One of the main objectives of the present study was an effective replacement of dietary fishmeal/fish oil (FM/FO) by new raw materials without negatively affecting European sea bass (Dicentrarchus labrax) performance and health status within a selection breeding context. The genomic selection of this species is still in its infancy if compared to salmonids and in particular, the role of D.labrax genome in shaping the gut microbiome, has been scarcely investigated. Wildtype (WT) and selected (GS) sea bass were fed with two diets: a control (20% FM/ 5-9% FO), and a "future" (F) diet, in which FM was decreased to 10% being replaced by poultry meal, whereas FO was completely replaced by a blend of rapeseed, poultry, and algae oils. The morphological evaluation of the intestine revealed a well-organized folding pattern and a conserved gut epithelial barrier for all fish groups. Despite a basal level of inflammation in the proximal intestine of WT fish, no differences were observed neither in the morphometric characteristics of goblet cells nor in the expression of GALT-related genes in response to fish genotype or diet. At distal intestine, WT fish showed a higher inflammatory status and larger goblet cells than GS fish and within the same genotype, fish fed the F diet had in general larger goblet cells. In distal intestine, a significant effect was found on the expression of 3 out of 7 target GALT genes. In particular, the expression of cytokines il-1 beta, tnf-alpha, and il-10 was different, showing an interaction effect diet x genotype. Diet had a lower influence upon gut bacterial composition than genotype. Indeed, regardless of the diet, WT fish showed higher species richness than GS genotype and this could be a direct consequence of selective breeding for multiple traits selection including growth, external morphology for lower abdominal fat deposition and adaptation to multiple and successive feed sources and composition across generations of selection. Furthermore, the gut microbiota of GS fish shared a reduced individual variability, indicating an enhanced capacity to cope with changes in diet composition. The less changes of GS sea bass at the level of gut bacterial composition in cumulating data collected with the feeds, demonstrate a capacity to reshape their microbiota thus better adapting to the diet, but with no negative impact on their growth performances, and even a better growth. A significant genotype effect was found for specific bacterial taxa, such as Paracoccus genus and other genera belonging to Moraxellaceae family, which were enriched in WT fish, regardless of the diet. Interestingly, the relative abundance of Paracoccus genus was positively correlated with higher proinflammatory cytokine il-1 beta expression found in distal intestine of wildtype sea bass.

The sustainable development of the aquaculture industry relies on the use of alternative conventional and emergent raw materials that contributes to a circular economy and to reduce the dependency on fish meals and fish oils coming from oceanic fish populations. Additionally, the genetic selection of farmed fish that can display higher growth and feed utilization when fed alternative feeds, is pointed out to be a complementary valuable tool to facilitate the implementation of circular economy approaches. The main purpose of the present study was to determine the effectiveness of genetic selection for growth in European sea bass, in response to a challenge with an alternative diet that aimed to partially replaced fishmeal (FM) by poultry meal (PM) and totally replace fish oil (FO) by a blend of poultry oil (PO) with a novel microalgae oil. The two families of fish juveniles were obtained by in vitro fertilization of selected for a multi-trait including high growth (genetically selected, GS) or non-selected (wild type, WT) broodstocks and then were nutritionally challenged with a control diet that mirrored a standard commercial diet with fishmeal (20%) and fish oil (7%), or a Future diet that partially replaced the FM by PM and totally replaced the FO by a blend of rapeseed oil, PO, and a novel DHA rich-algal oil. From the second month of feeding until the end of the trial, European sea bass that was selected since for 7 generations performed better in terms of growth than the wild-type genotype, possibly related with an apparent favored feed and nutrient utilization. Furthermore, selection decreased the perivisceral fat and increased the nutritional value of flesh by increasing DHA (in g/ 100 g flesh) and ARA contents. In contrast, the dietary treatment showed little effect on fish growth performance, denoting the successful partial replacement of FM by PM and the total replacement of FO by a blend of poultry oil and an emergent microalgal oil. However, Future diet tended to reduce the ADCs of some amino acids, as well as showed an additive effect to genotype in increasing the n-3 PUFA of flesh. Altogether, our data demonstrate that multi-trait genetic selection of European sea bass improve fish plasticity to cope with the variations of ingredients in alternative feeds with low FM/FO.

