A novel use of social media to evaluate the occurrence of skin lesions affecting wild dusky grouper,Epinephelus marginatus (Lowe, 1834), in Libyan coastal waters

The social media network Facebook^? was used to gather information on the occurrence and geographical distribution of dusky grouper dermatitis, a skin lesion affecting the dusky grouper, Epinephelus marginatus. Dusky grouper are common targets for spear fishermen in the Mediterranean and by monitoring spearfishing activity in Libyan waters, it was possible to document skin lesions from their entries on Facebook. Thirty‐two Facebook accounts and 8 Facebook groups posting from 23 Libyan coastal cities provided a retrospective observational data set comprising a total of 382 images of dusky grouper caught by spearfishing between December 2011 and December 2015. Skin lesions were observable on 57/362 fish, for which images were of sufficient quality for analysis, giving a minimal prevalence for lesions of 15.75%. Only dusky grouper exceeding an estimated 40 cm total length exhibited lesions. The ability to collect useful data about the occurrence and geographical distribution of pathological conditions affecting wild fish using social media networks demonstrates their potential utility as a tool to support epidemiological studies and monitor the health of populations of aquatic animals. To our knowledge, this represents the first time that such an approach has been applied for assessing health in a wild population of fish.     

Infection and pathology in Queensland grouper,Epinephelus lanceolatus, (Bloch), caused by exposure to Streptococcus agalactiae via different routes

Since 2007, 96 wild Queensland groupers, Epinephelus lanceolatus, (Bloch), have been found dead in NE Australia. In some cases, Streptococcus agalactiae (Group B Streptococcus, GBS) was isolated. At present, a GBS isolate from a wild grouper case was employed in experimental challenge trials in hatchery‐reared Queensland grouper by different routes of exposure. Injection resulted in rapid development of clinical signs including bilateral exophthalmia, hyperaemic skin or fins and abnormal swimming. Death occurred in, and GBS was re‐isolated from, 98% fish injected and was detected by PCR in brain, head kidney and spleen from all fish, regardless of challenge dose. Challenge by immersion resulted in lower morbidity with a clear dose response. Whilst infection was established via oral challenge by admixture with feed, no mortality occurred. Histology showed pathology consistent with GBS infection in organs examined from all injected fish, from fish challenged with medium and high doses by immersion, and from high‐dose oral challenge. These experimental challenges demonstrated that GBS isolated from wild Queensland grouper reproduced disease in experimentally challenged fish and resulted in pathology that was consistent with that seen in wild Queensland grouper infected with S. agalactiae.     

Can barber goby Elacatinus figaro control Neobedenia melleni infections on dusky grouper Epinephelus marginatus?

The interaction between cultured barber goby Elacatinus figaro (cleaner fish) and the dusky grouper Epinephelus marginatus, and the efficiency of the cleaner fish in removing ectoparasites were evaluated. When the interaction between these two species was observed, cleaner fish showed a preference for the largest groupers. In a second trial, treatments: TWC (Control) – two groupers without a cleaner fish, T1C – two groupers with one cleaner fish, T3C – two groupers with three cleaner fish and T6C – two groupers with six cleaner fish were tested in four replicates. After 8 days, monogeneans were removed and identified as Neobenedenia melleni. The highest mean abundance of parasites was found on the groupers in the TWC group (37 parasites per host) and the lowest on those in the T6C group (4.1 parasites per host). By increasing the number of cleaner fish, a higher cleaning efficiency was obtained, as observed in T6C, where almost 90% of the parasites were removed. Possibly, this removal would have been complete if the number of cleaners had not been reduced in the treatments due to the mortalities observed. This study demonstrates the possibility of using gobies to remove monogeneans and in improving grouper health.     

Induced sex reversal of dusky grouper, Epinephelus marginatus (Lowe)

The present paper describes sex reversal in dusky grouper, Epinephelus marginatus (Lowe). Over a 4-month feeding period, 5 mg kg^?1 of the male hormone 17-α methyltestosterone was combined with frozen fish as feed. After 2.5 months of treatment, the gonads of all six fish treated in the experiment presented tissues with oocytes and spermatogonia, and after 3 months, there were only a few remnants of oogonial tissue left in the gonads. All fish in the experiment gave sperm of varying quality after 4 months of the trial. The quantity of the hormone administered to the fish over these 4 months was 89-113 mg kg^?1. Spermatozoid motility varied from 95% to 40%, and lasted for about one hour in sea water. Using the combined sperm of three males, fertilization ranged from 50% to 95%. Fertilized eggs and hatched larvae continued to develop normally.     

