Fish pathogens near the Arctic Circle: molecular,morphological and ecological evidence for unexpected diversity of Diplostomum (Digenea: diplostomidae) in Iceland

Host–parasite systems at high latitudes are promising model systems for detecting and predicting the impact of accelerated environmental change. A major challenge is the lack of baselines for the diversity and distribution of parasites in Arctic wildlife, especially in the freshwater environment. Here we present the first known estimates of the species diversity and host associations of Diplostomum spp. in sub-Arctic freshwater ecosystems of the Palaearctic. Our analyses integrating different analytical approaches, phylogenies based on mitochondrial and nuclear DNA, estimates of genetic divergence, character-based barcoding, morphological examination, precise detection of microhabitat specialisation and host use, led to the discovery of one described and five putative new species that complete their life-cycles within a fairly narrow geographic area in Iceland. This increases the species richness of Diplostomum in Iceland by 200% and raises the number of molecularly characterised species from the Palaearctic to 17 species. Our results suggest that the diversity of Diplostomum spp. is underestimated globally in the high latitude ecosystems and call for a cautionary approach to pathogen identification in developing the much needed baselines of pathogen diversity that may help detect effects of climate change in the freshwater environment of the sub-Arctic.     

Solution and high‐pressure NMR studies of the structure,dynamics, and stability of the cross‐reactive allergenic cod parvalbumin Gad m 1

Beta‐parvalbumins from different fish species have been identified as the main elicitors of IgE‐mediated reactions in fish‐allergic individuals. Here, we report for the first time the NMR determination of the structure and dynamics of the major Atlantic cod (Gadus morhua) allergen Gad m 1 and compare them with other known parvalbumins. Although the Gad m 1 structure and accessibility of putative IgE epitopes are similar to parvalbumins in mackerel and carp, the charge distribution at the putative epitopes is different. The determination of the Gad m 1 structure contributes to a better understanding of cross‐reactivity among fish parvalbumins. In addition, the high‐pressure NMR and temperature variation experiments revealed the important contribution of the AB motif and other regions to the protein folding. This structural information could assist the future identification of hot spots for targeted mutations to develop hypoallergenic Ca²⁺‐free forms for potential use in immunotherapy. Proteins 2014; 82:3032–3042. © 2014 Wiley Periodicals, Inc.     

GW501516-activated PPARβ/δ promotes liver fibrosis via p38-JNK MAPK-induced hepatic stellate cell proliferation

Background

After liver injury, the repair process comprises activation and proliferation of hepatic stellate cells (HSCs), which produce extracellular matrix (ECM) proteins. Peroxisome proliferator-activated receptor beta/delta (PPARβ/δ) is highly expressed in these cells, but its function in liver repair remains incompletely understood. This study investigated whether activation of PPARβ/δ with the ligand GW501516 influenced the fibrotic response to injury from chronic carbon tetrachloride (CCl₄) treatment in mice. Wild type and PPARβ/δ-null mice were treated with CCl₄ alone or CCl₄ co-administered with GW501516. To unveil mechanisms underlying the PPARβ/δ-dependent effects, we analyzed the proliferative response of human LX-2 HSCs to GW501516 in the presence or absence of PPARβ/δ.

Results

We found that GW501516 treatment enhanced the fibrotic response. Compared to the other experimental groups, CCl₄/GW501516-treated wild type mice exhibited increased expression of various profibrotic and pro-inflammatory genes, such as those involved in extracellular matrix deposition and macrophage recruitment. Importantly, compared to healthy liver, hepatic fibrotic tissues from alcoholic patients showed increased expression of several PPAR target genes, including phosphoinositide-dependent kinase-1, transforming growth factor beta-1, and monocyte chemoattractant protein-1. GW501516 stimulated HSC proliferation that caused enhanced fibrotic and inflammatory responses, by increasing the phosphorylation of p38 and c-Jun N-terminal kinases through the phosphoinositide-3 kinase/protein kinase-C alpha/beta mixed lineage kinase-3 pathway.

