Freshwater Subspecies Research Articles

Morphology of sweat glands in the skin of Saimaa (Pusa hispida saimensis) and Baltic ringed (Pusa hispida botnica) seals.

The endangered Saimaa ringed seal (Pusa hispida saimensis) is an endemic freshwater subspecies inhabiting Lake Saimaa in Finland. The Baltic ringed seal (Pusa hispida botnica) inhabits the brackish Baltic Sea, which is almost entirely landlocked. Recent research shows that Saimaa and Baltic ringed seals may be genetically even further apart from each other than from other ringed seal subspecies. We documented histologically the integument microstructure of Saimaa and Baltic ringed seals to determine whether the geographic and genetic isolation was manifested as variation in the integument microstructure of these subspecies adapted to icy aquatic environments. The skin structures of these subspecies were similar to those of other phocids. The association of the sweat glands with hair follicles in both subspecies suggested that they were small apocrine sweat glands described previously in terrestrial or aquatic mammals. None of the apocrine glands had large lumina, and some of the ducts were relatively straight and short. Further studies analysing the mode of secretion, for example, apocrine versus eccrine, in the sweat glands are necessary to confirm the types of sweat glands in seals.

Comparative Analysis of Blood Transcriptome in the Yangtze Finless Porpoise (Neophocaena asiaeorientalis)

The Yangtze finless porpoise (Neophocaena asiaeorientalis) is the sole freshwater subspecies of Neophocaenaphocaenoides, and there is a lack of data on its transcriptome. In this study, we applied RNA-seq technology to assemble, de novo, a transcriptome and analyzed differential expressed genes (DEGs). About 6 Gb of clean data was generated for the Yangtze finless porpoise blood (n = 6) through de novo sequencing. In total, 151,211 unigenes were generated and a total of 119,039 of these unigenes (78.72%) were functionally annotated when searched for within the NCBI Nr, SwissProt, GO, COG, and KEGG databases. Diverse and extensive expressed gene catalogs were sampled for the Yangtze finless porpoise. DESeq2 was used to analyze the differential expression genes (DEGs) obtained from the assembled transcriptome. The results indicated that DEGs have close relationships with the Yangtze finless porpoise’s development, evolution and adaptation. Further, we found that genes involved in cetacean TAG synthesis might directly explain the molecular basis of cetacean blubber thickening. These transcriptome data will assist in understanding molecular mechanisms of Yangtze finless porpoise adaptation.

Assembly and characterization of the MHC class I region of the Yangtze finless porpoise (Neophocaena asiaeorientalis asiaeorientalis).

The Yangtze finless porpoise (Neophocaena asiaeorientalis asiaeorientalis; YFP) is the sole freshwater subspecies of N. asiaeorientalis and is now critically endangered. Major histocompatibility complex (MHC) is a family of highly polymorphic genes that play an important immunological role in antigen presentation in the vertebrates. Currently, however, little is known about MHC region in the genome of the YFP, which hampers conservation genetics and evolutionary ecology study using MHC genes. In this work, a nucleotide sequence of 774,811 bp covering the YFP MHC class I region was obtained by screening a YFP bacterial artificial chromosome (BAC) library, followed by sequencing and assembly of positive BAC clones. A total of 45 genes were successfully annotated, of which four were MHC class I genes. There are high similarities among the four YFP MHC class I genes (>94%). Divergence in the coding region of the four YFP MHC class I genes is mainly localized to exons 2 and 3, which encode the antigen-binding sites of MHC class I genes. Additionally, comparison of the MHC structure in YFP to those of cattle, sheep, and pig showed that MHC class I genes are located in genome regions with regard to the conserved genes, and the YFP contains the fewest MHC class I genes among these species. This is the first report characterizing a cetacean MHC class I region and describing its organization, which would be valuable for further investigation of adaptation in natural populations of the YFP and other cetaceans.

