Allozyme and mitochondrial DNA variation in orange roughy,Hoplostethus atlanticus (Teleostei: Trachichthyidae): little differentiation between Australian and North Atlantic populations

Allozyme and mitochondrial DNA (mtDNA) genetic variation was compared in orange roughy (Hoplostethus atlanticus Collett) collected from waters off southern Australia and from waters about 22 000 km away in the North Atlantic west of Scotland. Samples were screened for 11 polymorphic allozyme loci and with 9 restriction enzymes. Significant heterogeneity between the two areas was detected for three allozyme loci (ADA ^*, CK ^* and GPI-1 ^*), and the overall G _ST (gene-diversity statistic) value of 1% was small but significant. Significant mtDNA haplotype heterogeneity was observed after ²- of haplotype frequencies but not after a G _ST analysis. Nucleotide sequence-diversity analysis showed very low net divergence (0.0023%) between the two samples. The Australian orange roughy had a lower allozyme heterozygosity and a lower mitochondrial DNA nucleon diversity than the North Atlantic sample. The very limited, although significant, allozyme and mitochondrial DNA heterogeneity between these areas suggests that there is some gene flow between these two populations. The species appears to be widespread, with its presence reported from the southern Pacific, southern Indian, and northern and southern Atlantic Oceans, and it is likely that gene flow between the antipodes is mediated by stepping-stone exchange between adjacent populations rather than by direct migration.     

Diet of orange roughy,Hoplostethus atlanticus (Pisces: Trachichthyidae) on the Challenger Plateau,New Zealand

Stomach contents of 687 orange roughy [Hoplostethus atlanticus (Collett, 1889)] from the Challenger Plateau, sampled in March, July and November 1984, were examined. The relative importance of different components of the diet was assessed using three feeding indices which combine, in different ways, percentage frequency of occurrence, percentage number, and percentage weight of prey categories. For both sexes, for all size classes and at any time of the year, the most frequent and abundant components of the diet were prawns, followed by fish, squid, amphipods and mysids. The main natant decapod families were Oplophoridae, Pasiphaeidae, and Sergestidae. Fish, mainly Chauliodontidae and Myctophidae, were the most important prey by weight. Most prey species were benthopelagic and mesopelagic organisms which move towards the surface at night. However, H. atlanticus can be caught by bottom trawl (between 750 and 1 200 m) during any 24 h period, and there was no evidence of vertical migration in search of their prey. When the fish grow in length, there is a transition in the diet from prawns, mysids, and fish, to prawns, fish and squid. Squid were not found in the stomach contents of fish smaller than 20 cm. These dietary changes may be linked to modifications in morphology with growth. The relative proportions of the main dietary items, and in particular the natant decapod families, varied with time of the year. Also, the stomach data seem to indicate an increasing importance of fish and squid in the diet in deeper water. H. atlanticus appears to be an opportunistic predator, consuming a wide variety of invertebrates and fishes. Our results provide evidence to support the notion that benthopelagic predators which consume vertically migrating mesopelagic fish, have an important role in the transfer of energy to the benthos.     

Age determination of orange roughy,Hoplostethus atlanticus (Pisces: Trachichthyidae) using210Pb:226Ra disequilibria

Natural levels of²¹⁰Pb:²²⁶Ra in otoliths of orange roughy,Hoplostethus atlanticus, from south-east Australian waters, were measured to determine fish ages radiometrically. Up to maturity, radiometric age estimates were consistent with a single constant otolith growth rate. Radiometric ages for juveniles were comparable with, but greater than, those obtained in a recent, validated New Zealand study which employed counts of annuli on the surface of otoliths. Beyond maturity, radiometric ages were obtained by modelling with an otolith growth rate set at 45% of the juvenile rate. Radiometric ageing confirms that orange roughy is very slow-growing, with an age at maturity (32 cm standard length, SL) of ~ 32 yr, and is very long-lived, with fish 38 to 40 cm being 77 to 149 yr old. These results have important implications for the management of the fishery.     