Peracetic acid (PAA) is a European Union Commission authorized disinfectant for use in animal health care. It has shown a strong inactivation potential for bacteria, viruses, fungi and bacterial spores. A stress-related adaptive response after an exposure to PAA has been described in different species of fish such as carp (Cyprinus carpio), rainbow trout (Oncorhynchus mykiss). The present study aims to evaluate the response of gilthead seabream systemic (plasma) as mucosal (gill and skin) defences after PAA a unique exposure (5 min), by measuring cortisol, glucose and lactate in plasma, as well as the expression of several genes (glutathione peroxidase, glucocorticoid receptor, superoxide dismutase 2 and superoxide dismutase involved in the response to oxidative stress in mucosal tissues). study. We observe how seabream (Sparus aurata) copes with oxidative stress induced by PAA. PAA exposure did not induce an important antioxidant response in fish, whereas induced a mild response to stress, with a fast and effective recovery of basal levels after 24 h. Although PAA triggers a mild stress response, the response described in our study reflect that it can be used for sea bream in the concentration tested with no severe physiological consequences.

The use of terrestrial raw materials to replace fish meal (FM) and fish oil (FO) in marine fish diets may affect fish growth performance and health. In the last years functional additives have been profiled as good candidates to reduce the effects on health and disease resistance derived from this replacement, via reinforcement of the fish immune system. In the present study, three isoenergetic and isonitrogenous diets with low FM and FO (10% and 6% respectively) were tested based on supplementation either with 0.5% galactomannanoligosaccharides (GMOS diet) or 0.02% of a mixture of essential oils (PHYTO diet), a non-supplemented diet was defined as a control diet. Fish were fed the experimental diets in triplicate for 9 weeks and then they were subjected to a stress by confinement as a single challenge (C treatment) or combined with an experimental intestinal infection with Vibrio anguillarum (CI treatment). Along the challenge test, selected stress and immunological parameters were evaluated at 2, 24 and 168h after C or CI challenges. As stress indicators, circulating plasma cortisol and glucose concentrations were analyzed as well as the relative gene expression of cyp11b hydroxylase, hypoxia inducible factor, steroidogenic acute regulatory protein, heat shock protein 70 and heat shock protein 90 (cyp11b, hif-1 alpha, StAR, hsp70 and hsp90). As immune markers, serum and skin mucus lysozyme, bactericidal and peroxidase activities were measured, as well as gene expression of Caspase-3 (carp-3) and interleukin 1 beta (il-1 beta). The use of functional additives induced a significant (p < 0.05) reduction of circulating plasma cortisol concentration when confinement was the unique challenge test applied. Supplementation of PHYTO induced a down-regulation of cyp11b, hif-1 alpha, carp-3 and il-1 beta gene expression 2h after stress test, whereas StAR expression was significantly (p < 0.05) up-regulated. However, when combination of confinement stress and infection was applied (CI treatment), the use of PHYTO significantly (p < 0.05) down-regulated StAR and carp-3 gene expression 2h after challenge test, denoting that PHYTO diet reinforced fish capacity of stress response via protection of head kidney leucocytes from stress-related apoptotic processes, with lower caspase-3 gene expression and a higher il-1 beta gene expression when an infection occurs. Additionally, dietary supplementation with GMOS and PHYTO compounds increased fish serum lysozyme after infection. Both functional additives entailed a better capability of the animals to cope with infection in European sea bass when fed low FM and FO diets.

Neobenedenia girellae is considered an epizootic infection for intensively cultured fish species. Particularly, for greater amberjack (Seriola dumerili) N. girellae causes high mortality rates and supposes a bottleneck during its on-growing period. Thus, the objective of this work was to describe the skin morphological alterations caused by a N. girellae infection on greater amberjack. Greater amberjack juveniles were sampled pre and post experimental infection with N. girellae obtaining cranial and dorsal skin samples. Samples were processed for morphological and ultrastructural studies and revealed clear differences in the structure of both regions, confirming the cranial region as the most susceptible region to be parasitized due to an absence of scales and lower goblet cells density. N. girellae adhesion disrupted the structure of epidermal epithelial cells by overpressure. Stratum spongiosum surface-epithelial cells located near the parasite presented a clear cell degradation process, associated in some cases with cellular detachment. N. girellae infection induced epidermal hydropic degeneration and, in some cases, focal spongiosis. Tissue ulcerative lesions caused by the parasite's attachment structures were characterized by a specific mobilization of leucocytes to the fixation areas. Thus, N.girellae induces important alterations in greater amberjack epidermis independently of the skin region that explain the appearance of secondary infections and associated mortalities.

