Influence of dietary Chlorella vulgaris and carbohydrate-active enzymes on growth performance, meat quality and lipid composition of broiler chickens.

Herein, we investigated the effect of Chlorella vulgaris as ingredient (10% of incorporation) in broiler diets, supplemented or not with 2 formulations of Carbohydrate-Active enZymes (CAZymes; Rovabio Excel AP and a mixture of recombinant CAZymes, composed by an exo-ß-glucosaminidase, an alginate lyase, a peptidoglycan N-acetylmuramic acid deacetylase and a lysozyme), on growth performance, meat quality, fatty acid composition, oxidative stability, and sensory traits. One hundred twenty 1-day-old Ross 308 male birds were randomly assigned to one of the 4 experimental diets (n = 30): corn-soybean meal-basal diet (control), basal diet with 10% C. vulgaris (CV), CV supplemented with 0.005% of a commercial CAZyme cocktail (Rovabio Excel AP), (CV + R), and CV supplemented with 0.01% of a 4-CAZyme mixture previously selected (CV + M) during the experimental period lasted from day 21 to day 35. Body weight gain and feed conversion rate of broilers were not affected by C. vulgaris but digesta viscosity increased more than 2-fold (P < 0.001) relative to the control. In addition, neither cooking loss, shear force, juiciness, flavor nor off-flavor was impaired by dietary treatments (P > 0.05). By contrast, the dietary C. vulgaris increased tenderness, yellowness (b∗) and total carotenoids in breast and thigh meats. However, no additional protective effect against lipid oxidation was observed in meat with the inclusion of microalga. Chlorella vulgaris, independently of CAZymes, had a minor impact on meat fatty acid composition but improved the proportion of some beneficial fatty acids. In summary, our data indicate a slight improvement of broiler meat quality and lipid nutritional value, without impairment of broilers' growth performance, thus supporting the usefulness of this microalga in poultry diets, up to this high level of incorporation. By contrast, the selected CAZyme mixtures used do not significantly improve the release of microalga nutrients in poultry diets, through the disruption of microalga cell wall, which warrants further research.

The effects of improving low dietary protein utilization on the proteome of lamb tissues.

Cistus ladanifer L. is a common shrub endemic to the Mediterranean region with high levels of condensed tannins (CT). CT form complexes with dietary protein resisting microbial degradation in the rumen, which enhances dietary protein utilization in ruminant diets. The objective of this study was to evaluate the utilization of CT in the diet of lambs on the proteomes of muscle, hepatic and adipose tissues. Twenty-four Merino Branco ram lambs were randomly allocated to three treatments (n = 8): C - control (160 g crude protein (CP)) per kg DM, RP - reduced protein (120 g CP/kg DM); and RPCT - reduced protein (120 g CP/kg DM) treated with CT extract. At the end of the trial, lambs were slaughtered and the longissimus lumborum muscle, hepatic and peri-renal adipose tissues sampled. A two-way approach was used for proteomic analysis: 2D-DIGE and nanoLC-MS. In the muscle, C lambs had lower abundance proteins that partake in the glycolysis pathway than the lambs of other treatments. Control lambs had lower abundance of Fe-carrying proteins in the hepatic tissue than RP and RPCT lambs. The latter lambs had highest abundance of hepatic flavin reductase. In the adipose tissue, C lambs had lowest abundance of fatty-acid synthase. SIGNIFICANCE: soybean meal is an expensive feedstuff in which intensive animal production systems heavily rely on. It is a source of protein extensively degraded in the rumen, leading to efficiency losses on dietary protein utilization during digestion. Protection of dietary protein from extensive ruminal degradation throughout the use of plants or extracts rich in CT allow an increase in the digestive utilization of feed proteins. In addition to enhance the protein digestive utilization, dietary CT may induce other beneficial effects in ruminants such as the improvement of the antioxidant status.

The effect of Nannochloropsis oceanica feed inclusion on rabbit muscle proteome.

