Non-invasive Sampling Techniques Research Articles

Detection of Filariid Infections in Mexican Primate Populations Through qPCR.

Filariae are parasitic nematodes of high veterinary and medical importance, responsible for some acute tropical diseases. They are transmitted through the bite of hematophagous vectors such as biting midges and blackflies. Filariae are among the most prevalent vector-borne parasitoses in Neotropical primates in which severe infections can cause inflammatory reactions and tissue damage. Given the location inside the host (peritoneal cavity, bloodstream, and lymphatics), the detection of filariid nematodes is challenging and is mostly postmortem; hence the scarcity of studies on the prevalence of filariae in wild primate populations. Here, we report the prevalence of filariid infections in free-ranging populations of Geoffroy's spider (Ateles geoffroyi) and black howler (Alouatta pigra) monkeys across southern Mexico, using a combination of noninvasive sampling and molecular diagnostic techniques. Fecal samples were screened for filariid DNA by qPCR protocols. A total of 88 samples were examined with an overall prevalence of 26%. Filariae were slightly more common in spider monkeys compared to howler monkeys. This study constitutes the first report of the prevalence of infection of filariid nematodes in populations of wild spider monkey across southern Mexico, and the first reporting of filariae in black howler monkeys, as part of a new era of primate parasitology and the diagnostics of parasite infections in light of the everyday more affordable molecular tools.

Use of long-term underwater camera surveillance to assess the effects of the largest Amazonian hydroelectric dam on fish communities

The increase in the construction of mega dams in tropical basins is considered a threat to freshwater fish diversity. Although difficult to detect in conventional monitoring programs, rheophilic species and those reliant on shallow habitats comprise a large proportion of fish diversity in tropical basins and are among the most sensitive species to hydropower impacts. We used Baited Remote Underwater Video (BRUV), an innovative, non-invasive sampling technique, to record the impacts caused by Belo Monte, the third largest hydropower project in the world, on fishes inhabiting fast waters in the Xingu River. BRUV were set in a river stretch of ~ 240 km for 7 years, 2 before and 5 after the Belo Monte operation. We explored the spatial and temporal variation in fish diversity (α, β, and γ) and abundance (MaxN) using generalized additive models. We also investigated the variation of environmental variables and tested how much information we gained by including them in the diversity and abundance models. Belo Monte altered the flow regime, water characteristics, and fishery yield in the Xingu, resulting in changes in the fish community structure. Temporally, we observed sharp declines in α diversity and abundance, far exceeding those from a previous study conducted with more conventional sampling methods (i.e., catch-based) in the region. γ-diversity was also significantly reduced, but we observed a non-expected increase in β diversity over time. The latter may be associated with a reduction in river connectivity and an increase in environmental heterogeneity among river sectors. Unexpected signs of recovery in diversity metrics were observed in the last years of monitoring, which may be associated with the maintenance of flow levels higher than those previously planned. These results showed that BRUV can be a useful and sensitive tool to monitor the impacts of dams and other enterprises on fish fauna from clear-water rivers. Moreover, this study enhances our comprehension of the temporal variations in freshwater fish diversity metrics and discusses the prevalent assumption that a linear continuum in fish-structure damage associated with dam impoundments may exhibit temporal non-linearity.

A Survey for Atypical Brucella Species in Wild-Caught Marine Toads (Rhinella marina) Using Qualitative PCR: A Pilot Study

Abstract The genus Brucella is a group of intracellular bacterial pathogens documented to cause brucellosis, a zoonotic disease of worldwide concern. In the past decade, reported cases of atypical Brucella species in amphibians have increased. These species are considered atypical based on their tropism, phenotypic characteristics, and phylogeny differences. Atypical Brucella species can be misdiagnosed due to their similarity to classical Brucella species and closely related genera. Recent literature has provided evidence for the zoonotic potential of atypical Brucella linked to amphibians. Therefore, understanding the prevalence of these atypical Brucella species in captive amphibian populations is significant for amphibian health, conservation, and public health. This pilot study evaluated the prevalence of atypical Brucella species in a population of marine toads (Rhinella marina) using a validated pan-Brucella qualitative PCR. Samples were analyzed from 56 asymptomatic wild-caught marine toads from Miami, FL, USA, using skin swab samples. Brucella DNA was not detected in any of the skin swab samples analyzed. This pilot study represents a viable non-invasive sampling technique and PCR assay for rapidly screening populations of amphibians for the presence of atypical Brucella species.

