The myxozoan parasite Tetracapsuloides bryosalmonae is the causative agent of proliferative kidney disease (PKD)-a disease of salmonid fishes, notably of the commercially farmed rainbow trout Oncorhynchus mykiss. Both wild and farmed salmonids are threatened by this virulent/deadly disease, a chronic immunopathology characterized by massive lymphocyte proliferation and hyperplasia, which manifests as swollen kidneys in susceptible hosts. Studying the immune response towards the parasite helps us understand the causes and consequences of PKD. While examining the B cell population during a seasonal outbreak of PKD, we unexpectedly detected the B cell marker immunoglobulin M (IgM) on red blood cells (RBCs) of infected farmed rainbow trout. Here, we studied the nature of this IgM and this IgM+ cell population. We verified the presence of surface IgM via parallel approaches: flow cytometry, microscopy, and mass spectrometry. The levels of surface IgM (allowing complete resolution of IgM- RBCs from IgM+ RBCs) and frequency of IgM+ RBCs (with up to 99% of RBCs being positive) have not been described before in healthy fishes nor those suffering from disease. To assess the influence of the disease on these cells, we profiled the transcriptomes of teleost RBCs in health and disease. Compared to RBCs originating from healthy fish, PKD fundamentally altered RBCs in their metabolism, adhesion, and innate immune response to inflammation. In summary, RBCs play a larger role in host immunity than previously appreciated. Specifically, our findings indicate that the nucleated RBCs of rainbow trout interact with host IgM and contribute to the immune response in PKD.
- MeSH
- B-lymfocyty MeSH
- erytrocyty MeSH
- imunoglobulin M MeSH
- nemoci ledvin * MeSH
- Oncorhynchus mykiss * MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Salmincola markewitschi Shedko et Shedko, 2002 (Copepoda: Lernaeopodidae) is an ectoparasitic copepod mainly infecting the buccal cavities of white-spotted charr Salvelinus leucomaenis (Pallas) (Salmonidae). This species has only been recorded from Northeast Asia, where a morphologically similar congener Salmincola carpionis (Krøyer, 1837) is also distributed, using the same host species. These copepods are hard to distinguish from each other because of their similarities. We thus examined the newly collected specimens morphologically and genetically from five populations of white-spotted charr in Japan. Most of the specimens were morphologically consistent with S. markewitschi but showed great variations in the numbers of spines on the exopods of the antennae, shape of the maxilliped myxal palps, and the bulla diameter. Consequently, some specimens shared characteristics with S. carpionis. In addition to the mophological continuities, genetic analyses of 28S rDNA and COI mitochondrial DNA confirmed that all specimens belong to a single species. Further taxonomic revisions are required to draw conclusions of whether S. markewitschi is a valid species different from S. carpionis, by collecting samples from across their wide distributional ranges, such as Europe, North America, and Northeast Asia. A key to identification of species of Salmincola Wilson, 1915 occurring in Japan is also provided.
- MeSH
- Copepoda * genetika MeSH
- hostitelská specificita MeSH
- pstruh genetika parazitologie MeSH
- ribozomální DNA MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Evropa MeSH
One of the main contributors to pharmaceutical pollution of surface waters are non-steroidal anti-inflammatory drugs (NSAIDs) that contaminate the food chain and affect non-target water species. As there are not many studies focusing on toxic effects of NSAIDs on freshwater fish species and specially effects after dietary exposure, we selected rainbow trout (Oncorhynchus mykiss) as the ideal model to examine the impact of two NSAIDs - diclofenac (DCF) and ibuprofen (IBP). The aim of our study was to test toxicity of environmentally relevant concentrations of these drugs together with exposure doses of 100× higher, including their mixture; and to deepen knowledge about the mechanism of toxicity of these drugs. This study revealed kidneys as the most affected organ with hyalinosis, an increase in oxidative stress markers, and changes in gene expression of heat shock protein 70 to be signs of renal toxicity. Furthermore, hepatotoxicity was confirmed by histopathological analysis (i.e. dystrophy, congestion, and inflammatory cell increase), change in biochemical markers, increase in heat shock protein 70 mRNA, and by oxidative stress analysis. The gills were locally deformed and showed signs of inflammatory processes and necrotic areas. Given the increase in oxidative stress markers and heat shock protein 70 mRNA, severe impairment of oxygen transport may be one of the toxic pathways of NSAIDs. Regarding the microbiota, an overgrowth of Gram-positive species was detected; in particular, significant dysbiosis in the Fusobacteria/Firmicutes ratio was observed. In conclusion, the changes observed after dietary exposure to NSAIDs can influence the organism homeostasis, induce ROS production, potentiate inflammations, and cause gut dysbiosis. Even the environmentally relevant concentration of NSAIDs pose a risk to the aquatic ecosystem as it changed O. mykiss health parameters and we assume that the toxicity of NSAIDs manifests itself at the level of mitochondria and proteins.
