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In ovo exposure to brominated flame retardants Part II: Assessment of effects of TBBPA-BDBPE and BTBPE on hatching success, morphometric and physiological endpoints in American kestrels

Tetrabromobisphenol A bis(2,3-dibromopropyl ether) (TBBPA-BDBPE) and 1,2-bis(2,4,6-tribromophenoxy)ethane (BTPBE) are both brominated flame retardants (BFRs) that have been detected in birds; however, their potential biological effects are largely unknown. We assessed the effects of embryonic exposure to TBBPA-BDBPE and BTBPE in a model avian predator, the American kestrel ( Falco sparverius ). Fertile eggs from a captive population of kestrels were injected on embryonic day 5 (ED5) with a vehicle control or one of three doses within the range of concentrations that have been detected in biota (nominal concentrations of 0, 10, 50 or 100 ng/g egg; measured concentrations 0, 3.0, 13.7 or 33.5 ng TBBPA-BDBPE/g egg and 0, 5.3, 26.8 or 58.1 ng BTBPE/g egg). Eggs were artificially incubated until hatching (ED28), at which point blood and tissues were collected to measure morphological and physiological endpoints, including organ somatic indices, circulating and glandular thyroid hormone concentrations, thyroid gland histology, hepatic deiodinase activity, and markers of oxidative stress. Neither compound had any effects on embryo survival through 90% of the incubation period or on hatching success, body mass, organ size, or oxidative stress of hatchlings. There was evidence of sex-specific effects in the thyroid system responses to the BTBPE exposures, with type 2 deiodinase (D2) activity decreasing at higher doses in female, but not in male hatchlings, suggesting that females may be more sensitive to BTBPE. However, there were no effects of TBBPA-BDBPE on the thyroid system in kestrels. For the BTPBE study, a subset of high-dose eggs was collected throughout the incubation period to measure changes in BTBPE concentrations. There was no decrease in BTBPE over the incubation period, suggesting that BTBPE is slowly metabolized by kestrel embryos throughout their ∼28-d development. These two compounds, therefore, do not appear to be particularly toxic to embryos of the American kestrel.

Ecotoxicology and Environmental Safety↗

Toxicity of anticoagulant rodenticides on Pacific salmon: Assessing lethal and sublethal effects

To restore native biodiversity on island ecosystems containing invasive rodents, partial- and whole-island eradications generally rely on broadcast baiting with anticoagulant rodenticides (ARs). This approach can result in bait pellets entering aquatic environments, raising concerns about effects to non-target fish. Salmonids are a dominant group of fishes on many temperate islands targeted for rodent eradication, and AR toxicity data for salmonids are limited. Our goal was to determine if coho salmon ( Oncorhynchus kisutch ) are susceptible to coagulopathy and death via exposure to commonly used ARs. We assessed risk of ARs to coho using dose-response curves generated through intraperitoneal injections after determining that coho would not directly ingest the AR baits. Median lethal doses (96-h LD 50 ) estimated using 100 % corn oil carrier were 85.7 µg/g for brodifacoum and 54.0 µg/g for diphacinone. Acetone (30–41 %), used to dissolve ARs in corn oil, reduced the toxicity of diphacinone (LD 50 = 102.3 µg/g, p < 0.001) but not brodifacoum (LD 50 = 73.3 µg/g, p = 0.126) indicating that solvent choice can influence toxicity outcomes. Behavioral changes and onset of mortality differed between the two ARs, with diphacinone acting more rapidly. Tissue analysis supported a difference in toxicokinetics between the two ARs, with significant decreases in liver and muscle residues for diphacinone but not brodifacoum. Sublethal brodifacoum exposure (53.9 µg/g; LD 13 ) impaired blood clotting at 72- and 96- h but returned to baseline by 120 h. No clotting impairment was observed up to 144 h after diphacinone exposure (45.5 µg/g; LD 4 ), suggesting a non-coagulopathy mode of action. These findings will inform risk assessments when considering use of these ARs for rodent management near streams and shorelines and clearly demonstrate that brodifacoum causes coagulopathy in coho.

