Molecular characterization of virulence factors in Aeromonas hydrophila obtained from fish

Multiple factors can be involved in the virulence processes of Aeromonas hydrophila. The objective of the present paper was to verify the presence of aerolysin, hidrolipase, elastase and lipase virulence genes through the polymerase chain reaction (PCR) in A. hydrophila isolates obtained from fish of the São Francisco River Valley, and to evaluate virulence according to the presence of these genes in Nile tilapia fingerlings. One hundred and fourteen isolates from the bacteria were used. DNA was heat extracted and PCR undertaken using specific primers described in the literature. For in vivo tests Nile tilapia fingerlings were used. From the PCR tests, negative isolates for all genes tested were selected, positive isolates for two genes (aerolysin and elastase) and positive for the four genes tested. These were inoculated at a concentration of 10(8) UFC/ml into the tilapias, considered as treatments; another group of animals was used as control (with inoculation of saline solution). In all, 12 distinct standards regarding the presence of virulence factors in isolates from A. hydrophila, were observed. Of the 114 isolates analyzed, 100 (87.72%) presented at least one of the virulence factors under study. The virulence factors were widely distributed among the A. hydrophila isolates. Aerolysin was the most frequent virulence factor present in the isolates analyzed. A. hydrophila led to the mortality of the Nile tilapia fingerlings, regardless of the absence or quantity of virulence genes tested.

A field survey of crop management practices and distribution of ALS resistant Euphorbia heterophylla in two states in southern Brazil

Herbicide resistance was reported in Brazil almost ten years ago. One of the main weeds with herbicide resistance is wild poinsettia (Euphorbia heterophylla). This work evaluates the distribution of ALS-resistant E. heterophylla in two states in southern Brazil and determines the major contributing management causes for weed resistance selection in the area. E. heterophylla seeds from 148 sites located in Paraná and Rio Grande do Sul were sampled during 2001 and 2002. Farmers provided specific site data for weed control, tillage system, crop rotation and harvesting operations during previous years. ALS resistant E. heterophylla biotypes were found widely distributed in the survey area. Data analysis suggests seed dissemination is unlikely to explain the widespread distribution of resistance. The most probable factor for the selection of the resistant E. heterophylla is the persistent high use of ALS-inhibiting herbicides over time. Indirect evidence is presented demonstrating the need to educate legislators and farmers about the importance of herbicide mixtures as a strategy to prevent herbicide resistance.

Effect of light and temperature on germination and early growth of Vochysia tucanorum Mart., Vochysiaceae, in cerrado and forest soil under different radiation levels

Vochysia tucanorum Mart. (Vochysiaceae) is a very widely distributed species in the cerrado and forests in Brazil, from the State of Bahia to Paraná. Its seed germination was analysed under eight treatments using different conditions of light and temperature. Up to 73 days, at least ca. 93% of the seeds germinated in all treatments, except under alternating temperatures of 35°/10°C (62%). Light enhanced germination, except under constant temperature of 25°C when seeds are not photoblastic. At this temperature, the highest percentage of germination was found. Initial vegetative growth was analysed by comparing the effects of shading, by means of different percentages of full daylight (100%, 45%, 10.6%, 4.8%), and soils from cerrado and forest. During 123 days, no mortality was recorded and cotyledons persisted on the seedlings of all treatments. In general, the seedlings of V. tucanorum were found to be more or less indifferent to the soil from cerrado or forest and shading seems to promote height increment. However, total dry mass was larger under a radiation of 45% full daylight and smaller under extreme conditions (100% or 4.8% full daylight).

Reproductive biology of the herkogamous vine Chiococca alba (L.) Hitchc. (Rubiaceae) in the Atlantic Rain Forest, SE Brazil

The family Rubiaceae comprises a wide spectrum of floral mechanisms and two of them seem to be common in certain groups, e.g., distyly in Rubioidae and styllar pollen in Ixoroidae. These mechanisms include herkogamy, which is interpreted as a strategy that avoids self-pollination. This is the first report on the reproductive biology of Chiococca alba, a species that is widely distributed in America. We studied floral biology and the mating system, which were evaluated through fruit set comparisons after controlled crosses (self- and cross-pollinations and test for apomixis), as well as through the evaluation of pollen tube growth resulting from these controlled crosses. Flowers of C. alba are herkogamous, cream, protandrous and lasted for two days. No measurable nectar was found, despite the presence of a nectary-like structure at the base of the corolla tube. Chiococca alba is a preferentially self-incompatible species, but self-pollination and apomixis also contribute to the natural fruit-set. Its reproductive strategy (herkogamy associated with protandry) is different from that expected for members of Chiococceae tribe (i.e., styllar pollen presentation).

