Order Pleuronectiformes Research Articles

The complete mitochondrial genome sequence of Rikuzen flounder, Dexistes rikuzenius (Jordan & Starks, 1904)

In this study, the complete mitochondrial genome of the Rikuzen flounder (Dexistes rikuzenius) using next-generation sequencing and bioinformatics analysis is presented. The mitogenome was found to be a circular molecule 17,494 bp in length, containing 13 protein-coding genes (PCG), 22 transfer RNA (tRNA) genes, two ribosomal RNA (rRNA) genes, and one control region (D-loop). Proportional nucleotide composition was A: 28.80% (5,039 bp), T: 26.51% (4,638 bp), C: 28.72% (5,025 bp) and G: 15.96% (2,792 bp), with a combined A+T content of 55.32%. The phylogenetic relationships of the taxon with 19 other flatfish species in the order Pleuronectiformes were then analyzed based on the 13 PCGs. Dexistes rikuzenius was most closely clustered with Cleisthenes herzensteini within this order, suggesting a close relationship.

Rapid collagen peptide mass fingerprinting as a tool to authenticate Pleuronectiformes in the food industry

Reliable species identification of specimens in the food chain is crucial for detecting mislabelling fraud. While visual identification, DNA-analyses, and proteomic approaches have been used in the past, this research provides an alternative proteomic technique that has been trialled here to identify tissue samples from species within the order Pleuronectiformes (flatfish). Previously, archaeological research has used peptide mass fingerprinting to identify fish remains to species by extracting collagen Type I preserved in bones. As this type of collagen can also be found in the fins, skin and muscle of fishes (types of tissue that can be found throughout the food chain), collagen peptide mass fingerprinting has been applied here to identify modern fish specimens to species. In this study, MALDI-ToF MS peptide fingerprints are obtained from enzymatic digestion of collagen Type I from fin (or skin if fin was not available) and muscle tissues from six specimens of Pleuronectiformes. Using the already available diagnostic collagen peptide biomarkers for flatfish, all six specimens could be identified to the correct species for each of the tissue types. We recommend using this rapid approach for future screenings of modern flatfish in the food chain to detect mislabelling fraud, and also in more general applications of modern ecological studies of this order. It is likely that the methods can be applied to other Actinopterygian fishes.

Seafood mislabeling in Greek market using DNA barcoding

The mislabeling of seafood products is a worldwide observed issue even though labeling regulations have been established (either at local or European/International level). Various molecular methods have been developed for fish species identification and detection of fraud, with DNA barcoding being the most popular and accurate one. Here, we present the first large-scale analysis aimed at assessing the fish mislabeling rate in Greece, by amplifying a fragment of approximately 655bp of the COI gene in 285 fish products collected from multiple markets in Greece through a monitoring program of the Hellenic Food Authority (EFET) over a time span of 4 years. This study initiated in 2015 under a European Commission Recommendation on a coordinated control plan with a view to establishing the prevalence of fraudulent practices in the marketing of certain foods and was further supported by EFET for another 3 years in order to monitor the Greek market. Sequencing and species attribution was successful in 92.3% of the samples. In 12.9% of these (34 cases), discrepancies were detected between the declared and the identified species and therefore the products were characterized as mislabeled. This rate is low, compared to other studies in Greece and other regions of the Mediterranean Sea (e.g. Italy) and worldwide (e.g. Canada). On the other hand, it remains higher than the average global substitution rate of 8%, reported in a recent meta-analysis and the European rate of 6% detected during the EU coordinated control program in 2015. Flatfishes (order Pleuronectiformes) presented the highest mislabeling rate (28.3%) followed by gadiforms (order Gadiformes) with 13.7%. Although, in some cases the substitution can be unintentional due to similar morphological characteristics and geographical distribution among species, in 55.7% of the mislabeled samples the substituted species is of lower quality resulting inevitably also to an economic profit. Our results support the need for continuous and well-documented monitoring of Greek and European markets.

Temporal expression profiles of leptin and its receptor genes during early development and ovarian maturation of Cynoglossus semilaevis.

