Systematics and phylogeography of the genus Tigriopus (Copepoda, Harpacticoida, Harpacticidae) in the basin of the Mediterranean Sea

Tesis por compendio[ES] El género de copépodos Tigriopus Norman, 1869 se distribuye en todo el mundo en piscinas costeras de rocas y actualmente se considera que incluye 15 especies válidas. Tigriopus fulvus (Fischer, 1860), con su subespecie Tigriopus fulvus adriaticus Van Douwe 1913 y Tigriopus fulvus algiricus Monard 1935, actualmente se informa que ocurren en el área mediterránea, pero actualmente se desconoce la diversidad real del género. Nuestro objetivo fue evaluar la identidad real de las poblaciones mediterráneas de Tigriopus y dilucidar su taxonomía y patrón de diversidad genética. Para alcanzar estos objetivos, utilizamos dos enfoques diferentes. El primero, basado en la morfología, donde las posibles diferencias morfológicas entre muestras topotípicas de Tigriopus fulvus s.s. y muestras topotípicas de las dos subespecies fueron investigadas. Un segundo, sobre una base molecular, donde se secuenciaron fragmentos de dos genes de ADN mitocondrial (subunidad I de citocromo c oxidasa, COI y subunidad de ARN ribosómico pequeño, 12S) y un gen de ADN nuclear (28S) para ser usados como marcador de referencia. En este marco, se utilizaron enfoques taxonómicos moleculares, como el descubrimiento automático de brechas de código de barras (ABGD), bPTP (procesos de árbol de Poisson bayesianos) y la relación K / ¿, para investigar la existencia de las supuestas subespecies de Tigriopus a través de la identificación de Operacional Unidades Taxonómicas (OTU). Nuestros datos sugieren la presencia de una sola especie caracterizada por una notable estructura genética basada en la geografía en toda el área de estudio. El patrón de diversidad observado se atribuye tentativamente aquí a una fuerte monopolización de los estanques de rocas por parte de los primeros inmigrantes que los alcanzan. Sin embargo, tal monopolización se ve interrumpida periódicamente por los eventos de extinción locales, que son frecuentes en los hábitats intrínsecamente inestables de las piscinas de rocas Aquí proponemos para este patrón el nombre de "monopolización periódica" ("Clockwork monopolization").[CA] El género de copépodos Tigriopus Norman, 1869 se distribuix en tot el mon en piscines costeres de roques i actualment se considera que inclou 15 especies valides. Tigriopus fulvus (Fischer, 1860), en la seua subespecie Tigriopus fulvus adriaticus Van Douwe 1913 i Tigriopus fulvus algiricus Monard 1935, actualment s'informa que ocorren en l'area mediterranea, pero actualment se desconeix la diversitat real del género. Nostre objectiu fon evaluar l'identitat real de les poblacions mediterranees de Tigriopus i dilucidar la seua taxonomia i patrón de diversitat genetica. Per a alcançar estos objectius, utilisem dos enfocaments diferents. El primer, basat en la morfologia, a on les possibles diferencies morfologiques entre mostres topotípicas de Tigriopus fulvus s.s. i mostres topotípicas dels dos subespecies foren investigades. Un segon, sobre una base molecular, a on se secuenciaron fragments de dos gens d'adn mitocondrial (subunidad i de citocromo c oxidasa, coi i subunidad d'arn ribosómico menut, 12s) i un gen d'adn nuclear (28s) per a ser amprats com marcador de referencia. En este marc, s'utilisaren enfocaments taxonomics moleculars, com el descobriment automatic de breches de codic de barres (ABGD), bPTP (processos d'arbre de poisson bayesianos) i la relacio K/¿, per a investigar l'existencia de les supostes subespecies de Tigriopus a través de l'identificacio d'operacional unitats taxonomiques (OTU). Nostres senyes sugerixen la presencia d'una sola especie caracterisada per una notable estructura genetica basada en la geografia en tota l'area d'estudi. El patrón de diversitat observat s'atribuix tentativament aci a una forta monopolisacio dels safarejos de roques per part dels primers immigrants que els alcancen. No obstant, tal monopolisacio se veu interrompuda periodicament pels events d'extincio locals, que son freqüents en els hábitats intrinsecament inestables de les piscines de roques aci proponem per a este patrón el nom de "Monopolisacio Periodica" ("Clockwork Monopolization").[EN] The copepod genus Tigriopus Norman, 1869 is distributed worldwide in coastal rock-pools and it is currently considered to include 15 valid species. Tigriopus fulvus (Fischer, 1860), with its subspecies Tigriopus fulvus adriaticus Van Douwe, 1913 and Tigriopus fulvus algiricus Monard, 1935, are currently reported to occur in the Mediterranean area, but the actual diversity of the genus is currently unknown. We aimed to assess the actual identity of Mediterranean Tigriopus populations and to elucidate their taxonomy and pattern of genetic diversity. In order to reach these goals, we use two different approaches. The first, based on morphology, where the possible morphological differences among topotypical samples of Tigriopus fulvus s.s. and topotypical samples of the two subspecies were investigated. A second, on a molecular basis, where fragments of two mitochondrial DNA genes (cytochrome c oxidase subunit I, COI and small ribosomal RNA subunit, 12S) and a nuclear DNA gene (28S) were sequenced to be used as a reference marker. In this frame, molecular taxonomical approaches, such as Automatic Barcode Gap Discovery (ABGD), bPTP (bayesian Poisson Tree Processes) and K/¿ ratio, were used in order to investigate the existences of the alleged subspecies of Tigriopus through the identification of Operational Taxonomic Units (OTUs). Our data suggest the presence of a single species characterized by a noteworthy geographically-based genetic structure in the whole study area. The observed diversity pattern is tentatively ascribed here to a strong monopolization of the rock pools by the first immigrants that reach them. However, such a monopolization is periodically disrupted by local extinction events, which are frequent in the intrinsically unstable rock pool habitats. We propose the name "clockwork monopolization" for this pattern.Vecchioni, L. (2020). Systematics and phylogeography of the genus Tigriopus (Copepoda, Harpacticoida, Harpacticidae) in the basin of the Mediterranean Sea [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/144122TESISCompendi

