Copper (Cu) is a vital micronutrient required for numerous fundamental biological processes, but excessive Cu poses potential detrimental effects on public and ecosystem health. However, the molecular details linking endoplasmic reticulum (ER) stress and apoptosis in duck renal tubular epithelial cells have not been fully elucidated. In this study, duck renal tubular epithelial cells exposed to Cu sulfate (CuSO₄) (0, 100 and 200 μM) and a PERK inhibitor (GSK2606414, GSK, 1 μM) for 12 h were used to investigate the crosstalk between ER stress and apoptosis under Cu exposure. Cell and ER morphological and functional characteristics, intracellular calcium (Ca²⁺) levels, apoptotic rates, ER stress and apoptosis-related mRNA and protein levels were examined. The results showed that excessive Cu could cause ER expansion and swelling, increase the expression levels of ER stress-associated genes (PERK, eIF2α, ATF4 and CHOP) and proteins (p-PERK and CHOP), induce intracellular Ca²⁺ overload, upregulate the expression levels of apoptosis-associated genes (Bax, Bak1, Caspase9 and Caspase3) and the cleaved-Caspase3 protein, downregulate Bcl-xl and Bcl2 mRNA levels and trigger apoptosis. PERK inhibitor treatment could ameliorate the above changed factors caused by Cu. In conclusion, these findings indicate that excessive Cu could trigger ER stress via activation of the PERK/ATF4/CHOP signaling pathway and that ER stress might aggravate Cu-induced apoptosis in duck renal tubular epithelial cells.

In this paper we present the studies conducted on blood samples taken from Mallards (Anas platyrhynchos). Birds were captured for ringing purposes (n = 43) in two small and two big towns (including highly urbanized areas). For comparison samples of blood from birds shot on fish ponds were used (n = 26). Based on the body mass all sampled individuals can be assessed as being in good condition. Levels of cadmium (Cd), chromium (Cr), copper (Cu), iron (Fe), nickel (Ni), lead (Pb) and zinc (Zn) in blood samples were measured with AAS. Concentrations of metals did not differ statistically between sexes and made up a following order: Fe > Zn > Cu > Cr ≈ Ni > Pb > Cd. Mallards from towns revealed lower concentrations of Zn and Cu but higher concentration of Fe. There was no difference in exposition to Pb between birds from towns and fish ponds.

In this study, total mercury (THg) and methylmercury (MeHg) concentrations in muscles (leg and breast), organs (intestine, heart, stomach, liver) and blood were investigated for backyard chickens, ducks and geese of the Wanshan Mercury Mine, China. THg in poultry meat products range from 7.9 to 3917.1 ng/g, most of which exceeded the Chinese national standard limit for THg in meat (50 ng/g). Elevated MeHg concentrations (0.4–62.8 ng/g) were also observed in meat products, suggesting that poultry meat can be an important human MeHg exposure source. Ducks and geese showed higher Hg levels than chickens. For all poultry species, the highest Hg concentrations were observed in liver (THg: 23.2–3917.1 ng/g; MeHg: 7.1–62.8 ng/g) and blood (THg: 12.3–338.0 ng/g; MeHg: 1.4–17.6 ng/g). We estimated the Hg burdens in chickens (THg: 15.3–238.1 μg; MeHg: 2.2–15.6 μg), ducks (THg: 15.3–238.1 μg; MeHg: 3.5–14.7 μg) and geese (THg: 83.8–93.4 μg; MeHg: 15.4–29.7 μg). To not exceed the daily intake limit for THg (34.2 μg/day) and MeHg (6 μg/day), we suggested that the maximum amount (g) for chicken leg, breast, heart, stomach, intestine, liver, and blood should be 1384, 1498, 2315, 1214, 1081, 257, and 717, respectively; the maximum amount (g) for duck leg, breast, heart, stomach, intestine, liver, and blood should be 750, 1041, 986, 858, 752, 134, and 573, respectively; and the maximum amount (g) for goose leg, breast, heart, stomach, intestine, liver, and blood should be 941, 1051, 1040, 1131, 964, 137, and 562, respectively.

