Reliably discriminating stock structure with genetic markers: Mixture models with robust and fast computation.

Delineating naturally occurring and self-sustaining subpopulations (stocks) of a species is an important task, especially for species harvested from the wild. Despite its central importance to natural resource management, analytical methods used to delineate stocks are often, and increasingly, borrowed from superficially similar analytical tasks in human genetics even though models specifically for stock identification have been previously developed. Unfortunately, the analytical tasks in resource management and human genetics are not identical-questions about humans are typically aimed at inferring ancestry (often referred to as "admixture") rather than breeding stocks. In this article, we argue, and show through simulation experiments and an analysis of yellowfin tuna data, that ancestral analysis methods are not always appropriate for stock delineation. In this work, we advocate a variant of a previously introduced and simpler model that identifies stocks directly. We also highlight that the computational aspects of the analysis, irrespective of the model, are difficult. We introduce some alternative computational methods and quantitatively compare these methods to each other and to established methods. We also present a method for quantifying uncertainty in model parameters and in assignment probabilities. In doing so, we demonstrate that point estimates can be misleading. One of the computational strategies presented here, based on an expectation-maximization algorithm with judiciously chosen starting values, is robust and has a modest computational cost.

Species identification in mixed tuna samples with next-generation sequencing targeting two short cytochrome b gene fragments.

Conventional Sanger sequencing of PCR products is the gold standard for species authentication of seafood products. However, this method is inappropriate for the analysis of products that might contain mixtures of species, such as tinned tuna. The purpose of this study was to test whether next-generation sequencing (NGS) can be a solution for the authentication of mixed products. Nine tuna samples containing mixtures of up to four species were prepared and subjected to an NGS approach targeting two short cytochrome b gene (cytb) fragments on the Illumina MiSeq platform. Sequence recovery was precise and admixtures of as low as 1% could be identified, depending on the species composition of the mixtures. Duplicate samples as well as two individual NGS runs produced very similar results. A first test of three commercial tinned tuna samples indicated the presence of different species in the same tin, although this is forbidden by EU law.

Bycatch in the Maldivian pole-and-line tuna fishery.

Tropical tuna fisheries are among the largest worldwide, with some having significant bycatch issues. However, pole-and-line tuna fisheries are widely believed to have low bycatch rates, although these have rarely been quantified. The Maldives has an important pole-and-line fishery, targeting skipjack tuna (Katsuwonus pelamis). In the Maldives, 106 pole-and-line tuna fishing days were observed between August 2014 and November 2015. During 161 fishing events, tuna catches amounted to 147 t: 72% by weight was skipjack, 25% yellowfin tuna (Thunnus albacares) and 3% other tunas. Bycatch (all non-tuna species caught plus all tuna discards) amounted to 951 kg (0.65% of total tuna catch). Most of the bycatch (95%) was utilized, and some bycatch was released alive, so dead discards were particularly low (0.02% of total tuna catch, or 22 kg per 100 t). Rainbow runner (Elagatis bipinnulata) and dolphinfish (Coryphaena hippurus) together constituted 93% of the bycatch. Live releases included small numbers of silky sharks (Carcharhinus falciformis) and seabirds (noddies, Anous tenuirostris and A. stolidus). Pole-and-line tuna fishing was conducted on free schools and schools associated with various objects (Maldivian anchored fish aggregating devices [aFADs], drifting FADs from western Indian Ocean purse seine fisheries, other drifting objects and seamounts). Free school catches typically included a high proportion of large skipjack and significantly less bycatch. Associated schools produced more variable tuna catches and higher bycatch rates. Fishing trips in the south had significantly lower bycatch rates than those in the north. This study is the first to quantify bycatch rates in the Maldives pole-and-line tuna fishery and the influence of school association on catch composition. Ratio estimator methods suggest roughly 552.6 t of bycatch and 27.9 t of discards are caught annually in the fishery (based on 2015 national catch), much less than other Indian Ocean tuna fisheries, e.g. gillnet, purse-seine, and longline.