Stock discrimination of orange roughy,Hoplostethus atlanticus,by parasite analysis

Seasonal changes of respiratory metabolism and biochemical composition were studied in Actinia equina L. (Cnidaria: Anthozoa) collected in 1982 and 1983 from two shore locations in Spain. Changes of metabolic activity follow closely those of biochemical constituents, particularly carbohydrates and lipids, showing maxima in late fall and spring. Weight dependence, established according to the exponential equation y=a x^b (x=weight), results in virtually isometric relationships for proteins, total lipids and sugars but not glycogen. Allometry for this last compound represents a reduction in glycogen level in connection with size decrease; this weight effect declines when glycogen levels rise, both in field and laboratory fed anemones, thus suggesting a stronger influence of food availability on storage in small anemones.     

Morphometric and meristic comparison of orange roughy (Hoplostethus atlanticus: Trachichthyidae) from the Puysegur Bank and Lord Howe Rise,New Zealand,and its implications for stock structure

Significant morphological differences were found between orange roughy, Hoplostethus atlanticus, caught in 1993 on the Puysegur Bank and those caught on the Lord Howe Rise, New Zealand. A total of 17 separate body measurements and counts were taken from each fish. Treating the sexes separately, significant differences were found between sites when linear relations between eight particular body measures and standard length were compared. The eight measures indicating differences in shape were head length, snout length, orbit diameter, maxilla width, premaxilla length, caudal peduncle, gill raker count, and anal fin count. Discriminant functions were used to compare sites and sexes, and these permitted a >90% success rate in distinguishing fish from the two sites. A discriminant function could not be found which successfully separated two separate tows made on the Puysegur Bank, indicating that phenotypic differences do not vary as much within a site as they do between sites. All analyses made suggested that fish from the two different areas had different body shapes. Morphological differences, such as those found in the current investigation, could only arise if the fish populations from the two areas were relatively discrete. There may be some movement of fish between areas, quite possibly enough to bring about relative genetic homogeneity, but clearly there is insufficient to prevent the two populations from differing phenotypically. These differences suggest that morphometric analysis may be a useful tool for distinguishing New Zealand orange roughy stocks.     

DNA fingerprints of orange roughy,Hoplostethus atlanticus: a population comparison

Genetic variability among Hoplostethus atlanticus collected from two spawning grounds east and west of New Zealand was examined using DNA fingerprints as revealed by hybridization with three clonal probes: 33.15, M13 and 3HVR. The 33.15 and 3HVR fingerprints revealed a complex pattern of restriction fragments, apparently refecting a multi-locus system of highly variable minisatellite alleles similar to the pattern of alleles reported in other vertebrates. The M13 fingerprints revealed a distinct pattern of restriction fragments of high molecular weight, reflecting a single-locus system that overlapped with the family of minisatellite alleles observed in 33.15 fingerprints. In a sample of 12 orange roughy collected on a single regional spawning site, the average percent similarity of 33.15 fingerprints was 21.15% (SD=17.75), the average percent similarity of 3HVR fingerprints was 14.32% (SD=14.45) and the inferred average allelic frequency of the M13 single-locus system was 0.071. A comparison of 33.15 and M13 fingerprints from two distant spawning sites ground New Zealand revealed no obvious regional differences. The variability of orange roughy fingerprints was so great, however, that regional comparisons could not be considered conclusive indicators of genetic identity. Our results provide a preliminary assessment of the power and pitfalls of using DNA-level markers for the population analysis of marine fish.     

