Phycoflora of the tropical high-mountain lake El Sol,Central Mexico,and some biogeographical relationships

Algae collected from an oligotrophic crater lake were identified andcompared with similar regional information. A total of 50 genera and105 infrageneric taxa were identified, mostly Chlorophyta (66%),Cyanophyta (20%) and Bacillariophyta (7 genera: one central and sixpennates). Relatively few Chlorococcales were found in thisoligotrophic lake. The families most frequently found wereOedogoniaceae (18 species), Desmidiaceae (17), Zygnemataceae (13),Oscillatoriaceae (11), and Chaetophoraceae (9). Two genera ofCyanophyta, seven of Chlorophyta and 59 species are first records forMexico.     

Molecular profiling of diatom assemblages in tropical lake sediments using taxon-specific PCR and Denaturing High-Performance Liquid Chromatography (PCR-DHPLC)

Here we present a protocol to genetically detect diatoms in sediments of the Kenyan tropical Lake Naivasha, based on taxon-specific PCR amplification of short fragments (approximately 100 bp) of the small subunit ribosomal (SSU) gene and subsequent separation of species-specific PCR products by PCR-based denaturing high-performance liquid chromatography (DHPLC). An evaluation of amplicons differing in primer specificity to diatoms and length of the fragments amplified demonstrated that the number of different diatom sequence types detected after cloning of the PCR products critically depended on the specificity of the primers to diatoms and the length of the amplified fragments whereby shorter fragments yielded more species of diatoms. The DHPLC was able to discriminate between very short amplicons based on the sequence difference, even if the fragments were of identical length and if the amplicons differed only in a small number of nucleotides. Generally, the method identified the dominant sequence types from mixed amplifications. A comparison with microscopic analysis of the sediment samples revealed that the sequence types identified in the molecular assessment corresponded well with the most dominant species. In summary, the PCR-based DHPLC protocol offers a fast, reliable and cost-efficient possibility to study DNA from sediments and other environmental samples with unknown organismic content, even for very short DNA fragments.     

DNA-based analysis of planktonic methanotrophs in a stratified lake

1. The assemblage of planktonic methanotrophs in a stratified freshwater lake was investigated. Vertical patterns were analysed by denaturing gradient gel electrophoresis, using the primer pair specific for 16S rRNA genes of type I methanotrophs.
2. The resulting banding patterns could be divided into three distinct groups, and sequenced bands were all related to the Methylobacter species. No amplicon was obtained with the primer pair specific for type II methanotrophs.
3. Cloning analysis of the pmoA gene was performed using samples from three water depths (epilimnion, metalimnion and hypolimnion). The compositions of the clone libraries from the three depths were distinct from each other but all three libraries were dominated by clones related to Methylobacter species.     

Inhibition of quorum sensing in Chromobacterium violaceum by pigments extracted from Auricularia auricular

Aims: This study aimed to search for a novel quorum‐sensing inhibitor from some fungi and analyse its inhibitory activity. Methods and Results: Chromobacterium violaceum CV026, a double mini‐Tn5 mutant, was used as an indicator to monitor quorum‐sensing inhibition. Auricularia auricular pigments from fruiting bodies were extracted using hydrochloric acid as an infusion, dissolved in alkaline dimethylsulfoxide (DMSO), sterilized by filtration through a 0·22‐μm membrane filter and added to C. violaceum CV026 cultures. Inhibitory activity was measured by quantifying violacein production using a microplate reader. The results have revealed that the alkaline DMSO‐soluble pigments significantly reduced violacein production in a concentration‐dependent manner, a quorum‐sensing‐regulated behaviour in C. violaceum. Conclusions: Auricularia auricular pigments can inhibit bacterial quorum sensing. Significance and Impact of the Study: The results suggest the bioactive constituents from edible and medicinal fungi could interfere with bacterial quorum‐sensing system, regulate its associate functions and prevent bacterial pathogenesis. Further studies were in process in our laboratory to isolate specific compounds from A. auricular pigments, evaluate them as quorum‐sensing inhibitors and analyse the exact mechanism of action.     

