Two Homogametic Genotypes - One Crayfish: On the Consequences of Intersexuality.

In the Australian redclaw crayfish, Cherax quadricarinatus (WZ/ZZ system), intersexuals, although exhibiting both male and female gonopores, are functional males bearing a female genotype (WZ males). Therefore, the occurrence of the unusual homogametic WW females in nature is plausible. We developed W/Z genomic sex markers and used them to investigate the genotypic structure of experimental and native C. quadricarinatus populations in Australia. We discovered, for the first time, the natural occurrence of WW females in crustacean populations. By modeling population dynamics, we found that intersexuals contribute to the growth rate of crayfish populations in the short term. Given the vastly fragmented C. quadricarinatus habitat, which is characterized by drought-flood cycles, we speculate that intersexuals contribute to the fitness of this species since they lead to occasional increment in the population growth rate which potentially supports crayfish population restoration and establishment under extinction threats or colonization events.

Production of WW males lacking the masculine Z chromosome and mining the Macrobrachium rosenbergii genome for sex-chromosomes.

The cultivation of monosex populations is common in animal husbandry. However, preselecting the desired gender remains a major biotechnological and ethical challenge. To achieve an efficient biotechnology for all-female aquaculture in the economically important prawn (Macrobrachium rosenbergii), we achieved - for the first time - WW males using androgenic gland cells transplantation which caused full sex-reversal of WW females to functional males. Crossing the WW males with WW females yielded all-female progeny lacking the Z chromosome. We now have the ability to manipulate - by non-genomic means - all possible genotype combinations (ZZ, WZ and WW) to retain either male or female phenotypes and hence to produce monosex populations of either gender. This calls for a study of the genomic basis underlying this striking sexual plasticity, questioning the content of the W and Z chromosomes. Here, we report on the sequencing of a high-quality genome exhibiting distinguishable paternal and maternal sequences. This assembly covers ~ 87.5% of the genome and yielded a remarkable N50 value of ~ 20 × 106 bp. Genomic sex markers were used to initiate the identification and validation of parts of the W and Z chromosomes for the first time in arthropods.

Effect of treated domestic wastewater on soil physicochemical and microbiological properties.

A main concern with reuse of treated domestic wastewater (DWW) in irrigation is its possible effect on the soil. Few studies have focused on DWW treated in on-site settings, which generally use low-tech systems that can be constructed and serviced locally. One such system is the recirculating vertical flow constructed wetland (RVFCW). The aim of this study was to assess short- to midterm effects of irrigation with DWW treated in the RVFCW. Four groups of plastic barrels, filled with a sandy loam soil, were irrigated for 36 mo with fresh water (FW), FW with added fertilizer, raw DWW, or DWW treated in the RVFCW followed by ultraviolet disinfection. Principal component analysis revealed that the soil irrigated with treated DWW had physicochemical properties similar to those irrigated with FW amended with fertilizer. Levels of surfactants in soil irrigated with treated DWW were identical to those expected from standard irrigation practices, abating concerns for possible changes in soil hydraulic properties. was not detected in the soil irrigated with treated DWW, demonstrating the importance of disinfection of treated effluents before reuse in irrigation. Furthermore, irrigation with treated DWW did not alter the bacterial community structure according to terminal restriction fragment analysis. This 3-yr study suggests that the practice of irrigation with RVFCW effluents is safe. Continuation of the experiment is required to determine whether longer-term irrigation might show a different pattern.

Soil microbial abundance and diversity along a low precipitation gradient.

The exploration of spatial patterns of abundance and diversity patterns along precipitation gradients has focused for centuries on plants and animals; microbial profiles along such gradients are largely unknown. We studied the effects of soil pH, nutrient concentration, salinity, and water content on bacterial abundance and diversity in soils collected from Mediterranean, semi-arid, and arid sites receiving approximately 400, 300, and 100 mm annual precipitation, respectively. Bacterial diversity was evaluated by terminal restriction fragment length polymorphism and clone library analyses and the patterns obtained varied with the climatic regions. Over 75% of the sequenced clones were unique to their environment, while ∼2% were shared by all sites, yet, the Mediterranean and semi-arid sites had more common clones (∼9%) than either had with the arid site (4.7% and 6%, respectively). The microbial abundance, estimated by phospholipid fatty acids and real-time quantitative PCR assays, was significantly lower in the arid region. Our results indicate that although soil bacterial abundance decreases with precipitation, bacterial diversity is independent of precipitation gradient. Furthermore, community composition was found to be unique to each ecosystem.

Mathematical model for analysis of recirculating vertical flow constructed wetlands.

The recirculating vertical flow constructed wetland (RVFCW) was developed for the treatment of domestic wastewater (DWW). In this system, DWW is applied to a vertical flow bed through which it trickles into a reservoir located beneath the bed. It is then recirculated back to the root zone of the bed. In this study, a compartmental model was developed to simulate the RVFCW. The model, which addresses transport and removal kinetics of total suspended solids, 5-day biological oxygen demand and nitrogen, was fitted to kinetical results obtained from pilot field setups and a local sensitivity analysis was performed on the model parameters and operational conditions. This analysis showed that after 5h of treatment water quality is affected more by stochastic events than by the model parameter values, emphasizing the stability of the RVFCW system to large variations in operational conditions. Effluent quality after 1h of treatment, when the sensitivity analysis showed the parameter impacts to be largest, was compared to model predictions. The removal rate was found to be dependent on the recirculation rate. The predictions correlated well with experimental observations, leading to the conclusion that the proposed model is a satisfactory tool for studying RVFCWs.

Evaluating amplified rDNA restriction analysis assay for identification of bacterial communities.

Amplified ribosomal DNA restriction analysis (ARDRA) and restriction fragment length polymorphism were originally used for strain typing and for screening clone libraries to identify phylogenetic clusters within a microbial community. Here we used ARDRA as a model to examine the capacity of restriction-based techniques for clone identification, and the possibility of deriving phylogenetic information from ARDRA-based dendrograms. ARDRA was performed in silico on 48,759 sequences from the Ribosomal Database Project, and it was found that the fragmentation profiles were not necessarily unique for each sequence in the database, resulting in different species sharing fragmentation profiles. Although ARDRA-based clusters separated clones into different genera, these phylogenetic clusters did not overlap with trees constructed according to sequence alignment,calling into question the intra-genus ARDRA based phylogeny. It is thus suggested that the prediction power of ARDRA clusters in identifying clone phylogeny be regarded with caution.