Molecular characterization of larval anisakid nematodes from marine fishes of Madeira by a PCR-based approach, with evidence for a new species

One-hundred and fifteen anisakid larvae from 3 different fish hosts, Aphanopus carbo, Scomber japonicus, and Trachurus picturatus, caught in Madeiran waters, were identified by PCR-RFLP. Three distinct species were identified in A. carbo, namely Anisakis simplex sensu srricto, Anisakis pegreffii, and Anisakis ziphidarum; 5 in S. japonicus, i.e., A. simplex s.s., A. pegreffii, Anisakis physeteris, Anisakis typica, and A. ziphidarum; and 3 in T. picturatus, i.e., A. simplex s.s., A. pegreffii, and A. typica. Anisakis simplex s.s. was the most frequent species in both A. carbo and S. japonicus (54% and 23.5%, respectively). Anisakis pegreffii and A. physeteris occurred with a frequency of 20.6% in S. japonicus, whereas in T. picturatus the most frequent species was A. typica (41.9%), followed by A. simplex s.s. (32.3%). Furthermore, A. carbo and S. japonicus were infected by an apparently undescribed taxon, provisionally named Anisakis sp. A. Based on estimations of the genetic distance, this new taxon seems to be more similar to A. ziphidarum (0.0335) than to other species of the genus.     

Larval trypanorhynchs (Platyhelminthes: Eucestoda: Trypanorhyncha) from black-scabbard fish, Aphanopus carbo and oceanic horse mackerel, Trachurus picturatus in Madeira (Portugal)

Four different types of trypanorhynch postlarvae were found attached to the stomach mucosa, external stomach wall or free in the body cavity of two marine fishes from Madeira, namely blackscabbard fish, Aphanopus carbo and oceanic horse mackerel, Trachurus picturatus. Morphological features shown by SEM indicated that the postlarvae belonged to the species Tentacularia coryphaenae, Sphyriocephalus tergestinus, Nybelinia lingualis and possibly N. yamaguitii. Prevalence [mean intensity (range)] of T. coryphaenae, S. tergestinus and Nybelinia spp. in A. carbo (n = 135) was 12.6% [1.65 +/- 1.27(1-6)], 5.9% [1.57 +/- 0.79 (1-3)] and 2.2% [1.33 +/- 0.58 (1-2)] respectively. The prevalence of T. coryphaenae and S. tergestinus showed some seasonality, with a rise in prevalence of T. coryphaenae corresponding to a decrease in prevalence of S. tergestinus. However these differences were not significant. In T. picturatus (n = 304) only N. lingualis was found at a prevalence of 9.6%. Both S. tergestinus and N. lingualis were recovered only from the stomach mucosa or external stomach wall, while T. coryphaenae was observed either attached to the stomach mucosa or free in the visceral cavity of the fish. The paper presents the first scanning electron micrographs (SEM) of Sphyriocephalus tergestinus and a new geographical record of N. lingualis in T. picturatus.     

Gill monogeneans of the chub mackerel, Scomber japonicus from Madeiran waters of the Atlantic Ocean, Portugal

Five species of monogeneans were recovered from the gill filaments of 181 chub mackerel, Scomber japonicus, from the Madeiran waters of the Atlantic Ocean, Portugal, during 2004/2005. The monogenean Pseudokuhnia minor showed the highest prevalence (98.68%) and a mean intensity of 28.23, followed by Kuhnia scombri (prevalence of 43.71% and mean intensity of 2.69) and K. scombercolias (prevalence of 39.1% and a mean intensity of 1.81). Kuhnia sprostonae and Grubea cochlear were rare, occurring in only one and five fish hosts respectively. No correlation between fish host length and mean intensity of infection with the three most abundant monogeneans was found. However, significant differences in prevalence and abundance were found in relation to date of sampling for P. minor, and all parasites were aggregated in their distribution.     

Helminth parasites of the oceanic horse mackerel Trachurus picturatus Bowdich 1825 (Pisces: Carangidae) from Madeira Island, Atlantic Ocean, Portugal

The helminth parasite fauna of the oceanic horse mackerel Trachurus picturatus Bowdich 1825, caught off the Madeira Islands was composed of six different taxa. Prevalence and abundance of larval Anisakis sp. (Nematoda: Anisakidae) and Nybelinia lingualis (Trypanorhyncha: Tentaculariidae), the most common parasite taxa, were 24.3%, 0.9 and 37.9%, 0.7, respectively. Bolbosoma vasculosum (Acanthocephala: Polymorphidae) and the monogeneans Heteraxinoides atlanticus (Monogenea: Heteraxinidae) and Pseudaxine trachuri (Monogenea: Gastrocotylidae) were comparatively rare. The depauperate helminth fauna of the oceanic horse mackerel at Madeira compared to other geographical regions of the north-eastern Atlantic, namely the Azores banks and the West African coast, may be attributed to the paucity of nutrients off oceanic islands and to a low density of the fish population.     