Phytoplankton dynamics in a deep, tropical, hyposaline lake

The annual variation of the phytoplankton assemblage of deep (64.6 m), hyposaline (8.5 g l^–1) Lake Alchichica, central Mexico (19 ° N, 97° W), was analyzed in relation to thermal regime, and nutrients concentrations. Lake Alchichica is warm monomictic with a 3-month circulation period during the dry, cold season. During the stratified period in the warm, wet season, the hypolimnion became anoxic. N–NH₃ ranged between non detectable (n.d.) and 0.98 mg l^–1, N–NO₂ between n.d. and 0.007 mg l^–1, N–NO₃ from 0.1 to 1.0 mg l^–1 and P–PO₄ from n.d. to 0.54 mg l^–1. Highest nutrient concentrations were found in the circulation period. Chlorophyll a varied from <1 to 19.8 g l^–1 but most values were <5 g l^–1. The euphotic zone (>1% PAR) usually comprised the top 15–20 m. Nineteen algae species were identified, most of them are typical inhabitants of salt lakes. Diatoms showed the highest species number (10) but the small chlorophyte Monoraphidium minutum, the single-cell cyanobacteria, Synechocystis aquatilis, and the colonial chlorophyte, Oocystis parva, were the numerical dominant species over the annual cycle. Chlorophytes, small cyanobacteria and diatoms dominated in the circulation period producing a bloom comparable to the spring bloom in temperate lakes. At the end of the circulation and at the beginning of stratification periods, the presence of a bloom of the nitrogen-fixing cyanobacteria, N. spumigena, indicated nitrogen-deficit conditions. The well-stratified season was characterized by low epilimnetic nutrients levels and the dominance of small single-cell cyanobacteria and colonial chlorophytes. Phytoplankton dynamics in tropical Lake Alchichica is similar to the pattern observed in some deep, hyposaline, North American temperate lakes.     

A study on biological activity measurements and heterotrophic bacteria in a small freshwater lake

A 6-m-deep lake has been sampled to measure the temporal and depth-wise distribution of heterotrophic bacteria and biological activity in the water. Surface, mid-depth and bottom waters were analysed at monthly intervals for a period of one year. The coefficient of heterotrophic activity, alkaline phosphatase activity and biological oxygen demand are used as an index of biological activity. The bacterial community was at maximum during spring, coinciding with high values of biological activity. Highest biological activity was observed in the bottom waters. Dissolved organic carbon showed a significant positive correlation with most of the biological activity parameters. This suggests that biological activity, as measured by the coefficient of heterotrophic activity, was more closely related to the concentration of substrates than to population density of heterotrophic bacteria.     

Characterization of Streptococcus pneumoniae isolated from children with otitis media

Streptococcus pneumoniae is the main causative agent of acute otitis media in children. Serotype-based vaccines have provided some protection against otitis media, but not as much as anticipated, demonstrating the need for alternative vaccine options. Pneumococcal otitis media isolates were obtained from children 5 years old or younger from hospitals around Mississippi in the prevaccine era (1999-2000). These isolates were compared by capsular typing, pneumococcal surface protein A (PspA) family typing, antibiotic susceptibility, and DNA fingerprinting. Our study shows that there is great genetic variability among pneumococcal clinical isolates of otitis media, except with regard to PspA. Therefore, efforts focused on the development of a PspA-based pneumococcal vaccine would be well placed.     

Biosynthesis of a trypanocide by Chromobacterium violaceum

Radio-isotope studies indicated not only that l-tryptophan can serve as carbon source for synthesis of the trypanocide, violacein by Chromobacterium violaceum (BB-78 strain) but also that isatin and indole 3-acetic acid are both important metabolic intermediates. Using 3-indolyl [2-¹⁴C] and [1-¹⁴C] acetic acid, it was found that the carboxylic carbon was not eliminated and that indole-3-acetic acid was incorporated intact into the pigment structure. N-Ethyl(5-hydroxy-indol-3-yl)-2-indolylethylamide is also an important metabolic intermediate in the violacein biosynthesis. This is the first report of a metabolic scheme for violacein synthesis which includes an intermediate other than l-tryptophan.     

Seasonal dynamics,typhoons and the regulation of lake metabolism in a subtropical humic lake

1. We used high‐frequency in situ dissolved oxygen measurements to investigate the seasonal variability and factors regulating metabolism in a subtropical alpine lake in Taiwan between May 2004 and October 2005, specifically exploring how the typhoon season (from June or July to October) affects lake metabolism. 2. Gross primary production (GPP) and ecosystem respiration (R) both peaked in early summer and mid‐autumn but dropped during the typhoon season and winter. Yuan‐Yang Lake is a net heterotrophic ecosystem (annual mean net ecosystem production ?39.6 μmole O₂ m^?3). 3. Compared to the summer peaks, seasonal averages of GPP and R decreased by approximately 50% and 25%, respectively, during the typhoon season. Ecosystem respiration was more resistant to external disturbances than GPP and showed strong daily variation during typhoon seasons. 4. Changes in the quality and quantity of dissolved organic carbon controlled the temporal dynamics and metabolic regulation. External disturbances (typhoons) caused increased allochthony, increasing DOC and water colour and influencing lake metabolism. 5. Seasonal winter mixing and typhoon‐induced water mixing in summer and autumn play a key role in determining the extent to which the lake is a seasonal carbon sink or source to the atmosphere.     