Incidence of three Kudoa spp., K. neothunni, K. hexapunctata, and K. thunni (Myxosporea: Multivalvulida), in Thunnus tunas distributed in the western Pacific Ocean.

A variety of tunas of the genus Thunnus are consumed daily in Japan as sliced raw fish (sashimi and sushi). The consumption of fresh sliced raw fish, i.e., unfrozen or uncooked, can sometimes cause food poisoning that is manifested by transient diarrhea and vomiting for a single day. One of the causes of this type of food poisoning has been identified as live Kudoa septempunctata (Myxosporea: Multivalvulida) in the olive flounder (Paralichthys olivaceus). Furthermore, raw slices of fresh tunas are highly suspected to be a possible causative fish of similar food poisoning in Japan. In the present study, we conducted a survey of kudoid infections in tunas (the yellowfin tuna Thunnus albacares, the Pacific bluefin tuna Thunnus orientalis, and the longtail tuna Thunnus tonggol) fished in the western Pacific Ocean off Japan and several East Asian countries and characterized morphologically and genetically the kudoid myxospores in pseudocysts or cysts dispersed in the trunk muscles. Pseudocysts of solely Kudoa hexapunctata were identified in the Pacific bluefin tuna (four isolates), whereas in the yellowfin tuna (21 isolates) pseudocysts of Kudoa neothunni and K. hexapunctata were detected at a ratio of 15:6, respectively, in addition to cyst-forming Kudoa thunni in five yellowfin tunas. In the trunk muscles of six longtail tunas examined, pseudocysts of K. neothunni (all six fish) and K. hexapunctata (two fish) were densely dispersed. The myxospores of K. neothunni found in these longtail tunas had seven shell valves and polar capsules (SV/PC) instead of the more common six SV/PC arranged symmetrically. Nucleotide sequences of the 18S and 28S ribosomal RNA gene (rDNA), some with the internal transcribed spacer regions as well, of K. hexapunctata and K. neothunni from the three Thunnus spp., including the seven-SV/PC morphotype, were very similar to previously characterized nucleotide sequences of each species, whereas the 18S and 28S rDNA of four isolates of K. thunni from yellowfin tunas showed a range of nucleotide variations of 99.0-99.9% identity over 1752-1763-bp long partial 18S rDNA and 97.4-99.9% identity over 797-802-bp long partial 28S rDNA. Therefore, this rather high variation of the rDNA nucleotide sequences of K. thunni proved to be contrary to the few variations of K. neothunni and K. hexapunctata rDNA nucleotide sequences. The present study provides a new host record of the longtail tuna for K. neothunni and K. hexapunctata and reveals a high prevalence of the seven-SV/PC myxospore morphotype of K. neothunni in this tuna host.

Development of a qPCR Method for the Identification and Quantification of Two Closely Related Tuna Species, Bigeye Tuna (Thunnus obesus) and Yellowfin Tuna (Thunnus albacares), in Canned Tuna.

Bigeye tuna (Thunnus obesus) and yellowfin tuna (Thunnus albacares) are among the most widely used tuna species for canning purposes. Not only substitution but also mixing of tuna species is prohibited by the European regulation for canned tuna products. However, as juveniles of bigeye and yellowfin tunas are very difficult to distinguish, unintentional substitutions may occur during the canning process. In this study, two mitochondrial markers from NADH dehydrogenase subunit 2 and cytochrome c oxidase subunit II genes were used to identify bigeye tuna and yellowfin tuna, respectively, utilizing TaqMan qPCR methodology. Two different qPCR-based methods were developed to quantify the percentage of flesh of each species used for can processing. The first one was based on absolute quantification using standard curves realized with these two markers; the second one was founded on relative quantification with the universal 12S rRNA gene as the endogenous gene. On the basis of our results, we conclude that our methodology could be applied to authenticate these two closely related tuna species when used in a binary mix in tuna cans.

RAD-seq derived genome-wide nuclear markers resolve the phylogeny of tunas.

