Ciguatera is a foodborne disease caused by ciguatoxins (CTXs), produced by dinoflagellates (genera Gambierdiscus and Fukuyoa), which bioaccumulate in fish through the food web, causing poisoning in humans. Currently, the physiological mechanisms of the species with the highest amount of toxins in their adult stage of life that are capable of causing these poisonings are poorly understood. Dusky grouper (Epinephelus marginatus) is a relevant fishing species and is part of the CTX food chain in the Canary Islands. This study developed an experimental model of dietary exposure featuring adult dusky groupers with two diets of tissue naturally contaminated with CTXs (amberjack and moray eel flesh) with two different potential toxicities; both groups were studied at different stages of exposure (4, 6, 10, 12, and 18 weeks). The results showed that this species did not show changes in its behavior due to the provided feeding, but the changes were recorded in biochemical parameters (mainly lipid and hepatic metabolism) that may respond to liver damage and alterations in the homeostasis of the fish; more research is needed to understand histopathological and cytotoxic changes.

Ceratomyxa marginati n. sp. and C. aenei n. sp. are two new coelozoic myxosporean species infecting the gallbladder of the dusky grouper (Epinephelus marginatus) and the white grouper (Epinephelus aeneus), respectively. These two ceratomyxids were described using morphological characteristics and molecular analysis of the SSU rDNA. Ceratomyxa marginati n. sp. exhibits disporic plasmodia measuring 12-14 µm long and 11-12 µm wide and mature myxospores which are slightly crescent-shaped, measuring 6.0 ± 0.3 (5.6-6.8) µm in length and 12.9 ± 0.9 (11.5-14.0) μm in thickness. Plasmodia of C. aenei n. sp. were disporic and ellipsoidal and measured 28-32 μm in length and 19-22 μm in width. Mature myxospores of C. aenei n. sp. were elongated with unequal shell valves and measured 7.4 ± 0.6 (6.9-9.0) μm in length and 26.9 ± 2.4 (23.2-30.0) μm in thickness. Based on the SSU rDNA sequences, Ceratomyxa marginati n. sp. and C. aenei n. sp. are distinct from all other Ceratomyxa sequences available in GenBank. Phylogenetic analysis reveals that the two new species branched together within a clade with other Ceratomyxa species from different host families and different geographical localities with maximum support (100%).

The current study has designed to investigate the morphological characteristics of the gills of the dusky grouper (Epinephelus marginatus) by gross morphology, scanning electron microscopy, and morphometric analysis. The study was carried out on 10 fresh fish. The fish has four-gill arches on each side. The lengths of the first-, second-, third-, and fourth-gill arches were 5.27, 4.2, 3.2, and 2.8 cm. The gill arches carried a longitudinal band, the bases of the gill filaments, and gill rakers that varied from rectangular to circular shapes. Each gill arch had two main lateral and medial rows and two accessory rows of gill rakers in an alternative manner with each other. The dusky grouper fish had long rakers, whereas the longest one was the lateral rakers of the first arch, which were 467 and 1271.9 μm in width and length. Three types of spines appeared on the gill arches and rakers. The long spine had detected on the apex of the short rakers. Each gill arch carried on its ventral side two rows of gill filaments. The long filaments were at the middle of the gill arch, while the short ones were at the rostral end of the gill arch. The study has demonstrated the taste buds, mucous, chloride cells, and significant features of the epithelial covering of the gill arches and rakers. The morphology of the gills of the dusky grouper indicates the adaptation to their marine environment.

This study aimed to investigate parasite fauna of E. marginatus from the central Mediterranean Sea between Messina and Syracuse. In the present survey; parasite fauna of dusky grouper was investigated for two main reasons: the economic value of this species and the current lack of studies regarding the capture area. Seventy dusky groupers were caught from May 2018 to February 2020. Forty-seven out of the 70 specimens (67.2%) were infected with one or more parasite species. The most abundant species was Prosorhynchus caudovatus (42.9%), followed by Podocotyle temensis (28.6%), Didymodiclinus sp. (18.6%), Philometra jordanoi (5.7%), Anisakis Type II larvae (5.7%). Higher prevalence of infection of P. jordanoi and Contracaecum sp. was found in warm months (March to September), while P. caudovatus and P. temensis were mostly found during cold months. Weight and total length of E. marginatus were positively correlated with the parasitic load of P. jordanoi and Didymodiclinus sp. The different prevalence of parasite infection found between warm and cold months is probably related to the diet of the dusky grouper; which is characterized by mollusks that are intermediate hosts for parasite species found. None of the parasites found in the present survey is responsible for zoonosis.

Trypanorhyncha are cestodes commonly infecting marine fish. Numerous studies have detailed the biology of Trypanorhyncha species, but information on the pathological changes produced by these parasites is limited. Dusky groupers are keystone species necessary for the preservation of several marine ecosystems. Considering their vulnerable state of conservation and the efforts being made to culture them, identification of the effects caused by Trypanorhyncha is vital. Here, we aimed to determine the prevalence and pathological changes produced by Trypanorhyncha in dusky groupers from the Canary Islands. The prevalence of trypanorhynch plerocerci was 96%. Grossly, in the abdominal cavity, there were numerous larvae-filled cysts and nodules. These were embedded in abundant fibrosis, producing visceral adhesions. Histologically, affecting the peritoneum, stomach, and intestine there were numerous degenerated encysted plerocerci and extensive deposition of mature connective tissue. These findings indicate that Trypanorhyncha is highly prevalent in adult dusky groupers from the Canary Islands, producing a progressive and chronic response. Furthermore, fish immune system appears to attempt to eliminate the parasites through fibrous encapsulation. Nonetheless, extensive fibrosis may have a detrimental impact on fish health when adjacent cells or tissues are compressed and their functions impaired.

