Gill Filaments Research Articles

Effects of chlorpyrifos on histopathological biomarkers of the freshwater teleost Oreochromis niloticus

Abstract An attempt was made to evaluate histopathological changes in the gills, liver, and kidney tissues of the freshwater teleost Oreochromis niloticus as biomarkers of chlorpyrifos toxicity. An experiment was conducted in glass aquaria with O. niloticus exposed for 28 days to sub-lethal concentrations of chlorpyrifos of 0.0 µg L−1 (control), 10 µg L−1, and 20 µg L−1. Chlorpyrifos caused distortions of the primary gill lamellae structure, the curling of secondary lamellae, desquamation, and epithelial hyperplasia on secondary gill filaments. Hepatocyte vacuolation and nuclear membrane necrosis were found in the liver. Additionally, the shrinkage of the glomeruli, thickening of Bowman’s capsule, glomerular and tubular necrosis, intracytoplasmic vacuoles, and hyaline degeneration in the renal tubule were seen in the kidney of O. niloticus exposed to chlorpyrifos. No remarkable lesions were observed in the control group. The severity of lesions in the fish tissues was assessed using the standard semi-quantitative grading system. The degree of histopathological lesions in various fish tissues was found to increase with chlorpyrifos concentrations. Histopathological lesions appeared to be good biomarkers of chlorpyrifos toxicity.

Proposal of Brotulella n. gen. for Monogeneans from the Gills of the Pacific Bearded Brotula Brotula clarkae Hubbs, 1944 (Ophidiiformes: Ophidiidae) Based on Morphological and Molecular Evidence

Based on morphological and molecular data, Brotulella n. gen. is proposed to accommodate the dactylogyrid monogeneans Brotulella laurafernandae n. sp. (type species) and Brotulella luisahelenae n. sp. on the gill filaments of the Pacific bearded brotula Brotula clarkae Hubbs, 1944 (Ophidiiformes: Ophidiidae) from Puerto Pizarro in the Tumbes region (northern Peru). Species of the new genus are distinguished from all other dactylogyrids by the combination of the following features: (1) anchors with a stocking-shaped sheath associated with the distal end of superficial and deep roots, (2) tandem gonads, (3) a vas deferens looping left intestinal caecum, (4) a distally twisted male copulatory organ (MCO) with a delicate membranous accessory piece articulated to the shaft of the MCO, (5) a U-shaped ovary, (6) an almost sigmoid seminal vesicle, and (7) two prostatic reservoirs with thick muscular walls. A 28S ribosomal DNA-based phylogenetic analysis (Maximum likelihood and Bayesian inference) of sequences of two new species of Brotulella n. gen. from the Southeastern Pacific Ocean, along with sequences from closely related genera of the marine Dactylogyridae, supports the establishment of the new genus for the dactylogyrid parasites on the gills of the Pacific bearded brotula.

Occurrence of Multispecies Myxozoan Parasite Infestation by Genus <i>Thelohanellus</i> Kudo, 1933 in a Group of <i>Labeo rohita </i>(Hamilton, 1822)

We describe two Thelohanellus Kudo spp. namely, T. bifurcata (34.94x 8.9μm) on gills and T. caudatus (13x8.9μm) on the caudal and dorsal fins, found infecting Labeo rohita (Hamilton, 1822) fingerlings in a wet lab facility of West Bengal India. The L. rohita fingerlings having spores on fins and gills were segregated and gross pathology was observed for one month. It was done to characterise the morphology of mature spores and to examine the histopathology of infected gills. There were 5-6 white cysts (1-2 mm in length) on the gill filament that were mostly persistent during the one-month study period and infected fish (12.5% prevalent) were listless, anorexic, and emaciated before eventually all of them perished. Spores of 4- 7 mm size in 1-4 numbers were found on caudal and dorsal fins which were visible for two weeks before it gets mature and released, whereas infected fish (20%) were active and underwent normal growth. Additionally, it was discovered that 5% of fingerlings had cysts on their gills and fins. The distal tip of the primary gill lamellae is where the gill epithelium ruptures, releasing the spores. The histological inspection of the infected gill clearly demonstrates infective sporoplasm entry, presporogonic stage, plasmodial maturation, and release of spores. Within a month, it was discovered that 100% of the fingerlings with infected gills died. This is because the plasmodial spores on primary gill filaments damage the structural framework of the gill filament and cause epithelial and vascular cell displacement. Our comparative analysis with previous studies support that orders and families of the parasites are strongly correlated with Thellohanellus Kudo sp: a pathogenic strain of T. bifurcata and a non-pathogenic strain of T. caudatus infecting L. rohita. The spore-forming stages of the gill-damaging T. bifurcata (entry, pre-sporogonic, and sporogonic) are extremely pathogenic to fish fingerlings and cause significant damage to gill tissue

