Mikrotxipak arrainetan: atzo, gaur eta bihar

Microarrays have been designed among others, to study transcriptomic differences or differences in the mRNA that is being transcribed, between target organisms, tissues or cells. As result, the transcriptional mode of action of different stress factors are been widely studied in aquatic organisms. Thus, the principal aim of this manuscript, is to summarize fish microarray studies that reveal the mode of action of chemical substances typically found in aquatic environments, as well as, to point out microarray studies derived criticisms.; Mikrotxipak, besteak beste, itu organismo, ehun edo zelulen artean ematen diren transkriptoma mailako ezberdintasunak ikertzeko diseinatu dira. Emaitza gisa, faktore estresatzaile ezberdinen jokatzeko modua edo jarraibidea itsas organismoetan modu zabalean ikertzea lortu da. Horrela, lan honen helburu nagusiak, batetik, teleosteotan ingurune urtarretan ohikoak diren sustantzia kimikoen ekiteko modua aztertzen duten mikrotxipak laburbiltzea da; eta bestetik, mikrotxipen inguruan dauden kritikak azaltzea

Identification of Sex and Female’s Reproductive Stage in Commercial Fish Species through the Quantification of Ribosomal Transcripts in Gonads

The estimation of maturity and sex of fish stocks in European waters is a requirement of the EU Data Collection Framework as part of the policy to improve fisheries management. On the other hand, research on fish biology is increasingly focused in molecular approaches, researchers needing correct identification of fish sex and reproductive stage without necessarily having in house the histological know-how necessary for the task. Taking advantage of the differential gene transcription occurring during fish sex differentiation and gametogenesis, the utility of 5S ribosomal RNA (5S rRNA) and General transcription factor IIIA (gtf3a) in the molecular identification of sex and gametogenic stage was tested in different economically-relevant fish species from the Bay of Biscay. Gonads of 9 fish species (, Atlantic, Atlantic-chub and horse mackerel, blue whiting, bogue, European anchovy, hake and pilchard and megrim), collected from local commercial fishing vessels were histologically sexed and 5S and 18S rRNA concentrations were quantified by capillary electrophoresis to calculate a 5S/18S rRNA index. Degenerate primers permitted cloning and sequencing of gtf3a fragments in 7 of the studied species. 5S rRNA and gtf3a transcript levels, together with 5S/18S rRNA index, distinguished clearly ovaries from testis in all of the studied species. The values were always higher in females than in males. 5S/18S rRNA index values in females were always highest when fish were captured in early phases of ovary development whilst, in later vitellogenic stages, the values decreased significantly. In megrim and European anchovy, where gonads in different oogenesis stages were obtained, the 5S/18S rRNA index identified clearly gametogenic stage. This approach, to the sexing and the quantitative non-subjective identification of the maturity stage of female fish, could have multiple applications in the study of fish stock dynamics, fish reproduction and fecundity and fish biology in general.This work was funded by the University of the Basque Country, Basque Government and the Spanish MINECO through projects: UFI 11/3, S-PE11UN007, S-PE12UN086, S-PE13UN101, IT810-13 and AGL2012-33477. IR is recipient of a grant for predoctoral studies of the Basque Government. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript

Otolith shape and elemental signatures provide insights into the connectivity of euryhaline chelon labrosus inhabiting two close estuaries with different burdens of xenoestrogens in the southern bay of biscay

Intersex gonads have been observed in thicklip grey mullet Chelon labrosus, inhabiting estuaries with high burdens of xenoestrogens in the Southeast Bay of Biscay, but knowledge of population connectivity among estuaries is lacking for this euryhaline fish species. This study investigates the population structure of C. labrosus using otolith shape and elemental signatures of 60 adults (overall length ~ 38 cm) from two estuaries 21 nautic miles apart, one with a high incidence of intersex condition (Gernika), and the other one pristine (Plentzia). Otolith shape analyses were performed using elliptical Fourier descriptors, while elemental signatures of whole sagittae were obtained by inductively coupled plasma mass spectrophotometry. Univariate and multivariate statistics were applied to determine if otolith signatures show patterns of homogeneity between estuaries. The data indicated significant differences in the otolith shape and elemental composition between mullets of Gernika and Plentzia. Elemental differences were mainly driven by Sr, Li (both higher in Plentzia) and Ba (higher in Gernika). The 98% re-classification success rate obtained from stepwise linear discriminant function analysis suggests that Gernika and Plentzia individuals form separated population units. The limited connectivity between these two close estuaries would indicate a different life history of exposure to chemicals, which might explain the high prevalence of intersex condition in Gernika and its absence in Plenztia.info:eu-repo/semantics/publishedVersio