Subgeneric diversity of Brasilonema (Cyanobacteria, Scytonemataceae)

The recently described scytonematoid cyanobacterial genus Brasilonema is known mainly from tropical and subtropical rain forests (Mata Atlântica) of southeastern Brazil, where it occurs in aerophytic wooden, stony and iron substrates. This genus was defined according to both molecular and morphological criteria. The type species B. bromeliae was described from the specialized habitat: it grows in phytothelmes, epiphytic on both living and died leaves within the rosettes of large bromeliad plants slightly above or in the zone of the water level. The genus Brasilonema is evidently widely distributed in coastal forests of São Paulo State, where it occurs also in remarkable diversity. According to our results, this genus currently comprises seven taxa, which are distinct by different morphology and ecological characteristics.

Test of hypotheses about herbivory and chemical defences of Qualea parviflora (Vochysiaceae) in Brazilian Cerrado

(Test of hypotheses about herbivory and chemical defences of Qualea parviflora (Vochysiaceae) in Brazilian Cerrado). Qualea parviflora Mart. (Vochysiaceae), a widely distributed tree found in different habitats in Brazilian Cerrado (savanna), provides resources for a great variety of insects. In this study, we tested two hypotheses about plant investment in anti-herbivore defences along a fertility gradient in Cerrado: the carbon/nitrogen balance (CNBH) and resource availability (RAH). We also investigated how the pattern of herbivory varies through the year and among three types of vegetation in Brazilian Cerrado - campo sujo, cerrado sensu strictu and cerradão. Sampling was conducted in three types of vegetation and in rainy (January and November) and dry months (April and July). Damage on 20 completely expanded leaves, leaf nutrients, sclerophlylly, total phenols and tannins were recorded for each plant (n = 30). When leaves were young, less sclerophyllous, and with higher concentration of nutrients and tannins, damage by herbivores was about 7% in cerrado sensu stricto and 3% in campo sujo. Mature leaves did not show any significant difference on herbivory among habitats, that varied from 6 to 9%. Nutrient availability to plants is an important factor determining production of secondary metabolites in Q. parviflora, corroborating the CNBH. The absence of correlation between damaged leaf area and tannin concentration did not corroborate the RAH, suggesting that tannin production is not strongly influenced by herbivores on Q. parviflora.

The genera Chara and Nitella (Chlorophyta, Characeae) in the subtropical Itaipu Reservoir, Brazil

The family Characeae, represented by two genera in Brazil, Chara and Nitella, is considered to include the closest living relatives of land plants, and its members play important ecological role in aquatic ecosystems. The present taxonomic survey of Chara and Nitella was performed in tributaries that join to form the Brazilian shore of the Itaipu Reservoir on the Paraná River. Thirteen species were recorded, illustrated, and described: C. braunii var. brasiliensis R.Bicudo, C. guairensis R.Bicudo, N. acuminata A.Braun ex Wallman, N. furcata (Roxburgh ex Bruzileus) C.Agardh, and N. subglomerata A.Braun, already cited for the reservoir, and C. hydropitys Reichenbach, C. rusbyana Howe, N. axillaris A.Braun, N. glaziovii G.Zeller, N. gracilis (Smith) C.Agardh, N. hyalina (DC.) C.Agardh, N. inversa Imahori, and N. microcarpa A.Braun that represent new occurrences for the Itaipu Reservoir and Paraná State. Among the species encountered, C. guairensis, N. furcata, and N. glaziovii are widely distributed, while C. hydropitys and C. rusbyana have more restricted distributions.