Nannochloropsis oceanica is a microalga, highly concentrated in protein and EPA (eicosapentaenoic acid). It has a recalcitrant cell wall that decreases nutrient digestibility. The objective of this study is to evaluate the effect of Nannochloropsis oceanica feed inclusion on the muscle proteome of fattening rabbits. Twenty rabbits were housed in individual cages for 5 weeks and were fed a control diet (n = 10) and one with 4.45% inclusion of microalga, replacing whole soybean meal (n = 10). After slaughter, samples of gastrocnemius muscle were taken for label-free proteomic analysis. A total of 1497 proteins were identified, 46 with differential abundance. Control rabbits had high abundance of proteins related to protein metabolism, suggesting higher muscle protein turnover. They also had higher abundance of structural proteins, suggesting a less tender meat by comparison with algae-fed rabbits. These had high abundance of proteins related to amino acid catabolism (Phe, Tyr) and synthesis (Gln). In addition, they had high abundance of proteins related to protein breakdown, overall suggesting that more tender meat may result from algae feeding. BIOLOGICAL SIGNIFICANCE: Microalgae are interesting feedstuffs particularly from a sustainability point of view. Their use in farm animal diets could simultaneously enhance the nutritional value and quality of meat. However, the biochemical pathways behind such improvement remain uncharacterized, namely in the rabbit species. By using a proteomics approach, this study will enlighten such biochemical pathways and relate them with productive performance parameters. In addition, results reflect some anti-nutritional properties of the recalcitrant cell wall over amino acid digestibility and consequently, its availability for protein synthesis.

The muscular, hepatic and adipose tissues proteomes in muskox (Ovibos moschatus): Differences between males and females.

The muskox (Ovibos moschatus) is a ruminant highly adapted to arctic conditions. The objective of this work is to study liver, muscle and adipose tissues proteomes in muskoxen highlighting sex differences. Ten animals (5 per sex) were sampled in Western Greenland during the winter hunting season. During carcass processing, muscle, liver and rump fat samples were obtained. Proteomic analyses were conducted using both gel-based and gel-free approaches. Gel-free data are available (ProteomeXchange; PXD014147). For gel-free analysis, 729, 853 and 792 proteins were identified for fat, liver and muscle, respectively. Several proteins were detected with differential abundance between male and female tissues: 77, 15 and 12 proteins using gel-free for adipose tissue, liver and muscle respectively while 3 differential proteins were identified in the gel-based analysis of the adipose tissue. Females have higher abundance of proteins involved in tissue structural stability in the muscle, while males have higher abundance of proteins related to muscle development. In the liver and adipose tissue, females have higher abundance of proteins related to oxidative-stress resistance. Proteins accumulated in the adipose tissue of males highlight higher adipogenic potential. Sex dimorphism is inherent to this species, with higher abundance of proteins in specific metabolic pathways. SIGNIFICANCE: The proteomes of the muskox muscle, hepatic and adipose tissues are characterized for the first time. In addition, the effect of sex on tissue protein abundance is studied. Our results reveal that sex dimorphism goes from morphology to the molecular level in this species, affecting protein, lipid and carbohydrate metabolism. This contributes for an in-depth look into sex dimorphism using proteomics which is lacking in most mammals, apart from model species. Moreover, this information has been related to nutritional status, which is particularly important when managing the muskox population and the transformation of its habitat in relation to external factors such as climate changes that can severely affect ecosystems.

Assessing mineral status in edible tissues of domestic and game animals: a review with a special emphasis in tropical regions.

Mineral status in edible tissues has been extensively studied since the beginning of the twentieth century. Most research focus on nutrition, as the earliest reports were essentially related to nutrition, animal health and mineral deficiencies. Nutrition wise, minerals are of great importance for consumers worldwide, as meat (i.e. beef, pork, chicken) and fish are major sources of protein in human diets. Nutrition gains renewed importance in the tropical context, since tropical forages are poor in minerals. This fact contributes to mineral deficiencies and impaired production performance in extensive production systems, with greater emphasis in ruminant species. In addition to nutrition, several other factors have an important impact in mineral metabolism such as geographic location, gender and species. In this article, we aim to infer on both the role in the organism and the amount present in various edible tissues of different species, either game or production animals, presenting an overall perspective in the context of tropical animal production.

Technical quality of root canal treatment performed by undergraduate students using hand instrumentation: a meta-analysis.

