Effects of gaseous atmosphere and antioxidants on the development and cryotolerance of bovine embryos at different periods of in vitro culture

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

Opportunistic microorganisms in patients with head and neck trauma

The use of antimicrobial agents has facilitated the dissemination of multi-resistant microorganisms, compromising dental and medical treatment. The aim of this study was to evaluate the distribution of different opportunistic microbial species in patients who suffered head and neck trauma, under temporary maintenance in nosocomial environment, particularly intensive care units, on the occurrence of such microorganisms in the oral cavity of the patients. It was selected 38 patients subjected to head and neck traumas. After emergency surgical procedures, clinical samples of saliva, sub and supragingival biofilms and mucosal surfaces were collected at two different moments: just after stabilization of the patient and soon after patients’ release from medical units. The presence of opportunistic and superinfecting microorganisms was evaluated by culture on selective and non-selective media, and the presence of the family Enterobacteriaceae, as well as genera Enterococcus, Pseudomonas, and Staphylococcus was assessed by PCR. It was found that the use of antimicrobials, even for short periods of time was sufficient to facilitate colonization by microorganisms of the families Enterobacteriaceae and Pseudomonadaceae, as well as yeasts and enterococci. These results support the concept that medical and dental teams should make a periodically change of antimicrobials used in treatment protocols in hospital for head and neck trauma patients, in order to minimize dissemination of opportunistic or superinfecting microorganisms.

Bamboo as sustainable material used in design and civil construction: species, management, characterization and applications

The use of bamboo as construction and raw material for producing products can be considered a feasible alternative to the abusive use of steel, concrete and oil byproducts. Its use can also reduce the pressure on the use of wood from native and planted forests. Although there are thousands of bamboo species spread about the world and Brazil itself has hundreds of native species, the use and basic knowledge of its characteristics and applications are still little known and little disseminated. This paper's main objective is to introduce the species, the management phases, the physical and mechanical characteristics and the experiences in using bamboo in design and civil construction as per the Bamboo Project implemented at UNESP, Bauru campus since 1994. The results are divided into: a) Field activities - description of the technological species of interest, production chain flows, types of preservative treatments and clump management practices for the development, adaptation and production of different species of culms; b) Lab experiments - physical and mechanical characterization of culms processed as laminated strips and as composite material (glue laminated bamboo – glubam); c) Uses in projects - experiences with natural bamboo and glubam in design, architecture and civil construction projects. In the final remarks, the study aims to demonstrate, through practical and laboratory results, the material's multi-functionality and the feasibility in using bamboo as a sustainable material.

Animal-side serologic assay for rapid detection of Mycobacterium Bovis infection in multiple species of free-ranging wildlife

Numerous species of mammals are susceptible to Mycobacterium bovis, the causative agent of bovine tuberculosis (TB). Several wildlife hosts have emerged as reservoirs of M. bovis infection for domestic livestock in different countries. In the present study, blood samples were collected from Eurasian badgers (n = 1532), white-tailed deer (n = 463), brushtail possums (n = 129), and wild boar (n = 177) for evaluation of antibody responses to M. bovis infection by a lateral-flow rapid test (RT) and multiantigen print immunoassay (MAPIA). Magnitude of the antibody responses and antigen recognition patterns varied among the animals as determined by MAPIA; however, MPB83 was the most commonly recognized antigen for each host studied. Other seroreactive antigens included ESAT-6, CFP10, and MPB70. The agreement of the RT with culture results varied from 74% for possums to 81% for badgers to 90% for wild boar to 97% for white-tailed deer. Small numbers of wild boar and deer exposed to M. avium infection or paratuberculosis, respectively, did not cross-react in the RT, supporting the high specificity of the assay. In deer, whole blood samples reacted similarly to corresponding serum specimens (97% concordance), demonstrating the potential for field application. As previously demonstrated for badgers and deer, antibody responses to M. bovis infection in wild boar were positively associated with advanced disease. Together, these findings suggest that a rapid TB assay such as the RT may provide a useful screening tool for certain wildlife species that may be implicated in the maintenance and transmission of M. bovis infection to domestic livestock.

