Insights on bovine genetic engineering and cloning

Transgenic technology has become an essential tool for the development of animal biotechnologies, and animal cloning through somatic cell nuclear transfer (SCNT) enabled the generation of genetically modified animals utilizing previously modified and selected cell lineages as nuclei donors, assuring therefore the generation of homogeneous herds expressing the desired modification. The present study aimed to discuss the use of SCNT as an important methodology for the production of transgenic herds, and also some recent insights on genetic modification of nuclei donors and possible effects of gene induction of pluripotency on SCNT.

Life history, biomass and production of Coronatella rectangula (Branchiopoda, Anomopoda, Chydoridae) from Minas Gerais

Species of Chydoridae provide the main diversity of the Cladocera. These organisms have been the subject of many studies; some dealing with their role in energy flow in aquatic ecosystems, since they inhabit the littoral region of water bodies which undergo the first impacts from anthropic activities. The aim of this study is to increase knowledge about the life cycle of Coronatella rectangula (Sars, 1861), a species found in several water bodies in the state of Minas Gerais, Brazil. The life cycle was determined by the culture of parthenogenetic females under controlled conditions in the laboratory. Experimental cultures were maintained in growth chambers at a constant temperature of 23.6(±0.5)ºC, through a 12 h light/12 h dark photoperiod. The organisms were fed on a suspension of Pseudokirchneriella subcapitata (Chlorophyceae) (10(5) cells.mL-1), and 0.02 mL of a mixed suspension of yeast and fish ration added per organism in equal proportions (1:1). Fifty parthenogenetic females with eggs were isolated and maintained until they produced neonates. Thirty of these neonates that had less than 24 hours were put in polypropylene bottles of 50 mL and kept in a germination chamber. These organisms were observed daily to obtain the parameters of the life cycle. Biomass and secondary production were also calculated. The embryonic development time of the specimens of C. rectangula was 1.68(±0.13) days and the time to reach primipara, was 2.48(±0.45) days. The mean fecundity of C. rectangula was two eggs/female/brood and the total number of eggs produced by the female during its life cycle was 27.8 eggs. During the whole life cycle, specimens of C. rectangula had a maximum of 14 seedlings, with two instars in the juvenile stage. Total biomass for C. rectangula was 36.66 µgDW.m-3(9.83 for the juvenile stage and 26.82 µgDW.m-3 for adults), and secondary production was 12.10 µgDW.m-3.day-1(8.34 µgDW.m-3.day-1 for egg production and 3.76 µgDW.m-3.day-1 for the juvenile stage).

The use of monoclonal antibodies for the characterization and production of Mycobacterium leprae antigens

Similar immunizations of mice and hybridoma technology were used by several investigators to raise monoclonal antibodies which identified a limited range of epitopes and antigenic molecules. Further studies would have the scope for revealing yet more novel structures. The existing MABs are agreed standard reagents, avaiable to investigators and valuable for several applications. At least six epitopes specific for M. leprae were defined in molecular terms. Monoclonal antibody based immunoassays proved to be invaluable for the screening of recombinant DNA clones and for the topographic study of individual epitopes. Purification of antigens using affinity chromatography requires further development of techniques whilst serology of leprosy is open for clinical and epidemiological evaluation.

Penicillin tolerance among Beta-hemolytic streptococci and production of the group carbohydrates, hemolysins, hyaluronidases and deoxyribonucleases

Penicillin tolerance among 67 strains of beta-hemolytic streptococci was examined by determining the ratio of the minimal bactericidal concentration to the minimal inhibitory concentration as 32 or greater. Tolerance was demonstrated in 15 group A strains and in 11,7, and 4 of groups B, C and G, respectively. Thereafter the effects of a subminimal inhibitory concentration (1/2MIC) of penicillin on the bacterial products of four tolerant and four nontolerant strains (two of each Lancefield group) were analyzed and compared. The antibiotic caused a marked increase in the expression of the group carbo-hydrates for strains of group B. Penicillin was found to reduce the cell-bound hemolysin activities of the four tolerant strains and to increase the activity of the other (free) form of nontolerant groups A, C and G hemolysins. Penicillin caused an increase in the extracellular hyaluronidase activities of one group A and groups B, C and G streptococci. With added antibiotic the production of deoxyribonuclease by tolerant groups A, C and G was greatly enhanced and that of the group B streptococcus was arrested.

