Liver Of Brown Trout Research Articles

A Step Forward in the Characterization of Primary Brown Trout Hepatocytic Spheroids as Experimental Models.

Mammal hepatocyte spheroids have been investigated as alternative experimental models in several contexts, since three-dimensional (3D) systems have shown the potential to mimic in vivo scenarios. The description of fish hepatocyte 3D models is still minimal. This study intends to further characterize brown trout primary hepatocyte spheroids at distinct time points up to 25 days in culture. Viability, biometry, histomorphology, and basal expression of a selection of genes (metabolism and detoxification, efflux transport, and estrogenic signalling) were considered. The gene expression of whole liver samples from the same fish donor were evaluated concurrently. After 12 days in culture, the hepatocyte spheroids exhibited biometric and morphological stability. From the 12th to the 20th day in culture, the basal expression levels for most of the selected genes did not vary. The targeted mRNA levels were higher in brown trout liver samples compared to hepatocyte spheroids. Despite that, data supported that this model resembles some in vivo features. As an experimental alternative model, it showed potential to be used in a stable time window that can be exploited for exposure tests to different xenobiotics, namely, estrogenic compounds.

Inter-individual variation in mitochondrial phosphorylation efficiency predicts growth rates in ectotherms at high temperatures.

There is increasing evidence that aquatic ectotherms are especially vulnerable to global warming since their metabolic demands increase with ambient temperature while water-oxygen content decreases. The possible role of shrinking aerobic scope in limiting performance has been much discussed; however, less attention has been given to whether tissue-level changes in the efficiency of oxygen usage occur at elevated temperatures. Here, we show that this varies widely among individuals, with consequences for performance. We examined the inter-individual variation in growth rate and mitochondrial function from white muscle and liver of brown trout (Salmo trutta) acclimated to either high (19.5°C) or near-optimal temperature (12°C). Liver (but not muscle) mitochondria showed a positive relationship between growth rate and maximal oxidative phosphorylation at both temperatures, and a negative relationship between growth rate and ROS release. There was a positive correlation in both tissues between individual mitochondrial phosphorylation efficiency and growth rate, but only at 19.5°C. In this representative of aquatic ectotherms, an individual's liver mitochondrial efficiency thus seems to dictate its capacity to grow at elevated temperatures. This suggests that individual heterogeneity in cellular function may cause variation in the thermal limits of aquatic ectotherms and could adversely affect wild populations in warming environments.

Biochemical and cellular biomarkers in brown trout (Salmo trutta f. fario) in response to the antidepressants citalopram and venlafaxine

BackgroundDuring the last decades, a worldwide increase in the number of cases of depression accompanied by rising prescription rates of antidepressants was recorded. In Germany, the two most prescribed antidepressants are the selective serotonin reuptake inhibitor (SSRI) citalopram and the serotonin and noradrenalin reuptake inhibitor (SNRI) venlafaxine, taking about 30% of the market share. Both antidepressants are found frequently in surface waters and have the potential of adversely affecting aquatic organisms. Most studies dealing with antidepressants address apical endpoints and behaviour; however, only few studies investigate biochemical biomarkers and histopathological alterations. We conducted citalopram and venlafaxine exposure experiments over 5 months, starting with brown trout eggs in the eyed-ova stage, as well as with juvenile brown trout for 4 weeks. Exposure concentrations ranged from environmentally relevant 1 µg/L up to 1 mg/L. In this study, we investigated the effects of the antidepressants on b-esterase activity (neurotoxicity), stress protein level (proteotoxicity) and superoxide dismutase activity (oxidative stress). Additionally, we assessed the health status of the liver by means of histopathological analyses.ResultsWe were able to show that both antidepressants did neither induce proteotoxic nor neurotoxic effects in brown trout. But for venlafaxine, the biochemical biomarker for oxidative stress (superoxide dismutase activity) was significantly increased in larvae exposed to at least 10-µg/L venlafaxine at 7 °C. With regard to liver histopathology, fish exposed to higher citalopram concentrations were in a worse health condition than control fish irrespective of their life stage. Also, the energy storage of fish exposed to 1 mg/L citalopram was reduced.ConclusionThus, we here report citalopram-dependent histopathological alterations in brown trout liver, and the induction of oxidative stress by venlafaxine.

Identification of a piscine reovirus-related pathogen in proliferative darkening syndrome (PDS) infected brown trout (Salmo trutta fario) using a next-generation technology detection pipeline.

