Functional transfer of Salmonella pathogenicity island 2 to Salmonella bongori and Escherichia coli.

The type III secretion system (T3SS) encoded by the Salmonella pathogenicity island 2 (SPI2) has a central role in systemic infections by Salmonella enterica and for the intracellular phenotype. Intracellular S. enterica uses the SPI2-encoded T3SS to translocate a set of effector proteins into the host cell, which modify host cell functions, enabling intracellular survival and replication of the bacteria. We sought to determine whether specific functions of the SPI2-encoded T3SS can be transferred to heterologous hosts Salmonella bongori and Escherichia coli Mutaflor, species that lack the SPI2 locus and loci encoding effector proteins. The SPI2 virulence locus was cloned and functionally expressed in S. bongori and E. coli. Here, we demonstrate that S. bongori harboring the SPI2 locus is capable of secretion of SPI2 substrate proteins under culture conditions, as well as of translocation of effector proteins under intracellular conditions. An SPI2-mediated cellular phenotype was induced by S. bongori harboring the SPI2 if the sifA locus was cotransferred. An interference with the host cell microtubule cytoskeleton, a novel SPI2-dependent phenotype, was observed in epithelial cells infected with S. bongori harboring SPI2 without additional effector genes. S. bongori harboring SPI2 showed increased intracellular persistence in a cell culture model, but SPI2 transfer was not sufficient to confer to S. bongori systemic pathogenicity in a murine model. Transfer of SPI2 to heterologous hosts offers a new tool for the study of SPI2 functions and the phenotypes of individual effectors.

Salmonella pathogenicity island 2 mediates protection of intracellular Salmonella from reactive nitrogen intermediates.

Salmonella typhimurium causes an invasive disease in mice that has similarities to human typhoid. A type III protein secretion system encoded by Salmonella pathogenicity island 2 (SPI2) is essential for virulence in mice, as well as survival and multiplication within macrophages. Reactive nitrogen intermediates (RNI) synthesized by inducible nitric oxide synthase (iNOS) are involved in the control of intracellular pathogens, including S. typhimurium. We studied the effect of Salmonella infection on iNOS activity in macrophages. Immunofluorescence microscopy demonstrated efficient colocalization of iNOS with bacteria deficient in SPI2 but not wild-type Salmonella, and suggests that the SPI2 system interferes with the localization of iNOS and Salmonella. Furthermore, localization of nitrotyrosine residues in the proximity was observed for SPI2 mutant strains but not wild-type Salmonella, indicating that peroxynitrite, a potent antimicrobial compound, is excluded from Salmonella-containing vacuoles by action of SPI2. Altered colocalization of iNOS with intracellular Salmonella required the function of the SPI2-encoded type III secretion system, but not of an individual "Salmonella translocated effector." Inhibition of iNOS increased intracellular proliferation of SPI2 mutant bacteria and, to a lesser extent, of wild-type Salmonella. The defect in systemic infection of a SPI2 mutant strain was partially restored in iNOS(-/-) mice. In addition to various strategies to detoxify RNI or repair damage due to RNI, avoidance of colocalization with RNI is important in adaptation of a pathogen to an intracellular life style.

Shrewd alliances: mixed foraging associations between treeshrews,greater racket-tailed drongos and sparrowhawks on Great Nicobar Island, India

Mixed-species foraging associations may form to enhance feeding success or to avoid predators. We report the costs and consequences of an unusual foraging association between an endemic foliage gleaning tupaid (Nicobar treeshrew Tupaia nicobarica) and two species of birds; one an insectivorous commensal (greater racket-tailed drongo Dicrurus paradiseus) and the other a diurnal raptor and potential predator (Accipiter sp.). In an alliance driven, and perhaps engineered, by drongos, these species formed cohesive groups with predictable relationships. Treeshrew breeding pairs were found more frequently than solitary individuals with sparrowhawks and were more likely to tolerate sparrowhawks in the presence of drongos. Treeshrews maintained greater distances from sparrowhawks than drongos, and permitted the raptors to come closer when drongos were present. Treeshrew foraging rates declined in the presence of drongos; however, the latter may provide them predator avoidance benefits. The choice of the raptor to join the association is intriguing; particular environmental resource states may drive the evolution of such behavioural strategies. Although foraging benefits seem to be the primary driver of this association, predator avoidance also influences interactions, suggesting that strategies driving the formation of flocks may be complex and context dependent with varying benefits for different actors.

