Molecular characterization of Fasciola spp. from the endemic area of northern Iran based on nuclear ribosomal DNA sequences

Fasciolosis caused by Fasciola spp. (Platyhelminthes: Trematoda: Digenea) is considered as the most important helminth infection of ruminants in tropical countries, causing considerable socioeconomic problems. In the endemic regions of the North of Iran, Fasciola hepatica and Fasciola gigantica have been previously characterized on the basis of morphometric differences, but the use of molecular markers is necessary to distinguish exactly between species and intermediate forms. Samples from buffaloes and goats from different localities of northern Iran were identified morphologically and then genetically characterized by sequences of the first (ITS-1) and second (ITS-2) Internal Transcribed Spacers (ITS) of nuclear ribosomal DNA (rDNA). Comparison of the ITS of the northern Iranian samples with sequences of Fasciola spp. from GenBank showed that the examined specimens had sequences identical to those of the most frequent haplotypes of F. hepatica (n = 25, 48.1%) and F. gigantica (n = 20, 38.45%), which differed from each other in different variable nucleotide positions of ITS region sequences, and their intermediate forms (n = 7, 13.45%), which had nucleotides overlapped between the two Fasciola species in all the positions. The ITS sequences from populations of Fasciola isolates in buffaloes and goats had experienced introgression/hybridization as previously reported in isolates from other ruminants and humans. Based on ITS-1 and ITS-2 sequences, flukes are scattered in pure F. hepatica, F. gigantica and intermediate Fasciola clades, revealing that multiple genotypes of Fasciola are able to infect goats and buffaloes in North of Iran. Furthermore, the phylogenetic trees based upon the ITS-1 and ITS-2 sequences showed a close relationship of the Iranian samples with isolates of F. hepatica and F. gigantica from different localities of Africa and Asia. In the present study, the intergenic transcribed spacers ITS-1 and ITS-2 showed to be reliable approaches for the genetic differentiation of Fasciola spp., providing bases for further studies on F. hepatica, F. gigantica and their intermediate forms in the endemic areas in Asia.     

First report of the invasive eel pest Pseudodactylogyrus bini in North America and in wild American eels

We detected 2 species of monogenean gill worms, Pseudodactylogyrus bini (Kikuchi, 1929) Gusev, 1965 and P. anguillae (Yin & Sproston, 1948) Gusev, 1965 (Monopisthocotylea: Pseudodactylogyridae), on American eel Anguilla rostrata in 2 rivers in South Carolina, USA. One of these, P. anguillae, was reported 5 yr ago from Nova Scotia; as well as in South Carolina, we also discovered it in 2 localities in Chesapeake Bay. Differences in the morphologies of specimens of either species of worm from North America and northeastern Asia were negligible. Similarly, the level of variation in sequences in the ITS2 (internal transcribed spacers) region of ribosomal RNA was minor, and not consistent with geographical origin. These data indicate that these monogeneans invaded North America only recently, possibly in parallel with the nematode Anguillicola crassus (which is known to have been introduced with commercial imports of foreign eels). We map the current global occurrence of these monogeneans, and conclude that their dispersal from northeastern Asia was largely as a result of the eel trade, and has probably been secondarily augmented by longshore migration of infected eels, and possibly also by transport in ballast waters. With present technology, all eel stocks must still be collected from the wild; unless shipments are disinfected at quarantine, these and other eel pathogens (such as A. crassus) are likely to continue to colonise other regions of the world.     

Genetic biogeography of the rate “copper moss”,Scopelophila cataractae (Pottiaceae)

Scopelophila cataractae, one of the so-called copper mosses, has a broad geographic distribution that includes North, Central, and South America, Europe, and Asia, but is rare throughout its range. A genetic analysis of 32 populations from the United States, Europe, and Asia based on 15 putative allozyme loci indicates that levels of genetic diversity vary among geographic regions. Six European populations are fixed for the same alleles at all 15 loci, consistent with the hypothesis thatS. cataractae is a recent immigrant in that region. The species is more diverse in the U.S., where it appears to be native. Five populations collected on copper-enriched soils around shrines and temples in Tokyo are genetically monomorphic, but Asian populations from another Japanese site, India, and Nepal are exceptionally diverse in terms of numbers of alleles and multilocus haplotypes, total gene diversity (H_T), and in the degree of differentiation among populations (measured as Nei'sI andD). Long-distance dispersal has probably played an important role in the geographic history ofS. cataractae, but the species appears to be native in both the New and Old Worlds. Gene flow between plants disjunct on different continents is insufficient to explain the lack of geographically correlated morphological and genetic differentiation inS. cataractae.     