An account on the taxonomy and molecular diversity of a marine rock-pool dweller, Tigriopus fulvus (Copepoda, Harpacticoida)

The copepod genus Tigriopus Norman, 1869 is distributed worldwide in coastal rock pools and it is currently considered to include 14 valid species. Tigriopus fulvus (Fischer 1860), with its subspecies Tigriopus fulvus adriaticus Van Douwe 1913 and Tigriopus fulvusalgiricus Monard 1935, and Tigriopus minutus Bozic 1960 are currently reported to occur in the Mediterranean area, but the actual diversity of the genus is currently unknown. We aimed to assess the actual identity of Mediterranean Tigriopus populations and to elucidate their taxonomy and pattern of genetic diversity. In order to reach these goals, a fragment of a mitochondrial DNA gene (cytochrome c oxidase subunit I, COI) was sequenced to be used as a reference marker. Our data suggest the presence of a single species characterized by a noteworthy geographi-cally based genetic structure in the whole study area. The observed diversity pattern is tentatively ascribed here to a strong monopolization of the rock pools by the first immigrants that reached them. However, such a monopolization is periodically disrupted by local extinction events, which are frequent in the intrinsically unstable rock pool habitats. We propose the name “clockwork monopolization” for this pattern.El género de copépodos Tigriopus Norman, 1869 se distribuye en todo el mundo en charcas de rocas costeras y se considera que actualmente incluye 14 especies válidas. Tigriopus fulvus (Fischer 1860), con sus subespecies Tigriopus fulvus adriaticus Van Douwe 1913 y Tigriopus fulvus algiricus Monard 1935, y Tigriopus minutus Bozic 1960 han sido descritos para el área del Mediterráneo, pero la diversidad real del género es desconocida actualmente. El objetivo de este estudio fue evaluar la identidad real de las poblaciones mediterráneas de Tigriopus y dilucidar su taxonomía y patrón de diversidad genética. Con este fin, se secuenció un fragmento del gen de ADN mitocondrial (citocromo c oxidasa subunidad I, COI) como marcador de referencia. Los resultados sugieren la presencia de una sola especie caracterizada por una estructuración genética con una notable base geográfica en toda el área de estudio. El patrón de diversidad observado aquí se atribuye tentativamente a una fuerte monopolización de las charcas de las costas rocosas por parte de los primeros inmigrantes que las alcanzan. Sin embargo, tal monopolización se interrumpe periódicamente por los eventos de extinción local, los cuales son frecuentes en los hábitats de charcas de rocas que son intrínsecamente inestables. Aquí proponemos para este patrón el nombre de “monopolización periódica” (“clockwork monopolization”)

