Morphology of the midgut of Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae) adult ticks in different feeding stages

The intestinal epithelial cells of ticks are fundamental for their full feeding and reproductive success, besides being considered important sites for the development of pathogens. Rhipicephalus sanguineus ticks are known for their great medical and veterinary importance, and for this reason, the knowledge of their intestinal morphology may provide relevant subsidies for the control of these animals, either by direct acaricidal action over these cells or by the production of vaccines. Therefore, this study aimed to describe the midgut morphology of male and female R. sanguineus ticks in different feeding stages, by means of histological analysis. Significant differences were observed between the genders, and such alterations may refer mainly to the distinct demands for nutrients, much higher in females, which need to develop and carry out the egg-laying process. In general, the midgut is coated by a thin muscle layer and presents a pseudostratified epithelium, in which two basic types of cells can be observed, connected to a basal membrane - generative or stem and digestive cells. The latter was classified as follows: residual, deriving from the phase anterior to ecdysis; pinocytic, with vesicles containing liquid or pre-digested components of blood; phagocytic, with entire cells or remnants of nuclear material inside cytoplasmic vesicles; and mature, free in the lumen. Digestion is presumably intracellular and asynchronous and corresponds to a process which starts with the differentiation of generative cells into pinocytic digestive cells, which subsequently start to phagocytize intact blood cells and finally detach from the epithelium, being eliminated with feces. © 2012 Springer-Verlag Berlin Heidelberg.

First record of chilodonella hexasticha (ciliophora: Chilodonellidae) in brazilian cultured fish: A morphological and pathological assessment

Chilodonelids are small ciliated protozoans found worldwide and can be dangerous in culture conditions. This study presents morphometric data on the ciliate Chilodonella that is found in cultured Nile tilapia (Oreochromis niloticus), native bait fish tuvira (Gymnotus aff. inaequilabiatus) and native pacu (Piaractus mesopotamicus) and includes a histopathological assessment of the changes that occur in the pacu. For parasitic diagnosis, skin and gill samples were scraped onto slides, dried at room temperature, stained with Giemsa or impregnated with silver nitrate, and the measurements were obtained from photomicrographs. In the diseased pacu, the first gill arch was collected and fixed in a 10% buffered formalin solution for histopathological analysis. Parasite specimens from the different collection sites were identified morphologically as C. hexasticha Kiernik (1909). Diseased fish exhibited depigmentation, skin ulceration, scale loss, excessive mucus production and gill lesions. Histopathological analysis of pacu gills displayed epithelial proliferation with mononuclear inflammatory infiltrate, hemorrhages, and scattering necrosis. In Brazilian-farmed fish this is the first record of C. hexasticha, which has great pathogenic potential in cultured freshwater species. In addition, two new hosts are presented. © 2012 Elsevier B.V.

Filamentous fungi from the Atlantic marine sponge Dragmacidon reticulatum

Dragmacidon reticulatum is a marine sponge of wide occurrence in the Eastern and Western Atlantic. Little is known about D. reticulatum fungal diversity. Filamentous fungi recovered from D. reticulatum were assessed in the present study using a polyphasic taxonomic approach, including classical morphology, molecular biology and MALDI-TOF ICMS. Ninety-eight fungal strains were isolated from two D. reticulatum samples by using six different culture media, which were identified up to the genus level. Sixty-four distinct fungal ribotypes were obtained, distributed among twenty-four different genera belonging to the Ascomycota and Zygomycota. Representatives of Penicillium and Trichoderma were the most diverse and abundant fungi isolated. Amongst Penicillium spp. three isolates belonged to the same ribotype can be considered as a putative new species. Data derived from the present study highlight the importance of using a polyphasic approach to get an accurate identification in order to structure a reliable culture collection. © 2012 Springer-Verlag Berlin Heidelberg.