Although species from the genus Thunnus include some of the most commercially important and most severely overexploited fishes, the phylogeny of this genus is still unresolved, hampering evolutionary and traceability studies that could help improve conservation and management strategies for these species. Previous attempts based on mitochondrial and nuclear markers were unsuccessful in inferring a congruent and reliable phylogeny, probably due to mitochondrial introgression events and lack of enough phylogenetically informative markers. Here we infer the first genome-wide nuclear marker-based phylogeny of tunas using restriction site associated DNA sequencing (RAD-seq) data. Our results, derived from phylogenomic inferences obtained from 128 nucleotide matrices constructed using alternative data assembly procedures, support a single Thunnus evolutionary history that challenges previous assumptions based on morphological and molecular data.

The differentiation of tuna (family: Scombridae) products through the PCR-based analysis of the cytochrome b gene and parvalbumin introns.

BACKGROUND: In spite of the many studies performed over the years, there are still problems in the authentication of closely related tuna species, not only for canned fish but also for raw products. With the aim of providing screening methods to identify different tuna species and related scombrids, segments of mitochondrial cytochrome b (cyt b) and nuclear parvalbumin genes were amplified and sequenced or subjected to single-strand conformation polymorphism (SSCP) and restriction fragment length polymorphism (RFLP) analyses. RESULTS: The nucleotide diagnostic sites in the cyt b gene of five tuna species from Indonesia were determined in this study and used to construct a phylogenetic tree. In addition, the suitability of the nuclear gene that encodes parvalbumin for the differentiation of tuna species was determined by SSCP and RFLP analyses of an intron segment. RFLP differentiated Thunnus albacares and from T. obesus, and fish species in the Thunnus genus could be distinguished from bullet tuna (Auxis rochei) by SSCP. CONCLUSIONS: Parvalbumin-based polymerase chain reaction systems could serve as an additional tool in the detection and identification of tuna and other Scombridae fish species for routine seafood control. This reaction can be performed in addition to the cyt b analysis as previously described.

The sequence and organization of complete mitochondrial genome of the yellowfin tuna, Thunnus albacares (Bonnaterre, 1788).

Yellowfin tuna (Thunnus albacares) is one of the most important economic fishes around the world. In the present study, we determined the complete mitochondrial DNA sequence and organization of T. albacares. The entire mitochondrial genome is a circular-molecule of 16,528 bp in length, which encodes 37 genes in all. These genes comprise 13 protein-coding genes (ATP6 and 8, COI-III, Cytb, ND1-6 and 4 L), 22 transfer RNA genes (tRNAs), and 2 ribosomal RNA genes (12S and 16S rRNAs). The complete mitochondrial genome sequence of T. albacares can provide basic information for the studies on molecular taxonomy and conservation genetics of teleost fishes.

Identification of five highly priced tuna species by quantitative real-time polymerase chain reaction.

Tunas are economically important fishery worldwide, and are often used for commercial processed production. For effective fishery management and protection of consumers' rights, it is important to develop a molecular method to identify species in canned tuna products rapidly and reliably. Here, we have developed a duplex quantitative real-time PCR (qPCR) for identification of five highly priced tuna species (Thunnus maccoyii, Thunnus obesus, Thunnus albacares, Thunnus alalunga and Katsuwonus pelamis) from processed as well as fresh fish. After amplification and sequencing of seven genetic markers commonly used for species identification, 16S rDNA and control region (CR) of mitochondrial DNA were selected as the reference gene markers for genus Thunnus and tuna species identification, respectively. Subsequently, a 73 bp fragment of 16S rDNA and 85-99 bp fragment of CR were simultaneously amplified from each target species by qPCR. The qPCR efficiency of each reaction was calculated according to the standard curves, and the method was validated by amplification DNA extracted from single or mixed tuna specimen. The developed duplex qPCR system was applied to authenticate species of 14 commercial tuna products successfully, which demonstrated it was really a useful and academic technique to identify highly priced tuna species.