On the occurrence and distribution of Calanipeda aquaedulcis Kritschagin, 1873 (Copepoda, Calanoida, Pseudodiaptomidae) in Sicily, Italy, with some notes on coexistence and species replacement in calanoid copepods

The only population of the pseudodiaptomid copepod Calanipeda aquaedulcis Kritschagin, 1873 to date reported to occur in Sicily disappeared at the beginning of the XXI century due to deep environmental changes which affected the single site (Lake Biviere di Gela) known for this species on the island. In that site C. aquaedulcis is now replaced by Copidodiaptomus numidicus (Gurney, 1909), a diaptomid copepod whose distribution has been greatly increasing since the second half of the last century. In the present note, the occurrence of C. aquaedulcis in 12 novel water bodies spread throughout Sicily is reported, and some environmental data on the sites where the species was collected are provided. Moreover, in a few sites C. aquaedulcis was found to co-occur with Copidodiaptomus numidicus. The recorded co-occurrence of these two species and the replacement of C. aquaedulcis with C. numidicus in Lake Biviere di Gela are briefly discussed

Range‐wide phylogeography and taxonomy of the marine rock pools dweller Tigriopus fulvus (Fischer, 1860) (Copepoda, Harpacticoida)

In the light of the wide distribution and ecological importance of the genus Tigriopus in coastal rock pool habitats, and of its frequent use in aquaculture and as a model organism, we investigated the identity of the Atlantic–Mediterranean Tigriopus populations and elucidated their taxonomy and patterns of morphological and genetic diversity. In order to reach these goals, an "integrative taxonomy" multisource approach was implemented. First, we investigated the constancy and taxonomical value of the morphological characters currently used to distinguish among the Tigriopus species occurring in the Mediterranean and in Eastern Atlantic area north of the Tropic of Cancer, and checked the actual morphological differences possibly present among topotypical samples of Tigriopus fulvus fulvus (Fischer, 1860) and its two subspecies Tigriopus fulvus adriaticus Van Douwe, 1913 and Tigriopus fulvus algiricus Monard, 1935. Then, we sequenced fragments of mitochondrial (12S) and nuclear (28S) genes. In the frame of this study, different "DNA taxonomy" approaches were implemented in order to check whether the subspecies of Tigriopus fulvus were actually lineages evolving independently, that is, valid species according to the "evolutionary genetic species concept." The results coherently indicate the presence of a single species, characterized by constant morphology and a noteworthy geographically based genetic structure in the whole study area. No morphological or genetic support was found for the taxa of allegedly subspecific rank within T. fulvus, which are thus to be considered junior synonyms of T. fulvus s.s. Finally, a restricted locus typicus is established for T. fulvus, and a neotype is designated

Una revisión de la taxonomía y diversidad molecular de un habitante de las charcas litorales Tigriopus fulvus (Copepoda, Harpacticoida)