Ecotoxciology and Environmental Safety↗

Trace element contamination in feather and tissue samples from Anna’s hummingbirds

Trace element contamination (17 elements; Be, V, Cr, Mn, Fe, Co, Ni, Cu, Zn, As, Se, Mo, Cd, Ba, Hg, Tl, and Pb) of live (feather samples only) and deceased (feather and tissue samples) Anna's hummingbirds ( Calypte anna ) was evaluated. Samples were analyzed using inductively coupled plasma-mass spectrometry (ICP-MS; 17 elements) and atomic absorption spectrophotometry (Hg only). Mean plus one standard deviation (SD) was considered the benchmark, and concentrations above the mean + 1 SD were considered elevated above normal. Contour feathers were sampled from live birds of varying age, sex, and California locations. In order to reduce thermal impacts, minimal feathers were taken from live birds, therefore a novel method was developed for preparation of low mass feather samples for ICP-MS analysis. The study found that the novel feather preparation method enabled small mass feather samples to be analyzed for trace elements using ICP-MS. For feather samples from live birds, all trace elements, with the exception of beryllium, had concentrations above the mean + 1 SD. Important risk factors for elevated trace element concentrations in feathers of live birds were age for iron, zinc, and arsenic, and location for iron, manganese, zinc, and selenium. For samples from deceased birds, ICP-MS results from body and tail feathers were correlated for Fe, Zn, and Pb, and feather concentrations were correlated with renal (Fe, Zn, Pb) or hepatic (Hg) tissue concentrations. Results for AA spectrophotometry analyzed samples from deceased birds further supported the ICP-MS findings where a strong correlation between mercury concentrations in feather and tissue (pectoral muscle) samples was found. These study results support that sampling feathers from live free-ranging hummingbirds might be a useful, non-lethal sampling method for evaluating trace element exposure and provides a sampling alternative since their small body size limits traditional sampling of blood and tissues. The results from this study provide a benchmark for the distribution of trace element concentrations in feather and tissue samples from hummingbirds and suggests a reference mark for exceeding normal. Lastly, pollinating avian species are minimally represented in the literature as bioindicators for environmental trace element contamination. Given that trace elements can move through food chains by a variety of routes, our study indicates that hummingbirds are possible bioindicators of environmental trace element contamination.

Ecological Indicators↗

Assessment of sea lamprey (Petromyzon marinus) diet using DNA metabarcoding of feces

Sea lamprey ( Petromyzon marinus ) are invasive in the Laurentian Great Lakes, parasitize large-bodied fishes, and therefore are the focus of an international control program. However, damage caused by sea lamprey to modern day fish stocks remains uncertain because diet analysis of juvenile sea lamprey has been challenging; they feed on blood and are difficult to randomly sample in the lakes. Here, both challenges were addressed by showing that DNA metabarcoding of fecal material can be used to identify the diet of actively feeding juvenile sea lamprey, and can also be used to determine what non-feeding adult sea lamprey captured in streams fed on while parasitizing fish. Fecal samples from juvenile sea lamprey that were feeding on lake trout in northern Lake Huron overwhelmingly contained lake trout ( Salvelinus namaycush ) DNA (90%), while smaller percentages contained lake whitefish ( Coregonus clupeaformis ; 5%) and longnose sucker ( Catostomus catostomus ; 5%) DNA. Fecal samples from adult sea lamprey captured from a tributary to northern Lake Huron overwhelmingly contained longnose and white sucker DNA ( Catostomus spp.; 80%), while a smaller percentage contained lake trout DNA (10%). Diet composition of adult sea lamprey sampled in the tributary (Black Mallard Creek) was more diverse than juvenile diet composition. DNA metabarcoding suggests that Catostomus spp. may be an important host fish in northern Lake Huron for sea lamprey prior to spawning. Future research could investigate how diet varies across years and lakes and the prevalence and sources of DNA contamination. Application of DNA metabarcoding for diet assessment may be practical for identifying populations of invasive sea lamprey that feed on highly valued fishes and help guide restoration of lampreys worldwide.