Evaluation of Non-Invasive Sampling Techniques for the Molecular Surveillance of Equid Herpesviruses in Yearling Horses.

Equid alphaherpesvirus 1 (EHV-1) is a highly contagious respiratory tract pathogen of horses, and infection may be followed by myeloencephalopathy or abortion. Surveillance and early detection have focused on PCR assays using less tolerated nasal swabs. Here, we assess non-invasive non-contact sampling techniques as surveillance tools in naturally equid gammaherpesvirus 2-shedding horses as surrogates for EHV-1. Horses were individually housed for 10 h periods on 2 consecutive days. Sampling included nasal swabs, nostril wipes, environmental swabs, droplet-catching devices, and air sampling. The latter was completed via two strategies: a combined air sample collected while going from horse to horse and a collective air sample collected at a stationary central point for 6 h. Samples were screened through quantitative PCR and digital PCR. Nine horses on day 1 and 11 horses on day 2 were positive for EHV-1; overall, 90.9% of the nostril wipes, 81.8% of the environmental surfaces, and 90.9% of the droplet-catching devices were found to be positive. Quantitative analysis showed that the mean DNA copies detection per cm2 of nostril wipe sampled concentration (4.3 × 105 per day) was significantly (p < 0.05) comparable to that of nasal swabs (3.6 × 105 per day) followed by environmental swabs (4.3 × 105 per day) and droplet catchers (3.5 × 103 per day), respectively. Overall, 100% of the air samples collected were positive on both qPCR and dPCR. In individual air samples, a mean concentration of 1.0 × 104 copies of DNA were detected in per m3 air sampled per day, while in the collective air samples, the mean concentration was 1.1 × 103. Environmental samples look promising in replacing direct contact sampling. Environmental and air sampling could become efficient surveillance tools at equestrian events; however, it needs threshold calculations for minimum detection levels.

Can hair snares provide a reliable method for estimating abundance of an exotic ungulate?

Mitigating exotic species invasions requires reliable information on abundance and source-sink dynamics to evaluate effectiveness of control efforts. Despite being present in the southeastern United States for centuries, wild pigs (Sus scrofa) continue to invade new areas, including the Sam D. Hamilton Noxubee National Wildlife Refuge (NNWR) in Mississippi, a forested landscape with an emerging wild pig population. Considering the early stage of invasion, we designed and implemented a non-invasive hair sampling technique to estimate abundance and relatedness of wild pigs within the NNWR. Following 8-week sampling periods in 2021 (n = 160 samples) and 2022 (n = 159 samples), we identified 38 and 26 unique individuals, respectively, although recapture rates were too low to yield precise abundance estimates. We also identified low heterozygosity indicating high relatedness among our population which likely reflected lack of barrier features to gene flow and/or a limited number of external source populations. Finally, we observed positive trends in viable samples collected relative to sampling week, suggesting more optimal sampling period(s) may exist (e.g., autumn). We also acknowledge that use of grain in more accessible areas rather than our remote approach using scent-only attractants may increase hair snare attractiveness and wild pig encounters, while permitting increased sampling frequency. Collectively, these modifications (i.e., altered season, bait type, and access) provide viable pathways to increase precision for abundance estimates. When used concomitantly with ongoing monitoring and control efforts, non-invasive hair sampling will contribute to a more holistic understanding of this exotic species and better inform management actions.

Pregnancy maintenance and fetal loss assessment in Holstein cows through analyzing pregnancy-associated glycoproteins in milk

Fetal loss (FL) from the 45th day of gestation until calving can impose a significant economic burden on dairy farmers, resulting in lost profits and increased production costs. Pregnancy-associated glycoprotein (PAG) is commonly used for detecting pregnancy in cows. PAG is secreted by binucleated trophoblast cells of the placenta and regulated by more than 24 genes. The purpose of this study was to determine the PAG threshold for FL and a probability of pregnancy maintenance until calving based on milk PAG levels. Our results reveal that primiparous and multiparous cows that maintained pregnancy until the 40th week exhibited higher PAG sample-negative (SN) values in their milk in the 6th week of gestation than did those that experienced FL later in gestation. Pregnant cows with higher PAG SN values in the 6th week of gestation were more likely to maintain their pregnancies. The area under the receiver operating characteristic curve for predicting the probability of pregnancy maintenance was 0.722 for our prediction model. On the other hand, a milk PAG SN value of <0.192 indicated 95 % confidence that FL would occur between the 7th and 40th weeks of gestation. Milk PAG testing is a noninvasive sampling technique that does not induce additional stress in lactating cows. The study reveals that PAG SN values increase significantly in Holstein cows during the 6th week of gestation. The predictive model developed was effective in forecasting pregnancy outcomes up to the 40th week of gestation or calving. The model's performance is moderately good for field application and could be a useful tool for dairy producers.

