Molecular tools and analytical approaches for the characterization of farm animal genetic diversity

Genetic studies of livestock populations focus on questions of domestication, within- and among-breed diversity, breed history and adaptive variation. In this review, we describe the use of different molecular markers and methods for data analysis used to address these questions. There is a clear trend towards the use of single nucleotide polymorphisms and whole-genome sequence information, the application of Bayesian or Approximate Bayesian analysis and the use of adaptive next to neutral diversity to support decisions on conservation.

Gene expression in placentation of farm animals: An overview of gene function during development

Eutherian mammals share a common ancestor that evolved into two main placental types, i.e., hemotrophic (e.g., human and mouse) and histiotrophic (e.g., farm animals), which differ in invasiveness. Pregnancies initiated with assisted reproductive techniques (ART) in farm animals are at increased risk of failure; these losses were associated with placental defects, perhaps due to altered gene expression. Developmentally regulated genes in the placenta seem highly phylogenetically conserved, whereas those expressed later in pregnancy are more species-specific. To elucidate differences between hemotrophic and epitheliochorial placentae, gene expression data were compiled from microarray studies of bovine placental tissues at various stages of pregnancy. Moreover, an in silico subtractive library was constructed based on homology of bovine genes to the database of zebrafish - a nonplacental vertebrate. In addition, the list of placental preferentially expressed genes for the human and mouse were collected using bioinformatics tools (Tissue-specific Gene Expression and Regulation [TiGER] - for humans, and tissue-specific genes database (TiSGeD) - for mice and humans). Humans, mice, and cattle shared 93 genes expressed in their placentae. Most of these were related to immune function (based on analysis of gene ontology). Cattle and women shared expression of 23 genes, mostly related to hormonal activity, whereas mice and women shared 16 genes (primarily sexual differentiation and glycoprotein biology). Because the number of genes expressed by the placentae of both cattle and mice were similar (based on cluster analysis), we concluded that both cattle and mice were suitable models to study the biology of the human placenta. (C) 2011 Elsevier B.V. All rights reserved.

Effects of a small fish cage farm on zooplankton assemblages (Cladocera and Copepoda: Crustacea) in a sub-tropicalreservoir (SE Brazil).

In Brazil, the environmental impacts of fish cage farming in water reservoirs have not been well studied. As this activity is being increasingly practiced, investigations on the impacts of this practice are strongly needed. The goal of this study was to investigate the effects of a small cage tilapia farm on zooplankton assemblages in an oligo/mesotrophic reservoir (Jurumirim Reservoir, Paranapanema River). Zooplankton, limnological variables, and water samples were obtained trimonthly during a year at two sample sites, one was located adjacent to the cage farm and the control area was located one kilometer away from it. Eighteen species were identified and Cladocera was the dominant group. The same species of microcrustaceans were identified at both sites. Among the ecological attributes studied, only evenness showed a tendency towards being higher in the control site. Significant differences between studied variables in the sites were observed only for material in suspension. The results of the study indicate that, during the studied period, the cage farm did not generate detectable changes in the zooplankton assemblages and their ecological attributes. However, small differences in some limnological variables could be an indication of some environmental changes associated with the fish farm system.

Pollen viability in Syagrus romanzoffiana and S. coronata (Arecaceae)

Palms of the Syagrus genus have several natural hybrids in Brazil. They are widely cultivated around the world as ornamental palm. S. romanzqffiana is the parental of natural hybrids with other Syagrus species that are potentially useful as palm cultivated for heart of palm production. The objective of this work was to evaluate the pollen grains viability got from plants grown in the Experimental Nursery of the FCAVJ/UNESP, Jaboticabal, SP, Brazil. Pollen grains were sampled from inflorescence in three occasions: in the right day of anthesis, one day before and one day after anthesis. For each day, the inflorescence was subdivided in apical, medium and basal portions where pollen grains were collected. The results shown that: a) for S. coronata there was no statistical differences in pollen viability (pollen viability average = 92.89%); b) for S. romanzoffiana the average pollen viability was 87.27%, 87.47% e 86.85% before, during and after the anthesis, respectively. The basal portion pollen grains had lower viability (85.19% - p<0.01) than those from the medium (88.17%) and the apical (88.46%) portion of the inflorescence. These results shown that both species have no problems with pollen viability and can be used as parental of hybrids with other Syagrus species.

