The impact of molecular biology on assessment of water quality: advantages and limitations of current techniques

The advent of molecular biology has had a dramatic impact on all aspects of biology, not least applied microbial ecology. Microbiological testing of water has traditionally depended largely on culture techniques. Growing understanding that only a small proportion of microbial species are culturable, and that many microorganisms may attain a viable but non-culturable state, has promoted the development of novel approaches to monitoring pathogens in the environment. This has been paralleled by an increased awareness of the surprising genetic diversity of natural microbial populations. By targeting gene sequences that are specific for particular microorganisms, for example genes that encode diagnostic enzymes, or species-specific domains of conserved genes such as 16S ribosomal RNA coding sequences (rrn genes), the problems of culture can be avoided. Technical developments, notably in the area of in vitro amplification of DNA using the polymerase chain reaction (PCR), now permit routine detection and identification of specific microorganisms, even when present in very low numbers. Although the techniques of molecular biology have provided some very powerful tools for environmental microbiology, it should not be forgotten that these have their own drawbacks and biases in sampling. For example, molecular techniques are dependent on efficient lysis and recovery of nucleic acids from both vegetative forms and spores of microbial species that may differ radically when growing in the laboratory compared with the natural environment. Furthermore, PCR amplification can introduce its own bias depending on the nature of the oligonucleotide primers utilised. However, despite these potential caveats, it seems likely that a molecular biological approach, particularly with its potential for automation, will provide the mainstay of diagnostic technology for the foreseeable future.

Evaluation of the In Situ Ichthyoplankton Imaging System (ISIIS): comparison with the traditional (bongo net) sampler

Plankton and larval fish sampling programs often are limited by a balance between sampling frequency (for precision) and costs. Advancements in sampling techniques hold the potential to add considerable efficiency and, therefore, add sampling frequency to improve precision. We compare a newly developed plankton imaging system, In Situ Ichthyoplankton Imaging System (ISIIS), with a bongo sampler, which is a traditional plankton sampling gear developed in the 1960s. Comparative sampling was conducted along 2 transects ~30–40 km long. Over 2 days, we completed 36 ISIIS tow-yo undulations and 11 bongo oblique tows, each from the surface to within 10 m of the seafloor. Overall, the 2 gears detected comparable numbers of larval fishes, representing similar taxonomic compositions, although larvae captured with the bongo were capable of being identified to lower taxonomic levels, especially larvae in the small (<5 mm), preflexion stages. Size distributions of the sampled larval fishes differed considerably between these 2 sampling methods, with the size range and mean size of larval fishes larger with ISIIS than with the bongo sampler. The high frequency and fine spatial scale of ISIIS allow it to add considerable sampling precision (i.e., more vertical sections) to plankton surveys. Improvements in the ISIIS technology (including greater depth of field and image resolution) should also increase taxonomic resolution and decrease processing time. When coupled with appropriate net sampling (for the purpose of collecting and verifying the identification of biological samples), the use of ISIIS could improve overall survey design and simultaneously provide detailed, process-oriented information for fisheries scientists and oceanographers.