Prevalence,intensity and histopathology of Zeylanicobdella arugamensis infestation on groupers reared on different aquaculture systems

The marine leech, Zeylanicobdella arugamensis, is a major threat to aquaculture in grouper-producing countries including Indonesia. This study aimed at investigating prevalence, intensity and histopathology of the ectoparasite in humpback and hybrid groupers cultured in different rearing systems. A total of 260 groupers (60 humpback groupers and 200 hybrid groupers) were used for samples. The marine leech was observed on skin, fins, gills and mouth, followed by histopathological assay on the skin tissue. The results showed that prevalence of the leech in both groupers was higher when they were cultured in the floating net cages compared with the hatchery, p < .05. Furthermore, humpback grouper had a higher prevalence than hybrid grouper when they were cultured in a similar system, p < .05. Meanwhile, there was no significant difference in intensity between the two groupers, p > .05. Within the hybrid groupers, the highest prevalence was obtained from hybrid groupers reared in the earthen ponds. Histopathological studies showed that the infected groupers exhibited inflammation, congestion and erosion of the epidermis layer. Hybrid grouper had more severe histopathological lesions in the skin tissues. These results suggested that species and type of aquaculture system had significantly determined the prevalence, intensity and severity of lesion in Z. arugamensis infestation.     

Pathology of red spot disease in sea mullet, Mugil cephalus L., from eastern Australia

Abstract. Lesions present on sea mullet, Mugil cephalus L., during the first 3 weeks of red spot disease outbreaks are described. Necrotizing dermatitis is a severe, locally extensive, granulomatous lesion associated with invasion of dermis and underlying skeletal muscle by numerous, non-septate, fungal hyphae 12–18 μm in diameter. Erythematous dermatitis is a mild to severe, focal, chronic active dermatitis without fungal involvement. Lesions intermediate between these two forms, with small to moderate numbers of fungal hyphae in dermis and skeletal muscle also occur. Findings indicate that erythematous dermatitis lesions and intermediate-type lesions subsequently resolve, while necrotizing dermatitis lesions consistently develop into dermal ulcers, with associated severe necrotizing granulomatous myositis. Generally, dermal ulcers occur significantly more often on posterior and dorsal areas of the body surface than on anterior and ventral areas. Lesions heal by combinations of epidermal and dermal repair, fibroplasia, destruction of fungi, removal of necrotic skeletal muscle and regeneration of myofibres. Atrophy of exocrine pancreas occurs in both diseased and clinically normal fish, but is generally more severe in diseased fish. The possible roles of suspected disease determinants, including falls in dissolved oxygen concentrations to sub-lethal levels prior to red spot disease outbreaks, are discussed.     

Ecological and histopathological aspects of Didymodiclinus sp. (Trematoda: Didymozoidae) parasite of the dusky grouper,Epinephelus marginatus (Osteichthyes: Serranidae), from the western Mediterranean Sea

The dusky grouper Epinephelus marginatus (Lowe) is an ecologically and commercially important fish species of the Atlantic and Mediterranean coastal rocky habitats. Despite records of didymozoid infections in several grouper species, the identification and pathogenesis of these parasites in E. marginatus are lacking. The aim of this study is to characterize the didymozoids of E. marginatus, particularly their mechanisms of infection and histopathological features. Dusky groupers (n = 205) were caught off Majorca Island (western Mediterranean Sea) and examined for parasites. Of the fish sampled, 45% were infected with white and yellow didymozoid capsules and brown nodules, found on the gills and pseudobranchs. Parasite abundance had a strong positive relationship with the fish length; only fish larger than 20 cm were infected, suggesting infection via consumption of an intermediate host, for which E. marginatus size was a limiting factor. The capsules contained two convoluted viable adult trematodes, identified as Didymodiclinus sp., in close contact with host capillary vessels, with no evidence of the tissue inflammatory response. Conversely, nodules containing degraded parasites were surrounded by an intense inflammatory infiltrate. The findings suggest that Didymodiclinus sp. have the potential to evade the host's immune system by inhibiting the inflammatory response.     