Conclusions

This study clarified the mechanism underlying GW501516-dependent promotion of hepatic repair by stimulating proliferation of HSCs via the p38 and JNK MAPK pathways.     

Effects of fishing on parasitism in a sparid fish: contrasts between two areas of the Western Mediterranean

This study addressed the impacts of fishing on the rates of parasitism using the sparid Boops boops as a model fish species. Using a large suite of parasite species in B. boops, with different life histories, transmission pathways and host specificity, we compared parasite diversity, prevalence, abundance and community structure at two Mediterranean localities in the Balearic Sea, Santa Pola Bay and the Gulf of Oran, that are characterised by a contrasting pattern of fishing of B. boops. A total of 360 fish were examined comprising nine distinct samples collected during the warm and the cold weather months. A total of 29 parasite species were identified, with eight species in common for the two localities. Parasite component communities at Santa Pola Bay were more species rich and abundant than those at the Gulf of Oran and exhibited a different community structure. Of the eight common taxa used in the quantitative comparisons, five exhibited significant difference for prevalence between the two localities, four having substantially higher prevalence at Santa Pola and only one being more prevalent at the Gulf of Oran. Two specialist trematodes and the sparid generalist monogenean exhibited consistently higher prevalence and abundance at Santa Pola Bay than at the Gulf of Oran; the two specialists were also identified as key species for assigning individual fish to their locality of origin. The consistent differences in the richness, abundance and structure of parasite communities in B. boops from Santa Pola Bay and the Gulf of Oran may reflect the contrasting patterns of exploitation of the populations of this fish host at the two localities.     

Random forests, a novel approach for discrimination of fish populations using parasites as biological tags

Due to the complexity of host-parasite relationships, discrimination between fish populations using parasites as biological tags is difficult. This study introduces, to our knowledge for the first time, random forests (RF) as a new modelling technique in the application of parasite community data as biological markers for population assignment of fish. This novel approach is applied to a dataset with a complex structure comprising 763 parasite infracommunities in population samples of Atlantic cod, Gadus morhua, from the spawning/feeding areas in five regions in the North East Atlantic (Baltic, Celtic, Irish and North seas and Icelandic waters). The learning behaviour of RF is evaluated in comparison with two other algorithms applied to class assignment problems, the linear discriminant function analysis (LDA) and artificial neural networks (ANN). The three algorithms are used to develop predictive models applying three cross-validation procedures in a series of experiments (252 models in total). The comparative approach to RF, LDA and ANN algorithms applied to the same datasets demonstrates the competitive potential of RF for developing predictive models since RF exhibited better accuracy of prediction and outperformed LDA and ANN in the assignment of fish to their regions of sampling using parasite community data. The comparative analyses and the validation experiment with a 'blind' sample confirmed that RF models performed more effectively with a large and diverse training set and a large number of variables. The discrimination results obtained for a migratory fish species with largely overlapping parasite communities reflects the high potential of RF for developing predictive models using data that are both complex and noisy, and indicates that it is a promising tool for parasite tag studies. Our results suggest that parasite community data can be used successfully to discriminate individual cod from the five different regions of the North East Atlantic studied using RF.     

Saturnius minutus n. sp. and S. dimitrovi n. sp. (Digenea: Hemiuridae) from Mugil cephalus L. (Teleostei: Mugilidae), with a multivariate morphological analysis of the Mediterranean species of Saturnius Manter, 1969

Three species of the bunocotyline genus Saturnius Manter, 1969 are described from the stomach lining of mugilid fishes of the Mediterranean and Black Seas. Two of the species are new: S. minutus n. sp. occurs in Mugil cephalus off the Mediterranean coast of Spain; and S. dimitrovi n. sp., a parasite of M. cephalus off the Bulgarian Black Sea coast and the Spanish Mediterranean coast, was originally described as S. papernai by Dimitrov et al. (1998). In addition, S. papernai Overstreet, 1977 is redescribed from M. cephalus off the Spanish Mediterranean coast and from Liza aurata and L. saliens off the Bulgarian Black Sea coast. The three species are distinguished morphometrically using univariate and multivariate analyses. These results were verified using Linear Discriminant Analysis which correctly allocated all specimens to their species designations based on morphology (i.e. 100% successful classification rate) and assigned almost all specimens to the correct population (locality). The following variables were selected for optimal separation between samples: the length of the forebody, ventral sucker and posterior testis, the length and width of the posteriormost pseudosegment, and the width of the muscular flange at ventral sucker level.     