Population variation in the mitochondrial cytochrome b gene of the orange roughy Hoplostethus atlanticus and the hoki Macruronus novaezelandiae

To describe the genetic relationship among regional populations of two commercially valuable species of marine fish, the orange roughy Hoplostethus atlanticus and the hoki Macruronus novaezelandiae, the mitochondrial (mt) DNA haplotypes of 59 individuals were defined by direct sequencing of the cytochrome b gene. Samples of orange roughy were collected on four fishing grounds around New Zealand, one off the west coast of Tasmania and one near South Africa from 1990–1991. Samples from hoki were collected on three fishing grounds around New Zealand and one off Tasmania during the same period. An average of 252 nucleotides were sequenced from each of 32 orange roughy and an average of 372 nucleotides from each of 27 hoki. Sequence variation allowed the definition of 9 unique orange roughy haplotypes and 5 hoki haplotypes. Genetic variation, as measured by both average sequence divergence and haplotype diversity, was high in the orange roughy (nucleotide diversity=0.590%, haplotype diversity=0.782) and low in the hoki (nucleotide diversity=0.076%, haplotype diversity=0.279) relative to a similar survey of the Atlantic cod, Gadus morhua. Differences in haplotype frequencies of orange roughy from New Zealand, Tasmania and South Africa were not significant, and the most common haplotype was found in similar frequencies in these three geographically distant regions. Differences in haplotype frequencies between the New Zealand and Tasmanian samples of hoki were significant, suggesting restricted gene flow between these two regions. The contrasting patterns of low but regionally subdivided genetic variation in the hoki versus high but geographically undifferentiated genetic variation in the orange roughy may be attributed to the low fecundity, slow maturation and long lifespan of the orange roughy relative to the hoki.     

Population structure and life history of orange roughy (Hoplostethus atlanticus) in the SW Pacific: inferences from otolith chemistry

We examined site differences in the elemental composition of the primordium and ontogenetic variability of Sr in otoliths of fish from Australia and New Zealand and, as an out-group, the North Atlantic. Differences among sites in primordium composition are slight, but significant for all five elements assayed (Sr, Pb, Cu, Zn and Hg), but principally reflect differences between the North Atlantic and SW Pacific specimens, do not replicate for independent samples in the SW Pacific and constitute a poor “natural tag” in roughy, with <25% of fish successfully assigned to source location. However, mean Sr weight-fractions at the primordium showed similar latitudinal variation across sites in Australia, New Zealand and the Tasman Sea, indicating both spatially structured populations and a common structuring process across the region. Comparisons of ontogenetic variability of Sr in otoliths from juveniles and young adults within and between sites in the SW Pacific strongly support the hypothesis that variability in this element is site-specific and environmentally sensitive, although the environmental factors involved are not obvious. The otolith analysis confirms previous suggestions that juvenile and adult Hoplostethus atlanticus are relatively sedentary, but also indicates that the population sub-structuring by age within sites is more complex and there are likely to be more spawning areas in Australian waters than previously thought. More broadly, although single point analysis of otolith composition at the primordium resolves a population structure in roughy, alone it is not precise enough to test hypotheses about the processes causing this structure. Ontogenetic variability in Sr provides better resolution of spatial structure, even in a relatively homogenous marine environment like the deep ocean, and also provides insight into behavioural and ecological factors. Ontogenetic analyses of Sr in otoliths are expensive to obtain, require more effort in specimen preparation than single point analyses, and are difficult to compare statistically, but the increased information they yield warrants their broader consideration in marine species.     

Transition zone in otoliths of orange roughy (Hoplostethus atlanticus) and its relationship to the onset of maturity

A transition zone in the otoliths of orange roughy (Hoplostethus atlanticus) is described where annulus width decreases markedly. For fish sampled at spawning time, the relationship between gonad stage and the presence or absence of a transition zone is consistent with the hypothesis that this zone marks the onset of maturity, if allowance is made for some error in gonad staging and identification of the zone. Another data set from a non-spawning area after the spawning season is not consistent with the hypothesis. However, these latter data are considered less reliable because of the small size of the fish in the sample and difficulties in determining, outside the spawning season, whether a fish has been reproductively active. This hypothesis was used to provide estimates of the median age (30 yr) and length (31 cm) at the onset of maturity for the Chatham Rise, New Zealand population. These estimates are unaffected by the biases associated with the usual ogive method of estimation. Received: 27 June 1997 / Accepted: 1 July 1997     

