Occurrence of a novel acetylated amino acid,Nα-acetylhistidine,in skeletal muscle of freshwater fish and other ectothermic vertebrates

The occurrence of N^α-acetylhistidine (NAH) in skeletal muscle of 91 species of freshwater fish and 9 species of other ectothermic vertebrates was investigated, with consideration of phylogenetic relationships. Of the 91 freshwater fish species examined, 13 species (7 cichlids, 5 anabantids, and 1 catfish) contained considerable amounts (> 1 µmol/g) of NAH in their skeletal muscles. The highest level (10.37 µmol/g) of NAH was found in the tissue of Betta splendens (Siamese fighting fish). Moreover, the NAH contents in the tissues of Trichogaster trichopterus (three spot gourami), Kryptopterus bicirrhis (glass catfish), Oreochromis niloticus (Nile tilapia), Mikrogeophagus ramirezi (ram cichlid) and Parachromis managuensis (Guapote tigre) were 3.17–6.16 µmol/g. The skeletal muscle of amphibians (5 species) and reptiles (4 species) had a low level (< 0.25 µmol/g) of NAH. The present findings clearly demonstrate NAH as the fifth imidazole-related compound, in addition to histidine, carnosine, anserine and ophidine (balenine), recognized as a major non-protein nitrogenous constituent in the skeletal muscle of vertebrate animals.     

Comparative feeding ecology of shortfin mako, blue and thresher sharks in the California Current

This study describes the feeding ecology of three pelagic shark species in the California Current: shortfin mako (Isurus oxyrinchus); blue (Prionace glauca); and thresher (Alopias vulpinus) sharks. Stomach contents of sharks collected from 2002 to 2008 were identified to the lowest taxonomic level and analyzed using univariate and multivariate methods. Of 330 mako sharks sampled (53 to 248?cm fork length [FL]), 238 stomachs contained 42 prey taxa, with jumbo squid (Dosidicus gigas) and Pacific saury (Cololabis saira) representing the most important prey based on the geometric index of importance (GII). In addition, 158 blue sharks were sampled (76 to 248?cm FL) and 114 stomachs contained 38 prey taxa, with jumbo and Gonatus spp. squids representing the most important prey. Lastly, 225 thresher sharks were sampled (108 to 228?cm FL) and 157 stomachs contained 18 prey taxa with northern anchovy (Engraulis mordax) and Pacific sardine (Sardinops sagax) identified as the most important prey. Overall, mako sharks had the most diverse diet based upon Simpson??s diversity index (1/D) (8.43?±?1.16), feeding on many species of teleosts and cephalopods, followed by blue sharks (6.20?±?2.11) which consumed a wide range of prey (primarily cephalopods), while thresher sharks were most specialized (2.62?±?0.34), feeding primarily on coastal pelagic teleosts. Dietary overlap was lowest between blue and thresher sharks (S?rensen similarity index?=?0.321 and Simplified Morisita Horn index?=?0.006), and seasonal variability in diet was greatest for blue sharks (Simplified Morisita Horn index?=?0.260, Analysis of Similarity (ANOSIM) p?<?0.001). In addition, size class, and subregion were significant factors that affected diet of each species differently (ANOSIM p?<?0.001). Despite similarities in life history characteristics and spatial and temporal overlap in habitat, diets of these three common shark species are distinct in the California Current.     

Lampreys Have a Single Gene Cluster for the Fast Skeletal Myosin Heavy Chain Gene Family