Distribution of the a2, a3, and a5 nicotinic acetylcholine receptor subunits in the chick brain

Nicotinic acetylcholine receptors (nAChRs) are ionotropic receptors comprised of a and ß subunits. These receptors are widely distributed in the central nervous system, and previous studies have revealed specific patterns of localization for some nAChR subunits in the vertebrate brain. In the present study we used immunohistochemical methods and monoclonal antibodies to localize the a2, a3, and a5 nAChR subunits in the chick mesencephalon and diencephalon. We observed a differential distribution of these three subunits in the chick brain, and showed that the somata and neuropil of many central structures contain the a5 nAChR subunit. The a2 and a3 subunits, on the other hand, exhibited a more restricted distribution than a5 and other subunits previously studied, namely a7, a8 and ß2. The patterns of distribution of the different nAChR subunits suggest that neurons in many brain structures may contain several subtypes of nAChRs and that in a few regions one particular subtype may determine the cholinergic nicotinic responses

Distribution of calcium-binding proteins in the chick visual system

The calcium-binding proteins calbindin (CB), calretinin (CR), and parvalbumin (PV) have been extensively studied over the last decade since they appear to be important as buffers of intracellular calcium. In the present study we investigated the distribution of these proteins in the chick visual system by means of conventional immunocytochemistry. The results indicated that CB, CR, and PV are widely distributed in retinorecipient areas of the chick brain. In some regions, all three calcium-binding proteins were present at different intensities and often in different neurons such as in the dorsolateral thalamic complex. In other areas, such as the nucleus geniculatus lateralis ventralis, only CB and CR were detected, whereas PV was absent. These results show that these three calcium-binding proteins are differentially distributed in the visual system of the chick, with varying degrees of co-localization

Galectins: a key intersection between glycobiology and immunology

Galectins are a family of evolutionarily conserved animal lectins, widely distributed from lower invertebrates to mammals. They share sequence and structure similarities in the carbohydrate recognition domain and specificity for polylactosamine-enriched glycoconjugates. In the last few years significant experimental data have been accumulated concerning their participation in different biological processes requiring carbohydrate recognition such as cell adhesion, cell growth regulation, inflammation, immunomodulation, apoptosis and metastasis. In the present review we will discuss some exciting questions and advances in galectin research, highlighting the significance of these proteins in immunological processes and their implications in biomedical research, disease diagnosis and clinical intervention. Designing novel therapeutic strategies based on carbohydrate recognition will provide answers for the treatment of autoimmune disorders, inflammatory processes, allergic reactions and tumor spreading.

Pollen mother cells of Tradescantia clone 4430 and Tradescantia pallida var. purpurea are equally sensitive to the clastogenic effects of X-rays

The Tradescantia micronucleus test is a sensitive bioassay for mutagenesis that may be employed both under field and laboratory conditions. This test has been standardized mostly on the basis of the results obtained with clone 4430. However, this clone is not well adapted to tropical weather, frequently showing problems with growth and flowering. In addition, it is attacked by parasites and insects, a fact that limits its use in field studies aiming at the biomonitoring of air pollution. In the city of São Paulo, Tradescantia pallida (Rose) Hunt. var. purpurea Boom is widely distributed as an ornamental plant in gardens and along roadsides and streets, mostly because of its natural resistance and its easy propagation. In this report, we present dose-response curves indicating that the sensitivity of T. pallida and clone 4430 to X-radiation (1, 10, 25 and 50 cGy) is similar. The results confirm our previous suggestion that T. pallida represents a good alternative for in situ mutagenesis testing in tropical regions, especially biomonitoring studies in which the exposure conditions may not be fully controllable.

Proteinase activity regulation by glycosaminoglycans

There are few reports concerning the biological role and the mechanisms of interaction between proteinases and carbohydrates other than those involved in clotting. It has been shown that the interplay of enzymes and glycosaminoglycans is able to modulate the activity of different proteases and also to affect their structures. From the large number of proteases belonging to the well-known protease families and also the variety of carbohydrates described as widely distributed, only few events have been analyzed more deeply. The term "family" is used to describe a group of proteases in which every member shows an evolutionary relationship to at least one other protease. This relationship may be evident throughout the entire sequence, or at least in that part of the sequence responsible for catalytic activity. The majority of proteases belong to the serine, cysteine, aspartic or metalloprotease families. By considering the existing limited proteolysis process, in addition to the initial idea that the proteinases participate only in digestive processes, it is possible to conclude that the function of the enzymes is strictly limited to the cleavage of intended substrates since the destruction of functional proteins would result in normal tissue damage. In addition, the location as well as the eventual regulation of protease activity promoted by glycosaminoglycans can play an essential role in the development of several physiopathological conditions.

