Concentration of metals and metalloids in livers of birds of various foraging guilds collected during the autumn migration period in Poland.

During migration, birds explore various habitats at stopover sites that differ in food resources and contamination levels. In this study, hepatic concentrations of 21 elements (metals and metalloids) in 11 species of birds, representing various foraging habitats (such as aquatic, aquatic/terrestrial, and terrestrial) and migration modes (migratory and sedentary) representing various foraging guilds (omnivores, piscivores, and molluscivores), were analyzed. The samples (N = 84) were collected during the autumn migration period in Poland. The concentrations of elements determined in this study exhibited high inter-species variability, reflecting the diversity in contamination levels depending on food resources used by specific bird groups. Many of the investigated individuals from different species showed exceeded levels of subclinical toxicity and moderate clinical poisoning due to Cd and Hg. Higher concentrations of As, Hg, and Ba and lower V concentrations were found in migratory birds as compared to sedentary birds. Species foraging in terrestrial habitat had different concentrations of some elements compared to aquatic and aquatic/terrestrial species. Some specific inter-species differences in hepatic elemental concentrations were found. Differences in elemental concentrations among various groups can primarily be attributed to their foraging guilds, with certain elements, particularly As, V, and Hg, playing a significant role in the dissimilarity of elemental concentrations between foraging habitat groups and migratory mode groups. The data collected confirmed the limited ability of As to enter ecosystem pathways. The results of this study contribute to understanding the year-round exposure of migratory birds to environmental contamination, which can have carry-over effects on their performance in wintering and breeding grounds.

A novel type of biochar from chitinous Hermetia illucens waste with a built-in stimulating effect on plants and soil arthropods.

The breeding of insects generates waste in the form of insect excrement and feed residues. In addition, a specific chitinous waste in the form of insect larvae and pupae exuvia is also left. Recent research tries to manage it, e.g., by producing chitin and chitosan, which are value-added products. The circular economy approach requires testing new, non-standard management methods that can develop products with unique properties. To date, the possibility of biochar production from chitinous waste derived from insects has not been evaluated. Here we show that the puparia of Hermetia illucens are suitable for biochar production, which in turn exhibits original characteristics. We found that the biochars have a high nitrogen level, which is rarely achievable in materials of natural origin without artificial doping. This study presents a detailed chemical and physical characterization of the biochars. Moreover, ecotoxicological analysis has revealed the biochars' stimulation effect on plant root growth and the reproduction of the soil invertebrate Folsomia candida, as well as the lack of a toxic effect on its mortality. This predisposes these novel materials with already built-in stimulating properties to be used in agronomy, for example as a carriers for fertilizers or beneficial bacteria.

Comorbidities and concentration of trace elements in livers of European bison from Bieszczady Mountains (Poland).

European bison is a species for which health monitoring is essential in conservation activities. So far, little research has been carried out on the concentration of elements in this species. Most previous studies did not associate the concentration of elements with susceptibility to diseases. In this study we investigate the relationship between comorbidities in European bison and concentrations of a wide spectrum of elements in the liver. Samples were collected during the monitoring of the European bison population in Bieszczady (southeast Poland) over the 2020-2022 period. Each individual was also visually inspected by a veterinarian in the field for the presence of lesions as a part of a post-mortem examination. The animals were divided into 3 groups: group A-one type of clinical sign; group B-two types of clinical signs; group C-three or more types of clinical signs. The ICP-OES method was applied to assess the concentration of 40 elements in livers. Discriminant analysis showed clear differences between the mineral status of individuals in the groups with one, two, and at least three types of clinical signs. Detailed analysis of selected elements showed that, in the case of eight elements, there was a relationship with age, sex, or comorbidities. Cu, Se, and Zn showed significant differences in relation to comorbidities, but only Cu concentration was lower when the frequency of lesions was higher. We concluded that in research on the mineral status of the population, apart from the availability of trace elements in the environment, the health condition of the studied individuals should also be considered. However, inferring the mineral status of the population on the basis of randomly obtained samples from dead individuals may give an incomplete view of the population, especially in the case of species susceptible to diseases, such as European bison.

Imidacloprid Pesticide Causes Unexpectedly Severe Bioelement Deficiencies and Imbalance in Honey Bees Even at Sublethal Doses.