[ES] El género de copépodos Tigriopus Norman, 1869 se distribuye en todo el mundo en charcas de rocas costeras y se considera que actualmente incluye 14 especies válidas. Tigriopus fulvus (Fischer 1860), con sus subespecies Tigriopus fulvus adriaticus Van Douwe 1913 y Tigriopus fulvus algiricus Monard 1935, y Tigriopus minutus Bozic 1960 han sido descritos para el área del Mediterráneo, pero la diversidad real del género es desconocida actualmente. El objetivo de este estudio fue evaluar la identidad real de las poblaciones mediterráneas de Tigriopus y dilucidar su taxonomía y patrón de diversidad genética. Con este fin, se secuenció un fragmento del gen de ADN mitocondrial (citocromo c oxidasa subunidad I, COI) como marcador de referencia. Los resultados sugieren la presencia de una sola especie caracterizada por una estructuración genética con una notable base geográfica en toda el área de estudio. El patrón de diversidad observado aquí se atribuye tentativamente a una fuerte monopolización de las charcas de las costas rocosas por parte de los primeros inmigrantes que las alcanzan. Sin embargo, tal monopolización se interrumpe periódicamente por los eventos de extinción local, los cuales son frecuentes en los hábitats de charcas de rocas que son intrínsecamente inestables. Aquí proponemos para este patrón el nombre de ¿monopolización periódica¿ (¿clockwork monopolization¿).[EN] The copepod genus Tigriopus Norman, 1869 is distributed worldwide in coastal rock pools and it is currently considered to include 14 valid species. Tigriopus fulvus (Fischer 1860), with its subspecies Tigriopus fulvus adriaticus Van Douwe 1913 and Tigriopus fulvus algiricus Monard 1935, and Tigriopus minutus Bozic 1960 are currently reported to occur in the Mediterranean area, but the actual diversity of the genus is currently unknown. We aimed to assess the actual identity of Mediterranean Tigriopus populations and to elucidate their taxonomy and pattern of genetic diversity. In order to reach these goals, a fragment of a mitochondrial DNA gene (cytochrome c oxidase subunit I, COI) was sequenced to be used as a reference marker. Our data suggest the presence of a single species characterized by a noteworthy geographically based genetic structure in the whole study area. The observed diversity pattern is tentatively ascribed here to a strong monopolization of the rock pools by the first immigrants that reached them. However, such a monopolization is periodically disrupted by local extinction events, which are frequent in the intrinsically unstable rock pool habitats. We propose the name "clockwork monopolization" for this pattern.We wish to thank Prof. Vezio Cottarelli for the help he provided us with the collection of some samples of T. fulvus and for the stimulating discussions we had on the topic of this manuscript. G Alfonso (University of Salento, Italy) and A Millán (University of Murcia, Spain) kindly provided some of the studied samples.Vecchioni, L.; Marrone, F.; Rodilla, M.; Belda, E.; Arculeo, M. (2019). An account on the taxonomy and molecular diversity of a marine rock-pool dweller, Tigriopus fulvus (Copepoda, Harpacticoida). Ciencias Marinas. 45(2):59-75. https://doi.org/10.7773/cm.v45i2.2946S597545

Unveiling the egg microbiota of the loggerhead sea turtle <i>Caretta caretta</i> in nesting beaches of the Mediterranean Sea

Microbes have central roles in the development and health of animals, being the introduction of specific microbial species a potential conservation strategy to protect animals from emerging diseases. Thus, insight into the microbiota of the species and their habitats is essential. In this manuscript, we report for the first time the bacterial composition of all the components (eggshells of hatched and unhatched eggs, internal content of unhatched eggs, intestinal content of hatchling and pipping sea turtles, and sand) of three nesting beaches of Caretta caretta along the Italian coasts of the Mediterranean Sea. The analysis of 26 amplicon samples was carried out using next-generation sequencing analysis, targeting V3-V4 regions of the bacterial 16S rRNA gene. Samples featured mainly Proteobacteria, Actinobacteria, Firmicutes, and Bacteroidetes, whose percentages depended on the sample type. Our results showed that, although from different sampling sites, the internal content of the unhatched eggs, intestinal content of hatchling and pipping sea turtles share the microbiota, which was yet different from that of eggshells and sand of the same nesting beach. This study suggests the maternal and environmental influence alongside a protective role of eggshells in shaping the egg microbiota of Caretta caretta sea turtles

Distribution and genetic lineages of the Craspedacusta sowerbii species complex (Cnidaria, Olindiidae) in Italy

