Julie C. Ellis

Victims or vectors: a survey of marine vertebrate zoonoses from coastal waters of the Northwest Atlantic

Surveillance of zoonotic pathogens in marine birds and mammals in the Northwest Atlantic revealed a diversity of zoonotic agents. We found amplicons to sequences from Brucella spp., Leptospira spp., Giardia spp. and Cryptosporidium spp. in both marine mammals and birds. Avian influenza was detected in a harp seal and a herring gull. Routine aerobic and anaerobic culture showed a broad range of bacteria resistant to multiple antibiotics. Of 1460 isolates, 797 were tested for resistance, and 468 were resistant to one or more anti-microbials. 73% (341/468) were resistant to 1-4 drugs and 27% (128/468) resistant to 5-13 drugs. The high prevalence of resistance suggests that many of these isolates could have been acquired from medical and agricultural sources and inter-microbial gene transfer. Combining birds and mammals, 45% (63/141) of stranded and 8% (2/26) of by-caught animals in this study exhibited histopathological and/or gross pathological findings associated with the presence of these pathogens. Our findings indicate that marine mammals and birds in the Northwest Atlantic are reservoirs for potentially zoonotic pathogens, which they may transmit to beachgoers, fishermen and wildlife health personnel. Conversely, zoonotic pathogens found in marine vertebrates may have been acquired via contamination of coastal waters by sewage, run-off and agricultural and medical waste. In either case these animals are not limited by political boundaries and are therefore important indicators of regional and global ocean health.

Molecular characterization of Giardia intestinalis haplotypes in marine animals: variation and zoonotic potential

Giardia intestinalis is a microbial eukaryotic parasite that causes diarrheal disease in humans and other vertebrates worldwide. The negative effect on quality of life and economics caused by G. intestinalis may be increased by its potential status as a zoonosis, or a disease that can be transmitted from animals to humans. The zoonotic potential of G. intestinalis has been implied for over 2 decades, with human-infecting genotypes (belonging to the 2 major subgroups, Assemblages A and B) occurring in wildlife and domesticated animals. There are recent reports of G. intestinalis in shellfish, seals, sea lions and whales, suggesting that marine animals are also potential reservoirs of human disease. However, the prevalence, genetic diversity and effect of G. intestinalis in marine environments and the role that marine animals play in transmission of this parasite to humans are relatively unexplored. Here, we provide the first thorough molecular characterization of G. intestinalis in marine vertebrates. Using a multi-locus sequencing approach, we identify human-infecting G. intestinalis haplotypes of both Assemblages A and B in the fecal material of dolphins, porpoises, seals, herring gulls Larus argentatus, common eiders Somateria mollissima and a thresher shark Alopias vulpinus. Our results indicate that G. intestinalis is prevalent in marine ecosystems, and a wide range of marine hosts capable of harboring zoonotic forms of this parasite exist. The presence of G. intestinalis in marine ecosystems raises concerns about how this disease might be transmitted among different host species.

Marine Foraging Birds As Bioindicators of Mercury in the Gulf of Maine

From existing databases, we compiled and evaluated 604 total mercury (Hg) levels in the eggs and blood of 17 species of marine foraging birds from 35 Gulf of Maine islands to provide baseline data and to determine the best tissue, age class, and species for future biomonitoring. While mean Hg levels in most species did not exceed adverse effects thresholds, levels in some individual eggs did; for all species arithmetic mean egg Hg levels ranged from 0.04 to 0.62 (μg/g, wet weight). Piscivorous birds had higher Hg levels than invertivores. Leach’s storm-petrel (Oceanodroma leucorhoa), razorbill (Alca torda), and black guillemot (Cepphus grylle) adult blood and egg Hg levels were higher than other species. Our results indicate that adult blood is preferable to chick blood for detecting long-term temporal trends because adult levels are higher and not confounded by metabolic effects. However, since we found that eggs and adult blood are comparable indicators of methylmercury bioavailability, we determined that eggs are the preferred tissue for long-term Hg monitoring because the relative ease in collecting eggs ensures consistent and robust datasets. We suggest specific sampling methods, and based on our results demonstrate that common eider (Somateria mollissima), Leach’s storm-petrel, double-crested cormorant, and black guillemot are the most effective bioindicators of Hg of the Gulf of Maine.

Occurrence and patterns of antibiotic resistance in vertebrates off the Northeastern United States coast

The prevalence of antibiotic-resistant bacteria in the marine environment is a growing concern, but the degree to which marine mammals, seabirds and fish harbor these organisms is not well documented. This project sought to identify the occurrence and patterns of antibiotic resistance in bacteria isolated from vertebrates of coastal waters in the northeastern United States. Four hundred and seventy-two isolates of clinical interest were tested for resistance to a suite of 16 antibiotics. Fifty-eight percent were resistant to at least one antibiotic, while 43% were resistant to multiple antibiotics. A multiple antibiotic resistance index value >or=0.2 was observed in 38% of the resistant pathogens, suggesting exposure of the animals to bacteria from significantly contaminated sites. Groups of antibiotics were identified for which bacterial resistance commonly co-occurred. Antibiotic resistance was more widespread in bacteria isolated from seabirds than marine mammals, and was more widespread in stranded or bycaught marine mammals than live marine mammals. Structuring of resistance patterns based on sample type (live/stranded/bycaught) but not animal group (mammal/bird/fish) was observed. These data indicate that antibiotic resistance is widespread in marine vertebrates, and they may be important reservoirs of antibiotic-resistant bacteria in the marine environment.

