Effect of early feed restriction programs and genetic strain on humoral immune response production in broiler chickens.

Two experiments were conducted to compare the immunocompetence of Cobb high performance and rustic Label Rouge broilers and the influence of reduced growth rates subsequent to feed restriction on the IgY anti-bovine serum albumin (BSA) response. In the first experiment (EXP), 360 broilers were assigned to 36 cages from 1 to 42 days of age. A completely randomized design was applied in a 3 × 2 factorial arrangement, with 3 groups (Label Rouge, Cobb ad libitum, and Cobb Restricted Intake), and 2 levels of energy (3,100 and 2,800 kcal/kg); there were 6 replicates per treatment. In the second EXP, 384 Cobb 500 male broilers were randomly assigned to the following feed restriction programs from day 8 to 16: Control, fed ad libitum; Quantitative (80% of the control amount); By Time (fed for 8 h/d), and Qualitative (80% limiting nutrients) restriction. Blood samples were collected on days 35 and 42 (EXP 1) and weekly from day 7 to 42 (EXP 2) for IgY anti-BSA quantification. In EXP 1, the production of IgY anti-BSA was lower in the Cobb groups (P < 0.0001) than in the Label Rouge group, and higher in the Cobb Restricted Intake group (P < 0.0001) compared with the same genetic strain fed ad libitum. Birds fed the low energy diet presented lower (P ≤ 0.06) IgY anti-BSA, independent of genetics. In EXP 2, no difference (P > 0.05) was observed 1 wk after the first BSA inoculation. However, at day 28, birds in all feed restriction programs had higher (P < 0.05) IgY anti-BSA than the Control group fed ad libitum. At day 35, the greatest residual effect of IgY anti-BSA was observed in the Quantitative restriction group. No differences (P > 0.05) were observed between groups after 42 d. The 3 early feed restriction programs had beneficial effects on the humoral immune response. Overall, Quantitative restriction promoted a longer lasting IgY anti-BSA response. Lower growth rate, due to feed restriction or genetic potential, improves humoral immunity in broiler chickens.

Infant feeding practices and determinant variables for early complementary feeding in the first 8 months of life: results from the Brazilian MAL-ED cohort site.

OBJECTIVE: The present study aimed to describe breast-feeding, complementary feeding and determining factors for early complementary feeding from birth to 8 months of age in a typical Brazilian low-income urban community. DESIGN: A birth cohort was conducted (n 233), with data collection twice weekly, allowing close observation of breast-feeding, complementary feeding introduction and description of the WHO core indicators on infant and young child feeding. Infant feeding practices were related to socio-economic status (SES), assessed by Water/sanitation, wealth measured by a set of eight Assets, Maternal education and monthly household Income (WAMI index). Two logistic regression models were constructed to evaluate risk factors associated with early complementary feeding. RESULTS: Based on twice weekly follow-up, 65 % of the children received exclusive breast-feeding in the first month of life and 5 % in the sixth month. Complementary feeding was offered in the first month: 29 % of the children received water, 15 % infant formulas, 13 % other milks and 9·4 % grain-derived foods. At 6 months, dietary diversity and minimum acceptable diet were both 47 % and these increased to 69 % at 8 months. No breast-feeding within the first hour of birth was a risk factor for the early introduction of water (adjusted OR=4·68; 95 % CI 1·33, 16·47) and low WAMI index a risk factor for the early introduction of other milks (adjusted OR=0·00; 95 % CI 0·00, 0·02). CONCLUSIONS: Data suggest local policies should promote: (i) early breast-feeding initiation; (ii) SES, considering maternal education, income and household conditions; (iii) timely introduction of complementary feeding; and (iv) dietary diversity.

Analysis of Mendelian inheritance and genetic linkage in microsatellite loci of Eucalyptus urophylla S.T. Blake.

Eucalyptus urophylla is an important species in the Brazilian forest sector due to its rapid growth rates and resistance to disease. The aim of this study was to verify Mendelian inheritance, genetic linkage, and genotypic disequilibrium for 15 microsatellite loci, with the goal of producing a robust set of genetic markers. Mendelian inheritance and genetic linkage analyses were carried out using genotypes from maternal trees, and their open-pollinated seeds and genotypic disequilibrium were assessed using adult trees. By comparing heterozygous maternal genotypes and their seeds, we found no significant deviations from the expected 1:1 Mendelian segregation and the expected 1:1:1:1 segregation hypothesis for pairwise loci. For adult trees, we did not find strong evidence of genotypic imbalance for pairwise loci. Our results indicated that the analyzed set of microsatellite loci could be used to carry out analyses of genetic diversity, mating system, and parentage in E. urophylla.