The proliferative darkening syndrome (PDS) is an annually recurring disease that causes species-specific die-off of brown trout (Salmo trutta fario) with a mortality rate of near 100% in pre-alpine rivers of central Europe. So far the etiology and causation of this disease is still unclear. The objective of this study was to identify the cause of PDS using a next-generation technology detection pipeline. Following the hypothesis that PDS is caused by an infectious agent, brown trout specimens were exposed to water from a heavily affected pre-alpine river with annual occurrence of the disease. Specimens were sampled over the entire time period from potential infection through death. Transcriptomic analysis (microarray) and RT-qPCR of brown trout liver tissue evidenced strong gene expression response of immune-associated genes. Messenger RNA of specimens with synchronous immune expression profiles were ultra-deep sequenced using next-generation sequencing technology (NGS). Bioinformatic processing of generated reads and gap-filling Sanger re-sequencing of the identified pathogen genome revealed strong evidence that a piscine-related reovirus is the causative organism of PDS. The identified pathogen is phylogenetically closely related to the family of piscine reoviruses (PRV) which are considered as the causation of different fish diseases in Atlantic and Pacific salmonid species such as Salmo salar and Onchorhynchus kisutch. This study also highlights that the approach of first screening immune responses along a timeline in order to identify synchronously affected stages in different specimens which subsequently were ultra-deep sequenced is an effective approach in pathogen detection. In particular, the identification of specimens with synchronous molecular immune response patterns combined with NGS sequencing and gap-filling re-sequencing resulted in the successful pathogen detection of PDS.

Age- and sex-dependent variation in the activity of antioxidant enzymes in the brown trout (Salmo trutta)

Oxidative stress is defined as the imbalance between pro-oxidant and antioxidant molecules in favor of the former and it represents one of the main driving forces of aging. To counteract the harmful effects of oxidative stress, organisms evolved a complex antioxidant system. According to the free radical theory of aging, while the production of reactive oxygen species (ROS) increases with age, the antioxidant defenses decline. Although this relationship has been investigated in diverse vertebrate taxa, the information in fish is scant and inconsistent, particularly for wild populations. Thus, the aim of the present study was the investigation of age- and sex-related changes of the antioxidant enzymes activity in free-living individuals of the brown trout (Salmo trutta). We measured the activity of the main enzymes involved in antioxidant protection, namely superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and glutathione S-transferase (GST), as well as lipid peroxidation, in the gills and the liver dissected from brown trout (1+- to 5+-year-old). A significant age-dependent variation in the activity of antioxidant enzymes was noted, with the exception of CAT. GPx activity followed a significant increasing trend with age in both the organs, while SOD decreased in the liver. Increased GST activity was found in the gills only. Lipid peroxidation levels significantly decreased with age in both the organs. SOD and CAT showed sex-dependent differences in the liver of brown trout, with males showing lower enzymatic activity than females. Our data contribute to improve the knowledge on the relationship between antioxidant enzyme activity, aging, and sex in fish.

Cytosolic distributions of highly toxic metals Cd and Tl and several essential elements in the liver of brown trout (Salmo trutta L.) analyzed by size exclusion chromatography and inductively coupled plasma mass spectrometry

Cytosolic distributions of nonessential metals Cd and Tl and seven essential elements among compounds of different molecular masses were studied in the liver of brown trout (Salmo trutta) from the karstic Krka River in Croatia. Analyses were done by size exclusion high performance liquid chromatography and high resolution inductively coupled plasma mass spectrometry. Common feature of Cd and Tl, as highly toxic elements, was their distribution within only two narrow peaks. The increase of cytosolic Cd concentrations was reflected in marked increase of Cd elution within low molecular mass peak (maximum at ∼15 kDa), presumably containing metallothioneins (MTs), which indicated successful Cd detoxification in brown trout liver under studied exposure conditions. Contrary, the increase of cytosolic Tl concentrations was reflected in marked increase of Tl elution within high molecular mass peak (maximum at 140 kDa), which probably indicated incomplete Tl detoxification. Common feature of the majority of studied essential elements was their distribution within more peaks, often broad and not well resolved, which is consistent with their numerous physiological functions. Among observed associations of essential metals/nonmetal to proteins, the following could be singled out: Cu and Zn association to MTs, Fe association to storage protein ferritin, and Se association to compounds of very low molecular masses (<5 kDa). The obtained results present the first step towards identification of metal-binding compounds in hepatic cytosol of brown trout, and thus a significant contribution to better understanding of metal fate in the liver of that important bioindicator species.

