Sub-lethal impacts of lead poisoning on blood biochemistry, immune function and delta-aminolevulinic acid dehydratase (δ-ALAD) activity in Cape (Gyps coprotheres) and white-backed (G. africanus) Vulture chicks.

Although the prevalence of lead poisoning in southern Africa's Gyps vultures is now well-established, its finer physiological effects on these endangered species remain poorly characterised. We evaluated the sub-lethal impact of acute lead exposure on Cape and White-backed Vulture chicks from two breeding colonies in South Africa, by analysing its possible effects on key blood biochemistry parameters, immune function, packed cell volume and δ-aminolevulinic acid dehydratase (δ-ALAD) activity. All 37 White-backed Vulture nestlings sampled displayed elevated lead levels (>10 µg/dL), and seven had blood [Pb] >100 µg/dL. Eight of 28 Cape Vulture nestlings sampled had blood [Pb] exceeding background exposure, with one showing blood [Pb] >100 µg/dL. Delta-aminolevulinic acid dehydratase (δ-ALAD) activity was significantly and negatively related to blood [Pb] in nestlings from both species, with 50% inhibition of the enzyme predicted to occur at blood [Pb] = 52.8 µg/dL (White-backed Vulture) and 18.8 µg/dL (Cape Vulture). Although no significant relationship was found between % packed cell volume (PCV) and blood [Pb], the relatively lower mean PCV of 32.9% in White-backed Vulture chicks, combined with normal serum protein values, is likely indicative of depression or haemolytic anaemia. The leukogram was consistent in both species, although the presence of immature heterophils suggested an inflammatory response in White-backed Vulture chicks with blood [Pb] >100 µg/dL. Values for cholesterol, triglycerides, total serum protein, albumin, globulin, albumin/globulin ratio, alanine aminotransferase (ALT) and gamma-glutamyl transferase (GGT) were consistent with values previously reported. Calcium and phosphorus concentrations suggested no adverse effects on bone metabolism. A significant decrease in urea: uric acid (U:UA) ratio at blood [Pb] >100 µg/dL in White-backed Vulture chicks, brought about by a decrease in urea production, raises the possibility of hepatic abnormality. These results suggest that δ-ALAD activity may serve as a sensitive biomarker of lead toxicity in both species, while highlighting the need to better understand the significant variability in sensitivity that is observed, even between closely related members of the same genus.

Using blood lead concentrations of wildlife sentinels to identify environmental risk factors of lead exposure for public health and wildlife rehabilitation efforts.

Lead poisoning remains a persistent health issue for both humans and wildlife, despite strides to reduce lead contamination in the environment. Using Geographic Information Systems (GIS), this study explores the associations between blood lead levels (BLLs) in wildlife sentinels and possible built environment lead exposure risk factors in the Minneapolis-Saint Paul, Minnesota urban area. Results show a high-level of heterogeneity in animal BLLs (n = 472) across our urban environment and suggest that each kilometer increase in road density is associated with a 17.07% (95% CI: 1.48%, 35.05%) increase in BLL in our study species of Virginia opossums and Eastern gray squirrels, and a 14.28% (95% CI: 1.16%, 29.09%) increase in BLL of rock pigeons. For squirrels and opossums, we see an additional 5.72% (95% CI: 0.59%, 10.85%) increased risk of BLL for every 1000 people per square-mile. The relationship between animal sentinels and environmental hazards can give us an insight into the potential lead exposure risks for humans. The use of wildlife sentinel data to explore environmental risk factors supports a One Health approach to better address public health questions and aid in wildlife rehabilitation related to residual lead poisoning from ambient environmental exposures.

Expanding the use of portable XRF to monitor lead exposure in an Australian duck species two decades after a ban on lead shot.