Comparison of Gross Pathology, Histopathology, and Mycobacterial Culture for the Diagnosis of Tuberculosis in Elk (Cervus elaphus)

Using the isolation of Mycobacterium bovis as the reference standard, this study evaluated the sensitivity, specificity and kappa statistic of gross pathology (abattoir postmortem inspection), histopathology, and parallel or series combinations of the two for the diagnosis of tuberculosis in 430 elk and red deer. Two histopathology interpretations were evaluated: histopathology I, where the presence of lesions compatible with tuberculosis was considered positive, and histopathology II, where lesions compatible with tuberculosis or a select group of additional possible diagnoses were considered positive. In the 73 animals from which M. bovis was isolated, gross lesions of tuberculosis were most often in the lung (48), the retropharyngeal lymph nodes (36), the mesenteric lymph nodes (35), and the mediastinal lymph nodes (16). Other mycobacterial isolates included: 11 M. paratuberculosis, 11 M. avium, and 28 rapidly growing species or M. terrae complex. The sensitivity estimates of gross pathology and histopathology I were 93% (95% confidence limits [CL] 84,97%) and 88% [CL 77,94%], respectively, and the specificity of both was 89% [CL 85,92%]). The sensitivity and specificity of histopathology II were 89% (CL 79,95%) and 77% (CL 72,81%), respectively. The highest sensitivity estimates (93- 95% [CL 84,98%]) were obtained by interpreting gross pathology and histopathology in parallel (where an animal had to be positive on at least one of the two, to be classified as combination positive). The highest specificity estimates (94-95% [CL 91-97%]) were generated when the two tests were interpreted in series (an animal had to be positive on both tests to be classified as combination positive). The presence of gross or microscopic lesions showed moderate to good agreement with the isolation of M. bovis (Kappa = 65-69%). The results show that post-mortem inspection, histopathology and culture do not necessarily recognize the same infected animals and that the spectra of animals identified by the tests overlaps.

Community structure and dynamics of Monogenea and Trematoda in three North American cyprinid species in the Salt Valley Watershed, Nebraska.

An investigation was made of the communities of gill monogene genus Dactylogyrus (Platyhelminthes, Monogenea) and the populations of blackspot parasite (Platyhelminthes, Trematoda) of Pimephales promelas, Notropis stramineus, and Semotilus atromaculatus in 3 distinct sites along the 3 converging tributaries in southeastern Nebraska from 2004 to 2006. This work constitutes the first multi-site, multi-year study of a complex community of Dactylogyrus spp. and their reproductive activities on native North American cyprinid species. The biological hypothesis that closely related species with direct lifecycles respond differently to shared environmental conditions was tested. It was revealed that in this system that, Cyprinid species do not share Dactylogyrus species, host size and sex are not predictive of infection, and Dactylogyrus community structure is stable, despite variation in seasonal occurrence and populations among sites. The biological hypothesis that closely related species have innate differences in reproductive activities that provide structure to their populations and influence their roles in the parasite community was tested. It was revealed that in this system, host size, sex, and collection site are not predictive of reproductive activities, that egg production is not always continuous and varies in duration among congeners, and that recruitment of larval Dactylogyrus is not continuous across parasites’ reproductive periods. Hatch timing and host availability, not reproductive timing, are the critical factors determining population dynamics of the gill monogenes in time and space. Lastly, the biological hypothesis that innate blackspot biology is responsible for parasite host-specificity, host recruitment strategies and parasite population structure was tested. Field collections revealed that for blackspot, host size, sex, and collection month and year are not predictive of infection, that parasite cysts survive winter, and that host movement is restricted among the 3 collection sites. Finally, experimental infections of hosts with cercaria isolated from 1st intermediate snail hosts reveal that cercarial biology, not environmental circumstances, are responsible for differences in infection among hosts.