Molecular genetic structure and evolution in native and colonized populations of the Chinese mitten crab, Eriocheir sinensis

Chinese mitten crab (Eriocheir sinensis), a native species in China, has populated Continental Europe and North America since 1912. In this paper, the nucleotide variation in the fragments of mitochondrial COII (693 bp), Cytb (766 bp), and nucleotide ITS (706 bp) was analyzed in native (Yantgze, Yellow, and Liaohe Rivers in China) and colonized (Elbe, Rhine, and Thames Rivers in Europe, and San Francisco Bay in North America) populations of the Chinese mitten crab. The major findings are as follows. First, the genetic variability in the native populations is higher than that in the colonized European and North American populations, with the exception of the Elbe River population, which possesses a similar level of variability with the native populations. Second, a remarkable loss of singletons has been associated with the colonization of Chinese mitten crabs. Third, the AMOVA and F _ST results demonstrate that there are no significant genetic differentiations among the populations from the three continents, but there is a significant differentiation between pairwise populations within and among continents. Fourth, it is found that expansion-drift and gene flow pattern are involved in the European populations. The neutrality test and R ₂ statistics suggest that a moderate founder population exists in the colonized populations, and only the Yangtze River population has undergone a recent population expansion. Finally, the results demonstrate that the European populations originate from multiple rivers in China on multiple occasions. The San Francisco population originates from both the native Chinese populations and the colonized European populations, most likely the Thames population.     

Decline in non-native freshwater eels in Japan: ecology and future perspectives

The occurrence, distribution, and biological characteristics of non-native freshwater eels were analyzed using 5524 eels collected from 16 sites in Japan between 1997 and 2005. Three hundred seventy-four fishes (6.8%) were identified as non-native European eels, Anguilla anguilla, while the remainder (93.2%) were native Japanese eels, A. japonica. The European eel was found at 7 sites (44%), including 3 rivers, 2 freshwater lakes, one brackish lake, and one sea bay, suggesting a wide rage of habitat use. This variability of habitat use was also evidenced by the otolith microchemistry, which showed that they had lived in not only freshwater but also in seawater habitats. The sites with European eel were localized within the vicinity of southern Japan where a number of these eels were cultivated in the early 1970’s, suggesting that some had escaped from the culture ponds or were released intentionally into nearby natural waters. The large body size (mean total length: 803 mm), pigmented skin, enlarged eyes, and relatively matured gonads (mean gonad somatic index: 1.9) found in non-native European eels indicated that most had metamorphosed into the migratory silver phase, suggesting their ability to initiate spawning migration. However, the proportion of European eels in Mikawa Bay in 1997 was more than 12%, which decreased markedly to less than 2% after 2001, corresponding to the recent decline in import of European glass eels for aquaculture. This suggests that the population of European eels will decrease in Japanese waters in the future.     

Validation of the occurrence of the American eel Anguilla rostrata (Le Sueur, 1817) in free-draining European inland waters

The screening of 2,735 eels from European waters and aquaculture farms was conducted using mitochondrial Cytochrome b and 16S rRNA gene fragments amplified by polymerase chain reaction. Reaction products were either sequenced directly or subjected to analysis using restriction fragment length polymorphism which resulted in species-specific restriction patterns. Beside the expected European eel, Anguilla anguilla (Linnaeus, 1758), the American eel, Anguilla rostrata (Le Sueur, 1817), was also identified in samples from both aquaculture (N = 40 out of 1,025) and from natural waters (N = 44 out of 1,710). The life stages of American eels identified from several German waters draining to either the Baltic Sea and the North Sea ranged from elver to silver eels. This indicates that stocking with glass eels or elvers must have occurred several times most likely in the period from 1998 to 2002. The application of a fast and precise method for species identification and genetic monitoring of eels delivered for stocking is therefore essential for maintaining the autochthonous species composition in future. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Natural Enemies ofTrapaspp. in Northeast Asia and Europe