Functional Metagenomics Reveals Previously Unrecognized Diversity of Antibiotic Resistance Genes in Gulls

Wildlife may facilitate the spread of antibiotic resistance (AR) between human-dominated habitats and the surrounding environment. Here, we use functional metagenomics to survey the diversity and genomic context of AR genes in gulls. Using this approach, we found a variety of AR genes not previously detected in gulls and wildlife, including class A and C β-lactamases as well as six tetracycline resistance gene types. An analysis of the flanking sequences indicates that most of these genes are present in Enterobacteriaceae and various Gram-positive bacteria. In addition to finding known gene types, we detected 31 previously undescribed AR genes. These undescribed genes include one most similar to an uncharacterized gene in Verrucomicrobium and another to a putative DNA repair protein in Lactobacillus. Overall, the study more than doubled the number of clinically relevant AR gene types known to be carried by gulls or by wildlife in general. Together with the propensity of gulls to visit human-dominated habitats, this high diversity of AR gene types suggests that gulls could facilitate the spread of AR.

Nest Attributes, Aggression, and Breeding Success of Gulls in Single and Mixed Species Subcolonies

Abstract We investigated attributes of nests, aggressive interactions, and reproductive success in mixed and single species subcolonies of Great Black-backed Gulls (Larus marinus) and Herring Gulls (L. argentatus) on Appledore Island, Maine. Distances to the nearest neighboring nest were smaller for L. argentatus than L. marinus, with greatest distances between nests at edges of subcolonies in both species. More L. argentatus nests had natural screens (vegetation or rock >30 cm tall) adjacent to them than did L. marinus nests, but screen presence did not differ among nest positions within the colony. Clutch size did not differ between species; however, hatching success (number of chicks hatched per nest) was higher in L. argentatus than L. marinus. Fledging success (number of chicks fledged per nest) of L. marinus was greater at nests with heterospecific neighbors, whereas the opposite was true for L. argentatus. For both species, the frequency of aggressive interactions was lower at nests with L. argentatus neighbors. Overall, L. marinus nesting near L. argentatus experienced less aggression and greater reproductive success than those nesting among conspecifics, where intraspecific aggression was relatively high. L. argentatus nesting near L. marinus experienced more aggression and lower reproductive success than those nesting among conspecifics, where intraspecific aggression was relatively low. The costs and benefits of nesting in mixed species colonies may depend on the relative size and aggressiveness of the heterospecifics. Interactions with L. marinus in mixed species colonies may be contributing to the current declines of L. argentatus throughout New England.

Impact of Raccoons on Breeding Success in Large Colonies of Great Black-backed Gulls and Herring Gulls

Abstract For most ground-nesting seabirds, offshore islands usually serve as refuges from mammalian predators, however, introduced predators pose a substantial threat to many species. In New England, thousands of Herring Gulls (Larus argentatus) and Great Black-backed Gulls (L. marinus) breed on hundreds of coastal islands. The Isles of Shoals (New Hampshire/Maine) archipelago has some of the largest colonies of both gull species in New England. In 2004, we noted unusually large numbers of failed nests on Appledore Island, Maine and hypothesized that Raccoons (Procyon lotor) were the cause. Therefore, we quantified egg and chick mortality and documented the presence of Raccoons. We trapped Raccoons on Appledore and Smuttynose islands and conducted nest surveys on these and two other islands in the Isles of Shoals. Surveys of marked nests on Appledore Island revealed high levels of egg and chick mortality in early June 2004; by late June 58% of nests were empty. We also found that on Appledore and Smuttynose, where Raccoons were observed, the odds that a nest would be empty were 17 times greater than on Lunging or Duck Islands where no Raccoons were observed. During 2004, eleven Raccoons were trapped off of Appledore and Smuttynose Islands. A survey of nests in 2005 showed that gull breeding success increased significantly on Appledore and Smuttynose islands, whereas there was no significant change in the relatively high ratio of chicks to nests from 2004 to 2005 on Lunging Island where Raccoons were not observed. Thus, we concluded that the presence of a few Raccoons on Appledore and Smuttynose islands was sufficient to cause substantial breeding failure in large colonies of larids.