The parasite fauna of the chub mackerel (Scombridae: Scomber japonicus Houttuyn, 1782) in the central-eastern Atlantic (Atlantic coast of the Northern Africa and the Azores Archipelago banks)

The parasite fauna of the chub mackerel Scomber japonicus Houtuym, 1782 was studied from the neritic areas of Morocco, Western Sahara, Mauritania and from the banks of the Azores Archipelago (the Great Meteor Bank, the Hyeres Bank and the Irving Bank) in 1994-2001. Twenty eight species of parasites of following group have been were found: Coccidia (1 species), Microsporidia (1), Myxosporea (4), Monogenea (4), Cestoda (5), Trematoda (5), Acanthocephala (1) and Nematoda (6). The differences between mackerel parasite fauna in the neritic areas and from of the Azores Archipelago banks were established. Peculiarities of the mackerel parasite fauna in two areas (Morocco--Western Sahara and Mauritania) corroborate the hypothesis that two populations of chub mackerel are available: "Sahara-Moroccan" and "Senegal-Mauritanian". Ontogenetic variability of parasite fauna was related to food demands of mackerel and its feeding habits in the areas Morocco and Mauritania. Kudoa histolytica has negative influence on the commercial value of S. japonicus. These parasites were localized in the muscles of mackerel from Mauritania (40%, TL = 20-25 cm). Parasites being dangerous for human health were presented by larvae of Bolbosoma sp. (occurred on the banks of the Azores Archipelago), Anisakis simplex and Contracaecum sp. (occurred in all areas investigated).     

Anisakis simplex larvae: infection status in marine fish and cephalopods purchased from the Cooperative Fish Market in Busan, Korea

The infection status of marine fish and cephalopods with Anisakis simplex third stage larva (L3) was studied over a period of 1 year. A total of 2,537 specimens, which consisted of 40 species of fish and 3 species of cephalopods, were purchased from the Cooperative Fish Market in Busan, Korea, from August 2006 to July 2007. They were examined for A. simplex L3 from the whole body cavity, viscera, and muscles. A. simplex L3 were confirmed by light microscopy. The overall infection rate reached 34.3%, and average 17.1 larvae were parasitized per infected fish. Fish that recorded the highest infection rate was Lophiomus setigerus (100%), followed by Liparis tessellates (90%), Pleurogrammus azonus (90%), and Scomber japonicus (88.7%). The intensity of infection was the highest in Gadus macrocephalus (117.7 larvae per fish), followed by S. japonicus (103.9 larvae) and L. setigerus (54.2 larvae). Although abundance of A. simplex L3 was not seasonal in most of the fish species, 10 of the 16 selected species showed the highest abundance in February and April. A positive correlation between the intensity of L3 infection and the fish length was obvious in S. japonicus and G. macrocephalus. It was likely that A. simplex L3 are more frequently infected during the spring season in some species of fish. Our study revealed that eating raw or undercooked fish or cephalopods could still be a source of human infection with A. simplex L3 in Korea.     

Geographical variations in the jack mackerel Trachurus symmetricus murphyi populations in the southeastern Pacific Ocean as evidenced from the associated parasite communities

The composition of the metazoan parasite communities within 2 fishing zones along the Chilean coast were compared to determine the population structure of the jack mackerel Trachurus symmetricus murphyi (Nichols) in these 2 geographical locations. More than 7,780 parasites belonging to 15 taxa were collected between 1990 and 1996 in 71 samples comprised of 3,946 hosts. The same taxa were found in both fishing zones. However, jack mackerel from northern Chile had a higher abundance of cymothoid isopods, Ceratothoa spp., whereas those from southern Chile had more Rhadinorhynchus trachuri, Hysterothylacium sp. larvae, and Anisakis type I larvae. Results were similar at all community levels because all parasites had low prevalence and infracommunities had low total abundance and richness. Analyses at the component community level may be more adequate for stock identification when infracommunities are simple. There were significant differences in composition of parasite communities between years in each fishing zone, presumably as a result of the increase in offshore catches since 1994. These results reinforce the hypothesis that more than 1 ecological stock of the jack mackerel exists in the southeastern Pacific and contradict the current assumption of a single stock in the management of this heavily exploited fish species.     