Michigan↗

Establishment of baseline cytology metrics in nestling American kestrels (Falco sparverius): Immunomodulatory effects of the flame retardant isopropylated triarylphosphate isomers

Avian populations must mount effective immune responses upon exposure to environmental stressors such as avian influenza and xenobiotics. Although multiple immune assays have been tested and applied to various avian species, antibody-mediated immune responses in non-model avian species are not commonly reported due to the lack of commercially available species-specific antibodies. The objectives of the present study were to advance methods for studying wild bird immune responses and to apply these to the evaluation of cytological responses after exposure of American kestrels, Falco sparverius, to a commercial flame retardant mixture containing isopropylated triarylphosphate isomers (ITP). Hatchlings were gavaged daily with safflower oil or 1.5 ug/g bw/day of ITP suspended in safflower oil, then bled on days 9, 17, and 21. The ITP treatment group ( n = 18) and a subset of controls (Poly I:C treatment group; n = 10) were injected on days 9 and 15 with a synthetic analog of viral double-stranded RNA, polyinosinic:polycytidylic acid (Poly I:C), a toll-like receptor ligand and synthetic viral mimic, and responses compared to a sham injected control group (n = 8). The hypotheses tested whether kestrels showed immunological differences among treatment groups, genetic sex, and/or white blood cell (WBC) subpopulation type over time. A flow cytometry (FCM) gating strategy categorized heterophils (H), lymphocytes (L), and monocytes (M) and their proportions, and measured relative fluorescence in response to anti-chicken CD4 binding. Fluorescent cell surfaces and some granular/vacuolar inclusions were visualized by epifluorescence microscopy. A fourth subpopulation with higher levels of granularity than M but less than H became increasingly apparent with time and was gated along with the H subpopulation; its frequency of occurrence was lowest in the ITP group ( P = 0.0023). The percentages of cells differed among treatment groups, days, and sexes ( P = 0.0001). For both sexes, percentages of H and L were higher than M in control and Poly I:C. In the ITP group, L percentages were higher than H and M ( P = 0.0457), and H and L were higher than M on days 9 and 21 ( P = 0.0001). The ratios of H:L and H:WBC, indicators of robust immunity, were also higher on days 9 and 21 than on 17 ( P = 0.0079). For each sex, the highest levels of activity measured by FCM geometric means (GEO) of fluorescence (indicative of antibody binding) were observed on day 9 ( P = 0.0001 female, and P = 0.0011 male) in H over both L and M ( P < 0.0001 for each). In males, GEO of the Poly I:C group was higher than that of the ITP group ( P = 0.0374), with no difference observed among females over all days. By using a FCM algorithm for population comparisons of fluorescence to investigate binding within H, the T(x) scores indicated higher fluorescence in control and Poly I:C groups over ITP ( P = 0.0001). Unlike chickens, Gallus gallus , which express CD4 primarily on L, kestrels bound the commercial antibody primarily within the gated H subpopulation, suggesting an immunophenotypic difference between taxa, despite a ~60% identity of Falco CD4 amino acid sequences with chicken CD4. The emergent cell subset within the gated H presented dendritic-like cell (DLC) morphological and functional properties, apparently serving as an effector cell. This study adds interpretive context to ecological investigations of infection and of potential immunomodulation by emerging compounds, whereby the early innate responses are mediated by the various cell subsets serving as useful quantitative markers of immunological condition. Data showed that dietary exposure to ITP was immunosuppressive for male and female kestrels over the course of the experiment, reducing DLC frequency compared to the Poly I:C controls. Heterophils and DLC were important in facilitating innate immunological responses.

Environment International↗

Respiratory hazard assessment of combined exposure to complete gasoline exhaust and respirable volcanic ash in a multicellular human lung model at the air-liquid interface

Communities resident in urban areas located near active volcanoes can experience volcanic ash exposures during, and following, an eruption, in addition to sustained exposures to high concentrations of anthropogenic air pollutants ( e.g., vehicle exhaust emissions). Inhalation of anthropogenic pollution is known to cause the onset of, or exacerbate, respiratory and cardiovascular diseases. It is further postulated similar exposure to volcanic ash can also affect such disease states. Understanding of the impact of combined exposure of volcanic ash and anthropogenic pollution to human health, however, remains limited. The aim of this study was to assess the biological impact of combined exposure to respirable volcanic ash (from Soufrière Hills volcano (SHV), Montserrat and Chaitén volcano (ChV), Chile; representing different magmatic compositions and eruption styles) and freshly-generated complete exhaust from a gasoline vehicle. A multicellular human lung model (an epithelial cell-layer composed of A549 alveolar type II-like cells complemented with human blood monocyte-derived macrophages and dendritic cells cultured at the air-liquid interface) was exposed to diluted exhaust (1:10) continuously for 6 h, followed by immediate exposure to the ash as a dry powder (0.54 ± 0.19 μg/cm 2 and 0.39 ± 0.09 μg/cm 2 for SHV and ChV ash, respectively). After an 18 h incubation, cells were exposed again for 6 h to diluted exhaust, and a final 18 h incubation (at 37 °C and 5% CO 2 ). Cell cultures were then assessed for cytotoxic, oxidative stress and (pro-)inflammatory responses. Results indicate that, at all tested (sub-lethal) concentrations, co-exposures with both ash samples induced no significant expression of genes associated with oxidative stress ( HMOX1, NQO1 ) or production of (pro-)inflammatory markers (IL-1β, IL-8, TNF-α) at the gene and protein levels. In summary, considering the employed experimental conditions, combined exposure of volcanic ash and gasoline vehicle exhaust has a limited short-term biological impact to an advanced lung cell in vitro model.