Detection Methods for the Genus Lysobacter and the Species Lysobacter enzymogenes

Strains of Lysobacter enzymogenes, a bacterial species with biocontrol activity, have been detected via 16S rDNA sequences in soil in different parts of the world. In most instances, however, their occurrence could not be confirmed by isolation, presumably because the species occurred in low numbers relative to faster-growing species of Bacillus or Pseudomonas. In this study, we developed DNA-based detection and enrichment culturing methods for Lysobacter spp. and L. enzymogenes specifically. In the DNA-based method, a region of 16S rDNA conserved among Lysobacter spp. (L4: GAG CCG ACG TCG GAT TAG CTA GTT), was used as the forward primer in PCR amplification. When L4 and universal bacterial primer 1525R were used to amplify DNA from various bacterial species, an 1100-bp product was found in Lysobacter spp. exclusively. The enrichment culturing method involved culturing soils for 3 days in a chitin-containing broth amended with antibiotics. Bacterial strains in the enrichment culture were isolated on yeast-cell agar and then identified by 16S rDNA sequence analysis. A strain of L. enzymogenes added to soils was detected at populations as low as 102 and 104 CFU/g soil by PCR amplification and enrichment culturing, respectively. In a survey of 58 soil samples, Lysobacter was detected in 41 samples by PCR and enrichment culture, out of which 6 yielded strains of Lysobacter spp. by enrichment culture. Among isolated strains, all were identified to be L. enzymogenes, with the exception of a strain of L. antibioticus. Although neither method alone is completely effective at detecting L. enzymogenes, they are complementary when used together and may provide new information on the spatial distribution of the species in soil.

Identification of multi-component trail pheromones in the most evolutionarily derived termites, the Nasutitermitinae (Termitidae)

In the present study, trail pheromone blends are identified for the first time in termites. In the phylogenetically complex Nasutitermitinae, trail-following pheromones are composed of dodecatrienol and neocembrene, the proportions of which vary according to species, although neocembrene is always more abundant than dodecatrienol (by 25-250-fold). Depending on species, termites were more sensitive to dodecatrienol or to neocembrene but the association of both components always elicited significantly higher trail following, with a clear synergistic effect in most of the studied species. A third component, trinervitatriene, was identified in the sternal gland secretion of several species, but its function remains unknown. The secretion of trail pheromone blends appears to be an important step in the evolution of chemical communication in termites. The pheromone optimizes foraging, and promotes their ecological success. (C) 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 99, 20-27.

Bacterial leakage along the implant-abutment interface: culture and DNA Checkerboard hybridization analyses

Objective Bacterial species have been found harboring the internal surface of dental implants as consequence of their failed connections. The aim of the present study was to compare the detection frequency of bacterial leakage from human saliva through the implantabutment interface, under non-loading conditions, using either DNA Checkerboard or culture method. Materials and methods Thirty dental implants with hexagonal platforms were connected to pre-machined abutments according to the manufacturers specifications. The assemblies were individually incubated in human saliva under anaerobic conditions for 7 similar to days at 37 degrees C. Afterward, contents from the inner parts of the implants were collected and evaluated with either DNA Checkerboard (s similar to=similar to 15) or culture (n similar to=similar to 15). Subsequently, identification and quantitation of bacterial species from saliva and implants were carried out for the group evaluated with the DNA Checkerboard method. Results Both DNA Checkerboard and culture showed positive signals of bacterial leakage in 6 of the 15 evaluated samples. Capnocytophaga gingivalis and Streptococcus mutans were the most frequently detected species harboring the internal surface of the implants followed by Veillonella parvula. Conclusion Occurrence of bacterial leakage along the implantabutment interface is comparably detected with both DNA Checkerboard hybridization and conventional culture methods.

Semiotics of Communication: From Semiosis of Nature to Culture

Communication Studies currently undergoes a crisis of paradigms that requires an ontological review that must begin with a debate about the conditions of possibility of every communicational phenomena. In this article we argue that semiosis offers a conceptual framework that allows for the study of communication as qualitative action. Semiosis, or the action of the sign, is here defined as a fundamental process based on perception that models the world of species, creating cognition and culture. At the core of semiosis are dynamic structures that the authors have defined as 'ontological diagrams'. The first purpose of Semiotics of Communication is to understand how these modeling systems evolve ontologically and phylogenically, producing, in the case of human culture, means of communication ever more varied and technologically advanced.

