Expansion of an exotic species and concomitant disease outbreaks: pigeon paramyxovirus in free-ranging Eurasian collared doves.

Eurasian collared doves (Streptopelia decaocto) have expanded their range across the United States since their introduction several decades ago. Recent mortality events in Eurasian collared doves in Arizona and Montana, USA, during the winter of 2009-2010 were the result of pigeon paramyxovirus (PPMV), a novel disease agent. The first instance of mortality by this emerging infectious disease in this species occurred in Florida in 2001 with subsequent disease events in 2006 and 2008. Full diagnostic necropsies were performed on carcasses from the three states. PPMV was identified by RT-PCR and virus isolation and was sequenced to the VIb genotype of avian paramyxovirus-1 (APMV). Other APMVs are common in a variety of free-ranging birds, but concern is warranted because of the potential for commingling of this species with native birds, virus evolution, and threats to domestic poultry. Improved surveillance for wildlife mortality events and efforts to prevent introduction of non-native animals could reduce the threat of introducing new pathogens.

Mite-filled cyst on a Brown-headed Cowbird (Molothrus ater) in Florida, USA.

A large, partly pedunculated mass on the scapular area of a wild-caught captive Brown-headed Cowbird (Molothrus ater) consisted of a multiloculated keratin cyst inhabited by a new species of harpirhynchid mite (Harpirhynchus quasimodo). The mass did not interfere with flight or behavior. This is the first record of such an infestation of cowbirds in Florida.

Survey for antibodies to infectious bursal disease virus serotype 2 in wild turkeys and Sandhill Cranes of Florida, USA.

Captive-reared Whooping Cranes (Grus americana) released into Florida for the resident reintroduction project experienced unusually high mortality and morbidity during the 1997-98 and 2001-02 release seasons. Exposure to infectious bursal disease virus (IBDV) serotype 2 as evidenced by seroconversion was suspected to be the factor that precipitated these mortality events. Very little is known about the incidence of IBD in wild bird populations. Before this study, natural exposure had not been documented in wild birds of North America having no contact with captive-reared cranes, and the prevalence and transmission mechanisms of the virus in wild birds were unknown. Sentinel chickens (Gallus gallus) monitored on two Whooping Crane release sites in central Florida, USA, during the 2003-04 and 2004-05 release seasons seroconverted, demonstrating natural exposure to IBDV serotype 2. Blood samples collected from Wild Turkeys (Meleagris gallopavo) and Sandhill Cranes (Grus canadensis) in eight of 21 counties in Florida, USA, and one of two counties in southern Georgia, USA, were antibody-positive for IBDV serotype 2, indicating that exposure from wild birds sharing habitat with Whooping Cranes is possible. The presence of this virus in wild birds in these areas is a concern for the resident flock of Whooping Cranes because they nest and raise their chicks in Florida, USA. However, passively transferred antibodies may protect them at this otherwise vulnerable period in their lives.

Effects of methylmercury exposure on the immune function of juvenile common loons (Gavia immer).

We conducted a dose-response laboratory study to quantify the level of exposure to dietary Hg, delivered as methylmercury chloride (CH3HgCl), that is associated with suppressed immune function in captive-reared common loon (Gavia immer) chicks. We used the phytohemagglutinin (PHA) skin test to assess T-lymphocyte function and the sheep red blood cell (SRBC) hemagglutination test to measure antibody-mediated immunity. The PHA stimulation index among chicks receiving dietary Hg treatment did not differ significantly from those of chicks on the control diet (p = 0.15). Total antibody (immunoglobulin [Ig] M [primary antibody] + IgG [secondary response]) production to the SRBC antigen in chicks treated with dietary methylmercury (MeHg), however, was suppressed (p = 0.04) relative to chicks on control diets. Analysis indicated suppression of total Ig production (p = 0.025 with comparisonwise alpha level = 0.017) between control and 0.4 microg Hg/g wet food intake treatment groups. Furthermore, the control group exhibited a higher degree of variability in antibody response compared to the Hg groups, suggesting that in addition to reducing the mean response, Hg treatment reduced the normal variation attributable to other biological factors. We observed bursal lymphoid depletion in chicks receiving the 1.2 microg Hg/g treatment (p = 0.017) and a marginally significant effect (p = 0.025) in chicks receiving the 0.4 microg Hg/g diet. These findings suggest that common loon chick immune systems may be compromised at an ecologically relevant dietary exposure concentration (0.4 microg Hg/g wet wt food intake). We also found that chicks hatched from eggs collected from low-pH lakes exhibited higher levels of lymphoid depletion in bursa tissue relative to chicks hatched from eggs collected from neutral-pH lakes.