Annotations on marine and freshwater leeches (Annelida, Clitellata, Hirudinea) from North Africa

The purpose of this study is to provide information on the leech fauna of North Africa. This overview is based primarily on collected material, data from the literature and material in museum collections. A total of 37 species belonging to 6 families and 19 genera are presently known. Among these species, eight represent new records for North African countries. This report includes the first records for Tunisia of one freshwater leech, Helobdella stagnalis (Linneaus, 1758), and one marine leech, Trachelobdella lubrica (Grube, 1840). Four taxa are reported as new for Algeria: two marine species (Pontobdella muricata Linnaeus, 1758 and Branchellion torpedinis Savigny, 1822) and two freshwater subspecies (Dina punctata punctata Johansson, 1927 and Dina punctata maroccana Nesemann & Neubert, 1994). Pontobdella muricata is reported for the first time from Morocco and Alboglossiphonia iberica Jueg, 2008, previously known only from the Iberian Peninsula, is now reported from Morocco.

Concentrations of osmotically related constituents in plasma and urine of finless porpoise (Neophocaena asiaeorientalis): implications for osmoregulatory strategies for marine mammals living in freshwater

Abstract Background Most cetaceans inhabit the hyperosmotic marine environment with only a few species living in freshwater habitats. The Yangtze finless porpoise (Neophocaena asiaeorientalis asiaeorientalis) is the only freshwater subspecies of the genus. Our aim was to study whether the osmoregulation mechanism of the Yangtze finless porpoise is different from the marine subspecies, the East Asian finless porpoise (Neophocaena asiaeorientalis sunameri). We assayed and compared the concentrations of the constituents involved in osmoregulation in the blood and urine in the Yangtze finless porpoise and the East Asian finless porpoise. We also compared the corresponding urine constituents of the porpoises with existing data on fin whales (Balaenoptera physalus) and bottlenose dolphins (Tursiops truncatus). Results The mean plasma osmolality of Yangtze finless porpoise was significantly lower than that of the marine subspecies (P < 0.01). Similarly, the urine osmolality of Yangtze finless porpoise was also significantly lower than that of its marine counterpart (P < 0.05). However, the urine sodium concentration of freshwater finless porpoise was significantly lower than that in the marine subspecies (P < 0.01), even though their serum sodium has no significant difference. Moreover, the freshwater porpoise has significantly lower urine urea concentration but much higher serum urea than in the marine finless porpoise (P < 0.05). Conclusions These results suggest that the freshwater finless porpoise does have different osmoregulatory mechanism from marine cetaceans. Conserving sodium by excreting urine with low ion levels may be an essential strategy to maintain the serum electrolyte balance for the freshwater subspecies that also appears to be more susceptible to hyponatremia.

Oligochaeta (Annelida) of the profundal of Lake Hazar (Turkey), with description of Potamothrix alatus hazaricus n. ssp..

Lake Hazar is an alkaline oligotrophic lake of tectonic origin, located in the Eastern Anatolia region in Turkey, 1248 a a.s.l. Its surface area is 80 km2, the average depth 93 m and maximum depth 205 m. The lake and its surroundings an under protection as a region of historical value. During the present study (2007-2012), samples were taken from 15 stations located at a depth of 2-200 m. Oligochaeta comprised 69% of the total invertebrate abundance. The profundal olgochaete fauna was found to consist of only three tubificid taxa, all of the subfamily Tubificinae. Potamothrix alatus hazaricus Timm & Arslan, n. ssp. was dominating anywhere down to maximum depths while Psammoryctides barbatus (Grube) and Ilyodrilus(?) sp. occurred seldom. All three are new records for Lake Hazar. Potamothrix alatus hazaricus shares the "winged" body shape in its genital region with the nominal, brackish-water subspecies P. a. alatus Finogenova, 1972, and the lateral position of the spermathecal pores and the shape of the ventral chaetae with the freshwater subspecies P. a. paravanicus Poddubnaja & Pataridze, 1989 known from Transcaucasian lakes. The mitochondrial COI barcoding gene suggests long separation between the two taxa, but the nuclear ITS region shows no variation. The generic position of Ilyodrilus (?) sp. remains obscure since its internal genitalia could not be studied.

