Red mark syndrome of trout (Oncorhynchus mykiss; Walbaum, 1792): Histopathological scoring and correlation with gross lesions.

Red mark syndrome (RMS) is a skin disorder affecting rainbow trout (Oncorhynchus mykiss). The present work aimed to correlate the gross skin lesions affecting 46 fish sampled from farms surveyed for RMS with their microscopic features, identifying histological parameters that may be suggestive of disease progression. Skin lesions were grossly included in one of three categories (types I, II and III) according to the progressive degree of severity. Histological parameters and anti-proliferating cell nuclear antigen (PCNA) tissue immunoreactivity were semi-quantitatively assessed. In the dermis, PCNA-positive lymphocytes, fibroblasts and endothelial cells were indicative of active phlogosis. A significant increase in PCNA-immunoreactive lymphocytes, from gross type I to type III cases, was found only in the hypodermis. The histological parameters significantly associated with the gross lesion severity were progressive loss of the epithelium and scales, recruitment of inflammatory cells in the stratum compactum, loss of architecture of the stratum compactum, perivascular and perineural granulomatous inflammation and increase in lymphocyte infiltration of the muscular layer. In the type II and type III categories, inflammation in the hypodermis and muscle displayed a granulomatous pattern, reinforcing the hypothesis of an immunopathological mechanism. The morphological diagnosis of "deep chronic dermatitis associated to panniculitis and myositis, characterised by lympho-histiocytic and granulomatous reaction" is suggested.

Effects of feeding low fishmeal diets with increasing soybean meal levels on growth, gut histology and plasma biochemistry of sea bass.

The aquaculture industry depends upon the development of sustainable protein sources to replace fishmeal (FM) in aquafeeds and the products derived from soybeans are some of the most studied plant feedstuffs. A key area of investigation for continuing to improve modern aquafeeds includes the evaluation of varying proportions and combinations of plant ingredients to identify mixtures that are more efficiently utilized by the fish. This study investigated the effects of increasing soybean meal (SBM) by replacing a mix of plant ingredients in low FM (20%) diets on growth, blood biochemistry profile and gut histology on European sea bass. Five isonitrogenous and isolipidic experimental diets were formulated: four diets containing increasing SBM levels (0, 10, 20 and 30%; 0SBM, 10SBM, 20SBM and 30SBM, respectively) with a low content of FM (20%) and one control diet (0% SBM; 35% FM). Diets containing SBM brought to comparable performance and protein utilization, while 0SBM had negative impact on feed conversion rate and protein utilization. Blood parameters suggested an optimal nutritional status under all feeding treatments, even though slightly decreased values were reported at increasing dietary SBM. Histology examination did not show any changes indicative of soy-induced enteritis. We can conclude that for European sea bass: (i) different blends of plant protein did not affect feed intake despite the 20% FM dietary level; (ii) the inclusion of SBM maintains optimal growth and feed utilization in low FM diets; (iii) blood biochemistry profile showed a good nutritional status under all feeding regimes; (iv) no evidence of soy-induced enteritis was reported in any group fed low FM diets. For formulation of practical diets in on-growing of European sea bass, SBM up to 30% can be successfully incorporated into feeds containing low FM inclusion.

Outbreak of mortality in Russian (Acipenser gueldenstaedtii) and Siberian (Acipenser baerii) sturgeons associated with sturgeon nucleo-cytoplasmatic large DNA virus.

Diseased outbreaks with high mortality in farmed sturgeon are a limiting factor to the success of this emerging aquaculture sector in Europe. Thorough investigations of outbreaks can determine the aetiological agents, identify important pathological and epidemiological pathways of infections and pave the way for effective control strategies. A thorough investigation of a mortality outbreak in Russian (Acipenser gueldenstaedtii) and Siberian (Acipenser baerii) sturgeons in Italy, demonstrated the primary involvement of a sturgeon nucleo-cytoplasmic large DNA virus (NCLDV). While, the taxonomy classification of this new virus is still uncertain, its involvement in sturgeon mortality outbreaks in Europe is, for the first time, fully investigated and described. Furthermore, the coinfection of bacteria such as motile Aeromonas spp. and Acinetobacter spp. was reported. Genetic characterisation showed the close relationship between the European sturgeon NCLDV with North American sturgeon NCLDVs. Similarly to the latter, the European sturgeon NCLDV persists in survivors. Furthermore, a systemic distribution of the European sturgeon NCLDV was evident in diseased A. baerii and A. gueldenstaedtii and in recovered A. gueldenstaedtii. These epidemiological and pathological findings will help in the identification of effective control strategies for sturgeon NCLDV infection, which afflicts an important and emerging European aquaculture sector.

Rhabdomyosarcoma of Soft Tissues in an Adult Brook Trout (Salvelinus fontinalis).

This report describes a spontaneously arising rhabdomyosarcoma of soft tissues in a brook trout (Salvelinus fontinalis). The lesion was examined by means of histology, immunohistochemistry (IHC) and transmission electron microscopy (TEM). The cross-reactivity of the primary antibodies used in the IHC was investigated in silico using the Protein Blast system. Microscopically, the lesion appeared as a 'small round cell' undifferentiated sarcoma with rare myotube formation. IHC identified expression of sarcomeric actin and vimentin and these molecules showed the highest protein sequence identity. Lower protein sequence identity coincided with negative immunolabelling for desmin, MyoD1, myogenin and CD3. TEM revealed myofibrils, but without a defined sarcomeric architecture. The diagnosis of solid alveolar rhabdomyosarcoma of soft tissues was achieved on the basis of histological and ultrastructural findings.

