Paolo Merella

Diet of the european flying squid Todarodes sagittatus (Cephalopoda : Ommastrephidae) in the Balearic sea (western Mediterranean)

Stomach contents of Todarodes sagittatus caught by trawlers working from 100 to 800 m depth in the Balearic Sea (western Mediterranean) were studied. From the 348 stomachs examined (153 males and 195 females) 33.62% were empty (39.21% in males and 29.74% in females). The diet of the squid was composed of 58 different prey items belonging to four major groups: Osteichthya, Crustacea, Cephalopoda and Chondrichthya. Osteichthyes, crustaceans and cephalopods were the most common prey, with a frequency of occurrence value of 84.85, 48.92 and 29.87% respectively. A change in the diet as the squid grows was observed, since juveniles feed basically on fishes while adults prey more actively on crustaceans. Analysis of the diet by size-classes reflected an ontogenetic migration to deeper waters since, parallel to the increase of size, a raise in the percentage of prey species inhabiting deeper waters was detected. Cannibalism was quite frequent, since T. sagittatus was the second most common cephalopod prey. Females had higher fullness-weight index and lower emptiness index than males, which reflects their major energetic demand for egg production.

Prevalence of Anisakis spp. and Hysterothylacium spp. larvae in teleosts and cephalopods sampled from waters off Sardinia.

A study was carried out on the presence of Anisakis and Hysterothylacium larvae in fish and cephalopods caught in Sardinian waters. A total of 369 specimens of 24 different species of teleosts and 5 species of cephalopods were collected from different fishing areas of Sardinia. Larvae were detected and isolated by both visual inspection and enzymatic digestion. These methods allowed Anisakis type I and type II third-stage larvae and Hysterothylacium third- and fourth-stage larvae to be detected. The prevalence, mean intensity, and mean abundance were calculated. The results obtained showed the highest prevalence of Anisakidae in Zeus faber (100%) and of Anisakis in Micromesistius poutassou (87.5%). The highest prevalence of Anisakis type I larvae was in M. poutassou (81.2%), and that of Anisakis type II larvae was in Todarodes sagittatus (20%). The highest values for prevalence, mean intensity, and mean abundance for Hysterothylacium were found in Z. faber. These prevalences and the mean intensity and abundance were higher than those reported by different authors in other Mediterranean areas. This may be because the enzymatic digestive method used in this research resulted in higher recovery levels. The data suggest that Sardinia may be a high-risk area for zoonotic diseases and that measures such as information campaigns, aimed at both sanitary service personnel and consumers, should be employed to limit the spread of such zoonosis.

Molecular characterization of Fasciola spp. from the endemic area of northern Iran based on nuclear ribosomal DNA sequences.

Fasciolosis caused by Fasciola spp. (Platyhelminthes: Trematoda: Digenea) is considered as the most important helminth infection of ruminants in tropical countries, causing considerable socioeconomic problems. In the endemic regions of the North of Iran, Fasciola hepatica and Fasciola gigantica have been previously characterized on the basis of morphometric differences, but the use of molecular markers is necessary to distinguish exactly between species and intermediate forms. Samples from buffaloes and goats from different localities of northern Iran were identified morphologically and then genetically characterized by sequences of the first (ITS-1) and second (ITS-2) Internal Transcribed Spacers (ITS) of nuclear ribosomal DNA (rDNA). Comparison of the ITS of the northern Iranian samples with sequences of Fasciola spp. from GenBank showed that the examined specimens had sequences identical to those of the most frequent haplotypes of F. hepatica (n=25, 48.1%) and F. gigantica (n=20, 38.45%), which differed from each other in different variable nucleotide positions of ITS region sequences, and their intermediate forms (n=7, 13.45%), which had nucleotides overlapped between the two Fasciola species in all the positions. The ITS sequences from populations of Fasciola isolates in buffaloes and goats had experienced introgression/hybridization as previously reported in isolates from other ruminants and humans. Based on ITS-1 and ITS-2 sequences, flukes are scattered in pure F. hepatica, F. gigantica and intermediate Fasciola clades, revealing that multiple genotypes of Fasciola are able to infect goats and buffaloes in North of Iran. Furthermore, the phylogenetic trees based upon the ITS-1 and ITS-2 sequences showed a close relationship of the Iranian samples with isolates of F. hepatica and F. gigantica from different localities of Africa and Asia. In the present study, the intergenic transcribed spacers ITS-1 and ITS-2 showed to be reliable approaches for the genetic differentiation of Fasciola spp., providing bases for further studies on F. hepatica, F. gigantica and their intermediate forms in the endemic areas in Asia.