There is growing worldwide recognition of the threat posed by toxic lead for wildlife and humans. Lead toxicity from ammunition has been shown to be a threat to waterbirds across the globe. Lead shot was banned for all waterfowl hunting in Victoria, Australia, in 2002. However, no assessments of lead exposure in Australian waterfowl have been published since the 1990s. Our aim was to estimate contemporary lead exposure via measuring bone lead concentrations in a harvested dabbling duck, the Pacific black duck (Anas superciliosa). We collected wings from 77 Pacific black ducks, spanning 2018 (n = 30) and 2021 (n = 47), from nine sites with long-term histories of regular waterfowl hunting. We sought to validate portable X-ray fluorescence (XRF) for this purpose by taking a piece of humerus bone from each bird, and measuring lead concentration (mg/kg), first via non-destructive XRF and then via destructive inductively coupled plasma mass spectrometry (ICP-MS) and validated the relationship via regression analysis. Portable XRF bone lead measurement demonstrated a strong correlation with ICP-MS results using root-transformed regression (R2 = 0.85). Greater than 92 % of ducks had only background lead exposure (<10 mg/kg). When compared to historical studies in the same species at similar field sites from the 1990s, lead exposure levels were considerably lower, with mean lead concentrations ∼2-fold lower (3.7 c.f. 7.7 mg/kg), and the frequency of birds with severe lead exposure (>20 mg/kg) ∼3-fold lower (2.6 c.f. 7.5 %). Our results confirm that portable XRF is a useful option for measurement of bone lead in Australasian waterbird species. Our findings also demonstrate that a ban on the use of lead shot around 20 years ago has been associated with a substantial reduction in lead exposure in at least one species of waterfowl.

Method to assess the potential magnitude of terrestrial European avian population reductions from ingestion of lead ammunition.

Current estimates of terrestrial bird losses across Europe from ingestion of lead ammunition are based on uncertain or generic assumptions. A method is needed to develop defensible European-specific estimates compatible with available data that does not require long-term field studies. We propose a 2-step method using carcass data and population models. The method estimates percentage of deaths diagnosed as directly caused by lead poisoning as a lower bound and, as an upper bound, the percentage of possible deaths from sublethal lead poisoning that weakens birds, making them susceptible to death by other causes. We use these estimates to modify known population-level annual mortality. Our method also allows for potential reductions in reproduction from lead shot ingestion because reductions in survival and reproduction are entered into population models of species with life histories representative of the most groups of susceptible species. The models estimate the sustainability and potential population decreases from lead poisoning in Europe. Using the best available data, we demonstrate the method on two taxonomic groups of birds: gallinaceous birds and diurnal raptors. The direction of the population trends affects the estimate, and we incorporated such trends into the method. Our midpoint estimates of the reduction in population size of the European gallinaceous bird (< 2%) group and raptor group (2.9-7.7%) depend on the species life history, maximum growth rate, population trend, and if reproduction is assumed to be reduced. Our estimates can be refined as more information becomes available in countries with data gaps. We advocate use of this method to improve upon or supplement approaches currently being used. As we demonstrate, the method also can be applied to individual species of concern if enough data across countries are available.

How contaminated with ammunition-derived lead is meat from European small game animals? Assessing and reducing risks to human health.

Small game animals are generally hunted with lead gunshot which often fragments causing elevated lead concentrations in meat and presenting health risks to frequent consumers and vulnerable groups. We reviewed three decades of European data on lead concentrations in the meat of gamebirds, rabbits and hares across countries with various restrictions on lead gunshot use. Mean meat lead concentrations decreased to a low level in Denmark, the only country in the study with a total ban on lead gunshot use. In contrast, elsewhere in Europe meat lead concentrations increased over time, an unexplained and previously undescribed finding. The only regulatory restriction associated with a decrease in meat lead concentrations was the Danish total ban on lead gunshot use. We calculated an arithmetic mean lead concentration in small game meat of 5.205 ppm w.w. (2011-2021) from across Europe. EU and UK regulators are considering banning lead ammunition for all hunting to protect human and wildlife health. The mean value we found for small game meat (2011-2021) was fourteen times higher than that used in a recent EU-wide risk assessment.

The impact of lead poisoning from ammunition sources on raptor populations in Europe.