Environmental Pollution↗

Arsenic-related oxidative stress in experimentally dosed wild great tit nestlings

Arsenic (As) is broadly distributed due to natural and anthropogenic sources, and it may cause adverse effects in birds. However, research on other elements (Pb, Hg and Cd) has been prioritized, resulting in scarce data on As exposure and related effects in wild birds. One of the mechanisms responsible for As toxicity is oxidative stress. Therefore, the aim of this study was to investigate if environmentally relevant As levels affected oxidative stress biomarkers in great tits ( Parus major ). This is the first field experiment studying the effects of As on oxidative stress in wild passerines. Wild great tit nestlings were orally dosed with sodium arsenite (Control: water, Low dose: 0.2 μg g −1 d −1 and High dose: 1 μg g −1 d −1 ; from day 3 to day 13 post-hatching). We intended to reach As concentrations similar to those at which passerines are exposed to at actual polluted areas. We compared the responses to the experimental manipulations (High, Low and Control groups) with those in an As/metal-exposed population breeding close to a Cu–Ni smelter in Finland (Smelter group). A set of antioxidants (tGSH, GSH:GSSG ratio, CAT, SOD, GST and GPx), and oxidative damage biomarkers (lipid peroxidation, protein carbonylation, 8-hydroxy-2′-deoxyguanosine formation in DNA, and telomere length) were explored in blood. Arsenic administration had no significant effect on most of the biomarkers measured: only the CAT activity was lower in the High As group and the GPx activity was enhanced in the Smelter group compared to the Control. Our results suggest that the dose and duration of the As exposure was not enough to induce oxidative damage in red cells of great tit nestlings. In spite of this, nestlings dosed with 1 μg g −1 d −1 of sodium arsenite showed non-significantly higher oxidative stress biomarkers than controls, suggesting that we were close to an effect level for the redox-defense system. Oxidative effects at equivalent As levels combined with other stressors cannot be dismissed.

Harjavalta↗

Per- and polyfluoroalkyl substances and pesticides in black bears (Ursus americanus) and grizzly bears (Ursus arctos horribilis) from Montana's Cabinet-Yaak ecosystem: A baseline assessment of emerging and legacy contaminants

Per- and polyfluoroalkyl substances (PFAS) and pesticides are globally distributed contaminants that persist in terrestrial food webs, yet baseline data for large omnivores in the continental U.S. remain limited. This study quantified PFAS and pesticides in whole blood from free-ranging black bears ( Ursus americanus , n = 15) and grizzly bears ( Ursus arctos horribilis , n = 9) in the Cabinet-Yaak Ecosystem of northwestern Montana. Using high-resolution mass spectrometry-based methods (GC-MS/MS and LC-MS/MS), 52 PFAS and 186 pesticides were analyzed. PFAS were detected in all bears, with total PFAS concentrations ranging from 79.5 to 317 pg/mL. PFAS profiles were dominated by long-chain perfluoroalkyl carboxylic acids, particularly PFOA, PFDA, and PFUnDA, with minimal contribution from short-chain compounds. Species was the strongest ecological predictor of PFAS concentrations, although variability across age, sex, and capture location were observed. Only two pesticides, piperonyl butoxide and permethrin, were detected at quantifiable concentrations (2070–12,600 pg/mL), and no correlations were observed between pesticide and PFAS concentrations, indicating independent exposure pathways. The predominance of long-chain PFAS suggests diffuse environmental sources and bioaccumulation within terrestrial food webs. Although measured concentrations were generally low relative to other wildlife toxicological studies, the persistence and protein-binding properties of long-chain PFAS warrant continued monitoring, particularly for long-lived omnivores. These results establish baseline contaminant concentrations for two apex omnivores in a remote ecosystem and highlight the utility of large mammals as integrative sentinels of environmental contaminant exposure for regional monitoring and wildlife conservation. Continued monitoring in the Cabinet-Yaak and across western North America will be critical for assessing temporal trends, identifying new contaminant sources, and evaluating ecological health in bear populations.