Management of slips and its effect on growth and production of 'Pérola' pineapple plants

The lack of good quality planting material has limited the expansion and contributed to yield reduction of the Brazilian pineapple culture. Alternatives of 'Pérola' pineapple slips management were studied aiming at obtaining superior planting material within a shorter time period and making good use of healthy slips of low vigor, that are commonly discarded by growers. Two experiments were carried out at the Experimental Field of Embrapa Cassava & Fruits, Cruz das Almas, Bahia, Brazil, and another one in a commercial plantation in the region of Itaberaba, BA, using blocks or entirely randomized designs with at least four replications. In the first one, the development of slips of different initial sizes (6 to 20 cm long), when grown on mother plants after fruit harvest, was compared with that of slips grown in a nursery after their removal from the mother plants. It became clear that larger slips grow more vigorously and that the removal from the mother plant delays their growth. However, results from the second study showed that those slips grown in nursery had vegetative and agronomic performance similar to that of conventional slips and close to that of plantlets produced from plant stem sections. Independently from the type of planting material used, the larger ones presented a more vigorous growth and produced higher yields. In the third study, it was observed the influence of mineral fertilization, pest control and growth regulator application after fruit harvest on slip development. The treatments applied did not significantly accelerate slip growth. Slips reached 50 cm length and at least 300 g fresh weight within 90 days after fruit harvest, indicating that vigorous plants have enough nutritional storage material for slips development.

Growth and production of irrigated vitória pineapple grown in semi-arid conditions

This study aimed to evaluate the growth characteristics of irrigated Vitória pineapple plants grown in semi-arid conditions and determine its developmental stages based on those characteristics. It was used a randomized block design with four replicates. The experimental treatments were: plant harvest at 270, 330, 390, 450, 510, 570, 690, 750, and 810 days after planting (DAP). The following variables were determined: plant height, stem diameter, D-leaf length, D-leaf fresh and dry mass, biomass production of plants and plant parts (organs), and vegetative biomass. Five phenological stages are proposed based on vegetative biomass production: < 20% biomass production (V1); 21-40% (V2); 41-60% (V3); 61-80% (V4); and > 80% (V5). The maximum growth rate for plant height, D-leaf length, and stem diameter was observed at the end of the phenological stage V1 (390-411 DAP), and at the end of stage V5 these plant traits had average values of 106, 82, and 7 cm, respectively. The maximum biomass accumulation rates were observed at stages V4 and V5, resulting in a final fruit yield and total fresh biomass of 72 t ha-1 and 326 t ha-1, respectively. Finally, we estimated that 80% of the accumulated biomass may remain in the field after fruit and slip harvest, and could be incorporated as plant residue into the soil.

Expression of Grapevine leafroll-associated virus 3 coat protein gene in Escherichia coli and production of polyclonal antibodies

Grapevine leafroll-associated virus 3 (GLRaV-3), the main viral species of the grapevine leafroll complex, causes yield and quality reduction in grapes (Vitis spp.). The coat protein gene was RT-PCR-amplified from total RNA extracted from infected grapevine leaves and the amplified fragment was cloned and completely sequenced. The fragment was subsequently subcloned into the pRSET-C expression vector. The recombinant plasmid was used to transform Escherichia coli BL21:DE3 and express the capsid protein. The coat protein, fused to a 6 His-tag, was purified by affinity chromatography using an Ni-NTA resin. The identity of the purified protein was confirmed by SDS-PAGE and Western blot. The in vitro-expressed protein was quantified and used for rabbit immunizations. The antiserum was shown to be sensitive and specific for the detection of GLRaV-3 in grapevine extracts in Western blot and DAS-ELISA assays, with no unspecific or heterologous reactions against other non-serologically related viruses being observed.