Mendelian inheritance, genetic linkage, and genotypic disequilibrium for nine microsatellite loci in Cariniana estrellensis (Raddi) Kuntze (Lecythidaceae).

Cariniana estrellensis is one of the largest trees found in Brazilian tropical forests. The species is typical of advanced stages of succession, characteristic of climax forests, and essential in genetic conservation and environmental restoration plans. In this study, we assessed Mendelian inheritance, genetic linkage, and genotypic disequilibrium in nine microsatellite loci for a C. estrellensis population. We sampled and genotyped 285 adult trees and collected seeds from 20 trees in a fragmented forest landscape in Brazil. Based on maternal genotypes and their seeds, we found no deviation from the expected 1:1 Mendelian segregation and no genetic linkage between pairwise loci. However, for adults, genotypic disequilibrium was detected for four pairs of loci, suggesting that this result was not caused by genetic linkage. Based on these results, we analyzed microsatellite loci that are suitable for use in population genetic studies assessing genetic diversity, mating system, and gene flow in C. estrellensis populations.

Short Communication Mendelian inheritance, linkage, and genotypic disequilibrium in microsatellite loci of Hymenaea stigonocarpa Mart. ex Hayne (Fabaceae-Caesalpinioideae).

Hymenaea stigonocarpa is a deciduous and monoecious Neotropical tree species pollinated by bats. Due to overexploitation and habitat destruction, the population size has drastically diminished in nature. No previous study has investigated Mendelian inheritance, linkage, and genotypic disequilibrium in the available microsatellite markers in this species. So, our aim was to estimate these parameters using six microsatellite loci in a sample of 470 adults and 219 juveniles from two populations of H. stigonocarpa. In addition, 30 seeds per tree from 35 seed-trees were collected. Each seed was kept record of the seed-trees and fruit origin. Based on the six microsatellite loci, we found that only 10.6% of the cases showed significant deviations from Mendelian segregation and 15.3% showed linkage. We detected no evidence of genotypic disequilibrium between the loci in the adult trees or juveniles. Thus, our results suggest that these loci can be used with great accuracy in future genetic analyses of H. stigonocarpa populations.

Immunosensor for diagnosis of Alzheimer disease using amyloid-ß 1-40 peptide and silk fibroin thin films.

Layer-by-Layer (LbL) films containing silk fibroin (SF) and the 40 aminoacid-long amyloid-ß peptide (Aß1-40) were prepared with the purpose of developing a new prototype of an electrochemical immunosensor. The film showed a satisfactory growth in quartz substrate and screen-printed carbon electrodes, as observed by UV-vis spectroscopy and cyclic voltammetric, respectively. The peptide immobilized in LbL films in junction with SF shows secondary structure induced, as shown by circular dichroism measurements, favoring the interaction SF/peptide LbL film with the specific antibody. Immunosensor showed a linear response in the presence of the antibody with concentrations from 0 to 10ngmL(-1) both analyzed by current changes in 0.3V and voltammogram area. This system can be applied as a new prototype for preliminary diagnosis of Alzheimer's disease.

Effects of conjugated linoleic acid and lutein on the growth performance and immune response of broiler chickens.