Toxic effects of triclocarban on the histological morphology, physiological and immune response in the gills of the black tiger shrimp Penaeus monodon

Triclocarban (TCC) is a widely used broad-spectrum antimicrobial agent that has become a pollutant threatening the health of aquatic animals. However, the toxic effects of TCC on Penaeus monodon are still lacking. In this study, we exposed P. monodon to 1 μg/L (TCC-1) and 10 μg/L TCC (TCC-10) for 14 days, and the changes of histological morphology, physiological and immune responses in the gills were investigated. The results showed that TCC exposure caused the deformation of the gill vessels and the disordered arrangement of the gill filaments. Oxidative stress biochemical indexes such as H2O2 content, CAT and GPx activity and the relative expression levels of antioxidant-related genes (SOD, GPx and Nrf2) were increased in the TCC-1 and TCC-10 groups; the levels of CAT and HSP70 genes were increased but POD activity was decreased in the TCC-10 group. The relative expression levels of endoplasmic reticulum (ER) stress indexes such as ERP15 and ATF-6 genes were increased in the TCC-10 group, while the level of GRP78 gene was decreased in the TCC-1 and TCC-10 groups. The relative expression levels of apoptosis indexes such as p53 and JNK genes were increased, but CytC and Casp-3 genes were decreased in the TCC-1 and TCC-10 groups. Furthermore, the relative expression levels of detoxification metabolism-related genes (cytP450 and GST) and osmotic regulation-related genes (NKA-α, NKA-β, CA, AQP, CLC and CCP) were increased in the TCC-10 group. The results showed that TCC exposure could affect the physiological homeostasis in the gills of P. monodon, probably via damaging histological morphology, inducing oxidative stress, and disordering ER stress, apoptosis, detoxification and osmotic regulation.

Assessment of Salinomycin's Potential to Treat Microcotyle sebastis in Korean Rockfish (Sebastes schlegelii).

Aquaculture, a crucial sector of the global food industry, faces a myriad of issues due to parasitic invasions. One such parasite, Microcotyle sebastis, which afflicts Korean rockfish in South Korea, has a significant economic impact. The impending danger of resistance to traditional anthelmintics necessitates the exploration of new antiparasitic candidates. Although the efficacy of salinomycin against aquatic parasites such as ciliates and sporozoans is known, its influence on monogeneans has yet to be studied. Therefore, this study investigated the efficacy and safety of salinomycin for the treatment of M. sebastis infections, presenting the first exploration of salinomycin's therapeutic potential against monogeneans. In vitro examinations revealed a minimum effective concentration of salinomycin of 5 mg/kg, which led to necrosis of the haptor upon dislodging from the gill filaments. The one-time oral administration of the drug at concentrations of 5 mg/kg and 10 mg/kg showed a significant dose-dependent reduction in parasite counts, with no apparent behavioral side effects in Korean rockfish. Biochemical analyses monitored the liver, heart, and kidney enzymes, specifically aspartate transaminase (AST), alanine transaminase (ALT), blood urea nitrogen (BUN), and creatine kinase-myocardial band (CK-MB). At both 20 °C and 13 °C, no significant differences were observed in the levels of AST and ALT. However, at 20 °C, alterations in BUN levels were evident on Day 14, a deviation not observed at 13 °C. The CK-MB analysis revealed elevated enzyme levels at both temperatures when compared to the control group, reflecting the similar changes observed in terrestrial animals administered salinomycin. The biochemical data suggest that the oral administration of salinomycin is potentially more favorable at 13 °C than at 20 °C. Although our findings warrant further comprehensive studies, including on the long-term and potential effects on nontarget species and water quality, they also suggest that salinomycin could be considered as an alternative or adjunctive treatment if resistance to the currently used praziquantel against M. sebastis is confirmed.