Subchronic effects of methylmercury on plasma and organ biochemistries in great egret nestlings.

In recent years, high concentrations of mercury have been found in wading birds in Florida, USA. Great egret (Ardea alba) chicks (2 weeks old) were dosed orally daily with the equivalent of 0, 0.5, or 5 microg/g Hg as methylmercury chloride in the diet for up to 12 weeks. Weakness of the legs or paralysis occurred in all high-dosed birds. Geometric mean blood Hg concentrations were 0.17, 10.3, and 78.5 microg/g (wet wt), respectively. Mercury concentrations for organs (microg/g wet wt), including brain (0.22, 3.4, and 35, respectively), liver (0.34, 15.1, 138, respectively), and kidney (0.28, 8.1, and 120, respectively), increased in a dose-dependent manner. Total glutathione (GSH) peroxidase activity was significantly lower in the plasma, brain, liver, and kidney of the high-dosed group. Plasma aspartate aminotransferase activity increased with mercury treatment, whereas lactate dehydrogenase activity decreased. Four other plasma chemistries were decreased significantly in the high-dosed group and included uric acid, total protein, albumin, and inorganic phosphorus. Lipid peroxidation increased in liver (low and high dose) and brain (high dose). Tissue changes in concentrations of reduced thiols included decreased total thiols and protein-bound thiols in liver, decreased protein-bound thiols in kidney, and increased GSH in kidney and brain. Activities of GSH S-transferase and oxidized glutathione reductase increased in liver. In kidney, GSH S-transferase and glucose-6-phosphate dehydrogenase activities increased with mercury dose. These findings, including apparent compensatory changes, are compared to other Hg studies where oxidative stress was reported in egrets, herons, and diving ducks in the field and mallards in the laboratory.

A historical record of mercury contamination in Southern Florida (USA) as inferred from avian feather tissue.

During the late 1980s, the upper trophic-level biota of the Everglades (FL, USA) was recognized as being highly contaminated with mercury (Hg). However, the timing and pattern of that increase is poorly known, and no information is available about mercury contamination in Everglades wildlife prior to 1974. We measured methylmercury concentrations in feathers of white ibises (n = 33), great egrets (n = 7), anhingas (n = 21), and great blue herons (n = 12) from museum specimens collected from 1910 through 1980 and combined them with more recent feather samples collected from live birds (1985-2000, n = 98, 37, 49, and 7, respectively). We found no evidence of contamination of museum samples with inorganic mercuric preservatives (0.01-0.28% of total Hg in feathers). All species showed relatively low concentrations of mercury through the 1970s (<5 microl/L dry wt for anhingas, ibises, and egrets, <10 microl/L for herons). Samples from all species taken during the 1990s showed a large and significant increase (4-5X) in MeHg concentration. This evidence suggests that most of the increase in Hg deposition during the 20th century in south Florida occurred during the last two to three decades, which is consistent with information about local source deposition. Contamination levels prior to the 1970s appear to have been associated with normal reproduction in these birds, suggesting partial evidence for a threshold of reproductive impairment.

Haemoproteus balearicae and other blood parasites of free-ranging Florida sandhill crane chicks.

We obtained blood smears from 114 Florida sandhill crane (Grus canadensis pratensis) chicks in Osceola and Lake Counties, Florida, USA, during 1998-2000. Leucocytozoon grusi was observed in 11 (10%) chicks; Haemoproteus antigonis was observed in eight (7%) chicks; and three (3%) chicks were infected with Haemoproteus balearicae. One chick infected with H. balearicae suffered from severe anemia (packed cell volume = 13%) and was later found moribund. At necropsy this bird also had severe anemia and damage to the heart possibly due to hypoxia. This is the first report of H. balearicae in free-ranging North American cranes.