Electrochemical Determination of Metallothioneins by the Modified Brdička Procedure as an Analytical Tool in Biomonitoring Studies

Present study aims to improve the modified Brdicka procedure of metallothionein (MT) quantification in a way to reduce the analysis duration. Therefore, scan rate of the voltammetric sweep was increased from 0.0052 V s–1 to 0.013 V s–1 by lowering the duration of one step potential from 0.5 s to 0.2 s, which resulted in 2.5 times faster voltammetric sweep. Research confirmed high accuracy and reliability of MT measurements, in both standard sample and samples of heat-treated cytosols of liver, gills and the intestine of European chub and liver of brown trout. The method was confirmed as fast and reliable electrochemical technique for quantification of MTs and this time- saving improvement is especially useful and applicable in different biomonitoring studies which require the analyses of numerous biological samples and high sample throughput.

Seasonal and Morphological Variations of Brown Trout (Salmo trutta f. fario) Kidney Peroxisomes: A Stereological Study.

Literature about fish kidney peroxisomes is scarce. To tackle this caveat, a stereological approach on renal peroxisome morphological parameters was performed for the first time in a fish, establishing correlations with maturation stages as it was previously done in brown trout liver. Three-year-old brown trout males and females were collected at the major seasons of their reproductive cycle. Trunk kidney was fixed and processed for catalase cytochemistry. Classical stereological methods were applied to electromicrographs to quantitate morphological parameters. Different seasonal variation patterns were observed between genders, and between renal proximal tubule segments I and II. In males, peroxisomes from proximal tubule segment II had a relatively higher volume and number in May, being individually bigger in February. Females presented similar trends, though with less marked variations. Overall, males and females did not show exactly the same seasonal patterns for most peroxisomal parameters, and no correlations were found between the latter and the gonado-somatic index (GSI). Hence, and despite the variations, the morphology of renal peroxisomes is not strictly correlated with gonad maturation kinetics, therefore suggesting that kidney peroxisome morphology is not seasonally modulated by sex steroids, like estradiol, as it seems to happen in liver peroxisomes.

Stereology of Brown Trout Liver Peroxisomes at Vitellogenesis and Pre-spawning Strengthens the Hypothesis of Their Regulation by Sex Steroids.

Stereology of Brown Trout Liver Peroxisomes at Vitellogenesis and Pre-spawning Strengthens the Hypothesis of Their Regulation by Sex Steroids.

Global transcriptomic profiling demonstrates induction of oxidative stress and of compensatory cellular stress responses in brown trout exposed to glyphosate and Roundup.

BackgroundGlyphosate, the active ingredient in Roundup formulations, is the most widely used herbicide worldwide, and as a result contaminates surface waters and has been detected in food residues, drinking water and human urine, raising concerns for potential environmental and human health impacts. Research has shown that glyphosate and Roundup can induce a broad range of biological effects in exposed organisms, particularly via generation of oxidative stress. However, there has been no comprehensive investigation of the global molecular mechanisms of toxicity of glyphosate and Roundup for any species. We aimed to characterise and compare the global mechanisms of toxicity of glyphosate and Roundup in the liver of brown trout (Salmo trutta), an ecologically and economically important vertebrate species, using RNA-seq on an Illumina HiSeq 2500 platform. To do this, we exposed juvenile female brown trout to 0, 0.01, 0.5 and 10 mg/L of glyphosate and Roundup (glyphosate acid equivalent) for 14 days, and sequenced 6 replicate liver samples from each treatment.ResultsWe assembled the brown trout transcriptome using an optimised de novo approach, and subsequent differential expression analysis identified a total of 1020 differentially-regulated transcripts across all treatments. These included transcripts encoding components of the antioxidant system, a number of stress-response proteins and pro-apoptotic signalling molecules. Functional analysis also revealed over-representation of pathways involved in regulating of cell-proliferation and turnover, and up-regulation of energy metabolism and other metabolic processes.ConclusionsThese transcriptional changes are consistent with generation of oxidative stress and the widespread induction of compensatory cellular stress response pathways. The mechanisms of toxicity identified were similar across both glyphosate and Roundup treatments, including for environmentally relevant concentrations. The significant alterations in transcript expression observed at the lowest concentrations tested raises concerns for the potential toxicity of this herbicide to fish populations inhabiting contaminated rivers.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-015-1254-5) contains supplementary material, which is available to authorized users.