Host and geographic range extensions of the North American strain of viral hemorrhagic septicemia virus

Viral hemorrhagic septicemia virus (VHSV) was isolated from populations of Pacific sardine Sardinops sagax from the coastal waters of Vancouver Island, British Columbia, Canada, and central and southern California, USA. The virus was also isolated from Pacific mackerel Scomber japonicus in southern California, from eulachon or smelt Thaleichthys pacificus, and surf smelt Hypomesus pretiosus pretiosus from Oregon, USA. Mortality and skin lesions typical of viral hemorrhagic septicemia in other marine fish species were observed among sardine in Canada and in a few surf smelt from Oregon, but the remaining isolates of VHSV were obtained from healthy appearing fish. The prevalence of VHSV among groups of apparently healthy sardine, mackerel and smelt ranged from 4 to 8%, in California and Oregon. A greater prevalence of infection (58%) occurred in groups of sardine sampled in Canada that sustained a naturally occurring epidemic during 1998-99. A captive group of surf smelt in Oregon exhibited an 81% prevalence of infection with clinical signs in only a few fish. The new isolates were confirmed as North American VHSV and were closely related based on comparisons of the partial nucleotide sequence of the glycoprotein (G) gene. The VHSV isolates from sardine in Canada and California were the most closely related, differing from isolates obtained from other marine fish species and salmonids in British Columbia, Canada, Alaska and Washington, USA. These new virus isolations extend both the known hosts (sardine, mackerel and 2 species of smelt) and geographic range (Oregon and California, USA) of VHSV.     

Differential population structuring of two closely related fish species, the mackerel (Scomber scombrus) and the chub mackerel (Scomber japonicus), in the Mediterranean Sea

Population genetic structures of the mackerel (Scomber scombrus) and chub mackerel (Scomber japonicus) were studied in the Mediterranean Sea. Fragments of 272 bp (S. scomber) and 387 bp (S. japonicus) of the 5'-end of the mitochondrial control region were sequenced from spawning individuals collected off the coasts of Greece, Italy, Spain, and Portugal. High levels of mitochondrial control region haplotypic diversity (> 0.98) were found for both Scomber species. Nucleotide diversity was higher in the mackerel (0.022) than in the chub mackerel (0.017). Global F(ST) values were also higher and significant in the mackerel (0.024, P < 0.0001) as opposed to the chub mackerel (0.003, P > 0.05). Molecular variance analyses showed differential genetic structuring for these two closely related species. There is extensive gene flow between Mediterranean Sea and Atlantic Ocean populations of chub mackerel, which are organized into a larger panmictic unit. In contrast, Mediterranean Sea populations of mackerel show some degree of genetic differentiation and are structured along an east-west axis. The analysed eastern Mediterranean Sea mackerel populations (Greece, Italy) are clearly separated from that of the western Mediterranean Sea (Barcelona), which forms a panmictic unit with eastern Atlantic Ocean populations. The genetic structures of both species showed asymmetric migration patterns and indicated population expansion.     

Metazoan parasites of the jack mackerel Trachurus murphyi (Teleostei, Carangidae) in a latitudinal gradient from South America (Chile and Peru).

The metazoa parasite fauna of the jack mackerel Trachurus murphyi Nichols 1920, obtained from seven ports from a latitudinal gradient of more than 30 degrees (From Paita Peru: 05 degrees 10' S to Talcahuano, Chile 36 degrees 40' S) was quantitatively described. 21 species of metazoan parasites were recorded (Monogenea 4, Digenea 3, Cestoda 5, Acanthocephala 2, Nematoda 4, Copepoda 2, Isopoda 1). Only four parasites (Lernanthropus trachuri, Ceratothoa gaudichaudii, Corynosoma australis, Anisakis simplex) were common to the seven ports. Only twelve species reach prevalence over 5% in at least one port. Main characteristic of the parasite fauna is the sparse of adult helminthes, specifically digenean and the dominance of larval forms, at both qualitative and quantitative levels. There are not evidence of changes in the parasite community along a latitudinal gradient, but at least Anisakis simplex (larvae) shows a trend in both, prevalence and mean intensity, increasing from north to south.     

Molecular and morphologic approaches to discrimination of variability patterns in chub mackerel, Scomber japonicus

The systematic status and the evolutionary biology of chub mackerel (Scomber japonicus) in the South West Atlantic Ocean is confusing with an unknown degree of genetic differentiation and reproductive isolation between units. Simultaneous genetic and morphologic analyses were made on 227 fish collected from two areas of the South West Atlantic Ocean and one from the Mediterranean Sea. The genetic analysis was based on 36 protein-coding loci, 16 of which were variable. The morphologic analyses include six morphometric length measurements and a meristic character. Correspondence between genetic and morphologic variability patterns indicates isolated Mediterranean and Southwest Atlantic subgroups of S. japonicus and, less clearly, possible additional divergence in two regional stocks within the latter group. The most conservative approach to management is to manage the stocks independently of one another.     