Between-area differences in age and length at first maturity of the orange roughy Hoplostethus atlanticus

Mean age and length at onset of maturity were estimated for orange roughy (Hoplostethus atlanticus) populations off New Zealand, Namibia, and Great Britain. Significant between-area differences were apparent in both these parameters. Implications of the between-area differences on stock structure of orange roughy in New Zealand waters are discussed. A proportional relationship between age at onset of maturity and modal size of fish in the mature population is demonstrated. This is consistent with later-maturing fish experiencing a longer period at a pre-maturity growth rate that is relatively faster than the rate of somatic growth after maturity. Received: 22 December 1997 / Accepted: 5 May 1998     

Role of wax esters in determining buoyancy inHoplostethus atlanticus (Beryciformes: Trachichthyidae)

Specimens of the orange roughyHoplostethus atlanticus were caught at the edge of the Chatham Rise, east of the South Island of New Zealand, in June 1988, during a commercial fishing voyage. Several tissues of the fish were found to contain large quantities of lipid almost entirely as wax esters. These tissues include a fat-invested swimbladder, the skin, skull and spine, and a fat-infiltrated tissue found in the neurocranial cavity. The physical properties of the wax esters have been examined relative to the temperatures that exist where the fish are found. Differential scanning calorimetry and NMR spectroscopy both suggest that at 6°C, the temperature at which the fish are located in the water column, the lipid is partially solid. However, at 14°C, the temperature at the surface where the fish are positively buoyant, the lipid is completely liquid. It is proposed that the increased density of the lipid at 6°C would make the fish neutrally buoyant in its normal habitat. The location of the lipidrich tissues has been examined relative to their possible buoyancy contribution, and a histological examination of the swimbladder has been performed.     

Genetically isolated stocks of orange roughy (Hoplostethus atlanticus), but not of hoki (Macruronus novaezelandiae), in the Tasman Sea and Southwest Pacific Ocean around New Zealand

Restriction fragment-length polymorphisms of mitochondrial (mt) DNA, generated by six-base restriction enzymes, were used to examine the stock structure of the two major commercial species in New Zealand fisheries, orange roughy (Hoplostethus atlanticus Collett) and hoki (Macruronus novaezelandiae Hector). Samples of orange roughy from six spawning sites around New Zealand and one site off Tasmania show significant genetic heterogeneity. Samples from two sites around the south of New Zealand were genetically similar to each other, but dissimilar to samples from all other sites. The common haplotype observed in northern sites was absent from the two southern sites. Low numbers of unique haplotypes were restricted to the Challenger Plateau in the Tasman Sea. There was no significant genetic heterogeneity in five samples of hoki from two spawning sites off New Zealand and one site off Tasmania.     

Genetic stock structure of the swordfish (Xiphias gladius ) inferred by PCR-RFLP analysis of the mitochondrial DNA control region

Restriction fragment length polymorphism (RFLP) analysis was performed on PCR amplified DNA fragments containing the control region of the swordfish (Xiphias gladius Linnaeus, 1758) mitochondrial DNA. A total of 456 individuals comprising 13 local samples (six Pacific, three Atlantic, the Mediterranean Sea, two Indian Ocean and the Cape of Good Hope) were surveyed with four endonucleases (Alu I, Dde I, Hha I and Rsa I), yielding a total of 52 composite genotypes. Within-sample genotypic diversity (H) was high ranging from 0.702 to 0.962 with a value of 0.922 for the pooled sample. Significant geographic variation in the frequencies of genotypes and restriction patterns was revealed. The Mediterranean sample was highly distinct from all other samples. Further, Rsa I digestion revealed high levels of polymorphism in all but the Mediterranean samples, indicating that exogenous swordfishes rarely enter that body of water. Heterogeneity between the North and South Atlantic samples was significant, both of which differed from those of the Pacific. In contrast, the Indian Ocean samples were not significantly different from the samples of South Atlantic and Pacific. Genetic differentiation among the Pacific samples was low. The results indicate that the world-wide swordfish population is genetically structured not only among, but also within ocean basins and suggest that gene flow is restricted despite the absence of geographic barriers. Received: 28 August 1996 / Accepted: 2 October 1996     