Effect of light intensity and growth substratum on plant development and production of secondary metabolites in Cordia curassavica (Jacq.) Roem. & Schult

Cordia curassavica (Jacq.) Roem. & Schult. (Boraginaceae), also referred to as Cordia verbenacea DC, has been traditionally used for medicinal purposes. This study was driven to verify the behavior of the species in similar conditions to its natural environment, such as high light intensity and sandbank soil, and in conditions of low light intensity and fertilized substratum (dystroferric red nitosoil plus earthworm humus). The growth of the plant, the income of leaf crude extracts and, in the alcoholic extract, the number of substances found in thin layer cromatography and the toxicity of the substratum was observed. The results indicated that the growth of the root biomass, stem and leaves in discharge or lower light intensity was similar, but smaller in sandbank soil than in fertilized soil. The relative income of extracts in ether of petroleum and alcohol was larger in high light intensity and fertilized substratum. The light intensity and the substratum type didn't affect the number of substances detected in the alcoholic extract or the toxicity of this extract. Stains corresponding to the rosmarinic acid were only evidenced in some samples of the alcoholic extract, not allowing the verification of the effect of the treatments about its production.

POPULATION STRUCTURE AND PRODUCTION OF COPAIBA OLEORESIN BETWEEN VALLEYS AND HILLSIDES OF THE MINING AREA OFTROMBETAS RIVER - PARÁ1

ABSTRACT We aimed in this work to study natural populations of copaiba (Copaifera multijuga Hayne) on the Monte Branco mountain at Porto Trombetas-PA, in order to support sustainable management and the exploitation of oleoresin from copaiba. We studied the population structure of copaiba on hillsides and valleys of the south face of Monte Branco, within Saracá Taquera National Forest, where bauxite ore was extracted in the biennium 2013-2014 by Mineração Rio do Norte (MRN). We produced a 100% forest inventory of the specie and of oleoresin extraction in order to quantify the potential production of the remaining area. The density of copaiba individuals with DBH > 30 cm was 0.33 individuals per hectare in the hillside and 0.25 individuals per hectare in the valley. Both environments presented a density of 0.28 individuals per hectare. The average copaiba oleoresin yield was 0.661±0.334 liters in the hillside and 0.765±0.280 liters in the valley. The average value of both environments together (hillside and valley) was 0.714±0.218 liters. From all individuals with DBH over 30 cm, 38 (58%) produced some amount of oleoresin, averaging 1.113±0.562 liters in the hillside, 1.329±0.448 liters in the valley and 1.190±0.355 liters in both environments together. The results show the need for planning the use of the surroundings of the study area in order to reach the required volume of copaiba to make feasible the sustainable management of oleoresin extraction in the region.

Adhesion and production of degrading enzymes by bacteria isolated from biofilms in raw milk cooling tanks

Biofilms in milk cooling tanks compromise product quality even on farms. Due to the lack of studies of this topic, this study evaluated the microbiological conditions of raw milk cooling tanks on farms and characterized the microorganisms isolated from these tanks. Samples were wiped off with sterile swabs from seven milk cooling tanks in three different points in each tank. Mesophiles and psychrotrophic counts were performed in all samples. The isolation of Pseudomonas spp., Bacillus cereus and atypical colonies formed on selective media were also performed, totalizing 297 isolates. All isolates were tested for protease and lipase production and biofilm formation. Of the total isolates, 62.9% produced protease, 55.9% produced lipase, and 50.2% produced biofilm. The most widespread genus inside the milk cooling tank was Pseudomonas since it was not possible to associate this contamination with a single sampling point in the equipment. High counts of microorganisms were found in some cooling tanks, indicating poor cleaning of the equipment and providing strong evidences of microbial biofilm presence. Moreover, it is worth mentioning the milk potential contamination with both microbial cells and their degrading enzymes, which compromises milk quality.

