Effects of the feeding system on performance and myopathy occurrence in two broiler chicken genotypes

To evaluate whether performance and myopathy occurrence differed according to genotype (Cobb 500 vs. Ross 308) and feeding system (AL: ad libitum vs. ER: early restricted from 13 to 23 d of age, vs. LR: late restricted from 27 to 37 d; restriction rate: 80% of ad libitum), 828 day- old male chicks were assigned to 6 groups (2 x 3 arrangement), housed in 36 pens, and controlled for: individual live weight (weekly) and pen feed intake (daily) until slaughter (48 d); white striping and wooden breast occurrence at slaughter. Individual data were analysed by PROC MIXED of SAS (fixed effects: feeding system, genotype, and interaction; random effect: pen); pen feed intake and myopathy occurrence were analysed by PROC GLM and CATMOD, respectively. The feeding system affected performance: at the end of the first period (1-22 d), ER chickens showed lower weight gain (40.5 g/d vs. 47.8 g/d and 48.0 g/d), feed intake (50.8 g/d vs 61.0 g/d and 60.2 g/d) and weight (903 g vs 1056 g and 1059 g) than AL and LR broilers (P<0.001); at the end of the second period (23-48 d), ER chickens showed higher weight gain (98.3 g/d vs 93.6 g/d and 90.2 g/d) and feed intake (182 g/d vs. 177 g/d and 171 g/d) compared to AL and LR chickens (P<0.001). Final live weight was the highest in AL group, intermediate in ER group, and the lowest in the LR one (3482 g, 3454 g, and 3399 g; P<0.01). Feed conversion in the whole period did not change with the feeding system. At gross examination, white striping occurrence changed from 77.8% to 67.1%, and 81.7% in AL, ER and LR broilers (p<0.10). Differences between genotypes were evident from the first day and, at the end of the trial, weight gain (74.3 g/d vs 70.1 g/d), feed intake (126 g/d vs 114 g/d), feed conversion (1.69 vs 1.64), and live weight (3548 g vs. 3342 g) were higher in the Ross than in the Cobb chickens (P<0.001). At slaughter, the rate of white-striped breasts was similar (on average 75.5%), but the occurrence of severely white-striped breasts was higher in the Ross than in the Cobb chickens (25.9% vs. 7.41%; P<0.001). Wooden breast occurrence (on average 5.1%) did not change with the feeding system or the genotype. In conclusion, under our conditions, a late feed restriction did not permit to recover performance at the end of the trial nor to control white striping occurrence. Moreover, the genotype affected growth rate and white striping degree: the highest the growth rate, the highest the severity of white striping

Characterization of the human D-amino acid oxidase (hDAAO) - pLG72 complex involved in the onset of schizophrenia.

Schizophrenia is a chronic and severely debilitating psychiatric disorder affecting nearly 1% of the world’s population. In 2002, the new human gene G72, encoding for the pLG72 protein, and the gene encoding for DAAO have been genetically linked to the susceptibility to schizophrenia. A yeast two-hybrid screening experiment identified D-amino acid oxidase (DAAO) as a putative interacting partner of pLG72. DAAO is a FAD-containing flavooxidase that in brain is responsible for the elimination of D-serine, a co-agonist that binds to the glycine-site of the NMDA receptor. We recently demonstrated that pLG72 acts as “inactivator” of human DAAO and that the cellular concentration of D-serine depends on the expression of the active form of this flavooxidase. Based on these results, a molecular model for the onset of schizophrenia has been proposed: a decrease in pLG72 expression might yield an anomalous high level of hDAAO activity and therefore a decrease in the local concentration of D-serine, affecting glutamatergic neurotransmission mediated by NMDA receptor. The characterization of the complex is a challenging task, hardly feasible by high resolution techniques, since: 1) structural informations on pLG72 are lacking; 2) pLG72 is soluble only in the presence of mild denaturant; 3) no homologous protein has been structurally characterized so far. In this perspective, we have used low resolution strategies based on the coupling of classical biochemistry approaches (complementary proteolysis, cross-link) with mass spectrometric techniques, to characterize the pLG72-hDAAO complex. Results indicated that hDAAO exhibits different proteolysis profiles when isolated or in complex with pLG72, thus suggesting a conformational change upon binding the effector protein. Chemical cross-linking experiments will complement the proteolysis experiments providing with details the contact regions between hDAAO e pLG72

Structural characterization of the M* partly folded intermediate of wild-type and P138A EcAspAT