Trapa natansis an aquatic plant native to the Old World. In the late 1880s this plant was found to be naturalized in the eastern United States where it has become an important weed. To evaluate the potential of biological control to reduce this weed, surveys for natural enemies ofTrapaspp. were made in Northeast Asia and Europe. In addition, populations ofTrapa japonicain South Korea were monitored for one season to evaluate the occurrence and impact of insect natural enemies. Among the insects found, the leaf beetleGalerucella birmanicawas the most common and damaging species in Asia. It can cause complete defoliation of whole populations of plants. Nymphuline pyralid moths were also common and at times damaging. Both the beetle and the moths feed and develop on unrelated plants and so have no potential as biological control agents ofT. natansin North America. TwoNanophyesweevils were found in Asia which feed in petiole floats of the leaves. They are thought to be specific toTrapabut were not observed to be damaging. Low level populations of polyphagous Homoptera were common. Chironomid midges were also frequently associated with the plants, but for the most part were filter feeders, not herbivores. The greatest impact on the monitored plants in South Korea was by the leaf beetleG. birmanicawhich defoliated most of the plants, causing the mats of plants to sink. In Europe, a similar and related insect fauna was found, but none of the insects was very damaging to the plants. One Italian weevil,Bagous rufimanus,feeds within the fruit stems and might be more damaging at higher than observed population levels. Some diseases were seen on some plants in both regions. European populations ofT. natanshave declined significantly, stimulating conservation efforts. Literature records ofTrapainsects from warm areas are presented. These could be of interest ifT. natansspreads into warmer areas of North America.     

Long-term effects of hydropower installations and associated river regulation on River Shannon eel populations: mitigation and management

The Shannon, Ireland’s largest river, is used for hydroelectricity generation since 1929. Subsequently, the Electricity Supply Board assumed responsibility for management of its eel stocks, due to the impact of the hydro-dam on recruitment to the commercial fishery. In order to negate a decline in juvenile recruitment resulting from the installation of hydroelectric facilities, management was focused on stocking lakes with elvers and fingerling eels. These were trapped at the hydropower facilities and in estuarine tributaries during their up-stream migrations. Due to the decline of natural recruitment in more recent times, attempts have also been made to develop an estuarine glass eel fishery. Stock levels are then monitored through annual surveys of the population trends of juvenile (glass eel, elver), growing phase (yellow eel) and downstream migrating pre-spawners (silver eels). Survey results and fishery management programmes are reviewed in this article. In addition to the long-term effects the hydroelectric facilities have had on the stock levels, there is also an annual effect on the migratory patterns of downstream migratory silver eels. In the lower reaches of the river system flow rates are regulated by the hydroelectric stations. We review previous work that had highlighted the importance of flow in determining the timing of the silver eels migrations, and assess the relationship between flow and migration in more detail through the use of hydroacoustic and telemetric studies. Current research on seaward migrating silver eel populations, suggests that spawner escapement rates can most effectively be increased by trapping migrating eels at fishing weirs located up-stream of the power station and transporting them towards the estuary. Guest editors: R. L. Welcomme & G. Marmulla Hydropower, Flood Control and Water Abstraction: Implications for Fish and Fisheries     

Hyperparasitism by Mesochorus spp. (Hymenoptera: Ichneumonidae) in Peristenus sp. (Hymenoptera: Braconidae) and development of PCR primers for hyperparasitoid detection

Peristenus sp. pupae collected from Lygus spp. nymphs in 2001 and 2002 were over-wintered in the laboratory. In both years, more than 30% of adults emerging from over-wintering pupae were identified as ichneumonid hyperparasitoids, Mesochorus curvulus Thomson and Meschorus sp. (Hymenoptera: Ichneumonidae). At the end of the over-wintering period, Peristenus sp. males emerged first followed by Peristenus sp. females and finally Mesochorus spp. The male:female ratio in emerging Peristenus sp. adults was skewed towards males. The Internal Transcribed Spacer (ITS) region and the cytochrome oxidase I (COI) gene from Mesochorus spp. were sequenced. ITS sequences were used to develop PCR primers to detect Mesochorus spp. hyperparasitism in the primary host, Lygus spp. PCR analysis of field-collected Lygus spp. nymphs gave similar estimates of Mesochorus spp. hyperparasitism to the rearing protocols (25–28%). Sequence analysis of COI and ITS regions and subsequent restriction endonuclease analysis of ITS PCR products from Mesochorus spp. indicate the presence of two genotypes in the population. The possibility that these two genotypes represent separate or cyrptic species is discussed.     