Natural outbreak of Streptococcus agalactiae (GBS) infection in wild giant Queensland grouper, Epinephelus lanceolatus (Bloch), and other wild fish in northern Queensland, Australia

Ninety‐three giant Queensland grouper, Epinephelus lanceolatus (Bloch), were found dead in Queensland, Australia, from 2007 to 2011. Most dead fish occurred in northern Queensland, with a peak of mortalities in Cairns in June 2008. In 2009, sick wild fish including giant sea catfish, Arius thalassinus (Rüppell), and javelin grunter, Pomadasys kaakan (Cuvier), also occurred in Cairns. In 2009 and 2010, two disease epizootics involving wild stingrays occurred at Sea World marine aquarium. Necropsy, histopathology, bacteriology and PCR determined that the cause of deaths of 12 giant Queensland grouper, three wild fish, six estuary rays, Dasyatis fluviorum (Ogilby), one mangrove whipray, Himantura granulata (Macleay), and one eastern shovelnose ray, Aptychotrema rostrata (Shaw), was Streptococcus agalactiae septicaemia. Biochemical testing of 34 S. agalactiae isolates from giant Queensland grouper, wild fish and stingrays showed all had identical biochemical profiles. The 16S rRNA gene sequences of isolates confirmed all isolates were S. agalactiae; genotyping of selected S. agalactiae isolates showed the isolates from giant Queensland grouper were serotype Ib, whereas isolates from wild fish and stingrays closely resembled serotype II. This is the first report of S. agalactiae from wild giant Queensland grouper and other wild tropical fish and stingray species in Queensland, Australia.     

Parasites of cultured and wild brown‐marbled grouper Epinephelus fuscoguttatus (Forsskål, 1775) in Lampung Bay,Indonesia

A total of 210 Epinephelus fuscoguttatus (brown‐marbled grouper) was examined for parasites. During three consecutive seasons (two rainy and one dry season from 2002 to 2004), 35 specimens each taken from floating net cages of the National Sea Farming Development Centre (Balai Budidaya Laut) and from wild catches in Lampung Bay, South Sumatra, Indonesia were studied. Twenty‐five (cultured grouper) and 30 (wild grouper) parasite species/taxa were identified, with an infracommunity ranging from one to nine (cultured) and three to 14 parasite species (wild), demonstrating a species‐rich parasite fauna even in the cultured fish. Protozoans (1 species), microsporeans (1), myxozoans (1), digeneans (8), monogeneans (5), cestodes (3), nematodes (8), acanthocephalans (2) and crustaceans (6) were found. The most abundant parasites were the monogeneans Pseudorhabdosynochus epinepheli and Pseudorhabdosynochus lantauensis for both, cultured and wild grouper during all seasons. For the cultured fish, the prevalence of monoxenous ectoparasites (e.g. P. epinepheli, P. lantauensis, Capsalidae gen. et sp. indet., Benedenia epinepheli) was in most cases higher than that of heteroxenous endoparasites. This contrasts the wild grouper, where heteroxenous parasites such as Allopodocotyle epinepheli and Raphidascaris sp. occurred at a similar prevalence compared with the fairly abundant Pseudorhabdosynochus spp. No seasonality of infestation was observed for both cultured and wild fish. The high levels of infestation of potentially pathogenic monogeneans throughout the year could result in significant parasite outbreaks at the locality studied.     

Transport of juvenile dusky grouper Epinephelus marginatus under different packing densities: Metabolic and haematological responses

The aim of this study was to investigate a suitable packing density for the transport of juvenile dusky grouper, Epinephelus marginatus, based on the evaluation of stress responses in blood. After acclimation, fish were placed in plastic bags and transported for 8 hr on paved road at densities of 28, 45 and 64 g/L. Water quality was monitored before and after transport. Blood was collected before, upon arrival (0 hr), after 2 and 24 hr of transport. Plasma cortisol, blood glucose, partial pressures of O₂ (pO₂) and CO₂ (pCO₂), blood pH and HCO₃^? were evaluated. Blood smears were prepared for the verification of leucocyte profile and neutrophils:lymphocyte ratio (N:L). Blood pCO₂, pH and HCO₃^? increased significantly after transport for all treatments compared with pre‐transport. Glucose levels increased at the higher density whereas no effects were observed on plasma cortisol and pO₂ levels. Upon arrival, all treatments showed lymphopenia and neutrophilia which increased N:L ratio. Although lymphopenia was observed in higher densities until 2 hr after transport, haematological parameters were fully restored within 24 hr post transport. Furthermore, no mortalities were observed throughout the experimental period. Based on the transient physiological changes observed in this study, juvenile dusky grouper can be safely transported in plastic bags for 8 hr at a density of up to 64 g/L.     