Poisoning caused by ingestion of spent lead (Pb) ammunition in food items is a common cause of death of raptors. However, there has been no previous attempt to assess the impact of lead poisoning on populations of raptors throughout Europe or examine how this relates to the prevalence of hunting. We used measurements of lead concentration in the liver from over 3000 raptors of 22 species found dead or dying in the wild in 13 countries and a lead poisoning threshold of 20 ppm (dry weight) to assess the proportion of these in which lead poisoning caused or contributed to death. The prevalence of lead poisoning as a cause of death of raptors varied substantially among European countries and was positively correlated across countries with the reported number of hunters per unit area. Ten species had a non-zero proportion of individuals with concentrations exceeding the lead poisoning threshold ranging between 0.3% and 16.5%. The estimated annual conditional death rate from lead poisoning for these ten species averaged 0.44% (range 0.06-0.85%). Scavenging species feeding regularly on carcasses of game animals,tended to have a high annual probability of death from lead poisoning. So too did some predators which only sometimes scavenge, but prey on frequently hunted birds, such as gamebirds, waterfowl and pigeons, which may contain ingested or embedded lead shot. Small-bodied predators had a low annual probability of death from lead poisoning. Modelling indicated that European populations of adult raptors of the ten focal species averaged 6.0% smaller (range 0.2-14.4%) than they would be without the effects of lead poisoning. A given rate of lead poisoning mortality resulted in greater expected population reductions for species with high annual survival rate and late age at first breeding.

Lead Poisoning among Male Juveniles Due to Illegal Mining: A Case Series from South Africa.

Illegal mining is a major public health and societal concern. Recent scientific evidence indicates elevated blood-lead levels in illegal gold miners and associated communities. Yet, there is little research in this regard from low- to middle-income countries (LMICs), where illegal mining is growing. This case series is extracted from a cross-sectional study of lead exposure in incarcerated juveniles in greater Johannesburg. From survey records (blood-lead levels and questionnaires), three males had elevated blood-lead levels and presented with health conditions and behavioural problems putatively linked with lead poisoning. Based on the record review, all three juveniles were in a secure facility due to illegal mining-related activities. All three cases had high blood-lead levels and demonstrated a tendency toward aggressive or violent behaviour. They also presented with conditions associated with lead poisoning, such as anaemia, respiratory illness, abdominal disorders, and musculoskeletal conditions. Juveniles involved in illegal mining are at risk of exposure to heavy metals such as lead, and there is a need for relevant preventative action and health care programmes in this group.

Major lead exposure from hunting ammunition in eagles from Sweden.

Exposure to lead (Pb) from ammunition in scavenging and raptorial birds has achieved worldwide recognition based on incidences of lethal poisoning, but exposure implies also sublethal levels with potential harmful effects. Background and elevated Pb levels in liver from 116 golden eagles (GE, Aquila chrysaetos) and 200 white-tailed sea eagles (WTSE, Haliaeetus albicilla) from Sweden 2003-2011 are here examined, with supporting data from a previous WTSE report and eagle owl (EO, Bubu bubo) report. GE and WTSE display seasonal patterns, with no Pb level exceeding a generally accepted threshold for subclinical effects during summer but strongly elevated levels from October. Fledged juveniles show significantly lower levels than all other age classes, but reach levels found in older birds in autumn after the start of hunting seasons. Pb levels in EO (non-scavenger) show no seasonal changes and indicate no influence from ammunition, and are close to levels observed in juvenile eagles before October. In all, 15% WTSE and 7% GE were lethally poisoned. In areas with high-exposure to hunting ammunition, 24% of WTSE showed lethal Pb levels, compared to 7% in both eagle species from low-exposure areas. Lethal poisoning of WTSE remained as frequent after (15%) as before (13%) a partial ban on use of Pb-based shotgun ammunition over shallow waters (2002). Pb levels increased significantly in WTSE 1981-2011, in contrast to other biota from the same period. A significant decrease of Pb in WTSE liver occurred below a threshold at 0.25 µg/g (dry weight), exceeded by 81% of the birds. Trend patterns in Pb isotope ratios lend further support to this estimated cut-off level for environmental background concentrations. Pb from spent ammunition affects a range of scavenging and predatory species. A shift to Pb-free ammunition to save wildlife from unnecessary harm is an important environmental and ethical issue.

Role of dietary curcumin against waterborne lead toxicity in common carp Cyprinus carpio.