Effect of photoperiod on endocrine profiles and vitellogenin expression in European eels Anguilla anguilla during artificially induced ovarian development.

The aim of this work was to determine the effects of dark and light conditions on the E2, testosterone and thyroid hormones levels and on the gene expression levels (vitellogenin 1, vitellogenin 2, and estradiol receptor one) in European eels (Anguilla anguilla) during ovarian development induced by increasing doses of carp pituitary extracts (CPEs). The subjects were divided into 2 groups: 14-hour light:10-hour dark (Light Group) and 24-hour darkness (Dark Group). All the eels received intramuscular injections with CPE at a dosage of 10 mg/kg body weight (BW) once a week for the first 3 weeks, 20 mg/kg BW fourth-sixth week, 30 mg/kg BW seventh-ninth week, and 40 mg/kg up to the end of the experiment (13th week). Vitellogenin and estradiol receptor expression levels did not show significant differences between the two housing conditions whereas in both groups vitellogenin mRNA increased starting from first CPE injection. Testosterone and 17-beta estradiol plasma levels were significantly greater in the Dark Group compared with the Light Group starting from the ninth and the 13th week, respectively. These results suggest that darkness could be a useful variable for standardizing gonadal maturation in eels kept in captivity.

Enteric neuroplasticity in seawater-adapted European eel (Anguilla anguilla).

European eels live most of their lives in freshwater until spawning migration to the Sargasso Sea. During seawater adaptation, eels modify their physiology, and their digestive system adapts to the new environment, drinking salt water to compensate for the continuous water loss. In that period, eels stop feeding until spawning. Thus, the eel represents a unique model to understand the adaptive changes of the enteric nervous system (ENS) to modified salinity and starvation. To this purpose, we assessed and compared the enteric neuronal density in the cranial portion of the intestine of freshwater eels (control), lagoon eels captured in brackish water before their migration to the Sargasso Sea (T0), and starved seawater eels hormonally induced to sexual maturity (T18; 18 weeks of starvation and treatment with standardized carp pituitary extract). Furthermore, we analyzed the modification of intestinal neuronal density of hormonally untreated eels during prolonged starvation (10 weeks) in seawater and freshwater. The density of myenteric (MP) and submucosal plexus (SMP) HuC/D-immunoreactive (Hu-IR) neurons was assessed in wholemount preparations and cryosections. The number of MP and SMP HuC/D-IR neurons progressively increased from the freshwater to the salty water habitat (control > T0 > T18; P < 0.05). Compared with freshwater eels, the number of MP and SMP HuC/D-IR neurons significantly increased (P < 0.05) in the intestine of starved untreated salt water eels. In conclusion, high salinity evokes enteric neuroplasticity as indicated by the increasing number of HuC/D-IR MP and SMP neurons, a mechanism likely contributing to maintaining the body homeostasis of this fish in extreme conditions.

Effects of two water disinfectants (chloramine T and peracetic acid) on the epidermis and gills of Garra rufa used in human ichthyotherapy.

Doctor fish (Garra rufa) have recently been used for aesthetic purposes and as a medical treatment in patients with psoriasis (ichthyotherapy). For this particular kind of human therapy it is essential to guarantee adequate hygienic conditions for both people and fish. The aim of this study was to test two concentrations of water disinfectants, chloramine T and peracetic acid, on Garra rufa to ascertain possible exposure damage to the epidermis and gills. Fish were exposed to 2 mg/l and 10 mg/l of chloramine T and to 15 microl/l and 45 microl/l of peracetic acid in a 40-minute static bath up to six times a day for one week. The epidermis and gills were checked for histological changes and the number of epidermal mucous cells, club cells and taste buds were quantified; mucous cells were also characterized histochemically to detect alterations in mucin production. No mortality or severe histological changes were found in treated or control fish. Cell count showed a significant increase (p < 0.05) in mucous cells (mean 49.1 +/- 6.7 vs 37.0 +/- 13.1 of controls) in animals treated with peracetic acid independently of the dose. Club cell number showed a significant (p < 0.05) decrease in fish treated with 2 mg/l of chloramine T (mean 74.3 +/- 15.6) and with 45 microl/1 of peracetic acid (mean 78.17 +/- 10.5) compared to controls (mean 107.0 +/- 19.2). Histochemical evaluation of mucous cells did not reveal changes in mucin type in fish exposed to the two disinfectants. The results suggest a good tolerability of Garra rufa to the two disinfectants at the concentrations tested.

Thyroid hormones as a potential early biomarker of exposure to 4-nonylphenol in adult male shubunkins (Carassius auratus).

Nonylphenol, an estrogenic-like compound, can induce vitellogenin synthesis in males and immature Teleostean species, but little is known about its effects on thyroid hormones balance. The present study evaluated the potential effects of a single acute exposure to nonylphenol (i.p. injected) on the thyroid and reproductive axis of 250 shubunkins (Carassius auratus). Plasma levels of thyroid hormones were quantified immunoenzymatically by ELISA assay. Nonylphenol induced a significant decrease of thyroxin levels, whereas no effect on triiodothyronine concentrations was detected. No histopathological changes were detected in thyroid or testes. The toxicological data confirmed that nonylphenol exerts an estrogenic effect on male fish. In addition, nonylphenol was suspected to inhibit the thyroid hormones balance, suggesting the thyroid should be included among the other endocrine glands susceptible to endocrine disruption.