Phylogenetic relationships of coral-associated gobies (Teleostei,Gobiidae) from the Red Sea based on mitochondrial DNA data

Bryaninops, Gobiodon, Paragobiodon and Pleurosicya are the most abundant genera of coral-associated gobies. These genera are adapted to live among coral, while other small reef gobies (e.g., the genus Eviota) show no obligate association with this living substrate. Thirteen coral-associated species and two Eviota species were sampled from different regions of the Red Sea, along with four populations/species of Gobiodon from the Indian and western Pacific Oceans. A molecular phylogenetic analysis was performed using partial sequences of 12S rRNA, 16S rRNA and cytochrome b mitochondrial genes, 1,199 base pairs in total. Several clades were consistently resolved in neighbor joining-, maximum parsimony-, maximum likelihood and Bayesian analyses. While each of the four genera Gobiodon, Paragobiodon, Bryaninops and Pleurosicya proved to be monophyletic, their relative position in the phylogeny did not support an emergence of coral-associated gobiids as a monophyletic assemblage. Instead, two separate monophyletic sub-groups were discovered, the first comprising Gobiodon and Paragobiodon, and the second Bryaninops and Pleurosicya. Our molecular phylogenetic examinations also revealed one unassigned species of Gobiodon from the Maldives as a distinct species and confirmed three putative and yet unassigned species from the Red Sea. Moreover, the uniformly black colored species of Gobiodon are not monophyletic but have evolved independently within two distinct species groups. Genetic distances were large in particular within Pleurosicya and Eviota. Estimated divergence times suggest that coral-associated gobies have diversified in parallel to their preferred host corals. In particular, divergence times of Gobiodon species closely match those estimated for their typical host coral genus Acropora.     

Allozyme and mitochondrial DNA variation in yellowfin tuna (Thunnus albacares) from the Pacific Ocean

Samples of yellowfin tuna (Thunnus albacares) collected in 1991 and 1992 from the western, central and eastern regions of the Pacific Ocean were examined for genetic variability. Four polymorphic allozyme loci (ADA ^*, FH ^*, GPI-S ^* and GPI-F ^*) were examined in all samples and a fifth polymorphism (GDA ^*) was examined in western and central samples only. Samples were also screened for mitochondrial DNA variation following restriction analysis by two enzymes (BcII and EcoRI) detecting polymorphic cut sites. Eighteen mtDNA haplotypes were revealed, with an overall nucleon diversity of 0.678. A subset of individuals screened for eight restriction enzymes had an overall nucleon diversity of 0.724 and a mean nucleotide diversity per sample of 0.359%. No significant spatial heterogeneity was detected for alleles at the ADA ^*, FH ^*, GPI-S ^* and GDA ^* loci nor for the mtDNA haplotypes. Significant heterogeneity was detected for GPI-F ^*. At this locus, the two eastern samples (southern California and northern Mexico) were not significantly different from each other but were significantly different (P<0.001) from the five western/central samples (Philippines, Coral Sea, Kiribati, Hawaii-91 and Hawaii-92). GPI-F ^* 100 was the most common allele in western and central regions, GPI-F ^* 75 the most common in eastern samples.     