Leptin (Lep) plays a key role in the regulation of food intake and energy homeostasis in vertebrates. Our previous studies have provided evidence for the existence of two leptin genes (lepa and lepb) and one leptin receptor (lepr) gene in a flatfish, the half-smooth tongue sole (Cynoglossus semilaevis). However, the spatial-temporal expression patterns and possible roles of the leptin system during early development and ovarian maturation are still poorly understood in teleosts. In the current study, we evaluated dynamic expression profiles of lepa, lepb, and lepr mRNAs during various developmental stages in this species. Quantitative RT-PCR analysis indicated that both ligand (lepa and lepb) and receptor (lepr) genes were detected in unfertilized eggs and during embryogenesis but with different expression profiles. In addition, lepa, lepb, and lepr transcripts levels increased significantly during larval development, reaching the peak at 10, 25, and 30 days post-hatching (dph), respectively. On the other hand, changes in mRNA expression of these three genes at the brain-pituitary-gonad (BPG) axis were also investigated during ovarian maturation, and lepa, lepb, and lepr mRNAs varied greatly. Taken together, our results encompass the first study reporting the dynamic expression patterns of leptin and its receptor mRNAs in the order Pleuronectiformes, providing evidence that the leptin system could be functional and play important roles during early development and ovarian maturation in tongue sole.

Mechanisms of gene rearrangement in 13 bothids based on comparison with a newly completed mitogenome of the threespot flounder, Grammatobothus polyophthalmus (Pleuronectiformes: Bothidae)

BackgroundThe mitogenomes of 12 teleost fish of the bothid family (order Pleuronectiformes) indicated that the genomic-scale rearrangements characterized in previous work. A novel mechanism of genomic rearrangement called the Dimer-Mitogenome and Non-Random Loss (DMNL) model was used to account for the rearrangement found in one of these bothids, Crossorhombus azureus.ResultsThe 18,170 bp mitogenome of G. polyophthalmus contains 37 genes, two control regions (CRs), and the origin of replication of the L-strand (OL). This mitogenome is characterized by genomic-scale rearrangements: genes located on the L-strand are grouped in an 8-gene cluster (Q-A-C-Y-S1-ND6-E-P) that does not include tRNA-N; genes found on the H-strand are grouped together (F-12S … CytB-T) except for tRNA-D that was translocated inside the 8-gene L-strand cluster. Compared to non-rearranged mitogenomes of teleost fishes, gene organization in the mitogenome of G. polyophthalmus and in that of the other 12 bothids characterized thus far is very similar. These rearrangements could be sorted into four types (Type I, II, III and IV), differing in the particular combination of the CR, tRNA-D gene and 8-gene cluster and the shuffling of tRNA-V. The DMNL model was used to account for all but one gene rearrangement found in all 13 bothid mitogenomes. Translocation of tRNA-D most likely occurred after the DMNL process in 10 bothid mitogenomes and could have occurred either before or after DMNL in the three other species. During the DMNL process, the tRNA-N gene was retained rather than the expected tRNA-N′ gene. tRNA-N appears to assist in or act as OL function when the OL secondary structure could not be formed from intergenic sequences. A striking finding was that each of the non-transcribed genes has degenerated to a shorter intergenic spacer during the DMNL process. These findings highlight a rare phenomenon in teleost fish.ConclusionsThis result provides significant evidence to support the existence of dynamic dimeric mitogenomes and the DMNL model as the mechanism of gene rearrangement in bothid mitogenomes, which not only promotes the understanding of mitogenome structural diversity, but also sheds light on mechanisms of mitochondrial genome rearrangement and replication.

Diversity and Similarity of Flatfishes (Order- Pleuronectiformes) in Mon State, Myanmar

The samples of flatfishes were collected along the Mon State from July to December 2017 including Mawlamyine, Kyaikkhami, Setse, Zee-Phyu-Thaung Belugyun island, Paung and Thaton. The total 21 species of flatfishes were recorded in Mon State. Moreover, the relative abundance, diversity and similarity were conducted using Simpson Index, Shannon index and Jacquard’s index etc. According to the present study, the Cynoglossidae was the most dominant family and the most dominant species was Cynoglossusarelfor the present study. The relative abundance of Mawlamyine and November was relatively the highest. Furthermore, Mawlamyine was the highest diversity and Thaton was the lowest diversity. The JI value between Mawlamyine and Paung is the highest while it is the lowest between Setse and Thaton. Likewise, the JI value between August and September and the value between September and November are the same as well as the highest while it is the lowest for the comparison between July and August. The present study also expects to become a basic informational providing data for further observing (in Academic) on Order Pleuronectiformes (flatfishes) along the Mon State.