- MeSH
- antiflogistika nesteroidní metabolismus MeSH
- biologické markery metabolismus MeSH
- chemické látky znečišťující vodu * metabolismus MeSH
- diklofenak metabolismus MeSH
- dysbióza MeSH
- ekosystém MeSH
- epidemický výskyt choroby MeSH
- ibuprofen metabolismus toxicita MeSH
- kyslík metabolismus MeSH
- léčivé přípravky metabolismus MeSH
- messenger RNA metabolismus MeSH
- Oncorhynchus mykiss * metabolismus MeSH
- oxidační stres MeSH
- proteiny tepelného šoku HSP70 metabolismus MeSH
- reaktivní formy kyslíku metabolismus MeSH
- střevní mikroflóra * MeSH
- voda metabolismus MeSH
- zánět chemicky indukované MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Výskum probiotík pre akvakultúru je v ranom štádiu a pre ich implementáciu je potrebné vykonať ešte množstvo experimentov. Laktiplantibacily patria medzi mikroorganizmy, ktoré sa najčastejšie používajú na prípravu probiotických preparátov. Doterajšie výsledky nie sú postačujúce, práve preto sú potrebné ďalšie štúdie. Výber probiotík pre akvakultúru a ich vývoj pre komerčné využitie v akvakultúre je mnohostupňový a multidisciplinárny proces vyžadujúci si v prvej etape základný a neskôr aj aplikovaný výskum a posúdenie jeho použitia v praxi. Cieľom štúdie bolo pripraviť probiotické krmivo pre ryby s využitím pomocných látok a následne sledovať prežívateľnosť probiotických bakteriálnych buniek v krmive počas 9-mesačného skladovania pri chladničkovej (4 °C) a izbovej teplote (22 °C). Na prípravu krmiva bol použitý kmeň Lactobacillus plantarum R2 Biocenol™ (CCM 8674) (podľa novej taxonómie Lactiplantibacillus plantarum), potenciálne využiteľný pre probiotické účely v akvakultúre. Lepšia prežívateľnosť probiotických bakteriálnych buniek bola zaznamenaná vo vzorkách krmiva A (Aquatex 41 HMD) v porovnaní so vzorkami probiotických peliet B (Inicio 918-2). Keďže oxidácia mastných kyselín v krmive ovplyvňuje nutričnú kvalitu jednotlivých komponentov krmiva, predpokladáme, že vyššie množstvo oleja v krmive B negatívne ovplyvnilo prežívateľnosť probiotických bakteriálnych buniek. Najvyššie počty životaschopných probiotických baktérií boli zaznamenané pri 4 °C skladovania krmiva. Po 9 mesiacoch skladovania pri chladničkovej teplote počty laktiplantibacilov vo vzorkách krmiva A klesli z hodnoty 7,30 log10KTJ/g na počet 5,57 log10KTJ/g. Teplota je považovaná za rozhodujúci faktor ovplyvňujúci životaschopnosť a prežívateľnosť probiotických baktérií počas doby skladovania.