Fluorescence studies of dye displacement from DNA by chromium(III) complexes: Evidence for cation induced DNA condensations

The interactions of 10 different chromium(III) complexes with isolated calf thymus DNA have been analysed by studying the electronic and fluoresence spectra of intercalated ethidiumbromide. Triply charged cationic complexes including: [Cr(urea)₆]Cl₃.3H₂O, [Cr(1,10‐phenanthroline)₃](ClO₄)₃.2H₂O, [Cr(2,2'‐bipyridyl)₃] (ClO₄)₃.2H₂O, [Cr(ethylendiamine)₃]Cl₃.3.5H₂O and [Cr(NH₃)₆](NO₃)₃ displaced the dye from DNA. Similar effects were observed in experiments using the non‐intercalating dye bisbenzimidazole ("Hoechst 33258"). However, singly charged cationic, anionic and uncharged chromium(III) complexes such as: cis‐[Cr(1,10‐phenanthroline)₂Cl₂]Cl.2H₂O, cis‐[Cr(2,2'‐bipyridyl)₂Cl₂]Cl.2H₂O, [Cr(glutathione)₂]Na₂, [Cr(cysteine)₂]Na.2H₂O and [Cr(glycine)₃] were unable to displace both ethidiumbromide and bisbenzimidazole from DNA. There was no evidence for the formation of co‐ordinate bonds between chromium(III) and DNA for any of the above complexes. The charge and type of ligand are important in controlling the interaction of chromium(III) with isolated DNA in vitro. Our findings indicate that the outer sphere interaction of a chromium(III) complex with DNA is weak and unlikely to be the mechanism by which chromate causes DNA impairments in vivo and in vitro.     

居民垃圾分类回收行为的现状描述与影响因子分析——基于广东省的实证研究

随着快速的城市化,居民生活垃圾对环境生态产生的危害越来越大,已成为影响居民生活居住环境的重要问题.探讨如何促进居民进行有效的垃圾分类回收行为显得至关重要.采用因子分析相关理论和方法,对广东省居民调查的824份数据进行了居民生活垃圾分类意识和行为现状描述与低效性分析.结果表明,影响广东省居民垃圾分类回收的6个相关的变量中...     

Close genetic similarity of Atlantic and Pacific skipjack tuna (Katsuwonus pelamis) demonstrated with restriction endonuclease analysis of mitochondrial DNA

Restriction endonuclease analysis of mitochondrial DNA indicated a surprisingly high degree of genetic similarity between skipjack tuna (Katsuwonus pelamis) from the Atlantic and Pacific Oceans. The present results (1983) support the findings of previous morphological and electrophoretic studies. Evidently, since the uplift of the Panamá land bridge about 3.1 million years ago, there has been continued genetic contact between Atlantic and Pacific skipjack tuna, presumably via the Southern Ocean.     

Demographic composition of the feeding population of juvenile loggerhead sea turtles (Caretta caretta) off Charleston,South Carolina: evidence from mitochondrial DNA markers

To determine the origin of juvenile loggerhead turtles (Caretta caretta) that occupy the Charleston Harbor Entrance Channel at Charleston, South Carolina, USA, mitochondrial DNA restriction-fragment length polymorphisms from this feeding population were compared to haplotypes from candidate nesting populations. Previous studies have defined two major nesting populations in the southeastern USA, one corresponding to Florida and the other to Georgia/South Carolina. These nesting populations are distinguished by both unique haplotypes and frequency distributions of common haplotypes. The frequency distribution of haplotypes in the juvenile feeding-ground population was significantly different from both nesting populations, implying that the feeding aggregate is drawn from two or more nesting populations. Assuming that these turtles are derived exclusively from rookeries in the southeastern USA, a maximum likelihood estimator indicates that approximately half are from the Florida rookery and half are from the northern (Georgia/South Carolina) rookery complex. Because 91% of nesting in the southeastern USA occurs in Florida rookeries and 8% in the northern complex, the 50:50 ratio indicates that juvenile turtles from Georgia and South Carolina tend to feed preferentially near their respective rookery locations. Human encroachment on this feeding habitat may pose an especially high risk to the smaller Georgia/South Carolina rookeries.