Anesthetic effect and acute toxicity of Citrus sinensis essential oil in betta

This study investigated the use of Citrus sinensis essential oil (EOCS) in Betta splendens, evaluating toxicity, induction and recovery times to anesthesia, its action on agonist behavior, and on male collective transport. To toxicity, fish were exposed to EOCS at different concentrations during 48 h. The induction test was performed to evaluate sedation, anesthesia, and recovery under different concentrations. To assess agonist behavior, male subjects were maintained with different dosages of EOCS, assessing opercular beat and fin expansion, while exposed to untreated animals. In the mass transport study, males were divided into two treatments with two levels of EOCS, for 6 h, also analyzing gill histometry at the end of transport. The results obtained show that the mean lethal concentration in 48 h of exposure to EOCS was calculated at 49.17 μL·L−1. The shortest anesthesia induction time was at 300 μL·L−1 EOCS. EOCS was able to reduce agonist behavior in males individual. Histometric analyzes revealed a reduction in the height of gill filaments in fish transported with 20 μL·L-1 EOCS. EOCS can be used as a sedative and anesthetic agent for B. splendens.

Some morphological structures and histological features of the gills of Binni (Mesopotamichthys sharpeyi)

This study aimed to describe some morphological and histological aspects of the binni fish's gills. Fifteen adult male binni were pulled alive from the Al-Forat River at about (24 - 28) months, immediately after death. The gills of binni were situated on each side of the head beneath a gill cover; the operculum and the gills were made of finger-like long filaments joined to the cartilaginous gill bar. From each filament, numerous fragile lamellae that resemble leaves protrude. These lamellae comprised tiny capillaries covered in plain squamous epithelial cells. The epithelium acts as a barrier between the fish's blood and the surrounding water. The first, second, third, and fourth pairs of gill arches make up the gills supported by the bone skeleton. Each arch carries a row of gill filaments on its convex side and two rows of gill rakers on its concave side. On the rostral concave border, the filaments in the two extremes of the gill were short, but the length of all the filaments was the same. The first-gill arch's lateral rakers were longer than the medial rakers, while the fourth gill arch's medial rakers were longer than the lateral rakers. The rakers, bony comb-like projections, filled the rostral region of the gill arches at the rostral concave interior side. There was interdigitation between the gill rakers of the neighboring gill arches. Each gill arch included the lateral gill rakers pointing dorsolaterally and medial gill rakers pointing dorsomedially. Filaments originate from the gill arches and are maintained by the cartilage on the caudal convex border of the primary lamellae, from which the secondary lamellae emerge. The simple squamous epithelium lined the secondary lamellae. Keywords: Binni, Gill, Morphology, Histology

A role for dopamine in control of the hypoxic ventilatory response via D2 receptors in the zebrafish gill.

Dopamine is a neurotransmitter involved in oxygen sensing and control of reflex hyperventilation. In aquatic vertebrates, oxygen sensing occurs in the gills via chemoreceptive neuroepithelial cells (NECs), but a mechanism for dopamine in autonomic control of ventilation has not been defined. We used immunohistochemistry and confocal microscopy to map the distribution of tyrosine hydroxylase (TH), an enzyme necessary for dopamine synthesis, in the gills of zebrafish. TH was found in nerve fibers of the gill filaments and respiratory lamellae. We further identified dopamine active transporter (dat) and vesicular monoamine transporter (vmat2) expression in neurons of the gill filaments using transgenic lines. Moreover, TH- and dat-positive nerve fibers innervated NECs. In chemical screening assays, domperidone, a D2 receptor antagonist, increased ventilation frequency in zebrafish larvae in a dose-dependent manner. When larvae were confronted with acute hypoxia, the D2 agonist, quinpirole, abolished the hyperventilatory response. Quantitative polymerase chain reaction confirmed expression of drd2a and drd2b (genes encoding D2 receptors) in the gills, and their relative abundance decreased following acclimation to hypoxia for 48h. We localized D2 receptor immunoreactivity to NECs in the efferent gill filament epithelium, and a novel cell type in the afferent filament epithelium. We provide evidence for the synthesis and storage of dopamine by sensory nerve terminals that innervate NECs. We further suggest that D2 receptors on presynaptic NECs provide a feedback mechanism that attenuates the chemoreceptor response to hypoxia. Our studies suggest that a fundamental, modulatory role for dopamine in oxygen sensing arose early in vertebrate evolution.