Variations in otolith patterns, sizes and body morphometrics of jack mackerel Trachurus japonicus juveniles

Variations in otolith patterns, sizes and body morphometrics of jack mackerel Trachurus japonicus juveniles were investigated. Under transmitted light, translucent (W(t)) and opaque otoliths (W(o)) were detected in juveniles collected from Wakasa Bay between July 2005 and April 2006, whereas only opaque otoliths (G(o)) were detected in Goto-nada Sea individuals between May and June 2006. Three groups of juveniles were distinguished based on differences in hatch season, otolith size and growth history, and body morphometrics. As T. japonicus has different spawning seasons according to spawning grounds, each group was estimated to hatch in different waters. Juveniles with W(t) otoliths were considered to have stayed in coastal habitat longer, as the hatch area was estimated to be near Wakasa Bay. Juveniles with W(o) and G(o) otoliths appear to recruit to coastal waters at larger size, since their hatch areas were estimated to be far from each collection area. Larger otoliths of W(t) were attributed to otolith accretion after the second growth flexion, which was observed only for W(t) . Standard length of W(t) fish at the second otolith growth flexion was estimated to correspond to recruitment size to coastal rocky reefs in Wakasa Bay. Body morphometrics were correlated with otolith size after removing body size effect, suggesting that morphological variations of T. japonicus juveniles were also associated with the timing of recruitment to coastal habitat.     

Ontogeny of swimming speed, schooling behaviour and jellyfish avoidance by Japanese anchovy Engraulis japonicus

The ontogeny of swimming speed, schooling behaviour and jellyfish avoidance was studied in hatchery-reared Japanese anchovy Engraulis japonicus to compare its life-history strategy with two other common pelagic fishes, jack mackerel Trachurus japonicus and chub mackerel Scomber japonicus. Cruise swimming speed of E. japonicus increased allometrically from 1·4 to 3·9 standard length (L(S) ) per s (L(S) s(-1) ) from early larval to metamorphosing stage. Burst swimming speed also increased from 6·1 to 28 L(S) s(-1) in these stages. Cruise speed was inferior to that of S. japonicus, as was burst speed to that of T. japonicus. Engraulis japonicus larvae were highly vulnerable to predation by moon jellyfish Aurelia aurita and were readily eaten until they reached 23 mm L(S) , but not at 26 mm L(S) . Schooling behaviour (indicated by parallel swimming) started at c. 17 mm L(S) . Average distance to the nearest neighbour was shorter than values reported in other pelagic fishes. The relatively low predator avoidance capability of E. japonicus may be compensated for by their transparent and thus less conspicuous body, in addition to their early maturation and high fecundity.     

The diet of blue whiting, hake, horse mackerel and mackerel off Portugal

This paper deals with the diets of blue whiting Micromesistius poutassou (Risso 1810), hake Merluccius merluccius (L. 1758), horse mackerel Trachurus trachurus (L. 1758), and mackerel Scomber scombrus (L. 1758) off Portugal and explores variations in fish length, water depth, latitude and season. All four species feed on fish; however, hake and mackerel are the first and second most important predators, respectively, blue whiting being the most important fish prey for both species. The diets of blue whiting and horse mackerel are composed mainly of crustaceans. Diet variations according to predator fish size are more important than either latitude or depth. In the diets of blue whiting, hake and horse mackerel, prey importance increases with predator size. For blue whiting and horse mackerel, diet variations with fish length and water depth are correlated: small fish are closely associated with coastal areas where they feed on copepods and decapod larvae. Seasonality in the diet is apparent for blue whiting, hake and mackerel. For blue whiting, the decapod Pasiphaea sivado is the most important prey in summer and autumn, being replaced by the euphausid Meganyctiphanes norvegica in winter. In the diet of hake, seasonality was characterised by the major importance of Macroramphosus scolopax in autumn, whereas the diet of mackerel consisted of zooplankton in summer, fish and decapods in autumn and decapod larvae in winter. Seasonal changes in the diet of horse mackerel correspond to a higher diversity of prey in autumn compared to other seasons (although euphausids are the main prey in all seasons). Seasonality in feeding activity is not as marked for the other species as it is for horse mackerel; the percentage of empty stomachs of horse mackerel is greatest in winter, when spawning takes place at the Portuguese coast.     