Neurotoxic action of permethrin in Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae) female ticks. Morphological and cytochemical evaluation of the central nervous system

Studies on the molecular bases of the neurotoxic action of acaricides are found in the literature; but there are no studies of this action on the nervous system of ticks at the cellular level. The present study describes the morphological and cytochemical changes in the synganglion of Rhipicephalus sanguineus semi-engorged females exposed to different concentrations of permethrin, a pyrethroid with recognized neurotoxic action. Permethrin induced the degeneration of the synganglion through a process of apoptosis involving autophagy, characterized by the condensation and margination of the chromatin, formation of blebs in the nuclear envelope and fragmentation of the nucleus, loss of shape of neural cells and integrity of cellular membrane, cytoplasmic shrinkage, and lower levels of acid phosphatase in the nervous tissue as the concentration of permethrin increased. This study provided further evidence of the neurotoxic action of permethrin, which impairs the metabolism of R. sanguineus nervous systems, and consequently the physiology of other systems, dependent on the neural control. These results provide cytochemical and histological confirmation of the neurotoxic action of permethrin, previously inferred from molecular and tick behavioral evidence. © 2013 Elsevier B.V.

You and I: The Effects of Intra- and Inter-Species Interactions on Microbial Biofilms, Growth, and Morphology

Humanity is shaped by its relationships with microbes. From bacterial infections to the production of biofuels, industry and health often hinge on our control of microbial populations. Understanding the physiological and genetic basis of their behaviors is therefore of the highest importance. To this end I have investigated the genetic basis of plastic adhesion in Saccharomyces cerevisiae, the mechanistic and evolutionary dynamics of mixed species biofilms with Escherichia coli and S. cerevisiae, and the induction of filamentation in E. coli. Using a bulk segregant analysis on experimentally evolved populations, I detected 28 genes that are likely to mediate plastic adhesion in S. cerevisiae. With a variety of imaging and culture manipulation techniques, I found that particular strains of E. coli are capable of inducing flocculation and macroscopic biofilm formation via coaggregation with yeast. I also employed experimental evolution and microbial demography techniques to find that selection for mixed species biofilm association leads to lower fecundity in S. cerevisiae. Using culture manipulation and imaging techniques, I also found that E. coli are capable of inducing a filamentous phenotype with a secreted signal that has many of the qualities of a quorum sensing molecule.

Systematics, functional morphology and distribution of a bivalve (Apachecorbula muriatica gen. et sp. nov.) from the rim of the ‘Valdivia Deep’ brine pool in the Red Sea

The deep brine pools of the Red Sea comprise extreme, inhospitable habitats yet house microbial communities that potentially may fuel adjacent fauna. We here describe a novel bivalve from a deep-sea (1525 m) brine pool in the Red Sea, where conditions of high salinity, lowered pH, partial anoxia and high temperatures are prevalent. Remotely operated vehicle (ROV) footage showed that the bivalves were present in a narrow (20 cm) band along the rim of the brine pool, suggesting that it is not only tolerant of such extreme conditions but is also limited to them. The bivalve is a member of the Corbulidae and named Apachecorbula muriatica gen. et sp. nov. The shell is atypical of the family in being modioliform and thin. The semi-infaunal habit is seen in ROV images and reflected in the anatomy by the lack of siphons. The ctenidia are large and typical of a suspension feeding bivalve, but the absence of guard cilia and the greatly reduced labial palps suggest that it is non-selective as a response to low food availability. It is proposed that the low body mass observed is a consequence of the extreme habitat and low food availability. It is postulated that the observed morphology of Apachecorbula is a result of paedomorphosis driven by the effects of the extreme environment on growth but is in part mitigated by the absence of high predation pressures.

Simulation of stromatolite growth using diffusion-limited aggregation and a discrete model of biogeochemical exchanges in microbial mat