Soy protein concentrate as an alternative in replacement of fish meal in the feeds of hybrid grouper,brown‐marbled grouper (Epinephelus fuscoguttatus) × giant grouper (E. lanceolatus) juvenile

Hybrid grouper juveniles (body weight, 6.1 ± 0.7 g) (brown‐marbled grouper, Epinephelus fuscoguttatus × giant grouper, E. lanceolatus) were fed with six isoproteic (50% crude protein) and isolipidic (12% crude lipid) feeds containing different levels of soy protein concentrate (SPC) in replacement of fish meal (SPC at 20%, 30%, 40%, 50% and 60% protein) and control feed (SPC0) for 6 weeks. Hybrid grouper juveniles were cultured in 100‐L fibreglass tank equipped with flow‐through water system and fed twice a day to apparent satiation level. The highest and lowest growth was recorded in fish fed SPC20 and SPC60 respectively. However, growth of SPC20 was not significantly higher than those fed SPC0, SPC30, SPC40 and SPC50 (p > .05). A decreasing growth trend was observed with the increasing level of SPC from feed SPC40 to SPC60. A noticeable better feed utilization was also observed in fish fed SPC0, SPC20, SPC30, SPC40 and SPC50 compared to fish fed SPC60 (p < .05). The fish condition factor, hepatosomatic index, viscerosomatic index and whole body proximate content of the fish were not affected by the graded levels of SPC. However, the body lipid content was significantly lower in fish fed SPC40 to SPC60 (p < .05). The apparent digestibility coefficient (ADC) of protein and lipid was significantly higher in fish fed SPC0 and SPC20 compared to other dietary treatments (p < .05). Based on the regression analysis on specific growth rate, the study suggests that the hybrid grouper grow best at 21.4% and can utilize up to 50% inclusion level of SPC in protein without significantly affect their growth and its body condition.     

Viral encephalopathy and retinopathy is endemic in wild groupers (genus Epinephelus spp.) of the Algerian coast

This work describes betanodavirus infection in two species of groupers (family Serranidae) from the Algerian coast: the dusky grouper Epinephelus marginatus and the golden grouper Epinephelus costae. At necropsy, characteristic clinical signs, external injuries, clouded eyes and brain congestion, generally associated with viral encephalopathy and retinopathy (VER) infection were observed. The partial sequences of RNA1 and RNA2 from two viral strains were obtained, and the phylogenetic analysis revealed the presence of the red-spotted grouper nervous necrosis virus (RGNNV) genotype closely related to strains previously detected in groupers in the same geographic area. Results obtained in this study support the hypothesis that VER disease is endemic in the Algerian grouper population.     

Global population genetic structure of the sequential hermaphrodite,dusky grouper (Epinephelus marginatus)

1. The dusky grouper Epinephelus marginatus is a large epinephelid species that occurs in the eastern and south-western Atlantic and western Indian Oceans. Late maturity, protogynous hermaphroditism, site fidelity, and overfishing have all contributed to its demographic decline.
2. Connectivity and demography within a broad sampling of dusky grouper populations throughout its distribution were assessed. To do so, genetic variation at 11 polymorphic microsatellite loci and a partial sequence of the mitochondrial control region (mtCR) were evaluated.
3. Two major mtCR lineages with a sequence divergence of 1.6% were found. The magnitude of genetic differentiation for mtCR among north and south Atlantic and Indian Ocean populations was high, with Φ_ST = 0.528.
4. D_EST and results of discriminant analysis of principal component revealed significant microsatellite genetic differentiation between all collection areas. Significant pairwise D_EST showed moderate (0.084) to very great (0.603) differentiation. The effective population size was low for all localities, ranging between 25 (Azores Archipelago) and 311 (Rio Grande do Sul). The overall effective population size was estimated as 299 (confidence interval = 215–412), and there was no evidence of strong or recent bottleneck effects.
5. Local and regional genetic structuring among dusky grouper populations is the consequence of the species' site fidelity, distribution across multiple oceanographic boundaries, and probably also of sequential hermaphroditism that contributes to the intensity of random genetic drift.
6. The spatial pattern of genetic structuring of dusky groupers is such that fisheries management and conservation of population genetic integrity will have to be pursued at the local and regional scales.