Cyclic Avian Mass Mortality in the Northeastern United States Is Associated with a Novel Orthomyxovirus

ABSTRACT Since 1998, cyclic mortality events in common eiders (Somateria mollissima), numbering in the hundreds to thousands of dead birds, have been documented along the coast of Cape Cod, MA, USA. Although longitudinal disease investigations have uncovered potential contributing factors responsible for these outbreaks, detecting a primary etiological agent has proven enigmatic. Here, we identify a novel orthomyxovirus, tentatively named Wellfleet Bay virus (WFBV), as a potential causative agent of these outbreaks. Genomic analysis of WFBV revealed that it is most closely related to members of the Quaranjavirus genus within the family Orthomyxoviridae. Similar to other members of the genus, WFBV contains an alphabaculovirus gp64-like glycoprotein that was demonstrated to have fusion activity; this also tentatively suggests that ticks (and/or insects) may vector the virus in nature. However, in addition to the six RNA segments encoding the prototypical structural proteins identified in other quaranjaviruses, a previously unknown RNA segment (segment 7) encoding a novel protein designated VP7 was discovered in WFBV. Although WFBV shows low to moderate levels of sequence similarity to Quaranfil virus and Johnston Atoll virus, the original members of the Quaranjavirus genus, additional antigenic and genetic analyses demonstrated that it is closely related to the recently identified Cygnet River virus (CyRV) from South Australia, suggesting that WFBV and CyRV may be geographic variants of the same virus. Although the identification of WFBV in part may resolve the enigma of these mass mortality events, the details of the ecology and epidemiology of the virus remain to be determined. IMPORTANCE The emergence or reemergence of viral pathogens resulting in large-scale outbreaks of disease in humans and/or animals is one of the most important challenges facing biomedicine. For example, understanding how orthomyxoviruses such as novel influenza A virus reassortants and/or mutants emerge to cause epidemic or pandemic disease is at the forefront of current global health concerns. Here, we describe the emergence of a novel orthomyxovirus, Wellfleet Bay virus (WFBV), which has been associated with cyclic large-scale bird die-offs in the northeastern United States. This initial characterization study provides a foundation for further research into the evolution, epidemiology, and ecology of newly emerging orthomyxoviruses, such as WFBV, and their potential impacts on animal and/or human health.

Bathing birds bias β-diversity: Frequent dispersal by gulls homogenizes fauna in a rock-pool metacommunity

Metacommunity theory generally predicts that regional dispersal of organisms among local habitat patches should influence spatial patterns of species diversity. In particular, increased dispersal rates are generally expected to increase local (alpha) diversity, yet homogenize local communities across the region (decreasing beta-diversity), resulting in no change in regional (gamma) diversity. Although the effect of dispersal on alpha-diversity has garnered much experimental attention, the influence of dispersal rates on diversity at larger spatial scales (beta and gamma) is poorly understood. Furthermore, these theoretical predictions are not well tested in the field, where other environmental factors (e.g., habitat size, resource density) likely also influence species diversity. Here, we used a system of freshwater rock pools on Appledore Island, Maine, USA, to test the effects of dispersal rate on species diversity in metacommunities. The pools exist in clusters (metacommunities) that experience different levels of dispersal imposed by gulls (Larus spp.), which we show to be frequent passive dispersers of rock-pool invertebrates. Although previous research has suggested that waterbirds may disperse aquatic invertebrates, our study is the first to quantify the rate at which such dispersal occurs and determine its effects on species diversity. In accordance with theory, we found that metacommunities experiencing higher dispersal rates had significantly more homogeneous local communities (reduced beta-diversity) and that gamma-diversity was not influenced by dispersal rate. Contrary to theoretical predictions, however, alpha-diversity in the rock pools was not significantly influenced by dispersal. Rather, local diversity was significantly positively related to local habitat size, and both alpha- and gamma-diversity were influenced by the physicochemical environment of the pools. These results provide an important field test of metacommunity theory, highlighting how local and regional factors interact to drive patterns of species diversity in metacommunities, and demonstrate that waterbirds are indeed important dispersal vectors for aquatic invertebrates.

Pilot Study of Antimicrobial-Resistant Escherichia coli in Herring Gulls (Larus argentatus) and Wastewater in the Northeastern United States

Abstract Wildlife may be an important reservoir of antibiotic-resistant bacteria and resistance genes. In this pilot study, the prevalence and patterns of antimicrobial resistance in Escherichia coli cultured from wild herring gull (Larus argentatus) feces and human wastewater at Cape Cod, Massachusetts, USA, was compared. Antimicrobial susceptibility was tested using Kirby–Bauer disk diffusion with seven antimicrobial agents. A high proportion of antimicrobial agent-resistant E. coli isolates (59.2%) were detected in wastewater samples compared with a lower prevalence of 17.5% in gull feces. In addition, there was a large proportion of isolates with intermediate susceptibility (93.0%) in gull feces. Although similar resistance patterns and shared resistance genes suggest possible wastewater contamination of the local environment, the relatively low frequency of resistance and high prevalence of intermediate susceptibility detected in E. coli cultured from gull feces depict a complex model of antimicrobial resistance among E. coli strains of wildlife origin.