A combination of spectroscopic techniques, hydrogen/deuterium exchange, and limited proteolysis experiments coupled to mass spectrometry analysis was used to depict the topology of the monomeric M*partly folded intermediate of aspartate aminotransferase from Escherichia coli in wild type (WT) as well as in a mutant form in which the highly conserved cis-proline at position 138 was replaced by a trans-alanine (P138A). Fluorescence analysis indicates that, although M* is an off-pathway intermediate in the folding of WT aspartate aminotransferase from E. coli, it seems to coincide with an on-pathway folding intermediate for the P138A mutant. Spectroscopic data, hydrogen/deuterium exchange, and limited proteolysis experiments demonstrated the occurrence of conformational differences between the two M*intermediates, with P138A-M* being conceivably more compact than WT-M*. Limited proteolysis data suggested that these conformational differences might be related to a different relative orientation of the small and large domains of the protein induced by the presence of the cis-proline residue at position 138. These differences between the two M* species indicated that in WT-M* Pro138 is in the cis conformation at this stage of the folding process. Moreover, hydrogen/deuterium exchange results showed the occurrence of few differences in the native N2 forms of WT and P138A, the spectroscopic features and crystallographic structures of which are almost superimposabl

PHK from phenol hydroxylase of Pseudomonas sp. OX1. Insight into the role of an accessory protein in bacterial multicomponent monooxygenases

Bacterial multicomponent monooxygenases (BMMs) are members of a wide family of diiron enzymes that use molecular oxygen to hydroxylate a variety of aromatic compounds. The presence of genes encoding for accessory proteins not involved in catalysis and whose role is still elusive, is a common feature of the gene clusters of several BMMs, including phenol hydroxylases and several soluble methane monooxygenases. In this study we have expressed, purified, and partially characterized the accessory component PHK of the phenol hydroxylase from Pseudomonas sp. OX1, a bacterium able to degrade several aromatic compounds. The phenol hydroxylase (ph) gene cluster was expressed in Escherichia coli/JM109 cells in the absence and in the presence of the phk gene. The presence of the phk gene lead to an increase in the hydroxylase activity of whole recombinant cells with phenol. PHK was assessed for its ability to interact with the active hydroxylase complex. Our results show that PHK is neither involved in the catalytic activity of the phenol hydroxylase complex nor required for the assembly of apo-hydroxylase. Our results suggest instead that this component may be responsible for enhancing iron incorporation into the active site of the apo-hydroxylase

Effect of dietary supplementation with insect fats on growth performance, digestive efficiency and health of rabbits

Background: The present work aimed at evaluating the effect of the dietary replacement of soybean oil (S) by two types of insect fats extracted from black soldier fly larvae (H, Hermetia illucens L.) and yellow mealworm larvae (T, Tenebrio molitor L.) on growth performance, nutrient digestibility, blood parameters, intestinal morphology and health of growing rabbits. Methods: At weaning, 200 crossbred rabbits (36 days old) were allotted to five dietary treatments (40 rabbits/group): a control diet (C) containing 1.5% of soybean oil and four experimental diets where soybean oil was partially (50%) or totally (100%) substituted by H (H50 and H100) or T (T50 and T100) fats. Total tract digestibility was evaluated on 12 rabbits per treatment. The growth trial lasted 41 d and, at slaughtering (78 days old), blood samples were collected from 15 rabbits per treatment, morphometric analyses were performed on duodenum, jejunum and ileum mucosa, and samples of liver, spleen and kidney were submitted to histological evaluation. Results: No difference was observed between the control and the experimental groups fed insect fats in terms of performance, morbidity, mortality and blood variables. The addition of H and T fats did not influence apparent digestibility coefficients of dry matter, protein, ether extract, fibre fractions and gross energy. Gut morphometric indices and organ histopathology were not affected by dietary inclusion of H and T fats. Conclusions: H and T fats are suitable sources of lipid in rabbit diets to replace soybean oil without any detrimental effect on growth performance, apparent digestibility, gut mucosa traits and health

Limitations and challenges in protein stability prediction upon genome variations: towards future applications in precision medicine

Protein stability predictions are becoming essential in medicine to develop novel immunotherapeutic agents and for drug discovery. Despite the large number of computational approaches for predicting the protein stability upon mutation, there are still critical unsolved problems: 1) the limited number of thermodynamic measurements for proteins provided by current databases; 2) the large intrinsic variability of \u394\u394G values due to different experimental conditions; 3) biases in the development of predictive methods caused by ignoring the anti-symmetry of \u394\u394G values between mutant and native protein forms; 4) over-optimistic prediction performance, due to sequence similarity between proteins used in training and test datasets. Here, we review these issues, highlighting new challenges required to improve current tools and to achieve more reliable predictions. In addition, we provide a perspective of how these methods will be beneficial for designing novel precision medicine approaches for several genetic disorders caused by mutations, such as cancer and neurodegenerative diseases

Evaluation of carcass and meat traits of Muscovy duck fed with black soldier fly partially defatted meal