Microcotyle justinei n. sp. (Monogenea: Microcotylidae) from the Gills of the Cardinal Fish Apogon imberbis (Teleostei: Apogonidae) off the Algerian Coast of the Western Mediterranean.

A new monogenean Microcotyle justinei n. sp. (Monogenea: Microcotylidae) is described based on specimens found on the gill filaments of the cardinal fish Apogon imberbis (Apogonidae) off the Algerian coast of the Western Mediterranean. Monogeneans were examined, measured and drawn for a comparative morphological study with other species of Microcotyle and characterised molecularly using a partial fragment of the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene. The identification of fish was confirmed by molecular barcoding using the cox1 gene. The new species is distinguished from all other species of the genus by a combination of features, such as the number and size of the clamps, the shape and size of the genital atrium and the number of testes. The molecular analysis of the cox1 gene sequences showed that interspecific differences between Microcotyle justinei n. sp. and published sequences of Microcotyle spp. was greater than 8.8%, strongly suggesting that the new species is distinct from other congeners with sequences available on GenBank. The morphological and molecular analyses support the status of M. justinei as a new species. The present finding extends the list of Microcotyle spp. to 72.

Characteristics of a gill resection – Regeneration model in freshwater laboratory-reared Atlantic salmon (Salmo salar)

Impaired gill function is a critical issue in aquaculture due to factors such as poor water quality, disease, and farm management practices which can greatly compromise gill integrity and ultimately fish growth and survival. Regeneration of damaged gill tissue has been reported for some fish species, however, little is known about Atlantic salmon, which is one of the most economically important salmonid species. This study investigated the regenerative capability of gill tissue following resection in post-smolt Atlantic salmon reared in fresh water. Fish were subjected to two levels of resection severity: 30% or 50% of filament length. Changes in filament length and the cytological and functional capacity of regenerated tissue were studied at several time points post-resection (PR). On average, 38% ± 3.89 of the resected filament length regenerated in both resected groups at week-20 PR. However, absolute regeneration (mm) in 50% resected filaments was greater as compared to the 30% resected filaments. In both groups, regeneration percentage was significantly different among individuals, but was not correlated with initial body size, condition factor, or overall fish growth rate. Immunohistochemistry showed an increase in cell proliferation at the filament tip shortly after resection followed by the appearance of ionocytes, chemoreceptive neuroepithelial cells (NECs), and nerve fibres at week-4 PR. In both resected groups >50% and 90% of NECs were innervated by week-12 and -20 PR, respectively. Gill Na+/K+-ATPase activity in regenerated tissue was not significantly different from that of internal control and non-resected control fish. Our results demonstrate significant regeneration and apparent functional recovery of resected gill filaments in Atlantic salmon which has important implications for recovery from gill damage and developing best practices for maintenance of gill health of farmed salmon.

Histopathological observation and comparative transcriptome analysis reveal immune response mechanisms to Aeromonas dhakensis infection in Macrobrachium rosenbergii

The Macrobrachium rosenbergii industry is threatened by various Aeromonas, resulting in high mortality of adult prawns. However, there are few studies on the immune response of M. rosenbergii infected with Aeromonas dhakensis. In this study, we observed the hepatopancreas and gills histopathologically, performed a comparative transcriptome analysis of the hepatopancreas, and analyzed the candidate gene expression of immune-related genes in the hemolymph, hepatopancreas, and gills of M. rosenbergii that had been infected with A. dhakensis. Histopathology revealed the hepatopancreas was successively inflamed, followed by cellular vacuolation, lumen deformation, and finally tissue erosion; partial and severe inflammation of the gills occurred successively, and eventually the gill tissue atrophy and the gill filaments detached from the gill arch. Transcriptome analysis showed that a total of 77,742 unigenes and 8664 differentially expressed genes (DEGs), and the immune-related DEGs were mainly enriched in lysosome and phagosome pathways. In addition, 4 immune-related candidate genes (RhoA, CASP9, PKC, and DSCIGN) based on KEGG and PPI analysis were monitored at 6, 12, and 24h post injection (hpi) in hepatopancreas, hemolymph and gills. Their spatio-temporal expression results indicated that A. dhakensis have activated the immune system of M. rosenbergii. The present study may provide new information on the complex immune mechanism of M. rosenbergii.