The effects of lutein and conjugated linoleic acid (CLA) on growth performance and immune response of broiler chickens were evaluated in the presence and absence of Salmonella lipopolysaccharide (LPS) immune challenge. Cobb chicks (360; 1 to 22 d of age) were used in a 3 × 2 factorial arrangement of CLA (0, 1, and 2%) and lutein (0 and 50 mg/kg) dietary levels. At d 8 and 15, birds were injected with BSA to assess IgY production. At d 20, birds were injected with LPS. Samples of liver, spleen, and duodenum were collected at 3 and 16 h post-LPS challenge for RT-qPCR analysis of RXRα, RXRγ, PPARα, PPARγ, TLR-4, IL-1ß, IL-2, IL-10, and IL-12 gene expression. CLA decreased BW, BW gain (BWG), and G:F from d 1 to 20, but these effects were reversed when lutein was included in the 1% CLA diet (P < 0.001). The production of IgY anti-BSA increased following a 2% CLA supplementation (P < 0.01). LPS increased the liver:BW ratio at 3 h post-injection (P < 0.001) and decreased BWG at 3, 16, and 40 h (P < 0.001). Lutein decreased plasmatic nitric oxide levels (P < 0.01). LPS downregulated PPARα mRNA in the duodenum (P = 0.02) and liver (P = 0.04), and PPARγ (P = 0.01) and RXRα (P = 0.08) in the spleen; these effects were not reversed by CLA or lutein as initially hypothesized. Although LPS upregulated IL-1ß (P = 0.02) and IL-12 (P = 0.07) expression, lutein downregulated these pro-inflammatory cytokines in the liver (P = 0.03 and P = 0.07, respectively). Lutein decreased splenic (P = 0.09) but increased hepatic (P = 0.06) TLR-4 mRNA. A dietary CLA supplementation of 2% increased hepatic RXRα (P = 0.10). In conclusion, CLA decreased broiler chicken growth performance, but lutein could prevent this negative effect (depending on CLA dose). Lutein had an anti-inflammatory effect, and a 2% CLA supplementation improved the humoral immune response.

Mendelian inheritance, genetic linkage, and genotypic disequilibrium at microsatellite loci in Genipa americana L. (Rubiaceae).

Genipa americana is a tropical tree species that is widely distributed in the humid tropical and subtropical regions of Central and South America. This study investigated Mendelian inheritance, genetic linkage, and genotypic disequilibrium at six microsatellite loci developed for G. americana. Adult trees (188) and regenerants (163) were sampled and genotyped in a fragmented population of the species. We also genotyped open-pollinated seeds from 12 seed-trees during reproductive events in 2010 and 2011. Significant deviations from the expected 1:1 Mendelian segregation were detected in 29.5% of the tests. Significant genetic linkage between pairwise loci was detected in 54.4% of the tests, but no genotypic disequilibrium was detected between pairwise loci for adult trees and regenerants. Overall, the results indicate that the six loci analyzed may be used in studies of G. americana's genetic diversity and structure, its mating system, and in parentage analyses.

Amperometric glucose biosensor based on layer-by-layer films of microperoxidase-11 and liposome-encapsulated glucose oxidase.

An important step in several bioanalytical applications is the immobilization of biomolecules. Accordingly, this procedure must be carefully chosen to preserve their biological structure and fully explore their properties. For this purpose, we combined the versatility of the layer-by-layer (LbL) method for the immobilization of biomolecules with the protective behavior of liposome-encapsulated systems to fabricate a novel amperometric glucose biosensor. To obtain the biosensing unit, an LbL film of the H2O2 catalyst polypeptide microperoxidase-11 (MP-11) was assembled onto an indium-tin oxide (ITO) electrode followed by the deposition of a liposome-encapsulated glucose oxidase (GOx) layer. The biosensor response toward glucose detection showed a sensitivity of 0.91±0.09 (µA/cm2)/mM and a limit of detection (LOD) of 8.6±1.1 µM, demonstrating an improved performance compared to similar biosensors with a single phospholipid-liposome or even containing a non-encapsulated GOx layer. Finally, glucose detection was also performed in a zero-lactose milk sample to demonstrate the potential of the biosensor for food analysis.

Effect of CLA on performance and immune response of weanling piglets.