This study aimed to investigate the accumulation of polybrominated diphenyl ethers (PBDEs) in the brain compared with that in other tissues among different vertebrates. We collected mice, chickens, ducks, frogs, and fish from an e-waste recycling region in Taizhou, China, and measured PBDE concentrations in brain, liver and muscle tissues. The levels of PBDE in the tissues of mice, chickens, ducks, frogs and fish ranged 0.45–206, 0.06–18.8, 1.83–112, 2.75–108, and 0.02–32.0 ng/g wet weight, respectively. Preferential distribution in the liver and muscle relative to the brain was observed for PBDEs in mice, chickens, ducks and frogs. However, a high retention in the brain compared to the liver and muscle was observed in fish. Comparison of the brain/liver concentration (B/L) ratios revealed differences in PBDEs accumulation in the brain among these vertebrates. PBDEs accumulation in the brain was greatest in fish, followed by frogs, while the lowest accumulation occurred in the brains of mammals and birds. The findings apparently coincided with the evolution of the blood-brain barrier (BBB) across vertebrates, i.e. the BBB of fish might be less efficient than those of mammals, birds and amphibian. Low brominated congeners (such as BDE-28, BDE-47 and BDE-99) were predominant in the brains of investigated vertebrates, whereas BDE-209 was most abundant in liver and muscle tissues of mice, chickens and ducks. Significant differences in B/L ratios among PBDE congeners were found in both mice and chickens (p < 0.05). Particularly in mice, the B/L ratios of PBDE congeners presented a declining trend with increased bromine number. Our findings suggested that low brominated congeners might have a higher capacity to penetrate the BBB and accumulate in the brain, whereas high brominated congeners such as BDE-209 might have less potency to pass through the barrier. Further experimental studies are needed to confirm our findings.

The Hudson River, NY, USA is contaminated for over 300 km with polychlorinated biphenyls (PCBs) released from two General Electric (GE) capacitor plants. We collected adult and juvenile mallards (Anas platyrhynchos) from four different areas of the river; an area upstream of the GE plants (n = 38), two areas directly downstream of the GE plants (n = 41, n = 38), and an area more than 100 km downstream in the freshwater tidal river (n = 20). Collections occurred during July and August (2008) when ducks were flightless to ensure ducks were “resident” and exposures were local. Fat and muscle tissue were analyzed for PCBs. PCBs were detected in all samples, and mallards below the GE plant sites on the Hudson River had orders of magnitude higher concentrations of PCBs than those above the plants. Juvenile mallards from areas directly downstream of the GE plant sites tended to have higher PCB concentrations in fat than adults. The patterns of PCB congeners and homolog groups varied across the study areas, with areas directly downstream of the GE plants dominated by tetra-chloro biphenyls whereas samples from upstream and the freshwater tidal river tended towards higher chlorinated congeners. Congener patterns between male and female and juvenile and adult mallards were generally similar within study areas, with the exception of one area downstream of the GE plants where adult birds exhibited different patterns than juveniles. Evidence of PCBs from the GE plant sites was detected in the tidal Hudson River, more than 100 km downstream of the plant sites. More than 90% of the ducks collected in areas downstream of the GE plants but above the tidally influenced river exceed the USFDA tolerance level for PCBs in poultry, which should be a concern for consumers of waterfowl taken in proximity to the upper Hudson River.

Accumulation of selenium (Se) by lesser and greater scaup (Aythya affinis, A. marila) at staging and wintering areas could have contributed to the decline in their continental population. We exposed lesser scaup to background (0.8 μg/g), moderate (8.1 μg/g) and high (20.7 μg/g) levels of dietary Se in captivity and measured survival rates and indices of health in relation to hepatic Se concentrations. There was 100% survival in scaup exposed to Se for 10-weeks (average staging duration at Great Lakes), but ducks in the high treatment group had less lipids. There was 93% survival after 23-weeks (average wintering duration at Great Lakes), but no differences among treatment groups in body composition. There were no effects of Se on oxidative stress and cell-mediated immunity; rather we recorded immuno-stimulatory effects on antibody production. Results from our captive study suggest Se alone did not cause the continental decline in scaup populations.