Leptin and leptin receptor genes in tongue sole (Cynoglossus semilaevis): Molecular cloning, tissue distribution and differential regulation of these genes by sex steroids

Leptin (Lep) is a key factor for the regulation of food intake and energy homeostasis in mammals. To date, a number of studies have provided evidence for the existence of multiple leptin genes in teleosts, but not much information is available in fish regarding the regulation of leptin genes by sex steriods. As a first step, two leptin genes (lepa and lepb) and a leptin receptor (lepr) gene were cloned from the half-smooth tongue sole (Cynoglossus semilaevis), a representative species of the order Pleuronectiformes. The full-length cDNAs of tongue sole lepa and lepb were 1265 bp and 1157 bp in length, encoding for proteins of 160 aa and 158 aa, respectively. The three-dimensional structures modeling of tongue sole LepA and LepB showed strong conservation of tertiary structure with other vertebrates. The full-length cDNA of tongue sole lepr was 4576 bp, encoding a protein of 1133 aa which contained all functionally important domains conserved among vertebrate LepRs. Tissue distribution analysis showed that tongue sole lepa mRNA was highly detectable in the ovary and brain, while lepb mRNA was ubiquitously expressed in various tissues. Notably, the tongue sole lepr mRNA was most abundant in the ovary. Using a primary hepatocyte culture system, we evaluated the effects of sex steroids on lep/lepr gene expression. Both 17β-estradiol (E2) and testosterone (T) inhibited hepatic lepa and lepr mRNAs without affecting lepb mRNA levels. In addition, T also suppressed growth hormone receptor 1 (ghr1), ghr2, and insulin-like growth factor 2 (igf-2) mRNA levels, and stimulated expression of igf-1 gene. On the other hand, none of these four genes were altered by E2. To the best of our knowledge, this is the first description of a direct and differential regulation of lep/lepr gene expression by sex steroids at the hepatocyte level of a flatfish, supporting that individual leptin peptide may possess different biological roles in teleosts.

Characterization of LPXRFa receptor in the half-smooth tongue sole (Cynoglossus semilaevis): Molecular cloning, expression profiles, and differential activation of signaling pathways by LPXRFa peptides

Gonadotropin-inhibitory hormone (GnIH), a novel hypothalamic neuropeptide, serves as a key player in the regulation of reproduction across vertebrates, acting on the brain and pituitary to modulate reproductive physiology and behavior. However, little information is available in teleosts regarding the intracellular signal transduction pathway in response to GnIH. To this end, we first cloned the gene of LPXRFa (the piscine ortholog of GnIH) receptor in the half-smooth tongue sole (Cynoglossus semilaevis), a representative species of the order Pleuronectiformes. The full-length cDNA of LPXRFa receptor was 2201 bp in size with an open reading frame (ORF) of 1365 bp that encoded 454 amino acids. Tissue distribution showed that LPXRFa receptor transcripts could be detected at high levels in the brain, to a lesser extent in the pituitary, and at low levels in the ovary and other peripheral tissues. In vitro functional analysis revealed that putative tongue sole LPXRFa-1 and LPXRFa-2 peptides significantly stimulated serum responsive element-dependent luciferase (SRE-luc) activity in COS-7 cells transfected with the novel receptor, and these stimulatory effects were evidently reduced by two inhibitors of the PLC/PKC pathway. In addition, neither LPXRFa-1 nor LPXRFa-2 altered the cAMP-responsive element (CRE)-luc activity, but only LPXRFa-2 could markedly decrease forskolin-induced CRE-luc activity in COS-7 cells expressing its cognate receptor. Taken together, our results encompass the first study reporting the existence of LPXRFa receptor in the order Pleuronectiformes and provide novel evidence of differential activation of signaling pathways by LPXRFa peptides in fish.

Comparative expression patterns of Sox2 and Sox19 genes in the forebrain of developing and adult turbot (Scophthalmus maximus).