Responses of Two Life Stages of Oreochromis niloticus to Acute Toxicity of Ridoff (Organophosphat Insecticide)

The acute toxicity of Ridoff (Insecticide dichlorvos) on two life stages of Oreochromic niloticus of mean standard length and weight (fingerlings - 4.20 ± 0.60 cm and weight - 7.00±0.20g and juveniles - 12.04 ± 0.20cm and 14.04±0.05g) were investigated under laboratory condition at the concentrations of 0.05, 0.08, 0.10, 0.13 and 0.15ml/l. All the physicochemical parameters (ammonia, dissolved oxygen, pH, temperature and nitrite) under consideration were within acceptable limits. LC50 of Ridoff was calculated at 24, 48, 72 and 96 hours using probit test. Mortality was observed in all treatments of the two life stages exposed from 0.08mg/l at 96 hours. In the first life stage (fingerlings), LC50 was 0.058 within 24 and 96hours and the second life stage (post fingerlings) LC50 was 0.019 within 24 and 96 hours. At a close watch the two life stages exhibited agitated swimming, loss of equilibrum, air gulping, period of quiescence, and turned on flank and swarm in circles and finally died. Accumulation of mucus also was observed on the gill filaments and body surface of the dead fish on exposure to toxicant. Ridoff (Dichlorvos) indeed is harmful to Oreochromis niloticus, which could be a non-target organism in an aquatic ecosystem

Integration of transcriptome and metabolome reveals molecular mechanisms responsive to cold stress in gynogenetic mrigal carp (Cirrhinus mrigala)

Cold stress is a major environmental stimulus affecting the physiological and metabolic activities of farmed fish. In our previous study, we obtained gynogenetic mrigal carp (Cirrhinus mrigala, GMC) with significantly improved cold tolerance compared with the wild individuals. To gain a comprehensive and unbiased understanding of the molecular mechanism of cold tolerance in GMC, transcriptomes and metabolomics of liver tissues were comparatively analysed. After cold treatment at 14 °C for 48 h, histology analysis revealed that the hepatocyte nucleus was shifted, blurred and disintegrated; the terminus of the gill filament was enlarged; and the number of mucus cells and their secretions were increased. A total of 4844 differentially expressed genes (DEGs), including 2420 upregulated and 2424 downregulated genes, were identified. The upregulated KEGG pathways were mainly involved in oxidative phosphorylation, the proteasome and the citrate cycle, while the downregulated pathways were involved in staphylococcus aureus infection, systemic lupus erythematosus and leishmaniasis. A total of 160 differential metabolites (DMs) were identified, including 39 increased and 121 reduced metabolites. The KEGG pathway analysis revealed significant enrichment in retinol metabolism and folate biosynthesis. KEGG function analysis showed that the DEGs and DMs were both highly enriched in purine metabolism, neuroactive ligand–receptor interaction and galactose metabolism. Correlation analysis between DEGs and DMs revealed that multiple pathways were active and strongly related to immunity and disease, metabolism and growth under cold stress. This study can facilitate our understanding of the molecular mechanisms of fish response to cold stress and provide insight into the application of gynogenesis in farmed fish.

Multi-Effects of Acute Salinity Stress on Osmoregulation, Physiological Metabolism, Antioxidant Capacity, Immunity, and Apoptosis in Macrobrachium rosenbergii.

