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Abstract Background Increasing demand for high-value fish species and pressure on forage fish is challenging aquaculture to ensure sustainable growth by replacing protein sources in aquafeeds with plant and terrestrial animal proteins, without compromising the economic value and quality of the final fish product. In the present study, the effects of a plant protein-based diet (CV), two plant-based diets in which graded amounts of plan protein mixtures were replaced with Hermetia illucens meal alone (VH10) or in combination with poultry by-product meal (PBM) (VH10P30), a fishmeal (FM) diet (CF) and an FM diet supplemented with H. illucens (FH10) on growth performance, gut health and homeostasis of farmed subadult European seabass were tested and compared. Results Fish fed the VH10 and VH10P30 diets showed the highest specific growth rates and lowest feed conversion ratios among the tested groups. Expectedly, the best preservation of PI morphology was observed in fish fed the CF or FH10 diets, while fish fed the CV diet exhibited significant degenerative changes in the proximal and distal intestines. However, PBM supplementation mitigated these effects and significantly improved all gut morphometric parameters in the VH10P30 group. Partial substitution of the plant mixture with insect meal alone or PBM also induced most BBM genes and activated BBM enzymes, suggesting a beneficial effect on intestinal digestive/absorption functions. Regarding intestinal microbiota, fish fed diets containing H. illucens meal (FH10, VH10, VH10P30) had the highest richness of bacterial communities and abundance of beneficial genera such as Lactobacillus and Bacillus. On the other hand, fish fed CV had the highest microbial diversity but lost a significant component of fish intestinal microbiota, the phylum Bacteroidetes. Finally, skin pigmentation most similar to that of farmed or even wild seabass was also observed in the fish groups fed CF, FH10 or VH10P30. Conclusion Plant-based diets supplemented with PBM and H. illucens pupae meal have great potential as alternative diets for European seabass, without affecting growth performance, gut homeostasis, or overall fitness. This also highlights the importance of animal proteins in diets of European seabass, as the addition of a small amount of these alternative animal protein sources significantly improved all measured parameters.

Abstract Background Brown trout is one of the most widespread fresh-water fish species in Europe. The evolutionary history of and phylogenetic relationships between brown trout populations are complex, and this is especially true for Italian populations, which are heavily influenced in different ways by stocking practices. The characterization of the genetic structure of Italian brown trout populations may give information on the risk of losing endemic Italian populations due to lack of genetic diversity or to admixture with stocking populations. The identification of signatures of selection, and the information deriving from dense genotyping data will help genotype-informed breeding programs. We used a ddRAD-seq approach to obtain more than 100,000 single nucleotide polymorphisms (SNPs), and to characterize the population structure and signatures of selection in 90 brown trout samples. Results Italian brown trout populations are genetically differentiated, although the stocking practices have introduced strong admixture in endemic Italian trout, especially with the Atlantic lineage. Most of the analysed populations showed high levels of kinship and inbreeding. We detected putative signatures of selection using different approaches, and investigated if the regions were enriched for functional categories. Several regions putatively under selection and characterized by a reduction in heterozygosity across all the studied populations are enriched for genes involved in the response to viral infections. Conclusions Our results, which show evidence of admixture with the Atlantic lineage (commonly used for stocking), confirm the need for controlling stocking practices, in order to avoid the erosion of the endemic gene pool; given the apparently high levels of kinship and inbreeding in local populations, our results also show the need to take action for increasing gene diversity. In addition, we used the genetically-distinct lineages to detect signatures of selection and we identified putative signatures of selection in several regions associated with resistance to infectious diseases. These constitute candidate regions for the study of resistance to infections in wild and farmed trout.

Abstract Background Proteinuria after kidney transplantation portends a worse graft survival. However the magnitude of proteinuria related to patient and graft survival and its correlation with donor and recipient characteristics are poorly explored. Methods This study investigated the impact of post transplant proteinuria in the first year in 1127 kidney transplants analyzing the impact of different donor ages. Proteinuria cut off was set at 0.5 g/day. Results Transplants with proteinuria > 0.5 g/day correlated with poor graft and patient outcome in all donor age groups. In addition, 6-month-1-year proteinuria increase was significantly associated with graft outcome, especially with donors > 60 years old (p 50 years old (Odd Ratio 2.3). Conclusions Post-transplant proteinuria was increasingly harmful with older donor age. Proteinuria ≥0.5 g/day correlates with worse outcomes in all transplanted patients. Prognostic value of proteinuria and eGFR for graft and patient survival was comparable and these two variables remain significant risk factors even in a multivariate model that take into consideration the most important clinical variables (donor age, rejection, delayed graft function and cytomegalovirus viremia among others).