A revision of Petasiger Dietz, 1909 (Digenea: Echinostomatidae) and a key to its species

Petasiger Dietz, 1909 is revised, and a key to and lists of the records and hosts of the 18 recognised species are presented. The recognised species are: P. exaeretus Dietz, 1909, P. australis Johnston & Angel, 1941, P. combesi Zamparo, Overstreet & Brooks, 2005, P. floridus Premvati, 1968, P. grandivesicularis Ishii, 1935, P. islandicus Kostadinova & Skirnisson, 2007, P. johnstoni n. sp., P. megacanthus (Kotlán, 1922), P. minutissimus Gogate, 1934, P. neocomense Fuhrmann, 1927, P. nitidus Linton, 1928, P. novemdecim Lutz, 1928, P. oschmarini Kostadinova & Gibson, 1998, P. phalacrocoracis (Yamaguti, 1939), P. pseudoneocomense Bravo-Hollis, 1969, P. pungens (Linstow, 1894), P. soochowensis Ku, Chiu, Li & Zhu, 1977 and P. variospinosus (Odhner, 1910). Seven of these species are redescribed on the basis of type and newly collected material. P. laricola Ku, Chiu, Li & Zhu, 1977, Echinochasmus fotedari Chishti & Mir, 1989 and P. neocomense of Cankovi?, Kazi? & Milka (1983) are considered synonyms of P. pungens; P. longicirratus Ku, 1938, P. skrjabini Bashkirova, 1941 and P. nitidus of Chen et al. (1985) synonyms of P. neocomense; P. caribbensis Nassi, 1980 a synonym of P. novemdecim; and P. tientsinensis Ku, Chiu, Li & Zhu, 1977 a synonym of P. exaeretus. P. johnstoni n. sp. is described from Tachybaptus ruficollis novaehollandiae in Australia. It is readily distinguished from the most closely-related P. grandivesicularis Ishii, 1935 and P. australis Johnston & Angel, 1941 by the smaller size of the body and angle collar spines and differences in most other metrical characters. Species attributed to Petasiger which are here considered species inquirendae are: P. antigonus Nigam, 1944 sp. inq., P. chandleri Abdel-Malek, 1952 sp. inq., P. jubilarum Elperina in Skrjabin, Petrov & Bashkirova, 1947 sp. inq., P. nicolli Pande, 1939 sp. inq. and P. spasskyi Oshmarin in Skrjabin, Petrov & Bashkirova, 1947 sp. inq.; and Echinoparyphium inopinatum (Baer, 1959) n. comb. is a species previously attributed to Petasiger.     

Experimental data on the life-cycle of Petasiger grandivesicularis Ishii, 1935 (Trematoda: Echinostomatidae)

Larval stages of an echinostome were found in Planorbis planorbis in a brackish water lake on the Black Sea coast of Bulgaria. The cercaria is a large-tailed form with 19 collar spines. The life-cycle was completed in the laboratory using aquarium-reared fishes (Lebistes reticulatus, Puntius tetrazona tetrazona, P. pentazona pentazona, P. nigrofasciatus, Carassius auratus auratus and Xiphophorus helleri) as second intermediate hosts and canaries as definitive hosts. The redia, cercaria, metacercaria and experimentally reared adults are described. The species is determined as Petasiger grandivesicularis Ishii, 1935, and its cercaria is compared in detail with those of related forms. A key to the known large-tailed echinostome cercariae from the Palaearctic Region is presented.     

Mining parasite data using genetic programming

Genetic programming is a technique that can be used to tackle the hugely demanding data-processing problems encountered in the natural sciences. Application of genetic programming to a problem using parasites as biological tags demonstrates its potential for developing explanatory models using data that are both complex and noisy.