The turbot, Scophthalmus maximus, belongs to the flatfishes (order Pleuronectiformes), which display substantial asymmetry of the olfactory organs and forebrain. Sox genes code for SRY-related HMG domain-bearing transcription factors involved in various developmental processes. Group B1 Sox genes as Sox2 and Sox19 appear to play major roles in neural development. Here, we characterized by in situ hybridization the developmental expression of Sox2 and Sox19 genes in metamorphic and postmetamorphic specimens and young adults of both sexes. Expression of S. maximus Sox2 (Sm-Sox2) and Sm-Sox19 mRNAs was detected in ependymal cells of different regions of the telencephalon, preoptic region, hypothalamus, and thalamus at all stages investigated. Sm-Sox2 expression but not Sm-Sox19 occurred in neurons located in particular regions such as the dorsal nucleus of the ventral telencephalon, the medial zone of the dorsal telencephalon, preoptic area and hypothalamus. Although Sm-Sox2 and Sm-Sox19 are expressed differentially in gonads, no sex differences in their expression were observed between male and female forebrains. We also investigated the topographical relation between Sox expression and cell proliferation using series double immunostained for a radial glial marker (BLBP) and cell proliferation marker (PCNA). Sm-Sox2 and Sm-Sox19 were strongly expressed in ependymal cells located in neurogenic niches revealed by the BLBP and PCNA immunostaining. Comparison with other teleosts indicates similar expression of Sox2 and Sox19 in the telencephalon, supporting conserved roles for both genes in teleost brains.

Molecular characterization and expression profiles of LPXRFa at the brain-pituitary-gonad axis of half-smooth tongue sole (Cynoglossus semilaevis) during ovarian maturation.

Gonadotropin-inhibitory hormone (GnIH) has been characterized by its ability to inhibit either basal or gonadotropin-releasing hormone (GnRH)-induced gonadotropin synthesis and release in birds and mammals. However, the physiological role of GnIH on the reproductive axis in fish remains inconclusive, with most studies focusing on the orders Cypriniformes and Perciformes. To gain insight into the role of GnIH in the regulation of reproduction in the order Pleuronectiformes, we first cloned the LPXRFa gene, the piscine ortholog of GnIH, in the half-smooth tongue sole. The full-length cDNA of LPXRFa was 918bp in size with an open reading frame (ORF) of 585bp that encoded a 194 amino acids preprohormone with a calculated molecular mass and isoelectric point of 21.73kDa and 6.52, respectively. The LPXRFa precursor encoded two putative peptide sequences that included -MPMRF or -MPQRF motifs at the C-terminal. Tissue distribution analysis showed that LPXRFa transcripts could be detected at high levels in the brains of both sexes and to a lesser extent in the ovary, heart and stomach of females, while a noteworthy expression was observed in the kidney and muscle of males. Furthermore, the expression patterns of LPXRFa mRNA during ovarian maturation were also investigated. In the brain, the mRNA expression of LPXRFa increased significantly at stage III, declined at stage V and reached a maximum at stage VI. In the pituitary, the levels of LPXRFa mRNA remained stable during ovarian maturation and increased significantly to the top level at stage V and then declined back to basal levels. In contrast, the ovarian LPXRFa mRNA levels declined sharply at stage III and remained depressed over the course of ovarian maturation. Taken together, our results provide further evidence for the existence of LPXRFa in the order Pleuronectiformes and suggest its possible involvement in the regulation of reproduction in the female tongue sole.

Ebb Light, Flood Light: Red Light, Green Light for Larval Flatfishes

Flatfishes such as flounders belong to order Pleuronectiformes and are an interesting topic for K–12 activities because of their unusual shape, and they can be used to teach students about animal movement patterns as well as ecology. Some flatfishes use the flood and ebb tidal currents to travel into and remain in small estuarine creeks because the larvae cannot swim against currents. In this activity, students play the game Red Light, Green Light to learn about larval flatfishes and their use of position in the water column, as well as tides and currents, for movement within an estuary. Students also put this knowledge to practice by drawing pictures and answering questions about larval flatfishes and their movement. This activity addresses the Next Generation Science Standard of using a model to show the relationship between animals and their habitat. Impact Statement We created an original activity to teach early elementary students about movement patterns of animals. Terms such as tides, estuaries, flounder, larvae, and metamorphosis are defined to prepare students for more complex science. Science is often not a heavy focus in earlier grade levels, but it is possible to use active outdoor games (invoking multisensory learning through seeing and doing) to teach Next Generation Science Standards such as the relationship between animals and their habitat.

