Transmission of Chlamydia pneumoniae infection from blood monocytes to vascular cells in a novel transendothelial migration model

Chlamydia pneumoniae uses blood monocytes (PBMC) for systemic dissemination, persists in atherosclerotic lesions, and has been implicated in the pathogenesis of atherosclerosis. During transmigration in a newly developed transendothelial migration model (TEM) C. pneumoniae-infected PBMC spread their infection to endothelial cells. Transmigrated PBMC retained their infectivity and transmitted the pathogen to smooth muscle cells in the lower chamber of the TEM. Detection of chlamydial HSP60 mRNA proved pathogen viability and virulence. We conclude that PBMC can spread chlamydial infection to vascular wall cells and we suggest the TEM as a novel tool to analyze host-pathogen interactions in vascular chlamydial infections.     

Nocodazole inhibits macronuclear infection with Holospora obtusa in Paramecium caudatum

Summary. Holospora obtusa is a Gram-negative bacterium inhabiting the macronucleus of the ciliate Paramecium caudatum. Experimental infection with H. obtusa was carried out under nocodazole treatment. Nocodazole has been shown to cause disassembly of the cytoplasmic microtubules radiating from the cytopharynx and postoral fibers in P. caudatum. Treatment with this drug did not prevent the ingestion of both prey bacteria and H. obtusa, but it reduced the phagosome number and affected cyclosis. In situ hybridization revealed infectious forms of this endobiont very close to the macronucleus, but never inside it. These results indicate that disassembly of microtubules does not impair transportation of the infectious forms of H. obtusa in the cytoplasm, but that it completely blocks the invasion of the nucleus by the bacteria. Correspondence and reprints: Department of Cytology and Histology, Faculty of Biology and Soil Sciences, Saint Petersburg State University, Universitetskaya naberezhnaya 7/9, 199034 Saint Petersburg, Russia.     

Ferns and Melastomataceae as indicators of vascular plant composition in rain forests of Colombian Amazonia

In a case-study from Colombian Amazonia, species information from ferns and Melastomataceae was used to explain the compositional patterns of other vascular plant species in 40 widely distributed 0.1-ha plots. Canonical correspondence analysis was applied to regress vascular plant species composition in the forests against information from these two indicator groups (summarized as axes of principal coordinate analyses), together with that from soils, landscape, and the spatial sampling design. In total, 53,941 individuals of 2480 vascular plant species were recorded. Of these, 17,473 individuals and 132 species were from ferns and Melastomataceae. In 19 well-drained upland (tierra firme) plots 19,622 vascular plant individuals and 1716 species were found, with 3793 plants and 91 species from ferns and Melastomataceae. In both the set of all landscapes and the subset of tierra firme forests the principal PCoA axes of the two indicator groups were highly related to the main patterns of forest species composition. In principle, therefore, ferns and Melastomataceae can be used to detect and forecast changes in the forest composition of the study area. However, evidence was not obtained that ferns and Melastomataceae show more potential to predict the main patterns in species composition of forests than soil, landscape, and spatial variables. The partioning of the total variation in forest composition showed that the correlation of ferns and Melastomataceae with other forest plants was quite independent from that of soil, landscape, and space. Direct effects of ferns and Melastomataceae on other plants might be obtained from experimental studies of between-plant interactions, concentrating on the seedling or juvenile stages of trees and lianas, both above-ground as well as in the rooting environment.     

Susceptibility of juvenile Crassostrea gigas and resistance of Panope abrupta to Mikrocytos mackini

Samples from the field and laboratory exposure to Mikrocytos mackini (a tiny protistan parasite of unknown taxonomic affiliation) confirmed that juvenile Pacific oysters (Crassostrea gigas) are susceptible to infection and the resulting disease. In the laboratory bath exposure experiment, a prevalence of infection approaching 100% and mortalities were observed in the small oysters (about 18 mm in shell length). However, in the same laboratory exposure experiment, similar aged geoduck clams (Panope abrupta, about 8mm in shell length) were resistant to infection. The main route of infection in the oysters appeared to be via the digestive tract and possibly the gills where the parasite multiplied within host cells. Other tissues such as the adductor muscle and vesicular connective tissue were subsequently colonized. Although the infection resulted in the mortality of some oysters, others appeared to overcome the disease.     