An experiment was conducted to evaluate the effect of CLA on the immune response and performance of piglets when subjected to an immune challenge. A total of 32 weanling pigs (17 to 23 d of age) with an initial BW of 8.9 kg were allotted to a 3 × 2 factorial arrangement of treatments. There were 3 levels of dietary CLA (0%, 1%, and 2%) and 2 levels of lipopolysaccharide (LPS) challenge (unchallenged and challenged). Challenged pigs were challenged on d 7 and 21. On d 4 and 18, all pigs were inoculated with BSA for assessment of IgG production. There was no difference in growth performance among piglets receiving different CLA supplementation levels. However, LPS-challenged piglets had poorer BW (P < 0.05), ADFI (P < 0.01), and ADG (P < 0.001) compared with the control group at d 35 postweaning. Lipopolysaccharide-challenged piglets also had increased respiratory rate (P < 0.001) and rectal temperature (P < 0.001), and decreased plasma proteins, hematocrit, and white blood cell counts (P < 0.05). Production of IgG against BSA was increased in the 1% CLA supplementation group (P < 0.001), indicating that CLA has an immunomodulatory effect. Supplementation with CLA did not affect lymphocyte proliferation, percentage of CD4(+) and CD8(+) cells, plasma proteins, red and white blood cell count, respiratory rate, or rectal temperature after LPS challenge. Although CLA supplementation did not influence growth performance or certain immune system measurements, the increased IgG titers with 1% CLA dietary supplementation indicate that it has a beneficial effect on the humoral immune system of weaned piglets.

Low levels of realized seed and pollen gene flow and strong spatial genetic structure in a small, isolated and fragmented population of the tropical tree Copaifera langsdorffii Desf.

Over the past century, the Brazilian Atlantic forest has been reduced to small, isolated fragments of forest. Reproductive isolation theories predict a loss of genetic diversity and increases in inbreeding and spatial genetic structure (SGS) in such populations. We analysed eight microsatellite loci to investigate the pollen and seed dispersal patterns, genetic diversity, inbreeding and SGS of the tropical tree Copaifera langsdorffii in a small (4.8 ha), isolated population. All 112 adult trees and 128 seedlings found in the stand were sampled, mapped and genotyped. Seedlings had significantly lower levels of genetic diversity (A=16.5±0.45, mean±95% s.e.; H(e)=0.838±0.006) than did adult trees (A=23.2±0.81; H(e)=0.893±0.030). Parentage analysis did not indicate any seed immigration (m(seeds)=0) and the pollen immigration rate was very low (m(pollen)=0.047). The average distance of realized pollen dispersal within the stand was 94 m, with 81% of the pollen travelling <150 m. A significant negative correlation was found between the frequency and distance of pollen dispersal (r=-0.79, P<0.01), indicating that short-distance pollinations were more frequent. A significant SGS for both adults (∼50 m) and seedlings (∼20 m) was also found, indicating that most of the seeds were dispersed over short distances. The results suggested that the spatial isolation of populations by habitat fragmentation can restrict seed and pollen gene flow, increase SGS and affect the genetic diversity of future generations.

Determination of polyamines in organic extracts from roots of Canavalia ensiformis by capillary electrophoresis.

A rapid, selective and specific capillary zone electrophoresis method to determine polyamines in organic extracts from roots of Canavalia ensiformis (Jack Beans) was developed using ultra violet (UV) detection. Canavalia ensiformis is relatively free from diseases and it is used as reference in allelopathy studies. Polyamines are widely distributed in plant and it could be involved in plant pathogen interactions. Optimal separation was achieved using 15 mmol L(-1)formic acid (pH 3.0) + 4 mmol L(-1) imidazole as a background electrolyte. It was possible to identify and quantify the polyamines on herbal samples in the presence of other phytochemical substances and analyze them quickly (up to 6 min). The applicability of this method was evaluated in crude organic extracts from roots of Canavalia ensiformis.

Regulatory CD4 T cells: expression of IL-2R alpha chain, resistance to clonal deletion and IL-2 dependency.

We recently characterized a CD4+ T cell population expressing the IL-2R alpha chain (CD25), producing IL-10 and resisting clonal deletion induced by viral superantigen (vSAG) encoded by mouse mammary tumor virus [MMTV(SW)]. We now report that these apoptosis-resistant cells are generated in the thymus but not from the immature CD4+ CD8+ thymocytes. They migrate from the thymus and are found in the periphery from at least the 10th day of life, after which they expand with the same kinetics in normal and MMTV(SW)-infected mice. Their strong capacity for expansion in the periphery makes this population insensitive to thymectomy in adulthood. CD4+ CD25+ cells were totally dependent on exogenous IL-2 for growth in vitro and in vivo, and were missing in IL-2 knockout (KO) mice. The absence of this population and/or an inability to produce IL-10 may be the missing link between IL-2R alpha KO, IL-2 KO and IL-10 KO mice, which all die of inflammatory bowel disease.