Molecular characterization of Kiss2 receptor and in vitro effects of Kiss2 on reproduction-related gene expression in the hypothalamus of half-smooth tongue sole (Cynoglossus semilaevis)

Kisspeptin (Kiss) and its receptor, KissR (previously known as GPR54), play a critical role in the control of reproduction and puberty onset in mammals. Additionally, a number of studies have provided evidence of the existence of multiple Kiss/KissR systems in teleosts, but the physiological relevance and functions of these kisspeptin forms (Kiss1 and Kiss2) still remain to be investigated. To this end, we examined the direct actions of Kiss2 on hypothalamic functions in the half-smooth tongue sole (Cynoglossus semilaevis), a representative species of the order Pleuronectiformes. As a first step, the full-length cDNA for kiss2r was identified and kiss2r transcripts were shown to be widely expressed in various tissues, notably in the brain of tongue sole. Then, the effects of Kiss2 decapeptide on reproduction-related gene expression were evaluated using a primary hypothalamus culture system. Our results showed that neither gnrh2 nor gnrh3 mRNA levels were altered by Kiss2. However, Kiss2 significantly increased the amounts of gnih and kiss2 mRNAs. In contrast, Kiss2 elicited an evident inhibitory effect on both gnihr and kiss2r mRNA levels. To the best of our knowledge, this is the first description of a direct and differential regulation of reproduction-related gene expression by Kiss2 at the hypothalamus level of a teleost fish. Overall, this study provides novel information on the role of Kiss2/Kiss2R system in the reproductive function of teleosts.

Environmental drivers of the distribution of the order Pleuronectiformes in the Northern Spanish Shelf

A total of 17 species of the order Pleuronectiformes were caught in standardized groundfish surveys in the Cantabrian Sea and Galician waters between 2002 and 2011. The abundance and distribution of the dominant flatfish species were analysed, as well as drivers that affected the distribution. The results of a hierarchical classification analysis indicated three primary assemblages: (1) an inner shelf group (~50–100m), characterized by Pegusa lascaris, Buglossidium luteum and Solea solea; (2) a medium and outer shelf group (~125–250m), represented by Arnoglossus laterna, A. imperialis, Microchirus variegatus, Lepidorhombus whiffiagonis and L. boscii; and (3) a shelf break and upper slope group (~325–450m), formed by Bathysolea profundicola. MANOVA analyses indicated that depth was the primary structuring factor, although sediment size and geographical position were also important. A bathymetric segregation pattern was drawn and indicated species replacement with increasing depth irrespective of size. Most of the species appeared to be associated with fine sand, except A. imperialis, which showed more association with coarse sand. The most abundant species were L. boscii, M. variegatus and A. laterna, which also had the widest and most ubiquitous distribution. They had a clear distribution farther west, whereas the rest seemed to be a more eastern group, and L. whiffiagonis, B. profundicola and A. imperialis were present at more northern latitudes. A. laterna was the most abundant species in the Galician shelf. In general, flatfish species in the northern Spanish shelf go through a slight ontogenetic niche shift primarily as a function of depth and sediment size. Smaller individuals tend to occur in shallower waters than their larger counterparts, except for A. laterna, M. variegatus and L. boscii, which have the opposite behaviour. Most flatfish show an affinity for a larger sand grain size with growth.

Centromeric Satellite DNA in Flatfish (Order Pleuronectiformes) and Its Relation to Speciation Processes.

Two new centromeric satellite DNAs in flatfish (Order Pleuronectiformes) have been characterized. The SacI-family from Hippoglossus hippoglossus, restricted to this species, had a monomeric size of 334 base pair (bp) and was located in most of the centromeres of its karyotype. The PvuII-family, with a monomeric size of 177 bp, was initially isolated from the genome of Solea senegalensis, and fluorescent in situ hybridization (FISH) localized the repeat to centromeres of most of the chromosomes. This family could only be amplified in 2 other species of the genus Solea (Solea solea and Solea lascaris). Molecular features and chromosomal location indicated a possible structural and/or functional role of these sequence repeats. The presence of species-specific satellite-DNA families in the centromeres and their possible role in the speciation processes in this group of fishes is discussed.