Research in probiotics for aquaculture is at an early stage of development and much work is still needed. Lactiplantibacilli belong to the microorganisms most frequently used to prepare the probiotics. The available information is inconclusive, since few experiments with sufficiently robust design have been conducted to permit critical evaluation. The development of probiotics applicable to commercial use in aquaculture is a multistep and multidisciplinary process requiring both empirical and fundamental research, full-scale trials, and an economic assessment of its use. The aim of the study was to prepare a probiotic aquafeed via excipients and subsequently to observe the survival of probiotic bacterial cells in the feed during the nine months storage period at a refrigerator (4 °C) or room temperature (22 °C). The strain Lactobacillus plantarum R2 Biocenol™ (CCM 8674) (according to the new taxonomy Lactiplantibacillus plantarum), potentially usable as a probiotic in aquaculture, was administered to prepare the aquafeed. Better survival of probiotic bacterial cells was recorded in a samples of pellets A (Aquatex 41 HMD) compared to the samples of probiotic pellets B (Inicio 918-2). Since oxidation of fatty acids in feed affects the nutritional quality of individual feed components, we assume that higher amounts of oil in feed B negatively affected the survival of probiotic bacterial cells. The highest numbers of viable probiotic bacteria cells were recorded at 4 °C storage of probiotic feed samples. The number of lactiplantibacilli dropped from 7.30 log10CFU . g–1 to 5.57 log10CFU . g–1 after the nine months storage period of feed samples A at 4 °C. Temperature is considered as a critical factor influencing probiotic viability and survival during storage period.
The aim of this study was to reveal the effects of foodborne fluoxetine on morphological and condition profile, hematological profile, biochemical and oxidative stress indices on juvenile rainbow trout. The study was performed according to OECD Guideline No. 215. Fluoxetine was incorporated into Biomar 921 pellets at a dose of 0.047 mg/kg (environmental concentration), 0.577 mg/kg and 6.7 mg/kg. There was statistically significant change in hematological profile, including an increasing trend in neutrophil/lymphocyte ratio and a decreasing trend in the number of lymphocytes. Measurements of oxidative stress indicated decreased activity of the detoxifying enzyme glutathione-S-transferase in the liver and kidney. However, the measurement of GR, GPx, CAT, SOD activity, and TBARS showed no changes. Histopathological examination revealed damage to the proximal tubules of caudal kidney in exposed groups. This study confirms that fluoxetine has a significant effect on immune response.
- MeSH
- antidepresiva druhé generace toxicita MeSH
- chemické látky znečišťující vodu toxicita MeSH
- fluoxetin toxicita MeSH
- imunita účinky léků MeSH
- kontaminace potravin MeSH
- krevní obraz MeSH
- krmivo pro zvířata MeSH
- Oncorhynchus mykiss krev imunologie MeSH
- oxidační stres účinky léků MeSH
- proximální tubuly ledvin účinky léků MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Two genotypes of the intestinal parasite Ceratonova shasta infect Oncorhynchus mykiss: genotype 0 results in a chronic infection with low mortality while genotype IIR causes disease with high mortality. We determined parasite load and the relative expression of six immune factors (IgT, IgM, IL-6, IL-8, IL-10, IFNG) in fish infected with either genotype over 29 days post-exposure. In genotype IIR infections the host responded with upregulation of inflammatory and regulatory cytokines. In contrast, genotype 0 infection did not elicit an inflammatory response and expression of IFNG and IL-10 was lower. Antibody expression was upregulated in both infections but appeared to have limited efficacy in the virulent genotype IIR infections. Histologically, in genotype 0 infections the parasite migrated through the tissue layers causing inflammation but minimal damage to the mucosal epithelium, which contrasts with the severe pathology found in genotype IIR infections.
