An integrative and multi-indicator approach for wildlife health applied to an endangered caribou herd.

Assessing wildlife health in remote regions requires a multi-faceted approach, which commonly involves convenient samplings and the need of identifying and targeting relevant and informative indicators. We applied a novel wildlife health framework and critically assessed the value of different indicators for understanding the health status and trends of an endangered tundra caribou population. Samples and data from the Dolphin and Union caribou herd were obtained between 2015 and 2021, from community-based surveillance programs and from captured animals. We documented and categorized indicators into health determinants (infectious diseases and trace elements), processes (cortisol, pathology), and health outcomes (pregnancy and body condition). During a recent period of steep population decline, our results indicated a relatively good body condition and pregnancy rates, and decreasing levels of stress, along with a low adult cow survival. We detected multiple factors as potential contributors to the reduced survival, including Brucella suis biovar 4, Erysipelothrix rhusiopathiae and lower hair trace minerals. These results remark the need of targeted studies to improve detection and investigations on caribou mortalities. We also identified differences in health indicators between captured and hunter sampled caribou, highlighting the importance of accounting for sampling biases. This integrative approach that drew on multiple data sources has provided unprecedented knowledge on the health in this herd and highlights the value of documenting individual animal health to understand causes of wildlife declines.

Improved RP-HPLC separation of Hg²âº and CH3Hg⁺ using a mixture of thiol-based mobile phase additives.

Hg(2+) and CH(3)Hg(+) are frequently encountered in the environment either as free ions or complexed with organic matter, such as humic acids. The majority of the reported HPLC-based separations of environmental mercury species, however, separate Hg(2+) from CH(3)Hg(+) in which the former species elutes close to the void volume. To detect mercury-species in environmental waters that may have so far escaped detection, a separation method is needed that sufficiently retains both Hg(2+) and CH(3)Hg(+). One way to develop such a method is to increase the retention of Hg(2+) and CH(3)Hg(+) using existing HPLC separations. We here report on the improvement of a previously reported RP-HPLC-based separation of Hg(2+) and CH(3)Hg(+) that employed a 100 % aqueous mobile phase [10 mM L-cysteine (Cys) in 50 mM phosphate buffer (pH 7.5)]. To increase the retention of Hg(2+), Cys was replaced by the comparatively more hydrophobic N-acetylcysteine (N-Cys). To achieve a compromise between an increased retention of Hg(2+) and its baseline separation from CH(3)Hg(+) in the shortest possible analysis time, the retention behavior of both mercurials was investigated on two RP-HPLC columns with mobile phases that contained mixtures of Cys and N-Cys in which the overall thiol concentration was maintained at 10 mM. An optimal separation of both mercurials could be achieved in ∼540 s using a Gemini C(18) HPLC column (150 × 4.6 mm I.D.) and a mobile phase comprised of 7.5 mM N-Cys and 2.5 Cys in 50 mM phosphate buffer (pH 7.4). Coupling the developed HPLC separation with an inductively coupled plasma mass spectrometer should allow one to detect mercury species other than Hg(2+) and CH(3)Hg(+) in environmental waters. The detection of such species is critical to better understand the mobilization of mercury species from natural and anthropogenic pollution sources.

Determination of chlorophenols in water samples using simultaneous dispersive liquid-liquid microextraction and derivatization followed by gas chromatography-electron-capture detection.

Simultaneous dispersive liquid-liquid microextraction (DLLME) and derivatization combined with gas chromatography-electron-capture detection (GC-ECD) was used to determine chlorophenols (CPs) in water sample. In this derivatization/extraction method, 500 microL acetone (disperser solvent) containing 10.0 microL chlorobenzene (extraction solvent) and 50 microL acetic anhydride (derivatization reagent) was rapidly injected by syringe in 5.00 mL aqueous sample containing CPs (analytes) and K(2)CO(3) (0.5%, w/v). Within a few seconds the analytes derivatized and extracted at the same time. After centrifugation, 0.50 microL of sedimented phase containing enriched analytes was determined by GC-ECD. Some effective parameters on derivatization and extraction, such as extraction and disperser solvent type and their volume, amount of derivatization reagent, derivatization and extraction time, salt addition and amount of K(2)CO(3) were studied and optimized. Under the optimum conditions, enrichment factors and recoveries are in the range of 287-906 and 28.7-90.6%, respectively. The calibration graphs are linear in the range of 0.02-400 microg L(-1) and limit of detections (LODs) are in the range of 0.010-2.0 microg L(-1). The relative standard deviations (RSDs, for 200 microg L(-1) of MCPs, 100 microg L(-1) of DCPs, 4.00 microg L(-1) of TCPs, 2.00 microg L(-1) of TeCPs and PCP in water) with and without using internal standard are in the range of 0.6-4.7% (n=7) and 1.7-7.1% (n=7), respectively. The relative recoveries of well, tap and river water samples which have been spiked with different levels of CPs are 91.6-104.7, 80.8-117.9 and 83.3-101.3%, respectively. The obtained results show that simultaneous DLLME and derivatization combined with GC-ECD is a fast simple method for the determination of CPs in water samples.

Improved selectivity of ZnNa3DTPA vs. Na5DTPA to abstract Cd2+ from plasma proteins in vitro.

