DNA Probe-Mediated Detection of Resistant Bacteria from Soils Highly Polluted by Heavy Metals

Alcaligenes eutrophus CH34 DNA fragments encoding resistance to Cd²⁺, Co²⁺, Zn²⁺ (czc), or Hg²⁺ (merA) were cloned and used as probes in colony hybridization procedures with bacteria isolated from polluted environments such as a zinc factory area (desertified because of the toxic effects of zinc contamination) and from sediments from factories of nonferrous metallurgy in Belgium and mine areas in Zaire. From the different soil samples, strains could be isolated and hybridized with the czc probe (resistance to Cd²⁺, Co²⁺, and Zn²⁺ from plasmid pMOL30). Percentages of CFU isolated on nonselective plates which hybridized with the czc and the mercury resistance probes were, respectively, 25 and 0% for the zinc desert, 15 to 20 and 10 to 20% for the two Belgian factories, and 40 and 40% for the Likasi mine area. Most of these strains also carried two large plasmids of about the same size as those of A. eutrophus CH34 and shared many phenotypic traits with this strain. These findings indicated a certain correlation between the heavy-metal content in contaminated soils and the presence of heavy-metal-resistant megaplasmid-bearing A. eutrophus strains.     

Inhibition of dopamine receptors in the stomach: An explanation of the gastrokinetic properties of domperidone

We investigated the effects of dopamine and secrenin on the motility of the isolated guinea pig stomach. Changes in intragastric volume were continously recorded as a measure of gastric motility. When 0.25 μg/ml dopamine was given intra-arterially (i.a.) via the coeliac axis, the stomach relaxed, its spontaneous activity decreased, and its content augmented. This effect of dopamine could be selectively inhibited by the dopamine antagonists haloperidol (0.16 μg/ml) and domperidone (0.16 μg/ml); the latter is effective in the treatment of gastro-intestinal dysfunction. These observations demonstrate the presence of dopaminergic receptors in the guinea pig stomach. Similar conclusions can be reached for the rat stomach from ³H-domperidone displacement studies. Secretin (6.7 mU/ml i.a.) inhibited the vagally induced emptying of the stomach. This effect is reversed by domperidone (0.04 μg/ml). These results strongly suggest that dopamine or/and secretin are involved in the local feedback control of gastric motility; the action of secretin may be direct or indirect by release of dopamine. The local interference of domperidone with endogenous secretin and dopamine in the stomach may explain its gastrokinetic properties.     

Dynamics of an Oligotrophic Bacterial Aquifer Community during Contact with a Groundwater Plume Contaminated with Benzene, Toluene, Ethylbenzene, and Xylenes: an In Situ Mesocosm Study

An in situ mesocosm system was designed to monitor the in situ dynamics of the microbial community in polluted aquifers. The mesocosm system consists of a permeable membrane pocket filled with aquifer material and placed within a polypropylene holder, which is inserted below groundwater level in a monitoring well. After a specific time period, the microcosm is recovered from the well and its bacterial community is analyzed. Using this system, we examined the effect of benzene, toluene, ethylbenzene, and xylene (BTEX) contamination on the response of an aquifer bacterial community by denaturing gradient gel electrophoresis analysis of PCR-amplified 16S rRNA genes and PCR detection of BTEX degradation genes. Mesocosms were filled with nonsterile or sterile aquifer material derived from an uncontaminated area and positioned in a well located in either the uncontaminated area or a nearby contaminated area. In the contaminated area, the bacterial community in the microcosms rapidly evolved into a stable community identical to that in the adjacent aquifer but different from that in the uncontaminated area. At the contaminated location, bacteria with tmoA- and xylM/xylE1-like BTEX catabolic genotypes colonized the aquifer, while at the uncontaminated location only tmoA-like genotypes were detected. The communities in the mesocosms and in the aquifer adjacent to the wells in the contaminated area consisted mainly of Proteobacteria. At the uncontaminated location, Actinobacteria and Proteobacteria were found. Our results indicate that communities with long-term stability in their structures follow the contamination plume and rapidly colonize downstream areas upon contamination.     

Isolation of Adherent Polycyclic Aromatic Hydrocarbon (PAH)-Degrading Bacteria Using PAH-Sorbing Carriers

