Bonamia exitiosa in European Flat Oyster (Ostrea edulis) on the Croatian Adriatic Coast from 2016 to 2020

The annual production of European flat oysters (Ostrea edulis) in Croatia is about 50 to 65 tons, and it has a long tradition. All Croatian oyster farms are subjected to the national surveillance program aiming to detect the presence of Bonamia ostreae and Marteilia refringens according to the Council Directive 2006/88/EC. Within the surveillance program, the first findings of the parasite Bonamia spp. occurred in 2016 in two production areas in the north and south of the Eastern Adriatic coast. The repeated findings of the parasite were noted up to 2020 but also on two additional sites in the north. The parasite was detected by cytological analysis of stained heart smears, histological examination, and PCR. PCR positive samples were sequenced for SSU rDNA gene, and BLAST analysis confirmed infection with Bonamia exitiosa. Attempts to prove the Pacific oyster as a putative vector of the parasite failed. The infection prevalence from 2016 until 2020 ranged from 3.3 to 20% in different sites. No mortalities were reported from the infected sites, and it seemed that infection of flat oysters with B. exitiosa did not affect their health. The study has not shown the source and way of infection spread, which imposes the need for more comprehensive molecular and epidemiological studies.

A non-lethal method for detection of Bonamia ostreae in flat oyster (Ostrea edulis) using environmental DNA

Surveillance and diagnosis of parasitic Bonamia ostreae infections in flat oysters (Ostrea edulis) are prerequisites for protection and management of wild populations. In addition, reliable and non-lethal detection methods are required for selection of healthy brood oysters in aquaculture productions. Here we present a non-lethal diagnostic technique based on environmental DNA (eDNA) from water samples and demonstrate applications in laboratory trials. Forty oysters originating from Limfjorden, Denmark were kept in 30 ppt sea water in individual tanks. Water was sampled 6 days later, after which all oysters were euthanized and examined for infection, applying PCR. Four oysters (10%) were found to be infected with B. ostreae in gill and mantle tissue. eDNA purified from the water surrounding these oysters contained parasite DNA. A subsequent sampling from the field encompassed 20 oysters and 15 water samples from 5 different locations. Only one oyster turned out positive and all water samples proved negative for B. ostreae eDNA. With this new method B. ostreae may be detected by only sampling water from the environment of isolated oysters or isolated oyster populations. This non-lethal diagnostic eDNA method could have potential for future surveys and oyster breeding programs aiming at producing disease-free oysters.

A Naïve Population of European Oyster Ostrea edulis with Reduced Susceptibility to the Pathogen Bonamia ostreae: Are S-Strategy Life Traits Providing Protection?

Synopsis European populations of the native flat oyster, Ostrea edulis, have been heavily depleted by two protozoan parasites, Marteila refringens and Bonamia ostreae, with mortalities of up to 90% reported in naïve populations. However, in studies carried out over a 10-year period, researching the parasite–host relationship of B. ostreae and O. edulis in several age cohorts within a naïve O. edulis population from Loch Ryan (LR), Scotland, 1364 specimens were challenged and only 64 (5%), across multiple testing protocols, screened positive for B. ostreae. This article presents a case for the development of S-strategy life traits in the LR population that coincide with enhanced immune function and survival. Oysters are considered typical r-strategists (small in size with fast development and high fecundity) while S-strategists, as outlined in Grime’s (1977) competitor–stress tolerant–ruderal (C–S–R) triangle theory, are characterized by slow growth and investment in the durability of individuals. This study hypothesizes that slower growth and reduced reproductive output in LR oysters has resulted in the investment of an enhanced immune function and reduced susceptibility to B. ostreae that is, r-strategists with S-strategy life traits equates to protection from significant pathogens. The findings presented here within provide a strong case study for local adaptation of energy allocation and provides empirical support for the C–S–R triangle theory in a marine organism.

Low internal transcribed spacer rDNA variation in New Zealand Bonamia ostreae: evidence for a recent arrival

Bonamia ostreae is a haplosporidian parasite of oysters that was first reported to occur in the Southern Hemisphere in 2015 in the New Zealand flat oyster Ostrea chilensis. Until that report, B. ostreae had been restricted to populations of O. edulis within the Northern Hemisphere. This large range extension raised questions regarding B. ostreae dispersal, including whether B. ostreae is a recent introduction and from where it originated. The whole 18S rRNA gene of New Zealand B. ostreae revealed 99.9-100% sequence homology to other published B. ostreae 18S rDNA sequences. Internal transcribed spacer (ITS) rDNA sequences (n = 29) were generated from New Zealand B. ostreae and compared to published B. ostreae sequences from 3 Northern Hemisphere sites: California, USA (n = 18), Maine, USA (n = 7), and the Netherlands (n = 6) to investigate intraspecific variation. Low ITS rDNA variation was observed from New Zealand B. ostreae isolates, and high levels of variation were observed from Northern Hemisphere B. ostreae sequences. We hypothesise that the low ITS rDNA diversity found in New Zealand B. ostreae is the result of a founder effect resulting from a single introduction from a limited number of propagules. The high level of ITS rDNA variation from the Northern Hemisphere prevented inferences of dispersal origins. New Zealand B. ostreae were genetically differentiated from all sites, and additional genetic data are required to better determine the origin of B. ostreae in New Zealand.