Use of otolith shape and elemental signatures to infer the population structure of the Thicklip Grey Mullet Chelon labrosus in the Southern Bay of Biscay

Xenoestrogenic effects have been reported in thicklip grey mullet, Chelon labrosus, used as pollution sentinel organisms in estuaries in the Southeast Bay of Biscay with intersex gonads described in populations from some contaminated estuaries. Despite evidence of reproductive stress in this catadromous fish species, knowledge of mullet reproductive movements and connectivity between estuaries is lacking. This study investigates the population structure of C. labrosus using otolith shape and elemental signatures of 60 adult individuals collected from two estuaries found in the Southeast Bay of Biscay (Gernika and Plentzia). All samples were collected in June–July 2020. Otolith shape analysis was determined using elliptical Fourier descriptors, while elemental signatures (Sr:Ca, Li:Ca, Mg:Ca, Mn:Ca, Co:Ca, Ni:Ca, Cu:Ca and Ba:Ca) of whole sagittae were determined by inductively coupled plasma mass spectrophotometry. Both natural tags were analyzed with univariate and multivariate statistics to determine whether these signatures are geographically distinct and can be used to assess the degree of separation between individuals. The data showed significant differences in the otolith shape and elemental analyses, with canonical analysis of principal coordinates plots identifying two different groups, each one belonging to each estuary of origin. Differences in whole otolith elemental signatures between locations were driven by Sr:Ca, Li:Ca, and Ba:Ca. Sr:Ca and Li:Ca ratios were higher in Plentzia than in Gernika, while Ba:Ca was higher in Gernika. The high re-classification success rate using both tools obtained from stepwise linear discriminant function analysis supports these findings and suggests that Gernika and Plentzia individuals passed enough time in separated water compartments and should be regarded as two different population units. This could suggest that the intersex condition in mullets from Gernika is due to life-long exposure to xenoestrogens after homing during early larval development in that estuary, without migrations to other estuaries.info:eu-repo/semantics/publishedVersio

High production of transfer RNAs identifies the presence of developing oocytes in ovaries and intersex testes of teleost fish

5S rRNA is highly transcribed in fish oocytes and this transcription levels can be used to identify the presence of oocytes in the intersex testes of fish exposed to xenoestrogens. Similar to 5S rRNA, tRNAs are transcribed by RNA polymerase III (Pol-III) in eukaryotes, so this study focuses in the analysis of the levels of expression of tRNAs in the gonads (ovaries and testes) of eight teleost species as a possible new oocyte molecular marker. Total RNA extracted from gonads of six commercial teleost species in the Biscay Bay, from the pollution sentinel species thicklip grey mullet (Chelon labrosus) known present intersex testes in response to xenoestrogens in Gernika estuary and from the laboratory model species Danio rerio were analysed through capillary electrophoresis. Bioanalyzer electropherograms were used to quantify the concentrations of tRNAs, 5S and 5.8S rRNA. All studied ovaries expressed significantly higher levels of tRNAs and 5S rRNA than testes. A tRNA to 5.8S rRNA index was calculated which differentiates ovaries from testes, and identifies some intersex testes in between testes and ovaries in mullets. The tRNA/5.8S ratio was highest in ovaries in previtellogenic stage, decreasing towards maturity. Thus, strong oocyte expression of tRNAs is an additional proof of high activity levels of Pol-III during early stages of oocyte development in teleost ovaries. Incidentally, we observed that miRNA concentrations were always higher in testes than ovaries. The indexing approach developed in the present study could have multiple applications in teleost reproduction research and in the development of early molecular markers of intersex condition.This work has been supported by project Born2bEgg (Spanish MCIN & EU-FEDER/ERDF (PGC2018-101442-B-100) and by the grants to consolidated research projects of the Basque Government (IT1302-19 and IT1743-22). We are thankful to the Biscay Bay Environmental Biospecimen Bank (BBEBB; PiE-UPV/EHU) for providing mullet tissues

