Marilyn Cruz

Evidence for regular ongoing introductions of mosquito disease vectors into the Galápagos Islands

Wildlife on isolated oceanic islands is highly susceptible to the introduction of pathogens. The recent establishment in the Galápagos Islands of the mosquito Culex quinquefasciatus, a vector for diseases such as avian malaria and West Nile fever, is considered a serious risk factor for the archipelagos endemic fauna. Here we present evidence from the monitoring of aeroplanes and genetic analysis that C. quinquefasciatus is regularly introduced via aircraft into the Galápagos Archipelago. Genetic population structure and admixture analysis demonstrates that these mosquitoes breed with, and integrate successfully into, already-established populations of C. quinquefasciatus in the Galápagos, and that there is ongoing movement of mosquitoes between islands. Tourist cruise boats and inter-island boat services are the most likely mechanism for transporting Culex mosquitoes between islands. Such anthropogenic mosquito movements increase the risk of the introduction of mosquito-borne diseases novel to Galápagos and their subsequent widespread dissemination across the archipelago. Failure to implement and maintain measures to prevent the human-assisted transport of mosquitoes to and among the islands could have catastrophic consequences for the endemic wildlife of Galápagos.

Exposure to Toxoplasma gondii in Galapagos Penguins (Spheniscus mendiculus) and Flightless Cormorants (Phalacrocorax harrisi ) in the Galapagos Islands, Ecuador

Toxoplasma gondii is one of the most common protozoan parasites of humans and warm-blooded animals. Members of the family Felidae are the only definitive hosts of this parasite and, thus, important in the epidemiology of the disease. Previous studies on Pacific islands have found T. gondii infections in a number of avian species where domestic cats (Felis catus) have been introduced. Little is known about T. gondii in the Galapagos Islands, although introduced domestic cats in the archipelago are known to be T. gondii antibody-positive. In this study, we quantified prevalence of antibody to T. gondii in two threatened avian marine species, Galapagos Penguins (Spheniscus mendiculus) and Flightless Cormorants (Phalacrocorax harrisi), and tested the hypothesis that this parasite is more prevalent on Isabela Island (with cats) than on Fernandina Island (without cats). Overall, antibody prevalence was 2.3% in both Galapagos Penguins and Flightless Cormorants from samples collected during 2003–2005, and in 2008. In Galapagos Penguins (n=298), a significantly higher antibody prevalence was found in penguins on Fernandina Island (free of cats) than on Isabela Island (with cats; Fishers exact test; P=0.02). In Flightless Cormorants (n=258), there was a higher antibody prevalence in cormorants living on Isabela than on Fernandina, although this difference was not statistically significant (Fishers; P=0.19). This study is the first to show exposure to T. gondii in endemic avian species in the Galapagos Islands, providing evidence for disease-related risks associated with the feral cat population in the archipelago. We provide possible explanations for these findings and recommendations for future studies towards a better understanding of the epidemiology of T. gondii in the Galapagos Islands.

Immune activity, body condition and human-associated environmental impacts in a wild marine mammal.

Within individuals, immunity may compete with other life history traits for resources, such as energy and protein, and the damage caused by immunopathology can sometimes outweigh the protective benefits that immune responses confer. However, our understanding of the costs of immunity in the wild and how they relate to the myriad energetic demands on free-ranging organisms is limited. The endangered Galapagos sea lion (Zalophus wollebaeki) is threatened simultaneously by disease from domestic animals and rapid changes in food availability driven by unpredictable environmental variation. We made use of this unique ecology to investigate the relationship between changes in immune activity and changes in body condition. We found that during the first three months of life, changes in antibody concentration were negatively correlated with changes in mass per unit length, skinfold thickness and serum albumin concentration, but only in a sea lion colony exposed to anthropogenic environmental impacts. It has previously been shown that changes in antibody concentration during early Galapagos sea lion development were higher in a colony exposed to anthropogenic environmental impacts than in a control colony. This study allows for the possibility that these relatively large changes in antibody concentration are associated with negative impacts on fitness through an effect on body condition. Our findings suggest that energy availability and the degree of plasticity in immune investment may influence disease risk in natural populations synergistically, through a trade-off between investment in immunity and resistance to starvation. The relative benefits of such investments may change quickly and unpredictably, which allows for the possibility that individuals fine-tune their investment strategies in response to changes in environmental conditions. In addition, our results suggest that anthropogenic environmental impacts may impose subtle energetic costs on individuals, which could contribute to population declines, especially in times of energy shortage.

Causes of mortality of wild birds submitted to the Charles Darwin Research Station, Santa Cruz, Galapagos, Ecuador from 2002-2004.