Mapping the biosphere: exploring species to understand the origin, organization and sustainability of biodiversity

The time is ripe for a comprehensive mission to explore and document Earth's species. This calls for a campaign to educate and inspire the next generation of professional and citizen species explorers, investments in cyber-infrastructure and collections to meet the unique needs of the producers and consumers of taxonomic information, and the formation and coordination of a multi-institutional, international, transdisciplinary community of researchers, scholars and engineers with the shared objective of creating a comprehensive inventory of species and detailed map of the biosphere. We conclude that an ambitious goal to describe 10 million species in less than 50 years is attainable based on the strength of 250 years of progress, worldwide collections, existing experts, technological innovation and collaborative teamwork. Existing digitization projects are overcoming obstacles of the past, facilitating collaboration and mobilizing literature, data, images and specimens through cyber technologies. Charting the biosphere is enormously complex, yet necessary expertise can be found through partnerships with engineers, information scientists, sociologists, ecologists, climate scientists, conservation biologists, industrial project managers and taxon specialists, from agrostologists to zoophytologists. Benefits to society of the proposed mission would be profound, immediate and enduring, from detection of early responses of flora and fauna to climate change to opening access to evolutionary designs for solutions to countless practical problems. The impacts on the biodiversity, environmental and evolutionary sciences would be transformative, from ecosystem models calibrated in detail to comprehensive understanding of the origin and evolution of life over its 3.8 billion year history. The resultant cyber-enabled taxonomy, or cybertaxonomy, would open access to biodiversity data to developing nations, assure access to reliable data about species, and change how scientists and citizens alike access, use and think about biological diversity information.

Repertoire, Genealogy and Genomic Organization of Cruzipain and Homologous Genes in Trypanosoma cruzi, T. cruzi-Like and Other Trypanosome Species

Trypanosoma cruzi, the agent of Chagas disease, is a complex of genetically diverse isolates highly phylogenetically related to T. cruzi-like species, Trypanosoma cruzi marinkellei and Trypanosoma dionisii, all sharing morphology of blood and culture forms and development within cells. However, they differ in hosts, vectors and pathogenicity: T. cruzi is a human pathogen infective to virtually all mammals whilst the other two species are non-pathogenic and bat restricted. Previous studies suggest that variations in expression levels and genetic diversity of cruzipain, the major isoform of cathepsin L-like (CATL) enzymes of T. cruzi, correlate with levels of cellular invasion, differentiation, virulence and pathogenicity of distinct strains. In this study, we compared 80 sequences of genes encoding cruzipain from 25 T. cruzi isolates representative of all discrete typing units (DTUs TcI-TcVI) and the new genotype Tcbat and 10 sequences of homologous genes from other species. The catalytic domain repertoires diverged according to DTUs and trypanosome species. Relatively homogeneous sequences are found within and among isolates of the same DTU except TcV and TcVI, which displayed sequences unique or identical to those of TcII and TcIII, supporting their origin from the hybridization between these two DTUs. In network genealogies, sequences from T. cruzi clustered tightly together and closer to T. c. marinkellei than to T. dionisii and largely differed from homologues of T. rangeli and T. b. brucei. Here, analysis of isolates representative of the overall biological and genetic diversity of T. cruzi and closest T. cruzi-like species evidenced DTU- and species-specific polymorphisms corroborating phylogenetic relationships inferred with other genes. Comparison of both phylogenetically close and distant trypanosomes is valuable to understand host-parasite interactions, virulence and pathogenicity. Our findings corroborate cruzipain as valuable target for drugs, vaccine, diagnostic and genotyping approaches.

Evolutionary Insights from Bat Trypanosomes: Morphological, Developmental and Phylogenetic Evidence of a New Species, Trypanosoma (Schizotrypanum) erneyi sp nov., in African Bats Closely Related to Trypanosoma (Schizotrypanum) cruzi and Allied Species