- MeSH
- cytokiny genetika metabolismus MeSH
- genotyp * MeSH
- imunoglobulin M krev MeSH
- imunoglobuliny krev MeSH
- interakce hostitele a parazita MeSH
- Myxozoa genetika patogenita MeSH
- nemoci ryb imunologie MeSH
- Oncorhynchus mykiss imunologie MeSH
- parazitární nemoci u zvířat imunologie MeSH
- parazitární zátěž MeSH
- pohyb buněk MeSH
- rybí proteiny krev MeSH
- sliznice imunologie MeSH
- virulence MeSH
- zánět imunologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
Proliferative kidney disease (PKD) is a widespread temperature-dependent disease in salmonids caused by the myxozoan parasite, Tetracapsuloides bryosalmonae (Canning, Curry, Feist, Longshaw et Okamura, 1999) (Tb). Tb has a two-host life cycle, involving fish as an intermediate host and freshwater bryozoans as the definitive host. Although salmonids are acknowledged as hosts for the parasite, it is less clear which fish species are active hosts in the life cycle of Tb. Differences in infection dynamics have been observed between some fish species, which are thought to be related to the existence of two main Tb-strains, the American and European. Iceland, having three species of indigenous salmonids and positioned geographically between Europe and North America, is an ideal location to study the natural development of Tb in wild fish. The main aim of this study was to determine the genetic origin of Tb in Iceland and confirm whether mature spores are produced in Icelandic salmonids. In this study, Icelandic salmonids were infected with the European Tb-strain. In situ hybridisation revealed that intraluminal sporogonic stages, including mature spores, were commonly observed in all three salmonid species. The presence of intraluminal stages has previously been confirmed in brown trout Salmo trutta Linnaeus and Atlantic salmon S. salar Linnaeus in Europe, but they have only been observed in Arctic charr Salvelinus alpinus (Linnaeus) in North America, infected by the local strain. This is, therefore, the first time that sporogonic stages have been observed in Arctic charr in Europe, where fish are infected with the European Tb-strain. Our data strongly suggest that all the three salmonid species inhabiting Icelandic waters serve as active hosts in the life cycle of Tb. However, for full confirmation, transmission trials are needed.
- MeSH
- hybridizace in situ veterinární MeSH
- interakce hostitele a parazita * MeSH
- Myxozoa růst a vývoj fyziologie MeSH
- nemoci ryb parazitologie MeSH
- parazitární nemoci u zvířat parazitologie MeSH
- pstruh * MeSH
- Salmo salar * MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Due to the fact that plastic pollution is a global environmental problem of modern age, studies on the impact of these synthetic materials on aquatic, and especially fish organisms, are an important part of the ecosystem and human nutrition. In our study, the toxicity of pristine polyethylene (PE) microparticles (approx. 50 μm) on rainbow trout (Oncorhynchus mykiss) was tested in three different dietary concentrations - 0.5%, 2% and 5%. After six weeks of exposure, various health indices were evaluated. Electron microscopy of the intestine revealed the disintegration of PE particles to <5 μm in size, and thus we concluded that microplastics are able to reach tissues. The haematological profile revealed changes in total red blood cells count and haematocrit (5% PE) which could be associated with spleen congestion observed histologically. The marker of lipid peroxidation was increased in gills suggesting the disruption of balance in antioxidant enzymes capacity and histopathological imaging revealed inflammation in higher PE concentrations. In addition, ammonia was decreased and calcium elevated in biochemical profile, confirming the gill damage. Electron microscopy of the gills showed lesions of lamellae and visible rings around the mucinous cell opening indicating their higher activity. Another injured was the liver tissue, as confirmed by hepatodystrophies and increased expression of pro-inflammatory genes in 2% PE. Impaired innate immunity was confirmed by an increased presence of mucinous cells and a decrease in leukocytes. Kidney damage manifested itself by higher expression of pro-inflammatory cytokines and histopathology. The damage in gills, liver and kidney together correlated with the increased antioxidant capacity of plasma. In conclusion, PE microparticles are able to affect health indices of O. mykiss. The potential problem for aquatic ecosystems and even human consumption should be considered.