This study compared the nutrient-energy retention, digestive function, growth performance, and welfare of rainbow trout (ibw 54 g) fed isoproteic (42%), isolipidic (24%), fishmeal-free diets (CV) over 13 weeks. The diets consisted of plant-protein replacement with graded levels (10, 30, 60%) of protein from poultry by-product (PBM) and black soldier fly H. illucens pupae (BSFM) meals, either singly or in combination. A fishmeal-based diet was also tested (CF). Nitrogen retention improved with moderate or high levels of dietary PBM and BSFM relative to CV (p < 0.05). Gut brush border enzyme activity was poorly affected by the diets. Gastric chitinase was up-regulated after high BSFM feeding (p < 0.05). The gut peptide and amino acid transport genes were differently regulated by protein source and level. Serum cortisol was unaffected, and the changes in metabolites stayed within the physiological range. High PBM and high BSFM lowered the leukocyte respiratory burst activity and increased the lysozyme activity compared to CV (p < 0.05). The BSFM and PBM both significantly changed the relative percentage of lymphocytes and monocytes (p < 0.05). In conclusion, moderate to high PBM and BSFM inclusions in fishmeal-free diets, either singly or in combination, improved gut function and nutrient retention, resulting in better growth performance and the good welfare of the rainbow trout.

Summary: We generated patient-specific disease-free induced pluripotent stem cells (iPSCs) from peripheral blood CD34+ cells and differentiated them into functional endothelial cells (ECs) secreting factor VIII (FVIII) for gene and cell therapy approaches to cure hemophilia A (HA), an X-linked bleeding disorder caused by F8 mutations. iPSCs were transduced with a lentiviral vector carrying FVIII transgene driven by an endothelial-specific promoter (VEC) and differentiated into bona fide ECs using an optimized protocol. FVIII-expressing ECs were intraportally transplanted in monocrotaline-conditioned non-obese diabetic (NOD) severe combined immune-deficient (scid)-IL2rγ null HA mice generating a chimeric liver with functional human ECs. Transplanted cells engrafted and proliferated in the liver along sinusoids, in the long term showed stable therapeutic FVIII activity (6%). These results demonstrate that the hemophilic phenotype can be rescued by transplantation of ECs derived from HA FVIII-corrected iPSCs, confirming the feasibility of cell-reprogramming strategy in patient-derived cells as an approach for HA gene and cell therapy. : Follenzi and colleagues show that endothelial cells can be obtained starting from blood CD34+ cells of both healthy and hemophilic donors passing through iPSC generation. During the differentiation process the cells can be genetically modified by LV to express FVIII; corrected endothelial cells secrete FVIII in vitro and ameliorate the hemophilic phenotype in a mouse model of the disease. Keywords: hemophilia A, induced pluripotent stem cells (iPSCs), endothelial cells, cell and gene therapy, FVIII, chimeric vasculature, lentiviral vectors

Salmonid fish have become ecological and research models of study in the field of conservation genetics and genomics. Over the last decade, brown trout have received a high level of interest in research and publications. The term Salmo trutta complex is used to indicate the large number of geographic forms present in the species Salmo trutta. In Europe, the S. trutta complex consists (based on mitochondrial DNA control region analysis) of seven major evolutionary lineages: Atlantic (AT), Mediterranean (ME), Adriatic (AD), Danubian (DA), Marmoratus (MA), Duero (DU) and Tigris (TI). In several nations, the difficulty of identifying some lineages derives from their wide phenotypic and geographic plasticity and the presence of mixed lineages (due to introgressive hybridization with domestic AT populations). In Italy, the S. trutta complex populations living in the Tyrrhenian area and on the main islands (Sicily, Sardinia and Corsica) showed high genetic diversity. Currently, on the Italian Red List, the protected (near threatened) populations are the AD and ME lineages. Recent studies based on traditional (mitochondrial and nuclear markers) and NGS (next-generation sequencing) analyses have clarified some genetic differences between the populations of the Tyrrhenian region, Sicily, Sardinia and Corsica. Native populations in Sardinia belong to the AD lineage, while those living in Corsica are mainly characterized by the AD, MA and ME haplotypes. In Sicily, in the area of the Iblei mountains, an AT lineage (North African) exists. According to some authors, the term Salmo macrostigma should only be used for populations in North Africa. The use of genotyping methods based on mtDNA and nuclear markers and the latest generation sequencing techniques can improve the study of populations and evolutionary lineages in areas where there are overlaps and hybridization phenomena.