Physiological implications of the regulation of vacuolar H+-ATPase by chloride ions

Vacuolar H+-ATPase is a large multi-subunit protein that mediates ATP-driven vectorial H+ transport across the membranes. It is widely distributed and present in virtually all eukaryotic cells in intracellular membranes or in the plasma membrane of specialized cells. In subcellular organelles, ATPase is responsible for the acidification of the vesicular interior, which requires an intraorganellar acidic pH to maintain optimal enzyme activity. Control of vacuolar H+-ATPase depends on the potential difference across the membrane in which the proton ATPase is inserted. Since the transport performed by H+-ATPase is electrogenic, translocation of H+-ions across the membranes by the pump creates a lumen-positive voltage in the absence of a neutralizing current, generating an electrochemical potential gradient that limits the activity of H+-ATPase. In many intracellular organelles and cell plasma membranes, this potential difference established by the ATPase gradient is normally dissipated by a parallel and passive Cl- movement, which provides an electric shunt compensating for the positive charge transferred by the pump. The underlying mechanisms for the differences in the requirement for chloride by different tissues have not yet been adequately identified, and there is still some controversy as to the molecular identity of the associated Cl--conducting proteins. Several candidates have been identified: the ClC family members, which may or may not mediate nCl-/H+ exchange, and the cystic fibrosis transmembrane conductance regulator. In this review, we discuss some tissues where the association between H+-ATPase and chloride channels has been demonstrated and plays a relevant physiologic role.

Mitochondrial control region haplotypes of the South American sea lion Otaria flavescens (Shaw, 1800)

The South American sea lion, Otaria flavescens, is widely distributed along the Pacific and Atlantic coasts of South America. However, along the Brazilian coast, there are only two nonbreeding sites for the species (Refúgio de Vida Silvestre da Ilha dos Lobos and Refúgio de Vida Silvestre do Molhe Leste da Barra do Rio Grande), both in Southern Brazil. In this region, the species is continuously under the effect of anthropic activities, mainly those related to environmental contamination with organic and inorganic chemicals and fishery interactions. This paper reports, for the first time, the genetic diversity of O. flavescens found along the Southern Brazilian coast. A 287-bp fragment of the mitochondrial DNA control region (D-loop) was analyzed. Seven novel haplotypes were found in 56 individuals (OFA1-OFA7), with OFA1 being the most frequent (47.54%). Nucleotide diversity was moderate (π = 0.62%) and haplotype diversity was relatively low (67%). Furthermore, the median joining network analysis indicated that Brazilian haplotypes formed a reciprocal monophyletic clade when compared to the haplotypes from the Peruvian population on the Pacific coast. These two populations do not share haplotypes and may have become isolated some time back. Further genetic studies covering the entire species distribution are necessary to better understand the biological implications of the results reported here for the management and conservation of South American sea lions.

Modulatory effects of taurine on jejunal contractility

Taurine (2-aminoethanesulfonic acid) is widely distributed in animal tissues and has diverse pharmacological effects. However, the role of taurine in modulating smooth muscle contractility is still controversial. We propose that taurine (5-80 mM) can exert bidirectional modulation on the contractility of isolated rat jejunal segments. Different low and high contractile states were induced in isolated jejunal segments of rats to observe the effects of taurine and the associated mechanisms. Taurine induced stimulatory effects on the contractility of isolated rat jejunal segments at 3 different low contractile states, and inhibitory effects at 3 different high contractile states. Bidirectional modulation was not observed in the presence of verapamil or tetrodotoxin, suggesting that taurine-induced bidirectional modulation is Ca2+dependent and requires the presence of the enteric nervous system. The stimulatory effects of taurine on the contractility of isolated jejunal segments was blocked by atropine but not by diphenhydramine or by cimetidine, suggesting that muscarinic-linked activation was involved in the stimulatory effects when isolated jejunal segments were in a low contractile state. The inhibitory effects of taurine on the contractility of isolated jejunal segments were blocked by propranolol and L-NG-nitroarginine but not by phentolamine, suggesting that adrenergic β receptors and a nitric oxide relaxing mechanism were involved when isolated jejunal segments were in high contractile states. No bidirectional effects of taurine on myosin phosphorylation were observed. The contractile states of jejunal segments determine taurine-induced stimulatory or inhibitory effects, which are associated with muscarinic receptors and adrenergic β receptors, and a nitric oxide associated relaxing mechanism.