Economic viability and selection of irrigation systems using simulation and stochastic dominance

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Avaliação do perfil eletroforético das proteínas séricas em matrizes pesadas (gallus gallus domesticus) da linhagem avian farm

This work is aimed to determine the profile of electrophoretic serum protein in healthy adult broiler breeders (Gallus gallus domesticus) of the Avian farm strain. Fifteen breeders aging 63 weeks from Conchas, city located in the State of São Paulo, were assessed. The biuret method was used to obtain the total serum protein values and protein fractions separation through electrophoresis technique in agarose gel, and film reading through densitometry in 520nM. Seven fractions were obtained, whereas, β 1 - globulin and β 2 - globulin were not cited by the authors in the textbooks checked. The prealbumin fraction was identified only in six out of 15 samples analyzed. In five breeders, it was observed the division of g - globulin into two fractions named g - 1 and g - 2, according to the electrophoretic mobilities. The relation albumin/globulin (A/G) found in the experiment agrees with the other authors cited, demonstrating that it decreases as the age increases.

Viability of Streptococcus mutans on transparent and opaque toothbrushes.

PURPOSE: The objective of this study was to evaluate the viability of the microorganism Streptococcus mutans on toothbrushes made of opaque and transparent materials. METHODS: Twenty-eight toothbrushes (14 opaque and 14 transparent) were inoculated in tubes with brain heart infusion (BHI) broth of a standard strain of S. mutans and incubated in candle jars at 37 degrees C for 24 hours. Both the opaque and transparent toothbrushes were removed at T = 0 h (control); T = 0.5 h; T = 1 h; T = 2 h; T = 4 h; T = 8 h; and T = 24 h. Individual toothbrushes were subjected to agitation in a saline solution and samples of the solution were diluted and inoculated in Bacitracin Sucrose Agar--SB-20. RESULTS: After half an hour (T2) there was a significant decrease in the number of microorganisms on the transparent and opaque toothbrushes, respectively 6.0 x 10(5) and 9.4 x 10(5), when compared to the control. After the T3 = 1 hour, T4 = 2 hours, T5 = 4 h, the number of microorganisms decreased from 4.1 x 10(5); 2.1 x 10(5); 1.4 x 10(5); and 9.2 x 10(5); 5.7 x 10(5); 1.2 x 10(5) to zero (0.0) in T6 = 8 h, respectively on the transparent and opaque toothbrushes. The reduction in viable microorganisms was more obvious with the transparent toothbrushes, although the number of viable microorganisms was not significantly different for the two types of toothbrushes at the end of the experiment, T5 = 1.4 x 10(5) (transparent) and T5 = 1.2 x 10(5) (opaque). CONCLUSIONS: With both opaque and transparent toothbrushes, the number of microorganisms decreased with time. A reduction in the number of microorganisms on the transparent toothbrushes was observed following inoculation and incubation. This suggests the transparent toothbrushes inhibit the viability of the S. mutans.

Effect of air humidity on the egg viability of predatory mites (Acari: Phytoseiidae, Stigmaeidae) common on rubber trees in Brazil

Rubber pest mites, Calacarus heveae and Tenuipalpus heveae, reach economic damage levels at the end of the rainy season and the beginning of the dry season in Brazil. Therefore, low humidity adaptation might be an important characteristic for predatory mites to successfully control pest organisms. This study determined the effect of the relative humidity (RH) levels of 30-100% on the hatching of larvae of Amblyseius acalyphus, Euseius citrifolius, Iphiseiodes zuluagai, Metaseiulus camelliae, Agistemus floridanus and Zetzellia malvinae at 25 ± 0.5°C. These predatory mites are common on rubber trees in the state of São Paulo and might be used for introduction in the major rubber tree production regions in the state of Mato Grosso. At 70% RH or higher, viability was 70% or higher for all species, indicating that their performance might be higher during the rainy season than during the dry season. Eggs of E. citrifolius and M. camelliae presented higher viability at the lower relative humidity levels than those of other species, indicating that these species might have higher chance to persist in the dry season. It is suggested that M. camelliae should be further evaluated for introduction in the state of Mato Grosso, considering that this mite is not yet present in that area. © Springer 2006.

