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Limnoperna fortunei, the golden mussel, is a bivalve mollusk considered an invader in South America. This species is responsible for ecological and economic damages due to its voluminous fouling capability. Chemical biocides such as MXD-100™ and sodium dichloroisocyanurate (NaDCC) are often used to control L. fortunei infestations in hydraulic systems. Thus, we proposed to investigate the effects of different periods (24, 48 and 72 h) of exposure to MXD-100™ (0.56 mg L−1) and NaDCC (1.5 mg L−1) on the gills of L. fortunei through morphological and molecular analyses. NaDCC promoted progressive morphological changes during the analyzed periods and only an upregulation of SOD and HSP70 expression during the first 24 h of exposure. MXD-100™ led to severe morphological changes from the first period of exposure, in addition to an upregulation of SOD, CAT, HSP70 and CYP expression during the first 24 h. In contrast, MXD-100™ led to a downregulation of CAT transcription between 24 and 48 h. In static conditions, NaDCC causes lethal damage after 72 h of exposure, and that exposure needs to be continuous to achieve the control of the species. Meanwhile, the MXD-100™ treatment presented several effects during the first 24 h, showing acute toxicity in a shorter period of time.

This study identified the cellular compositions of the gills in molly fish and their role in immunity using light-, electron- microscopy, and immunohistochemistry. The molly fish gills consisted of four holobranchs spaced between five branchial slits. Each hemibranch carried many fine primary and secondary gill lamellae. The gill arch was a curved cartilaginous structure, from which radiated the bony supports of the primary lamellae. The gill arch contained the afferent and efferent brachial arteries. The gill arch was covered by epidermal tissue rich with mucous cells. The primary lamella had a central cartilaginous support and efferent and afferent arterioles and was covered with pavement cells (PVC), salt-secreting chloride cells, and pale-staining mucous cells. These chloride cells contained abundant mitochondria and tubulovesicular system and are involved in ionic transport with a potential role in detoxification. The surface of the secondary lamellae (site of gaseous exchange) consisted of overlapping or interdigitating PVC supported and separated by pillar cells. Other cells were found within the gill epithelium and interstitial connective tissues, including lymphocytes, macrophages, monocytes, telocytes, stem cells, astrocytes, and neuroepithelial cells. The immunohistochemical analysis revealed that APG-5, iNOS-2, IL-1β, NF-κB, and TGF-B showed positive immunoreactivity in macrophages. The epithelium of the primary gill lamellae contained positive-GFAP astrocytes and S100 protein—chloride cells. The stem cells expressed SOX9, myostatin, and Nrf2. Neuroendocrine cells expressed S100 protein. In conclusion, the current work suggests that the gills of molly fish are multifunctional organs and are involved in immune reactions.

The Ukrainian brook lamprey Eudontomyzon mariae is the most widespread lamprey species in eastern Europe. Although E. mariae is generally considered a derivative of Eudontomyzon danfordi , an exclusively freshwater parasitic species, it has alternatively been suggested that it was recently derived from a now extinct anadromous Black Sea ancestor. Several non-parasitic lampreys and the landlocked sea lamprey, which have recently evolved from anadromous ancestors, still develop a seawater-type mitochondria-rich cell (SW-MRC) in their gills. In contrast, this cell type is not present in the gills of either Lampetra aepyptera , a non-parasitic lamprey of ancient origin, or the parasitic Ichthyomyzon unicuspis and I. castaneus that likewise have long evolutionary histories in fresh water. Eudontomyzon mariae from the Vistula River in the Baltic River basin does not possess SW-MRC, which is inconsistent with a recent origin from an anadromous ancestor. Mitochondrial DNA sequence data were thus used to infer the relationship between different populations of E. mariae and E. danfordi , and to reconstruct the transition from anadromy to freshwater residency. The results suggest that E. mariae evolved independently in the Baltic, Black, and Caspian Sea basins, and not recently from an anadromous ancestor. Although E. mariae in the Danube River may have arisen relatively recently from E. danfordi (differing by 0.7–1.1% in cytochrome b gene sequence), other E. mariae populations (including in the Vistula River) are genetically closer (0.6%) to the hypothetical ancestor of both E. mariae and E. danfordi . That ancestor was probably a freshwater resident, since SW-MRCs are not rapidly lost following confinement in fresh water.