Muscle tissues contain the most classic sarcomeric myosin, called myosin II, which consists of 2 heavy chains (MYHs) and 4 light chains. In the case of humans (tetrapod), a total of 6 fast skeletal-type MYH genes (MYHs) are clustered on a single chromosome. In contrast, torafugu (teleost) contains at least 13 fast skeletal MYHs, which are distributed in 5 genomic regions; the MYHs are clustered in 3 of these regions. In the present study, the evolutionary relationship among fast skeletal MYHs is elucidated by comparing the MYHs of teleosts and tetrapods with those of cyclostome lampreys, one of two groups of extant jawless vertebrates (agnathans). We found that lampreys contain at least 3 fast skeletal MYHs, which are clustered in a head-to-tail manner in a single genomic region. Although there was apparent synteny in the corresponding MYH cluster regions between lampreys and tetrapods, phylogenetic analysis indicated that lamprey and tetrapod MYHs have independently duplicated and diversified. Subsequent transgenic approaches showed that the 5′-flanking sequences of Japanese lamprey fast skeletal MYHs function as a regulatory sequence to drive specific reporter gene expression in the fast skeletal muscle of zebrafish embryos. Although zebrafish MYH promoters showed apparent activity to direct reporter gene expression in myogenic cells derived from mice, promoters from Japanese lamprey MYHs had no activity. These results suggest that the muscle-specific regulatory mechanisms are partially conserved between teleosts and tetrapods but not between cyclostomes and tetrapods, despite the conserved synteny.     

Carnosine synthase deficiency is compatible with normal skeletal muscle and olfactory function but causes reduced olfactory sensitivity in aging mice

Carnosine (β-alanyl-l-histidine) and anserine (β-alanyl-3-methyl-l-histidine) are abundant peptides in the nervous system and skeletal muscle of many vertebrates. Many in vitro and in vivo studies demonstrated that exogenously added carnosine can improve muscle contraction, has antioxidant activity, and can quench various reactive aldehydes. Some of these functions likely contribute to the proposed anti-aging activity of carnosine. However, the physiological role of carnosine and related histidine-containing dipeptides (HCDs) is not clear. In this study, we generated a mouse line deficient in carnosine synthase (Carns1). HCDs were undetectable in the primary olfactory system and skeletal muscle of Carns1-deficient mice. Skeletal muscle contraction in these mice, however, was unaltered, and there was no evidence for reduced pH-buffering capacity in the skeletal muscle. Olfactory tests did not reveal any deterioration in 8-month-old mice lacking carnosine. In contrast, aging (18–24-month-old) Carns1-deficient mice exhibited olfactory sensitivity impairments that correlated with an age-dependent reduction in the number of olfactory receptor neurons. Whereas we found no evidence for elevated levels of lipoxidation and glycation end products in the primary olfactory system, protein carbonylation was increased in the olfactory bulb of aged Carns1-deficient mice. Taken together, these results suggest that carnosine in the olfactory system is not essential for information processing in the olfactory signaling pathway but does have a role in the long-term protection of olfactory receptor neurons, possibly through its antioxidant activity.     

Channel catfish (Ictalurus punctatus) protein disulphide isomerase, PDIA6: molecular characterization and expression regulated by bacteria and virus inoculation

Protein disulfide isomerases (PDIs) are thought to aid protein folding and assembly by catalyzing formation and shuffling of cysteine disulfide bonds in the endoplasmic reticulum (ER). Currently, increasing evidence suggests PDIs play an important role in host cell invasion and they are relevant targets for the host immune response. However the roles of specific PDIs in teleosts are little known. Here, we characterized the Protein disulfide isomerase family A, member 6 (PDIA6) from channel catfish, Ictalurus punctatus (named as ccPDIA6). The catfish ccPDIA6 gene was homologous to those of other vertebrate species with 13 exons and 12 introns. The consensus full-length ccPDIA6 cDNA contained an ORF of 1320 bp encoding a putative protein of 439 amino acids. It had a 19 amino acid signal peptide and two active thioredoxin-like domains. Sequence of phylogenic analysis and multiple alignments showed that ccPDIA6 was conserved throughout vertebrate evolution. Southern blot analysis suggested the presence of one copy of the ccPDIA6 gene in the catfish genome. Tissue distribution shows that ccPDIA6 was expressed in all examined tissues at the mRNA level. When using the aquatic zoonotic pathogens such as Edwardsiella tara, Streptococcus iniae, and channel catfish reovirus （CCRV） to challenge channel catfish, ccPDIA6 expression was significant changed in immune-related tissues such as head kidney, intestine, liver and spleen. The results suggested that ccPDIA6 might play an important role in the immunity of channel catfish. This is the first report that the PDI gene may be involved in fish host defense against pathogen infection.     