The main objective of this study was to evaluate the stress response of the greater amberjack, Seriola dumerili, by determining plasma and skin mucus cortisol levels, as physiological indicator of fish stress, and skin mucin-2 gene expression as indicator of skin mucin production, under three different potential stressors related to aquaculture practices: temperature, handling and fasting. For trial 1, 45 greater amberjack juveniles (19.5 +/- 4.1 g) were divided in three graded rearing temperatures (17, 22 and 26 degrees C) and maintained for 120 days, in order to define the basal cortisol level for this species after a long-term thermal acclimation within the range described for this species. Trial 2 was conducted under two different rearing densities and two different handling protocols with 222 fish (425.02 +/- 36.9 g) divided in 4 treatments by triplicate for 90 days, in order to study the effect of routine handling protocols on the selected stress markers for this species. Trial 3 used 72 fish (302.27 +/- 21.4 g) divided in two experimental groups, one fed to apparent satiety and another one subjected to fasting for 10 days (triplicates for each group). After trials 2 and 3, a shallow water challenge was conducted to further study the adaptive response of greater amberjack juveniles to combined stress situations. Plasma and skin mucus were obtained for cortisol determination at 1, 24 and 72 h (trial 2) and 1, 3 and 24 h (trial 3) post stress challenge. Additionally, skin was dissected out for determining muc-2 gene expression. Longterm acclimation to temperature did not affect plasma or mucus cortisol levels and muc-2 gene expression, being basal levels of circulating plasma cortisol around 7.5 ng/mL. A high stocking density (8 kg m(-3)) induced an elevation (p < .05) of plasma cortisol and a reduction (p < .05) of weight gain. No differences were observed for the expression of muc-2 gene, however, a tendency to an up-regulation was observed in fish subjected to the high-density rearing treatment. Short-term fasting induced an increase (p < .05) of plasma and skin mucus cortisol together with a down-regulation of muc-2 gene expression.

The main objective of this study was to determine the effect of two feeding additives (cMOS: Actigen (R), Alltech Inc., USA) and a phytogenic specifically designed for reducing ectoparasite incidence, combining garlic and labiatae herbal extracts (HERB; Delacon, Austria) on growth performance, stress and immune response of greater amberjack (Seriola dumerili) before and after an experimental parasitization with the skin ectoparasite Neobenedenia girellae, a monogenean with high incidence on Seriola cultured in sea cages. Two parallel trials were conducted in order to determine the effect of 70 days of functional diets supplementation (0.2% cMOS and 200 ppm HERB) on greater amberjack: a) Growth performance and stress resistance and b) disease resistance against N. girellae. Additionally, in order to evaluate the effects of the dietary supplements preand postparasitization challenge against N. girellae, the expression of some immune-related genes was evaluated. The relative expression of Il-1 beta, piscidin, hepcidin, IgT, tnf-alpha, muc-2, cathelicidin, complement c3, cd8, and caspase-3 was determined in skin. The activity of protease and lysozyme in serum and skin mucus were also evaluated, as well as parasite load and parasite total size. The use of both functional additives did not affect fish growth performance. However, the dietary supplementation of HERB significantly (p<0.05) reduced circulating cortisol levels after the stress challenge, whereas dietary cMOS significantly (p<0.05) reduced the parasite load and total parasite size. When comparing pre and post parasitization challenge, skin mucus protease activity was significantly (p<0.05) higher in all the treatments before parasitization, while the use of both functional additives increased skin mucus lysozyme activity after challenge. From the eleven immune genes studied, only piscidin was significantly (p<0.05) upregulated by HERB supplementation before challenge, whereas after parasitization, muc-2 and piscidin were significantly (p<0.05) up-regulated by HERB and tnf-alpha, il1-beta, hep, c3, cd8 and casp3 were up-regulated by both functional additives. Moreover, hep, IgT, cath, c3 and cd8 expressions were negatively correlated with the parasite load. In general, greater amberjack supplementation with cMOS and HERB at the dose used in the present study was effective for reducing stress and for increasing the skin associated lymphoid tissue (SALT) immune response against N. girellae during the infection process.

