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Thierry M. Work

Publications and source records attributed to Thierry M. Work.

At least 127 records · Page 7Linked to original sources

Microbial disease and the coral holobiont

Tropical coral reefs harbour a reservoir of enormous biodiversity that is increasingly threatened by direct human activities and indirect global climate shifts. Emerging coral diseases are one serious threat implicated in extensive reef deterioration through disruption of the integrity of the coral holobiont – a complex symbiosis between the coral animal, endobiotic alga and an array of microorganisms. In this article, we review our current understanding of the role of microorganisms in coral health and disease, and highlight the pressing interdisciplinary research priorities required to elucidate the mechanisms of disease. We advocate an approach that applies knowledge gained from experiences in human and veterinary medicine, integrated into multidisciplinary studies that investigate the interactions between host, agent and environment of a given coral disease. These approaches include robust and precise disease diagnosis, standardised ecological methods and application of rapidly developing DNA, RNA and protein technologies, alongside established histological, microbial ecology and ecological expertise. Such approaches will allow a better understanding of the causes of coral mortality and coral reef declines and help assess potential management options to mitigate their effects in the longer term.

Trends in Microbiology

Identification of CD3+ T lymphocytes in the green turtle Chelonia mydas

To understand the role of the immune system with respect to disease in reptiles, there is the need to develop tools to assess the host's immune response. An important tool is the development of molecular markers to identify immune cells, and these are limited for reptiles. We developed a technique for the cryopreservation of peripheral blood mononuclear cells and showed that a commercially available anti-CD3 epsilon chain antibody detects a subpopulation of CD3 positive peripheral blood lymphocytes in the marine turtle Chelonia mydas . In the thymus and in skin inoculated with phytohemagglutinin, the same antibody showed the classical staining pattern observed in mammals and birds. For Western blot, the anti-CD3 antibodies identified a 17.6 kDa band in membrane proteins of peripheral blood mononuclear cell compatible in weight to previously described CD3 molecules. This is the first demostration of CD3+ cells in reptiles using specific antibodies.

Quintana Roo

In vitro biology of fibropapilloma-associated turtle herpesvirus and host cells in Hawaiian green turtles (Chelonia mydas)

Fibropapillomatosis (FP) of green turtles has a global distribution and causes debilitating tumours of the skin and internal organs in several species of marine turtles. FP is associated with a presently non-cultivable alphaherpesvirus Chelonid fibropapilloma-associated herpesvirus (CFPHV). Our aims were to employ quantitative PCR targeted to pol DNA of CFPHV to determine (i) if DNA sequesters by tumour size and/or cell type, (ii) whether subculturing of cells is a viable strategy for isolating CFPHV and (iii) whether CFPHV can be induced to a lytic growth cycle in vitro using chemical modulators of replication (CMRs), temperature variation or co-cultivation. Additional objectives included determining whether non-tumour and tumour cells behave differently in vitro and confirming the phenotype of cultured cells using cell-type-specific antigens. CFPHV pol DNA was preferentially concentrated in dermal fibroblasts of skin tumours and the amount of viral DNA per cell was independent of tumour size. Copy number of CFPHV pol DNA per cell rapidly decreased with cell doubling of tumour-derived fibroblasts in culture. Attempts to induce viral replication in known CFPHV-DNA-positive cells using temperature or CMR failed. No significant differences were seen in in vitro morphology or growth characteristics of fibroblasts from tumour cells and paired normal skin, nor from CFPHV pol-DNA-positive intestinal tumour cells. Tumour cells were confirmed as fibroblasts or keratinocytes by positive staining with anti-vimentin and anti-pancytokeratin antibodies, respectively. CFPHV continues to be refractory to in vitro cultivation.

Hawai'i

Rise and fall over 26 years of a marine epizootic in Hawaiian green sea turtles

Estimates of chronic disease prevalence are needed to improve our understanding of marine disease epizootiology, which is poorly known for marine megafauna such as marine turtles. An emerging worldwide threat to green sea turtles ( Chelonia mydas ) is fibropapillomatosis (FP), which is a pandemic tumor-forming disease associated with herpesviruses. We report on a 26-yr FP epidemic in the Hawaiian Archipelago and show that apparent disease prevalence in the world's main endemic hot spot increased rapidly following a late 1980s outbreak, peaked during the mid-1990s, and then declined steadily ever since. While this disease is a major cause of sea turtle stranding in Hawaiian waters and can be fatal, we also show that long-term tumor regression can occur even for turtles with advanced FP. The endemic Hawaiian green turtle stock was severely depleted by overexploitation prior to protection under the US Endangered Species Act in 1978. This stock has increased significantly ever since, despite exposure to a major chronic disease epidemic that is currently declining.