Salinity stress can trigger a series of physiological changes. However, the mechanism underlying the response to acute salinity stress in Macrobrachium rosenbergii remains poorly understood. In this study, osmoregulation, physiological metabolism, antioxidant capacity, and apoptosis were examined over 96 h of acute salinity stress. Hemolymph osmolality increased with increasing salinity. After 48 h of salinity exposure, the glucose, triglycerides, total protein, and total cholesterol contents in two salinity stress groups (13 and 26‱ salinity) were significantly lower than those in the 0‱ salinity group. The highest levels of these parameters were detected at 6 h; however, superoxide dismutase (SOD), total antioxidant capacity (T-AOC), and malondialdehyde (MDA) were the lowest at 96 h in the 13‱ salinity group. The activity of immunity-related enzyme alkaline phosphatase (AKP) showed a decreasing trend with increasing salinity and remained at a low level in the 26‱ salinity group throughout the experiment. No significant differences were observed in aspartate aminotransferase (AST), alanine aminotransferase (ALT), or lysozyme (LZM) among the three treatments at 96 h. After 96 h of salinity treatments, the gill filament diameter significantly decreased, and a more pronounced terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL)-positive signal was detected in the 13‱ and 26‱ groups compared to that in the 0‱ group. Expression levels of apoptosis-related genes, including Cysteine-aspartic acid protease 3 (Caspase 3), Cysteine-aspartic acid protease 8 (Caspase 8), Cytochrome c (Cyt-c), tumor suppressor gene (P53), Nuclear factor kappa-B (NF-κB), and B cell lymphoma 2 ovarian killer (Bok) were significantly higher in the 26‱ salinity group than in the other groups at 24 h, but lower than those in the 0‱ salinity group at 96 h. Cyt-c and P53 levels exhibited a significantly positive relationship with MDA, AST, and LZM activity during salinity stress. In the 13‱ salinity group, Bok expression was significantly correlated with SOD, T-AOC, AKP, acid phosphatase, and LZM activity, whereas in the 26‱ group, the AST content was positively correlated with Caspase 8, Cyt-c, and P53 expression. A significant negative relationship was observed between Caspase 3 expression and catalase (CAT) activity. These findings provide insight into the mechanisms underlying the response to acute salinity stress and will contribute to improving M. rosenbergii aquaculture and management practices.

POTASSIUM CHLORIDE ADMINISTERED VIA FOUR ROUTES FOR EUTHANASIA OF ANESTHETIZED GOLDFISH (CARASSIUS AURATUS).

Immersion overdose in tricaine methanesulfonate (MS-222) is ineffective for euthanasia of adult goldfish (Carassius auratus), and investigation of alternative techniques is warranted. This study evaluated potassium chloride (KCl) administered via four routes for goldfish euthanasia. Thirty clinically healthy adult goldfish (17.2-41.9 g) were anesthetized via immersion in buffered MS-222 (300 mg/L) for 5 min beyond anesthetic induction. Fish were then randomly administered one of five treatments: KCl (333 mg/mL, 4.5 mmol/mL, 4.5 mEq/mL) via intracardiac injection at 10 mEq/kg (IC), intracoelomic injection (ICe) or bilateral topical delivery over the gill filaments (T) at 100 mEq/kg, or 90-min immersion at 4,500 mEq/L (saturated solution) dissolved in induction solution (water [W]), or no treatment (control [C]). Following treatment, all fish were moved to anesthetic-free freshwater. Serial heart rates were collected via Doppler device until sound cessation or recovery. Median (range) time to perform treatments was 5.3 (1.2-8.0) min, 3 (3-3) s, and 10 (10-10) s in IC, ICe, and T, respectively. Doppler cessation occurred in 6/6, 6/6, 6/6, 6/6, and 0/6 fish in median (range) times of 3 (0-210), 18 (10-45), 118 (90-390), and 150 (60-240) min in IC, ICe, T, W, and C, respectively; this was significantly different between groups (P = 0.002). Following or during treatment, 1/6, 2/6, 6/6, and 4/6 fish in IC, ICe, T, and W, respectively, exhibited intermittent transient jerking movements. Median (range) time to recovery in C was 5.5 (3.5-6.5) min. All administration routes were effective, but time to Doppler cessation varied and transient movements were noted. Preliminary follow-up research determined that return to MS-222 induction solution following ICe KCl treatment abolished movements. ICe KCl at 100 mEq/kg was technically simple and resulted in Doppler sound cessation in a clinically relevant time frame in 6/6 anesthetized goldfish. Return to MS-222 immersion following KCl administration is recommended.

Physiological responses to acute warming at the agitation temperature in a temperate shark.