Pesticides impair honeybee health in many ways. Imidacloprid (IMD) is a pesticide used worldwide. No information exists on how IMD impact the bees' body bioelement balance, which is essential for bee health. We hypothesized that IMD disturbs this balance and fed the bees (in field conditions) with diets containing 0 ppb (control), 5 ppb (sublethal considered field-relevant), and 200 ppb (adverse) doses of IMD. IMD severely reduced the levels of K, Na, Ca, and Mg (electrolytic) and of Fe, Mo, Mn, Co, Cu, Ni, Se, and Zn, while those of Sn, V, and Cr (enzymatic) were increased. Levels of P, S, Ti, Al, Li, and Sr were also decreased, while only the B content (physiologically essential) was increased. The increase in Tl, Pb, and As levels (toxic) was alarming. Generally, IMD, even in sublethal doses, unexpectedly led to severe bioelement malnutrition in 69% of bioelements and to a stoichiometric mismatch in the remaining ones. This points to the IMD-dependent bioelement disturbance as another, yet unaccounted for, essential metabolic element which can interfere with apian health. Consequently, there is a need for developing methods of bioelement supplementation of the honey bee diet for better preventing bee colony decline and protecting apian health status when faced with pesticides.

Assessing Bone Health Status and Eggshell Quality of Laying Hens at the End of a Production Cycle in Response to Inclusion of a Hybrid Rye to a Wheat-Corn Diet.

The objective of this study was to evaluate whether there are negative effects of the partial replacement of white corn with rye along with xylanase supplementation on overall bone quality, eggshell mineralization, and mechanical strength in laying hens. From the 26th week of life, ISA Brown laying hens were fed either a wheat-corn diet or a diet containing 25% rye, with or without xylanase. The experimental period lasted for 25 weeks, until birds reached their 50th week of age, after which bone and eggshell quality indices were assessed. Eggshell thickness and eggshell Ca content of eggs from rye-fed hens were improved by xylanase supplementation. No differences in the mechanical properties of the eggshells were observed between treatments, except for the diet-dependent changes in egg deformation. Rye inclusion had no effect on the mechanical properties of bone. Xylanase supplementation, irrespective of the diet, had a positive effect on bone strength and increased tibia Ca content, as well as the content of several microelements. Hence, hybrid rye combined with wheat can replace 25% of corn in layer diets without compromising shell quality or bone mineral content. Xylanase supplementation in these diets is recommended since its inclusion improves both bone strength and quality.

Soluble Extracellular Polymeric Substances Produced by Parachlorella kessleri and Chlorella vulgaris: Biochemical Characterization and Assessment of Their Cadmium and Lead Sorption Abilities.

In the present study, the potential of lead and cadmium removal by the extracellular polymeric substances (EPS) produced from Parachlorella kessleri and Chlorella vulgaris were investigated. Carbohydrates were the dominant components of EPS from both analyzed species. The contents of reducing sugars, uronic acids, and amino acids were higher in EPS synthesized by C. vulgaris than in EPS from P. kessleri. The analysis of the monosaccharide composition showed the presence of rhamnose, mannose and galactose in the EPS obtained from both species. The ICP-OES (inductively coupled plasma optical emission spectrometry) analyses demonstrated that C. vulgaris EPS showed higher sorption capacity in comparison to P. kessleri EPS. The sorption capacity of C. vulgaris EPS increased with the increase in the amount of metal ions. P. kessleri EPS had a maximum sorption capacity in the presence of 100 mg/L of metal ions. The FTIR analysis demonstrated that the carboxyl, hydroxyl, and carbonyl groups of EPS play a key role in the interactions with metal ions. The present study showed C. vulgaris EPS can be used as a biosorbent in bioremediation processes due to its biochemical composition, the presence of significant amounts of negatively charged uronic acids, and higher sorption capacity.

Physicochemical Properties and Surface Characteristics of Ground Human Teeth.

Enamel, dentin and cementum apatite has a complex composition. The lack of complete reports on the chemical composition of all tooth tissues together and the need to create a modern biomaterial that reproduces the correct ratio of individual tooth mineral components prompted the authors to undertake the research. A detailed evaluation of the micro- and macro-elements of tooth powder, using various methods of chemical analysis was conducted. All four groups of human sound teeth were crushed using the grinder. A fine powder was implemented for the FTIR (Fourier Transform Infrared Spectroscopy), ICP (Inductively Coupled Plasma Optical Emission Spectometry) and for the potentiometric titration, SEM and mercury porosimetry analyses. The obtained studies indicate that there is no significant correlation in chemical composition between the different teeth types. This proves that every removed, crushed tooth free of microorganisms can be a suitable material for alveolar augmentation. It is essential to know the chemical profiles of different elements in teeth to develop a new class of biomaterials for clinical applications.