Fate of Tetracapsuloides bryosalmonae (Myxozoa) after infection of brown trout Salmo trutta and rainbow trout Oncorhynchus mykiss

Tetracapsuloides bryosalmonae (Myxozoa) is the causative agent of proliferative kidney disease in salmonids. We assessed differences in intensity of T. bryosalmonae infection between brown trout Salmo trutta and rainbow trout Oncorhynchus mykiss from the clinical phase of infection onwards. Specific pathogen-free fish were exposed to T. bryosalmonae spores under controlled laboratory conditions and sampled at 6, 8, 10, 12, 14, and 17 wk post exposure (wpe), and the transmission of T. bryosalmonae from infected fish to the bryozoan Fredericella sultana was observed. Parasite load was determined in fish kidneys by quantitative real-time PCR (qRT-PCR), and parasite stages were detected in kidney, liver, and spleen tissues at different time points by immunohistochemistry. T. bryosalmonae was successfully transmitted from infected brown trout to F. sultana colonies but not from infected rainbow trout. Body length and weight of infected brown trout did not differ significantly from control brown trout during all time points, while length and weight of infected rainbow trout differed significantly compared to controls from 10 to 17 wpe. qRT-PCR revealed that parasite load was significantly higher in kidneys of brown trout compared with rainbow trout. Immunohistochemistry showed high numbers of intra-luminal stages (sporogonic stages) in kidneys of brown trout with low numbers of pre-sporogonic stages. Sporogonic stages were not seen in kidneys of rainbow trout; only high numbers of pre-sporogonic stages were detected. Numbers of pre-sporogonic stages were low in the spleen and liver of brown trout but high in rainbow trout. These data confirmed that there are differences in the development and infection progress of T. bryosalmonae between brown trout and rainbow trout.

Alterations in Fatty Acids of Polar Lipids in Salmo trutta on Long-term Exposure to a Glyphosate-Based Herbicide (Roundup®)

Abstract: In present study, the effects of sublethal doses (10 and 20 mg L(-1)) of Roundup on fatty acid pattern in muscle and liver of brown trout were investigated. For this purpose, fish were held in experiment tanks for 1 month. While total MUFA wasn't influenced, the highest total SFA and total n-6 PUFA were determined in group 10 mg L(-1) and the lowest values were determined in control group and group 20 mg L(-1) in muscle, respectively. The highest and the lowest total n-3 PUFA was found in control group and group of 10 mg L(-1) in muscle, respectively. Total n-3/n-6 PUFA ratio and EPA+DHA level of group 10 mg L(-1) were lower than other groups in muscle. The amount of total n-3/n-6 PUFA, EPA + DHA and total n-3 PUFA of control group were found higher than treatment groups in liver. While the highest total SFA was determined in group 10 mg L(-1), there wasn't difference between control group and group 20 mg L(-1) in liver. Both of doses herbicide had higher value than control for total MUFA in liver. While Roundup didn't inhibit n-3 PUFA synthesis in the muscle, both concentrations, exhibited inhibitory effect on n-3 PUFA synthesis in the liver. This result probably consequence of its indirect effect on the some enzyme activities or gene expressions in fatty acid metabolism of brown trout.

Pex11α in brown trout (Salmo trutta f. fario): Expression dynamics during the reproductive cycle reveals sex-specific seasonal patterns

A negative correlation between female gonadal maturation kinetics and size variations of hepatic peroxisomes was earlier documented in brown trout, as a probable impact of serum estrogen changes during the reproductive cycle. Herein, we investigated whether the organelle volume/surface dynamics seen in female brown trout liver peroxisomes – without numerical changes within each hepatocyte – is followed by variations in the expression of the membrane peroxisome protein Pex11α gene. For comparison, we also studied males. We find in females a seasonal variation with the highest Pex11α expression in February, which was statistically different from all other tested periods. Overall, the expression of PEX11α had over a fivefold decrease from February to September. This period coincides with the reproductive transition between the earlier post-spawning gonadal remodeling and preparatory staging and the pre-spawning period. Males did not show changes. Our approach allowed the first characterization of a peroxin gene in a teleost, the Pex11α, while offering a correlation scenario were, as we hypothesized, the peroxisomal size kinetics is paralleled by membrane-related gene alterations (measured herein as proxy of Pex11α gene expression). Our data support and expand previous results on the regulation, function and morphology of peroxisome dynamics in brown trout, with a broader interest.

