Recent advances in artificial breeding and larval rearing of silver pomfret Pampus argenteus (Euphrasen 1788) in Kuwait

During last several years, Kuwait Institute for Scientific Research (KISR), Kuwait has been trying to develop a sustainable culture technique for silver pomfret (Pampus argenteus). This paper reports the recent research advances in the breeding and rearing of silver pomfret fry at KISR. The eggs collected from wild silver pomfret during spawning seasons of 2012 to 2015 were artificially fertilized under laboratory condition. The average hatching rates of artificially fertilized eggs collected from wild silver pomfrets were 25.6%, 44.8%, 76.7%, and 53.5.0% and average survival rates of metamorphosed fry produced from these eggs were 3.7%, 5.7%, 4.4% and 3.8% for the spawning seasons 2012, 2013, 2014, and 2015, respectively. For captive brood stocks, observation on the spawning time at hourly interval by collecting eggs from the nets set at out-flow of brood tanks showed that the spawning time for captive silver pomfret starts at the time of sunset. In 2012, two groups of captive silver pomfret broods spawned a total of 62x10³ and 66x10³ eggs, but the eggs were unfertilized. No captive spawning occurred during 2013 and 2014 spawning season. In 2015, captive silver pomfret broods in three tanks spawned about 653x10³, 673x10³ and 270x10³ eggs, and in 2016, the broods in four tanks spawned about 669x10³, 22x10³, 3x10³ and 366x10³ eggs. However, from these eggs only 1,400 and 1,000 fertilized eggs were collected which produced 300 and 123 hatched larvae in July 2015 and June 2016, respectively. The larvae produced from the 2015 broods did not survive beyond 56 days of metamorphosed stage while larvae produced from 2016 captive broods survived for 9 days only. The mortality of the larvae from captive brood could be related to the poor egg or milt quality. However, efforts are continued to improve the eggs and sperm quality through proper brood management.     

Effects of low salinity on antioxidant enzymes activities in kidney and muscle of juvenile silver pomfret Pampus argenteus

Silver pomfret (Pampus argenteus) is an important fish species in China, whose fatality is strongly affected by marine salinity. To better understand the tolerance of sliver pomfret to varying environmental salinity at its early developmental stages, the influence of salinity on the activities of five antioxidant enzymes – superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX), glutathione-S-transferase (GST), and glutathione reductase (GR) – in the kidney and muscle of juvenile silver pomfret was investigated in this study. Divided groups A, B, C, D at random, the juvenile silver pomfrets were then treated with different salinity levels of 25, 20, 15, and 10 with time periods of 24, 48, 96, and 120 h respectively. Generally speaking, the results show that the renal and muscular SOD as well as the renal CAT activities first rise then drop (P < 0.05), while the muscular CAT activity increases with the elapse of treatment time (P < 0.05) and peaked at 96 h, then decreased at 120 h, while the muscular CAT activity in group D increased consistently and peaked at 120 h. The renal GPX activity decreases significantly (P < 0.05) as the salinity drops and treatment time prolongs with its lowest level occurring at 96 h in group D, while the muscular GPX activity increases first then decrease (P < 0.05); moreover the renal and muscular GPX activities vary reversely in compensation. On a whole, the renal and muscular GST and GR activities first rise then drop (P < 0.05) with the drop of salinity and elapse of time. To sum up, the results acquired from study indicate that the salinity decrease could effectively stimulate and enhance the antioxidant enzyme activity in the kidney and muscle of the juvenile silver pomfrets, thus effectively eliminating the excessive reactive oxygen species (ROS) and minimizing the body damage. Characterized by certain sequentiality and tissue specificity, the activation of the antioxidant enzymes could also be inhibited when the salinity varies beyond the tolerance range of the body.     

Isolation and characterization of microsatellites in Sparganium emersum and cross‐species amplification in the related species S. erectum

We developed seven novel polymorphic microsatellite loci for the aquatic macrophyte Sparganium emersum (Sparganiaceae). These were characterized on 62 individuals collected from nine different populations. In this set of individuals, seven to 20 alleles per locus were detected and observed heterozygosity ranged between 0.16 and 0.95. Cross‐species amplification was tested in the related species Sparganium erectum, and was successful for five of the seven microsatellite loci.     