Montana↗

The effect of aluminium and sodium impurities on the in vitro toxicity and pro-inflammatory potential of cristobalite

Background Exposure to crystalline silica (SiO 2 ), in the form of quartz, tridymite or cristobalite, can cause respiratory diseases, such as silicosis. However, the observed toxicity and pathogenicity of crystalline silica is highly variable. This has been attributed to a number of inherent and external factors, including the presence of impurities. In cristobalite-rich dusts, substitutions of aluminium (Al) for silicon (Si) in the cristobalite structure, and impurities occluding the silica surface, have been hypothesised to decrease its toxicity. This hypothesis is tested here through the characterisation and in vitro toxicological study of synthesised cristobalite with incremental amounts of Al and sodium (Na) dopants. Methods Samples of synthetic cristobalite with incremental amounts of Al and Na impurities, and tridymite, were produced through heating of a silica sol-gel. Samples were characterised for mineralogy, cristobalite purity and abundance, particle size, surface area and surface charge. In vitro assays assessed the ability of the samples to induce cytotoxicity and TNF-α production in J774 macrophages, and haemolysis of red blood cells. Results Al-only doped or Al+Na co-doped cristobalite contained between 1 and 4 oxide wt% Al and Na within its structure. Co-doped samples also contained Al- and Na-rich phases, such as albite. Doping reduced cytotoxicity to J774 macrophages and haemolytic capacity compared to non-doped samples. Al-only doping was more effective at decreasing cristobalite reactivity than Al+Na co-doping. The reduction in the reactivity of cristobalite is attributed to both structural impurities and a lower abundance of crystalline silica in doped samples. Neither non-doped nor doped crystalline silica induced production of the pro-inflammatory cytokine TNF-α in J774 macrophages. Conclusions Impurities can reduce the toxic potential of cristobalite and may help explain the low reactivity of some cristobalite-rich dusts. Whilst further work is required to determine if these effects translate to altered pathogenesis, the results have potential implications for the regulation of crystalline silica exposures.

Environmental Research↗

Physiological response of wild rainbow trout to angling: Impact of angling duration, fish size, body condition, and temperature

This study evaluated the immediate physiological response of wild rainbow trout to catch-and-release angling in the Alagnak River, southwest Alaska. Information was recorded on individual rainbow trout (n = 415) captured by angling including landing time and the time required to remove hooks (angling duration), the time to anesthetize fish in clove oil and withdraw blood, fish length and weight, and water temperature at capture locations. Plasma cortisol, glucose, ions (sodium, potassium, chloride), and lactate were analyzed to determine the effects of angling duration, fish size, body condition, and temperature. Levels of plasma ions did not change significantly during the observed physiological response and levels of plasma glucose were sometimes influenced by length (2000, 2001), body condition (2001), or temperature (2001). Levels of plasma cortisol and lactate in extended capture fish (angling duration greater than 2 min) were significantly higher than levels in rapid capture fish (angling duration less than 2 min). Rapid capture fish were significantly smaller than extended capture fish, reflecting that fish size influenced landing and handling times. Fish size was related to cortisol and lactate in 2002, which corresponded to the year when larger fish were captured and there were longer landing times. Body condition (i.e., weight/length regression residuals index), was significantly related to lactate in 2000 and 2001. Water temperatures were higher in 2001 (mean temperature &plusmn; S.E., 13 &plusmn; 2 o C) than in 2002 (10 &plusmn; 2 o C), and fish captured in 2001 had significantly higher cortisol and lactate concentrations than fish captured in 2002. The pattern of increase in plasma cortisol and lactate was due to the amount of time fish were angled, and the upper limit of the response was due to water temperature. The results of this study indicate the importance of minimizing the duration of angling in order to reduce the sublethal physiological disturbances in wild fish subjected to catch-and-release angling, particularly during warmer water temperatures. It is also important to note that factors such as fish size may influence both the duration of angling and subsequent physiological response.