Journal of Wildlife Diseases

Phase shift from a coral to a corallimorph-dominated reef associated with a shipwreck on Palmyra atoll

Coral reefs can undergo relatively rapid changes in the dominant biota, a phenomenon referred to as phase shift. Various reasons have been proposed to explain this phenomenon including increased human disturbance, pollution, or changes in coral reef biota that serve a major ecological function such as depletion of grazers. However, pinpointing the actual factors potentially responsible can be problematic. Here we show a phase shift from coral to the corallimorpharian Rhodactis howesii associated with a long line vessel that wrecked in 1991 on an isolated atoll (Palmyra) in the central Pacific Ocean. We documented high densities of R. howesii near the ship that progressively decreased with distance from the ship whereas R. howesii were rare to absent in other parts of the atoll. We also confirmed high densities of R. howesii around several buoys recently installed on the atoll in 2001. This is the first time that a phase shift on a coral leef has been unambiguously associated with man-made structures. This association was made, in part, because of the remoteness of Palmyra and its recent history of minimal human habitation or impact. Phase shifts can have long-term negative ramification for coral reefs, and eradication of organisms responsible for phase shifts in marine ecosystems can be difficult, particularly if such organisms cover a large area. The extensive R. howesii invasion and subsequent loss of coral reef habitat at Palmyra also highlights the importance of rapid removal of shipwrecks on corals reefs to mitigate the potential of reef overgrowth by invasives.

Palmyra Atoll

Overgrowth of fungi (endolithic hypermycosis) associated with multifocal to diffuse distinct amorphous dark discoloration of corals in the Indo-Pacific

Coral disease surveys in American Samoa and Hawai‘i revealed colonies with a distinct dark discoloration affecting 20–60% of the colony surface (Fig. 1 a). In some cases, tissue loss with algal infiltration was present within discolored areas. On microscopy, these lesions had marked overgrowth of the coral skeleton and tissues with septate branching structures that stained positive with Grocott’s Methenamine Silver (fungal hyphae) accompanied by necrosis and fragmentation of coral tissues (Fig. 1 b). We have observed this condition grossly and microscopically in Pavona varians , Psammocora nierstraszi , and Montipora sp. in American Samoa and in Pavona maldivensis and P. varians in Hawai‘i. This condition resembles Dark Spots Disease from the Caribbean (Solano et al. 1993 ) that also shows endolithic hypermycosis (Galloway et al. 2007 ), suggesting that the association between dark discoloration of corals and overgrowth of endolithic fungi may be common (Western Atlantic, Indo-Pacific). Based on gross and microscopic morphology, tissue atrophy may precede overgrowth of endolithic fungi, but this awaits confirmation through systematic studies that monitor the development of lesions over time (pathogenesis). Using standardized terminology (Work and Aeby 2006 ) to describe lesions facilitates regional comparisons of coral disease.

Hawai'i

Cause-specific temporal and spatial trends in green sea turtle strandings in the Hawaiian Archipelago (1982-2003)

We investigated cause-specific temporal and spatial trends in sea turtle strandings in the Hawaiian Archipelago. Five species of sea turtle were recorded in 3,861 strandings over a 22-year period (1982–2003). Green turtles comprised 97% of these strandings with size and gender composition reflecting the demographic structure of the resident green turtle population and relative green turtle abundance in Hawaiian waters. The cause of strandings was determined by necropsy based on a complete gross external and internal examination. Totally 75% of the 3,732 green turtle strandings were from Oahu where strandings occur year-round. The most common known cause of the green turtle strandings was the tumour-forming disease, fibropapillomatosis (28%) followed by hook-and-line fishing gear-induced trauma (7%), gillnet fishing gear-induced trauma (5%), boat strike (2.5%), and shark attack (2.7%). Miscellaneous causes comprised 5.4% of strandings whereas 49% of green turtle strandings could not be attributed to any known cause. Green turtle strandings attributable to boat strike were more likely from Kauai and Oahu while fibropapilloma strandings were more likely from Oahu and Maui. Hook-and-line gear strandings were more likely from Oahu due to higher per capita inshore fishing effort. The specific mortality rate (conditional probability) for fibropapillomatosis was 88%, 69% for gillnet gear and 52% for hook-and-line gear. The probability of a dead green turtle stranding increased from 1982 but levelled off by the mid-1990s. The declining mortality risk was because the prevalence and severity of fibropapillomatosis has decreased recently and so has the mortality risk attributable to gillnet gear. Despite exposure to disease and inshore fishing gears, the Hawaiian green turtle stock continues to recover following protection since the late 1970s. Nevertheless, measures to reduce incidental capture of sea turtles in coastal Hawaiian fisheries would be prudent, especially since strandings attributable to hook-and-line fishing gear have increased steadily since 1982.