The 17β-hydroxysteroid dehydrogenase 4: Gender-specific and seasonal gene expression in the liver of brown trout (Salmo trutta f. fario)

Previously, it was documented that liver peroxisomes display seasonal size changes in the adult Salmo trutta fario, especially in females (and negatively correlated with ovary maturation). It was then hypothesized that decreases in peroxisome size could be paralleled by changes in peroxisomal beta-oxidation and estradiol catabolism actions. The 17beta-hydroxysteroid dehydrogenase 4 has been portrayed as playing an important role in both processes. To elucidate its function in the described peroxisomal pattern, we isolated the cDNA and predicted the protein sequence of the enzyme in that species. The seasonal gene expression pattern in both genders was addressed through quantitative PCR. Fish sampling was in post-spawning period, early and advanced gonad maturation, and pre-spawning. Males did not vary seasonally. As to females, a seasonal pattern was evidenced according to our previous hypothesis. We suggest that the decreased levels observed during vitellogenesis are related to lipid needs for ovary maturation, and, additionally, with the need of modulating estradiol titers.

Seasonal and gender variation of peroxisome proliferator activated receptors expression in brown trout liver

PPAR isotypes have been previously identified in the teleost brown trout ( Salmo trutta f. fario) and their organ distribution pattern established. Being that the liver is a vital metabolic organ presenting expression of all isotypes and also knowing that estrogens/estrogen receptors seem to interact with PPARs, we hypothesized that the latter may very well change seasonally. So, we studied the expression of these receptors in the liver, along the annual reproductive cycle and in both genders. According to real-time RT-PCR, PPARα mRNA expression in females was significantly higher in May and lower in September than in other seasons. No significant variation was observed along the year in males. A significant difference between genders occurred in May, when PPARα expression was higher for females. PPARβ expression showed little variation along the reproductive cycle in females, but in males it was significantly higher in December than in the other seasons. No significant differences existed between genders. PPARγ was more expressed in February than in September and December, for females. As to males, it was more expressed in February than in all other seasons. No significant differences were observed between genders. The study proved our hypothesis that PPARs gene expression varies along the year. Moreover, PPARα expression in females followed the same annual variation pattern as peroxisome volumes and enzyme activities, and an inverse pattern relatively to the salmonid type annual plasma estradiol levels. The data agrees with the idea that PPARα is under estradiol modulation and that cross-talk between this receptor and the estrogen receptor possibly exists.

Heavy Metal Contamination of the Loučka River Water Ecosystem

Heavy metal contamination of the Loučka River water ecosystem was assessed in July 2005. We analyzed concentrations of T-Hg (total mercury), Cd, Pb, Cr, Cu, Zn, and Ni in water, sediments, zoobenthos, and in the brown trout (Salmo trutta m. fario) muscle and liver tissues (a total of 28 individuals) at four sampling sites. The highest Pb and Ni concentrations (4.634-12.050 and 0.689-24.980 mg kg-1) were found in sediments. The zoobenthos was most contaminated by Zn and Cu (0.556-1.505 and 2.925-74.300 mg kg-1). The heavy metal contamination of river water was highest in Ni and Cr (0.1-6.8 and 0.5-10.0 mg l-1). Concentrations of heavy metals in the brown trout muscle were following (in mg kg-1): Pb 0.108 ± 0.073 - 1.010 ± 0.506, Cd 0.003 ± 0.002 - 0.026 ± 0.022, Zn 3.956 ± 0.371 - 5.801 ± 1.718, Ni 0.058 ± 0.018 - 0.102 ± 0.046, Cr 0.028 ± 0.005 - 0.073 ± 0.039, Cu 0.329 ± 0.079 - 0.437 ± 0.064 and Hg 0.065 ± 0.008 - 0.106 ± 0.047. Statistical differences (P &lt; 0.05) in the brown trout muscle were in Pb and Zn. Cd, Cu and Zn were markedly accumulated in the brown trout liver (concentrations 0.107 ± 0.066 - 0.223 ± 0.078, 59.973 ± 38.951 - 145.800 ± 48.286 and 30.671 ± 3.574 - 34.274 ± 7.226 mg kg-1). Humans of 60 kg body mass may consume 1.5 kg of brown trout muscle from the Loučka River weekly without any risk. Adverse influence of the Uniglas distillery on the Loučka River environment contamination by heavy metals was not confirmed.