Background: Central Venous Catheter (CVC)-related infections cannot always be solved by replacement, due to some vascular anomalies or an emergency status. This comprehensive, evidence-based review aimed to define the efficacy of antibacterial lock therapy (ALT) compared to the standard of care (SoC) in CVC-related infections. Methods: We performed a systematic search in PubMed, Embase, and Google Scholar, looking for randomized controlled trials (RCTs) and cohort or case-control observational studies. The eligible studies considered the subjects with a diagnosis of CVC-related infections treated with antibacterial lock therapy (ALT) compared to the standard of care (SoC). Results: Among 609 records at the end of the selection process, five articles, referring to observational studies, were included in this systematic review. In pooled analyses, including a total of 276 individuals, microbiological healing (OR 3.78; 95% CI; 2.03–7.03) showed significant differences between ALT and the SoC, with a follow-up varying from 2 weeks to 3 months. Conclusions: Our results suggested that ALT could improve the preservation of CVCs and could be considered when their replacement is not possible as a result of vascular problems. However, only observational studies were included and RCTs are needed to confirm these findings and to increase the level of evidence.

Polyomavirus-associated nephropathy is an important cause of allograft dysfunction and graft loss after kidney transplantation. Even if histological evaluation is the gold standard for graft study and diagnosis of polyomavirus-associated nephropathy, K-DIGO guidelines suggest performing an indication biopsy in selected patient’s clinical conditions or laboratory parameters. The practice of protocol biopsy is still controversial. We report the management of a case of presumptive polyomavirus-associated nephropathy in a 53-year-old kidney transplant recipient affected by type 1 hyperoxaluria with persistent high levels of viruria and sustained levels of polyomavirus BK viremia. The presence of a presumptive polyomavirus-associated nephropathy, even if never confirmed by biopsy, never compromised his clinical condition and allograft function. As a result of an immunosuppression-sparing policy and use of mTOR inhibitor, the polyomavirus BK viremia was successfully controlled with an observation time >5 years. The decision to perform or not a graft biopsy was the main question in the management of this case. We opted for a non-invasive approach because of the high risk of biopsy with macrohematuria on earlier biopsy in a dual kidney transplant and patient’s unwillingness for the procedure. The replication level of polyomavirus BK was significantly reduced by the decrease of immunosuppression on the basis of a close nucleic acid testing monitoring. The strategy we adopted could be considered in cases when renal biopsy is contraindicated or considered to be high risk.

This study investigated the effects of dietary inclusion levels of full-fat Hermetia illucens prepupae meal (H) on growth and gastrointestinal integrity in rainbow trout (Oncorhynchus mykiss). A 98-day study was conducted using triplicate groups of trout (initial body weight, 137 ± 10.5 g) kept in 1-m3 tanks in a flow-through well water system. Three dietary treatments were prepared: one based on fishmeal and purified protein-rich vegetable ingredients (H0), and two experimental diets including graded levels of H meal (25% and 50%, referred to as H25 and H50, respectively). At the end of the feeding trial, no differences were observed in growth performance and plasma metabolite levels, with the biometric data confirmed by the liver expression of the genes involved in somatic growth regulation (igf1 and mstn1a). In the H50 group, a three-fold up regulation of liver hsp70 was observed. An activation of the stress/immune response (il-10, tnf-α, and tlr-5) was observed in medium intestine in the H25 and H50 groups (p < 0.05) together with a villi length reduction detected through histological analyses. Liver histology and Fourier Transform Infrared Imaging (FTIRI) spectroscopy highlighted an increase in lipid deposition. These findings suggest that caution should be taken into account when 50% replacement of conventional ingredients with H is selected.

