The inside scoop: Comparative genomics of two intranuclear bacteria, "Candidatus Berkiella cookevillensis" and "Candidatus Berkiella aquae".

"Candidatus Berkiella cookevillensis" (strain CC99) and "Candidatus Berkiella aquae" (strain HT99), belonging to the Coxiellaceae family, are gram-negative bacteria isolated from amoebae in biofilms present in human-constructed water systems. Both bacteria are obligately intracellular, requiring host cells for growth and replication. The intracellular bacteria-containing vacuoles of both bacteria closely associate with or enter the nuclei of their host cells. In this study, we analyzed the genome sequences of CC99 and HT99 to better understand their biology and intracellular lifestyles. The CC99 genome has a size of 2.9Mb (37.9% GC) and contains 2,651 protein-encoding genes (PEGs) while the HT99 genome has a size of 3.6Mb (39.4% GC) and contains 3,238 PEGs. Both bacteria encode high proportions of hypothetical proteins (CC99: 46.5%; HT99: 51.3%). The central metabolic pathways of both bacteria appear largely intact. Genes for enzymes involved in the glycolytic pathway, the non-oxidative branch of the phosphate pathway, the tricarboxylic acid pathway, and the respiratory chain were present. Both bacteria, however, are missing genes for the synthesis of several amino acids, suggesting reliance on their host for amino acids and intermediates. Genes for type I and type IV (dot/icm) secretion systems as well as type IV pili were identified in both bacteria. Moreover, both bacteria contain genes encoding large numbers of putative effector proteins, including several with eukaryotic-like domains such as, ankyrin repeats, tetratricopeptide repeats, and leucine-rich repeats, characteristic of other intracellular bacteria

Towards a Laser Beam Induced Current Test Structure for Nondestructive Determination of Junction Depth in HgCdTe Photodiodes

Correct placement of the p-n junction depth of diodes in HgCdTe infrared focal plane arrays is critical for ensuring the highest performance of diodes in the array. To date, most methods of determining the junction depth in HgCdTe have been destructive, based around wet chemical etching of the sample until the n-region has disappeared. In this paper we present some introductory work on the application of laser beam induced current, a non-destructive characterisation technique, to a specially designed junction depth test structure. The test structure has small geometric dimensions, which enables the peak magnitude of the LBIC signal to be sensitive to variations in the depth of the p-n junction

Infection and nuclear interaction in mammalian cells by ‘Candidatus Berkiella cookevillensis’, a novel bacterium isolated from amoebae

Abstract Background ‘Candidatus Berkiella cookevillensis’ and ‘Ca. Berkiella aquae’ have previously been described as intranuclear bacteria of amoebae. Both bacteria were isolated from amoebae and were described as appearing within the nuclei of Acanthamoeba polyphaga and ultimately lysing their host cells within 4 days. Both bacteria are Gammaproteobacteria in the order Legionellales with the greatest similarity to Coxiella burnetii. Neither bacterium grows axenically in artificial culture media. In this study, we further characterized ‘Ca. B. cookevillensis’ by demonstrating association with nuclei of human phagocytic and nonphagocytic cell lines. Results Transmission electron microscopy (TEM) and confocal microscopy were used to confirm nuclear co-localization of ‘Ca. B. cookevillensis’ in the amoeba host A. polyphaga with 100% of cells having bacteria co-localized with host nuclei by 48 h. TEM and confocal microscopy demonstrated that the bacterium was also observed to be closely associated with nuclei of human U937 and THP-1 differentiated macrophage cell lines and nonphagocytic HeLa human epithelial-like cells. Immunofluorescent staining revealed that the bacteria-containing vacuole invaginates the nuclear membranes and appears to cross from the cytoplasm into the nucleus as an intact vacuole. Conclusion Results of this study indicate that a novel coccoid bacterium isolated from amoebae can infect human cell lines by associating with the host cell nuclei, either by crossing the nuclear membranes or by deeply invaginating the nuclear membranes. When associated with the nuclei, the bacteria appear to be bound within a vacuole and replicate to high numbers by 48 h. We believe this is the first report of such a process involving bacteria and human cell lines

Rapid Response for Invasive Waterweeds at the Arctic Invasion Front: Assessment of Collateral Impacts from Herbicide Treatments

The remoteness of subarctic and arctic ecosystems no longer protects against invasive species introductions. Rather, the mix of urban hubs surrounded by undeveloped expanses creates a ratchet process whereby anthropogenic activity is sufficient to introduce and spread invaders, but for which the costs of monitoring and managing remote ecosystems is prohibitive. Elodea spp. is the first aquatic invasive plant to become established in Alaska and has potential for widespread deleterious ecological and economic impacts. A rapid eradication response with herbicides has been identified as a priority invasion control strategy. We conducted a multi-lake monitoring effort to assess collateral impacts from herbicide treatment for Elodea in high latitude systems. Variability in data was driven by seasonal dynamics and natural lake-to-lake differences typical of high latitude waterbodies, indicating lack of evidence for systematic impacts to water quality or plankton communities associated with herbicide treatment of Elodea. Impacts on native macrophytes were benign with the exception of some evidence for earlier onset of leaf senescence for lily pads(Nuphar spp.) in treated lakes. We observed a substantial increase in detected native flora richness after Elodea was eradicated from the most heavily infested lake, indicating potential for retention of native macrophyte communities if infestations are addressed quickly. While avoiding introductions through prevention may be the most desirable outcome, these applications indicated low risks of non-target impacts associated with herbicide treatment as a rapid response option for Elodea in high latitude systems