Establishment and characterization of fibroblast cell lines from the skin of the Yangtze finless porpoise

The Yangtze finless porpoise (Neophocaena phocaenoides asiaeorientalis), as the sole freshwater subspecies of N. phocaenoides, is endemic to the Yangtze River and its adjacent lakes. Its population has declined significantly over recent decades. In this study, we established a skin-derived finite fibroblast cell line of the Yangtze finless porpoise, named YFP-SF1, using primary cell culture methods, and an immortalized cell line, T-YFP-SF1, through co-transfection (GFP and SV40 T antigens) techniques. YFP-SF1 proliferated continuously with a minimum population doubling time of 31h and exhibited age-dependent changes in growth rate. T-YFP-SF1 cells exhibited fibroblast morphology and were characterized by a shorter doubling time, higher attachment efficiencies, colony formation at a low seeding density, and growth in low serum concentrations. Anchorage independence and foci formation in the cell monolayer were observed from passage 36. The chromosome number of YFP-SF1 and T-YFP-SF1 remained stable at 2n = 44 in the early passages, and the viability of thawed cells remained above 90% after cryopreservation in liquid nitrogen. Taken together, we have established fibroblast cell lines of Yangtze finless porpoise for the first time, which might assist as an in vitro model for this endangered mammal.

Physiological responses to capture and handling of free-ranging male Yangtze finless porpoises (Neophocaena phocaenoides asiaeorientalis)

We captured free-ranging male Yangtze finless porpoises over three seasons and assayed leukocytes and serum biochemistry to investigate physiological responses to the capture and handlings. Serum thyroid hormones (THs) declined sharply in those porpoises compared with hormone variation in a captive male finless porpoise. Hypernatremia and hypokalemia were also significant in the free-ranging animals suggesting that conservation of serum sodium might be acutely vital for this freshwater subspecies. The animals captured in spring showed more significant neutrophilia and eosinopenia than those captured in autumn suggesting that they may be more affected by capture during the breeding season. Furthermore, physical examination of porpoises when out of the water was apparently stressful, particularly when they were kept out of the water for longer periods. However, an increase in circulating THs may be an adaptive response to accommodate these short-term stresses.

Preliminary hazard assessment of polychlorinated biphenyls, polybrominated diphenyl ethers, and polychlorinated dibenzo‐p‐dioxins and dibenzofurans to yangtze finless porpoise in dongting lake, china

Yangtze finless porpoise (Neophocaena phocaenoides asiaeorientalis), a protected endangered species, is the sole freshwater subspecies of finless porpoise, living only in the middle and lower reaches of the Yangtze River, China, and its appended lakes. Its population has decreased sharply to 1,400 because of human activities, including environmental contamination. In the present study, polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs), and polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) were determined in the blubber, liver, kidney, stomach, small intestine, and brains of five individual Yangtze finless porpoises collected from 1998 to 2004. The results showed PCB concentrations ranged from 0.06 to 1.89 microg/g lipid weight in the organs and consisted mainly of penta-, hexa-, and decachlorinated biphenyls. The PBDE concentrations were between 5.32 and 72.76 ng/g lipid weight. Tetra-, penta-, and hexabrominated diphenyl ethers were the major homologues. The PCDD/F concentrations ranged from 65 to 1,563 pg/g lipid weight, and their predominant homologues were penta- and hexachlorinated dibenzofurans and hepta- and octachlorinated dibenzo-p-dioxins. The hazard quotients (HQs) based on toxic equivalency were determined to be greater than one in all individuals for PCBs, for PCDD/Fs, and for PCBs and PCDD/Fs In addition, HQs would be higher if PBDEs were included. The results suggest that reduction of environmental contamination may contribute greatly to protecting this highly endangered species.