Mitochondrial genome of the blackfin tuna Thunnus atlanticus Lesson, 1831 (Perciformes, Scrombidae).

Blackfin tuna, Thunnus atlanticus is a widespread epipelagic oceanic species in the western Atlantic. So far the mitochondrial genome of this species remained unknown, although the mitogenomes of all congeners are known. The mitochondrial genome encodes for 13 proteins, 21 tRNAs, 2 ribosomal RNAs and the gene synteny is conserved with other previously reported mitogenomes of tunas.

Mitochondrial genome of the bullet tuna Auxis rochei from Indo-West Pacific collection provides novel genetic information about two subspecies.

Previously morphological studies supported the division of the bullet tuna into the two subspecies, Auxis rochei rochei and A. rochei eudorax. As a cosmopolitan species, A. rochei rochei ranges in the Indo-West Pacific and Atlantic oceans, while A. rochei eudorax inhabits in eastern Pacific region. Here, we used the HiSeq next-generation sequencing technique to determine the mitochondrial genome (mitogenome) of A. rochei from Indo-West Pacific collection, and then compared our data with mitogenomic sequences of the Atlantic and eastern Pacific retrieved from NCBI database. Results showed the mitogenome of A. rochei from three geographic collections shared the same genes and gene order, similar to typical teleosts. Also, we examined a low level of nucleotide diversity among these mitogenomic sequences. Interestingly, nucleotide diversity of intra-subspecies (Atlantic versus Indo-West) was higher than that of inter-subspecies (Atlantic versus eastern Pacific, Indo-West versus eastern Pacific).

Evidence of discrete yellowfin tuna (Thunnus albacares) populations demands rethink of management for this globally important resource.

Tropical tuna fisheries are central to food security and economic development of many regions of the world. Contemporary population assessment and management generally assume these fisheries exploit a single mixed spawning population, within ocean basins. To date population genetics has lacked the required power to conclusively test this assumption. Here we demonstrate heterogeneous population structure among yellowfin tuna sampled at three locations across the Pacific Ocean (western, central, and eastern) via analysis of double digest restriction-site associated DNA using Next Generation Sequencing technology. The differences among locations are such that individuals sampled from one of the three regions examined can be assigned with close to 100% accuracy demonstrating the power of this approach for providing practical markers for fishery independent verification of catch provenance in a way not achieved by previous techniques. Given these results, an extended pan-tropical survey of yellowfin tuna using this approach will not only help combat the largest threat to sustainable fisheries (i.e. illegal, unreported, and unregulated fishing) but will also provide a basis to transform current monitoring, assessment, and management approaches for this globally significant species.

Assessing niche width of endothermic fish from genes to ecosystem.

Endothermy in vertebrates has been postulated to confer physiological and ecological advantages. In endothermic fish, niche expansion into cooler waters is correlated with specific physiological traits and is hypothesized to lead to greater foraging success and increased fitness. Using the seasonal co-occurrence of three tuna species in the eastern Pacific Ocean as a model system, we used cardiac gene expression data (as a proxy for thermal tolerance to low temperatures), archival tag data, and diet analyses to examine the vertical niche expansion hypothesis for endothermy in situ. Yellowfin, albacore, and Pacific bluefin tuna (PBFT) in the California Current system used more surface, mesopelagic, and deep waters, respectively. Expression of cardiac genes for calcium cycling increased in PBFT and coincided with broader vertical and thermal niche utilization. However, the PBFT diet was less diverse and focused on energy-rich forage fishes but did not show the greatest energy gains. Ecosystem-based management strategies for tunas should thus consider species-specific differences in physiology and foraging specialization.

