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Does hydrogen-rich water mitigate MP toxicity in rainbow trout (Oncorhyncus mykiss)? Monitoring with hematology, DNA damage, and apoptosis via ROS/GSH/MDA pathway

Atamanalp Muhammed, Kırıcı Muammer, Köktürk Mine, Kirici Mahinur, Alwazeer Duried, Kocaman Esat Mahmut, Ucar Erzu, Parlak Veysel, Özcan Sinan, Alak Gonca

Oceanological and Hydrobiological Studies

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2023

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Vol. 52, No. 2

206--220

EN

Although the number of studies documenting the presence of Microplastics (MP) in fish is increasing, research studies focused on its detoxification are very limited. In this study, rainbow trout ( Oncorhyncus mykiss) were randomly divided into two groups after being fed with MPs (15% polypropylene [PP] +15% polyethylene [PE]) for 2 months. MP excretion without any application (PP+PE) in group I fish, and excretion of MPs with hydrogen-rich water (HRW) application (PP+PE+HRW) in group 2 were investigated under semi-static conditions for 21 days. This effectwas also compared by using positive and negative control groups (Control[no treatment, free PP, PE or/and HRW] and only HRW group). In thisdirection, the following were determined: PP+PE chronic toxicity in aquatic organisms, the toxicity mechanism and the effect of HRW as a possibletreatment method in blood tissue; with hematological indices ([RBC count [RBC], leukocyte count [WBC], hemoglobin value [Hb], hematocrit ratio [Hct], platelet count [PLT], hemoglobin count per erythrocyte [MCHC],mean hemoglobin amount per erythrocyte [MCH] and mean erythrocyte volume [MCV]) in other tissues (liver, gill and brain tissue) oxidative stress response (catalase [CAT]), glutathione peroxidase (GPx), glutathione reductase (GR), superoxide dismutase (SOD), glutathione (GSH), reactive oxygen products (ROS), malondialdehyde ([MDA] levels), DNA damage (8-OHdG: 8-Hydroxy-2-Deoxyguanosine), and the apoptosis (caspase 3) levels were investigated. In addition, acetylcholinesterase enzyme (AChE) activity, which is important in neurotoxicity pathways in the brain, wasdetermined. The presence of plastics (PP/PE) in target tissues (muscle, liver,gill and gastrointestinal tract) was also obtained.The results showed that PP+PE caused toxicity in all three tissues. MPs showed an inhibiting effect on antioxidant enzyme activities and an inductive effect on MDA, ROS, 8-OHdG, and caspase 3 levels. HRW showed a mitigating effect on MP-mediated toxicity in O. mykiss brain, blood, gill, and liver by controlling the ROS/ GSH/MDA pathway. HRW can be suggested as a cost-effective and eco-friendly curative for the protection of fish from the oxidative damages produced by the ingestion of microplastics.

2

Methane Production from Farmed Rainbow Trout Byproducts by Anaerobic Digestion in Morocco

Essalhi Fatima, Bengueddour Rachid

Ecological Engineering & Environmental Technology

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2023

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Vol. 24, iss. 2

45--54

EN

Fish are a staple in human nutrition because of their great nutritional and dietetic value. The processing stages of these fish before being sold generates an important quantity of byproducts (head, viscera, skeleton, fins, scales, tail, and skin). This study aims to determine the methane production, methanogenic potential and the percentage of volatile solids removed by the anaerobic digestion process. The experiment was carried out at mesophilic conditions in a Continuous Stirred-Tank Reactor (CSTR) with a discontinuous feeding mode (batch). The results obtained showed that the byproducts of farmed rainbow trout have a maximum methane production 1176 Nml whose methanogenic potential is 206.68 Nml/gVS with a biodegradability equal to 57.95%. For the kinetic modeling, four models were applied for an adequate fit of the experimental results (first order, MGompertz, transfererence function, and logistic function). The production of methane closest to the experimental results is that estimated by the logistic function with a methanogenic potential of 212.21 Nml/gVS whose correlation coefficient R2= 0.9870 and a very low percentage of error (1.18%), also the MGompertz model presented results adapted to those of the experiment with a methanogenic potential equal to 223.61 Nml/gVS (R2= 0.9889 and %error = 2.95); This confirms that these two kinetic models are the most suitable for this type of substrate.

3

Wykorzystanie metody QIM do oceny świeżości pstrąga tęczowego (Oncorhynchus Mykiss Walb.) przechowywanego w warunkach chłodniczych

Litwińczuk A., Skałecki P., Florek M., Staszowska A., Kaliniak A., Zaborska A.