Resume : Mieux comprendre les stromatolithes et les tapis microbiens est un sujet important en biogéosciences puisque cela aide à l'étude des premières formes de vie sur Terre, a mieux cerner l'écologie des communautés microbiennes et la contribution des microorganismes a la biominéralisation, et même à poser certains fondements dans les recherches en exobiologie. D'autre part, la modélisation est un outil puissant utilisé dans les sciences naturelles pour appréhender différents phénomènes de façon théorique. Les modèles sont généralement construits sur un système d'équations différentielles et les résultats sont obtenus en résolvant ce système. Les logiciels disponibles pour implémenter les modèles incluent les logiciels mathématiques et les logiciels généraux de simulation. L'objectif principal de cette thèse est de développer des modèles et des logiciels pour aider a comprendre, via la simulation, le fonctionnement des stromatolithes et des tapis microbiens. Ces logiciels ont été développés en C++ en ne partant d'aucun pré-requis de façon a privilégier performance et flexibilité maximales. Cette démarche permet de construire des modèles bien plus spécifiques et plus appropriés aux phénomènes a modéliser. Premièrement, nous avons étudié la croissance et la morphologie des stromatolithes. Nous avons construit un modèle tridimensionnel fondé sur l'agrégation par diffusion limitée. Le modèle a été implémenté en deux applications C++: un moteur de simulation capable d'exécuter un batch de simulations et de produire des fichiers de résultats, et un outil de visualisation qui permet d'analyser les résultats en trois dimensions. Après avoir vérifié que ce modèle peut en effet reproduire la croissance et la morphologie de plusieurs types de stromatolithes, nous avons introduit un processus de sédimentation comme facteur externe. Ceci nous a mené a des résultats intéressants, et permis de soutenir l'hypothèse que la morphologie des stromatolithes pourrait être le résultat de facteurs externes autant que de facteurs internes. Ceci est important car la classification des stromatolithes est généralement fondée sur leur morphologie, imposant que la forme d'un stromatolithe est dépendante de facteurs internes uniquement (c'est-à-dire les tapis microbiens). Les résultats avancés dans ce mémoire contredisent donc ces assertions communément admises. Ensuite, nous avons décidé de mener des recherches plus en profondeur sur les aspects fonctionnels des tapis microbiens. Nous avons construit un modèle bidimensionnel de réaction-diffusion fondé sur la simulation discrète. Ce modèle a été implémenté dans une application C++ qui permet de paramétrer et exécuter des simulations. Nous avons ensuite pu comparer les résultats de simulation avec des données du monde réel et vérifier que le modèle peut en effet imiter le comportement de certains tapis microbiens. Ainsi, nous avons pu émettre et vérifier des hypothèses sur le fonctionnement de certains tapis microbiens pour nous aider à mieux en comprendre certains aspects, comme la dynamique des éléments, en particulier le soufre et l'oxygène. En conclusion, ce travail a abouti à l'écriture de logiciels dédiés à la simulation de tapis microbiens d'un point de vue tant morphologique que fonctionnel, suivant deux approches différentes, l'une holistique, l'autre plus analytique. Ces logiciels sont gratuits et diffusés sous licence GPL (General Public License). Abstract : Better understanding of stromatolites and microbial mats is an important topic in biogeosciences as it helps studying the early forms of life on Earth, provides clues re- garding the ecology of microbial ecosystems and their contribution to biomineralization, and gives basis to a new science, exobiology. On the other hand, modelling is a powerful tool used in natural sciences for the theoretical approach of various phenomena. Models are usually built on a system of differential equations and results are obtained by solving that system. Available software to implement models includes mathematical solvers and general simulation software. The main objective of this thesis is to develop models and software able to help to understand the functioning of stromatolites and microbial mats. Software was developed in C++ from scratch for maximum performance and flexibility. This allows to build models much more specific to a phenomenon rather than general software. First, we studied stromatolite growth and morphology. We built a three-dimensional model based on diffusion-limited aggregation. The model was implemented in two C++ applications: a simulator engine, which can run a batch of simulations and produce result files, and a Visualization tool, which allows results to be analysed in three dimensions. After verifying that our model can indeed reproduce the growth and morphology of several types of stromatolites, we introduced a sedimentation process as an external factor. This lead to interesting results, and allowed to emit the hypothesis that stromatolite morphology may be the result of external factors as much as internal factors. This is important as stromatolite classification is usually based on their morphology, imposing that a stromatolite shape is dependant on internal factors only (i.e. the microbial mat). This statement is contradicted by our findings, Second, we decided to investigate deeper the functioning of microbial mats, We built a two-dimensional reaction-diffusion model based on discrete simulation, The model was implemented in a C++ application that allows setting and running simulations. We could then compare simulation results with real world data and verify that our model can indeed mimic the behaviour of some microbial mats. Thus, we have proposed and verified hypotheses regarding microbial mats functioning in order to help to better understand them, e.g. the cycle of some elements such as oxygen or sulfur. ln conclusion, this PhD provides a simulation software, dealing with two different approaches. This software is free and available under a GPL licence.