Predicting occurrence of the fungal symbiote Harpella colonizing black fly larvae in coastal streams of Alabama and Mississippi, USA

The environmental conditions governing symbioses are poorly known in aquatic systems. Stream conditions associated with the distribution of the black fly (Simuliidae) midgut symbiote Harpella were investigated in southern Alabama and Mississippi streams. Stream conditions that were most useful in predicting the distribution of Harpella spp. in the study area were dissolved oxygen and water temperature. Presence of Harpella species in streams was associated with higher dissolved oxygen and decreased water temperature compared to streams where Harpella spp. was absent. Stream conditions associated with the distribution of Harpella spp. in other regions of the world vary according to conditions other than those elucidated here, indicating that geography, host species, and stream conditions play important roles in the spatial distribution of Harpella species.     

Grass carp (Ctenopharyngodon idella) TRAF6 and TAK1: Molecular cloning and expression analysis after Ichthyophthirius multifiliis infection

Ichthyophthirius multifiliis, a pathogenic ciliate parasite, infects almost all freshwater fish species and causes significant economic losses. Tumor necrosis factor receptor-associated factor 6 (TRAF6) and transforming growth factor-β-activated kinase 1 (TAK1) are two important signaling molecules involved in toll-like receptor (TLR) signal transduction. To date, the roles of TRAF6 and TAK1 in host defense against fish parasites are still poorly understood. In the present study, TRAF6 (CiTRAF6) and TAK1 (CiTAK1) were identified from grass carp (Ctenopharyngodon idella). The full-length cDNA sequence of CiTRAF6 (2250 bp) includes an open reading frame (ORF) of 1629 bp, which shows a high similarity to that of Cyprinus carpio TRAF6 and encodes a putative protein of 542 amino acids containing one RING domain, two zinc fingers, one coiled-coil region, and one MATH domain. The full-length CiTAK1 cDNA sequence is 2768 bp and includes an ORF of 1626 bp that encodes a putative protein of 541 amino acids containing a conserved serine/threonine protein kinase catalytic domain and a coiled-coil region. Phylogenetic analysis showed that CiTRAF6 and CiTAK1 were clustered with TRAF6 and TAK1 of other teleosts, respectively. CiTRAF6 and CiTAK1 were both constitutively expressed in all examined tissues but with varied expression levels. The highest expressions of CiTRAF6 and CiTAK1 were in the head kidney and spleen, respectively. The expression profiles of CiTRAF6 and CiTAK1 were detected in grass carp after I. multifiliis infection. Expressions of both genes were significantly up-regulated in the skin, gill, head kidney, and spleen at most time points after infection, indicating that CiTRAF6 and CiTAK1 may play essential roles in grass carp defense against I. multifiliis.     

Quantification of carnosine-related peptides by microchip electrophoresis with chemiluminescence detection

A microchip electrophoresis (MCE) method with chemiluminescence (CL) detection was developed for the determination of carnosine-related peptides, including carnosine, homocarnosine, and anserine, in biological samples. A simple integrated MCE-CL system was built to perform the assays. The highly sensitive CL detection was achieved by means of the CL reaction between hydrogen peroxide and N-(4-aminobutyl)-N-ethylisoluminol-tagged peptides in the presence of adenine as a CL enhancer and Co²⁺ as a catalyst. Experimental conditions for analyte labeling, MCE separation, and CL detection were studied. MCE separation of the above-mentioned three peptides took less than 120 s. Detection limits (signal/noise ratio [S/N] = 3) of 3.0 × 10⁻⁸, 2.8 × 10⁻⁸, and 3.4 × 10⁻⁸ M were obtained for carnosine, anserine, and homocarnosine, respectively. The current MCE-CL method was applied for the determination of carnosine, anserine, and homocarnosine in human cerebrospinal fluid (CSF) and canine plasma. Homocarnosine was detected at the micromolar (μM) level in the CSF samples analyzed, whereas the levels of carnosine and anserine in these samples were below the detection limit of the assay. Interestingly, both carnosine and anserine were detected in the canine plasma samples, whereas homocarnosine was not.     