Necropsy findings were reviewed from wild birds submitted to the Charles Darwin Research Station, Santa Cruz Island, Galápagos Archipelago between 2004 and 2006. One hundred and ninety cases from 27 different species were submitted, and 178 of these cases were evaluated grossly or histologically. Trauma and trauma-related deaths (n=141) dominated necropsy submissions. Infectious causes of avian mortality included myiasis due to Philornis sp. (n=6), avian pox (n=1), and schistosomosis (n=1).

Host selection and parasite infection in Aedes taeniorhynchus, endemic disease vector in the Galapagos Islands

Host selection in blood-sucking arthropods has important evolutionary and ecological implications for the transmission dynamics, distribution and host-specificity of the parasites they transmit. The black salt-marsh mosquito (Aedes taeniorhynchus Wiedemann) is distributed throughout tropical to temperate coastal zones in the Americas, and continental populations are primarily mammalphilic. It is the only indigenous mosquito in the Galápagos Islands, having colonised the archipelago around 200,000 years ago, potentially adapting its host selection, and in the process, altering the dynamics of vector mediated pathogen interactions in the archipelago. Here, we use blood-meal analysis and PCR-based parasite screening approach to determine the blood-feeding patterns of A. taeniorhynchus in the Galápagos Islands and identify potential parasite transmission with which this mosquito could be involved. Our results show that A. taeniorhynchus feeds equally on mammals and reptiles, and only one avian sample was observed in 190 successful PCR amplifications from blood meals. However, we detected endemic filarial worms and Haemoproteus parasites known to infect various Galápagos bird species in mosquito thoraces, suggesting that feeding on birds must occur at low frequency, and that A. taeniorhynchus may play a role in maintaining some avian vector-borne pathogens, although more work is needed to explore this possibility. We also isolated three different DNA sequences corresponding to hemogregarine parasites of the genus Hepatozoon from mosquito and iguana blood samples, suggesting that more than one species of Hepatozoon parasites are present in Galápagos. Phylogenetic analysis of Hepatozoon 18sRNA sequences indicates that A. taeniorhynchus may have facilitated a recent breakdown in host-species association of formerly isolated Hepatozoon spp. infecting the reptile populations in the Galápagos Islands.

COMPARISON OF BLOOD VALUES AND HEALTH STATUS OF FLOREANA MOCKINGBIRDS (MIMUS TRIFASCIATUS) ON THE ISLANDS OF CHAMPION AND GARDNER-BY-FLOREANA, GALÁPAGOS ISLANDS

The Floreana Mockingbird (Mimus trifasciatus) is one of the rarest bird species in the world, with an estimated 550 individuals remaining on two rocky islets off the coast of Floreana, Galápagos, Ecuador, from which the main population was extirpated more than 100 yr ago. Because they have been listed in critical danger of extinction, a plan to reintroduce this species to Floreana has been initiated. Determining the health status of the source mockingbird populations is a top priority within the reintroduction plan. We report the health status, over the course of 4 yr, of 75 Floreana Mockingbirds on Champion Island and 160 Floreana Mockingbirds on Gardner-by-Floreana, based on physical examinations, hematology, hemolysis–hemagglutination assay, exposure to selected infectious disease agents, and ecto- and endoparasite counts. Birds on Gardner-by-Floreana had higher body condition index scores, packed cell volumes, total solids, and lymphocyte counts. Additionally, Gardner-by-Floreana birds had lower heterophil counts, eosinophil counts, and heterophil:lymphocyte ratios. No Chlamydophila psittaci DNA or antibodies to paramyxovirus-I, adenovirus-II, or Mycoplasma gallisepticum were found in any of the mockingbirds tested. Ectoparasites were present on birds from both islands, although species varied between islands. A coccidian species was found in eight of the 45 fecal samples from birds on Gardner-by-Floreana, but none of 33 birds examined from Champion. Birds on Gardner-by-Floreana were classified as healthier than those on Champion based on clinical and laboratory findings. These health data will be analyzed in conjunction with genetics, population structure, and disease presence on Floreana for developing recommendations for the Floreana Mockingbird reintroduction plan.

Seroprevalence of Malarial Antibodies in Galapagos Penguins (Spheniscus mendiculus)

Abstract:  A parasite species of the genus Plasmodium has recently been documented in the endangered Galapagos penguin (Spheniscus mendiculus). Avian malaria causes high mortality in several species after initial exposure and there is great concern for the conservation of the endemic Galapagos penguin. Using a Plasmodium spp. circumsporozoite protein antigen, we standardized an enzyme-linked immunosorbent assay to test the level of exposure in this small population, as indicated by seroprevalence. Sera from adult and juvenile Galapagos penguins collected between 2004 and 2009 on the Galapagos archipelago were tested for the presence of anti-Plasmodium spp. antibodies. Penguins were also tested for the prevalence of avian malaria parasite DNA using a polymerase chain reaction (PCR) screening. Total seroprevalence of malarial antibodies in this sample group was 97.2%, which suggests high exposure to the parasite and low Plasmodium-induced mortality. However, total prevalence of Plasmodium parasite DNA by PCR screening was 9.2%, and this suggests that parasite prevalence may be under-detected through PCR screening. Multiple detection methods may be necessary to measure the real extent of Plasmodium exposure on the archipelago.