Hawai'i

Biomedical and veterinary science can increase our understanding of coral disease

A balanced approach to coral disease investigation is critical for understanding the global decline of corals. Such an approach should involve the proper use of biomedical concepts, tools, and terminology to address confusion and promote clarity in the coral disease literature. Investigating disease in corals should follow a logical series of steps including identification of disease, systematic morphologic descriptions of lesions at the gross and cellular levels, measurement of health indices, and experiments to understand disease pathogenesis and the complex interactions between host, pathogen, and the environment. This model for disease investigation is widely accepted in the medical, veterinary and invertebrate pathology disciplines. We present standard biomedical rationale behind the detection, description, and naming of diseases and offer examples of the application of Koch's postulates to elucidate the etiology of some infectious diseases. Basic epidemiologic concepts are introduced to help investigators think systematically about the cause(s) of complex diseases. A major goal of disease investigation in corals and other organisms is to gather data that will enable the establishment of standardized case definitions to distinguish among diseases. Concepts and facts amassed from empirical studies over the centuries by medical and veterinary pathologists have standardized disease investigation and are invaluable to coral researchers because of the robust comparisons they enable; examples of these are given throughout this paper. Arguments over whether coral diseases are caused by primary versus opportunistic pathogens reflect the lack of data available to prove or refute such hypotheses and emphasize the need for coral disease investigations that focus on: characterizing the normal microbiota and physiology of the healthy host; defining ecological interactions within the microbial community associated with the host; and investigating host immunity, host-agent interactions, pathology, pathogenesis, and factors that promote the pathogenicity of the causative agent(s) of disease.

Journal of Experimental Marine Biology and Ecology

A new species of Henneguya (myxozoa) in the big-eyed scad (Selar crumenophthalmus) from Hawaii

We describe a new myxozoan, Henneguya akule n. sp., infecting the carangid fish Selar crumenophthalmus in Hawaii. Spores were found only in the aortic bulb, characterized by elliptical capsule with 2 tails, and pyriform polar capsules that angled toward the anterior end of the spore. Polar filaments had 3–4 coils. Parasites were present in apparently healthy fishes and caused no evident gross pathology. On microscopy, parasites evinced a mild inflammatory response in the host characterized by accumulations of eosinophilic fibrillar material around spores and a mononuclear infiltrate in the adventitia of the bulbus arteriosus. Overall prevalence was 20%, and prevalence between 2001 and 2006 ranged from 12 to 27%, but did not differ significantly between years. In contrast, prevalence of infection was highest in south-central Oahu. There was no relationship between infection status and body condition or gender of fish, and infection was absent in the smallest and largest fishes. Phylogenetically, H. akule n. sp. is most closely related to other Henneguya species infecting the heart of marine fishes based on ribosomal DNA analysis. This is the first documentation of a myxozoan parasite in marine fishes from Hawaii.

Hawai'i

Distribution and morphology of growth anomalies in Acropora from the Indo-Pacific

We assessed the distribution and prevalence of growth anomalies (GAs) in Acropora from French Frigate Shoals (Hawaii, USA), Johnston Atoll and Tutuila (American Samoa), developed a nomenclature for gross morphology, characterized GAs at the cellular level and obtained preliminary indices of their spatial patterns and progression within coral colonies. Acropora GAs were found in all 3 regions, but the distribution, variety and prevalence of Acropora GAs was highest in American Samoa. GAs were grouped into 7 gross morphologies (exophytic, bosselated, crateriform, nodular, vermiform, fimbriate or annular). On histology, GAs consisted of hyperplastic basal body wall (calicodermis, mesoglea and gastrodermis apposed to skeleton) with 3 distinct patterns of necrosis. There was no evidence of anaplasia or mitotic figures (common but not necessarily required morphologic indicators of neoplasia). Compared to normal tissues, GAs had significantly fewer polyps, zooxanthellae within the gastrodermis of the coenenchyme, mesenterial filaments and gonads but significantly more necrosis. On 2 colonies with GAs monitored at 2 points over 11 mo, numbers of GAs per colony increased from 0.9 to 3 times the original number seen, and significant clustering of GAs occurred within colonies. The evidence of GAs being true neoplasias (tumors) is mixed, so a cautionary approach is urged in use of morphologic terminology.