Since no chelating agent has been clinically approved for the treatment of Cd(2+) intoxicated humans, we have previously investigated diethylenetriaminepentaacetic acid (Na5DTPA) to abstract this toxic metal from plasma proteins in vitro. In addition to the complete mobilization of Cd(2+), an inadvertent abstraction of Zn(2+) from metalloproteins was also observed. Therefore, we have evaluated the effect of ZnNa3DTPA on the plasma distribution of Ca, Cd, Cu, Fe and Zn after its addition to Cd(2+)-spiked rabbit plasma. ZnNa3DTPA was as efficient as Na5DTPA in abstracting Cd(2+) (100% removal), but it did not abstract Zn(2+). Thus, complexes between essential metals (e.g. Zn(2+)) and known chelating agents emerge as feasible candidates to decrease potential adverse side effects in patients.

Liquid chromatography-inductively coupled plasma-based metallomic approaches to probe health-relevant interactions between xenobiotics and mammalian organisms.

In mammals, the transport of essential elements from the gastrointestinal tract to organs is orchestrated by biochemical mechanisms which have evolved over millions of years. The subsequent organ-based assembly of sufficient amounts of metalloproteins is a prerequisite to maintain mammalian health and well-being. The chronic exposure of various human populations to environmentally abundant toxic metals/metalloid compounds and/or the deliberate administration of medicinal drugs, however, can adversely affect these processes which may eventually result in disease. A better understanding of the perturbation of these processes has the potential to advance human health, but their visualization poses a major problem. Nonetheless, liquid chromatography-inductively coupled plasma-based 'metallomics' methods, however, can provide much needed insight. Size-exclusion chromatography-inductively coupled plasma atomic emission spectrometry, for example, can be used to visualize changes that toxic metals/medicinal drugs exert at the metalloprotein level when they are added to plasma in vitro. In addition, size-exclusion chromatography-inductively coupled plasma mass spectrometry can be employed to analyze organs from toxic metal/medicinal drug-exposed organisms for metalloproteins to gain insight into the biochemical changes that are associated with their acute or chronic toxicity. The execution of such studies-from the selection of an appropriate model organism to the generation of accurate analytical data-is littered with potential pitfalls that may result in artifacts. Drawing on recent lessons that were learned by two research groups, this tutorial review is intended to provide relevant information with regard to the experimental design and the practical application of these aforementioned metallomics tools in applied health research.

Probing bioinorganic chemistry processes in the bloodstream to gain new insights into the origin of human diseases.

In the context of elucidating the origin of human diseases, past poisoning epidemics have revealed that exceedingly small doses of inorganic environmental pollutants can result in dramatic effects on human health. Today, numerous organic and inorganic pollutants have been quantified in human blood, but the interpretation of these concentrations remains--from a public health point of view--problematic. Conversely, the biomolecular origin for several grievous human diseases is essentially unknown. Taken together and viewed in the context of recent bioinorganic research findings, the established human blood concentrations of toxic metals and metalloids may be functionally connected with the etiology of specific human diseases. To unravel the underlying biomolecular mechanisms, and taking into account the basic flow of dietary matter through mammalian organisms, a better understanding of the bioinorganic chemistry of toxic metals and metalloid compounds in the bloodstream is emerging as a promising avenue for future research. To this end, the concerted application of modern proteomic methodologies, synchrotron-based X-ray absorption spectroscopy and established spectroscopic techniques will contribute to better define the role that blood-based bioinorganic chemistry-related processes play in the origin of human diseases. The application of this and other modern proteomic methodologies could contribute to a better understanding of the role that blood-based bioinorganic chemistry-related processes play in the origin and etiology of human diseases.

Remarkable effect of mobile phase buffer on the SEC-ICP-AES derived Cu, Fe and Zn-metalloproteome pattern of rabbit blood plasma.

The development of an analytical method to quantify the major Cu, Fe and Zn-containing metalloproteins in mammalian plasma has been recently reported. This method is based on the separation of plasma proteins by size exclusion chromatography (SEC) followed by the on-line detection of the metalloproteins by an inductively coupled plasma atomic emission spectrometer (ICP-AES). To assess whether the mobile phase buffer can affect the SEC-ICP-AES-derived metalloproteome pattern, thawed rabbit plasma was analyzed using phosphate buffered saline (PBS)-buffer (0.15 M, pH 7.4), Tris-buffer (0.1 and 0.05 M, pH 7.4), Hepes-buffer (0.1 M, pH 7.4) or Mops-buffer (0.1 M, pH 7.4). In contrast to the Cu-specific chromatograms, the Fe and Zn-specific chromatograms that were obtained with Tris, Hepes and Mops-buffer were considerably different from those attained with PBS-buffer. The Tris, Hepes and Mops-buffer mediated redistribution of ~25% plasma Zn(2+) from <100 kDa to >100-600 kDa plasma proteins and to a smaller extent to a <10 kDa (Tris)(2)Zn(2+)-complex can be rationalized in terms of the abstraction of Zn(2+) from the weak binding site on albumin. In contrast, only Hepes and Mops-buffer redistributed ~20% of plasma Fe(3+) from the <100 kDa to the >600 kDa elution range. Based on these results and considering that the utilization of PBS-buffer has previously resulted in the detection of a number of Cu, Fe and Zn-containing metalloentities in rabbit plasma that was most consistent with literature data, this mobile phase buffer is recommended for metallomic studies regarding mammalian blood plasma.