Ceratomyxa mennani n. sp. is a new coelozoic Ceratomyxa species found in the gallbladder of Epinephelus marginatus from the Gulf of Tunis, Tunisia. Mature plasmodia were disporic, ovoid in shape measuring 9-12 μm in width and 11-14 μm in length. Mature myxospores were slightly crescent-shaped with almost straight posterior margin, measuring 5.8 ± 0.2 (5.4-6.1) μm in length and 12.7 ± 0.3 (11.9-13.0) μm in thickness. The two valves were unequal with rounded ends. Polar capsules were spherical, equal in size with 2.1 ± 0.2 (1.9-2.6) μm in diameter. The binucleated sporoplasm filled the entire cavity of the myxospore. Molecular analysis of SSU rDNA sequences indicated that C. mennani n. sp. was distinct from all other Ceratomyxa sequences in GenBank. Phylogenetic analysis revealed that C. mennani n. sp. clustered with Ceratomyxa species infecting Epinephelinae fishes. Seasonal prevalence of infection over one year was significantly higher in winter and the lowest in autumn. This is the third report of Ceratomyxa species infecting the gallbladder of Epinephelus marginatus from Tunisia and the first study to include molecular data.

This is the first description of a betanodavirus infection in the dusky grouper Epinephelus marginatus within the marine protected areas (MPAs) of the Balearic Islands. Histopathology techniques were employed to describe neurological lesions in infected fish. Abnormal swimming, mortality, and neurological lesions were detected in all analysed grouper individuals. Virus particles were observed by means of transmission electron microscopy. Reverse transcription of RNA1 and RNA2 followed by cDNA amplification and sequencing allowed viral classification. Phylogenetic analysis showed the isolates from wild E. marginatus of the Balearic Islands MPAs to be closely related to Dicentrarchus labrax and Mullus barbatus strains from Cyprus and Italy. Although vertical transmission from infected spawners has been described as the major route for nodavirus infection, we point out in this work that horizontal transmission among sub-clinical fishes after migration or commercial import for aquaculture production could play a major role in the spreading of the disease in MPAs.

Development, ontogeny of the digestive system and the fatty-acid (FA) profile, were analysed during development of Epinephelus marginatus. Larvae were analysed 7 and 17 days post fertilization (dpf) to evaluate fatty-acid profile and morphological variables, respectively. Epinephelus marginatus larvae have relatively slow development of digestive structures, but were able to capture, ingest and digest prey by 5 dpf. Eggs were composed of high percentages of polyunsaturated fatty acids (PUFA) in phospholipids. The percentage of n3 PUFAs was higher than n6, especially docosahexaenoic acid (DHA), which exhibited higher levels compared with other marine species during the first 3 days of development, both in terms of phospholipids and triglycerides. The larvae present a high content of docosahexaenoic acid-eicosapentaenoic acid (DHA-EPA) and, during this phase, live food of small size was required (copepods or SS-strain Brachionus rotundiformes), enriched with DHA-EPA. These results may guide future studies on the contribution of FAs required during this stage of the life cycle of E. marginatus, to advance knowledge of the use of these FAs throughout ontogeny and contribute to the culture of this species commercial production or restocking.

The effects of halogen-light-irradiated and non-irradiated PAHs on the grouper Epinephelus marginatus were assessed through biomarkers including morphometric parameters, liver histopathology, biliary PAH concentration, genetic alterations, and enzyme activity modulation. E. marginatus juveniles were divided into three groups: control (C), non-irradiated PAHs (PAHs1), and irradiated PAHs (PAHs2). Test groups were exposed for 14 days to a 0.5 ppm PAH solution in the semi-static system. After this period, fish were anesthetized with benzocaine (2%) and peripheric blood was collected by caudal puncture. Blood smears were prepared and stained with propidium iodide. Fish livers were collected, fixed in McDowell\'s solution, embedded in paraplast, thin-sectioned, and stained with hematoxylin-eosin (H&E). For biochemical analyses including superoxide dismutase, catalase, and glutathione S-transferase activities, fish livers were collected and preserved in liquid nitrogen. Water samples were analyzed using gas chromatography-mass spectrometry (GC-MS) and bile synchronous fluorescence spectroscopy. Fish in the PAHs2 group had micronuclei (MN) in blood cells, as well as significant differences in nuclear morphology (NMA). Significant morphological alterations were observed in the livers from fish exposed to PAHs as well as inhibition of the catalase activity. Our results show that irradiation altered the bioavailability of PAHs, especially benzanthracene, which has great impact in aquatic ecosystems. Among the consequences of physical and chemical changes to PAHs, we observed a significant increase in NMA and MN incidence in E. marginatus erythrocytes, indicating the potential initiation of mutagenic and carcinogenic processes.