Factors that determine the quality of bigeye tuna, caught in the western tropical Atlantic Ocean / Fatores que determinam a qualidade da albacora-bandolim, capturada no Oceano Atlântico oeste tropical

Tuna are highly prized in Oriental cuisine, and the bigeye tuna (Thunnus obesus) is of great commercial importance. Quality is assessed by the tuna meat quality, freshness, texture and fat, which can be altered by factors inherent to capture, such as days of onboard storage, months of the year, whether the catch was living or dead, fish weight and year. These factors were correlated to identify those that affect quality. We obtained data from nine vessels, from January 2007 to April 2010, creating an information bank of 21,908 bigeye tuna. Fish quality and parameters were related using multiple linear regression analysis, in which the variables were included in the model by a stepwise procedure (F>4). We found that live catch, heavier fish and fewer days of storage positively affected the quality, and that there is a seasonality of quality related to biological factors, which are not amenable to control...

Os atuns são peixes muito apreciados na culinária oriental, e a albacora-bandolim (Thunnus obesus) é de grande importância comercial. A qualidade do atum é avaliada pela qualidade da carne, frescor, textura e gordura, a qual pode ser alterada por fatores inerentes à captura, tais como: dias de armazenamento a bordo, meses do ano, capturado vivo ou morto, peso e ano. Esses fatores foram correlacionados para se identificarem aqueles que afetam a qualidade da carne. Foram obtidos dados de nove embarcações, de janeiro de 2007 a abril de 2010, e criou-se um banco de informações de 21.908 atuns. A qualidade do pescado e os parâmetros foram relacionados utilizando-se análise de regressão linear múltipla, em que as variáveis foram incluídas no modelo de stepwise (F> 4). Descobriu-se que captura do animal vivo, peixe mais pesado e menos dias de armazenamento afetaram positivamente a qualidade da carne e que há uma sazonalidade da qualidade relacionada a fatores biológicos que não são passíveis de controle...

Muscular system in the pacific bluefin tuna Thunnus orientalis (Teleostei: Scombridae).

The muscular system in the Pacific bluefin tuna Thunnus orientalis is studied in detail. For the first time, a complete description of the muscular anatomy of a thunnid is provided here. Eighty-two elements including subdivisions of components of the muscular system are identified. This is less than found in a basal perciform and two other investigated scombrid species, owing mainly to the absence or fusion of pectoral, pelvic and caudal fin muscles. The absence of elements of the basal perciform pattern was most prominent in the caudal fin, which includes only the flexor dorsalis, flexor ventralis, hypochordal longitudinalis, and interradialis. In the caudal fin, the medial fan-shaped ray was identified as the first dorsal ray, judging from myological and neuroanatomical characters. The highly developed gill filament muscles in Thunnus orientalis and sheet-like rectus communis control gill ventilation. Long body muscle tendons reduce the metabolic energy needed during rapid and continuous swimming. These characters are interpreted as adaptations in the context of the oceanic life style of the species.

Evolutionary history and phylogenetic relationship between Auxis thazard and Auxis rochei inferred from COI sequences of mtDNA.

Tunas of the genus Auxis are cosmopolitan species and the smallest members of the tribe Thunnini, the true tunas. In the present study, COI sequences of mtDNA were employed to examine the evolutionary history and phylogenetic relationship between A. thazard and A. rochei. A total of 29 COI sequences were retrieved from NCBI. Historic demographic analyses of sequence data showed that A. thazard has undergone sudden population expansion in the past while population size of A. rochei has been remain constant for long period. Non-significant value of Tajimas's D (P = 0.22400) and Fu's FS (P = 0.21400) test fail to reject the null hypothesis of neutral evolution for A. rochei. Phylogenetic analyses of nucleotide sequences demonstrated separate clusters for both species and are strongly supported by 98% bootstrap value. The results of the present study suggest the recent founding of A. thazard in world ocean while A. rochei represents the ancestral species.

Reproduction of Blackfin tuna Thunnus atlanticus (Perciformes: Scombridae) in Saint Peter and Saint Paul Archipelago, Equatorial Atlantic, Brazil.