Effectors of Salmonella pathogenicity island 2: An island crucial to the life of Salmonella

The tug of war between a pathogen and its host has been one of the most amazing stories in the field of microbial pathogenesis for ages. The strongest known species of all living organisms is the Homo sapiens and yet it is incredible how a pathogen of the size of few microns is smart enough to defeat this mightiest group of survivors. It is of utmost interest to understand the mechanisms behind the successful habitation of a pathogen inside the ever-resisting and complicate human body. Numerous examples of diseases caused by such pathogens exist which intrigues us to venture in the world of host-pathogen interactions.

Salmonella methylglyoxal detoxification by STM3117-encoded lactoylglutathione lyase affects virulence in coordination with Salmonella pathogenicity island 2 and phagosomal acidification

Intracellular pathogens such as Salmonella enterica serovar Typhimurium (S. Typhimurium) manipulate their host cells through the interplay of various virulence factors. A multitude of such virulence factors are encoded on the genome of S. Typhimurium and are usually organized in pathogenicity islands. The virulence-associated genomic stretch of STM3117-3120 has structural features of pathogenicity islands and is present exclusively in non-typhoidal serovars of Salmonella. It encodes metabolic enzymes predicted to be involved in methylglyoxal metabolism. STM3117-encoded lactoylglutathione lyase significantly impacts the proliferation of intracellular Salmonella. The deletion mutant of STM3117 (Delta lgl) fails to grow in epithelial cells but hyper-replicates in macrophages. This difference in proliferation outcome was the consequence of failure to detoxify methylglyoxal by Delta lgl, which was also reflected in the form of oxidative DNA damage and upregulation of kefB in the mutant. Within macrophages, the toxicity of methylglyoxal adducts elicits the potassium efflux channel (KefB) in the mutant which subsequently modulates the acidification of mutant-containing vacuoles (MCVs). The perturbation in the pH of the MCV milieu and bacterial cytosol enhances the Salmonella pathogenicity island 2 translocation in Delta lgl, increasing its net growth within macrophages. In epithelial cells, however, the maturation of Delta lgl-containing vacuoles were affected as these non-phagocytic cells maintain less acidic vacuoles compared to those in macrophages. Remarkably, ectopic expression of Toll-like receptors 2 and 4 on epithelial cells partially restored the survival of Delta lgl. This study identified a novel metabolic enzyme in S. Typhimurium whose activity during intracellular infection within a given host cell type differentially affected the virulence of the bacteria.

Photographic analysis of natural and impounded salt marsh in the vicinity of Merritt Island, Florida

Qualitative analyses of available photographs and maps of Merritt Island, Florida provide a large-scale, historical perspective of ecological changes of the marshes in the vicinity. Sites that deserve closer scrutiny can be identified. Secondarily, such an analysis provides a geographical orientation essential for communication not only between newcomers and those familiar with the area, but also among those familiar with the area but who refer to sites by differing methods. Photographs and maps from various sources were examined. Below are listed what we consider to be the most useful subset of these for ecological and geographical assessment of salt marsh impoundments on Merritt Island, Florida. (Document has 25 pages.)

Categorized bibliography for a conceptual model of salt marsh management on Merritt Island, Florida

Enclosed is a bibliography of 556 published articles, technical reports, theses, dissertations, and books that form the basis for a conceptual model of salt marsh management on Merritt Island, Florida (Section 1). A copy of each item is available on file at the Florida Cooperative Fish and Wildlife Research Unit, Gainesville. Some relevant proprietary items and unpublished drafts have not been included pending permission of the authors. We will continue to add pertinent references to our bibliography and files. Currently, some topics are represented by very few items. As our synthesis develops, we will be able to indicate a subset of papers most pertinent to an understanding of the ecology and management of Merritt Island salt marshes. (98 page document)

Conceptual model of salt marsh management on Merritt Island National Wildlife Refuge, Florida: final report