Curcumin exhibited numerous key activities including antioxidant, anti-inflammatory, and immunostimulatory effects in fish. This study evaluated the protective effects of curcumin (CUR) against lead (Pb)-induced toxicities in fish. Healthy Cyprinus carpio L. individuals were segregated into control, Pb only, Pb+CUR, and CUR only groups. Pb groups were exposed to 1 mg L -1 of Pb, and CUR groups were fed a basal diet supplemented with 15 g kg-1 of CUR. After eight weeks, growth performance, Pb accumulation in tissues, various haemato-biochemical parameters, immune responses, and cytokine gene expression were measured. Dietary CUR effectively decreased Pb accumulation in tissues and increased the survival of Pb-exposed fish. Co-treatment with Pb and CUR reversed alterations in haemato-biochemical parameters, ameliorated Pb-induced oxidative stress, enhanced immune responses, and restored intestinal enzymatic activities. Dietary CUR reversed changes in intestinal microbiota in Pb-exposed fish. Pb-induced upregulation of NF-κBp65 and HSP70 was inhibited by dietary CUR. CUR supplementation upregulated the mRNA levels of SOD, Nrf2, IL-10, and CYP450 1A and attenuated Pb-induced degradation of I κB-α mRNA levels. Overall, CUR antagonizes Pb-induced negative impacts in fish. Thus, dietary CUR had several beneficial effects on immune responses, decreased Pb accumulation in tissues, and reversed Pb-induced oxidative stress in fish. Therefore, CUR plays a protective role in Pb-induced immune toxicity in fish, and, as such, may be suitable as an aqua feed additive for use in carp aquaculture.

Application of Stable Isotopic Ratio Analysis to Identify the Cause of Acute Versus Chronic Lead Poisoning of a Tundra Swan (Cygnus columbianus): A Case Study.

We utilized lead (Pb) stable isotopes to identify the source of acute Pb poisoning in a Tundra Swan (Cygnus columbianus) and evaluated overall Pb exposure. Upon necropsy, we obtained samples of blood, liver, kidney, heart, thigh, breast, femur, and metallic objects (i.e., fishing sinker, spring and swivels) from the gizzard for Pb isotopic analysis. Pb isotope ratios of blood and soft tissues were essentially identical to the Pb ratios of the sinker, the likely source of acute poisoning. The spring and swivels had lower Pb content and ratios distinct from tissue, suggesting no significant contribution to poisoning. Femur Pb isotopic composition was the most distinct biological sample and indicative of a combination of sources. These results demonstrate isotopic analysis as a viable method for determining the source of acute Pb poisoning, and that Pb isotope ratios in bone most likely record a lifetime-averaged metric of Pb exposure.

Saikosaponin a attenuates lead-induced kidney injury through activating Nrf2 signaling pathway.

Saikosaponin a (SSa), a triterpene saponin extracted from Bupleurumfalcatum L., has been reported to have anti-inflammatory activity. In the present study, we investigated the effects of SSa on lead-induced kidney injury in common carp. Kidney histological changes were detected by H&E staining. The levels of TNF-α, IL-1ß, MPO, MDA, GSH, and SOD activity were also measured. Furthermore, the NF-κB and Nrf2 signaling pathways were tested by western blot analysis. The results showed that lead-induced kidney histological change was attenuated by SSa. Lead-induced TNF-α, IL-1ß, MPO, and MDA production were also suppressed by SSa. Meanwhile, lead could decrease GSH level and SOD activity and the decreases were inhibited by SSa. Furthermore, we found SSa significantly inhibited lead-induced NF-κB translocation. In addition, the expression of Nrf2 and HO-1 were increased by the treatment of SSa and Keap1 expression was decreased by SSa. In conclusion, this study indicated that SSa inhibited lead-induced kidney injury in carp through suppressing inflammatory and oxidative responses, and the mechanism may be involved in the inhibition of NF-κB and activation of Nrf2 signaling pathway.

Integrating active and passive monitoring to assess sublethal effects and mortality from lead poisoning in birds of prey.