Delayed graft function (DGF) is an early complication of kidney transplantation (KT) associated with increased risk of early loss of graft function. DGF increases using kidneys from extended criteria donors (ECD). NGAL is a 25KDa protein proposed as biomarker of acute kidney injury. The aim of this study was to investigate the role of NGAL as an early and accurate indicator of DGF and Tacrolimus (Tac) toxicity and as a mediator of tissue regeneration in KT from ECD.We evaluated plasma levels of NGAL in 50 KT patients from ECD in the first 4 days after surgery or after Tac introduction.Plasma levels of NGAL at day 1 were significantly higher in DGF group. In the non DGF group, NGAL discriminated between slow or immediate graft function and decreased more rapidly than serum creatinine. NGAL increased after Tac introduction, suggesting a role as marker of drug toxicity. In vitro, hypoxia and Tac induced NGAL release from tubular epithelial cells (TEC) favoring an autocrine loop that sustains proliferation and inhibits apoptosis (decrease of caspases and Bax/Bcl-2 ratio).NGAL is an early and accurate biomarker of graft function in KT from ECD favoring TEC regeneration after ischemic and nephrotoxic injury.

The American lobster (Homarus americanus) is one of the most important crustacean resources in North America. In Italy and Europe, this fishery product is available throughout the year and it has a high and increasing commercial demand. American lobsters are traditionally marketed live and stocked, without feed, in temperature controlled recirculating systems for several weeks before being sold in the market places. The current Italian legislation does not fix a maximum length of time for the crustacean confinement and specific welfare requirements. In the present research, a 4-week experiment was carried out using 42 adult H. americanus reared in 4 recirculating aquaculture tanks. After one month of confinement, mean glucose, protein and total haemocyte count levels in the hemolymph of H. americanus were stable and similar (P>0.05) to the values observed at the beginning of the experiment. Results of the proximate analysis of the abdominal muscles of H. americanus showed no significant differences in concentrations of crude protein, lipid and ash during the trial. At the end of the experiment, the sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting analysis revealed a marked degradation of the muscle myofibrillar proteins. A number of fragments, possibly from myosin, were evident in the range between 50 and 220 kDa between time t0 and t28. Results of this study show that the main hemolymphatic variables and degradation analysis of the muscle myofibrillar proteins can be used as sensitive indicators of the crustacean stress response to confinement and starvation.

The effects of feeding two complete extruded diets differing in protein source (fish meal-FM vs. vegetable proteins-VP) over 30 weeks and subsequent 30 days of starvation on biometry, fillet composition and liver IGF-I mRNA were studied in large rainbow trout. At the end of the feeding period, the dietary protein source little affected major biometry traits, dressing out yields and overall adiposity (P>0.05) but fish given the VP diet resulted in higher content of PUFA n-6 fatty acids in mus- cle (0.46 vs. 0.22 g/100g fillet, P0.05) and of all fatty acids in fillet (P

The effect of the farming system on biometry traits and dressing out yield were inves- tigated in market-size European sea bass (Dicentrarchus labrax) cultured extensively or intensively in sea cages or land-based basins. Fish external appearences and shapes were studies with geometric morphometrics in order to assess the potential of combined methodologies in the assessment of finfish quality. Both standard biometry and geometric morphometrics were able to discriminate between sea bass farmed extensively from those cultured under intensive conditions. Geometric morphometrics has been shown to be a valuable tool for describing changes in shape features and could result a useful technique to be associated to biometry traits in the context of fish quality assessment.