Olindiid freshwater jellyfishes of the genus Craspedacusta Lankester, 1880 are native to eastern Asia; however, some species within the genus have been introduced worldwide and are nowadays present in all continents except Antarctica. To date, there is no consensus regarding the taxonomy within the genus Craspedacusta due to the morphological plasticity of the medusa stages. The species Craspedacusta sowerbii Lankester, 1880 was first recorded in Italy in 1946, and until 2017, sightings of the jellyfish Craspedacusta were reported for 40 water bodies. Here, we shed new light on the presence of the freshwater jellyfishes belonging to the genus Craspedacusta across the Italian peninsula, Sardinia, and Sicily. First, we report 21 new observations of this non-native taxon, of which eighteen refer to medusae sightings, two to environmental DNA sequencing, and one to the finding of polyps. Then, we investigate the molecular diversity of collected Craspedacusta specimens, using a Bayesian analysis of sequences of the mitochondrial gene encoding for Cytochrome c Oxidase Subunit I (mtDNA COI). Our molecular analysis shows the presence of two distinctive genetic lineages: (i) a group that comprises sequences obtained from populations ranging from central to northern Italy; (ii) a group that comprises three populations from northern Italy—i.e., those from the Lake Levico, the Lake Santo of Monte Terlago, and the Lake Endine—and the single known Sicilian population. We also report for the first time a mtDNA COI sequence obtained from a Craspedacusta medusa collected in Spai

New distributional data for the Mediterranean medicinal leech Hirudo verbana Carena, 1820 (Hirudinea, Hirudinidae) in Italy, with a note on its feeding on amphibians

Scarce data are currently available about the distribution of the Mediterranean medicinal leech Hirudo verbana in Italy, and most of the known occurrence localities are based on records collected in the nineteenth and the first half of the twentieth century, which were not confirmed in the last decades, mostly due to a lack of surveys. Accordingly, the available knowledge on the distribution of the species is far from being updated and representative, although a significant negative trend of H. verbana throughout the country is supposed. The lack of sound distribution data is a significant shortfall, which hinders the implementation of the monitoring of the species as required by the Article 17 of the “Habitats Directive” on the species of Union concern. To provide recent, validated distributional data for the Mediterranean medicinal leech in Italy to be used as baseline data for further surveys and monitoring, we present herein a set of unpublished observations collected in the last decades in peninsular Italy, Sicily, and Sardinia. Moreover, we report observation of H. verbana feeding on amphibians, a feeding habit to date poorly documented for the Mediterranean medicinal leech. Based on both published and novel data, H. verbana is characterised by a large but fragmented distribution in Italy. Therefore, dedicated monitoring programs and the collection of validated occasional observations are highly desirable to get a clearer picture of the real distribution of the species

Ridurre lo scarto nella pesca con reti da posta: il caso di studio nella AMP delle isole Egadi

Sebbene la selettività nel tramaglio sia assodata, rimangono solitamente catturati dalla rete individui di specie bentoniche che non hanno rilevanza ai fini della commercializzazione ma svolgono ruoli ecologici rilevanti nell'ecosistema marino. Oltre che il danno ecologico, la cattura di invertebrati del benthos costituisce un aggravio di lavoro per i pescatori che impiegano molto tempo per la pulizia delle reti. L'obiettivo generale di questo lavoro è la riduzione della cattura "indesiderata" di invertebrati del benthos, migliorando la selettività ed aumentando ulteriormente la sostenibilità della pesca con il tramaglio. Nello specifico si è voluto sperimentare l'installazione di una "greca" sulla parte terminale del tramaglio. Si tratta di una fascia di rete mono-panno montata alla base dell'attrezzo, prima della lima dei piombi, allo scopo di minimizzare la cattura di specie accessorie e il rischio di danneggiamento degli organismi bentonici. Sono state impiegate 2 imbarcazioni provviste di attrezzo sperimentale in ciascuna delle tre differenti aree di indagine (Favignana, Marettimo, Trapani), per un totale di 48 uscite. Le prove di pesca sono state condotte all’interno dell’Area Marina Protetta Isole Egadi. E’ stato previsto che ogni barca impieghi un attrezzo sperimentale di 1000 metri di lunghezza in cui si alternano 50 metri di tramaglio standard (pannello interno con maglia di 31,25 mm di lato, pannelli esterni con maglia di 180 mm di lato), con pezze delle stesse caratteristiche a cui è stata aggiunta una “greca” di 35 cm di altezza, di maglia di 50 mm di lato. Dal confronto delle catture di specie commerciali e non commerciali ottenute con il tramaglio sperimentale, valutando le differenze di cattura tra le pezze con "greca" e quelle armate in maniera tradizionale, si è osservato che si ha una riduzione degli organismi bentonici nonché delle specie commerciali. L'attrezzo risulta in generale più selettivo nei confronti della rete tradizionale