The Pseudomonas aeruginosa Magnesium Transporter MgtE Inhibits Transcription of the Type III Secretion System

Pseudomonas aeruginosa is an opportunistic pathogen that causes life-long pneumonia in individuals with cystic fibrosis (CF). These long-term infections are maintained by bacterial biofilm formation in the CF lung. We have recently developed a model of P. aeruginosa biofilm formation on cultured CF airway epithelial cells. Using this model, we discovered that mutation of a putative magnesium transporter gene, called mgtE, led to increased cytotoxicity of P. aeruginosa toward epithelial cells. This altered toxicity appeared to be dependent upon expression of the type III secretion system (T3SS). In this study, we found that mutation of mgtE results in increased T3SS gene transcription. Through epistasis analyses, we discovered that MgtE influences the ExsE-ExsC-ExsD-ExsA gene regulatory system of T3SS by either directly or indirectly inhibiting ExsA activity. While variations in calcium levels modulate T3SS gene expression in P. aeruginosa, we found that addition of exogenous magnesium did not inhibit T3SS activity. Furthermore, mgtE variants that were defective for magnesium transport could still complement the cytotoxicity effect. Thus, the magnesium transport function of MgtE does not fully explain the regulatory effects of MgtE on cytotoxicity. Overall, our results indicate that MgtE modulates expression of T3SS genes

Effects of commercial fishing regulations on stranding rates of bottlenose dolphin (Tursiops truncatus).

Fisheries management actions taken to protect one species can have unintended, and sometimes positive, consequences on other species. For example, regulatory measures to reduce fishing effort in the winter gillnet fishery for spiny dogfish (Squalus acanthias) off North Carolina (NC) also led to decreases in the number of bycaught bottlenose dolphins (Tursiops truncatus). This study found that a marked decrease in fishing effort for spiny dogfish in NC also corresponded with a marked decrease in winter stranding rates of bottlenose dolphins with entanglement lesions (P= 0.002). Furthermore, from 1997 through 2002, there was a significant positive correlation (r2 = 0.79; P= 0.0003) between seasonal bycatch estimates of bottlenose dolphins in gill nets and rates of stranded dolphins with entanglement lesions. With this information, stranding thresholds were developed that would enable the detection of those increases in bycatch in near real-time. This approach is valuable because updated bycatch estimates from observer data usually have a time-lag of two or more years. Threshold values could be used to detect increases in stranding rates, triggering managers immediately to direct observer effort to areas of potentially high bycatch or to institute mitigation measures. Thus, observer coverage and stranding investigations can be used in concert for more effective fishery management

Experiences with Establishment of a Multi-University Center of Academic Excellence in Information Assurance/Cyber Defense

The National Security Agency (NSA) and Department of Homeland Security (DHS), in response to an unmet workforce need for cybersecurity program graduates, jointly sponsor a program by which a post-secondary education institution may achieve recognition as a Center of Academic Excellence in Information Assurance/Cyber Defense (CAE IA/CD). The program identifies standards, criteria, and an evaluation process. Many individual institutions have achieved recognition. The University of Maine System, composed of seven universities, is the first multi-university entity to achieve the CAE IA/CD recognition. The purpose of this paper is to share the key challenges, opportunities, and experiences that contributed to this achievement, and offer recommendations

Bartonella species detection in captive, stranded and free-ranging cetaceans

We present prevalence of Bartonella spp. for multiple cohorts of wild and captive cetaceans. One hundred and six cetaceans including 86 bottlenose dolphins (71 free-ranging, 14 captive in a facility with a dolphin experiencing debility of unknown origin, 1 stranded), 11 striped dolphins, 4 harbor porpoises, 3 Risso's dolphins, 1 dwarf sperm whale and 1 pygmy sperm whale (all stranded) were sampled. Whole blood (n = 95 live animals) and tissues (n = 15 freshly dead animals) were screened by PCR (n = 106 animals), PCR of enrichment cultures (n = 50 animals), and subcultures (n = 50 animals). Bartonella spp. were detected from 17 cetaceans, including 12 by direct extraction PCR of blood or tissues, 6 by PCR of enrichment cultures, and 4 by subculture isolation. Bartonella spp. were more commonly detected from the captive (6/14, 43%) than from free-ranging (2/71, 2.8%) bottlenose dolphins, and were commonly detected from the stranded animals (9/21, 43%; 3/11 striped dolphins, 3/4 harbor porpoises, 2/3 Risso's dolphins, 1/1 pygmy sperm whale, 0/1 dwarf sperm whale, 0/1 bottlenose dolphin). Sequencing identified a Bartonella spp. most similar to B. henselae San Antonio 2 in eight cases (4 bottlenose dolphins, 2 striped dolphins, 2 harbor porpoises), B. henselae Houston 1 in three cases (2 Risso's dolphins, 1 harbor porpoise), and untyped in six cases (4 bottlenose dolphins, 1 striped dolphin, 1 pygmy sperm whale). Although disease causation has not been established, Bartonella species were detected more commonly from cetaceans that were overtly debilitated or were cohabiting in captivity with a debilitated animal than from free-ranging animals. The detection of Bartonella spp. from cetaceans may be of pathophysiological concern

Strandings as indicators of marine mammal biodiversity and human interactions off the coast of North Carolina