The use of terrestrial raw materials to replace fish meal (FM) and fish oil (FO) in marine fish diets may affect fish growth performance and health. In the last years functional additives have been profiled as good candidates to reduce the effects on health and disease resistance derived from this replacement, via reinforcement of the fish immune system. In the present study, three isoenergetic and isonitrogenous diets with low FM and FO (10% and 6% respectively) were tested based on supplementation either with 0.5% galactomannanoligosaccharides (GMOS diet) or 0.02% of a mixture of essential oils (PHYTO diet), a non-supplemented diet was defined as a control diet. Fish were fed the experimental diets in triplicate for 9 weeks and then they were subjected to a stress by confinement as a single challenge (C treatment) or combined with an experimental intestinal infection with Vibrio anguillarum (CI treatment). Along the challenge test, selected stress and immunological parameters were evaluated at 2, 24 and 168h after C or CI challenges. As stress indicators, circulating plasma cortisol and glucose concentrations were analyzed as well as the relative gene expression of cyp11b hydroxylase, hypoxia inducible factor, steroidogenic acute regulatory protein, heat shock protein 70 and heat shock protein 90 (cyp11b, hif-1α, StAR, hsp70 and hsp90). As immune markers, serum and skin mucus lysozyme, bactericidal and peroxidase activities were measured, as well as gene expression of Caspase-3 (casp-3) and interleukin 1β (il-1ß).The use of functional additives induced a significant (p < 0.05) reduction of circulating plasma cortisol concentration when confinement was the unique challenge test applied. Supplementation of PHYTO induced a down-regulation of cyp11b, hif-1α, casp-3 and il-1β gene expression 2h after stress test, whereas StAR expression was significantly (p < 0.05) up-regulated. However, when combination of confinement stress and infection was applied (CI treatment), the use of PHYTO significantly (p < 0.05) down-regulated StAR and casp-3 gene expression 2h after challenge test, denoting that PHYTO diet reinforced fish capacity of stress response via protection of head kidney leucocytes from stress-related apoptotic processes, with lower caspase-3 gene expression and a higher il-1β gene expression when an infection occurs. Additionally, dietary supplementation with GMOS and PHYTO compounds increased fish serum lysozyme after infection. Both functional additives entailed a better capability of the animals to cope with infection in European sea bass when fed low FM and FO diets.

Vegetable oils (VO) are being extensively used as lipid sources in marine fish diets, due to the decreased availability of fish oil. The use of VO could induce some imbalances on essential fatty acids (EFAs) with the subsequent well described detrimental effects on fish immunity. EFAs are directly associated with several immune functions, acting as important elements for the fluidity of cytoplasmic membranes and as precursors of eicosanoids. At intestinal level, eicosanoids produced via cyclooxygenase (COX-2) enzymes are involved in gastrointestinal cyto-protection by influencing the homeostasis regulation and the onset of gut inflammation process through the action on eicosanoid receptors (EPs), which modulate gastrointestinal mucosal integrity amongst other functions. Senegalese sole is a flatfish with high susceptibility to deleterious effects due to stress and diseases and highly sensitive to changes in dietary fatty acids. In this study, Senegalese sole juveniles were fed diets (56% crude protein, 12% crude lipid) containing linseed (100LO), soybean (100SO) or fish (100FO) oils as unique oil source. Intestinal fatty acid composition plus relative expression of intestinal EPs and other genes related with eicosanoid cascade were determined after 90 days. Moreover, at the end of the experiment a stress test (5 min of net chasing) was carried out. The use of VO altered the fatty acid profile of intestine and the expression of EP genes. The use of SO (rich in n-6 fatty acids) induced an overexpression of cox-2 in basal (non-stress) situation, plus a down-regulation of intestinal ep4 gene expression. Besides, fish fed soybean oil in the diet seems to be unable to respond properly to chasing stress.