Assessing the microbial oxidative stress mechanism of ozone treatment through the responses of Escherichia coli mutants

Aims: To investigate the effect of the oxidative stress of ozone on the microbial inactivation, cell membrane integrity and permeability and morphology changes of Escherichia coli. Methods and Results: Escherichia coli BW 25113 and its isogenic mutants in soxR, soxS, oxyR, rpoS and dnaK genes were treated with ozone at a concentration of 6 lg ml)1 for a period up to 240 s. A significant effect of ozone exposure on microbial inactivation was observed. After ozonation, minor effects on the cell membrane integrity and permeability were observed, while scanning electron microscopy analysis showed slightly altered cell surface structure. Conclusions: The results of this study suggest that cell lysis was not the major mechanism of microbial inactivation. The deletion of oxidative stress–related genes resulted in increased susceptibility of E. coli cells to ozone treatment, implying that they play an important role for protection against the radicals produced by ozone. However, DnaK that has previously been shown to protect against oxidative stress did not protect against ozone treatment in this study. Furthermore, RpoS was important for the survival against ozone. Significance and Impact of the Study: This study provides important information about the role of oxidative stress in the responses of E. coli during ozonation.

Taxonomic Identity in Microbial Eukaryotes: A Practical Approach Using the Testate Amoeba Centropyxis to Resolve Conflicts Between Old and New Taxonomic Descriptions

The present work focuses on 12 taxa of the genus Centropyxis Stein, 1857 to explore the conflict between traditional and contemporary taxonomic practices. We examined the morphology, biometry, and ecology of 2,120 Centropyxis individuals collected from Tiete River, Sao Paulo, Brazil; with these new data we studied the consistency of previously described species, varieties, and forms. We encountered transitional forms of test morphology that undermine specific and varietal distinctions for three species and nine varieties. Biometrical analyses made comparing the organisms at the species level suggest a lack of separation between Centropyxis aculeata and Centropyxis discoides, and a possible distinction for Centropyxis ecornis based on spine characteristics. However, incongruence between recent and previous surveys makes taking any taxonomic-nomenclatural actions inadvisable, as they would only add to the confusion. We suggest an explicit and objective taxonomic practice in order to enhance our taxonomic and species concepts for microbial eukaryotes. This will allow more precise inferences of taxon identity for studies in other areas.

In vivo and in vitro evaluation of an Acetobacter xylinum synthesized microbial cellulose membrane intended for guided tissue repair

Background: Barrier materials as cellulose membranes are used for guided tissue repair. However, it is essential that the surrounding tissues accept the device. The present study histologically evaluated tissue reaction to a microbial cellulose membrane after subcutaneous implantation in mice. Furthermore, the interaction between mesenchymal stem cells and the biomaterial was studied in vitro to evaluate its ability to act as cellular scaffold for tissue engineering.Methods: Twenty-five Swiss Albino mice were used. A 10 x 10 mm cellulose membrane obtained through biosynthesis using Acetobacter xylinum bacteria was implanted into the lumbar subcutaneous tissue of each mouse. The mice were euthanatized at seven, 15, 30, 60, and 90 days, and the membrane and surrounding tissues were collected and examined by histology.Results: A mild inflammatory response without foreign body reaction was observed until 30 days post-surgery around the implanted membrane. Polarized microscopy revealed that the membrane remained intact at all evaluation points. Scanning electron microscopy of the cellulose membrane surface showed absence of pores. The in vitro evaluation of the interaction between cells and biomaterial was performed through viability staining analysis of the cells over the biomaterial, which showed that 95% of the mesenchymal stem cells aggregating to the cellulose membrane were alive and that 5% were necrotic. Scanning electron microscopy showed mesenchymal stem cells with normal morphology and attached to the cellulose membrane surface.Conclusion: The microbial cellulose membrane evaluated was found to be nonresorbable, induced a mild inflammatory response and may prove useful as a scaffold for mesenchymal stem cells.