Developed and studies on technical and economical viability of an environmental efficient urinal

This work introduces an innovative urinal for public convenience, that promotes at the same time water reuse and personal higiene, in a safe and economical way . Furthermore it demonstrates the latest technology and its technical and economical viabillity of utilization in new and already existing buildings facilities. This new model of personal higiene equipment offers as main benefits the improved economy with subsequent decrease in drinkable water consumption, sanitary safety, low cost and easy installation due to its simplicity and to the fact that it can be installed in already existing facilities. The proposal is constituted by a higienic, ecological and smart system for flushing of public urinals. It is a conjugated system of lavatory and urinal that reuses hands higienization water from the lavatory for flushing purpose. The proposed urinal can be operated manually or automatically by means of a presential sensor. The system promotes drinkable water economy by a rational utilization by avoiding the use of waste water from hand washing in place of clean water for flushing. The proposed equipment increases the economy of clean water in a simple and economical way and it can be installed in any type of public lavatory facilitie such as schools, public buildings, hospitals, commercial buildings, bus terminals, airports, stadiums, parking buildings and shopping centers. Additional benefits of the proposed system is the suggestion of hands washing before and after the use of the urinal without contamination risks from focet handling.and render more attractive the installation for a rational use of clean water in commercial and industrial buildings. Pay-back has shown to be very attractive for a number of internal return rates and also very attractive from the point of view of environmental protection.

Evaluation of tissue reaction, cell viability and cytokine production induced by Sealapex Plus

The aim of this study was to investigate the effects of mineral trioxide aggregate (MTA), Sealapex, and a combination of Sealapex and MTA (Sealapex Plus) on the reaction of subcutaneous connective tissue of rats, and on cell viability and cytokine production in mouse fibroblasts. The tissue reaction was carried out with dentin tubes containing the materials implanted in the dorsal connective tissue of rats. The histological analysis was performed after 7 and 30 days. Millipore culture plate inserts with polyethylene tubes filled with materials were placed into 24-well cell culture plates with mouse fibroblasts to evaluate the cell viability by MTT assay. ELISA assays were also performed after 24 h of exposure of the mouse fibroblasts to set material disks. Histopathologic examination showed Von Kossa-positive granules that were birefringent to polarized light for all the studied materials at the tube openings. No material inhibited the cell viability in the in vitro test. It was detected IL-6 production in all root-end filling materials. MTA and Sealapex Plus induced a slight raise of mean levels of IL-1β. The results suggest that Sealapex Plus is biocompatible and stimulates the mineralization of the tissue.

Effects of drying rate and storage time on Magnolia ovata Spreng. seed viability

Magnolia ovata seeds have been reported as desiccation sensitive. In order to test if the drying rate would affect the assessment of storage behaviour of these seeds, the effect of different drying rates and storage times on the viability was tested. Seeds were dried over activated silica gel (fast drying) or salt solutions for different periods (slow drying) and stored at -20°C. Partial drying transiently increased the final germination and the germination speed index, but further drying resulted in reduction of these parameters. Drying rate affected the final germination and vigour. Seeds that were slow-dried to 0.10 g H 2O ̇ g -1 dw retained high viability when compared with seeds desiccated to the same water content level by the fast drying method, although their vigour was reduced. Only slow-dried seeds could be stored at -20°C for 90 d without reduction of viability. These data suggested that the storage behaviour of seeds of M. ovata seeds should be classified as intermediate.