Genetic variations in humans associated with differences in the course of hepatitis C

The outcome of hepatitis C virus (HCV) infection varies among individuals, but the genetic factors involved remain unknown. We conducted a population-based association study in which 238 Japanese individuals positive for anti-HCV antibody were genotyped for 269 single nucleotide polymorphisms (SNPs) in 103 candidate genes that might influence the course of infection. Altogether, 50 SNPs in 32 genes were listed. Genetic polymorphisms in IL4, IL8RB, IL10RA, PRL, ADA, NFKB1, GRAP2, CABIN1, IFNAR2, IFI27, IFI41, TNFRSF1A, ALDOB, AP1B1, SULT2B1, EGF, EGFR, TGFB1, LTBP2, and CD4 were associated with persistent viremia (P < 0.05), whereas those in IL1B, IL1RL1, IL2RB, IL12RB1, IL18R1, STAT5A, GRAP2, CABIN1, IFNAR1, Mx1, BMP8, FGL1, LTBP2, CD34, and CD80 were associated with different serum alanine aminotransferase levels in HCV carriers (P < 0.05). The sorted genes allow us to draw novel hypotheses for future studies of HCV infection to ultimately identify bona fide genes and their variations.     

Space-irrelevant scaling law for fish school sizes

Universal scaling in the power-law size distribution of pelagic fish schools is established. The power-law exponent of size distributions is extracted through the data collapse. The distribution depends on the school size only through the ratio of the size to the expected size of the schools an arbitrary individual engages in. This expected size is linear in the ratio of the spatial population density of fish to the breakup rate of school. By means of extensive numerical simulations, it is verified that the law is completely independent of the dimension of the space in which the fish move. Besides the scaling analysis on school size distributions, the integrity of schools over extended periods of time is discussed.     

A multiplex PCR assay to diagnose and quantify Nosema infections in honey bees (Apis mellifera)

Correct identification of the microsporidia, Nosema apis and Nosema ceranae, is key to the study and control of Nosema disease of honey bees (Apis mellifera). A rapid DNA extraction method combined with multiplex PCR to amplify the 16S rRNA gene with species-specific primers was compared with a previously published assay requiring spore-germination buffer and a DNA extraction kit. When the spore germination-extraction kit method was used, 10 or more bees were required to detect the pathogens, whereas the new extraction method made it possible to detect the pathogens in single bees. Approx. 4-8 times better detection of N. ceranae was found with the new method compared to the spore germination-extraction kit method. In addition, the time and cost required to process samples was lower with the proposed method compared to using a kit. Using the new DNA extraction method, a spore quantification procedure was developed using a triplex PCR involving co-amplifying the N. apis and N. ceranae 16S rRNA gene with the ribosomal protein gene, RpS5, from the honey bee. The accuracy of this semi-quantitative PCR was determined by comparing the relative band intensities to the number of spores per bee determined by microscopy for 23 samples, and a high correlation (R² = 0.95) was observed. This method of Nosema spore quantification revealed that spore numbers as low as 100 spores/bee could be detected by PCR. The new semi-quantitative triplex PCR assay is more sensitive, economical, rapid, simple, and reliable than previously published standard PCR-based methods for detection of Nosema and will be useful in laboratories where real-time PCR is not available.     