Immunolocalization of aquaporins 1, 3, and 5 in the nasal respiratory mucosa of a panting species,the sheep (Ovis aries)

The nasal respiratory mucosa is the primary site for evaporative water loss in panting species, necessitating the movement of water across the nasal epithelium. Aquaporins (AQP) are protein channels that facilitate water movement in various fluid transporting tissues of non-panting species. Whether the requirement for enhanced capacity for transepithelial water movement in the nasal respiratory mucosa of panting species has led to differences in AQP localization is unknown. Using immunohistochemistry, we report the localization of AQP1, 3, and 5 in the nasal respiratory mucosa of sheep being exposed to ambient temperatures of ~21 °C or ~38 °C for 4.5 h before death (n=3/treatment). Exposure to either treatment resulted in panting. While exposure to ~38 °C resulted in a higher respiratory frequency (mean difference: 82 breaths min⁻¹; P<0.001) than exposure to ~21 °C, there was no difference in the localization of AQPs. Connective tissue and vascular endothelial cells expressed AQP1. Glandular acini expressed AQP1 and apically localized AQP5, which was also present in glandular duct cells. Ciliated columnar epithelial cells expressed AQP5 apically and AQP3 basolaterally. Basal cells expressed AQP3. The distribution and co-localization of AQPs in the ovine nasal respiratory mucosa is different to that reported in non-panting species and may reflect the physiological demands associated with enhanced respiratory evaporation. We propose that AQP1, 3, and 5 may constitute a transepithelial water pathway via glandular secretions and across the surface epithelium, which provides a possible means for rapid and controllable water movement in the nasal respiratory mucosa of a panting species.     

Nature,origin and possible roles of lipid deposits in Maurolicus muelleri (Gmelin) and Benthosema glaciale (Reinhart) from Ullsfjorden,northern Norway

Summary The lipid-rich pelagic teleost Maurolicus muelleri has large lipid depots located subcutaneously, intramuscularly and around the digestive tract. The lipid is contained within conventional adipocytes and is composed largely of triacylglycerols rich in 20:1 (n-9) and lipid-rich mesopelagic teleost Benthosema glaciale, except that the lipid is predominantly wax esters whose fatty alcohols and fatty acids are both rich in 20:1 (n-9) and 22:1 (n-11) moieties. An origin for the lipids of both species in the wax esters of calanoid copepods is indicated. The anatomical distribution of the lipids in these teleosts and their intracellular location point to their being fundamentally an energy store. The twin roles of neutral lipids in providing metabolic energy and buoyancy are discussed.     

Population genetic structure of Sagittaria natans (Alismataceae), an endangered species in China,revealed by nuclear SSR loci analyses

Sagittaria natans (Alismataceae) is an endangered aquatic herb in China. The nuclear microsatellite (SSR) loci variation of the 11 extant populations of S. natans was investigated in the present study. A relative high level of genetic diversity was found in this species. The Wilcoxon’s signed rank tests did not indicate a recent bottleneck in any of the populations. The Analysis of molecular variance (AMOVA) showed a low level of population differentiation (FST = 0.284) among the populations of S. natans. This result was supported by STRUCTURE analysis (K = 1), the NJ analyses and Mantel test (r = ?0.0221, p > 0.5). The genetic structure could be contributed to an earlier period of more pronounced gene flow when the species had a more continuous distribution. The results of this study could be used as the basis for conservation guidelines for the management of this endangered species in China.     