Epizootiology of Eustrongylides ignotus in Florida: transmission and development of larvae in intermediate hosts.

Under laboratory conditions, 2 modes of transmission of Eustrongylides ignotus (Nematoda: Dioctophymatoidea) to fish were identified. Eastern mosquitofish (Gambusia holbrooki) became infected after ingestion of either eggs of E. ignotus containing first-stage larvae or aquatic oligochaetes (Limnodrilus hoffmeisteri) containing third-stage larvae of E. ignotus. After removal from the uterus of gravid E. ignotus females and incubation for 17-28 days, depending on temperature, it was found that parasite eggs contained first-stage larvae that were infective to fish and oligochaetes. Larvae developed to the third stage in oligochaetes and were infective to fish 35-77 days postinfection (PI) and when fed to fish, developed to the fourth stage between 127 and 184 days PI. Eggs containing first-stage larvae fed directly to fish developed to the fourth stage between 84 and 105 days PI. The amount of time for development from the undifferentiated egg to the fourth-stage larva was 78-156 days shorter when fish ingested eggs containing first-stage larvae than when fish ingested oligochaetes containing third-stage larvae. Three species of large piscivorous fish, including black crappie (Pomoxis nigromaculatus), largemouth bass (Micropterus salmoides), and warmouth (Lepomis gulosus), were fed mosquitofish containing fourth-stage larvae. At necropsy, live E. ignotus larvae were recovered from all 3 species. Several fish had multiple infections after ingesting > 1 larva, indicating that bioaccumulation of the parasite in the food chain may occur.

Epizootiology of Eustrongylides ignotus in Florida: distribution, density, and natural infections in intermediate hosts.

A total of 63,451 fish, representing 39 species, was collected from 176 foraging sites used by ciconiiform wading birds in peninsular Florida (USA) and examined for larvae of Eustrongylides ignotus. Infected fish were identified from 30 (17%) of the sites, all of which had been altered by human disturbance such as removal of sediment to construct ditches and dikes, improve water flow, or increase storage capacity and had a history of receiving anthropogenic nutrients such as sewage effluent, urban runoff, or agricultural runoff. The mosquitofish (Gambusia holbrooki) and several species of sunfish (Centrarchidae) were the most important intermediate hosts. Infected fish were not collected at any of the unaltered sites. A total of 10,508 oligochaetes (representing 36 species) was identified from 22 sites that had fish infected with E. ignotus and 36 sites where no infected fish were collected. None of the oligochaetes was infected with larvae of E. ignotus. Immature tubificids without hair setae (probably Limnodrilus sp.), Dero digitata, and L. hoffmeisteri were the most abundant oligochaetes at sites where infected fish occurred, making up 78% of the total collected. Compared to unaltered sites, altered sites were characterized by higher mean densities of fish and oligochaetes; surface waters with decreased dissolved oxygen and increased total nitrogen, total phosphorus, and chlorophyll-a; sediments with higher soil oxygen demand and total phosphorus; larger grain sizes; and higher percentage emergent vegetation and grasses.

Nineteenth century mercury: hazard to wading birds and cormorants of the Carson River, Nevada.