The molecular diversity of the methanogenic community in a hypereutrophic freshwater lake determined by PCR-RFLP

AIMS: To combine database-held sequence information with a programme of experimental molecular ecology to define the methanogenic community of a hypereutrophic lake by a PCR-restriction fragment length polymorphism (RFLP) analysis. METHODS AND RESULTS: Methanogen diversity in a hypereutrophic freshwater lake was analysed using 16S rDNA PCR-RFLP. Database-held 16S rRNA gene sequences for 76 diverse methanogens were analysed for specific restriction sites that permitted unequivocal differentiation of methanogens. Restriction digestion and agarose gel electrophoresis of the 16S rDNA from selected methanogen pure cultures generated observed restriction profiles that corroborated the expected patterns. This method was then tested by analysing methanogen diversity in samples obtained over 1 year from sediment and water samples taken from the same sampling site. CONCLUSIONS: Restriction analysis of the 16S rRNA gene sequences from 157 methanogen clones generated from lakewater and sediment samples showed that over 50% were similar to Methanoculleus spp. Furthermore, a total of 16 RFLP types (1-16) were identified, eight of which contained no cultured representative archaeal 16S rRNA gene sequences. SIGNIFICANCE AND IMPACT OF THE STUDY: This RFLP strategy provides a robust and reliable means to rapidly identify methanogens in the environment.     

Biosynthesis of novel polyhydroxyalkanoate containing 3-hydroxy-4-methylvalerate by Chromobacterium sp. USM2

Aims: Polyhydroxyalkanoate (PHA) with enhanced physicochemical properties will be ideal for a wide range of practical applications. The incorporation of 3‐hydroxy‐4‐methylvalerate (3H4MV) into the polymer backbone is known to improve the overall properties of the resulting polymer. However, the most suitable micro‐organism and PHA synthase that can synthesize this monomer efficiently still remain unknown at present. Therefore, we evaluated the abilities of a locally isolated Chromobacterium sp. USM2 to produce PHA containing 3H4MV. Methods and Results: The ability of Chromobacterium sp. USM2 to synthesize poly(3‐hydroxybutyrate‐co‐3‐hydroxy‐4‐methylvalerate) [P(3HB‐co‐3H4MV)] was evaluated under different culture conditions. It was found that Chromobacterium sp. USM2 can synthesize P(3HB‐co‐3H4MV) when glucose and isocaproic acid were fed as carbon source. However, the highest molar fraction of 3H4MV, 22 mol% was detected in Chromobacterium sp. USM2 when isocaproic acid was provided as the sole carbon source. In addition, aeration was identified as a crucial factor in initiating the accumulation of high 3H4MV molar fractions. Conclusions: Chromobacterium sp. USM2 was able to synthesize broad comonomer compositional distribution of P(3HB‐co‐3H4MV). Significance and Impact of the Study: Compared with Cupriavidus necator and Burkholderia sp., Chromobacterium sp. USM2 was found to have better ability to bioconvert isocaproic acid to form 3H4MV unit.     

Vertical diffusion and nutrient transport in a tropical lake (Lake McIlwaine,Rhodesia)

Measurements of the vertical temperature in tropical Lake McIlwaine were used to calculate the time-averaged ( 6 months) vertical diffusivity coefficient (K_z) in the metalimnion and hypolimnion. The mean value of K_z (0.21 cm² s^–1) was correlated with the lake surface area. The mass transport rates of PO₄-P and NH₄-N, upward from the hypolimnion to the metalimnion, were calculated using K_z and measured values of the nutrient concentration gradients. During a period of 4.5 months when the water was stably stratified, PO₄-P was transported upward at a mean rate of 42 kg day^–1 and NH₄-N at a mean rate of 162 kg day^–1 over the entire lake.     