Zeszyty Naukowe Akademii Morskiej w Gdyni

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2014

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nr 86

252--257

PL

W pracy oceniono za pomocą systemu QIM wyróżniki sensoryczne 30 pstrągów tęczowych w czasie przechowywania chłodniczego przez 48 godz. Ponadto dokonano instrumentalnych pomiarów właściwości fizykochemicznych tkanki mięśniowej: pH, przewodności elektrycznej właściwej i barwy skóry (wg CIE L*a*b*). Indeks (QI) po uśmierceniu ryb oceniono na 0 pkt, po 24 godz. – 3 pkt, a po 48 godz. – 9 pkt. W tkance mięśniowej ryb stwierdzono istotny spadek pH (z 6,93 do 6,54) i wzrost przewodności elektrycznej właściwej (z 4,14 do 7,95 mS cmˉ¹). Instrumentalne pomiary właściwości fizykochemicznych potwierdziły zmianę świeżości określone indeksem (QIM), co wskazuje na praktyczne wykorzystanie tego wskaźnika dla ryb przechowywanych w warunkach chłodniczych w ocenianym okresie.

EN

The sensory characteristics of 30 rainbow trout during chilling storage for 48 hours using QIM system were evaluated. In addition, instrumental measurements of muscle intrinsic properties, i.e. value of pH and electrical conductivity, and color of skin (according to CIE L*a*b*) were determined. Quality Index (QI) after fish killing amounted 0 pts., after 24 h 3 pts., and after 48 h 9 pts. The significant pH decrease (from 6.93 to 6.54), and an electrical conductivity increase (from 4.14 to 7.95 mS cmˉ¹) in fish muscle tissue were found. Instrumental measurements of intrinsic properties which confirmed the change of freshness specified by QIM method, indicate the practical use of this index for fish stored in chilling conditions during evaluated period.

4

Preparation and anatomical distribution study of 67Ga-alginic acid nanoparticles for SPECT purposes in rainbow trout (Oncorhynchus mykiss)

Heidarieh M., Daryalal F., Mirvaghefi A., Rajabifar S., Diallo A., Sadeghi M., Zeiai F., Moodi S., Maadi E., Sheikhzadeh N., Heidarieh H., Hedyati M.

Nukleonika

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2014

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Vol. 59, No. 4

153--159

EN

Ergosan contains 1% alginic acid extracted from two brown sea weeds. Little is known about the target organs and anatomical distribution of Ergosan (alginic acid) in fi sh. Therefore, feasibility of developing alginic acid nanoparticles to detect target organ in rainbow trout is interesting. To make nanoparticles, Ergosan extract (alginic acid) was irradiated at 30 kGy in a cobalt-60 irradiator and characterized by transmission electron microscopy (TEM) and Fourier transform infrared spectroscopy (FTIR). Results from TEM images showed that particle sizes of irradiated alginic acid ranged from 30 to 70 nm. The FTIR results indicated that gamma irradiation had no signifi cant infl uence on the basic structure of alginic acid. Later, alginic acid nanoparticles were successively labelled with 67Ga-gallium chloride. The biodistribution of irradiated Ergosan in normal rainbow trout showed highest uptake in intestine and kidney and then in liver and kidney at 4- and 24-h post injection, respectively. Single-photon emission computed tomography (SPECT) images also demonstrated target specifi c binding of the tracer at 4- and 24-h post injection. In conclusion, the feed supplemented with alginic acid nanoparticles enhanced SPECT images of gastrointestinal morphology and immunity system in normal rainbow trout.

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New multiplex PCR assays for estimating genetic diversity in rainbow trout (Oncorhynchus mykiss) by polymorphism of microsatellite DNA

Kaczmarczyk D., Kaczor A.

Environmental Biotechnology

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2013

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Vol. 9, No. 1

19--24

EN

Multiplex PCR is a useful technique for estimating genetic diversity. This paper presents 3 new sets of primer pairs for effectively amplifying 10 microsatellite DNA loci from rainbow trout (Oncorhynchus mykiss). Unlike other sets of primer pairs that have been developed for amplifying rainbow trout microsatellite loci, ours do not require the hot-start PCR technique. In the paper, we describe the steps taken to choose the loci for each multiplex assay and to verify the genotyping results. We provide the compositions of the PCR mixture and the characteristics of the PCR thermal profile recommended for amplification.

6

Benzo[a]pyrene and cyclopenta[c]phenanthrene suppress expression of p53 in head kidney of rainbow trout

Brzuzan P., Woźny M., Góra M., Łuczyński M. K., Jabłońska A.

Environmental Biotechnology

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2011

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Vol. 7, No. 1

41-45

EN

Although p53, a protein of important tumor suppressive function, has been extensively studied in mammals, relatively little is known about the p53 pathways in lower vertebrates. Particularly, limited information exists on possible influences of environmental contaminants on the expression of the p53 gene in fish. In the current study, we assessed the effects of benzo[a]pyrene (B[a]P; potent tumor promoter) and cyclopenta[c]phenanthrene (CP[c]Ph; clastogenic agent) exposure on a 24h profile of p53 gene expression in head kidney of juvenile rainbow trout (Oncorhynchus mykiss). To analyze the p53 transcription rate, we developed protocol for the examination of both mRNA and heterogeneous nuclear (hn) RNA of the gene, using Real-Time RTPCR approach. The results show that both compounds are capable of suppressing p53 transcriptional activity within 12h of the treatment. Our finding supports the idea that structurally different PAHs may influence cell physiologic functions controlled by p53 in fish, in part, by down-regulating its RNA expression levels.