Contemporary mercury interest relates to atmospheric deposition, contaminated fish stocks and exposed fish-eating wildlife. The focus is on methylmercury (MeHg) even though most contamination is of inorganic (IoHg) origin. However, IoHg is readily methylated in aquatic systems to become more hazardous to vertebrates. In response to a classic episode of historical (1859-1890) IoHg contamination, we studied fish-eating birds nesting along the lower Carson River, Nevada. Adult double-crested cormorants (Phalacrocorax auritus), snowy egrets (Egretta thula) and black-crowned night-herons (Nycticorax nycticorax) contained very high concentrations of total mercury (THg) in their livers (geo. means 134.8 microg/g wet weight (ww), 43.7 and 13.5, respectively) and kidneys (69.4, 11.1 and 6.1, respectively). Apparently tolerance of these concentrations was possible due to a threshold-dependent demethylation coupled with sequestration of resultant IoHg. Demethylation and sequestration processes also appeared to have reduced the amount of MeHg redistributed to eggs. However, the relatively short time spent by adults in the contaminated area before egg laying was also a factor in lower than expected concentrations of mercury in eggs. Most eggs (100% MeHg) had concentrations below 0.80 microg/g ww, the putative threshold concentration where reproductive problems may be expected; there was no conclusive evidence of mercury-related depressed hatchability. After hatching, the young birds were fed diets by their parents averaging 0.36-1.18 microgMeHg/g ww through fledging. During this four to six week period, accumulated mercury concentrations in the organs of the fledglings were much lower than found in adults, but evidence was detected of toxicity to their immune (spleen, thymus, bursa), detoxicating (liver, kidneys) and nervous systems. Several indications of oxidative stress were also noted in the fledglings and were most apparent in young cormorants containing highest concentrations of mercury. This stress was evidenced by increased thiobarbituric acid-reactive substances, low activities of enzymes related to glutathione metabolism and low levels of reduced thiols, plus an increase in the ratio of oxidized to reduced glutathione. At lower concentrations of mercury, as was found in young egrets, we observed elevated activities of protective hepatic enzymes, which could help reduce oxidative stress. Immune deficiencies and neurological impairment of fledglings may affect survivability when confronted with the stresses of learning to forage and the ability to complete their first migration.

Sarcocystis meningoencephalitis in a northern gannet (Morus bassanus).

Sarcocystis sp. schizonts were found in the cerebellum of a northern gannet (Morus bassanus), exhibiting neurologic signs, found on the Florida (USA) east coast. Based upon molecular characterization of DNA isolated from the brain of the gannet, this Sarcocystis sp. appeared to be closely related, if not identical, to an unnamed Sarcocystis sp. typified by isolates 1085 and 1086 collected from feces of a Virginia opossum (Didelphis virginiana) on the east coast of Florida. Because the life cycle of this parasite appears to be land based, urban waste discharge to marine/estuarine environments may be a source of infection for marine species.

Wading birds as bioindicators of mercury contamination in Florida, USA: annual and geographic variation.

Mercury contamination in wetland biota is often dynamic, difficult to predict, and costly to track. In this paper, we present results from a six-year study of growing feathers of piscivorous birds as monitors of wetland Hg exposure in Florida, USA, wetlands. Between 1994 and 2000, we collected feathers of growing great egret (Ardea alba) nestlings from colonies in the freshwater Everglades of southern Florida, and during 1998, feathers were collected from chicks of both great egrets and white ibises (Eudocimus albus) at a variety of colonies throughout peninsular Florida. Coastal colonies showed significantly lower feather Hg concentrations than did inland sites. Within the Everglades, we found significant effects of both geographic location and year on age-adjusted mean total Hg concentrations in feathers. Over the course of our study, Everglades colonies maintained their Hg concentration rankings relative to one another, but all showed strongly declining Hg concentrations (mean of 73% averaged across colonies, between 1994 and 2000). Using a previously established predictive relationship between Hg consumption in food and feather Hg for great egrets, we estimated that Hg concentrations in the aggregate diet of egrets have been reduced by an average of 67%. We conclude that the Everglades has undergone a biologically significant decline in Hg availability in the wetland food web, possibly because of decreased local inputs.

Burrowing fly larvae (Philornis porteri) associated with mortality of eastern bluebirds in Florida.

We investigated mortality among nestling eastern bluebirds (Sialia sialis) in Polk and Highlands counties, Florida (USA) in 1999-2001. At least six species of maggots from three families of muscoid flies, Calliphoridae, Sarcophagidae, and Muscidae were found associated with the nestlings. Philornis porteri, the only species of obligate bird parasite collected, was found in the contents of two nests, in the ear canal and the musculature of the jaw of one nestling, and in the abdominal subcutis of another. This is the first record of bluebird parasitism by P. porteri. Although some nestlings were infested by tissue-invading fly larvae antemortem, the role of these maggots in the overall mortality was not clear.