This study is an attempt to go further in the comprehension of the effects of heavy fuel oil in the context of an accidental oil spill at sea. It focuses on the link between morphological and functional impacts of realistic doses of the dissolved fraction of a heavy fuel oil on fish gills. Juvenile turbot, Scophthalmus maximus were exposed to the dissolved fraction of a heavy fuel oil for 5 days and then placed 30 days in clean sea water for recovery. During the contamination period, the concentration of the 16 US EPA priority poly-aromatic hydrocarbons showed small variations around a mean value of 321.0 ± 9.1 ng l −1 (mean ± SEM). The contamination induced a 64% increase in hepatic cytochrome P 450 1A (Western blot analysis). Osmolality, [Na + ] and [Cl − ] rapidly and significantly increased (by 14, 23 and 28% respectively) and slowly decreased to normal levels during the recovery period. At the same time, branchial histology showed decreases in the number of mucocytes (by 30%) and of chloride cells (by 95%) in the interlamellar epithelium. Therefore, it is suggested that the osmotic imbalance observed after the 5 days of exposure to the dissolved fraction of the heavy fuel oil is the consequence of the structural alteration of the gills i.e, the strong reduction of ionocyte numbers.

For the first time by the example of juveniles of Russian sturgeon Acipenser gueldenstaedtii Brandt, the role of growth hormone of hypophysis in the osmotic regulation of acipenserids was studied, also the process of transformation of reserve chloride was described. In Russian sturgeon juveniles, administration of growth hormone of carp Cyprinus carpio at a dose of 1 mg per 1 g body weight causes changes in the fine structure of numerous reserve (undifferentiated) gill epithelial cells and thus stimulates their transformation into mature chloride cells involved in transport of monovalent ions. Increase in the number of chloride cells, on account of reserve cells, was accompanied by higher enzymatic activity of Na + /K + -ATPase in gill homogenates of the injected fish adapting to a hyperosmotic environment of salinity 12.5‰ (403 mosm/l) than in the intact fish under the same conditions but without hormonal treatment. Morphophysiological changes observed in the injected juveniles provided the more efficient dynamics of blood serum osmolarity under saline exposure than that in the intact fish. Their serum osmolality during the maximum rise (24 hours after the transfer of fish into the water of 12.5‰ salinity) was lower ( p <0.01), than that of the intact fish. The inclusion of growth hormone in the osmoregulatory process was established.

Most teleost fish are ammoniotelic, and relatively few are ureotelic, in which the majority of nitrogenous waste is excreted as urea. This study aimed to determine whether the gill ultrastructure of ureotelic fish might have specific, unique characteristics compared with ammoniotelic fish. The gill morphology was studied in three closely related species of the family Batrachoididae: Opsanus beta, the gulf toadfish; Opsanus tau, the oyster toadfish; and Porichthys notatus, the plainfin midshipman, because prior studies have demonstrated that the two former species are ureotelic and excrete urea in unique, short daily pulses, whereas the latter is ammoniotelic. Ultrastructural studies demonstrated significant trafficking of dense-cored vesicles (50-200 nm) between the Golgi apparatus and the apical membrane of epithelial cells surrounding gill filaments and lamellae in these two Opsanus spp. The material constituting the core of these vesicles was intensely stained by lead salt and was unloaded externally when vesicles contacted the apical membrane. Another characteristic of these urea-secreting fish was the presence of numerous large, black-stained lysosomes, which contained cored vesicles, suggesting a second destination for the dense-cored vesicles. As a working hypothesis, the present data suggest that the urea-transporter protein, recently found in toadfish gills, is inserted in the vesicle. Subsequently, it could serve to either sequester cytosolic urea that ultimately is secreted into the water after contact of these vesicles with the pavement cell apical membrane, or it could allow facilitated diffusion of urea across the plasma membrane following insertion into the membrane. As further comparative evidence, the ammoniotelic P. notatus exhibited neither the vesicular trafficking nor the population of lysosomes both found in Opsanus spp.