The adjacency of 2 marine biogeographic regions off Cape Hatteras, North Carolina (NC), and the proximity of the Gulf Stream result in a high biodiversity of species from northern and southern provinces and from coastal and pelagic habitats. We examined spatiotemporal patterns of marine mammal strandings and evidence of human interaction for these strandings along NC shorelines and evaluated whether the spatiotemporal patterns and species diversity of the stranded animals reflected published records of populations in NC waters. During the period of 1997–2008, 1847 stranded animals were documented from 1777 reported events. These animals represented 9 families and 34 species that ranged from tropical delphinids to pagophilic seals. This biodiversity is higher than levels observed in other regions. Most strandings were of coastal bottlenose dolphins (Tursiops truncatus) (56%), harbor porpoises (Phocoena phocoena) (14%), and harbor seals (Phoca vitulina) (4%). Overall, strandings of northern species peaked in spring. Bottlenose dolphin strandings peaked in spring and fall. Almost half of the strandings, including southern delphinids, occurred north of Cape Hatteras, on only 30% of NC’s coastline. Most stranded animals that were positive for human interaction showed evidence of having been entangled in fishing gear, particularly bottlenose dolphins, harbor porpoises, short-finned pilot whales (Globicephala macrorhynchus), harbor seals, and humpback whales (Megaptera novaeangliae). Spatiotemporal patterns of bottlenose dolphin strandings were similar to ocean gillnet fishing effort. Biodiversity of the animals stranded on the beaches reflected biodiversity in the waters off NC, albeit not always proportional to the relative abundance of species (e.g., Kogia species). Changes in the spatiotemporal patterns of strandings can serve as indicators of underlying changes due to anthropogenic or naturally occurring events in the source populations

Step-Wise Loss of Bacterial Flagellar Torsion Confers Progressive Phagocytic Evasion

Phagocytosis of bacteria by innate immune cells is a primary method of bacterial clearance during infection. However, the mechanisms by which the host cell recognizes bacteria and consequentially initiates phagocytosis are largely unclear. Previous studies of the bacterium Pseudomonas aeruginosa have indicated that bacterial flagella and flagellar motility play an important role in colonization of the host and, importantly, that loss of flagellar motility enables phagocytic evasion. Here we use molecular, cellular, and genetic methods to provide the first formal evidence that phagocytic cells recognize bacterial motility rather than flagella and initiate phagocytosis in response to this motility. We demonstrate that deletion of genes coding for the flagellar stator complex, which results in non-swimming bacteria that retain an initial flagellar structure, confers resistance to phagocytic binding and ingestion in several species of the gamma proteobacterial group of Gram-negative bacteria, indicative of a shared strategy for phagocytic evasion. Furthermore, we show for the first time that susceptibility to phagocytosis in swimming bacteria is proportional to mot gene function and, consequently, flagellar rotation since complementary genetically- and biochemically-modulated incremental decreases in flagellar motility result in corresponding and proportional phagocytic evasion. These findings identify that phagocytic cells respond to flagellar movement, which represents a novel mechanism for non-opsonized phagocytic recognition of pathogenic bacteria

Independent evolution of shape and motility allows evolutionary flexibility in Firmicutes bacteria

Functional morphological adaptation is an implicit assumption across many ecological studies. However, despite a few pioneering attempts to link bacterial form and function, functional morphology is largely unstudied in prokaryotes. One intriguing candidate for analysis is bacterial shape, as multiple lines of theory indicate that cell shape and motility should be strongly correlated. Here we present a large-scale use of modern phylogenetic comparative methods to explore this relationship across 325 species of the phylum Firmicutes. In contrast to clear predictions from theory, we show that cell shape and motility are not coupled, and that transitions to and from flagellar motility are common and strongly associated with lifestyle (free-living or host-associated). We find no association between shape and lifestyle, and contrary to recent evidence, no indication that shape is associated with pathogenicity. Our results suggest that the independent evolution of shape and motility in this group might allow a greater evolutionary flexibility

Monomethylarsonous Acid (MMAIII) Has an Adverse Effect on the Innate Immune Response of Human Bronchial Epithelial Cells to Pseudomonas Aeruginosa

Arsenic is the number one contaminant of concern with regard to human health according to the World Health Organization. Epidemiological studies on Asian and South American populations have linked arsenic exposure with an increased incidence of lung disease, including pneumonia, and chronic obstructive pulmonary disease, both of which are associated with bacterial infection. However, little is known about the effects of low dose arsenic exposure, or the contributions of organic arsenic to the innate immune response to bacterial infection. This study examined the effects on Pseudomonas aeruginosa (P. aeruginosa) induced cytokine secretion by human bronchial epithelial cells (HBEC) by inorganic sodium arsenite (iAsIII) and two major metabolites, monomethylarsonous acid (MMAIII) and dimethylarsenic acid (DMAV), at concentrations relevant to the U.S. population. Neither iAsIII nor DMAV altered P. aeruginosa induced cytokine secretion. By contrast, MMAIII increased P. aeruginosa induced secretion of IL-8, IL-6 and CXCL2. A combination of iAsIII, MMAIII and DMAV (10 pbb total) reduced IL-8 and CXCL1 secretion. These data demonstrate for the first time that exposure to MMAIII alone, and a combination of iAsIII, MMAIII and DMAV at levels relevant to the U.S. may have negative effects on the innate immune response of human bronchial epithelial cells to P. aeruginosa