Piscine cytochrome P450 (CYP) enzymes play an important role in the metabolism of xenobiotics. Xenobiotics often act as inducers of CYP1A1 and CYP3A expression and activity in fish. We compared constitutive mRNA expression of CYP1A1, CYP3A27, and CYP3A45 and catalytic activity of CYP1A (7-ethoxyresorufin-O-deethylation, EROD) and CYP3A-like (benzyloxy-4-trifluoromethylcoumarin-O-debenzyloxylation, BFCOD) enzymes in the following six rainbow trout tissues: liver, gill, heart, brain, intestine, and gonad. mRNA expression and activity were present in all investigated tissues. The CYP1A1 mRNA expression was higher in the liver, gill, heart, and brain compared to gonad and intestine. The intestine was the main site of CYP3A27 and CYP3A45 expression. The highest EROD and BFCOD activity was observed in liver tissue followed in descending order by heart, brain, gill, intestine, and gonad. Such differences might be related to the role of CYP physiological functions in the specific tissue. Rainbow trout exposure to 50 mg/kg of β-naphthoflavone for 48 h resulted in a 7.5- and 5.9-fold increase in liver EROD and BFCOD activity, respectively. In vitro EROD activity inhibition with ellipticine showed tissue-specific inhibition, while ketoconazole decreased BFCOD activity by 50-98 % in all tissues. Further studies are needed to identify all CYP isoforms that are responsible for these activities and modes of regulation.
- MeSH
- cytochrom P-450 CYP1A1 genetika metabolismus MeSH
- cytochrom P-450 CYP3A genetika metabolismus MeSH
- játra enzymologie MeSH
- messenger RNA genetika metabolismus MeSH
- mozek enzymologie MeSH
- myokard enzymologie MeSH
- Oncorhynchus mykiss metabolismus MeSH
- pohlavní dimorfismus MeSH
- regulace genové exprese enzymů fyziologie MeSH
- střeva enzymologie MeSH
- žábry enzymologie MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
The growing consumption of pharmaceuticals in the human population and the insufficient efficiency of their elimination in waste water has a long-term negative impact on the environment of aquatic ecosystems, including the organisms that inhabit them. A significant contributor is the consumption of anti-depressants from the SSRI group, which corresponds to their increasing concentration in the environment. The aim of this work was to determine if antidepressant sertraline is able to be stored in fish organisms and to evaluate the content of residues in various body tissues. Rainbow trout (Oncorhynchuss mykkis) was selected as the test organism and was artificially exposed to the antidepressant for 1 month (concentrations 0; 4.2; 44 and 400 ng.g-1 sertraline in the feed). Liver, kidney, brain and muscle tissue biopsies samples were taken for analysis. Detection was performed using an Accela 1250 LC pump and an Accela autosampler coupled with a high-performance mass analyzer with a heated electrospray ionization source Q-Exactive Orbitrap, operating in positive ionization mode and in PRM mode (m/z 306.08108->275.03888 and 309.009991->275.03888 for sertraline and internal standard, respectively). The limit of quantification of the method was 0.1 ng.g-1 of sertraline and the calibration curve showed a good linearity up to 20 ng.g-1. From the collected data, amount of residues was found in the liver, kidney and brain. In contrast, the incidence of residues in muscle tissue was not detected in all groups, which is favorable from the point of view of fish meat consumption, by humans.
- MeSH
- chemické látky znečišťující vodu analýza MeSH
- hmotnostní spektrometrie s elektrosprejovou ionizací * MeSH
- obsah radioaktivních látek v organizmu MeSH
- Oncorhynchus mykiss metabolismus MeSH
- rezidua léčiv analýza MeSH
- selektivní inhibitory zpětného vychytávání serotoninu analýza MeSH
- sertralin analýza MeSH
- tandemová hmotnostní spektrometrie * MeSH
- tkáňová distribuce MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