Two different procedures were compared to isolate polycyclic aromatic hydrocarbon (PAH)-utilizing bacteria from PAH-contaminated soil and sludge samples, i.e., (i) shaken enrichment cultures in liquid mineral medium in which PAHs were supplied as crystals and (ii) a new method in which PAH degraders were enriched on and recovered from hydrophobic membranes containing sorbed PAHs. Both techniques were successful, but selected from the same source different bacterial strains able to grow on PAHs as the sole source of carbon and energy. The liquid enrichment mainly selected for Sphingomonas spp., whereas the membrane method exclusively led to the selection of Mycobacterium spp. Furthermore, in separate membrane enrichment set-ups with different membrane types, three repetitive extragenic palindromic PCR-related Mycobacterium strains were recovered. The new Mycobacterium isolates were strongly hydrophobic and displayed the capacity to adhere strongly to different surfaces. One strain, Mycobacterium sp. LB501T, displayed an unusual combination of high adhesion efficiency and an extremely high negative charge. This strain may represent a new bacterial species as suggested by 16S rRNA gene sequence analysis. These results indicate that the provision of hydrophobic sorbents containing sorbed PAHs in the enrichment procedure discriminated in favor of certain bacterial characteristics. The new isolation method is appropriate to select for adherent PAH-degrading bacteria, which might be useful to biodegrade sorbed PAHs in soils and sludge.     

Roost and hunting site fidelity of female and juvenile Daubenton's bat Myotis daubentonii (Kuhl, 1817) (Chiroptera: Vespertilionidae)

We investigated roosting and hunting site fidelity of Daubenton's bats Myotis daubentonii (Kuhl, 1817) in the Forêt de Soignes, an old-stand forest dominated by 150–200-year-old beeches, during the summers of 2003 and 2004. Roosting behaviour and hunting activity over ponds of adult females and juveniles were monitored using radio-telemetry. Eighteen roosts were located, all in natural cavities. The bats occupied a limited number of trees located in a specific and small roosting area. This roost aggregation was not linked to the distribution of hollow trees. Furthermore, whereas all 11 ponds in the study area were exploited by Daubenton's bats, monitored individuals were limited to two ponds to which they showed high fidelity. These two ponds were not the closest to the roosting area. Overall, these results show that at least for the time we conducted our experiment, female and juvenile Daubenton's bats are highly faithful to specific roosting and hunting grounds.     

Selection for human immunodeficiency virus type 1 envelope glycosylation variants with shorter V1-V2 loop sequences occurs during transmission of certain genetic subtypes and may impact viral RNA levels

Designing an effective human immunodeficiency virus type 1 (HIV-1) vaccine will rely on understanding which variants, from among the myriad of circulating HIV-1 strains, are most commonly transmitted and determining whether such variants have an Achilles heel. Here we show that heterosexually acquired subtype A HIV-1 envelopes have signature sequences that include shorter V1-V2 loop sequences and fewer predicted N-linked glycosylation sites relative to the overall population of circulating variants. In contrast, recently transmitted subtype B variants did not, and this was true for cases where the major risk factor was homosexual contact, as well as for cases where it was heterosexual contact. This suggests that selection during HIV-1 transmission may vary depending on the infecting subtype. There was evidence from 23 subtype A-infected women for whom there was longitudinal data that those who were infected with viruses with fewer potential N-linked glycosylation sites in V1-V2 had lower viral set point levels. Thus, our study also suggests that the extent of glycosylation in the infecting virus could impact disease progression.     

Infection with multiple human immunodeficiency virus type 1 variants is associated with faster disease progression

Human immunodeficiency virus type 1 (HIV-1)-infected individuals develop a genetically diverse virus population over time, but often only a limited number of viral variants are transmitted from a chronic carrier to a newly infected person. Interestingly, many women but few men are infected by multiple HIV-1 variants from a single partner. To determine whether the complexity of the infecting virus population influences clinical outcome, we examined viral diversity in the HIV-1 envelope sequences present at primary infection in 156 women from Kenya for whom we had follow-up data on viral RNA levels and CD4 T-cell counts. Eighty-nine women had multiple viral genotypes, while 67 women had a single genotype at primary infection. Women who acquired multiple viral genotypes had a significantly higher viral load (median, 4.84 versus 4.64 log(10) copies/ml, P = 0.04) and a significantly lower CD4(+)-T-cell count (median, 416 versus 617 cells/mm(3), P = 0.01) 4 to 24 months after infection compared to women who were infected with a single viral genotype. These studies suggest that early HIV-1 genetic diversity is linked to faster disease progression.     

Individual-and population-based diversity in restriction-modification systems

Restriction-modification (RM) systems are cognate gene complexes that code for an endonuclease and a methylase. They are often thought to have developed in bacteria as protection against invading genetic material, e.g., phage DNA. The high diversity of RM systems, as observed in nature, is often ascribed to the coevolution of RM systems (which ‘invent’ novel types) and phages. However, the extent to which phages are insensitive to RM systems casts doubts on the effectiveness of RM systems as protection against infection and thereby on the reason for the diversity of RM systems. We present an eco-evolutionary model in order to study the evolution of the diversity of RM systems. The model predicts that in general diversity of RM systems is high. More importantly, the diversity of the RM systems is expressed either at the individual level or at the population level. In the first case all individuals carry RM systems of all sequence specificities, whereas in the second case they carry only one RM system or no RM systems at all. Nevertheless, in the second case the same number of sequence specificities are present in the population.