The aim of this trial was to study the effect of supplementary feeding offered to farmed fallow deer does on milk characteristics and on male and female fawns performance. During two years, 40 females were divided in two groups and assigned to two homogeneous paddocks of the same pasture: SUP group received 0.5 kg/d per female of supplement that was raised to 0.7 kg/d from the third month of fawns age, while HER group was fed only on herbage. Herbage availability and composition were monitored by exclusion cages. Female performance (body weight, body condition score and milk composition) was recorded. Further, during the first year, performance and hematological parameters (non-esterified fatty acids and urea nitrogen) were assessed. Fawns were weighed every 45 days until the end of the experiment, when the characteristics of their carcasses were recorded. Supplementary feeding reduced herbage gathering and quality, but allowed a full recovery of body weight and condition of does. The milk provided by the HER does to their female offspring was richer in fat (16.1 vs 14.2 g/100 mL) and protein (8.76 vs 8.04 g/100 mL) than that furnished to male fawns; the opposite condition happened for SUP does (12.6 vs 13.3 g/100 mL for fat and 8.04 vs 8.35 g/100 mL for protein). At slaughter, SUP fawns showed higher carcass weight (14.10 vs 11.26 kg), greater conformation score (2.1 vs 1.6 points) and higher fat score (2.0 vs 1.2 points) than HER ones, moreover male fawns presented higher carcass weight than female (13.62 vs 11.74 kg).

Background. A pathogenic role for polyomavirus BK in systemic lupus erythematosus (SLE) has been proposed, however no study evaluated the occurrence of BK replication in renal transplant recipients according to the underlying disease leading to transplantation and its potential impact. Methods. The occurrence of BK reactivation was serially evaluated in 468 renal transplant recipients, including 11 patients with SLE as underlying disease (overall, 2370 serum and 2370 urine specimens; 65 from SLE patients). Results. Considering the overall occurrence of viral reactivation (viremia and/or viruria), 26/65 (40%) specimens were positive in four SLE patients (36.3%) versus 331/2143 (15.4%) in 130/227 (57.3%) non-SLE patients. A patient transplanted for class III lupus nephritis evidenced sustained BK viremia and viruria (with viremia values potentially indicative of polyomavirus-associated nephropathy) in the absence of clinical features of renal dysfunction or recurrence of lupus nephritis. Conclusions. Further studies on larger populations and for a longer follow-up should be required to evaluate the impact of BKV reactivation in renal transplant patients with SLE as underlying disease, as well as the potential therapeutic implications.

Introduction. Reactivation of polyomavirus BK in kidney transplant recipients has been associated to the development of nephropathy (polyomavirus-associated nephropathy, PVAN), possibly leading to the loss of the transplanted organ. Immunosuppression is the condicio sine qua non for the onset of PVAN; however, a lower incidence of BK viremia has been reported with low-level tacrolimus based immunosuppressive protocols in comparison to cyclosporine A.Aim of this study was to compare the two immunosuppressive protocols. Methods. Virological monitoring of BK was performed in 468 consecutive renal transplant patients over a period of 3 years (2370 urine e 2370 serum specimens): in particular, 1780 specimens from 362 patients treated with tacrolimus and 590 from 106 treated with cyclosporine A. Results. BK viremia was evidenced in 124 (7.0%) and 12 (2.0%) specimens from 40 (11.0%) and 11 (10.4%) patients treated with tacrolimus and cyclosporine A, respectively; similarly, BK viruria in 289 (16.2%) and 58 (9.8%) specimens from 67 (18.5%) and 27 (25.5%) patients, being the difference of incidence highly significant (p

Introduction. Human polyomavirus BK (BKV) is worldwide distributed, with a seroprevalence rate of 70–90% in the adults. Following primary infection, BK remains latent in the renourinary tract as the epidemiologically most relevant latency site, and in B cell, brain, spleen and probably other tissues. Reactivation may occur in both immunocompetent subjects and immunocompromised patients. In renal transplantation, in the context of intense immunosuppression, viral replication may determine BKV-associated nephropathy (BKVAN) with interstitial nephritis and/or ureteral stenosis in 1–10% of the patients and leading to graft failure and return to haemodialysis in 30 to 80% of the cases (5). Screening of BKV replication represents the basic strategy to predict early the onset of BKVAN and may allow for earlier intervention with reduced allograft loss (3, 4). Nowadays, replication of BKV is monitored by quantification of BKV-DNA in serum and urine (2). The aim of this study was to evaluated the role of BKV VP1 mRNA in urine as a marker of viral replication in renal transplant recipients.