Brandt's cormorant diet (1994–2012) indicates the importance of fall ocean conditions for northern anchovy in central California

AbstractEffective ecosystem‐based management requires a comprehensive understanding of the functional links in the system. In many marine systems, forage species constitute a critical link between primary production and upper trophic level marine predators. As top predators, seabirds can be indicators of the forage species they consume and the ocean processes that influence these populations. We analyzed the diet and breeding success for the years 1994, 2003, 2005, and 2007–2012 of the Brandt's cormorant (Phalacrocorax penicillatus), a piscivorous diving seabird, breeding in central California, to evaluate the extent to which cormorant diet composition relates to prey availability, and how diet composition relates to breeding success and ocean conditions. Cormorant diet was primarily composed of young‐of‐the‐year (YOY) northern anchovy (Engraulis mordax), YOY rockfish (Sebastes spp.), and several species of small flatfish (order Pleuronectiformes). YOY rockfish consumption was positively related to their abundance as measured in a late spring pelagic midwater trawl survey. Northern anchovy appeared to be the most important prey as its consumption was positively related to cormorant breeding success. More northern anchovy were consumed in years where warm‐water conditions prevailed in the fall season before cormorant breeding. Thus, warm ocean conditions in the fall appear to be an important contributing factor in producing a strong year‐class of northern anchovy in central California and consequently a strong‐year class of Brandt's cormorant on the Farallon Islands.

The complete mitochondrial genome of Pseudaesopia japonica (Pleuronectiformes: Soleidae)

The Pseudaesopia japonica belongs to family Soleidae in order Pleuronectiformes. In this study, the complete mitochondrial genome of P. japonica was determined and described. The mitogenome is 16 790 bp in length and consists of 13 protein-coding genes, 22 tRNAs, two rRNAs, one control region, and a light strand replication origin. The arrangement of this mitogenome is identical to that of the typical teleost. The overall base composition is 28.5%, 25.7%, 30.3%, and 15.5%, for A, T, C, and G, respectively, with a slight bias on A + T content (54.3%). The maximum likelihood phylogeny tree of 22 flatfishes demonstrated that the species from Zebrias and Aesopia firstly formed a sister group, and then clustered in the same clade with P. japonicas. This study is expected to contributing to the systematic evolution of P. japonicas and further phylogenetic relationship of Soleidae and Pleuronectiformes.

Acanthocephalan Cystacanths from Flatfish (Order Pleuronectiformes) in Tropical Australian Waters.

Cystacanths of 4 species of Acanthocephala are reported for the first time from various species of fish belonging to the Order Pleuronectiformes from waters of the western Gulf of Carpentaria and the central coast of Queensland, Australia: Corynosoma cetaceum Johnston and Best, 1942 (Family Polymorphidae), Serrasentis cf. sagittifer (Linton, 1889) and Rhadinorhynchus sp. (Family Rhadinorhynchidae), and Gorgorhynchoides sp. (Family Isthmosacanthidae). Approximately 32% of the 515 individual fish belonging to 24 species were infected with at least 1 cystacanth. Serrasentis cf. sagittifer was the most-commonly encountered, infecting a total of 18 species of fish across both regions. Gorgorhynchoides sp. infected 7 fish species in the Gulf of Carpentaria only while Rhadinorhynchus sp. (1 fish species) and C. cetaceum (7 fish species) were only found on the central coast of Queensland. Most fish were infected with a single cystacanth of any species. There was no relationship between total length of fish and intensity of infection for any species. This paper provides information on parasite infections in fish hosts commonly caught as by-catch and outlines the need for further studies on these fish to be able to determine sustainability of such fish stocks.