How diverse are planktonic cryptomonads in Brazil? Advantages and difficulties of a taxonomic-biogeographical approach

We compare ‘temperate data’ with information contained in 49 surveys of cryptomonads from Brazilian tropical regions. These surveys contain a total of 351 records relative to a total of 38 taxa at the species level or below. All taxa were identified using traditional morphological characters visible with the light microscope. Two taxa appear to be new species, and amongst the known species Cryptomonas erosa Ehrenberg is the most frequently recorded one. To date two species of Pseudocryptomonas Bicudo & Tell (all containing multiple photosynthetic endosymbionts/chloroplasts) and the species Cryptomonas brasiliensis Castro, C. Bicudo & D. Bicudo have not been found outside Brazil. With respect to their geographical distribution in Brazilian tropical regions, cryptomonads are better represented in reservoirs and coastal lagoons located in the south-eastern part of the country. Some species appear to be restricted to brackish coastal ecosystems (oligo- to mesohaline and meso- to eutrophic waters). There are only four records of marine cryptomonads. A comparative study of three different kinds of shallow aquatic environments in Brazil – north-eastern reservoirs, a flood-plain Amazonian lake, and brackish lagoons on the south-eastern coast – shows that cryptomonads are to be found constantly, and their species number density, and biomass are generally low (usually <10%) in relation to the other phytoplankton. The case of Plagioselmis Butcher emend. Novarino, Lucas & Morrall, amongst others, exemplifies the great difficulties encountered while attempting to interpret biogeographical information based on light microscopy within the context of a modern systematic and taxonomic framework based on electron microscopy.     

Metabolite Proofreading in Carnosine and Homocarnosine Synthesis: MOLECULAR IDENTIFICATION OF PM20D2 AS β-ALANYL-LYSINE DIPEPTIDASE*

Carnosine synthase is the ATP-dependent ligase responsible for carnosine (β-alanyl-histidine) and homocarnosine (γ-aminobutyryl-histidine) synthesis in skeletal muscle and brain, respectively. This enzyme uses, also at substantial rates, lysine, ornithine, and arginine instead of histidine, yet the resulting dipeptides are virtually absent from muscle or brain, suggesting that they are removed by a “metabolite repair” enzyme. Using a radiolabeled substrate, we found that rat skeletal muscle, heart, and brain contained a cytosolic β-alanyl-lysine dipeptidase activity. This enzyme, which has the characteristics of a metalloenzyme, was purified ≈200-fold from rat skeletal muscle. Mass spectrometry analysis of the fractions obtained at different purification stages indicated parallel enrichment of PM20D2, a peptidase of unknown function belonging to the metallopeptidase 20 family. Western blotting showed coelution of PM20D2 with β-alanyl-lysine dipeptidase activity. Recombinant mouse PM20D2 hydrolyzed β-alanyl-lysine, β-alanyl-ornithine, γ-aminobutyryl-lysine, and γ-aminobutyryl-ornithine as its best substrates. It also acted at lower rates on β-alanyl-arginine and γ-aminobutyryl-arginine but virtually not on carnosine or homocarnosine. Although acting preferentially on basic dipeptides derived from β-alanine or γ-aminobutyrate, PM20D2 also acted at lower rates on some “classic dipeptides” like α-alanyl-lysine and α-lysyl-lysine. The same activity profile was observed with human PM20D2, yet this enzyme was ∼100–200-fold less active on all substrates tested than the mouse enzyme. Cotransfection in HEK293T cells of mouse or human PM20D2 together with carnosine synthase prevented the accumulation of abnormal dipeptides (β-alanyl-lysine, β-alanyl-ornithine, γ-aminobutyryl-lysine), thus favoring the synthesis of carnosine and homocarnosine and confirming the metabolite repair role of PM20D2.     

Microsporidian Species Known To Infect Humans Are Present in Aquatic Birds: Implications for Transmission via Water?

Human microsporidiosis, a serious disease of immunocompetent and immunosuppressed people, can be due to zoonotic and environmental transmission of microsporidian spores. A survey utilizing conventional and molecular techniques for examining feces from 570 free-ranging, captive, and livestock birds demonstrated that 21 animals shed microsporidian spores of species known to infect humans, including Encephalitozoon hellem (20 birds; 3.5%) and Encephalitozoon intestinalis (1 bird; 0.2%). Of 11 avian species that shed E. hellem and E. intestinalis, 8 were aquatic birds (i.e., common waterfowl). The prevalence of microsporidian infections in waterfowl (8.6%) was significantly higher than the prevalence of microsporidian infections in other birds (1.1%) (P < 0.03); waterfowl fecal droppings contained significantly more spores (mean, 3.6 × 10⁵ spores/g) than nonaquatic bird droppings contained (mean, 4.4 × 10⁴ spores/g) (P < 0.003); and the presence of microsporidian spores of species known to infect humans in fecal samples was statistically associated with the aquatic status of the avian host (P < 0.001). We demonstrated that a single visit of a waterfowl flock can introduce into the surface water approximately 9.1 × 10⁸ microsporidian spores of species known to infect humans. Our findings demonstrate that waterborne microsporidian spores of species that infect people can originate from common waterfowl, which usually occur in large numbers and have unlimited access to surface waters, including waters used for production of drinking water.     