The reproduction of Blackfin tuna Thunnus atlanticus has been described for coastal regions, and for a long time, this species was considered to be a strictly continental spawner. Recently, this species was observed around a seamount habitat 500 nautical miles Northeast of Brazil, located between South America and Africa. In this study we describe the reproductive biology of Blackfin tuna at Saint Peter and Saint Paul Archipelago (SPSPA). Male and female gonads were sampled from December 2008 to July 2010, and analyzed macro and microscopically. A total of 361 fish were sampled (247 males and 114 females). Males were more common than females, with a sex ratio of 2.2 male:1 female. The fork length (FL) of all sampled specimens ranged from 38 to 98cm, and larger length classes were more frequent in males. It was possible to distinguish six maturity phases for females: immature, developing, spawning capable, actively spawning, regressing and recovering. Five phases were identified for males: immature, developing, spawning capable, actively spawning and recovering. The gonad index (GI) mean monthly values ranged from 6.6 (SD = 4.1) to 58.4 (SD = 34.7) for females, and from 2.6 (SD = 1.3) to 66.2 (SD = 30.4) for males. For both sexes, the largest GI values were observed at the beginning of the first semester of the year. Size at first maturity was estimated at 48cm FL and 55cm FL for females and males respectively. Approximately 80% of the specimens were adults and considered to be in reproductive conditions. Histological analysis of the ovaries and testes showed that most of the specimens were sexually mature and were reproductively active during all months of the year. However, females with mature ovaries, with large amounts of hydrated oocytes and post-ovulatory follicles, were mainly found from December to March, thus these months may constitute the main spawning season in SPSPA. Batch fecundity varied between 272025 and 1,140584 oocytes for 56 and 68 cm FL females respectively. Oocyte development and spawning patterns suggest a multiple spawning behavior. The results revealed that Blackfin tuna is using the SPSPA as a spawning ground, similar to other species commonly observed in the same area during the same reproductive season.

Reproduction of Blackfin tuna Thunnus atlanticus (Perciformes: Scombridae) in Saint Peter and Saint Paul Archipelago, Equatorial Atlantic, Brazil / Reproducción del atún de aleta negra Thunnus atlanticus (Perciformes: Scombridae) en el Archipiélago San Pedro y San Pablo, Atlántico Ecuatorial, Brasil

The reproduction of Blackfin tuna Thunnus atlanticus has been described for coastal regions, and for a long time, this species was considered to be a strictly continental spawner. Recently, this species was observed around a seamount habitat 500 nautical miles Northeast of Brazil, located between South America and Africa. In this study we describe the reproductive biology of Blackfin tuna at Saint Peter and Saint Paul Archipelago (SPSPA). Male and female gonads were sampled from December 2008 to July 2010, and analyzed macro and microscopically. A total of 361 fish were sampled (247 males and 114 females). Males were more common than females, with a sex ratio of 2.2 ♂:1♀. The fork length (FL) of all sampled specimens ranged from 38 to 98cm, and larger length classes were more frequent in males. It was possible to distinguish six maturity phases for females: immature, developing, spawning capable, actively spawning, regressing and recovering. Five phases were identified for males: immature, developing, spawning capable, actively spawning and recovering. The gonad index (GI) mean monthly values ranged from 6.6 (SD=4.1) to 58.4 (SD=34.7) for females, and from 2.6 (SD=1.3) to 66.2 (SD=30.4) for males. For both sexes, the largest GI values were observed at the beginning of the first semester of the year. Size at first maturity was estimated at 48cm FL and 55cm FL for females and males respectively. Approximately 80% of the specimens were adults and considered to be in reproductive conditions. Histological analysis of the ovaries and testes showed that most of the specimens were sexually mature and were reproductively active during all months of the year. However, females with mature ovaries, with large amounts of hydrated oocytes and post-ovulatory follicles, were mainly found from December to March, thus these months may constitute the main spawning season in SPSPA. Batch fecundity varied between 272 025 and 1 140 584 oocytes for 56 and 68cm FL females respectively. Oocyte development and spawning patterns suggest a multiple spawning behavior. The results revealed that Blackfin tuna is using the SPSPA as a spawning ground, similar to other species commonly observed in the same area during the same reproductive season.

