Oral immunization of trout fry with recombinant Lactococcus lactis NZ3900 expressing G gene of viral hemorrhagic septicaemia virus (VHSV).

This study has investigated the ability of Lactococcus lactic (NZ3900) carried G gene of viral haemorrhagic septicaemia virus (VHSV) under nisin-controlled gene expression (NICE) system in rainbow trout (O.Mykiss). Two groups of trout fry (7 ± 0.65 g) were immunized with 1 × 1010 cfu/g and 1 × 108 cfu/g recombinant L. lactis NZ3900, two groups of fish were fed 1 × 1010 cfu/g and 1 × 108 cfu/g L. lactis vector free, and one group was fed by the basal diet as a control. Oral immunization was done on days 1-7 and boosting was performed on days 15-21. The relative expression of IFN-1 and MX-1 genes significantly increased in head kidney of vaccinated fish depend on vaccine dosage compared to the control group. Fish in vaccinated group also showed elevated VHSV-specific antibody levels compared to the control groups. Relative percent survival (RPS), under virulent isolate VHSV challenge were estimated 62%, 78% for 108 cfu/g 1010 cfu/g feed vaccinated groups 21 days post-vaccination, while groups fed similar doses of L. lactis vector free illustrated 22% and 27% RPSs, respectively. The significant reduction of viral loads (transcript levels of N gene) were detected in the immunized groups. Increased weight gain and decreased feed consumption in vaccinated group attributed to the probiotic effect were also observed. In conclusion, our results demonstrate the ability of recombinant L. lactis as oral vaccine against VHS in rainbow trout, which can be considered as effective method against different fish pathogens.

Dietary Immunogen® modulated digestive enzyme activity and immune gene expression in Litopenaeus vannamei post larvae.

Pacific white shrimp Litopenaeus vannamei (Boone, 1931) is an important economical shrimp species worldwide, especially in the Middle East region, and farming activities of this species have been largely affected by diseases, mostly viral and bacterial diseases. Scientists have started to use prebiotics for bolstering the immune status of the animal. This study aimed to investigate the influence of Immunogen® on growth, digestive enzyme activity and immune related gene expression of Litopenaeus vannamei post-larvae. All post-larvae were acclimated to the laboratory condition for 14 days. Upon acclimation, shrimps were fed on different levels of Immunogen® (0, 0.5, 1 and 1.5 g kg-1) for 60 days. No significant differences were detected in weight gain, specific growth rate (SGR) and food conversion ratio (FCR) in shrimp post-larvae in which fed with different levels of Immunogen® and control diet. The results showed that digestive enzymes activity including protease and lipase increased with different amounts of Immunogen® in the shrimp diet. Protease activity increased with 1.5 g kg-1 Immunogen® after 60 days and lipase activity increased with 1 and 1.5 g kg-1 Immunogen® after 30 and 60 days of the trial respectively (P < 0.05), while amylase activity did not change in response to different levels of Immunogen® (P > 0.05). The expression of immune related genes including, prophenoloxidase, crustin and g-type lysozyme increased with diet 1.5 g kg-1 Immunogen® (P < 0.05) while expression of penaeidin gene increased only with experimental diet 1 g kg-1 of Immunogen®. These results indicated that increase in digestive enzymes activity and expression of immune related genes could modulate the Immunogen® in the innate immune system in L. vannamei in this study.

Effects of (Anti) Androgenic Endocrine Disruptors (DEHP and Butachlor) on Immunoglobulin M (IgM) and Leukocytes Counts of Male Rainbow Trout (Oncorhynchus mykiss).

The effect of two anti-androgenic endocrine disrupting compounds, i.e. the plasticizer di (2-ethylhexyl) phthalate (DEHP) and herbicide butachlor, were evaluated for their effects on immunoglobulin M (IgM) and leukocytes in male rainbow trout. Also, plasma testosterone (T) concentration was measured to confirm their anti-androgenic effects. In the first experiment, trout were treated with 50 mg/kg (body weight) DEHP intraperitoneally, and in the second one, fish were exposed to 0.39 mg/L butachlor for 10 days. The results showed that T concentrations and white blood cells were significantly lower in fish exposed to either DEHP or butachlor compared to control fish (p < 0.05). Fish showed significantly elevated neutrophil levels and decreased lymphocyte levels in the butachlor (p < 0.05); however, no significant difference was observed in lymphocyte and neutrophils values in the DEHP treatment (p > 0.05). In addition, no significant differences were found in IgM, eosinophil and monocyte parameters in either DEHP or butachlor treatments (p > 0.05). These results confirmed that leukocytes counts can be considered as a novel marker of immunotoxicity triggered by (anti) androgenic endocrine disruptors.

Morphological study of the northern pike (Esox lucius) tongue.

The northern pike (Esox lucius) is a fresh water species belonging to the Esocidae family. It is a carnivorous fish feeding mostly on invertebrates and fishes. Due to the scantiness of relevant literature regarding the morphology of the tongue in fish we carried out this study with the aim of providing information on the dorsal surface morphology and histological structures of the tongue in E. lucius. The tongues of five E. lucius were examined using light- and scanning electron- microscopy (SEM) techniques. The SEM studies revealed the presence of numerous teeth, longitudinal mucosal strands and scattered taste buds spread on the tongue surface. Histological studies using hematoxylin and eosin and Masson's trichrome staining showed that the musculature was not visible in the tongue of E. lucius. The tongue is composed of mucosa, and submucosa supported by osteocartilagionous skeleton. The mucosa consists of several layers of unicellular mucous cells interrupted by numerous teeth. The derivation of teeth from the underlying bronchial skeleton was visible in longitudinal section. The scattered taste buds with a typical onion shape were also present. Overall, the morphological features of the E. lucius tongue together suggested its mechanical and sensory roles. The findings of this study together with morphological and physiological data from other fishes contribute to the knowledge of the nutrition and feeding behavior in aquaculture species.