Temperature and salinity effects on growth, survival, reproduction, and potential distribution of two non-indigenous botryllid ascidians in British Columbia

Two non-indigenous botryllid ascidian species - Botryllus schlosseri (golden star tunicate) and Botrylloides violaceus (violet tunicate) - have become established in British Columbia (BC), Canada. One species, B. schlosseri, is native to Europe while the other, B. violaceus, is native to Asia. Environmental tolerances of both species are poorly understood. We examined the effects of temperature and salinity on growth, survival, and reproduction of these species in the laboratory in order to characterize their environmental tolerances and preferences. Laboratory-raised juvenile colonies were studied using a two-factorial experimental design with five levels of temperature (5, 10, 15, 20, 25 °C) and five levels of salinity (14, 20, 26, 32, 38‰). Both B. schlosseri and B. violaceus possessed broad temperature and salinity tolerances, but B. schlosseri was slightly more euryhalinal than B. violaceus. Generally, B. schlosseri survived environmental conditions of 10-25 °C and 14-38‰, exhibited positive growth at 10-25 °C and 20-38‰, and attained its largest colony sizes at 15-20 °C and 20-38‰. Botrylloides violaceus tolerated environmental conditions between 5-25 °C and 20-38‰, demonstrated positive growth at 15-25 °C and 26-38‰, and attained its largest colony sizes at 20-25 °C and 26-38‰. Results from the laboratory experiment were then used in a modeling exercise to determine the coastal areas of BC that these organisms might be likely to exist in or invade, based on near-surface temperatures and salinities. The model predicted that no areas were totally unsuitable for survival and growth of either species (based solely on temperature and salinity tolerances), with the most suitable locations being along the west coast of Vancouver Island, a region with significant shellfish aquaculture activity.     

Feeding habits of an endemic fish,Oxygymnocypris stewartii,in the Yarlung Zangbo River in Tibet,China

Feeding habits of Oxygymnocypris stewartii were investigated based on monthly sampling in the Yarlung Zangbo River from August 2008 to August 2009. The gut contents of 194 individuals were analysed and quantified with numerical and gravimetric methods. This species can be considered a generalized and opportunistic predator feeding both on teleosts and aquatic insects. A multivariate analysis revealed noticeable variation in O. stewartii diet composition with fish size and season. The smaller specimens fed primarily on Cobitidae and Hydropsychidae. As they grew, Cyprinidae and Chironomidae larvae became important prey. The preferred food items were teleosts in summer and autumn. For winter and spring, the predominant prey switched to chironomidae larvae. Diet composition did not significantly vary between the sexes. Finally, a significant and positive correlation between predator and prey length was found. These findings provide the fundamental information better understanding the role of this important endemic species in the Yarlung Zangbo River food web.     

Pancreatonema torriensis gen. nov., sp. nov. (Nematoda: Rhabdochonidae) from the pancreatic duct of Raja naevus

A new genus and species of nematode, Pancreatonema torriensis, from the pancreatic duct of Raja naevus caught off Aberdeen and north-west Scotland is described and aspects of its biology are discussed. According to the classification of Skrjabin, Sobolev & Ivashkin (1967) [Spirurata of animals and man and the diseases caused by them (4). Essentials of Nematodology, Vol. 16 (I.P.S.T. Translation No. 5904, Jerusalem, 1971)]it belongs to the family Rhabdochonidae, sub-family Prosungulonematinae, and is closely related to the genus Johnstonmawsonia. The taxonomy of the Prosungulonematinae is confused but it is suggested that Johnstonmawsonia should at present be restricted to those species of the genus described from marine teleosts, and that the species from freshwater teleosts should be included in the genus Prosungulonema.     