Dietary Intake of Toxic Heavy Metals with Major Groups of Food Products-Results of Analytical Determinations.

Food contains a complex matrix of various substances, including essential nutrients, non-nutritive substances, and toxins, including metals. The main purpose of the study was to evaluate the contribution of major groups of food products to an overall intake of toxic heavy metals (Cd, Pb, Hg, and Ni) using a combination of the 24-dietary recall technique, the ICP-OES (Inductively Coupled Plasma Optical Emission Spectrometry) method, and chemometric tools. The obtained results reveal that there is a high potential risk of developing nephrotoxicity through the dietary intake of Pb in the case of both genders. The dietary intake determined for other elements (Cd, Hg, and Ni) was far below the limits established by European Food Safety Authority (EFSA). Principal Component Analysis (PCA) supported analytical determinations and revealed that cereals and vegetables were major contributors to a total intake of Cd (39.6 and 17.4% of the total exposure, respectively), Ni (40.4 and 19.3%), and Hg (16.8 and 19.6%), while water and beverages were major dietary sources of Pb (31% of the total daily intake). In contrast, eggs, fats and oils, and milk and dairy products provided the smallest amounts of Cd, Pb, and Ni. Despite containing high amounts of Hg, considering very low consumption, fish were not found to be an important source of this element.

Changes in the Intestinal Histomorphometry, the Expression of Intestinal Tight Junction Proteins, and the Bone Structure and Liver of Pre-Laying Hens Following Oral Administration of Fumonisins for 21 Days.

Fumonisins (FB) are metabolites found in cereal grains (including maize), crop products, and pelleted feed. There is a dearth of information concerning the effects of FB intoxication on the intestinal histomorphometry, the expression of intestinal tight junction proteins, and the bone structure and liver in pre-laying hens. The current experiment was carried out on hens from the 11th to the 14th week of age. The hens were orally administered an extract containing fumonisin B1 (FB1) and fumonisin B2 (FB2) at doses of 0.0 mg/kg b.w. (body weight), 1.0 mg/kg b.w., 4.0 mg/kg b.w., and 10.9 mg/kg b.w. for 21 days. Following FB intoxication, the epithelial integrity of the duodenum and jejunum was disrupted, and dose-dependent degenerative changes were observed in liver. An increased content of immature collagen was observed in the bone tissue of FB-intoxicated birds, indicating intensified bone turnover. A similar effect was observed with regards to the articular cartilage, where enhanced fibrillogenesis was observed mainly in the group of birds that received the FB extract at a dose of 10.9 mg/kg b.w. In conclusion, FB intoxication resulted in negative structural changes in the bone tissue of the hens, which could result in worsened bone mechanics and an increase in the risk of bone fractures. Fumonisin administration, even at a dose of 1.0 mg/kg b.w., can lead to degradation of the intestinal barrier and predispose hens to intestinal disturbances later in life.

Essential differences in the mineral status of free-ranging European bison Bison bonasus populations in Poland: The effect of the anthroposphere and lithosphere.

Tracking the concentration of elements in the organs of endangered animal species can provide significant knowledge about environmental hazards and allows more targeted protective measures to be taken. In this paper the differences in concentrations of 27 elements in the livers of free-ranging European bison were studied. The ICP-OES method was applied to study the four main Polish populations, which live in lowland (Bialowieska, Knyszynska and Borecka forests) and mountain (Bieszczady) habitats. The study was based on a unique and relatively large amount of comparative material. It was found that European bison populations that live in different locations differ from each other in terms of the hepatic concentrations of 17 of the elements studied in this work. The mountain population in Bieszczady was most distinctive due to the higher concentrations of Ba, Ca, Cd, Se, and the lower concentrations of Mo and V (compared to the other populations). Additionally, even geographically close lowland populations sometimes have different hepatic concentrations of particular elements. The sex and age of individuals have a weak effect on mineral status. Although the lithosphere is the primary factor that affects the availability of elements for the European bison, the soil parameters and biosphere can significantly affect the availability of elements, e.g. by accumulation. It was also shown that the anthroposphere plays a role, but this is mainly through agriculture as feeding on arable fields significantly changes the mineral status of European bison. The need for further research, especially on Cd, Se and Cu, was indicated. It was concluded that toxic elements or deficiencies in essential elements in the populations of the studied species should be considered locally as this would allow better understanding of the health condition of individual populations.