Seasonal effects and fine-scale population dynamics of Aedes taeniorhynchus, a major disease vector in the Galapagos Islands

Characterization of the fine‐scale population dynamics of the mosquito Aedes taeniorhynchus is needed to improve our understanding of its role as a disease vector in the Galapagos Islands. We used microsatellite data to assess the genetic structure of coastal and highland mosquito populations and patterns of gene flow between the two habitats through time on Santa Cruz Island. In addition, we assessed possible associations of mosquito abundance and genetic diversity with environmental variables. The coastal and highland mosquito populations were highly differentiated from each other all year round, with some gene flow detected only during periods of increased precipitation. The results support the hypothesis that selection arising from ecological differences between habitats is driving adaptation and divergence in A. taeniorhynchus, and maintaining long‐term genetic differentiation of the populations against gene flow. The highland and lowland populations may constitute an example of incipient speciation in progress. Highland populations were characterized by lower observed heterozygosity and allelic richness, suggesting a founder effect and/or lower breeding site availability in the highlands. A lack of reduction in genetic diversity over time in highland populations suggests that they survive dry periods as dormant eggs. Association between mosquito abundance and precipitation was strong in the highlands, whereas tide height was the main factor affecting mosquito abundance on the coast. Our findings suggests differences in the infection dynamics of mosquito‐borne parasites in the highlands compared to the coast, and a higher risk of mosquito‐driven disease spread across these habitats during periods of increased precipitation.

Adaptation, isolation by distance and human-mediated transport determine patterns of gene flow among populations of the disease vector Aedes taeniorhynchus in the Galapagos Islands

The black salt-marsh mosquito (Aedes taeniorhynchus) is the only native mosquito in the Galapagos Islands and potentially a major disease vector for Galapagos wildlife. Little is known about its population structure, or how its dynamics may be influenced by human presence in the archipelago. We used microsatellite data to assess the structure and patterns of A. taeniorhynchus gene flow among and within islands, to identify potential barriers to mosquito dispersal, and to investigate human-aided transport of mosquitoes across the archipelago. Our results show that inter-island migration of A. taeniorhynchus occurs frequently on an isolation by distance basis. High levels of inter-island migration were detected amongst the major ports of the archipelago, strongly suggesting the occurrence of human-aided transport of mosquitoes among islands, underlining the need for strict control measures to avoid the transport of disease vectors between islands. The prevalence of filarial nematode infection in Galapagos flightless cormorants is correlated with the population structure and migration patterns of A. taeniorhynchus, suggesting that A. taeniorhynchus is an important vector of this arthropod-borne parasite in the Galapagos Islands. Therefore mosquito population structure in Galapagos may have the potential to influence mosquito-borne parasite population dynamics, and the subsequent impacts of such pathogens on their host species in the islands.

Context-dependent associations between heterozygosity and immune variation in a wild carnivore

BackgroundA multitude of correlations between heterozygosity and fitness proxies associated with disease have been reported from wild populations, but the genetic basis of these associations is unresolved. We used a longitudinal dataset on wild Galapagos sea lions (Zalophus wollebaeki) to develop a relatively new perspective on this problem, by testing for associations between heterozygosity and immune variation across age classes and between ecological contexts.ResultsHomozygosity by locus was negatively correlated with serum immunoglobulin G production in pups (0–3 months of age), suggesting that reduced genetic diversity has a detrimental influence on the early development of immune defence in the Galapagos sea lion. In addition, homozygosity by locus was positively correlated with total circulating leukocyte concentration in juveniles (6–24 months of age), but only in a colony subject to the anthropogenic environmental impacts of development, pollution and introduced species, which suggests that reduced genetic diversity influences mature immune system activity in circumstances of high antigen exposure.ConclusionsThese findings demonstrate the environmental context-dependency of the phenotypic expression of immune variation, which is implicit in the theory of ecoimmunology, but which has been rarely demonstrated in the wild. They also indicate that heterozygosity may be linked to the maintenance of heterogeneity in mammalian immune system development and response to infection, adding to the body of evidence on the nature of the mechanistic link between heterozygosity and fitness.