我国口岸截获芒果象属检疫性有害生物疫情分析

【目的】芒果象属昆虫是我国禁止进境的植物检疫性有害生物,广泛分布于非洲、北美洲和东南亚,寄主为芒果。其生活隐蔽,羽化孔未出现时看不出危害状,危害率可达30%~80%,严重影响芒果的产量和质量。近年来,我国口岸截获的芒果象甲的数量日益增多,表明该类检疫性有害生物传入我国的风险越来越大。【方法】运用Microsoft Excel工作表对2003—2015年全国口岸截获芒果象属有害生物的疫情进行统计分析。【结果】2003—2015年全国口岸共截获芒果象4156批次,22个直属局有截获记录。其中,截获芒果果核象甲3028批次,数量最多,占截获芒果象总批次的72.86%;芒果果实象甲和芒果果肉象甲相对较少,分别为837和291批次。截获芒果象的来源国主要为非洲和东南亚芒果象疫情分布国家和地区,且主要自进境旅客随身携带物中检出。【结论】我国口岸应加大对芒果象的检疫力度,保护我国农林业生产安全。     

Phylogeny and taxonomy of powdery mildew fungi of Erysiphe sect. Uncinula on Carpinus species

A phylogenetic analysis of the Erysiphe with uncinuloid ascoma appendages (Erysiphe section Uncinula, Erysiphales, Ascomycota) on Carpinus spp. was done using sequences of the rDNA ITS regions and the D1/D2 domains of the 28S rDNA. These results, combined with morphological data, revealed a complex consisting of several distinct taxa. These included the already described Erysiphe carpinicola on C. japonica distinguishable from the Erysiphe sp. on C. betulus and C. tschonoskii as well as the one on C. laxiflora. Thus, it was shown that Oidium carpini, described from Europe on Carpinus betulus, the powdery mildew with uncinula-like ascomata, recently found in Europe on this host, as well as an Erysiphe on C. tschonoskii in Japan, described previously as E. carpinicola, all belong to a single new species, named E. arcuata in this paper. As the powdery mildew on C. laxiflora was also distinct from other known species, it is named E. carpini-laxiflorae in this paper. The already described E. pseudocarpinicola and Erysiphe sp. on Carpinus cordata are two additional taxa, which are morphologically and genetically distinguished from the other species of Erysiphe sect. Uncinula on Carpinus spp.     

Re-consideration of Peronospora farinosa infecting Spinacia oleracea as distinct species, Peronospora effusa

Downy mildew is probably the most widespread and potentially destructive global disease of spinach (Spinacia oleracea). The causal agent of downy mildew disease on various plants of Chenopodiaceae, including spinach, is regarded as a single species, Peronospora farinosa. In the present study, the ITS rDNA sequence and morphological data demonstrated that P. farinosa from S. oleracea is distinct from downy mildew of other chenopodiaceous hosts. Fifty-eight spinach specimens were collected or loaned from 17 countries of Asia, Europe, Oceania, North and South America, which all formed a distinct monophyletic group. No intercontinental genetic variation of the ITS rDNA within Peronospora accessions causing spinach downy mildew disease was found. Phylogenetic trees supported recognition of Peronospora from spinach as a separate species. Microscopic examination also revealed morphological differences between Peronospora specimens from Spinacia and P. farinosa s. lat. specimens from Atriplex, Bassia, Beta, and Chenopodium. Consequently, the name Peronospora effusa should be reinstated for the downy mildew fungus found on spinach. Here, a specimen of the original collections of Peronospora effusa is designated as lectotype.     