The technical quality of root canal treatment (RCT) may impact on the outcome. The quality of education received during undergraduate school may be linked to the quality of treatment provided in general dental practice. In this context, the aim of this systematic review was to answer the following focused questions: (i) What is the frequency of acceptable technical quality of root fillings, assessed radiographically, performed by undergraduate students? (ii) What are the most common errors assessed radiographically and reported in these treatments? For this purpose, articles that evaluated the quality of root fillings performed by undergraduate students were selected. Data were collected based on predetermined criteria. The key features from the included studies were extracted. GRADE-tool assessed the quality of the evidence. MAStARI evaluated the methodological quality, and a meta-analysis on all studies was conducted. At the end of the screening, 24 articles were identified. Overall frequency of acceptable technical quality of root fillings was 48%. From this total, 52% related to anterior teeth, 49% to premolars and 26% to molars. The main procedural errors reported were ledge formation, furcation perforation, apical transportation and apical perforation. The heterogeneity amongst the studies was high (84-99%). Five studies had a high risk of bias, eight had a moderate risk, and 11 had low risk. The overall quality of evidence identified was very low. The conclusion was that technical quality of root fillings performed by undergraduate students is low, which may reveal that endodontic education has limited achievement at undergraduate level. A plan to improve the quality of root fillings, and by extrapolation the overall quality of root canal treatment, should be discussed by the staff responsible for endodontic education and training.

Clinical Implications caused by the agenesis of the RectusAbdominal Muscle’s (MRA) inferior venter

The rectus abdominal muscle is part of the anterior abdominal wall, having three to six bellies. In only oneof the 106 dissections already made in the “Faculdade de Ciências Médicas de Minas Gerais” AnatomyLaboratory was found a male cadaver who did not have inferior venter of this muscle bilaterally. Instead, at theleft side, was found a tendon that measured 5.5 cm laterally and 12 cm medially, and at the right side, therewas the same variation with a 15.5 cm length tendon, rising in the upper branch of the pubis and crest pubis.Despite being a rare variation, individuals who have showed it have increased potential for physiological andsurgical complications, in case they need interventions using inferior rectus abdominis muscle venter’s snips.

Influence of the polymorphisms of the alpha-major regulatory element HS-40 on in vitro gene expression

The α-MRE is the major regulatory element responsible for the expression of human α-like globin genes. It is genetically polymorphic, and six different haplotypes, named A to F, have been identified in some population groups from Europe, Africa and Asia and in native Indians from two Brazilian Indian tribes. Most of the mutations that constitute the α-MRE haplotypes are located in flanking sequences of binding sites for nuclear factors. To our knowledge, there are no experimental studies evaluating whether such variability may influence the α-MRE enhancer activity. We analyzed and compared the expression of luciferase of nine constructs containing different α-MRE elements as enhancers. Genomic DNA samples from controls with A (wild-type α-MRE) and B haplotypes were used to generate C-F haplotypes by site-directed mutagenesis. In addition, three other elements containing only the G→A polymorphism at positions +130, +199, and +209, separately, were also tested. The different α-MRE elements were amplified and cloned into a plasmid containing the luciferase reporter gene and the SV40 promoter and used to transiently transfect K562 cells. A noticeable reduction in luciferase expression was observed with all constructs compared with the A haplotype. The greatest reductions occurred with the F haplotype (+96, C→A) and the isolated polymorphism +209, both located near the SP1 protein-binding sites believed not to be active in vivo. These are the first analyses of α-MRE polymorphisms on gene expression and demonstrate that these single nucleotide polymorphisms, although outside the binding sites for nuclear factors, are able to influence in vitro gene expression.

Influence of the polymorphisms of the alpha-major regulatory element HS-40 on in vitro gene expression.