Pesticides play a crucial role in agriculture, public health management, and veterinary practice to control pests. However, the persistent use of pesticides poses a significant threat to aquatic ecosystems and non-target organisms, such as fish. The genotoxicity, oxidative stress, and erythrocytic toxicity were assessed in fish exposed to different sublethal concentrations (5, 10, and 15 µg/L) of thiram for 12 days. Blood and other tissues (gills and intestine) were obtained from each fish reared in each group at days 4, 8, and 12 of the experiment. Thin blood film/smear examination indicated significantly ( P ≤ 0.05 ) increased morphological and nuclear changes in erythrocytes, such as the formation of micronuclei, notched nuclei, lobed nuclei, and tear-shaped erythrocytes in fish exposed to higher doses of thiram. The contents of antioxidant enzymes such as catalase (CAT), reduced glutathione (GSH), peroxidase (POD), and superoxide dismutase (SOD) were significantly ( P ≤ 0.05 ) decreased and the oxidative stress biomarkers including reactive oxygen species (ROS) and thiobarbituric acid reactive substances (TBARS) were significantly ( P ≤ 0.05 ) increased in gills and intestinal tissues of exposed fish. The results on genotoxicity determined using the comet assay technique revealed significantly ( P ≤ 0.05 ) high levels of genomic instability in terms of DNA damage in isolated cells of the gills and intestine of fish reared in water containing higher concentrations of thiram. At the microscopic level, various sections of the gills of fish treated with higher doses of thiram unveiled moderate to severe histopathological ailments such as degeneration of cartilaginous core, degeneration and disruption of primary and secondary lamellae of gills, necrosis of lamellar epithelial cells, telangiectasia, and curling of secondary lamellae. The findings of this study indicated that extremely low doses of thiram lead to the induction of adverse effects in freshwater fish. Hence, there is a need to develop suitable control measures regarding the use of environmentally friendly chemicals and for the protection of aquatic environments.

In fish, the gill plays a vital role in regulating the absorption of trace metals and is also highly susceptible to metal toxicity. Trace metals such as iron (Fe), copper (Cu), zinc (Zn), and manganese (Mn) are involved in various catalytic activities and molecular binding within the gill, thereby supporting a range of physiological processes in this organ. While beneficial at normal levels, these metals can become toxic when present in excess. Conversely, nonessential metals like cadmium (Cd) and lead (Pb) can gain entry into gill cells through similar metal transport pathways, potentially interfering with various cellular processes. The transepithelial transport of these metals across the gill epithelium is governed by a variety of metal transport and metal binding proteins. These include the Cu transporter 1 (CTR1), divalent metal transporter 1 (DMT1), and members of the Zrt-/Irt-like protein (ZIP) and zinc transport (ZnT) families. Additionally, some of these metals can compete with major ions (e.g., calcium, sodium) for absorption sites in the gill. This complex crosstalk suggests an interdependent mechanism that balances metal uptake to meet physiological needs while preventing excessive accumulation. In this article, I review the roles of trace metals in proteins/enzymes that support the different functions in the gill of teleost fish. I also discuss current understanding of the pathways involved in regulating the branchial uptake of metals and their influence on ionic regulation, and the potential detoxification mechanisms in the gill. Finally, I summarize knowledge gaps and potential areas for further investigation.

Rainbow trout ( Oncorhynchus mykiss ) are frequently used as experimental animals in ecotoxicological studies, in which they are experimentally exposed to defined concentrations of test substances, such as heavy metals, pesticides, or pharmaceuticals. Following exposure to a broad variety of aquatic pollutants, early morphologically detectable toxic effects often manifest in alterations of the gills. Suitable methods for an accurate and unbiased quantitative characterization of the type and the extent of morphological gill alterations are therefore essential prerequisites for recognition, objective evaluation and comparison of the severity of gill lesions. The aim of the present guidelines is to provide practicable, standardized and detailed protocols for the application of unbiased quantitative stereological analyses of relevant morphological parameters of the gills of rainbow trout. These gill parameters inter alia include the total volume of the primary and secondary gill lamellae, the surface area of the secondary gill lamellae epithelium ( i . e ., the respiratory surface) and the thickness of the diffusion barrier. The featured protocols are adapted to fish of frequently used body size classes (300–2000 g). They include well-established, conventional sampling methods, probes and test systems for unbiased quantitative stereological analyses of light- and electron microscopic 2-D gill sections, as well as the application of modern 3-D light sheet fluorescence microscopy (LSFM) of optically cleared gill samples as an innovative, fast and efficient quantitative morphological analysis approach. The methods shown here provide a basis for standardized and representative state-of-the-art quantitative morphological analyses of trout gills, ensuring the unbiasedness and reproducibility, as well as the intra- and inter-study comparability of analyses results. Their broad implementation will therefore significantly contribute to the reliable identification of no observed effect concentration (NOEC) limits in ecotoxicological studies and, moreover, to limit the number of experimental animals by reduction of unnecessary repetition of experiments.