Diking and holding water on salt marshes ("impounding" the marsh) is a management technique used on Merritt Island National Wildlife Refuge (MINWR) and elsewhere in the Southeast to: a) prevent the reproduction of saltmarsh mosquitos, and b) attract wintertering waterfowl and other marsh, shore, and wading birds. Because of concern that diking and holding water may interfere with the production of estuarine fish and shellfish, impoundment managers are being asked to consider altering management protocol to reduce or eliminate any such negative influence. How to change protocol and preserve effective mosquito control and wildlife management is a decision of great complexity because: a) the relationships between estuarine organisms and the fringing salt marshes at the land-water interface are complex, and b) impounded marshes are currently good habitat for a variety of species of fish and wildlife. Most data collection by scientists and managers in the area has not been focused on this particular problem. Furthermore, collection of needed data may not be possible before changes in protocol are demanded. Therefore, the purpose of this document is two-fold: 1) to suggest management alternatives, given existing information, and 2) to help identify research needs that have a high probability of leading to improved simultaneous management of mosquitos, waterfowl, other wildlife, freshwater fish, and estuarine fish and shellfish on the marshland of the Merritt Island National Wildlife Refuge. (92 page document)

Ichthyoplankton of the continental shelf near Kodiak Island, Alaska

Eleven ichthyoplankton surveys were conducted (1 in 1972 and 10 between 1977 and 1979) in the northeastern Pacific Ocean over the continental shelf off Kodiak Island, Alaska. In the 677 neuston and 632 bongo tows, eggs or larvae of more than 80 fish taxa were found. They were present in every season and throughout the survey area, although more taxa and more individuals were found in summer than in other seasons. Among the more abundant species were the gadid Theragra chalcogramma and several hexagrammids and pleuronectids. The hexagrammids and several coUids were abundant in the neustonic layer, where they spent close to a year as larvae and prejuvenlles. Although the seasonal and geographic distribution of most taxa was complex, two patterns emerged: Late summer-fall spawners produce demersal eggs and have neustonic larvae that remain pelagic for several months (hexagrammids and some cottlds), and spring-summer spawners have pelagic eggs and larvae that spend several weeks in the plankton but are not closely associated with the surface (Theragra chalcogramma, pleuronectlds). (PDF file contains 95 pages.)

Measures of turbidity in coastal waters, Coconut Island, Oahu, Hawaii, August 31 - September 2, 2005: workshop proceedings

A three day workshop on turbidity measurements was held at the Hawaii Institute of Marine Biology from August 3 1 to September 2, 2005. The workshop was attended by 30 participants from industry, coastal management agencies, and academic institutions. All groups recognized common issues regarding the definition of turbidity, limitations of consistent calibration, and the large variety of instrumentation that nominally measure "turbidity." The major recommendations, in order of importance for the coastal monitoring community are listed below: 1. The community of users in coastal ecosystems should tighten instrument design configurations to minimize inter-instrument variability, choosing a set of specifications that are best suited for coastal waters. The IS0 7027 design standard is not tight enough. Advice on these design criteria should be solicited through the ASTM as well as Federal and State regulatory agencies representing the majority of turbidity sensor end users. Parties interested in making turbidity measurements in coastal waters should develop design specifications for these water types rather than relying on design standards made for the analysis of drinking water. 2. The coastal observing groups should assemble a community database relating output of specific sensors to different environmental parameters, so that the entire community of users can benefit from shared information. This would include an unbiased, parallel study of different turbidity sensors, employing a variety of designs and configuration in the broadest range of coastal environments. 3. Turbidity should be used as a measure of relative change in water quality rather than an absolute measure of water quality. Thus, this is a recommendation for managers to develop their own local calibrations. See next recommendation. 4. If the end user specifically wants to use a turbidity sensor to measure a specific water quality parameter such as suspended particle concentration, then direct measurement of that water quality parameter is necessary to correlate with 'turbidity1 for a particular environment. These correlations, however, will be specific to the environment in which they are measured. This works because there are many environments in which water composition is relatively stable but varies in magnitude or concentration. (pdf contains 22 pages)