Phylogeography and environmental diversification of a highly adaptable marine amphipod, Gammarus duebeni

Genetic diversity and phylogeographic population structure in the gammarid amphipod, Gammarus duebeni, were investigated across its broad latitudinal distribution in the NE and NW Atlantic by analysis of mitochondrial DNA sequence. Gammarus duebeni has exceptional tolerance of salinity change and inhabits environments ranging from marine to freshwater. The longstanding debate on whether there are distinct marine and freshwater subspecies was assessed by sampling populations from sites characterized by different salinities. Our sequence data demonstrates that there are two major lineages, with little internal geographic structuring. Evidence is provided to suggest a pre-glacial divergence of these two clades, involving segregation between a region historically associated with the freshwater form and the majority of the marine localities on both sides of the Atlantic. A modern contact zone between the marine and freshwater forms is proposed in western Britain.

Aquatic Resource Conservation. The first Yangtze finless porpoise successfully born in captivity (4 pp)

As the sole freshwater subspecies of finless porpoise (Neophocaena phocaenoides), the Yangtze finless porpoise (N. p. asiaeorientalis) lives only in the middle and lower reaches of the Yangtze River and its appended Poyang and Dongting Lakes. As a result of human activity on the river, including over and illegal fishing, pollution, transportation and dam construction, the population of Yangtze finless porpoises has been steadily and rapidly decreasing during the past several decades, which leads the animal to be endangered. For saving this unique animal from extinction, three corresponding measures, in situ conservation, ex situ conservation, and intensifying breeding research in captivity, were proposed and have been implemented since the 1980s. After successfully rearing the animals in captivity for almost nine years, the first Yangtze finless porpoise was successfully born in captivity on July 5, 2005. The calf is male, with a body length of 69 cm. This is the first freshwater cetacean ever born in captivity. The successful birth of this calf confirms that it is possible to breed the Yangtze finless porpoise in captivity. Furthermore, this will greatly benefit the conservation efforts, and also greatly bolster our on-going efforts to study the reproductive biology of these animals. More studies and efforts are expected to establish a sustainable, captive colony of the Yangtze finless porpoise, which will not only greatly expand our knowledge about the reproduction biology of this animal, but also help to redeem the wild population through a careful yearly 'soft releasing' process.

Genetic differentiation between the freshwater subspecies of Gasterosteus aculeatus in Southern California

Samples of Gasterosteus aculeatus microcephalus and the endangered G. aculeatus williamsoni were obtained from downstream and headwaters regions, respectively, of the Santa Clara River system in southern California and electrophoretically examined for the products of 45 loci, six of which were polymorphic. Significant departures from Hardy-Weinberg expectations were evident in genotypic distributions at four of the polymorphic loci indicating a severe restriction of gene flow within the drainage. The genetic distance between these two subspecies was calculated as D = 0.034 ± 0.029, a value comparable to that reported between G. aculeatus microcephalus and G. aculeatus aculeatus from different drainages in northern California.

Evolution of Phenotypic Diversity in Gasterosteus aculeatus Superspecies on the Pacific Coast of North America

