Genomic and Microscopic Analysis of Ballast Water in the Great Lakes Region

Abstract

Invasive aquatic species can have damaging effects on fisheries and aquaculture through significant, and irreversible, effects on biodiversity. Human health may also be affected. To combat this threat the International Maritime Organization (IMO) Convention for Ballast Water and Sediments (BWMC) came into force in September 2017. U.S. Federal and IMO ballast water standards for discharged organisms stipulate discharge limits for different size classes of organisms. Several studies including recent trials aboard Great Lakes freighters have shown that many phytoplankton found in ballast water do not fall into the regulated 10–50 µM size class. Such issues illustrate the need for new methods of assessing microorganism populations that will supersede laborious microscopy requiring rare technical expertise. Recent progress has been made in the use of DNA (deoxyribose nucleic acid)-based methods as a means of identifying the appearance of invasive species in aquatic environments. A significant advance has been the development of high throughput sequencing (HTS), which has expanded DNA barcoding, relating to an individual organism, into second generation sequencing (metabarcoding), capable of mapping whole populations of organisms in an environmental sample. Several recent studies of HTS in ships’ ballast water, have shown that the technique has the capacity for detecting potentially harmful taxonomic groups and is capable of differentiating among water from different sources. The current study was undertaken to investigate the suitability (or otherwise) of HTS as a tool for ballast water management. Possible applications include improved risk assessment relating to invasive species. Feasibility for indicative testing for ballast water treatment efficacy was also addressed. However, pending analysis of treated samples, the current study was confined to a comparison of HTS and microscope counts in untreated samples. A correlation of visual and molecular taxonomic assignments of microorganisms found in the ballast water from different ports and during different seasons indicated that such a comparison was best conducted at Family level, although Principal Components Analysis showed that the two methods differed qualitatively among major taxonomic groups.