Fisheries Research↗

Channel catfish (Ictalurus punctatus) leukocytes express estrogen receptor isoforms ERα and ERβ2 and are functionally modulated by estrogens

Estrogens are recognized as modulators of immune responses in mammals and teleosts. While it is known that the effects of estrogens are mediated via leukocyte-specific estrogen receptors (ERs) in humans and mice, leucocyte-specific estrogen receptor expression and the effects of estrogens on this cell population is less explored and poorly understood in teleosts. Here in, we verify that channel catfish ( Ictalurus punctaus ) leukocytes express ERα and ERβ2. Transcripts of these isoforms were detected in tissue-associated leukocyte populations by PCR, but ERβ2 was rarely detected in PBLs. Expression of these receptors was temporally regulated in PBLs following polyclonal activation by concanavalin A, lipopolysaccharide or alloantigen based on evaluation by quantitative and end-point PCR. Examination of long-term leukocyte cell lines demonstrated that these receptors are differentially expressed depending on leukocyte lineage and phenotype. Expression of ERs was also temporally dynamic in some leukocyte lineages and may reflect stage of cell maturity. Estrogens affect the responsiveness of channel catfish peripheral blood leukocytes (PBLs) to mitogens in vitro . Similarly, bactericidal activity and phorbol 12-myristate 13-acetate induced respiratory burst was modulated by 17β-estradiol. These actions were blocked by the pure ER antagonist ICI 182780 indicating that response is, in part, mediated via ERα. In summary, estrogen receptors are expressed in channel catfish leukocytes and participate in the regulation of the immune response. This is the first time leukocyte lineage expression has been reported in teleost cell lines.

Fish & Shellfish Immunology↗

Avian malaria in a boreal resident species: long-term temporal variability, and increased prevalence in birds with avian keratin disorder

The prevalence of vector-borne parasitic diseases is widely influenced by biological and ecological factors. Environmental conditions such as temperature and precipitation can have a marked effect on haemosporidian parasites ( Plasmodium spp.) that cause malaria and those that cause other malaria-like diseases in birds. However, there have been few long-term studies monitoring haemosporidian infections in birds in northern latitudes, where weather conditions can be highly variable and the effects of climate change are becoming more pronounced. We used molecular methods to screen more than 2,000 blood samples collected from black-capped chickadees ( Poecile atricapillus ), a resident passerine bird. Samples were collected over a 10 year period, mostly during the non-breeding season, at seven sites in Alaska, USA. We tested for associations between Plasmodium prevalence and local environmental conditions including temperature, precipitation, site, year and season. We also evaluated the relationship between parasite prevalence and individual host factors of age, sex and presence or absence of avian keratin disorder. This disease, which causes accelerated keratin growth in the beak, provided a natural study system in which to test the interaction between disease state and malaria prevalence. Prevalence of Plasmodium infection varied by year, site, age and individual disease status but there was no support for an effect of sex or seasonal period. Significantly, birds with avian keratin disorder were 2.6 times more likely to be infected by Plasmodium than birds without the disorder. Interannual variation in the prevalence of Plasmodium infection at different sites was positively correlated with summer temperatures at the local but not statewide scale. Sequence analysis of the parasite cytochrome b gene revealed a single Plasmodium spp. lineage, P43. Our results demonstrate associations between prevalence of avian malaria and a variety of biological and ecological factors. These results also provide important baseline data that will be informative for predicting future changes in Plasmodium prevalence in the subarctic.