Isolation of novel microsatellite loci in the black goby Gobius niger and cross‐amplification in other gobiid species (Perciformes,Gobiidae)

Twelve microsatellite loci were isolated from and characterized for the black goby Gobius niger. These loci were tested on a total of 48 individuals from two geographically distant locations (Orbetello and Cattolica) and the number of alleles ranged from two to 18, with expected (H_e) and observed (H_o) heterozygosities ranging from 0·042 to 0·941 and from 0·042 to 0·917, respectively. The loci described were used to cross‐amplify other gobiid species belonging to Gobius, Zosterisessor, Lesueurigobius and Aphia.     

Primers for 10 polymorphic microsatellites from Caulanthus amplexicaulis var. barbarae,and cross‐amplification in other species within the Streptanthoid Complex (Brassicaceae)

Ten microsatellite primer pairs, developed from Caulanthus amplexicaulis var. barbarae (J. Howell) Munz, specifically amplified target loci across a diverse range of taxa from the Streptanthoid Complex, Brassicaceae. All primer pairs produced amplification products from at least 22 of the 23 accessions tested. Product size variation and observable heterozygosity were consistent with what would be expected from the amplification of polymorphic microsatellites across the complex. Our findings demonstrate that microsatellites developed for C. amplexicaulis var. barbarae will be useful for population‐genetic studies in taxa throughout the Streptanthoid Complex.     

Additional microsatellites for Sparus aurata and cross‐species amplification within the Sparidae family

Six new microsatellite loci were isolated and characterized in 32 individuals from a farm population of gilthead seabream (Sparus aurata). Expected heterozygosity at all loci was high, ranging from 0.835 to 0.958 with between 10 and 27 alleles per locus. A multiplex polymerase chain reaction protocol was developed using four of the loci. Cross‐species amplification of the loci was tested in six species of the Sparidae family and four loci were successfully amplified in two or more related species.     

Genetic diversity and population structure of silver pomfret (Pampus argenteus) in the Indo-West Pacific revealed by mitochondrial control region sequences

Sequence variation of the mitochondrial control region in silver pomfret (Pampus argenteus) among six populations from the Indo-West Pacific region was analyzed. A total of 309 polymorphic sites were detected in the 464–466 bp segment of the control region among 165 individuals which defined 72 haplotypes. Haplotype diversity was ranged from 0.800 to 0.943 and nucleotide diversity ranged from 0.333% to 1.886%. Genetic distance and the fixation index indicated significant differences among populations in different geographical groups (P < 0.05). Median-joining network analysis data showed distinct patterns of phylogeographic structure. The present results suggested that P. argenteus populations have developed significant genetic structure, especially among geographically isolated groups.     

Isolation and characterization of novel microsatellite loci for the short‐beaked common dolphin (Delphinus delphis) and cross‐amplification in other cetacean species

Seven tetranucleotide and three dinucleotide microsatellite loci were isolated from the short‐beaked common dolphin (Delphinus delphis). Number of alleles and observed heterozygosity ranged from three to twelve (mean 8.4) and from 0.087 to 0.978 (mean 0.741), respectively. All loci were tested in four other odontocete species and showed polymorphisms in most locus–species combinations. This panel of nuclear markers will be useful for the investigation of population ecology and social interactions in common dolphins and other cetacean species.     

Isolation and characterization of polymorphic microsatellites isolated from the spotless starling (Sturnus unicolor) and cross‐species amplification in the European starling (Sturnus vulgaris)

Spotless and European starlings (Sturnus unicolor and Sturnus vulgaris) have attracted attention from researchers interested in sexual selection, evolution of parental care and reproductive strategies. Both species show high levels of intraspecific nest parasitism and extra‐pair paternity, but research in this area is hampered by a lack of molecular markers specific for these species. Here, we describe a set of primers for nine microsatellite loci in spotless starlings, eight of which are highly polymorphic (7.37 alleles in average). These microsatellites are also polymorphic in European starlings (6.75 alleles in average).     

Isolation and characterization of microsatellites in Leuciscus cephalus (Cypriniformes,Cyprinidae) and cross‐species amplification within the family Cyprinidae

Thirteen polymorphic microsatellites were isolated from Leuciscus cephalus, a widespread cyprinid species with great ecological tolerance. Together with the cross‐species amplification of six additional loci originally published for three cyprinid fish species, we optimized a multiplex panel for L. cephalus allowing the genotyping of 19 polymorphic loci. Number of alleles and heterozygosity per locus in a sample of 20 fish individuals ranged from two to 16 and from 0.05 to 0.90, respectively. These primers will be useful in determining the population structure of L. cephalus. In addition, successful cross‐amplification was obtained for several species of Cyprinidae.     