The aim of this study was to evaluate the carcass characteristics and breast meat quality in Muscovy duck (Cairina moschata domestica) fed different inclusion levels of a partially defatted black soldier fly larva (BSF) meal. A total of 256 Muscovy ducklings (average live weight, LW: 71.32\ub12.70 g) were reared from day 3 to day 48 and randomly allotted in 32 pens (8 replicates/treatment). Four different diets were formulated with increasing substitution level of corn gluten meal with BSF larva meal (0, 3, 6 and 9%; BSF0, BSF3, BSF6 and BSF9, respectively) and divided in 3 feeding phases: starter (1-14 days), grower (14-35 days) and finisher (35-48 days). At day 48, 2 animals/replicate were slaughtered and dissected to determine their carcass yields. The weights of spleen, bursa of Fabricius, liver, heart and abdominal fat were recorded. Breast and thigh muscles were then excised from 16 ducks/treatment and weighted. Ultimate pH (pHu) and L*, a*, b* colour values were then measured on breast muscle. The collected data were tested by means of oneway ANOVA evaluating the effect of dietary BSF inclusion level by polynomial contrasts. Significance was declared at P<0.05. The inclusion of BSF did not affect final LW (2,515.68\ub192.42 g on average). Hot and cold carcass weights showed a quadratic response (P<0.05) to increasing BSF larva meal, with a minimum corresponding to BSF6; however, refrigeration losses were not affected by treatments. Weight of spleen, bursa of Fabricius, liver and heart did not differ among treatments. The weight of abdominal fat showed a quadratic response to increasing BSF meal with a minimum corresponding to BSF6 group (P<0.05). Breast and thigh yields, pHu and L*, a*, b* colour values did not differ among groups. With the exception of BSF6, the inclusion of BSF meal did not affect meat traits and carcass characteristics, confirming the potential use of BSF meal in Muscovy duck diets

Crossbreeding to improve local chicken breeds: predicting growth performance of the crosses using the Gompertz model and estimated heterosis

The present study aims to validate the Gompertz model to predict the growth performance of chicken crosses according to growth curve parameters of the parental lines and the estimated heterosis for each curve parameter. A total of 252 one-day-old chicks of both sexes belonging to 6 genotypes, including Ross 308, Sasso (SA), Bionda Piemontese (BP), and Robusta Maculata (RM), and the crosses between these local breeds and SA (BP x SA and RM x SA) were randomly allocated in 18 pens (3 pens/genotype) in mixed-sex groups (14 animals/pen; 7 females and 7 males). The individual body weight (BW) of all birds was recorded once a week from hatching until slaughtering (81 d for Ross 308; 112 d for SA, 140 d for the other genotypes). We drew up our final dataset with 240 birds (40 birds/ genotype; 20 females and 20 males). The growth curve of each genotype was described using the Gompertz model, and the heterosis for each growth curve parameter was calculated as the difference between F-1 crosses and the average of parental breeds. The predicted growth curve parameters were evaluated by cross-validation. The Gompertz model accurately estimated the growth curves of all the genotypes (R-2 > 0.90). Heterosis was significant for almost all growth curve parameters in both crosses (P < 0.05). Heterosis ranged from -13.0 to +11.5%, depending on parameters, but varied slightly between the crossbreeds (BP x SA and RM x SA). The predicted values of adult BW, weight at the inflection point, and maximum growth rate were overestimated for BP x SA and under-estimated for RM x SA, with a mean error between observed and predicted values <12.71% for all the curve parameters. In conclusion, the growth performance of chicken crosses between local breeds and commercial strains can be accurately predicted with Gompertz parameters of the parental lines adjusting for heterosis

Quality and consumer acceptance of meat from rabbits fed diets in which soybean oil is replaced with black soldier fly and yellow mealworm fats

This trial investigated the effect of the dietary inclusion of Hermetia illucens (H) and Tenebrio molitor (T) fats as alternative lipid sources for growing rabbits, and assessed the carcass characteristics, proximate composition, lipid peroxidation, and fatty acid profile of the meat, as well as consumer acceptance. At weaning, 200 crossbred rabbits (1051\ub1138 g initial body weight) were allotted to five isolipidic (4% dry matter (DM)) dietary treatments: a control diet (C) containing 1.5% of soybean oil, and four experimental diets in which soybean oil was partially (50%) or totally (100%) substituted by H (H50 and H100) or T (T50 and T100) fats. The carcass characteristics, the meat quality traits, and the consumer acceptance of the cooked meat were not affected. The fat content of Longissimus thoracis et lumborum muscle of the rabbits was 1.1% on average. In the case of rabbit fed the H diets (average of diets H50 and H100), the same muscles revealed a higher saturated fatty acid proportion (47.1% vs. 39.7% and 40.8%, respectively) and a lower polyunsaturated fatty acid proportion than the rabbits fed the C and T diets (average of diets T50 and T100) (26.5% vs. 31.7% and 29.7%) (p<0.001). The meat of the rabbits fed the diets containing insect fat (average for H and T diets) was less susceptible to oxidation (0.24 vs. 0.39 mg malondialdehyde/kg meat in the C group; p<0.01)