A 12-week feeding experiment in indoor flowing seawater system was conducted to investigate the effects of graded levels of dietary fish protein hydrolysate (FPH) on growth performance and lipid accumulation of juvenile turbot (Scophthalmus maximus) (initial body weight 4.16 ± 0.01 g). Four isonitrogenous and isoenergetic experimental diets with high plant protein were formulated to contain graded levels of FPH, meanwhile the fish meal was replaced correspondingly by 0% (Diet FPH-0, control), 5% (Diet FPH-5), 10% (Diet FPH-10) and 20% (Diet FPH-20) of total dietary protein, respectively. Quadruplicate groups of 25 fish were fed to apparent satiation twice daily during the feeding trial. The results showed that the specific growth rate (SGR), protein efficiency ratio, and protein retention was not significantly different among group FPH-0, FPH-5, and FPH-10. The highest level of dietary FPH (FPH-20) significantly reduced the SGR but increased the feed intake compared to the control group. The viscerosomatic index in group FPH-10 and FPH-20 were significantly lower than that in the control group. Fish fed FPH-20 also showed significantly lower crude lipid concentration in whole body than fish fed the control diet. The concentrations of total and neutral lipid in gut were significantly lower in fish fed FPH-10 compared to the control group. For muscle lipid, the polar lipid concentration significantly decreased while the neutral lipid concentration significantly increased with increasing levels of dietary FPH, but no significant difference in total lipid concentration was observed among experimental groups. The lipid concentrations in liver were not significantly different among dietary treatments. With increasing levels of dietary FPH, serum triacylglycerol and cholesterol concentrations significantly decreased. The influence of dietary FPH on tissue fatty acid compositions generally corresponded with that on tissue lipid concentrations. In conclusion, these results suggested that in high plant protein diets FPH replacing fish meal by 10% of total dietary protein did not compromise the growth of juvenile turbot. However, a higher FPH level (replacing fish meal by 20% of total dietary protein) reduced the growth and feed utilization but it increased the feed intake. In the present experimental conditions, the FPH treatments, especially at high levels, significantly modulated the lipid accumulation and fatty acid compositions in turbot tissues, in a dose- and tissue-dependent manner.This study provided useful data for the circulatory use of fish processing by-products in fish diets. Also, based on the results of lipid analysis in fish tissues, potential lipid manipulating feed additives could be explored from fish protein hydrolysates.

The genetic mechanisms underlying host specificity of parasitic infections are largely unknown. After hatching, the larvae of the monogenean parasite, Heterobothrium okamotoi, attach to the gill filaments of hosts and the post-larval worms develop there by consuming nutrients from the host. The susceptibility to H. okamotoi infection differs markedly among fish species. While this parasite can grow on tiger pufferfish (also called fugu), Takifugu rubripes, it appears to be rejected by a close congener, grass pufferfish, Takifugu niphobles, after initial attachment to the gills. To determine the genetic architecture of the pufferfish responsible for this host specificity, we performed genome-wide quantitative trait loci analysis. We raised second generation (F2) hybrids of the two pufferfish species and experimentally infected them with the monogenean in vivo. To assess possible differences in host mechanisms between early and later periods of infection, we sampled fish three h and 21 days after exposure. Genome scanning of fish from the 3 h infection trial revealed suggestive quantitative trait loci on linkage groups 2 and 14, which affected the number of parasites on the gill. However, analysis of fish 21 days p.i. detected a significant quantitative trait locus on linkage group 9 and three other suggestive quantitative trait loci on linkage groups 7, 18 and 22. These results indicated the polygenic nature of the host mechanisms involved in the infection/rejection of H. okamotoi. Moreover the analyses suggested that host factors may play a more important role during the growth period of the parasite than during initial host recognition at the time of attachment. Within the 95% confidence interval of the linkage group 9 quantitative trait locus in the fugu genome, there were 214 annotated protein-coding genes, including immunity-related genes such as Irak4, Muc2 and Muc5ac.

Introduction: Neuroendocrine tumors are rare neoplasms that present bilateral liver metastases at diagnosis. Because of the good initial results obtained, liver transplantation is considered as a potentially curative treatment. The aim is to present our experience in liver transplantation in a series of 22 patients with unresectable bilateral hepatic metastases of neuroendocrine tumors and with previous resection of the primary tumor. Methods: We retrospectively reviewed the medical records of 22 patients (12 men and 10 women), with a mean age of 49,7 years, who underwent liver transplantation due to bilateral neuroendocrine liver metastases with previous resection of the primary tumour during 21 years. The most frequent location of the primary tumor was the pancreas in 13 patients (2 carcinoid, 3 gastrinoma, 1 glucagonoma and 8 non-functioning tumors). The remaining 9 tumors were located in the small bowel (7) and in the lung (2). Results: Only 1 patient died due to technical complications related to the transplant, representing a mortality rate for the entire group of 4,5 %. After a median follow-up of 11 years (range: 1 month-21 years), two patients died due to tumoral recurrence at 15 and 17 months. The survival rate at 3 and 5 years was 86% and 57%, respectively. Conclusions: In our series, the results the liver transplantation in the management of unresectable neuroendocrine liver metastases indicate that careful patient selection is required. The key to obtaining good results is individualization of the indication for this procedure.