Elevated levels of selenium (Se) have been detected in wintering and spring-staging lesser scaup. Here, we compared spring scaup Se and mercury (Hg) levels to those of ring-necked ducks and white-winged scoters, species exhibiting increasing and decreasing boreal populations, respectively. Mercury concentrations were low in all three species. Geometric mean (95%CI) liver Se concentrations were 6.2 (5.5-7.0), 4.6 (4.0-5.4), and 32.6 (28.4-37.3) mg/kg dry weight (dw) in scaup, ringnecks and scoters, respectively. Only scoter livers (66%) were above 33 mg/kg dw Se. Scaup and ringneck Se levels were unrelated to breeding status or lipid and protein levels; breeding scoters and females with greater lipid mass had higher Se than non-breeders. Egg and follicle concentrations in scaup and scoters were normal (mean [95%CI] = 2.3 [1.9-2.6] and 2.4 [2.1-2.7] mg/kg dw, respectively). Overall, we found no support for a relationship between selenium and boreal scaup and scoter declines, and discuss current Se threshold concentrations. Selenium is not affecting boreal scaup, ring-necked duck, or scoter body condition or reproduction.

Among wild birds, lead (Pb) exposure caused by ingestion of ammunition is a worldwide problem. We aimed to reveal the behavior and toxic effect of Pb caused by ingesting Pb shots in waterfowl. Four male, eight-week old Muscovy ducks (Cairina moschata) were given three Pb shots (approximately 240 mg in total) orally and then fed for 29 days after exposure, simulating a low-dose Pb exposure in wild waterfowl. During the breeding period, blood samples were collected 10 times, and fecal samples every day. Additionally, 22 fresh tissue and 6 bone samples were obtained from each duck through the dissection. Although there were no gross abnormalities, the maximum blood Pb concentration of each duck ranged from 0.6 to 3.7 mg/L, reaching a threshold concentration indicative of clinical symptoms (>0.5 mg/L). δ-aminolevulinic acid dehydratase declined one day after exposure and remained low throughout the feeding period. Hematocrit also tended to decrease, indicating signs of anemia. The highest Pb accumulation was observed in the bones, followed by the kidneys, intestinal tracts, and liver. High Pb accumulation in the bones, which are known to have a long Pb half-life, suggested that Pb would remain in the body and possibly affect bird health beyond 28 days after exposure. Gene expression analysis showed a significant increase in the expression of the toll-like receptor-3 gene, which is involved in virus discrimination in the liver, suggesting a disruption of the immune system. Microbiota analyses showed a correlation between the blood Pb concentration and the abundances of Lachnospiraceae and Ruminococcaceae, suggesting that Pb affects lipid metabolism. These results provide fundamental data on Pb exposure in wild birds and a new perspective on the damage such exposure causes.

Cadmium (Cd) and excess molybdenum (Mo) are harmful to animals, but the combined nephrotoxic mechanism of Cd and Mo in duck remains poorly elucidated. To assess joint effects of Cd and Mo on pyroptosis via ROS/PTEN/PI3K/AKT axis in duck renal tubular epithelial cells, cells were cultured with 3CdSO₄·8H₂O (4.0 μM), (NH₄)₆Mo₇O₂₄·4H₂O (500.0 μM), MCC950 (10.0 μM), BHA (100.0 μM) and combination of Cd and Mo or Cd, Mo and MCC950 or Cd, Mo and BHA for 12 h, and the joint cytotoxicity was explored. The results manifested that toxicity of non-equitoxic binary mixtures of Mo and Cd exhibited synergic interaction. Mo or/and Cd elevated ROS level, PTEN mRNA and protein levels, and decreased PI3K, AKT and p-AKT expression levels. Simultaneously, Mo or/and Cd upregulated ASC, NLRP3, NEK7, Caspase-1, GSDMA, GSDME, IL-18 and IL-1β mRNA levels and Caspase-1 p20, NLRP3, ASC, GSDMD protein levels, increased the percentage of pyroptotic cells, LDH, NO, IL-18 and IL-1β releases as well as relative conductivity. Moreover, NLRP3 inhibitor MCC950 and ROS scavenger BHA could ameliorate the above changed factors induced by Mo and Cd co-exposure. Collectively, our results reveal that combination of Mo and Cd synergistically cause oxidative stress and trigger pyroptosis via ROS/PTEN/PI3K/AKT axis in duck tubular epithelial cells.