Quorum sensing is a key regulator for the antifungal and biocontrol activity of chitinase‐producing Chromobacterium sp. C61

Chromobacterium sp. strain C61 has strong biocontrol activity; however, the genetic and biochemical determinants of its plant disease suppression activity are not well understood. Here, we report the identification and characterization of two new determinants of its biocontrol activity. Transposon mutagenesis was used to identify mutants that were deficient in fungal suppression. One of these mutants had an insertion in a homologue of depD, a structural gene in the dep operon, that encodes a protein involved in non‐ribosomal peptide synthesis. In the second mutant, the insertion was in a homologue of the luxI gene, which encodes a homoserine lactone synthase. The luxI^– and depD^– mutants had no antifungal activity in vitro and a dramatically reduced capacity to suppress various plant diseases in planta. Antifungal production and biocontrol were restored by complementation of the luxI^– mutant. Other phenotypes associated with effective biological control, including motility and lytic enzyme secretion, were also affected by the luxI mutation. Biochemical analysis of ethyl acetate extracts of culture filtrates of the mutant and wild‐type strains showed that a key antifungal compound, chromobactomycin, was produced by wild‐type C61 and the complemented luxI^– mutant, but not by the luxI^– or depD^– mutant. These data suggest that multiple biocontrol‐related phenotypes are regulated by homoserine lactones in C61. Thus, quorum sensing plays an essential role in the biological control potential of diverse bacterial lineages.     

Dynamic of phytoplankton size-class and photosynthetic activity in a tropical hypereutrophic lake: the Yaounde municipal lake (Cameroon)

This study carried out within the framework of a multi-disciplinary project, aimed at highlighting the fundamental processes governing the functioning of the hypertrophic Yaoundé Municipal Lake. It was based on the hypothesis that, even within a small range of lake trophic status, important variations can occur in the species composition, biomass and photosynthetic activity of the phytoplankton size-fractions. For this purpose, samples were taken at weekly intervals from November 1996 to December 1997 at a fixed set of depths in the water column. Spatial-temporal fluctuations of some physical–chemical variables, associated with biological variables such as phytoplankton size-class species composition, phytoplankton size-class biomass, phytoplankton primary production and chlorophyll-a were analysed. The water transparency was low and rarely exceeded 100 cm. Conductivity values relatively higher increased generally from the top to the lake bottom. The oxygen deficiency, and sometimes anoxia, recorded from 2.5 m depth resulted in high quantities of ammonium nitrogen. Total phosphorus and total Kjeldahl nitrogen concentrations were characteristics of hypertrophic lakes. The fertility of this biotope favoured the development of a high phytoplanktonic community with remarkable physiological adaptations to the variations of the nutritive potentials of the lake, characterized by the size-structure of these organisms. Cells of small size (<12 μm) contributed up to 11.42% of the total phytoplanktonic biomass. Species with average size (12–45 μm), dominated by Chlamydomonas spp., represented a more significant contribution reaching up to 69.85%, whereas the cells of big size (>45 μm), mainly Euglenophyta, maintained the relatively most important biomass, accounting for up to 89.85% of the total phytoplanktonic biomass. Chlorophyll-a concentrations are among the highest reported for both fresh water and sea water, being a consequence of high proportions of Chlorophyta and Euglenophyta. This led to intense phytoplanktonic photosynthetic activity which continued throughout the year, even though it was confined to the upmost first meter of the water column. Analyses pointed out the allogenic nature of the functioning of this urban lake ecosystem, due to a poor waste management on the surrounding landscape. Handling editor: J. Padisak     

Boeckella diamantina n.sp. (Calanoida,Centropagidae), from a high Andean lake in Mendoza,Argentina

A new species of Boeckella from limnetic samples of Laguna del Diamante, a high lake in the Andes (34°10 S) is described and illustrated. The species is defined by the characters of the male fifth leg: the right two segmented endopod bears four peculiar, short, claw-like spines, the left endopod is a simple finger-like projection. This species is related to B. gibbosa, also a species from the Andes and B. vallentini from Malvinas (Falkland Islands) and other subantarctic islands. It is distinguished from them by diagnostic features of the fifth legs of the male and abdominal structure and fifth legs of the female. Some current views on the features used in the taxonomy of the genus Boeckella are discussed.     

Swamp-riverine tropical fish population: a comparative study of two spatially isolated freshwater ecosystems in Peninsular Malaysia

Twenty-four fish species were collected during a study from October 1996 of the swamp-riverine fish populations of two spatially isolated Malaysian freshwater swamp ecosystems. Twenty one species were collected from the Beriah Kanan River, which is located in northern Peninsular Malaysia, compared to only 10 species obtained from the Ulu Sedili River in the south. At both sites, the species Rasbora einthovenii and Hemirhamphodon pogognathu appeared to be the most dominant species, whereas Rasbora pauciperforata and Nemacheilus selangoricus were very rarely caught and Beriah Kanan and Ulu Sedili River, respectively. An analysis of the physico-chemical data showed that although pH, conductivity and ammonium levels were fairly similar at both sites, orthophosphate, hardness and total suspended solids were relatively higher for Ulu Sedili River.