7

Alterations in selected haematological parameters of rainbow trout (Oncorhynchus mykiss) exposed to complex mixtures of contaminants

Vosyliene M. Z., Kazlauskiene A., Mikalajune A.

Environment Protection Engineering

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2010

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Vol. 36, nr 4

95-104

EN

The aim of the study was to evaluate and compare changes in the selected haematological parameters of rainbow trout after long-term (14 days) exposure to sublethal concentrations of two kinds of contaminants: 1) highway traffic emission pollutants (model mixture of heavy metals) and 2) road maintenance salt. Four groups of fish exposed to sublethal concentrations of model mixture metal solutions demonstrated an increased frequency of abnormal erythrocytes and elevated percentage of neutrophiles. The percentage of juvenile and abnormal erythrocytes was significantly increased in the blood of fish exposed to sublethal concentrations of road maintenance salt solutions. Our data confirm that even low amounts of metals found in soil near roadsides and road maintenance salt can induce adverse effects on fish. Morphological analysis of erythrocytes seems to be a reliable tool for detection of toxic effects upon fish.

8

CYP1A gene expression in adipose fin of rainbow trout (Oncorhynchus mykiss Walbaum) exposed to benzo[a]pyrene

Brzuzan P., Woźny M., Łuczyński M. K., Góra M.

Environmental Biotechnology

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2007

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Vol. 3, No. 1

20-24

EN

Proximate to the environment, adipose fin of fish may be considered as a lipid storing tissue, and thus can be a target for either waterborne or dietary polycyclic aromatic compounds (PACs). We determined the effects of benzo[a]pyrene (B[a]P), a model PAC member, on CYP1A gene expression in adipose fin and compared that with the effects in gill of juvenile rainbow trout (Oncorhynchus mykiss Walbaum) using the quantitative reverse transcription polymerase chain reaction (Q-RT-PCR). The results of the study demonstrated that constitutive CYP1A mRNA was present in adipose fin of rainbow trout, but the transcripts were far less abundant than those in gill tissue. We confirmed high CYP1A gene induction potential of the gills in rainbow trout injected with benzo[a]pyrene, but also showed moderately and transiently induced CYP1A mRNA in adipose fin. The modest and transitory gene expression may preclude rainbow trout adipose fin CYP1A mRNA levels from using it as an indicator of sustained exposure of fish to the polycyclic aromatic compounds.

9

Cyclopenta[c]phenanthrene induction of CYP1A in brain of rainbow trout (Oncorhynchus mykiss)

Brzuzan P., Woźny M., Łuczyński M. K., Góra M., Ciesielski S., Kuźmiński H.

Environmental Biotechnology

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2007

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Vol. 3, No. 1

10-14

EN

We assessed the effects of cyclopenta[c]phenanthrene (CP[c]Ph) and benzo[a]pyrene (B[a]P; positive control) on CYP1A gene expression in brain of juvenile rainbow trout (Oncorhynchus mykiss) using the quantitative reverse transcription polymerase chain reaction (Q-RT-PCR). A group of hatchery raised rainbow trout, with an average body mass of 49.4 g and total length of 15.5 cm were given an intraperitoneal injection (10 mg*kg-1) of either CP[c]Ph or B[a]P in corn oil (2 mg*mi-1 corn oil) or corn oil alone (control). After 24 and 48 h, trout brains were collected for mRNA isolation and analysis. After 24 hours of the exposure, only B[a]P-treated rainbow trout had 10-fold higher number of CYP1A transcripts (mean = 3.63*106 transcripts*µg-1 total RNA) than control fish (3.24*105 transcripts*µg-1 total RNA; Tukey test, P<0.05). After 48 hrs, significantly higher levels of CYP1A expression (Tukey test, P<0.001) were found in either CP[c]Ph- or B[a]P- induced group (1.45*106 and 6.92*106 transcriptsźµg-1 total RNA, respectively) over a control group (mean=1.41*105 transcripts*µg-1 total RNA). The finding that CYP1A in brain tissue was inducible by CP[c]Ph, a polycyclic aromatic hydrocarbon (PAH) of different than B[a]P planar characteristics, may further validate the use of rainbow trout brain CYP1A mRNA levels as a biomarker of PAH exposure.

10

Biotechnology in freshwater finfish aquaculture

Luczynski M., Ocalewicz K.

Environmental Biotechnology

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2007

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Vol. 3, No. 2

37-38