/or eggs) were detected in all the fecal samples, from all animals. Molecular identification using MtDNA (COI) and ribosomal DNA (D2/D3 28S and internal transcribed spacer [ITS] regions) identified the nematode species as Uncinaria hamiltoni. The clinical impacts and the benefits of anthelmintic treatment as a tool for the conservation management of the species are discussed.

Coalescent methods in population genetics aim to detect biodiversity patterns, evolutionary mechanisms, and signatures of historical changes in effective population sizes with respect to the species fidelity. Restriction site-associated DNA sequencing (RADseq) was used to evaluate the population dynamics of invertebrate species within the same localities. New sequencing technologies, such as the ones employed by population genetics, could be used to improve the management and sustainability of marine and aquaculture resources. Sea cucumbers (Holothuria tubolosa) showed genetic differentiation patterns favoring limited gene flow between studied areas. Similar results for clams (Venus verrucosa) suggest local adaptation and low-dispersal abilities for sessile organisms. On the contrary, cuttlefish (Sepia officinalis) exhibited a panmictic pattern, resulting in a single genetic stock in the area. The larvae settlement duration may be responsible for such interspecies variations. Interspecies demographic modeling revealed different environmental pressures of historical events’ signatures with respect to the three invertebrates. Sea cucumbers favor a post-glacial bottleneck event followed by a more recent recovery, whereas cuttlefish favor an expansion before the late glacial maximum. Lastly, clams showed a constant effective population size in the area. The results of historical demographic changes in natural populations provide opportunities for critical evaluation and management in terms of the conservation of the species in the area.

The Mediterranean monk seal (Monachus monachus) is classified as an endangered species by the IUCN, with a global population that does not exceed 800 individuals. There is limited understanding around the biology and health status of the species, rendering possible parasitic infections grave for its conservation efforts. The aim of the current study was the molecular identification of a parasitic nematode found in the digestive system of a sub-adult Mediterranean monk seal individual, that was found stranded in the area of Pagasitikos Gulf, Greece in 2019. Analysis of the stomach contents revealed the presence of two intact female nematode individuals. Standard protocols were followed as DNA extraction of the parasites was conducted and PCR amplification of the cytochrome oxidase subunit I (COI) mitochondrial gene was implemented. Sequencing analysis of a 585 bp-amplified product displayed a 96% similarity of the screened nematodes to the Pseudoterranova bulbosa species. Bayesian inference was implemented for the subsequent tree reconstruction. The phylogenetic tree revealed a clear genetic similarity between our parasitic nematode individuals named as Pseudoterranova spp. and Pseudoterranova bulbosa (bootstrap value: 82%), which is indicated for the first and only time as such, to be found in the waters of the Mediterranean Sea and also in the stomach of a Mediterranean monk seal.