RESUMO
The aim of this study was to assess the effects of low levels of dietary fish meal (FM) and fish oil (FO) on disease resistance and gut associated lymphoid tissue (GALT) response after an experimental intestinal infection with V. anguillarum in European sea bass (Dicentrarchus labrax) For that purpose, sea bass juveniles were fed one of four diets containing combined levels of FO and FM as follows: 20%FM/6%FO, 20%FM/3%FO, 5%FM/6%FO and 5%FM/3%FO during 153 days. At the end of the feeding trial, fish were subjected to either an in vivo exposure to a sub-lethal dose of V. anguillarum via anal inoculation or to an ex vivo exposure to V. anguillarum. Additionally, inducible nitric oxide synthase (iNOS) and tumor necrosis factor α (TNFα) gut patterns of immunopositivity were studied. Growth performance was affected by dietary FM level, however ex vivo gut bacterial translocation rates and survival after the in vivo challenge test were affected by dietary FO level. After 5 months of feeding, low dietary FM levels led to a posterior gut up-regulation of interleukin-1ß (IL-1ß) and TNFα, major histocompatibility complex-II (MHCII) and cyclooxygenase-2 (COX2), which in turn reduced the gut associated lymphoid tissue (GALT) capacity of response after 24 h post infection and conditioned European sea bass capacity to recover gut homeostasis 7 days post infection. Immunoreactivity to anti-iNOS and anti-TNFα presented a gradient of increased immunopositivity towards the anus, regardless of the dietary FM/FO fed. Strong positive anti-TNFα isolated enterocytes were observed in the anterior gut in relation to low levels of dietary FM/FO. Submucosa and lamina propria immunoreactivity grade was related to the amount of leucocyte populations infiltrated and goblet cells presented immunopositivity to anti-iNOS but not to anti-TNFα. Thus, reducing FO content from 6% to a 3% by VO in European sea bass diets increases ex vivo and in vivo gut bacterial translocation rates, whereas reducing FM content from 20% down to 5% up-regulates the expression of several posterior gut inflammation-related genes conditioning fish growth and GALT capacity of response after bacterial infection.
Assuntos
Bass/imunologia , Suplementos Nutricionais , Resistência à Doença , Doenças dos Peixes/imunologia , Óleos de Peixe/imunologia , Enteropatias/veterinária , Vibrioses/veterinária , Ração Animal , Animais , Dieta/veterinária , Enteropatias/imunologia , Vibrio/fisiologia , Vibrioses/imunologiaRESUMO
Plant ingredients and processed animal proteins are alternative feedstuffs for fish feeds in aquaculture. However, their use can introduce contaminants like pesticides that are not previously associated with marine Atlantic salmon and gilthead sea bream farming. This study covers the screening of around 800 pesticides by gas chromatography (GC) and liquid chromatography (LC) coupled to high resolution time-of-flight mass spectrometry in matrices throughout the entire marine food production chain. Prior to analysis of real-world samples, the screening methodology was validated for 252 pesticides to establish the screening detection limit. This was 0.01 mg kg-1 for 113 pesticides (45%), 0.05 mg kg-1 for 73 pesticides (29%) and >0.05 mg kg-1 for 66 pesticides (26%). After that, a quantitative methodology based on GC coupled to tandem mass spectrometry with atmospheric pressure chemical ionization source (GC-APCI-MS/MS) was optimized for the pesticides found in the screening. Although several polar pesticides, of which pirimiphos methyl and chlorpyriphos-methyl were most dominant, were found in plant material and feeds based on these ingredients, none of them were observed in fillets of Atlantic salmon and gilthead sea bream fed on these feeds.