Lack of genetic structure in euryhaline Chelon labrosus from the estuaries under anthropic pressure in the Southern Bay of Biscay to the coastal waters of the Mediterranean Sea

Over the last decade, xenoestrogenic effects have been reported in populations of thicklip grey mullet Chelon labrosus from contaminated estuaries in the Bay of Biscay, resulting in intersex condition. To understand the level of gene flow in individuals of different Basque estuaries microsatellite markers were used to evaluate the population structure and connectivity of C. labrosus from estuaries of the Basque coast. 46 microsatellites were tested and 10 validated for the analysis of 204 individuals collected from 5 selected Basque estuaries and 2 outgroups in the Bay of Cadiz and Thermaic Gulf. The polymorphic microsatellites revealed 74 total alleles, 2–19 alleles per locus. The mean observed heterozygosity (0.49 ± 0.02) was lower than the expected one (0.53 ± 0.01). There was no evidence of genetic differentiation (FST = 0.0098, P = 0.0000) among individuals or sites. Bayesian clustering analysis revealed a single population in all sampled locations. The results of this study indicate widespread genetic homogeneity and panmixia of C. labrosus across the current sampling areas spanning the Atlantic and Mediterranean basins. The hypothesis of panmixia could therefore be well supported so individuals inhabiting estuaries with high prevalence of intersex condition should be considered as members of the same single genetic group as those inhabiting adjacent estuaries without incidence of xenoestrogenicity.This work was funded by the Spanish Ministry of Science. Innovation and Universities and EU-FEDER/ERDF (BORN2bEGG PGC2018-101442-B-I00) and the Basque Government (Grants to consolidated research groups IT1302-19 and IT1743-22). A.N. is a recipient of a pre-doctoral grant PIF17/172 from the University of the Basque Country (UPV/EHU). Authors wish to thank the help of two anonymous reviewers for significantly helping in better focusing the aim of the present work

Antioxidant activities and selenogene transcription in the European sea bass (Dicentrarchus labrax) liver depend, in a non-linear manner, on the Se/Hg molar ratio of the feeds

Feeding 3.9 and 6.7 mg Hg/kg (Se/Hg molar ratios of 0.8 and 0.4, respectively) for 14 days negatively affected Dicentrarchus labrax growth and total DNTB- and thioredoxin-reductase (TrxR) activities and the transcription of four redox genes (txn1, gpx1, txnrd3, and txnrd2) in the liver, but a diet with 0.5 mg Hg/kg (Se/Hg molar ratio 6.6) slightly increased both reductase activities and the transcription of txn1, gpx1, and txnrd2. Feeding 6.7 mg Hg/kg for 53 days downregulated the genes of the thioredoxin system (txn1, txnrd3, and txnrd2) but upregulated gpx1, confirming the previously proposed complementarity among the antioxidant systems. Substitution of 20% of the feed by thawed white fish (hake) slightly counteracted the negative effects of Hg. The effects were not statistically significant and were dependent, in a non-linear manner, on the Se/Hg molar ratio of the feed but not on its Hg concentration. These results stress the need to consider the Se/Hg molar ratio of the feed/food when evaluating the toxicity of Hg.Open Access funding provided thanks to the CRUE-CSIC agreement with Springer Nature. The work was supported by grants from the Spanish MINECO (RTC-2014–2837-2- “SELATUN: Minimización de la problemática del mercurio del atún y valorización del atún como alimento saludable, Programa Retos-Colaboración 2014” and CTM2012-40203-C02-01- “BMW: Biomarcadores estándar de base científica en mejillón para diagnosticar y monitorizar los efectos biológicos de la polución en el Golfo de Bizkaia: implementación de la DEME”), Euskampus Fundazioa-Campus of International Excellence (307615SAA2), and from the Basque Government Elkartek Grant KK-2016/00057- “MAROMEGA: Nuevas alternativas para la producción de omega-3 a partir de recursos marinos.