Swimming performance of silver eels is severely impaired by the swim-bladder parasite Anguillicola crassus

Infection with the swim-bladder parasite Anguillicola crassus is suggested as one of the principal causes of the collapse of the European eel population. This nematode has been introduced in Europe from Asia in the 80s and parasitized in a short time Anguilla eel species in different geographical regions across the globe. The parasites drain energy due to their sanguivorous feeding and they cause mechanical damage on the swim-bladder wall. These two effects are hypothesized to impair the spawning migration of the European eel. In this study, we have investigated both effects on swimming performance. We hypothesized that parasitic sanguivorous activities - related to parasite weight - reduce swimming endurance, while mechanical damage of the swim-bladder impairs buoyancy control. Eighty eels suffering various degrees of infection were introduced in swim-tunnels and subjected to a swimming fitness test. The relation between A. crassus infection and swimming efficiency was measured for large female silver eels swimming at various speeds. Infected eels had lower cruising speeds and a higher cost of transport. Eels without parasites, but with a damaged swim-bladder showed similar effects. Almost half of the eels that contained damaged swim-bladders (43%) stopped swimming at low aerobic swimming speeds (< 0.7 m/s). Simulated migration trials in a recent related study have confirmed that eels with a high parasite level or with damaged swim-bladder show early migration failure (< 1000-km). Reduced swimming performance appears to be associated with swim-bladder dysfunction. As we found that especially silver eels have much higher infection levels than yellow eels, it is concluded that migrating silver eels with severely infected or damaged swim-bladders are unable to reach the spawning grounds.     

Stable isotope analyses provide new insights into ecological plasticity in a mixohaline population of European eel

Recent studies have shown that anguillid eel populations in habitats spanning the marine–freshwater ecotone can display extreme plasticity in the range of catadromy expressed by individual fishes. Carbon and nitrogen stable isotope analysis was used to differentiate between European eels (Anguilla anguilla) collected along a short (2 km) salinity gradient ranging from <1‰ to ~30‰ in Lough Ahalia, a tidal Atlantic lake system. Significant differences were recorded in mean δ¹³C, δ¹⁵N and C:N values from eels collected from fresh, brackish and marine-dominated basins. A discriminant analysis using these predictor variables correctly classified ca. 85% of eels to salinity zone, allowing eels to be classified as freshwater (FW), brackish (BW) or marine (MW) residents. The results of the discriminant analysis also suggested that a significant proportion of eels moved between habitats (especially between FW and BW). Comparisons of several key population parameters showed significant variation between eels resident in different salinity zones. Mean condition and estimated age was significantly lower in MW eels, whilst observed length at age (a correlate of growth) was significantly higher in MW eels, intermediate in BW and lowest in FW eels. This study has demonstrated that the ecology of eels found along a short salinity gradient can be extremely plastic and that stable isotope analysis has considerable utility in demonstrating intra-population variation in diadromous fishes.     

Subdivision of the genus Pleuroxus (Anomopoda,Chydoridae) into subgenera worldwide

Examination of many species of Pleuroxus from the entire World has revealed a major dichotomy in morphology, one line related to aduncus: the other to laevis both from Europe. Pleuroxus trigonellus (the type species of the genus) and Peracantha truncata are in two other distinct lines. The new subgenus Tylopleuroxus is proposed for the species related to aduncus, and the subgenus Picripleuroxus for those related to laevis. Tylopleuroxus is primarily Southern Hemisphere in distribution, Picripleuroxus primarily Northern Hemisphere. Evidence thus far suggests that probably most, perhaps all, of the taxa are confined to single continents or land masses. Deceased     

基于ITS序列分析钩苞大丁草九个居群的亲缘关系

该研究对我国西南地区钩苞大丁草(Gerbera delavayi)9个居群rDNA ITS序列进行PCR的扩增和检测序列,并以非洲菊(G.jamesonii)的ITS序列作为外类群,比较了序列之间的差异,同时分析了钩苞大丁草不同居群在地理距离与遗传距离之间的关系,构建了NJ系统发育树。结果表明:(1)钩苞大丁草9个居群的ITS序列全长介于600~700 bp之间,平均长度约为657 bp,其中,ITS1长度为243~246 bp,(G+C)含量为45.67%~46.80%之间,5.8S长度191~193 bp,(G+C)含量为58.60%~58.61%之间,ITS2长度为220~221 bp,(G+C)含量为57.00%~57.45%之间;ITS序列共有22个变异位点,ITS1序列(17个)、5.8S序列(2个)以及ITS2序列(3个)上均有变异。(2)地理距离与遗传距离有正相关(r2=0.652),序列间遗传分化距离为0.001 1~0.024 3,其中普洱居群与其他居群间遗传距离最大。(3)钩苞大丁草9个居群分成三个分支,普洱居群单独成支,丽江和洱源居群聚为一支,富源、武定、德昌、石林、新平和开远6个居群聚为一支。rDNA ITS序列可以用于钩苞大丁草群体遗传研究的分析,该研究结果为其保护性开发提供了参考依据。     