The main objective of this study was to assess the effects of graded levels of dietary arachidonic acid (ARA), supplemented from alternative sources, on fatty acid composition of plasma and head kidney leucocytes of European sea bass (Dicentrarchus labrax). For that purpose, sea bass juveniles were fed four diets containing graded levels of ARA as follows: 0.5% (ARA0.5), 1% (ARA1), 2% (ARA2) and 4% (ARA4) during 60 days. At the end of the feeding trial fatty acid profiles of plasma and head kidney leucocytes were analyzed. Besides, plasma prostaglandins levels, head kidney leucocytes respiratory burst activity; peroxidase activity and phagocytic index were assayed. Reducing dietary ARA levels below 1% markedly reduced European sea bass growth performance. However, fish fed diet ARA0.5 tried to compensate this dietary ARA deficiency by a selective deposition of ARA on plasma and head kidney leucocytes, reaching similar levels to those fish fed diet ARA1 after 60 days of feeding. Nevertheless, head kidney phagocytic capacity was reduced as dietary ARA content in relation not only to variations on membrane composition but also to changes on fish basal prostaglandins levels. Results obtained demonstrated the importance to supply the necessary quantity n-6 LC-PUFA, and not only n-3 LC-PUFA levels, in European sea bass diets, in relation to not only growth performance but also immune system function.

Identifying membrane proteins and their multi-functional types is an indispensable yet challenging topic in proteomics and bioinformatics. In this article, we provide data that are used for training and testing Mem-ADSVM (Wan et al., 2016. “Mem-ADSVM: a two-layer multi-label predictor for identifying multi-functional types of membrane proteins” [1]), a two-layer multi-label predictor for predicting multi-functional types of membrane proteins.

We hypothesized that preemptive fibrinogen administration to obtain an initial plasma level of 2.9 g/L would reduce transfusion requirements in liver transplantation. A randomized, multicenter, hemoglobin-stratified, double-blind, fibrinogen-versus-saline-controlled trial was conducted. The primary end point was the percentage of patients requiring red blood cells. We evaluated 51 patients allocated to fibrinogen and 48 allocated to saline; the primary end point was assessed using data for 92 patients because the electronic record forms were offline for three patients in the fibrinogen group and four in the saline group. We injected a median of 3.54 g fibrinogen preemptively in the fibrinogen group. Nine patients in the saline group (20.9%) required fibrinogen at graft reperfusion (compared with one patient [2.1%] in the fibrinogen group; p = 0.005). Blood was transfused to 52.9% (95% confidence interval [CI] 42.5-63.3%) in the fibrinogen group and 42.74% (95% CI 28.3-57.2%) in the saline group (p = 0.217). Relative risk for blood transfusion was 0.80 (95% CI 0.57-1.13). Thrombotic events occurred in one patient (2.1%) and five patients (11.4%) in the fibrinogen and saline groups, respectively. Seven patients (14.6%) in the fibrinogen group and nine (20.3%) in the saline group required reoperation. Preemptive administration of fibrinogen concentrate did not influence transfusion requirements.

Streptococcus iniae is a Gram-positive bacteria that causes invasive infections with severe septicemia and meningitis, producing high economic losses in marine and continental aquaculture. Head kidney leukocytes of European sea bass (Dicentrarchus labrax) were used to measure the differential innate immune response upon infection with S. iniae. The complete inhibition in the production of intracellular superoxide radicals and total peroxidase content was observed in infected cells. This study also elucidates changes in the relative expression of some immune-related genes. Interleukin 1 beta, tumor necrosis factor-a and interleukin-6 reached a peak of expression at 4-8 h post-infection, subsequently decreasing significantly up to 48 h post-infection. However, interleukin-10 and Mx protein increased over time, reaching the pick of expression at 48 h post-infection, whereas caspase-3 showed down regulation until 48 h post-infection. The in vivo study of immune related genes show the same kinetics of mRNAs expression as in vitro experience. The proinflammatory cytokines mRNA transcription levels peaked at an earlier time in vivo than in vitro system. Our findings indicate that there is a direct relationship between the dissemination of bacteria and the resulting infection-associated inhibition of respiratory burst, apoptosis, and the pro- and anti-inflammatory gene expression profiles. (C) 2015 Published by Elsevier Ltd.