A pesar de la importancia de las capturas del atún de aleta negra Thunnus atlanticus en el noreste de Brasil y en las cercanías del Archipiélago San Pedro y San Pablo (ASPSP), hay una escasez de información sobre su reproducción, especialmente en las islas. La reproducción del atún de aleta negra en este archipiélago se estudió de diciembre 2008 a julio 2010. Con este fin, se recolectaron 361 gónadas de hembras y machos, los machos fueron predominantemente más frecuentes en la muestra total, con una proporción sexual de 1♀:2.2 ♂. La longitud furcal todos los ejemplares muestreados varió entre 38 y 98 cm, y los machos fueron más abundantes en las clases de mayor longitud. Los valores medios mensuales del índice gonadal (IG) variaron de 6.6 (SD=4.1) a 58.4 (SD=34.7) y de 2.6 (SD=1.3) a 66.2 (SD=30.4) para hembras y machos, respectivamente. Los mayores valores de IG para ambos sexos fueron observados el inicio del primer semestre. Los análisis histológicos mostraron que la mayoría de los ovarios y testículos correspondieron a especímenes aptos para reproducirse. Sin embargo, hembras con ovarios maduros, con gran cantidad de oócitos hidratados y folículos post ovulatorios, se encontraron principalmente de diciembre a marzo, que se considera es la época de reproducción del atún de aleta negra alrededor del ASPSP.

Molecular characterization of kudoid parasites (Myxozoa: Multivalvulida) from somatic muscles of Pacific bluefin (Thunnus orientalis) and yellowfin (T. albacores) tuna.

The public health importance of Kudoa infection in fish remains unclear. Recently in Japan a Kudoa species, K. septempunctata, was newly implicated as a causative agent of unidentified food poisoning related to the consumption of raw olive flounder. Other marine fishery products are also suspected as causative raw foods of unidentified food poisoning. For this study, we detected kudoid parasites from sliced raw muscle tissues of a young Pacific bluefin and an adult yellowfin tuna. No cyst or pseudocyst was evident in muscles macroscopically, but pseudocysts were detected in both samples histologically. One substitution (within 1100 bp overlap) and ten substitutions (within 753 bp overlap) were found respectively between the partial sequences of 18S and 28S rDNAs from both isolates. Nucleotide sequence similarity searching of 18S and 28S rDNAs from both isolates showed the highest identity with those of K. neothunni from tuna. Based on the spore morphology, the mode of parasitism, and the nucleotide sequence similarity, these isolates from a Pacific bluefin and a yellowfin tuna were identified as K. neothunni. Phylogenetic analysis of the 28S rDNA sequence revealed that K. neothunni is classifiable into two genotypes: one from Pacific bluefin and the other from yellowfin tuna. Recently, an unidentified kudoid parasite morphologically and genetically similar K. neothunni were detected from stocked tuna samples in unidentified food poisoning cases in Japan. The possibility exists that K. neothunni, especially from the Pacific bluefin tuna, causes food poisoning, as does K. septempunctata.

Cytochrome Oxidase I (COI) sequence conservation and variation patterns in the yellowfin and longtail tunas.

Tunas are commercially important fishery worldwide. There are at least 13 species of tuna belonging to three genera, out of which genus Thunnus has maximum eight species. On the basis of their availability, they can be characterised as oceanic such as Thunnus albacares (yellowfin tuna) or coastal such as Thunnus tonggol (longtail tuna). Although these two are different species, morphological differentiation can only be seen in mature individuals, hence misidentification may result in erroneous data set, which ultimately affect conservation strategies. The mitochondrial DNA cytochrome oxidase c subunit 1 (COI) gene is one of the most popular markers for population genetic and phylogeographic studies across the animal kingdom. The present study aims to study the sequence conservation and variation in mitochondrial Cytochrome Oxidase I (COI) between these two species of tuna. COI sequence analysis of yellowfin and longtail revealed the close relationship between them in Thunnus genera. The present study is the first direct comparison of mitochondrial COI sequences of these two tuna species.