The black soldier fly (BSF) (Hermetia illucens L.) is one of the most promising species for the production of ingredients, mainly protein, useful for animal feed formulation, owing to its ability to convert organic waste into biomass with a high nutritional value. However, the low percentage of n-3 series polyunsaturated fatty acids (PUFAs) in its fatty acid profile is a limiting factor for the utilization of BSF in fish feed. Recent studies have highlighted that wastes derived from different agro-food value chains could affect the nutritional composition of BSF larvae, depending on the composition of the wastes used as larvae-growing substrate. Due to the significant amount of n-3 PUFA in marine products, both in edible produce and in by-products, in this study, different sources of fish by-products were included in experimental diets for BSF to evaluate the effect of its addition on the final nutritional profile of BSF pre-pupae. One control diet and five experimental diets were prepared to feed the BSF larvae: wheat bran as the control diet (diet B), bycatch from Mediterranean trawl fisheries (diet F), Parapenaeus longirostris processing by-products (diet S), aquaculture processing by-products (diet R), Thunnus albacares processing by-products (diet T), and Engraulis encrasicolus processing by-products (diet A). In this study, the effects of the different diets were analyzed on the growth, body composition, and fatty acid profile of BSF larvae and pre-pupae. The obtained results showed that the different experimental diets affected total lipids content and fatty acids composition, when compared with the control. A significant increase in eicosapentaenoic (EPA) and docosahexaenoic acid (DHA) in BSF larvae and pre-pupae fed with all fish by-products was observed when compared with those fed with diet B, in particular in larvae and pre-pupae fed with diet A, demonstrating that the utilization of fish processing by-products is a suitable solution for improving the nutritional value of insects as ingredients in aqua feeds. The reuse of marine by-products can contribute to the industry’s “zero waste” goal, increasing the sustainability of the fishery value chain and the formulation of new valuable products.

The use of natural immunostimulants is considered the most promising alternative to promote fish health, productive performance and quality, increasing the aquaculture profitability, sustainability and social acceptance. The purpose of this study was to evaluate the effect of the integration of a potential probiotic strain, Halobacterium salinarum, belonging to the Archaea domain, in the formulated diets of farmed gilthead seabream (Sparus aurata L.) in terms of growth performances and immunity responses. The experiment was set up to test two different levels of inclusion of the bacteria in the diet: 0.05% (D1) and 0.1% (D2). The effects on fish growth performances; humoral (peroxidase, protease, antiprotease and IgM levels) and cellular immunity parameters (phagocytosis, respiratory burst and myeloperoxidase), along with bactericidal activity, were evaluated after 15 and 30 days of experimental feeding. The obtained results showed that the inclusion of H. salinarum at the highest concentration (D2 0.1%) improved growth performances, bactericidal activity against Vibrio anguillarum and some parameters related both to the humoral and cellular immune response, suggesting exploring other aspects of welfare in view of future supplementations of this probiotic strain in the diet of S. aurata.

Rosmarinus officinalis L. is a dietary source that produces polyphenols as secondary metabolites. These natural compounds with potent antioxidant abilities are increasingly recommended as a supplement to inhibit oxidative stress. In the current work, we evaluated the impact of the extraction method on the chemical composition of R. officinalis extract, especially on the content of carnosic (CA) and rosmarinic (RA) acids using UPLC-MS-DAD as well as on their antioxidant potency. Four extracts of Tunisian rosemary were obtained from non-conventional extraction techniques:ultrasound-assisted extraction (UAE),supercritical extraction (SFE) and UAE and SFE combined ((UAE-SFE(I), UAE-SFE(II)). The UAE exhibited the best total phenolic compounds (i.e., 85.27 mg GAEg−1), the highest content of CAand RA and the strongest antioxidant abilities (i.e., IC50 = 0.13 mg/mL and EC50 = 0.93 mg/mL for DPPH scavenging test and iron reducing power ability assay). The evaluation of antioxidant activity of UAE inhuman skin fibroblast (HS-68) cell line was carried out after the induction of oxidative stress. The results determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay showed a strong protective effect against H2O2oxidative stress induction in cells pretreated with UAE. The obtained results allow us to give new insight about the effect of the extraction method on the chemical composition and biological activities of the extract and the importance of the choice of the most appropriate processing technique to prepare rosemary extract with a high antioxidant potency and protective effect against oxidative stress.