Activity of oil-formulated conidia of the fungal entomopathogens Nomuraea rileyi and Isaria tenuipes against lepidopterous larvae

The fungi Nomuraea rileyi and Isaria tenuipes (=Paecilomyces tenuipes) are ecologically obligate, widespread pathogens of lepidopterans. Bioassays were carried out to evaluate the activity of oil-suspended conidia of N. rileyi and I. tenuipes against larvae of Spodoptera frugiperda, Spodoptera exigua, Helicoverpa zea, and Heliothis virescens. The tests consisted of two bioassay sets. In the first set, conidia of N. rileyi and I. tenuipes were suspended in water + Tween 80, and in vegetable (canola, soybean) and mineral (proprietary mixture of alkanes and cyclic paraffins) oils, and tested against S. frugiperda. Both fungi were highly compatible with oils and caused mortalities near 100% in all oil treatments; the lowest LT₅₀ values were 4.7 days for N. rileyi in mineral oil and 6.0 days for I. tenuipes in soybean oil. The second set included additional fungal strains and oil formulations (mineral, canola, sunflower, olive and peanut oils) tested against larvae of S. exigua, S. frugiperda, H. zea and H. virescens. The highest activity was that of N. rileyi in mineral oil against Spodoptera spp., with LT₅₀ values of 2.5 days (strain ARSEF 135) and 3 days (strain ARSEF 762) respectively. For two different isolates of I. tenuipes the lowest LT₅₀ values (5.1-5.6 days respectively) were obtained with mineral oil formulations against Spodoptera spp. and H. zea respectively. Additionally, we tested both fungi against prepupae of all four lepidopteran species. Mortalities with I. tenuipes against S. exigua ranged from 90% to 100% (strains ARSEF 2488 and 4096); N. rileyi caused 95% mortality on S. frugiperda. The activity of formulations depended on host species and oil used; Spodoptera spp. was more susceptible to these fungi than Heliothis and Helicoverpa. The results indicate that a comprehensive evaluation of these entomopathogens in agriculture using oil application technologies is advisable, particularly, in organic and sustainable settings.     

Inhibition of Plasmodium sporozoites infection by targeting the host cell

There is a great need of new drugs against malaria because of the increasing spread of parasite resistance against the most commonly used drugs in the field. We found that monensin, a common veterinary antibiotic, has a strong inhibitory effect in Plasmodium berghei and Plasmodium yoelii sporozoites hepatocyte infection in vitro. Infection of host cells by another apicomplexan parasite with a similar mechanism of host cell invasion, Toxoplasma tachyzoites, was also inhibited. Treatment of mice with monensin abrogates liver infection with P. berghei sporozoites in vivo. We also found that at low concentrations monensin inhibits the infection of Plasmodium sporozoites by rendering host cells resistant to infection, rather than having a direct effect on sporozoites. Monensin effect is targeted to the initial stages of parasite invasion of the host cell with little or no effect on development, suggesting that this antibiotic affects an essential host cell component that is required for Plasmodium sporozoite invasion.     

Philopatry, nest choice, and aggregation temporal–spatial change in the digger wasp Cerceris arenaria (Hymenoptera: Crabronidae)

Studying temporal and spatial changes of aggregations of digger wasps through nesting seasons is interesting because of its link to social evolution via a semi-social pathway, but information about this topic is scarce. An aggregation of Cerceris arenaria Latreille was studied during 1997–1999 and for a short period in 2001. The colony grew in area and number of nests through the first three seasons, but the aggregation reduced in 2001, and this decrease was confirmed by a count of nests made in 2003 and 2005. The positions of the nest entrances remained almost constant over the years, at least in high-nest-density areas, because of the repeated use of the same burrows from which they emerged. This fact permits the population to exploit for many years a restricted area and could act as a nest-density-regulation method for fossorial species that rarely dig new nests, preventing an excessive nest density. The newly emerged females did not dig new nests, showed a high philopatry in the choice of their first nest, and preferred to look for other nests to occupy close to the ones recently abandoned (mainly for an undergone usurpation by a conspecific female). Philopatry, along with lack of new nest digging and position-dependent nest choice, is probably an important factor maintaining spatial stability of the aggregation through the years and could represent a first step in the evolution of a higher sociality in apoid burrowing Hymenoptera.