Anisakis spp. in fishery products from Japanese waters: Updated insights on host prevalence and human infection risk factors

The nematodes of the genus Anisakis are among the most relevant parasitic hazards in fishery products since they are responsible for human infection and allergy cases. In a food safety and epidemiological perspective, several marine hosts from different locations around Japan were examined to characterize the parasitism of Anisakis larvae. Chum salmon (Oncorhynchus keta) and Alaska pollock (Gadus chalcogrammus) showed the highest overall prevalence (100%), followed by blue mackerel (Scomber australasicus) (97.5%), Pacific cod (Gadus macrocephalus) (80%), chub mackerel (Scomber japonicus) (60.1%), Japanese flying squid (Todarodes pacificus) (17%) and Japanese pilchard (Sardinops sagax melanostictus) (2%). In Pacific krill (Euphausia pacifica), apart from one Hysterothylacium aduncum larva, no Anisakis specimens were detected. Anisakis simplex sensu stricto was molecularly identified (PCR-RFLP) for the first time in Japanese flying squid and Japanese pilchard distributed in the Northwestern Pacific ocean. That was the most frequent parasitic species detected followed by A. pegreffii, mostly in the western areas of Japan, hybrid genotypes between the two sibling species as well as A. typica and A. berlandi. Surprisingly, A. simplex s.s. was the most abundant species in one batch of chub mackerel from the East China Sea and A. pegreffii was the main species found in one batch from the Pacific coast of Aomori, which seems to indicate that the ranges of these two sibling species might be more variable than previously thought.     

PERMANENT GENETIC RESOURCES: Isolation and characterization of nine microsatellite loci from the chub mackerel, Scomber japonicus (Perciformes, Scombridae)

The stock abundance of the chub mackerel Scomber japonicus - a very important species for fisheries, particularly in Japan - in the Pacific Ocean off Japan has remained at a low level. For studying the population genetics of the chub mackerel, we isolated nine polymorphic microsatellite loci (12-31 alleles/locus; expected heterozygosity, 0.762-0.983) from this species. Cross-species amplification indicated that eight of the nine microsatellite loci in the blue mackerel S. australasicus were polymorphic and functional.     

Purification and cloning of an apoptosis-inducing protein derived from fish infected with Anisakis simplex, a causative nematode of human anisakiasis

While investigating the effect of marine products on cell growth, we found that visceral extracts of Chub mackerel, an ocean fish, had a powerful and dose-dependent apoptosis-inducing effect on a variety of mammalian tumor cells. This activity was strikingly dependent on infection of the C. mackerel with the larval nematode, Anisakis simplex. After purification of the protein responsible for the apoptosis-inducing activity, we cloned the corresponding gene and found it to be a flavoprotein. This protein, termed apoptosis-inducing protein (AIP), was also found to possess an endoplasmic reticulum retention signal (C-terminal KDEL sequence) and H2O2-producing activity, indicating that we had isolated a novel reticuloplasimin with potent apoptosis-inducing activity. AIP was induced in fish only after infection with larval nematode and was localized to capsules that formed around larvae to prevent their migration to host tissues. Our results suggest that AIP may function to impede nematode infection.     

Molecular characterization of Anisakis larvae from fish caught off Sardinia

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.     

Diet and trophic position of two mackerel species in the archipelago of Madeira,Portugal

The Atlantic chub mackerel Scomber colias and the blue jack mackerel Trachurus picturatus are two abundant species in the Macaronesia region which includes the archipelago of Madeira, Portugal. Both are key species in the trophic web, being important prey for several local top predators, such as seabirds and marine mammals. Nonetheless, little is known about their feeding ecology in oceanic environments. In this study, the authors describe the seasonal variation in the diet of S. colias and T. picturatus in the oceanic region of Madeira throughout a year. Visual inspection of stomach contents revealed that S. colias fed on a broader range of prey groups than T. picturatus, but for both species, zooplankton (particularly calanoid copepods) and fish were the most important food items. The diet of S. colias included a higher proportion of fish, namely Atlantic saury Scomberesox saurus and S. colias, than that of T. picturatus, that included mostly the longspine snipefish Macroramphosus scolopax. T. picturatus consumed a higher proportion of decapods and other copepods. Seasonal variation was found in the diet of both species, with zooplanktonic species being more important in colder months (February to April) for S. colias and during warm months (May to October) for T. picturatus. Their diet in other seasons was dominated by fish. Although they consume similar prey, carbon and nitrogen stable isotope analysis of muscle of S. colias and T. picturatus showed little overlap in their diets, and T. picturatus showed higher δ¹⁵N and a narrower isotopic niche.