Annual variability in upstream migration of glass eels in a southern USA coastal watershed

We investigated the environmental factors that affected temporal variability of eel recruitment and upstream migration in a freshwater coastal river along the southeastern US. Glass eels Anguilla rostrata were collected through ichthyoplankton sampling in the lower Roanoke River, North Carolina. Monthly samples were taken from fixed stations from May 2001 through June 2003. There was no evidence of consistent seasonal migration patterns for glass eels in Roanoke River. From May through December in 2001, glass eels were captured only during August. In 2002, glass eels arrived in February and remained in ichthyoplankton samples through October, with the exception of samples from September. Peak catch occurred in March at 4.02 ± 1.2 and declined through June to 0.18 ± 0.07 (#/1,000 m³). By August, the mean density increased to 0.96 ± 0.82 and to 3.59 ± 2.77 by October. In 2003 from January through June, glass eels were captured only during February and March. Glass eels were routinely collected when river discharge rates were <150 m³ s⁻¹. River discharge rates >650 m⁻³ s⁻¹ resulted in no glass eels in our samples. Upstream migration during 2002 was not correlated with water temperature or related to lunar phase. Glass eel freshwater upstream migration was initiated when water temperatures exceeded a threshold range of 10°C to 15°C; however, glass eels continued to migrate when water temperatures approached 30°C. The overall negative effect of river discharge suggests that changes in the water release schedules of upstream hydroelectric facilities during glass eel migration could strongly influence their recruitment success.     

Genetic diversity in the rare orchid Gymnadenia odoratissima and a comparison with the more common congener, G. conopsea

Patterns and levels of genetic diversity mayhave significant influence on the long termpersistence of local populations and revealingsuch information is important in protectingrare species. In this study we investigated thegenetic pattern in five microsatellite lociwithin five Swedish populations of the rareorchid species Gymnadenia odoratissima. Thegeographic distribution of G. odoratissima isrestricted to Europe and in Scandinavia it isonly found in three provinces in southernSweden; Östergötland,Västergötland and on the island ofGotland.Compared with the more widespread congener G.conopsea our results indicate lower levels ofgenetic variation within and higher degrees ofgenetic differentiation among populations ofG. odoratissima (H_EL = 0.6–0.8 in G. conopseaand 0.3–0.7 in G. odoratissima; F_ST over allpopulations = 0.06 in G. conopsea and 0.19 inG. odoratissima). Also, we found a cleardistinction among mainland and islandpopulations of G. odoratissima wherepopulations on the island of Gotland seem toexhibit higher levels of gene flow andintragenetic variation, probably as a result ofa larger number of existing populations.Future conservation of this species shouldfocus on facilitation on colonisation events,especially on the mainland, and preservation ofthe genetically more variable Gotlandpopulations.     

Water temperature and upstream migration of glass eels in New Zealand: implications of climate change

Glass eels migrating upstream in a New Zealand river showed a clear preference for water temperatures between 12 and 20°C, with an optimum of 16.5°C. Water temperatures <12°C and >22°C almost completely inhibited migration, which implies that warmer temperatures associated with global climate change might have a detrimental impact on glass eel recruitment in their current ranges. We established this by trapping glass eels of shortfin, Anguilla australis, and longfin, A. dieffenbachii, eels nightly from September to November. Eels caught in 2001 (50,287) outnumbered those caught in 2002 (19,954); shortfin glass eels dominated catches in both years, comprising 91–93% of the catch. Longfins were larger than shortfins, and size and pigmentation in both species increased as the seasons progressed. Temperatures within the migratory season in 2001 showed ∼14-day intervals between maxima that appeared to be associated with the new and full moons.