Alaska↗

Enhanced understanding of ectoparasite: host trophic linkages on coral reefs through stable isotope analysis

Parasitism, although the most common type of ecological interaction, is usually ignored in food web models and studies of trophic connectivity. Stable isotope analysis is widely used in assessing the flow of energy in ecological communities and thus is a potentially valuable tool in understanding the cryptic trophic relationships mediated by parasites. In an effort to assess the utility of stable isotope analysis in understanding the role of parasites in complex coral-reef trophic systems, we performed stable isotope analysis on three common Caribbean reef fish hosts and two kinds of ectoparasitic isopods: temporarily parasitic gnathiids ( Gnathia marleyi ) and permanently parasitic cymothoids ( Anilocra ) . To further track the transfer of fish-derived carbon (energy) from parasites to parasite consumers, gnathiids from host fish were also fed to captive Pederson shrimp ( Ancylomenes pedersoni ) for at least 1 month. Parasitic isopods had &delta; 13 C and &delta; 15 N values similar to their host, comparable with results from the small number of other host&ndash;parasite studies that have employed stable isotopes. Adult gnathiids were enriched in 15 N and depleted in 13 C relative to juvenile gnathiids, providing insights into the potential isotopic fractionation associated with blood-meal assimilation and subsequent metamorphosis. Gnathiid-fed Pedersen shrimp also had &delta; 13 C values consistent with their food source and enriched in 15 N as predicted due to trophic fractionation. These results further indicate that stable isotopes can be an effective tool in deciphering cryptic feeding relationships involving parasites and their consumers, and the role of parasites and cleaners in carbon transfer in coral-reef ecosystems specifically.

Lameshur Bay, St. John, Virgin Islands↗

Negligible evidence for detrimental effects of Leucocytozoon infections among Emperor Geese (Anser canagicus) breeding on the Yukon-Kuskokwim Delta, Alaska

Emperor Geese ( Anser canagicus ) are iconic waterfowl endemic to Alaska and adjacent areas of northeastern Russia that are considered to be near threatened by the International Union for Conservation. This species has been identified as harboring diverse viruses and parasites which have, at times, been associated with disease in other avian taxa. To better assess if disease represents a vulnerability for Emperor Geese breeding on the Yukon-Kuskokwim Delta, Alaska, we evaluated if haemosporidian parasites were associated with decreased mass or survival among adult female nesting birds captured during 2006–2016. Through molecular analyses, we detected genetically diverse Leucocytozoon , Haemoproteus , and Plasmodium parasites in 28%, 1%, and 1% of 607 blood samples screened in triplicate, respectively. Using regression analysis, we found evidence for a small effect of Leucocytozoon infection on the mass of incubating adult female Emperor Geese. The estimated mass of infected individuals was approximately 43 g (95% CI: 20–67 g), or approximately 2%, less than uninfected birds when captured during the second half of incubation (days 11–25). We did not, however, find support for an effect of Leucocytozoon infection on survival of adult female nesting Emperor Geese using a multi-state hidden Markov framework to analyze mark-resight and recapture data. Using parasite mitochondrial DNA cytochrome b sequences, we identified 23 haplotypes among infected Emperor Geese. Leucocytozoon haplotypes clustered into three phylogenetically supported clades designated as ‘ L. simondi clade A’, ‘ L. simondi clade B’, and ‘other Leucocytozoon ’. We did not find evidence that parasites assigned to any of these clades were associated with differential mass measures among nesting adult female Emperor Geese. Collectively, our results provide negligible evidence for Leucocytozoon parasites as causing detrimental effects to adult female Emperor Geese breeding on the Yukon-Kuskokwim Delta.

Alaska↗

Contrasting haemosporidian infections in two ecologically distinct wading birds from breeding colonies in the southeastern United States

Wading birds may serve as ideal hosts for avian hemoparasites, as they are long-lived, undertake extensive movements, form dense breeding colonies, and inhabit water-associated environments that support vectors. Although previous studies have reported parasite species and prevalence in various wading bird species, little is known about their associations with bird behavior and life stage. To address this gap, we examined haemosporidian infections in two ecologically distinct species, white ibis ( Eudocimus albus ) and tricolored heron ( Egretta tricolor ), to explore differences in life stage and movement that may explain prevalence differences. We combined blood screening for hemoparasites with satellite tracking data describing birds’ movement patterns. We screened 95 white ibis (67 juveniles and 28 adults or subadults) and 69 tricolored herons (45 juveniles and 24 adults). We detected a single Haemoproteus plataleae lineage in both species, with higher infection prevalence in white ibis (42.1 %) than in tricolored herons (14.5 %). Among white ibis, adults showed a higher prevalence (67.9 %) than juveniles (31.3 %), whereas in tricolored herons, adults had a prevalence of 8.3 % and juveniles 17.8 %. Non-breeding season movement data showed that white ibis used both freshwater and saline habitats across the southeastern United States, which may also serve as habitats for vectors. In contrast, tricolored herons remained mainly along coastal areas, using saline habitats in the southeastern United States and wintering sites in Central America, which may be less favorable for vectors. Overall, white ibis may serve as major reservoirs and sources of reinfection for H. plataleae , whereas tricolored herons may facilitate parasite dispersal between breeding colonies along the coasts of the southeastern United States and wintering areas in Central America. This study presents the first direct comparison of hemoparasite infections in two ecologically distinct wading birds and highlights movement data as key to explaining infection differences, providing a baseline for future studies.