Molecular and epidemiological data on Anisakis spp. (Nematoda: Anisakidae) in commercial fish caught off northern Sardinia (western Mediterranean Sea)

Anisakiasis is a fish-borne zoonosis caused by third stage larvae of the nematode Anisakis sp. present in fish or cephalopods. This is the first contribution to the molecular identification and epidemiology of Anisakis spp. in commercial fish from the Gulf of Asinara (Sardinia, western Mediterranean Sea). Between April 2006 to November 2011, 777 specimens of 10 fish species (Engraulis encrasicolus, Merluccius merluccius, Micromesistius poutassou, Phycis blennoides, Sardina pilchardus, Sardinella aurita, Scomber colias, Sphyraena viridensis, Trachurus mediterraneus, Trachurus trachurus) were examined for Anisakis sp. larvae. A total of 1286 larvae were found in 218 fish. The great majority of larvae were located in the body cavity, and only a small part (60, 4.7%) in the muscle. All the Type I larvae (1272) were identified as Anisakis pegreffii and all the Type II (14) as Anisakis physeteris, confirming that A. pegreffii is the dominant species and the most important agent of human anisakiasis in the western Mediterranean Sea.

Genetic characterization of Fasciola hepatica from Tunisia and Algeria based on mitochondrial and nuclear DNA sequences

Fasciolosis caused by Fasciola spp. (Platyhelminthes: Trematoda: Digenea) is considered the most important helminth infection of ruminants in tropical countries, causing considerable socioeconomic problems. In the present study, samples identified morphologically as Fasciola hepatica from sheep and cattle from different geographical locations of Tunisia and Algeria were genetically characterised by sequences of the first (ITS-1), the 5.8S and second (ITS-2) Internal Transcribed Spacers (ITS) of nuclear ribosomal DNA (rDNA) and mitochondrial Cytochrome c Oxidase subunit I (COI) gene. Comparison of the ITS and COI sequences of the North African samples with sequences of Fasciola spp. from GenBank confirmed that all samples from Tunisia and Algeria samples belong to a single species, namely F. hepatica. Several specimens from Tunisia and Algeria showed a substitution C/T in position 859 in the ITS-2 sequences, previously reported from Spain, suggesting that the above mentioned variant may have a common origin and spread recently throughout the three countries because of movement of infected animals. This is the first molecular characterization of F. hepatica in North Africa which provides a foundation for further studies on Fasciola spp. in Tunisia and Algeria.

Molecular Characterization of Anisakis Larvae from Fish Caught Off Sardinia

abstract:  Anisakis spp. larvae are parasitic, and potentially zoonotic, nematodes transmitted by marine fish and cephalopods, which are the main intermediate hosts of the third larval stage. The accidental consumption of infected raw or poorly cooked fish may cause gastroenteric diseases and allergies in humans. The aim of the present study was to use polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) to define the occurrence, species variability, and host preferences of Anisakis spp. larvae in fish caught off the coast of Sardinia. Necropsy was used on 285 samples; 552 Anisakis spp. L3 larvae were isolated from 87 fish that tested positive for this nematode. Anisakis pegreffii was most frequently encountered (90.6%), with a primary preference for Scomber scombrus, Zeus faber, and Trachurus mediterraneus. In contrast, the prevalence of Anisakis physeteris was only 1.3%. A hybrid genotype of Anisakis simplex sensu stricto and Anisakis pegreffii was also observed, which confirms the results of previous studies carried out in the western Mediterranean. Interestingly, no Anisakis simplex s.s. larvae were recovered. These results indicate that the diversity of Anisakis species is low in Sardinia waters, probably because of its geographic position.