Effectiveness of mechanical brushing with different denture cleansing agents in reducing in vitro Candida albicans biofilm viability

The adhesion of Candida albicans to surfaces is the prerequisite for occurrence of denture stomatitis, a common disease diagnosed among denture wearers. A routine of denture cleansing is essential to prevent biofilm formation and the onset of this infection. The aim of this study was to investigate the effectiveness of combining brushing and cleansing agents in killing C. albicans biofilm. Disks of acrylic resin were made, sterilized, and inoculated with C. albicans (107 cfu/mL). After incubation (37°C/48 h), specimens were randomly assigned to 10 experimental groups (n=9): 5 subjected to brushing with distilled water or cleansing agents - dentifrice slurry, 2% chlorhexidine gluconate (CHX), 1% sodium hypochlorite (NaOCl), and Polident fresh cleanse® (combined method) - and 4 exposed to the cleansing agents without brushing (immersion). Non-cleansed specimens were used as positive controls. The viability of cells was evaluated by XTT reduction method. Results were analyzed by Mann-Whitney and Kruskal-Wallis tests (α=0.05). The combined method was significantly more effective (p<0.0001) in reducing biofilm viability than the immersion. Brushing with CHX and NaOCl resulted in 100% removal of the biofilm. Immersion in the agents reduced significantly (p<0.0001) the biofilm viability, with CHX being the most effective (p<0.0001). The use of the combined method of brushing with cleansing agents is an effective method to reduce C. albicans biofilm, being CHX and NaOCl the most effective solutions.

A hydrogel scaffold that maintains viability and supports differentiation of dental pulp stem cells

Objectives: The clinical translation of stem cell-based Regenerative Endodontics demands further development of suitable injectable scaffolds. Puramatrix™ is a defined, self-assembling peptide hydrogel which instantaneously polymerizes under normal physiological conditions. Here, we assessed the compatibility of Puramatrix™ with dental pulp stem cell (DPSC) growth and differentiation. Methods: DPSC cells were grown in 0.05-0.25% Puramatrix™. Cell viability was measured colorimetrically using the WST-1 assay. Cell morphology was observed in 3D modeling using confocal microscopy. In addition, we used the human tooth slice model with Puramatrix™ to verify DPSC differentiation into odontoblast-like cells, as measured by expression of DSPP and DMP-1. Results: DPSC survived and proliferated in Puramatrix™ for at least three weeks in culture. Confocal microscopy revealed that cells seeded in Puramatrix™ presented morphological features of healthy cells, and some cells exhibited cytoplasmic elongations. Notably, after 21 days in tooth slices containing Puramatrix™, DPSC cells expressed DMP-1 and DSPP, putative markers of odontoblastic differentiation. Significance: Collectively, these data suggest that self-assembling peptide hydrogels might be useful injectable scaffolds for stem cell-based Regenerative Endodontics. © 2012 Academy of Dental Materials.

Effect of Removing Seminal Plasma Using a Sperm Filter on the Viability of Refrigerated Stallion Semen

Cooling of equine semen obtained from some stallions results in lower seminal quality and viability when the seminal plasma (SP) is present. The objective of this study was to evaluate the effect of the removal of SP using a Sperm Filter on the viability of cooled stallion semen. For this purpose, 31 stallions were used. Their ejaculates were divided into three groups: CN, semen was diluted with an extender; FLT, SP was removed by filtration; and CT, SP was removed by centrifugation and cooled to 15°C for 24 hours. Sperm kinetics and plasma membrane integrity were evaluated immediately after collection (T0) and after 24 hours of refrigeration (T1). No difference (P > .05) was noted at T1 for total sperm motility (TM), progressive sperm motility, or plasma membrane integrity when semen samples from all the stallions were analyzed. However, when samples from stallions termed bad coolers were analyzed (TM = <30% at T1), a difference was observed in TM and progressive sperm motility for CN compared with FLT and CT at T1. Sperm recovery was greater when SP was removed using the filter (FLT) to that when the SP was removed by centrifugation (CN) (89% vs. 81%). Thus, we concluded that filtering with a Sperm Filter is an efficient and practical method for removal of SP from stallion ejaculates, with lower sperm loss than centrifugation. We also found that the presence of SP reduces the quality and viability of cooled semen from stallions whose semen is sensitive to the process of refrigeration. © 2013 Elsevier Inc.