Alabama↗

Serum iron analytes in healthy and diseased Florida manatees (Trichechus manatus latirostris)

Serum iron concentration is usually decreased in true iron deficiency and with inflammatory disease in man and domestic animals. Serum total iron binding capacity (TIBC) may be increased in true iron deficiency and decreased with inflammatory disease. This prospective study was designed to measure serum iron analytes in healthy free-ranging and housed Florida manatees (Trichechus manatus latirostris) of both sexes and various ages and to evaluate the effects of diseases common to manatees on these analytes. Blood samples were collected without anticoagulant from 137 healthy free-ranging manatees, 90 healthy housed manatees and 74 free-ranging diseased manatees, and serum was prepared by centrifugation. Serum iron concentration and unsaturated iron binding capacity were measured colourimetrically, and TIBC and percent transferrin saturation with iron were calculated. Serum amyloid A (SAA) was measured to assist in the health assessment of manatees and provide evidence of inflammation in diseased manatees. Based on the serum iron analytes, iron availability was lower in immature manatees compared with adults, and it was lower in housed manatees compared with free-ranging manatees. In contrast to other mammals studied, serum iron concentration was elevated rather than depressed in late pregnancy. Serum iron concentrations and transferrin saturation with iron percentages were significantly lower, and SAA concentrations were significantly higher, in diseased (ill and injured) manatees compared with healthy manatees. Serum iron concentration and transferrin saturation with iron values were negatively correlated with SAA concentrations, and manatees with the highest SAA concentrations had lower serum TIBC values. These findings indicate that inflammation is the major factor responsible for alterations in iron analytes in diseased manatees. Consequently, hypoferraemia may be used as supportive evidence of inflammatory disease in manatees (unless haemorrhage is also present). A decision threshold of ≤13.8 μmol/l was determined for hypoferraemia using receiver operating curve analysis. Based on studies in man and domestic animals, iron therapy is unnecessary for manatees with hypoferraemia associated with inflammation and has the potential for causing tissue damage and increased susceptibility to bacterial infections.

Florida↗

Evidence of chromosomal damage in common eiders (Somateria mollissima) from the Baltic Sea

Common eiders nesting in the Baltic Sea are exposed to generally high levels of contaminants including potentially genotoxic polycyclic aromatic hydrocarbons and organochlorines. Blood samples were collected from eiders at eight sites in the Baltic Sea and two sites in the Beaufort Sea. DNA content variation was estimated using the flow cytometric method, and subsequently utilized as a biomarker of genetic damage. We observed no significant differences in genetic damage among populations within either the Baltic or Beaufort Seas. However, eider populations from the Baltic Sea had significantly elevated estimates of genetic damage compared to populations from the Beaufort Sea.

Baltic Sea, Beaufort Seas↗

Selenium and mercury concentrations in harbor seals (Phoca vitulina) from central California: Health implications in an urbanized estuary

We measured total selenium and total mercury concentrations ([TSe] and [THg]) in hair (n = 138) and blood (n = 73) of harbor seals ( Phoca vitulina ) from California to assess variation by geography and sex, and inferred feeding relationships based on carbon, nitrogen, and sulfur stable isotopes. Harbor seals from Hg-contaminated sites had significantly greater [THg], and lesser [TSe] and TSe:THg molar ratios than seals from a relatively uncontaminated site. Males had significantly greater [THg] than females at all locations. Sulfur stable isotope values explained approximately 25% of the variability in [THg], indicating increased Hg exposure for seals with a greater use of estuarine prey species. Decreased [TSe] in harbor seals from Hg-contaminated regions may indicate a relative Se deficiency to mitigate the toxic effects of Hg. Further investigation into the Se status and the potential negative impact of Hg on harbor seals from Hg-contaminated sites is warranted.

California↗