Outbreak of Sciaenacotyle panceri (Monogenea) on cage-reared meagre Argyrosomus regius (Osteichthyes) from the western Mediterranean Sea

Two different batches of meagre Argyrosomus regius were stocked in the same floating cage located in north-eastern Sardinia (western Mediterranean Sea). After 4 mo, the fish started showing non-specific disease signs, such as lethargy, emaciation, gill anaemia and mortality. In total 65 specimens of meagre of both stocks were sampled for parasitological and microbiological analysis, and only 1 species of parasite, the microcotylid monogenean Sciaenacotyle panceri, was found on the gills of caged meagre. No protozoan or other metazoan parasites were found and no bacteria or viruses were isolated. After the outbreak, a total mortality rate of 5 to 10% was estimated for the older fish stock, while no noteworthy mortality was recorded in younger fish (<2%). This is the first account of S. panceri on the gills of cultured meagre as causative agent of disease and mortality. The effects of the presence of this parasite emphasise the need for disease control strategies of cultured meagre, and investigating the transfer of microcotylids from wild to caged fish.

Molecular characterization of Hysterothylacium aduncum (Nematoda: Raphidascaridae) from different fish caught off the Tunisian coast based on nuclear ribosomal DNA sequences

Larval forms of the genus Hysterothylacium have been previously reported in teleost fish from the North African coasts of central Mediterranean Sea by morphological analysis. In the present study, samples identified morphologically as Hysterothylacium aduncum (n = 62), from Merluccius merluccius, Trachurus mediterraneus and Pagellus erythrinus from different geographical locations of the Tunisian coasts, were genetically characterised by sequences of the first (ITS-1), the 5.8S and second (ITS-2) Internal Transcribed Spacers (ITS) of nuclear ribosomal DNA (rDNA). Comparison of the sequences obtained with those available in public gene databases confirmed that all the samples from the Tunisian coasts belong to a single species, namely H. aduncum. All specimens from the Tunisian coasts showed one indel in position 787 in ITS-2 sequences not reported by any of the previously published sequences from the Atlantic and Pacific Oceans, Adriatic Sea (Mediterranean Sea) and the East Greenland Sea, suggesting the existence of a population-specific pattern exhibiting a low differentiation of this parasite in this area. This is the first molecular characterization of H. aduncum from the Tunisian coasts using ITS rDNA sequences which allows the definition of genetic markers for their unequivocal identification, and provides further biological data on these nematodes in marine fish off the Tunisian coasts, improving the picture of the occurrence of these taxa in the North African coasts of central Mediterranean Sea.

Combined analysis of four mitochondrial regions allowed the detection of several matrilineal lineages of the lessepsian fish Fistularia commersonii in the Mediterranean Sea

The bluespotted cornetfish ( Fistularia commersonii ) is an Indo-Pacific species that in the last ten years colonized a large part of the Mediterranean basin. The aim of this study was to sequence some portions of the mitochondrial DNA (D-loop II, 16S, 12S and Cyt b) of this fish from different localities of the Mediterranean Sea, in order to evaluate the level of its genetic variability in this area. The genetic analysis performed on specimens from seven localities of Sardinia, Tunisia and Libya revealed the presence of at least five mitochondrial lineages. The results obtained, compared with previous studies, indicate that the use of a sufficient number of mitochondrial regions may allow a more accurate estimate of genetic variability in lessepsian invasions.

Redescriptions of Ligophoruscephali Rubtsova, Balbuena, Sarabeev, Blasco-Costa & Euzet, 2006 and L. chabaudi Euzet & Suriano, 1977 (Monogenea: Ancyrocephalidae), with notes on the functional morphology of the copulatory organ

Redescriptions of Ligophorus cephali Rubtsova, Balbuena, Sarabeev, Blasco-Costa & Euzet, 2006 and L. chabaudi Euzet & Suriano, 1977 based on original material from the Black Sea, the Mediterranean Sea and the Sea of Japan are presented. A comparison of samples of these two species from different regions was carried out with the aid of principal components analysis. The occurrence of L. chabaudi on Mugil cephalus in the Sea of Japan was confirmed. The functional morphology of the male copulatory organ was examined, and the use of the shape of this structure in the taxonomy of Ligophorus Euzet & Suriano, 1977 is discussed.