Parasites of the genus Trypanosoma are common in bats and those of the subgenus Schizotrypanum are restricted to bats throughout the world, with the exception of Trypanosoma (Schizotrypanum) cruzi that also infects other mammals and is restricted to the American Continent. We have characterized trypanosome isolates from Molossidae bats captured in Mozambique, Africa. Morphology and behaviour in culture, supported by phylogenetic inferences using SSU (small subunit) rRNA, gGAPDH (glycosomal glyceraldehyde 3-phosphate dehydrogenase) and Cyt b (cytochrome b) genes, allowed to classify the isolates as a new Schizotrypanum species named Trypanosoma (Schizotrypanum) erneyi sp. nov. This is the first report of a Schizotrypanum species from African bats cultured, characterized morphologically and biologically, and positioned in phylogenetic trees. The unprecedented finding of a new species of the subgenus Schizotrypanum from Africa that is closest related to the America-restricted Trypanosoma (Schizotrypanum) cruzi marinkellei and T. cruzi provides new insights into the origin and evolutionary history of T. cruzi and closely related bat trypanosomes. Altogether, data from our study support the hypothesis of an ancestor trypanosome parasite of bats evolving to infect other mammals, even humans, and adapted to transmission by triatomine bugs in the evolutionary history of T. cruzi in the New World. (c) 2012 Elsevier GmbH. All rights reserved.

Multi-strange baryon production in pp collisions at root s=7 TeV with ALICE

A measurement of the multi-strange Xi(-) and Omega(-) baryons and their antiparticles by the ALICE experiment at the CERN Large Hadron Collider (LHC) is presented for inelastic proton-proton collisions at a centre-of-mass energy of 7 TeV. The transverse momentum (p(T)) distributions were studied at mid-rapidity (vertical bar y vertical bar < 0.5) in the range of 0.6 < p(T) < 8.5 GeV/c Xi(-) for and Xi(+) baryons, and in the range of 0.8 < P-T < 5 GeV/c for Omega(-) and<(Omega)over bar>(+). Baryons and antibaryons were measured as separate particles and we find that the baryon to antibaryon ratio of both particle species is consistent with unity over the entire range of the measurement. The statistical precision of the current data has allowed us to measure a difference between the mean p(T) of Xi(-) ((Xi) over bar)(+) and Omega(-) ((Omega) over bar (+)). Particle yields, mean pi, and the spectra in the intermediate pi range are not well described by the PYTHIA Perugia 2011 tune Monte Carlo event generator, which has been tuned to reproduce the early LHC data. The discrepancy is largest for Omega(-)((Omega) over bar (+)). This PYTHIA tune approaches the pi spectra of Xi(-) and Xi(+) baryons below p(T) <0.85 GeV/c and describes the Xi(-) and Xi(+) spectra above p(T) > 6.0 GeV/c. We also illustrate the difference between the experimental data and model by comparing the corresponding ratios of (Omega(-) +(Omega) over bar (+))/(Xi(-) + Xi(+)) as a function of transverse mass. (C) 2012 CERN. Published by Elsevier B.V. All rights reserved.

Trypanosoma (Megatrypanum) melophagium in the Sheep Ked Melophagus ovinus from Organic Farms in Croatia: Phylogenetic Inferences Support Restriction to Sheep and Sheep Keds and Close Relationship with Trypanosomes from Other Ruminant Species

Trypanosoma (Megatrypanum) melophagium is a parasite of sheep transmitted by sheep keds, the sheep-restricted ectoparasite Melophagus ovinus (Diptera: Hippoboscidae). Sheep keds were 100% prevalent in sheep from five organic farms in Croatia, Southeastern Europe, whereas trypanosomes morphologically compatible with T. melophagium were 86% prevalent in the guts of the sheep keds. Multilocus phylogenetic analyses using sequences of small subunit rRNA, glycosomal glyceraldehyde-3-phosphate dehydrogenase, spliced leader, and internal transcribed spacer 1 of the rDNA distinguished T. melophagium from all allied trypanosomes from other ruminant species and placed the trypanosome in the subgenus Megatrypanum. Trypanosomes from sheep keds from Croatia and Scotland, the only available isolates for comparison, shared identical sequences. All biologic and phylogenetic inferences support the restriction of T. melophagium to sheep and, especially, to the sheep keds. The comparison of trypanosomes from sheep, cattle, and deer from the same country, which was never achieved before this work, strongly supported the host-restricted specificity of trypanosomes of the subgenus Megatrypanum. Our findings indicate that with the expansion of organic farms, both sheep keds and T. melophagium may re-emerge as parasitic infections of sheep.