Antioxidant activities and selenogene transcription in the european sea bass (Dicentrarchus labrax) liver depend, in a non-linear manner, on the Se/Hg molar tatio of the feeds

[EN] Feeding 3.9 and 6.7 mg Hg/kg (Se/Hg molar ratios of 0.8 and 0.4, respectively) for 14 days negatively affected Dicentrarchus labrax growth and total DNTB- and thioredoxin-reductase (TrxR) activities and the transcription of four redox genes (txn1, gpx1, txnrd3, and txnrd2) in the liver, but a diet with 0.5 mg Hg/kg (Se/Hg molar ratio 6.6) slightly increased both reductase activities and the transcription of txn1, gpx1, and txnrd2. Feeding 6.7 mg Hg/kg for 53 days downregulated the genes of the thioredoxin system (txn1, txnrd3, and txnrd2) but upregulated gpx1, confirming the previously proposed complementarity among the antioxidant systems. Substitution of 20% of the feed by thawed white fish (hake) slightly counteracted the negative effects of Hg. The effects were not statistically significant and were dependent, in a non-linear manner, on the Se/Hg molar ratio of the feed but not on its Hg concentration. These results stress the need to consider the Se/Hg molar ratio of the feed/ food when evaluating the toxicity of Hg.The work was supported by grants from the Spanish MINECO (RTC-2014–2837-2- “SELATUN: Minimización de la problemática del mercurio del atún y valorización del atún como alimento saludable, Programa Retos-Colaboración 2014” and CTM2012-40203-C02-01- “BMW: Biomarcadores estándar de base científica en mejillón para diagnosticar y monitorizar los efectos biológicos de la polución en el Golfo de Bizkaia: implementación de la DEME”), Euskampus Fundazioa- Campus of International Excellence (307615SAA2), and from the Basque Government Elkartek Grant KK-2016/00057- “MAROMEGA: Nuevas alternativas para la producción de omega-3 a partir de recursos marinos.” The funding sources had no involvement in the preparation of this manuscript

Mugilid Fish Are Sentinels of Exposure to Endocrine Disrupting Compounds in Coastal and Estuarine Environments

Effects on fish reproduction can result from a variety of toxicity mechanisms first operating at the molecular level. Notably, the presence in the environment of some compounds termed endocrine disrupting chemicals (EDCs) can cause adverse effects on reproduction by interfering with the endocrine system. In some cases, exposure to EDCs leads to the animal feminization and male fish may develop oocytes in testis (intersex condition). Mugilid fish are well suited sentinel organisms to study the effects of reproductive EDCs in the monitoring of estuarine/marine environments. Up-regulation of aromatases and vitellogenins in males and juveniles and the presence of intersex individuals have been described in a wide array of mullet species worldwide. There is a need to develop new molecular markers to identify early feminization responses and intersex condition in fish populations, studying mechanisms that regulate gonad differentiation under exposure to xenoestrogens. Interestingly, an electrophoresis of gonad RNA, shows a strong expression of 5S rRNA in oocytes, indicating the potential of 5S rRNA and its regulating proteins to become useful molecular makers of oocyte presence in testis. Therefore, the use of these oocyte markers to sex and identify intersex mullets could constitute powerful molecular biomarkers to assess xenoestrogenicity in field conditions.This work has been funded through research projects of the Spanish MINECO (SEXOVUM AGL2012-33477), Basque Government (SAIOTEK OVUM-II S-PE12UN086 and Consolidated research groups IT-810-13) and UPV/EHU (UFI 11/37). IRB is recipient of a predoctoral grant of the Basque Government while CB is recipient of a grant of UPV/EHU

Molecular confirmation of pearl formation in arctic mussels (Mytilus edulis) caused by Gymnophallus bursicola (Odhner 1900) metacercariae

Abstract In recent field studies, suspected gymnophallid metacercariae were histologically located in the mantle of mussels from the Norwegian Sea. Mussels from the sites in which that infection was detected also presented abnormally high pearl numbers. It has been previously described that gymnophallid metacercariae could cause pearl formation processes in mussels, as a host reaction to encapsulate these metacercariae. Given the pathological host reaction these parasites elicit, a study was performed to identify gymnophallid metacercariae found in mussels collected from Tromsø at morphological and molecular level and to assess, by the use of molecular tools, the relationship between the parasite and the biological material inside the pearls. As a result, Gymnophallus bursicola metacercariae infecting Norwegian Mytilus edulis were identified according to morphological characters, along with the first 18S rDNA and COI sequences for this trematode species. In addition, parasite DNA from the core of the pearls was extracted and amplified for the first time, confirming the parasitological origin of these pearls. This procedure could allow identifying different parasitic organisms responsible for the generation of pearls in bivalves