Thermal tolerance and associated mechanisms are often tested via the critical thermal maximum (CTMax). The agitation temperature is a recently described thermal limit in fishes that has received little mechanistic evaluation. The present study used a temperate elasmobranch fish to test the hypothesis that this thermal tolerance trait is partially set by the onset of declining cardiorespiratory performance and the cellular stress response. Pacific spiny dogfish (Squalus suckleyi) were screened for cardiorespiratory and whole-organism thermal limits to test for associations between thermal performance and tolerance. Then, biochemical markers of secondary stress, aerobic and anaerobic enzyme activities, and molecular markers of cellular stress were determined for various tissues at the agitation temperature, and secondary stress markers were determined at CTMax. In dogfish, the agitation temperature was characterised by increased turning activity within experimental chambers and was equal to the temperature where dogfish exhibited maximum heart rate. Citrate synthase activity increased at the agitation temperature in white muscle relative to unmanipulated dogfish. Further, lactate dehydrogenase activity and accumulated lactate in the plasma and muscle were not affected by acute warming. Cellular stress was apparent in hypothalamus, gill filament, and ventricle, denoted by elevated transcript abundance of hsp70, but not hif1α. Conversely, CTMax was characterised by metabolic acidosis driven by anaerobic lactate production, signifying an increased reliance on anaerobic metabolism between the agitation temperature and CTMax. Together, these data provide partial support for our hypothesis, in that cellular stress, but not declining thermal performance, occurred at the agitation temperature.

Live Freshwater Parasite, Salmincola californiensis (Copepoda: Lernaeopodidae), on the Gills of an Ocean-Migrating Steelhead Trout (Oncorhynchus mykiss) and Discussion on the Origin and Survival of the Parasite at Sea.

Salmincola californiensis is a parasitic copepod of freshwater salmonids in the North Pacific rim countries. Sixteen adult females of the species were found alive on the gills of an ocean-age 4, maturing steelhead trout, Oncorhynchus mykiss, caught in offshore waters (50°30'N, 179°30'W) of the North Pacific Ocean in July 1997. This is the first evidence of live individuals of S. californiensis from ocean-migrating salmonids. When found, copepods were attached to the distal ends of gill filaments, and their bodies were observed to be slowly moving in Petri dishes with seawater. Ocean-migrating steelhead trout comprise individuals originating from western Kamchatka (Russia) and western North America. Based on the date and catch location of the infected fish, it is inferred that it originated from western North America, where it acquired S. californiensis infection in fresh water. As this fish spent about 4 years in the ocean, the copepods likely survived the same period at sea. However, if the fish was a kelt, the survival period of the copepods in the ocean may be shorter than four years. To confirm identification of the copepods, adult females of S. californiensis are briefly described using the specimens collected from the fish.

Гістологічна структура зябер карася сріблястого річки Самара Дніпропетровської області

Purpose. To study the histological structure of the gill apparatus of the Prussian carp (Сarassius gibelio) from the point of view of the presence of pathologies with differentiation according to areas with different levels of anthropogenic pressure of the Samara River in the Dnipropetrovsk region. Methodology. The generalized sutdy results were obtained during 2019–2021. The study was conducted within three sampling sites with different types of human impact. The object of the study were age-6 Prussian carps of both sexes. Histological studies were performed in accordance with generally accepted methods. Gills were obtained by anatomical dissection from freshly caught fish. Photographs of histological specimens were taken using a digital camera “SciencelabT5005.17M” and an microscope “Ulab XY-B2TLED”. Measurements of the samples were performed using the “ScienceLabView7”. Statistical processing of the obtained data was carried out in MS Excel 2010. Findings. The conducted histological studies showed that pathological changes in the structure of the gill apparatus were observed in age-6 Prussian carps in all sampling sites of the Samara River. In particular, such pathologies as curvature of lamellae terminals, accumulation of mucus and hyperplasia of the epithelium of gill filaments, fusion of lamellae were found. Single cases of necrosis and twisting of lamellae threads were recorded. Most often, histopathological changes in the gill apparatus were observed in fish caught near the city Novomoskovsk. Organometric studies showed that there was no significant difference between the length and width of gill arches of Prussian carp from different sampling sites. The length and width of the lamellae of the gill apparatus was the largest in individuals sampled near the city Novomoskovsk, which may indicate a greater need for oxygen for the fish body in the specified area. The length of the respiratory lamellae was the lowest in Prissian carp sampled near the village of Novoselivka. The lowest width of the lamellae were found in fish from the village of Khashcheve. The obtained data give reason to conclude that the conditions of existence for Prussian carp are most favorable near the village of Novoselivka. Originality. For the first time the histological structure and pathology of the gills aparatus of Prussian carp in the conditions of the Samara River in the Dnipropetrovsk region was studied. Practical value. The obtained data on the histological changes of the gill apparatus of fish can be used to identify adaptive reactions, will allow assessing the impact of anthropogenic factors on hydrobionts, and will also provide the opportunity to monitor the morpho-physiological state of a specific population under the impact of pollutants of various origins. Key words: Prussian carp, gill apparatus, morphological and histological indicators.