The alpha-MRE is the major regulatory element responsible for the expression of human alpha-like globin genes. It is genetically polymorphic, and six different haplotypes, named A to F, have been identified in some population groups from Europe, Africa and Asia and in native Indians from two Brazilian Indian tribes. Most of the mutations that constitute the alpha-MRE haplotypes are located in flanking sequences of binding sites for nuclear factors. To our knowledge, there are no experimental studies evaluating whether such variability may influence the alpha-MRE enhancer activity. We analyzed and compared the expression of luciferase of nine constructs containing different alpha-MRE elements as enhancers. Genomic DNA samples from controls with A (wild-type alpha-MRE) and B haplotypes were used to generate C-F haplotypes by site-directed mutagenesis. In addition, three other elements containing only the G-->A polymorphism at positions +130, +199, and +209, separately, were also tested. The different alpha-MRE elements were amplified and cloned into a plasmid containing the luciferase reporter gene and the SV40 promoter and used to transiently transfect K562 cells. A noticeable reduction in luciferase expression was observed with all constructs compared with the A haplotype. The greatest reductions occurred with the F haplotype (+96, C-->A) and the isolated polymorphism +209, both located near the SP1 protein-binding sites believed not to be active in vivo. These are the first analyses of alpha-MRE polymorphisms on gene expression and demonstrate that these single nucleotide polymorphisms, although outside the binding sites for nuclear factors, are able to influence in vitro gene expression.

Regulation of human alpha-globin gene expression and alpha-thalassemia.

Hemoglobin and globin genes are important models for studying protein and gene structure, function and regulation. We reviewed the main aspects of regulation of human alpha-globin synthesis, encoded by two adjacent genes (alpha(2) and alpha(1)) clustered on chromosome 16. Their expression is controlled mainly by a regulatory element located 40 kb upstream on the same chromosome, the alpha-major regulatory element, whose activity is restricted to a core fragment of 350 bp, within which several regulatory protein binding sites have been found. Natural deletions involving alpha-major regulatory element constitute a particular category of alpha-thalassemia determinants in which the alpha-globin genes are physically intact but functionally inactive.

Molecular markers for the characterization of Brazilian Cercospora caricis isolates.

Cercospora caricis is of interest as a potential mycoherbicide for control of purple nutsedge, Cyperus rotundus, which is considered to be the world's worst weed. The genetic variation of a collection of Brazilian Ce. caricis isolated from Cy. rotundus was analyzed by using RAPD, RFLP with a telomeric probe, [TTAGGG]18 and sequencing of the ITS1-5.8S-ITS2 regions of the ribosomal RNA gene. The Brazilian isolates were also compared with a Ce. caricis isolate from Florida, USA and with some other Cercospora species. A cluster of isolates from the Brazilian cerrado region was identified showing high genetic similarity. In contrast, isolates originating in other geographic regions of Brazil were less than 50% and 25% related to the former group according to similarity estimates produced from RAPD and telomeric RFLP analyses respectively. ITS sequence analysis did not support taxonomic division of the Brazilian strains, but did confirm the distant relatedness of these strains to the Ce. caricis isolate from Florida. The data indicate a need for an extensive molecular survey of Cercospora species associated with the Cyperaceae.

Nicotiana tabacum as an Experimental Host for the Study of Plant-Xylella fastidiosa Interactions.

Xylella fastidiosa causes citrus variegated chlorosis (CVC). Information generated from the X. fastidiosa genome project is being used to study the underlying mechanisms responsible for pathogenicity. However, the lack of an experimental host other than citrus to study plant-X. fastidiosa interaction has been an obstacle to accelerated progress in this area. We present here results of three experiments that demonstrated that tobacco could be an important experimental host for X. fastidiosa. All tobacco plants inoculated with a citrus strain of X. fastidiosa expressed unequivocal symptoms, consisting of orange leaf lesions, approximately 2 months after injection of the pathogen. CVC symptoms were observed in citrus 3 to 6 months after inoculation. The pathogen was readily detected in symptomatic tobacco plants by polymerase chain reaction (PCR) and phase contrast microscopy. In addition, X. fastidiosa was reisolated on agar plates in 4 of 10 plants. Scanning electron microscopy analysis of cross sections of stems and petioles revealed the presence of rod shaped bacteria restricted to the xylem of inoculated plants. The cell size was within the limit typical of X. fastidiosa.

Metabolism of glycogen in submandibular glands of rats. Alteration by NaF.

Submandibular glands of rats injected with NaF solution (10 mg F-/kg body weight) were analysed for glycogen content and phosphorylase activity after various time intervals. In contrast to what has been reported for the liver, sodium fluoride caused increased glycogen content. Phosphorylase (a and total) activity was not affected, suggesting a different mechanism of action of F- in the submandibular gland. In vivo experiments demonstrated stimulation of glycogenesis.