Development of microsatellite markers for southern catfish (Silurus meriaionalis) and cross‐species amplification in northern sheatfish (Silurus soldatovi)

Forty microsatellite markers were developed from a (CA)_n enrichment library created from the DNA of southern catfish (Silurus meriaionalis). Also, the population structure of northern sheatfish (Silurus soldatovi) was examined by 24 microsatellite loci. They are polymorphic in at least one of the two geographically distant populations sampled from the Heilongjiang River and the Songhuajiang River in North China, respectively. Unbiased expected heterozygosity levels varied from 0.435 to 0.946 and number of alleles per locus varied from three to 20. Results indicated that these microsatellite loci were highly polymorphic and could be used as genetic markers.     

Development and cross‐species amplification of 18 microsatellite markers in the Sumatran tiger (Panthera tigris sumatrae)

Eighteen polymorphic microsatellite loci from the highly endangered Sumatran tiger (Panthera tigris sumatrae) were isolated and characterized. Upon polymerase chain reaction amplification, 16 of these markers produced a single, sharp band in all three tiger and 10 non‐tiger felid species examined. Of the two remaining loci, 6HDZ057 and 6HDZ635 failed to amplify genomic DNA from puma (Felis concolor) and cheetah (Acinonyx jubatus), respectively. The amplification of these markers across four genera is an indication of their usefulness for population genetics studies and conservation work in a wide range of felid species.     

Isolation and characterization of microsatellites in Neogobius kessleri (Perciformes,Gobiidae) and cross‐species amplification within the family Gobiidae

Fourteen polymorphic microsatellites were isolated from Neogobius kessleri, a benthic fish of Ponto‐Caspian origin which has been recently introduced into the Middle and Upper Danube River. Number of alleles and heterozygosity per locus in a sample of 32 fish individuals ranged from two to four and from 0.13 to 0.75, respectively. These primers will be useful in determining the population structure of N. kessleri. In addition, successful cross‐amplification was obtained for four related species, N. melanostomus, N. fluviatilis, N. gymnotrachelus and Proterorhinus marmoratus. These microsatellite loci may be useful for the evaluation of the origin of non‐native goby populations.     

Characterization of microsatellite loci in silver carp (Hypophthalmichthys molitrix), and cross‐amplification in other cyprinid species

Captive populations of silver carp (Hypophthalmichthys molitrix), a major aquaculture species in Asia, would undoubtedly benefit from genetic monitoring and improvement programs. We report the isolation and preliminary characterization of 16 microsatellite loci derived from both conventional and microsatellite‐enriched libraries. Inheritance studies confirmed the allelic nature of observed polymorphisms at all loci, while identifying null alleles at two loci. These loci, having varying degrees of polymorphism, should provide useful markers for applied genetic studies. A high degree of cross‐amplification among 10 other cyprinid species suggests that these loci may have more widespread utility.     

Isolation and characterization of polymorphic microsatellites in Labeo rohita and their cross‐species amplification in related species

The major Indian carps namely rohu (Labeo rohita), catla (Catla catla), mrigal (Cirrhinus mrigala) and calbasu (Labeo calbasu) are important freshwater species of the Indian subcontinent constituting over 65% of the fish produce. In the present study, isolation of 12 microsatellite loci from rohu has been reported. Cross‐species amplification in related carps and their implication in population genetic studies as well as selective breeding program were discussed.     

Isolation and cross‐familial amplification of 41 microsatellites for the brook charr (Salvelinus fontinalis)

The brook charr (Salvelinus fontinalis; Osteichthyes: Salmonidae) is a phenotypically diverse fish species inhabiting much of North America. But relatively few genetic diagnostic resources are available for this fish species. We isolated 41 microsatellites from S. fontinalis polymorphic in one or more species of salmonid fish. Thirty‐seven were polymorphic in brook charr, 15 in the congener Arctic charr (Salvelinus alpinus) and 14 in the lake charr (Salvelinus namaycush). Polymorphism was also relatively high in Oncorhynchus, where 21 loci were polymorphic in rainbow trout (Oncorhynchus mykiss) and 16 in cutthroat trout (Oncorhynchus clarkii) but only seven and four microsatellite loci were polymorphic in the more distantly related lake whitefish (Coregonus clupeaformis) and Atlantic salmon (Salmo salar), respectively. One duplicated locus (Sfo228Lav) was polymorphic at both duplicates in S. fontinalis.     