Contribution of Major Groups of Food Products to the Daily Intake of Selected Elements-Results from Analytical Determinations Supported by Chemometric Analysis.

Food is a major source of minerals for humans. The main objective of this study was to determine the intake level of 10 essential macro- (Na, K, Ca, and Mg) and trace elements (Cu, Zn, Mn, Fe, Cr, and Se) with major food groups among young adults. Dietary intake of elements was evaluated using the 24-h dietary recall technique in combination with F-AASand ICP-OES methods. A very high intake of sodium and a very low intake of calcium, combined with inappropriate sodium/potassium ratio, may be harmful to the health of the population. Dietary intake of trace elements was within the range of reference values in the subjects, with cereals being the major source of a majority of those elements, while meat (38% for Na), vegetables (25% for K), and milk products (75% for Ca) were the main contributors to the daily dietary intake of macroelements. PCA revealed several visible trends in the datasetAmong men, the intake of Zn, Cr Na and K was significantly correlated with the consumption of meat and vegetables, whereas Mg, Se, Fe and Cu with cereals and water and beverages. Among women, the intake of Mg was significantly correlated with the consumption of meat and vegetables.

Effect of Fe3O4 nanoparticles on germination of seeds and concentration of elements in Helianthus annuus L. under constant magnetic field.

The aim of the study was to investigate the effect of the Fe3O4 nanoparticles (Fe-NPs) on the germination of sunflower seeds, early growth of seedlings and the concentration of selected elements in seedlings. The influence of constant magnetic fields in systems with and without Fe-NPs was investigated. Experiments were done on seeds subjected to germination under constant magnetic field (0 (control), 5, 25 and 120 mT) for 7 days in the presence of solution containing 0, 50 or 500 ppm Fe-NPs. No significant effect of Fe-NPs and the magnetic field on germination of seeds and the growth of seedlings has been demonstrated. In most cases, a decrease in germination parameters was observed. For the majority of samples the relative decrease in the concentrations of elements was demonstrated mainly for samples without Fe-NPs. Interestingly, a significant decrease in the concentrations of trivalent (including iron - Fe) and toxic elements in samples containing Fe-NPs in relation to control samples was observed. The authors suggest that in this case the binding (adsorption) of these elements in the roots and seeds of the sunflower by Fe-NPs took place. This explains the lower iron content in seedlings than in seeds prior to sowing.

Agro-industrial by-product in photoheterotrophic and mixotrophic culture of Tetradesmus obliquus: Production of ω3 and ω6 essential fatty acids with biotechnological importance.

In recent years, researchers have highlighted the role of low cost-efficient agro-industrial by-products used as supplements in algal culture media. The aim of the study was to identify and characterize the basic metabolic pathways in Tetradesmus obliquus cells induced by supplementation with beet molasses in photoheterotrophic and mixotrophic culture conditions. To assess the impact of the nutritional strategy in unicellular algae, growth curves were plotted and lipid, carbohydrate, and protein levels were determined. Fourier Transform Infrared Spectroscopy was applied to measure the Tetradesmus obliquus cell composition. Additionally, the C16-C18 fatty acid profile of Tetradesmus obliquus was determined by gas chromatograph/mass spectrometry. The switch from autotrophy to photoheterotrophy and mixotrophy contributes to shortening of the adaptation growth phase. The highest protein content was obtained in the mixotrophic growth. This study has demonstrated high content of 18:1, cisΔ9, 18:2, cisΔ9,12, ω6, and 18:3, cisΔ9,12,15, ω3 in photoheterotrophic and mixotrophic culture conditions. High levels of proteins and essential fatty acids make Tetradesmus obliquus cell biomass important for human and animals health.

Hermetia illucens exhibits bioaccumulative potential for 15 different elements - Implications for feed and food production.