Specificity and distribution of mammalian carnosinase.

Hog kidney carnosinase (EC 3.4.13.3) was found to have a narrow specificity; it hydrolyzed carnosine, anserine and glycyl-L-histidine, but did not split L-alanyl-L-histidine or homocarnosine. The isoelectric point of this enzyme was 5.8 and its molecular weight was about 84 000. Carnosinase was found to be widely distributed in various tissues of the rat. Uterus, kidney, liver and lung contained high levels of carnosinase, whereas moderate concentrations were found in spleen, heart and brain, with low levels in small intestine, skeletal muscle and stomach, and none in blood.     

Pathogenic Chytrid Fungus Batrachochytrium dendrobatidis,but Not B. salamandrivorans,Detected on Eastern Hellbenders

Recent worldwide declines and extinctions of amphibian populations have been attributed to chytridiomycosis, a disease caused by the pathogenic fungus Batrachochytrium dendrobatidis (Bd). Until recently, Bd was thought to be the only Batrachochytrium species that infects amphibians; however a newly described species, Batrachochytrium salamandrivorans (Bs), is linked to die-offs in European fire salamanders (Salamandra salamandra). Little is known about the distribution, host range, or origin of Bs. In this study, we surveyed populations of an aquatic salamander that is declining in the United States, the eastern hellbender (Cryptobranchus alleganiensis alleganiensis), for the presence of Bs and Bd. Skin swabs were collected from a total of 91 individuals in New York, Pennsylvania, Ohio, and Virginia, and tested for both pathogens using duplex qPCR. Bs was not detected in any samples, suggesting it was not present in these hellbender populations (0% prevalence, 95% confidence intervals of 0.0–0.04). Bd was found on 22 hellbenders (24% prevalence, 95% confidence intervals of 0.16 ≤ 0.24 ≤ 0.34), representing all four states. All positive samples had low loads of Bd zoospores (12.7 ± 4.9 S.E.M. genome equivalents) compared to other Bd susceptible species. More research is needed to determine the impact of Batrachochytrium infection on hellbender fitness and population viability. In particular, understanding how hellbenders limit Bd infection intensity in an aquatic environment may yield important insights for amphibian conservation. This study is among the first to evaluate the distribution of Bs in the United States, and is consistent with another, which failed to detect Bs in the U.S. Knowledge about the distribution, host-range, and origin of Bs may help control the spread of this pathogen, especially to regions of high salamander diversity, such as the eastern United States.     

Global diversity of dipteran families (Insecta Diptera) in freshwater (excluding Simulidae, Culicidae, Chironomidae, Tipulidae and Tabanidae)

Today’s knowledge of worldwide species diversity of 19 families of aquatic Diptera in Continental Waters is presented. Nevertheless, we have to face for certain in most groups a restricted knowledge about distribution, ecology and systematic, particularly in the tropical environments. At the same time we realize a dramatically decline or even lack of specialists being able, having the time or the opportunity to extend or even secure the present information. The respective families with approximate numbers of aquatic species are: Blephariceridae (308), Deuterophlebiidae (14), Nyphomyiidae (7), Psychodidae (∼2.000), Scatopsidae (∼5), Tanyderidae (41), Ptychopteridae (69), Dixidae (173), Corethrellidae (97), Chaoboridae (∼50), Thaumaleidae (∼170), Ceratopogonidae (∼6.000), Stratiomyidae (∼43), Empididae (∼660), Lonchopteridae (2), Syrphidae (∼1.080), Sciomyzidae (∼190), Ephydridae (∼1.500), Muscidae (∼870). Numbers of aquatic species will surely increase with increased ecological and taxonomical efforts. Guest editors: E. V. Balian, C. Lévêque, H. Segers & K. Martens Freshwater Animal Diversity Assessment