     

Microbial communities associated with early stages of intensively reared orange‐spotted grouper (Epinephelus coioides)

The gut microbiota plays key roles in the health and general welfare of fish larvae, the present study characterized the bacterial communities associated with grouper Epinephelus coioides larvae during a period of 22 days post hatch (DPH) in an intensive hatchery using both cultivation‐based and cultivation‐independent approaches. Both approaches confirmed that bacteria were present in the gut of larvae before and after the onset of exogenous feeding, and the number of cultiviable bacteria increased gradually from 2 DPH to 22 DPH. A more complex bacterial profile was present in larvae fed fertilizer oyster eggs for 4 days (8 DPH), probably as a result of the onset of exogenous feeding. Interestingly, similar internal microbiota were observed in larvae fed fertilized oyster eggs for 4 days (8 DPH) and rotifers for 2 weeks (22 DPH), although different microbial communities were present in the two feeds. This might suggest that the gut environment of E. coioides larvae selects for a common microbiota, which is more closely related with the rearing water than the two feeds. Therefore, bacterial community of the rearing water may play a critical role in the establishment of gut microbiota of fish larvae and more attention should be paid to its practical modulation by using probiotics. In addition, some potentially beneficial bacteria, such as Lactococcus spp., were the major components of the microbiota associated with fertilized oyster eggs, while these bacteria were not detected in larvae samples.     

Growth performance and disease resistance against Vibrio vulnificus infection of novel hybrid grouper (Epinephelus lanceolatus × Epinephelus fuscoguttatus)

Groupers are economically important for aquaculture in Thailand. A novel hybrid grouper (Epinephelus lanceolatus × Epinephelus fuscoguttatus) has been successful cross‐bred; therefore, the present work aimed to assess the hybrid's traits. The growth performance, strength and tolerance to a pathogenic bacterial infection of this hybrid were compared with its parent species, tiger grouper and giant grouper. The results of all measured growth parameters indicated that the hybrid strain grew fastest followed by giant and tiger grouper respectively. The expressions of the growth‐related genes, insulin‐like growth factor (IGF) I and II, were also analysed in fish muscle and liver which are the main target organs in fish growth regulation. Among tested species, similar expression patterns of IGF‐I and IGF‐II were detected in both organs. The levels of these genes in liver and muscle of hybrid and giant grouper were higher than those of tiger grouper comparable with the growth manner. After challenge with Vibrio vulnificus, the immunological parameters, clearance time of Vibrio in haemolymph and survival was measured to verify the fish immunity. Leucocyte number, lysozyme activity and the ability to eliminate the pathogen were very high in hybrid and giant grouper while these parameters were lower in tiger grouper. Correspondingly, the mortality rate of tiger grouper was higher than others and % survival at the end of observation time (15 days post challenge) was lowest in infected tiger grouper. Altogether, the results suggested that the hybrid grouper has desirable traits that will improve cultured grouper.     

Outbreaks of severe myositis in cultured white sturgeon (Acipenser transmontanus L.) associated with Streptococcus iniae

Outbreaks of an infectious disease affecting cultured white sturgeon (Acipenser transmontanus) were investigated. Clinical signs included erratic swimming, arching of the back and mortality. Necropsy findings included poorly demarcated yellow to dark-red and friable lesions in the epaxial muscle, ulcerative skin lesions and haemorrhages in the swim bladder and coelomic wall. Histological evaluation revealed areas of necrotizing and heterophilic myositis with aggregates of bacterial cocci. The lumen of blood vessels in the dermis, under ulcerated areas, and in the posterior kidney, was occluded by fibrin thrombi. Aggregates of Gram-positive cocci were observed in the muscle lesions and within the fibrin thrombi in the dermis and kidney. Genetically homogeneous Streptococcus iniae strains were recovered from affected fish from different outbreaks. The isolates shared high degree of similarity at gene locus (gyrB) with previously characterized S. iniae from cultured fish in California, confirming the emergence of this particular strain of S. iniae in US aquaculture.     