A new branch of the insect-based food and feed industry is intensively developing all over the world, the best proof of which is the recent change in legislation at the European level allowing the use of insect protein in the production of feed for aquaculture animals. Previous publications have proven that some heavy metals can be bioaccumulated in fly H. illucens (black soldier fly), even when the insect is raised on optimal feed with an acceptable heavy metal content. The purpose of this study was to determine the bioaccumulation potential of H. illucens in relation to micro- and macroelements, toxic elements and for the first time, selected non-essential elements from optimal feed. Our results showed that bioaccumulation of Ba, Bi, Cu, Fe, Hg, Mg, Mo, Se and Zn occurred in all stages of insect development and in puparia, while bioaccumulation of Al, As, Co, K, Pb and Si was not found. The highest bioaccumulation factors were obtained for Ca and Mn in puparia - 38 and 21 respectively. In addition, Ca, Cd, Ga, Mn, P and S were bioaccumulated only in some developmental stages of the insect. The results are discussed in the context of the safety of feed and food production from H. illucens.

Liquid Anaerobic Digestate as a Source of Nutrients for Lipid and Fatty Acid Accumulation by Auxenochlorella Protothecoides.

In recent years, there has been growing interest in the biomass of unicellular algae as a source of valuable metabolites. The main limitations in the commercial application of microbial biomass are associated with the costs of production thereof. Maize silage is one of the main substrates used in biogas plants in Europe. The effects of sterilized agricultural liquid digestate (LD) from methane fermentation of maize silage on the growth rates, macro and micronutrient removal efficiency, lipid content, and fatty acid profile in Auxenochlorella protothecoides were investigated. The results indicate that A. prothecoides can proliferate and accumulate lipids with simultaneous reduction of nutrients in the 1:20 diluted liquid digestate. The rate of nitrogen and phosphorus removal from the liquid digestate was 79.45% and 78.4%, respectively. Cells growing in diluted liquid digestate exhibited the maximum lipid content, i.e., 44.65%. The fatty acid profile of A. prothecoides shows a decrease in the content of linolenic acid by 20.87% and an increase in oleic acid by 32.16% in the LD, compared with the control. The liquid digestate changed the content of monounsaturated fatty acids and polyunsaturated fatty acids. The cells of A. protothecoides growing in the liquid digestate were characterized by lower PUFA content and higher MUFA levels.

The preliminary studies on protein profile in retained and not retained foetal membranes in heavy draft mares.

Protein profile of the placenta expresses its function and maintenance. Any alterations can be reflected in qualitative and quantitative changes in this profile. The aim of the present study was the evaluation of protein profile in the placenta of mares suffering from the retention of foetal membranes (FMR) by two separation methods and the comparison with physiologically released tissues. Placentas from 14 healthy, heavy draft mares were collected immediately after the expulsion of newborn. Tissues after homogenization and staining with fluorescent dyes were subjected to electrophoretic as well as chromatographic separation. Electrophoretic gels were statistically analysed for the presence and abundance of examined proteins, while some proteins were identified in chromatographic fractions. Out of 248 spots detected in endometrium, 38 differed significantly between FMR and control animals, while in allantochorion, respective values reached 241 and 27 spots (p < .05). Among identified proteins that expressed higher abundance in endometrium of FMR mares than control animals were prostaglandin reductase, dehydrogenase/reductase SDR family, and placental growth factor. These proteins are involved in regulation of parturition. Additionally, the following proteins responsible for physiological activity of a cell-guanine methyl transferase, aspartyl/asparaginyl beta-hydroxylase and GTP-binding protein, were identified. These proteins expressed higher abundance in allantochorion of FMR mares than in controls. This preliminary study confirmed the disturbances in protein pattern between foetal membranes in FMR and healthy mares. Further qualitative and quantitative experiments are necessary to deepen our knowledge on the mechanisms of the retention of foetal membranes in mares.

Fungal (1 → 3)-α-d-glucans as a new kind of biosorbent for heavy metals.

Although the fungal ability to bind heavy metals has been known for many years, it was not determined which component is responsible for this process. The aim of the study was to isolate (1 → 3)-α-d-glucans from various fungi, select the most efficient compound in the biosorption of heavy metals, and determine its characteristics. The best α-glucan for treatment of aqueous solutions from heavy metals (Ni2+, Cd2+, Zn2+, Pb2+) is the polymer isolated from Shiitake, especially its SH 37 variety. Using various techniques, it was confirmed that the studied polysaccharide consists mainly of glucose molecules (75.9%), i.e. glucose α-anomers linked by (1 → 3)-linkage (86%). The well-developed surface, low crystallinity, and the large number of -OH groups provide this polymer with good sorption properties. Probably, the main mechanism of metal uptake is metabolism-independent process that depends on the content and spatial structure of the glucans present in their cell wall.