Strategies to Maximize Adhesion of Thick Paraffin Sections of the Brown Trout Liver for Stereological Purposes

Thick histological sections are essential for implementing many modern stereological techniques because most rely on the optical disector technique both for sampling and counting objects (namely cells). In conducting any stereological evaluation, objects must be unambiguously identified. In many instances, only immunocytochemistry allows the required clear cell identification in sections. In immunohistochemical procedures adhesion media are routinely used, especially when including aggressive treatments for antigen retrieval (eg, microwave exposure) and tissue permeabilization (eg, Triton X-100). Immunohistochemical techniques performed on thick sections (30–40 μm) can cause additional adhesion problems, reinforcing the need to seek the best adhesion strategies for particular situations.Formalin fixed liver of the brown trout was used in this study. After pre-treating the slides with 2N hydrochloric acid (HCl) and acetone, we compared 4 of the most used adhesion media, namely Mayer solution, poly-L-lysine, aminopropyltriethoxysilane (APES), and chromalumgelatin. A control procedure with a pre-treatment step was also used. For each adhesion medium and the control, 5 thick paraffin sections (40 μm) were used. After aggressive treatment with microwave irradiation and Triton X-100 and with an immunohistochemical avidin-biotin-like protocol, the sections were blindly and semiquantitatively evaluated. Only in treatments with APES and chromalum-gelatin was the adhesion satisfactory. APES was slightly more effective. A short illustration exercise using the single hepatocyte nucleus as the counting unit demonstrated the feasibility of the optical disector. This technique offers the best option because the chrormalum-gelatin costs less and there are only small differences between the 2 strategies. (The J Histotechnol 24:37, 2001)Submitted: September 6, 1999; Accepted with revisions: October 5, 2000

Strategies to Maximize Adhesion of Thick Paraffin Sections of the Brown Trout Liver for Stereological Purposes

Strategies to Maximize Adhesion of Thick Paraffin Sections of the Brown Trout Liver for Stereological Purposes

Brown trout Salmo trutta f. Fario liver ultrastructure as a biomarker for assessment of small stream pollution

T he effects of environmental pollutants from two small streams in south-west Germany on the liver of brown trout (S almo trutta f. Fario) were studied as biomarkers by means of quantitative and semi-quantitative electron microscopy, and quantitatively by morphometrical measurements. Cellular damage was assessed semi-quantitatively based on a classification of ultrastructural responses. Both methods revealed more severe cellular effects in the liver of trout which had been exposed to the highly polluted stream than in those exposed to the lightly polluted river. Morphometrical studies showed a significant reduction of glycogen storage and a significant increase in number of mitochondria, peroxisomes and cisternae of the endoplasmic reticulum. The biomarker responses of this study were correlated with the results obtained by limnological and analytical investigations, and reflect the levels of pollution in each stream.

Microanatomical organization of hepatic stroma of the brown trout, Salmo trutta fario (Teleostei, Salmonidae): A qualitative and quantitative approach.

Despite the absence of lobulation, light microscopy of serial sections of the liver of brown trout, Salmo trutta fario, reveals that the stromal elements are spatially organized as venous-biliary-arteriolar tracts (VBAT), venous-arteriolar tracts (VAT), biliary-arteriolar tracts (BAT), venous-biliary tracts (VBT), biliary tracts (BT), arteriolar tracts (AT), and isolated veins. These components are not two- but three-dimensional entities, and the anatomical interrelationships among all entities are displayed. The VBAT, VAT, and VBT are considered portal tracts; the adjacent parenchymal zones are viewed as periportal areas. The veins emerging from those tracts are regarded as afferent, and related with periportal zones. The veins that do not communicate with the VBAT, VAT, or VBT are viewed as efferent. Only serial sectioning allows a definite recognition of afferent from efferent isolated veins. The morphometric study discloses that isolated veins occupy around 60% of the stromal areas. Nevertheless, the VBAT, VAT, and BT are also considerably important, occupying variable proportions of the stromal areas (8-12%). The VBT and BAT are less important in quantitative terms. No sexual diffences appear in either qualitative or quantitative terms. There is no structural support for an eventual macroorganization of hepatic tissues. It is suggested that the quantitative data can be useful, as standards for the normal hepatic architecture of brown trout. The paper emphasizes the importance of a general structural model for the fish liver and of the use of an internationally acceptable nomenclature. © 1995 Wiley-Liss, Inc.