Comparison of Hatching Mode in Pelagic and Demersal Eggs of Two Closely Related Species in the Order Pleuronectiformes

We compared several characteristics of the pelagic eggs of Verasper variegatus with those of demersal eggs of Pseudopleuronectes yokohamae, both in the order Pleuronectiformes (halibuts or flatfishes). V. variegatus eggs had about twice the diameter of P. yokohamae eggs. However, the total egg protein weight of P. yokohamae was similar to that of V. variegatus. The specific gravity of P. yokohamae eggs was calculated to be 7-fold that of V. variegatus. The difference in size is the main feature distinguishing the two types of egg. The thickness of the egg envelope of P. yokohamae- more than twice that of V. variegatus-must affect the manner of hatching. The amount of hatching enzyme synthesized in pre-hatching embryo was estimated to be larger in P. yokohamae than V. variegatus. The distribution of hatching gland cells differed between the species. In V. variegates embryos, these were located on the yolk sac as a narrow ring-shaped belt, resulting in cleavage of the egg envelope into two parts by digesting a limited region of the egg envelope, called "rim-hatching". The hatching gland cells of P. yokohamae embryos were distributed all over the surface of the yolk sac, forming a hole through which the embryo could escape. Thus, the location of the hatching gland cells in pre-hatching embryos varied during the evolution of the Pleuronectiformes, depending on the egg type and manner of hatching.

Changes in forage fish community indicated by the diet of the Brandt's cormorant (Phalacrocorax penicillatus) in the central California Current

The population, productivity and diet of two Brandt's cormorant (Phalacrocorax penicillatus) colonies located in the central California Current were compared. The offshore colony on Southeast Farallon Island has experienced a declining population over time and anomalously low productivity in recent years. The nearshore colony near Point Arguello has been increasing and its productivity has remained stable. The diets of cormorants at the two colonies elucidated by analysis of regurgitated pellets, while different, have shown similar decreases in the consumption of northern anchovy (Engraulis mordax) since 2008, followed by increased consumption of rockfish (Sebastes spp.) and flatfish (order Pleuronectiformes). By using the diet results from another seabird nesting in central California, the rhinoceros auklet (Cerorhinca monocerata), and one from which whole fish can be obtained, we found that the rockfish species assemblage has changed with offshore rockfish species decreasing while nearshore ones have increased. This change in the rockfish species has negatively impacted Brandt's cormorants at the offshore colony by forcing them to make longer foraging trips to meet energy needs of themselves and their chicks; this has led to low breeding success and a declining population at this site. On the other hand, the nearshore colony has abundant nearby food resources, and it has prospered. These results underscore the value of using seabird data from multiple colonies to better understand changes occurring in the marine environment.

Phylogenetic relationships of Russian far eastern flatfish (Pleuronectiformes, Pleuronectidae) based on two mitochondrial gene sequences, Co-1 and Cyt-b, with inferences in order phylogeny using complete mitogenome data

The systematics and phylogeny of flatfish is investigated on the complete sequence of nucleotides at subunit 1 cytochrome c oxidase (Co-1) and cytochrome b (Cyt-b) genes. In total 17 species from our collection and some species from GenBank were analyzed. Four types of trees were built: Bayesian (BA), maximum likelihood (ML), maximum parsimony (MP), and neighbor joining (NJ). These trees showed similar topology. Two separate clusters on the trees support subfamily Hippoglossoidinae and Hippoglossinae subdivision and monophyletic status of these taxa. The subfamily Pleuronectinae also can be considered monophyletic, if the tribe Microstomini is excluded from it and genus Lepidopsetta is moved into the tribe Pleuronectini. Mitogenomes represented by 25 complete sequences from NCBI GenBank were analyzed. After alignment two sets of nucleotide sequences were formed and investigated independently. One set included 13 structural genes (14,886 bp), the second set comprised by the mtDNA without ND6 gene (10,457 bp). Both data sets give congruent phylogenetic signal that agreed with conventional views on the taxonomy of the order Pleuronectiformes; however, the first set gives better topology. In BA gene tree there are two well supported nodes which include the representatives of suborders Pleuronectoidei and Psettoidei. Within Pleuronectoidei two superfamilies, Pleuronectoidea and Soleidea are highly supported in BA but in all four kinds of gene trees (BA, ML, MP and NJ) the only superfamily Pleuronectoidea is well supported. At the top of hierarchy, all flatfishes belonging to the order Pleuronectiformes forming also a monophyletic clade in our data, with support level of 100% but in BA tree only. The monophyly of the family Pleuronectidae is well supported both by single gene data and by complete mtDNA sequences.