The effect of cadmium exposition on the structure and mechanical properties of rat incisors.

Alterations in the structure and mechanical properties of teeth in adult Wistar rats exposed to cadmium were investigated. Analyses were conducted on two sets of incisors from female and male specimens, that were intoxicated with cadmium (n = 12) or belonged to the control (n = 12). The cadmium group was administered with CdCl2 dissolved in drinking water with a dose of 4mg/kgbw for 10 weeks. The oral intake of cadmium by adult rats led to the range of structural changes in enamel morphology and its mechanical features. A significant increase of cadmium levels in the teeth in comparison to the control, a slight shift in the colour and reduction of pigmented enamel length, higher surface irregularity, a decrease of hydroxyapatite crystals size in the c-axis and simultaneous increase in pigmented enamel hardness were observed. The extent of these changes was sex-dependent and was more pronounced in males.

Maternal HMB treatment affects bone and hyaline cartilage development in their weaned piglets via the leptin/osteoprotegerin system.

It has been demonstrated in animal studies that prenatal administration of ß-hydroxy-ß-methylbutyrate (HMB, metabolite of leucine) influences general growth and mechanical endurance of long bones in newborn offspring in sex-dependent manner. The present experiment was conducted to evaluate the effect of HMB treatment of pregnant sows on bone development in offspring at weaning. From 70th day until the 90th day of gestation, sows received either a basal diet (n = 12) or the same diet supplemented with HMB (n = 12) at the dose of 0.2 g/kg of body weight/day. Femora obtained from six males and females in each group weaned at the age of 35 days were examined. Maternal HMB treatment significantly enhanced body weight and changed bone morphology increasing femur mechanical strength in both sexes. Maternal HMB supplementation also elevated bone micro- and macroelement concentrations and enhanced content of proteoglycans in articular cartilage. Based on the obtained results, it can be concluded that maternal HMB supplementation in the mid-gestation period significantly accelerated bone development in both sexes by upregulation of a multifactorial system including leptin and osteoprotegerin. However, the sex (irrespective of the HMB treatment) was the factor which influenced the collagen structure in cartilages and trabecular bone, as demonstrated both by the Picrosirius red staining and performed analysis of thermal stability of collagenous tissues. The structural differences in collagen between males and females were presumably related to a different collagen maturity. No studies conducted so far provided a detailed morphological analysis of bone, articular cartilage, growth plate and the activities of the somatotropic and pituitary-gonadal axes, as well as leptin/osteoprotegerin system in weaned offspring prenatally treated with HMB. This study showed also the relationship between the maternal HMB treatment and bone osteometric and mechanical traits, hormones, and growth and bone turnover markers such as leptin, osteoprotegerin and insulin-like growth factor-1.

Inter-species and inter-colony differences in elemental concentrations in eggshells of sympatrically nesting great cormorants Phalacrocorax carbo and grey herons Ardea cinerea.

We compared the concentrations of 17 heavy metals and essential elements in post-hatching eggshells of two waterbirds, the obligate piscivorous great cormorant Phalacrocorax carbo (GCM) and the more omnivorous grey heron Ardea cinerea (GHR), breeding sympatrically in eight mixed colonies in Poland. We found significant inter-species and inter-colony differences in the levels of most of the elements. GHR had significantly higher concentrations of Al, which can be explained by its very low stomach pH: an acidic environment favours the release of Al compounds. Differences in Mn, Ni, Cu, Se and Hg concentrations can be attributed to the various contributions of fish and other aquatic organisms to the diet, and to the exploration of different habitats (GCM exclusively aquatic, GHR a wider range) and microhabitats (GCM, in contrast to wading GHR, dive for food, exploring the whole depth range of water bodies), differently exposed to contamination by those elements from sediments. Inter-colony differences were related to the level of industrialisation. We recorded higher levels of some elements in the eggshells (Fe, Mn in both species and Cr, Ni and Zn in GCM) collected in industrialised areas, which may be associated with the negative environmental impact of industrial areas.