In vitro effect of low-level laser therapy on typical oral microbial biofilms

The aim of this study was to evaluate the effect of specific parameters of low-level laser therapy (LLLT) on biofilms formed by Streptococcus mutans, Candida albicans or an association of both species. Single and dual-species biofilms - SSB and DSB - were exposed to laser doses of 5, 10 or 20 J/cm 2 from a near infrared InGaAsP diode laser prototype (LASERTable; 780 ± 3 nm, 0.04 W). After irradiation, the analysis of biobilm viability (MTT assay), biofilm growth (cfu/mL) and cell morphology (SEM) showed that LLLT reduced cell viability as well as the growth of biofilms. The response of S. mutans (SSB) to irradiation was similar for all laser doses and the biofilm growth was dose dependent. However, when associated with C. albicans (DSB), S. mutans was resistant to LLLT. For C. albicans, the association with S. mutans (DSB) caused a significant decrease in biofilm growth in a dose-dependent fashion. The morphology of the microorganisms in the SSB was not altered by LLLT, while the association of microbial species (DSB) promoted a reduction in the formation of C. albicans hyphae. LLLT had an inhibitory effect on the microorganisms, and this capacity can be altered according to the interactions between different microbial species.

In vitro evaluation of the microbial contamination on new toothbrushes: A preliminary study

Objective: The presence and survival of microorganisms on toothbrush bristles might play a role on the etiology of oral infections. The aim of this in vitro study was to evaluate the presence of bacterial contamination on new toothbrushes before oral contact. Materials and methods: Forty toothbrushes from five different manufacturers were used in this experimental study. Each manufacturer was divided according to conventional local of obtaining: industry, drugstore, market, and perfumery. The toothbrush heads were completely immersed into tubes containing 5.0 mL of sterile peptonated water (dilution 1:10). A group of eight tubes containing the sterile solution was used as control. After 21 days of anaerobic incubation, occurrence of contamination was visually evaluated and confirmed by light microscopy. Results: Bacterial growth in the medium, indicative of bristles contamination, was found in a total of 19 out of 40 samples (47.5%) evaluated: 6 out of 14 samples (42.85%) from industry group, 4 out of 8 samples (50.0%) from drugstore, 5 out of 10 samples (50.0%) from market, and 4 out of 8 samples (50.0%) from perfumery. Only the toothbrushes with bristles coated with chlorhexidine did not show contamination. The Gram-negative sporulating bacilli were the most prevalent form recovered. Conclusions: Except for chlorhexidine group, bacterial growth was observed in all groups evaluated irrespective local of obtaining. Microsc. Res. Tech., 2012. (c) 2011 Wiley Periodicals, Inc.

Prominent Effect of Soil Network Heterogeneity on Microbial Invasion

Using a network representation for real soil samples and mathematical models for microbial spread, we show that the structural heterogeneity of the soil habitat may have a very significant influence on the size of microbial invasions of the soil pore space. In particular, neglecting the soil structural heterogeneity may lead to a substantial underestimation of microbial invasion. Such effects are explained in terms of a crucial interplay between heterogeneity in microbial spread and heterogeneity in the topology of soil networks. The main influence of network topology on invasion is linked to the existence of long channels in soil networks that may act as bridges for transmission of microorganisms between distant parts of soil.

Relationship Between Orofacial Function, Dentofacial Morphology, And Bite Force In Young Subjects.

The aim was to evaluate the relationship between orofacial function, dentofacial morphology, and bite force in young subjects. Three hundred and sixteen subjects were divided according to dentition stage (early, intermediate, and late mixed and permanent dentition). Orofacial function was screened using the Nordic Orofacial Test-Screening (NOT-S). Orthodontic treatment need, bite force, lateral and frontal craniofacial dimensions and presence of sleep bruxism were also assessed. The results were submitted to descriptive statistics, normality and correlation tests, analysis of variance, and multiple linear regression to test the relationship between NOT-S scores and the studied independent variables. The variance of NOT-S scores between groups was not significant. The evaluation of the variables that significantly contributed to NOT-S scores variation showed that age and presence of bruxism related to higher NOT-S total scores, while the increase in overbite measurement and presence of closed lip posture related to lower scores. Bite force did not show a significant relationship with scores of orofacial dysfunction. No significant correlations between craniofacial dimensions and NOT-S scores were observed. Age and sleep bruxism were related to higher NOT-S scores, while the increase in overbite measurement and closed lip posture contributed to lower scores of orofacial dysfunction.