Bell, M. A. (Department of Biology, University of California, Los Angeles, Califomnia 90024) 1976. Evolution of phenotypic diversity in Gasterosteus aculeatus superspecies on the Pacific coast of North America. Syst. Zool. 25:211-227.-A conceptual model for the evolution of phenotypic diversity of the threespine stickleback on the Pacific coast of North America is presented. This model takes into account biological characteristics of the threespine sticklebackand climatic changes during Pleistocene times. Biological features considered include: (1) life history patterns in freshwater and anadromous populations; (2) distribution of freshwater and anadromous populations; (3) morphological variation in freshwater populations; (4) genetics of morphological features; (5) the characteristics of highly divergent populations and their distribution in time and space; (6) character displacement. Previous models for the evolution of phenotypic diversity and the interactions of selection and gene flow are considered. Hypotheses concerning protein evolution in Pacific coast sticklebacks are generated from the model. These hypotheses depend on recent evidence that protein evolution does not have a cause and effect relationship with phenotypic divergence and speciation. A causal relationship between the unusual mode of evolution in western North American Gasterosteus and their low haploid DNA content is tentatively suggested. If the model is correct, freshwater subspecies proposed by Miller and Hubbs (1969) for the nominal species Gasterosteus aculeatus are polyphyletic. [Teleostei; Gasterosteidae; speciation.] Gasterosteus aculeatus, the threespine stickleback, appears to be a superspecies composed of semispecies, some of which are polytypic and polymorphic but all of which form a cohesive taxonomic unit. This remarkable diversity led first to an explosive proliferation of nominal species followed by a process of synonymy. Taxonomic questions aside, the evolutionary mechanisms responsible for this phenotypic diversity, restricted within bounds that have not been breached to produce progressive adaptive radiation leading to distinctive, divergent lineages must be complex. It is generally believed that the biological species concept articulated by Mayr (1969) adequately describes the attributes of taxa regarded as species. This concept of species, however, fails to explain the situation characterized by the threespine stickleback. But is the threespine stickleback a rare exception to a generally valid rule? It could be that the threespine stickleback more clearly displays the genetic diversity, which, unperceived in other organisms, has led us to an oversimplified view of the evolutionary unit represented by species. According to the biological species concept, the bond holding together species is gene flow. Among other attributes ascribed to the biological species, Mayr (1969) considered the biological species to be .. . a genetic unit consisting of a large, intercommunicating gene pool,... We find that in Gasterosteus aculeatus, in the absence of gene flow for periods sufficient to allow morphological divergence, morphological divergence seems almost inevitable but usually is held within well defined limits. The vast majority of populations are composed of one or more of Hagen and Gilbertson's (1972) morphs (i.e., low, partial and completely plated), and this is an adequate characterization of them. The criteria for biological species may or may not be attained by divergent populations included within these limits. Populations exceeding these limits may attain the status of biological species, but even among such populations, their phenotypic characteris-

The Variability of the Dimensions of the Merus of the Fifth Pereiopod in the Amphipod Gammarus Duebeni Liljeborg, 1852

The variability of the morphological character on which the distinction between a brackish water and a freshwater subspecies of Gammarus duebeni is based, viz. the ratio merus width/merus length in the fifth walking leg, is studied. Evidence is given, that the merus dimensions only very partially can be correlated with the distribution and the ecology of G. duebeni. No clear clinal variation can be found in the merus dimensions in Europe. The subspecies G. d. celticus is found only in inland waters in Ireland and Brittany; the subspecies G. d. duebeni is found in western and northwestern Europe, and in Canada, in brackish water habitats and in some freshwater localities. Evidence is given, that G. d. duebeni has invaded the latter freshwater localities only recently.

Calcium and other ions in blood and skeleton of Nicaraguan fresh-water shark.

The bull shark, Carcharhinus leucas, employing archaic but effective means of regulating the physical-chemical composition of its body fluids, thrives in tropical fresh-water rivers and lakes. The ionic strength of the serum and the concentrations of total solutes, calcium, urea, and other ions are below the levels found in marine elasmobranchs but higher than the levels in teleosts. The patterns of the calcium deposits of the vertebrae are identical in marine and fresh-water subspecies.

Calcium and Other Ions in Blood and Skeleton of Nicaraguan Fresh-Water Shark

The bull shark, Carcharhinus leucas, employing archaic but effective means of regulating the physical-chemical composition of its body fluids, thrives in tropical fresh-water rivers and lakes. The ionic strength of the serum and the concentrations of total solutes, calcium, urea, and other ions are below the levels found in marine elasmobranchs but higher than the levels in teleosts. The patterns of the calcium deposits of the vertebrae are identical in marine and fresh-water subspecies.

Calcium and Other Ions in Blood and Skeleton of Nicaraguan Fresh-Water Shark

The bull shark, Carcharhinus leucas, employing archaic but effective means of regulating the physical-chemical composition of its body fluids, thrives in tropical fresh-water rivers and lakes. The ionic strength of the serum and the concentrations of total solutes, calcium, urea, and other ions are below the levels found in marine elasmobranchs but higher than the levels in teleosts. The patterns of the calcium deposits of the vertebrae are identical in marine and fresh-water subspecies.