Morphological, histological and phylogenetic identification of three species of Myxobolus (Myxosporea: Myxobolidae) infecting different carp lineages in China.

The global cultivation of common carp Cyprinus carpio is developed primarily for either food or recreational purposes and myxosporean infections causing significant economic losses are regularly troublesome for carp farmers. However, most myxosporean species are poorly characterized, making it difficult to correctly elaborate on their parasitism mode and determine pathogenicity. Here, based on an integrative approach, we supplemented fundamental information on three Myxobolus species infecting different carp lineages in China. Myxobolus elliptoides Wu et Chen, 1987; Nine new species of Myxosporida from freshwater fish of Wuhu lake, Hubei, China. Acta Hydrobiologica Sinica, 11, 161 formed yellowish pseudocysts in the anal fin of common carp and were observed with ellipsoidal myxospores and two polar capsules of distinct dimensions. The dermal tissue within the fin ray was regarded as the type site for growing plasmodia of M. elliptoides. Myxobolus basilamellaris Lom et Molnar, 1983; Myxobolus basilamellaris sp. n. (myxozoa, Myxosporea), a parasite of the gills of common carp (Cyprinus carpio L.). Folia Parasitologica, 30, 1 aggregated ellipsoidal myxospores at the base of the gill filaments of koi carp juvenile, causing significant tissue damage. Myxobolus artus Achmerov, 1960, enwrapped in diffuse pseudocysts, was observed throughout the trunk muscle of mirror carp. Its oblate myxospores were asymmetrical and contained two unequal-sized polar capsules. According to the localization of large plasmodia, inter- and intramuscular preferences were displayed by M. artus. For precise species identification, we provided the SSU rDNA sequences for each species of Myxobolus. Among them, M. elliptoides was molecularly characterized for the first time, showing the highest 94.21% identity to Thelohanellus sinensis (KY469292). Phylogenetically, the affinity of both M. artus and M. basilamellaris to their conspecific species derived from different carp lineages was highly supported. Moreover, the intermixed cluster of Myxobolus species, including M. elliptoides, with those of Thelohanellus species provided evidence for querying the monophyletic evolution history of these taxa. This work updates the appreciation of the concerned species and enhances our understanding of the parasite fauna of common carp.

Morphological, histopathological, ultrastructural and phylogenetic analysis of Henneguya archosargus n. sp. (Cnidaria, Myxosporea) infecting the sparid fish Archosargus probatocephalus in Brazilian waters

The introduction of new fish species to the aquaculture industry is essential to halt the progressive decline of natural fish stocks. The sheepshead Archosargus probatocephalus is a commercially valuable sparid fish with potential for breeding in captivity, but with limited information regarding parasitic infections that could pose a significant threat for its sustainable production. Thus, the present study aimed to study the myxozoan diversity infecting A. probatocephalus. A novel Henneguya sp. was detected forming plasmodia in the gill lamellae of specimens inhabiting the Brazilian coast, and is characterized based on morphological, histopathological, ultrastructural, molecular, and phylogenetic data. Myxospore total length was 21.3 ± 0.8 μm, with myxospore body 10.0 ± 0.5 μm long, 6.2 ± 0.3 μm wide, and 4.8 ± 0.5 μm thick. Caudal appendages were 10.3 ± 0.5 μm long and did not present any type of coating. Two pyriform polar capsules, 3.4 ± 0.3 μm long and 1.5 ± 0.2 μm wide, each containing an isofilar polar tubule with 4–5 coils. Histopathological analyses showed large intralamellar polysporic plasmodia associated with vascular congestion of the gill filament and gill lamellae, as well as epithelial hyperplasia causing partial or total fusion of gill lamellae. Maximum likelihood and Baysesian inference SSU rDNA-based phylogenetic analyses showed the novel sequence grouped within the marine clade of Henneguya spp. that mostly parasitize fishes belonging to Eupercaria incertae sedis.