Polymorphic microsatellite loci from the western corn rootworm (Insecta: Coleoptera: Chrysomelidae) and cross‐amplification with other Diabrotica spp

Corn rootworms (Diabrotica spp.) make up the major insect pest complex of corn in the US and Europe, and there is a need for molecular markers for genetics studies. We used an enrichment strategy to develop microsatellite markers from the western corn rootworm (Diabrotica virgifera virgifera). Of 54 loci isolated, 25 were polymorphic, and of these, 17 were surveyed for variability in 59 wild individuals. In addition, the potential for cross‐amplification of these microsatellites was surveyed for Mexican, northern, and southern corn rootworms. Nine microsatellite loci showed Mendelian inheritance and are likely to be useful in population genetics studies.     

Genetic diversity and differentiation of 12 eastern Adriatic and western Dinaric native sheep breeds using microsatellites

Nuclear genetic diversity and differentiation of 341 sheep belonging to 12 sheep breeds from Croatia and Bosnia and Herzegovina were examined. The aim of the study was to provide the understanding of the genetic structure and variability of the analysed pramenka sheep populations, and to give indications for conservation strategies based on the population diversity and structure information. The genetic variation of the sheep populations, examined at the nuclear level using 27 microsatellite loci, revealed considerable levels of genetic diversity, similar to the diversity found in other European indigenous low-production sheep breeds. Population-specific alleles were detected at most loci and in breeds analysed. The observed heterozygosity ranged from 0.643 (in Lika pramenka) to 0.743 (in Vlasic pramenka), and the expected heterozygosity ranged from 0.646 (in Lika pramenka) to 0.756 (in Dalmatian pramenka). Significant inbreeding coefficients were found for half of the populations studied and ranged from 0.040 (Pag island sheep) to 0.091 (Kupres pramenka). Moderate genetic differentiation was found between the studied sheep populations. The total genetic variability observed between different populations was 5.29%, whereas 94.71% of the variation was found within populations. Cres island sheep, Lika pramenka and Istrian sheep were identified as the most distinct populations, which was confirmed by the factorial analysis of correspondence and supported through a bootstrapping adjustment to correct for the difference in the sample sizes. The population structure analysis distinguished 12 clusters for the 12 sheep breeds analysed. However, the cluster differentiation was low for Dalmatian, Vlasic, Stolac and Krk pramenka. This systematic study identified Lika pramenka and Rab island sheep as those with the lowest diversity, whereas Istrian sheep and Pag island sheep had the highest. Conservation actions are proposed for Istrian, Rab and Cres island sheep, Lika and Kupres pramenka because of high estimated coefficients of inbreeding.     

Microsatellite DNA markers for American shad (Alosa sapidissima) and cross‐species amplification within the family Clupeidae

Sixteen microsatellite loci were identified and characterized for American shad (Alosa sapidissima). The number of alleles per locus observed ranged from eight to 32 and averaged 15.4 alleles. Average observed heterozygosity was 81.1%. The markers were screened using four other species from the family Clupeidae. Amplification success among Alosa species was 79.2% with 81.6% polymorphism among those markers that amplified successfully. Amplification success was poor in Dorosoma (31.3%). Due to allelic diversity and estimates of heterozygosity, these markers can be useful in A. sapidissima for population level analyses, parentage assignment and broodstock management.     

利用碳氮稳定同位素技术分析东海银鲳食性

利用稳定同位素技术分析了东海银鲳(Pampus argenteus)及其可能摄食饵料的碳、氮稳定同位素比值,以期探讨东海银鲳可能的食物来源.结果表明:东海区银鲳平均碳(813C)和氮(δ15N)稳定同位素比值分别为-18.22‰、8.16‰,其可能摄食饵料的δ13C值变化范围为-17.33‰～-21.58‰,差值为4.25‰,δ15N值变化范围为3.89‰～7.96‰,差值为4.07‰;东海银鲳可能的食物来源主要为箭虫、虾类、水母类、头足类、仔稚鱼和浮游动物等,其中箭虫为主要可能的食物来源,其贡献率为24%～78%,平均贡献率为57%;银鲳可能摄食的其他饵料中,贡献率从大到小依次为虾类、水母类、头足类、仔稚鱼、＞1000 μm浮游动物、500～1000 μm浮游动物和100～500 μm浮游动物,其平均贡献率分别为11.8%、8.4%、7.1%、5.0%、4.9%、3.2%和2.6%.由以上结果可知,银鲳是一种广食性鱼类,其饵料种类较多,碳和氮的来源均较为复杂.