Hawai'i

Coral and crustose coralline algae disease on the reefs of American Samoa

Surveys for lesions in corals were conducted at seven sites around Tutuila in June 2004 and January 2005. The objectives of the study were to document the distribution and prevalence of disease in the major genera of corals and crustose coralline algae, systematically describe gross and microscopic morphology of lesions in reef corals and determine whether there are seasonal differences in prevalence of disease. We documented 12 different coral disease states from the reefs of Tutuila and two diseases of crustose coralline algae (CCA). Acropora white syndrome, Acropora growth anomalies and coralline lethal orange disease were the most common diseases on the reefs of Tutuila. No seasonal differences were found in overall prevalence of coral or abundance of CCA disease. Histological analyses of coral lesions revealed that microscopic changes in tissues can be used to distinguish tissue loss due to trauma from changes due to disease, detect micro-organisms associated with certain types of discolorations and found that hyperplasia of the basal body wall was the hallmark microscopic appearance of Acropora growth anomalies regardless of gross morphology of tumors.

Conference Paper

Translocation and disease monitoring of wild laysan ducks

The Laysan duck (Anas laysanensis), also known as the Laysan teal because of its small size, is a critically endangered waterfowl species that once occurred widely across the Hawaiian Archipelago. For the past 150 years, however, it was restricted to a single population on Laysan, a 4-square-kilometer (1.5-square-mile) island with a hypersaline shallow lake. Laysan is part of the Hawaiian Islands National Wildlife Refuge in Papahanaumokuakea Marine National Monument.

Hawaii

Molecular phylogeny of Babesia poelea from brown boobies ( Sula leucogaster ) from Johnston Atoll, Central Pacific

The phylogenetic relationship of avian Babesia with other piroplasms remains unclear, mainly because of a lack of objective criteria such as molecular phylogenetics. In this study, our objective was to sequence the entire 18S, ITS-1, 5.8S, and ITS-2 regions of the rRNA gene and partial ß-tubulin gene of B. poelea , first described from brown boobies ( Sula leucogaster ) from the central Pacific, and compare them to those of other piroplasms. Phylogenetic analyses of the entire 18S rRNA gene sequence revealed that B. poelea belonged to the clade of piroplasms previously detected in humans, domestic dogs, and wild ungulates in the western United States. The entire ITS-1, 5.8S, ITS-2, and partial ß-tubulin gene sequence shared conserved regions with previously described Babesia and Theileria species. The intron of the ß-tubulin gene was 45 bp. This is the first molecular characterization of an avian piroplasm.

Johnston Atoll National Wildlife Refuge

Characterizing lesions in corals from American Samoa

The study of coral disease has suffered from an absence of systematic approaches that are commonly used to determine causes of diseases in animals. There is a critical need to develop a standardized and portable nomenclature for coral lesions in the field and to incorporate more commonly available biomedical tools in coral disease surveys to determine the potential causes of lesions in corals. We characterized lesions in corals from American Samoa based on gross and microscopic morphology and classified them as discoloration, growth anomalies, or tissue loss. The most common microscopic finding in corals manifesting discoloration was the depletion of zooxanthellae, followed by necrosis, sometimes associated with invasive algae or fungi. The most common microscopic lesion in corals manifesting tissue loss was cell necrosis often associated with algae, fungi, or protozoa. Corals with growth anomaly had microscopic evidence of hyperplasia of gastrovascular canals, followed by necrosis associated with algae or metazoa (polychaete worms). Several species of apparently normal corals also had microscopic changes, including the presence of bacterial aggregates or crustacea in tissues. A single type of gross lesion (e.g., discoloration) could have different microscopic manifestations. This phenomenon underlines the importance of using microscopy to provide a more systematic description of coral lesions and to detect potential pathogens associated with these lesions.

Coral Reefs