Evaluation of roof slope and exposure with different roofing materials in reduced models of animal production facilities in spring and summer

The present study was conducted at the Department of Rural Engineering and the Department of Animal Morphology and Physiology of FCAV/Unesp, Jaboticabal, SP, Brazil. The objective was to verify the influence of roof slope, exposure and roofing material on the internal temperature of reduced models of animal production facilities. For the development of the research, 48 reduced and dissemble models with dimensions 1.00 × 1.00 × 0.50 m were used. The roof was shed-type, and the models faced to the North or South directions, with 24 models for each side of exposure. Ceramic, galvanized-steel and fibro tiles were used to build the roofs. Slopes varied between 20, 30, 40 and 50% for the ceramic tile and 10, 30, 40 and 50% for the other two. Inside the models, temperature readings were performed at every hour, for 12 months. The results were evaluated in a general linear model in a nested 3 × 4 × 2 factorial arrangement, in which the effects of roofing material and exposure were nested on the factor Slope. Means were compared by the Tukey test at 5% of probability. After analyzing the data, we observed that with the increase in the slope and exposure to the South, there was a drop in the internal temperature within the model at the geographic coordinates of Jaboticabal city (SP/Brazil).

Accountability and co-production of information and control: social observatories and their relationship with government agencies

Abstract:Through the development of a proposal to categorize accountability into four stages - classical, cross-sectional, systemic, and diffused -, this article aims to identify characteristics of co-production of information and socio-political control of public administration in the work of Brazilian social observatories in relationship with government control agencies. The study analyses data from 20 social observatories and, particularly, three experiences of co-production of information and control, based on a systemic perspective on accountability and a model with four categories: Political and cultural; valuing; systemic-organizational, and production. The conclusions summarize characteristics of these practices, specific phases in the accountability processes, as well as the potentialities and challenges of co-production of information and control, which not only influences, but it is also influenced by the accountability system.

Analysis of the expression of toll-like receptors 2 and 4 and cytokine production during experimental Leishmania chagasi infection

Toll-like receptors (TLRs) recognise pathogen-derived molecules and influence immunity to control parasite infections. This study aimed to evaluate the mRNA expression of TLRs 2 and 4, the expression and production of the cytokines interleukin (IL)-12, interferon (IFN)-γ, tumor necrosis factor (TNF)-α, IL-17, IL-10 and transforming growth factor (TGF)-β and the production of nitric oxide (NO) in the spleen of mice infected with Leishmania chagasi. It also aimed to evaluate any correlations between mRNA expression TLR2 and 4 and cytokines and NO production. Infection resulted in increased TLR2-4, IL-17, TNF-α and TGF-β mRNA expression during early infection, with decreased expression during late infection correlating with parasite load. IFN-γ and IL-12 mRNA expression decreased at the peak of parasitism. IL-10 mRNA expression increased throughout the entire time period analysed. Although TGF-β, TNF-α and IL-17 were highly produced during the initial phase of infection, IFN-γ and IL-12 exhibited high production during the final phase of infection. IL-10 and NO showed increased production throughout the evaluated time period. In the acute phase of infection, there was a positive correlation between TLR2-4, TNF-α, IL-17, NO, IL-10 and TGF-β expression and parasite load. During the chronic phase of infection, there was a positive correlation between TLR2-4, TNF-α, IL-17 and TGF-β expression and parasite load. Our data suggest that infection by L. chagasi resulted in modulation of TLRs 2 and 4 and cytokines.

Coffea arabica (cv Catuaí) production and bean size under different population arrangements and soil water availability

The purpose of this research was to evaluate the effect of drip irrigation under different population arrangements on the phytometric features, coffee productivity and bean size classification according to sieve retention. The experiment with Coffea arabica L. cv Catuaí was carried out in Mococa, São Paulo, Brazil. The experimental design was a 6 x 2 factorial scheme in randomized blocks, with four replications. The six densities of plantation were E1 (1.60 x 0.50 m); E2 (1.60 x 0.75 m); E3 (1.60 x 1.00 m); E4 (3.20 x 0.50 m); E5 (3.20 x 0.75 m) and E6 (3.20 x 1.00 m), which were divided according to the availability of water (irrigated - I - or non-irrigated - NI - groups). Data were submitted to analysis of variance and averages compared by Tukey test at 1 and 5% of probability. Descriptive analysis of coffee beans according to sieve classification was performed. Irrigation promoted an increase in plant height, crown diameter and production of processed coffee when compared with the NI group. Interaction between population arrangement and irrigation was observed, with an increase in production and crown diameter as the spacing was decreased. Therefore, irrigation provided significant increase in coffee bean size.