Effects of dietary inclusion of soybean meal and cholesterol on the growth,cholesterol status and metabolism of the giant grouper (Epinephelus lanceolatus)

The study evaluated effects of cholesterol supplementation in a diet with high soybean meal (SBM) on the growth and cholesterol metabolism of giant grouper (Epinephelus lanceolatus). All‐fish‐meal diet was used as control. The diet including SBM (replaced 50% of the fish meal protein, SBM diet) and the SBM diet supplemented with 10 g/kg cholesterol (SBM + cholesterol) were used as experimental diets. Three diets were each fed to triplicate groups of juvenile grouper (initial body weight: 12.39 ± 0.36 g) in a recirculating aquaculture system for 8 weeks. Grouper fed the control diet showed higher (p < .05) weight gain, feed intake, feed efficiency and protein efficiency ratio than the other two dietary treatments. Hepatic cholesterol concentrations and 3‐hydroxy‐3‐methylglutaryl coenzyme A reductase gene expressions were higher in fish fed the control diet than fish fed the control diet and SBM + cholesterol diet. Hepatic cholesterol 7α‐hydroxylase gene expression was higher in fish fed the SBM + cholesterol diet than that in fish fed the control diet. Results indicate that giant grouper on a diet low in cholesterol can regulate cholesterol synthesis, suggesting that the reduced dietary cholesterol intake in the fish fed diet containing SBM is sufficiently compensated by increased cholesterol synthesis.     

Characterization of triploid hybrid groupers from interspecies hybridization (Epinephelus coioides ♀ × Epinephelus lanceolatus ♂)

Interspecies hybridization is widely used in aquaculture as a beneficial strategy. Diploid and triploid hybrids have been detected from the interspecies hybridization of Epinephelus coioides ♀ × Epinephelus lanceolatus ♂. This is the first report of triploidization through hybridization in grouper. Confirmation has been obtained through flow cytometry, karyotyping and erythrocyte nuclei measurement. The chromosome numbers of E. coioides, E. lanceolatus, diploid hybrid grouper are 48 and triploid hybrid grouper are 72. Measurements of erythrocyte nuclei indicate that triploid fish have a larger nuclear surface than the diploid groupers, and the average ratio of triploid to diploid surface area is 1.59. During the first 1.5 years, triploid hybrid groupers grow faster than diploid hybrid groupers or either parent species. The average growth rate of triploid hybrids is 1.61 times greater than that of diploid hybrids at 6 months of age and 1.43 times greater at 18 months of age. The triploid hybrid groupers are inferior in gonadal development, with no primary‐growth‐stage oocytes appearing in the gonads at 18 months of age. Morphological studies indicate that triploid hybrid groupers have distinctive differences in snout length, eye diameter, body trunk shape, and tail shape development compared with diploid hybrid groupers. Triploid hybrid groupers have an advantage in growth ability, and artificial breeding of triploid groupers might be of great potential use in the grouper aquaculture industry.     

Granulomatous lesions in a wild mullet population from the eastern Ligurian Sea (Italy): mycobacteriosis vs. pseudotuberculosis

Mycobacterium spp. and Photobacterium damselae subsp. piscicida are recognized as the most frequent causative agents of granulomatous lesions in fish. Although frequent episodes of mycobacterial infections have been reported in wild fish worldwide, only sporadic cases have been documented to date in Italy. To investigate for the presence of lesions referable to mycobacteriosis and to identify the mycobacterial species involved, a total of 159 wild mullets were fished from the eastern coast of the Ligurian Sea, killed and necropsied. Liver and spleen samples were collected from all fish for histopathological and microbiological analyses. Molecular investigations for identification of Photobacterium damselae subsp. piscicida were performed. Gross examination revealed granulomatous lesions in one animal; microscopically, 42.14% of fish displayed granulomas with various histological features, 19.50% resulted positive at Ziehl–Neelsen staining, and were confirmed as mycobacterial lesions by culture. The identified colonies were characterized as M. fortuitum, M. abscessus, M. flavescens, M. chelonae, M. septicum and M. nonchromogenicum. In all, 35% of animals resulted positive for Photobacterium damselae subsp. piscicida. These data suggest widespread mycobacterial infection also by Photobacterium damselae subsp. piscicida infections in wild fish. Moreover, the pathogenicity of some mycobacterial species, previously considered as saprophytic, was demonstrated.