The ingestion of lead (Pb) ammunition is the most important exposure pathway to this metal in birds and involve negative consequences to their health. We have performed a passive monitoring of Pb poisoning in birds of prey by measuring liver (n = 727) and blood (n = 32) Pb levels in individuals of 16 species found dead or sick in Spain between 2004 and 2020. We also performed an active monitoring by measuring blood Pb levels and biomarkers of haem biosynthesis, phosphorus (P) and calcium (Ca) metabolism, oxidative stress and immune function in individuals (n = 194) of 9 species trapped alive in the field between 2016 and 2017. Passive monitoring results revealed some species with liver Pb levels associated with severe clinical poisoning (>30 µg/g d.w. of Pb): Eurasian griffon vulture (27/257, 10.5%), red kite (1/132, 0.8%), golden eagle (4/38, 10.5%), and Northern goshawk (1/8, 12.5%). The active monitoring results showed that individuals of bearded vulture (1/3, 33.3%), Eurasian griffon vulture (87/118, 73.7%), Spanish imperial eagle (1/6, 16.7%) and red kite (1/18, 5.6%) had abnormal blood Pb levels (>20 µg/dL). Blood Pb levels increased with age, and both monitoring methods showed seasonality in Pb exposure associated with a delayed effect of the hunting season. In Eurasian griffon, blood Pb concentration was associated with lower δ-ALAD activity in blood and P levels in plasma, and with higher blood lipid peroxidation and plasma carotenoid levels in agreement with other experimental and field studies in Pb-exposed birds. The study reveals that Pb poisoning is a significant cause of death and sublethal effects on haem biosynthesis, P metabolism and oxidative stress in birds of prey in Spain.

Diagnosis and Treatment of Heavy Metal Toxicosis in Six Waldrapp Ibis (Geronticus eremita).

Six adult Waldrapp ibis (Geronticus eremita) were presented with heavy metal toxicosis secondary to paint chip ingestion after being placed in an indoor confinement enclosure because of a regional highly pathogenic avian influenza epornitic and government restrictions. Four of the ibis developed clinical disease signs between 45 and 64 days of confinement, including weakness, lethargy, inability to fly, low body condition, and bilateral wing droop while 2 birds appeared to remain normal during that period. Common clinicopathological findings included elevated plasma aspartate aminotransferase, creatine kinase, glutamate dehydrogenase, and bile acids values. Erythrocyte morphological changes included prominent hypochromasia, decreased size of polychromatophils with occasional cytoplasmic stippling, abnormal shapes, and D cells. Whole-body radiographic imaging revealed particulate radio-opaque material in the ventriculus of all affected birds. One bird died before chelation therapy could be instituted and heavy metal testing of the liver revealed increased concentrations of lead and zinc. Lead toxicosis was diagnosed in the 5 remaining birds by plasma lead analysis. These 5 birds were hospitalized for an average of 40 days and treated with sodium calcium edetate and fluid therapy, which resulted in resolution of the toxicosis. Plasma biochemistry, complete blood counts, and blood lead values were performed repeatedly on all 5 birds and were used to guide therapy and monitor treatment response. This case series emphasizes the importance of early contingency planning and reviews the treatment of lead toxicosis in birds with a focus on long-term clinical monitoring and hematology.

Current situation regarding lead exposure in birds in Japan (2015-2018); lead exposure is still occurring.

Birds of a number of species have died as a result of lead (Pb) poisoning, including many Steller's sea eagles (Haliaeetus pelagicus) and white-tailed sea eagles (Haliaeetus albicilla) in Hokkaido, the northernmost island of Japan. To address this issue, the use of any type of Pb ammunition for hunting of large animals was prohibited in Hokkaido in 2004. However, Pb poisoning is still being reported in this area, and there are few regulations regarding the use of Pb ammunition in other parts of Japan, where it has been reported that eagles and water birds have been exposed to Pb. This study was performed to accurately determine the current level of Pb exposure of birds found dead in the field or dead in the wild bird centers in Japan (June 2015-May 2018) and to identify the sources of Pb. Pb exposure was found to still be occurring in raptors and water birds in various parts of Japan. Twenty-six point five % and 5.9% of the recorded deaths of Steller's sea eagles and white-tailed sea eagles, respectively, were found to have been poisoned by Pb. In addition, Pb isotope ratio analysis showed that both Pb rifle bullets and Pb shot pellets cause Pb exposure in birds, and these endangered eagles are also exposed to Pb in Hokkaido due to the illegal use of Pb ammunition. Changing to Pb-free ammunition, such as copper (Cu) rifle bullets, steel shot pellets, or bismuth shot pellets, will be essential for the conservation of avian species in Japan.