Analysis of winter survival strategies of sympatric black-necked cranes, and common cranes from the perspective of diet and gut microbiota

Wintering migratory birds have to travel thousands of kilometers between their breeding and wintering grounds because of their distinct habits. They experience complex and changing survival pressures throughout the winter. This study is the first to investigate the overwintering strategies of sympatric foraging black-necked cranes and common cranes from the perspectives of food and gut microbiota. Non-invasive sampling technique was used to collect stool samples. Sanger sequencing was used to determine the origin of the crane species identified from the samples. The analysis of plant-based food, the gut microbiota, and their interactions in the faeces was performed using high-throughput sequencing technology. It was found that in environments with limited aquatic vegetation, both the black-necked crane and the common crane increased their consumption of potatoes, accounting for more than 50% of their diets. Black-necked cranes also competed with common cranes for food, and they quickly adapted to changes in diet by horizontally transmitting their gut microbiota. To meet body needs, the common cranes—which could not compete with the black-necked cranes—constantly searched for new food sources. They increase the metabolism of amino acids, lipids and these metabolites to adapt to the competitive pressures and dietary changes. This study advances our knowledge of how diet and gut microbiota interact in plateau wetland birds, examining the competitive tactics of interwintering black-necked and common cranes', and provides recommendations for the re-vegetation in the Caohai wetlands and artificial feeding of the two crane species.

A Review of the Occurrence of Metals and Xenobiotics in European Hedgehogs (Erinaceus europaeus).

Monitoring data from several European countries indicate that European hedgehog (Erinaceus europaeus) populations are declining, and research exploring the causes of the decline, including exposure to potentially harmful xenobiotics and metals, may inform conservation initiatives to protect this species in the wild. Hedgehogs are ground-dwelling mammals, feeding on a range of insects, slugs, snails, and earthworms, as well as eggs, live vertebrates, and carrion, including carcasses of apex predator species representing higher levels of the food chain. Consequently, hedgehogs come into close contact with contaminants present in their habitats and prey. This review investigated the studies available on the subject of the occurrence of metals and organic xenobiotics in hedgehogs. This study found that a vast range of different pesticides; persistent organic pollutants (POPs), including organochlorine compounds and brominated flame retardants (BFRs); as well as toxic heavy metals could be detected. Some compounds occurred in lethal concentrations, and some were associated with a potential adverse effect on hedgehog health and survival. Due to their ecology, combined with the opportunity to apply non-invasive sampling techniques using spines as sampling material, we suggest that the European hedgehog is a relevant bioindicator species for monitoring the exposure of terrestrial wildlife to potential toxicants in urban and rural environments.

Echolocation call characterization of insectivorous bats from caves and karst areas in southern Luzon Island, Philippines

Bats are excellent bioindicators and are increasingly used to assess ecosystem health and monitor changes in the environment. Due to increased awareness of the potential transmission of pathogens from bats to humans and recognizing the limitations of traditional bat sampling methods, the use of of non-invasive sampling techniques such as bat recorders were recommended for field-based monitoring studies. In the Philippines, however, bat bioacoustics is still a growing field, and the scarcity of acoustic data hinders the use of echolocation calls to conduct accurate inventories and population monitoring of echolocating bats. Here, we recorded and characterized echolocation calls of insectivorous bats from caves and karst areas located in southern Luzon Island, Philippines. In addition, we compared our results with other studies performed within and outside the country to identify possible regional and local variation in acoustic characters for some species. A total of 441 echolocation calls were recorded from six bat families: Hipposideridae (five species), Rhinolophidae (five species), Vespertilionidae (three species), Miniopteridae (two species), Megadermatidae (one species), and Emballonuridae (one species). Discriminant function analyses (DFA) with leave-one-out cross validation correctly classified bats emitting calls dominated with a constant frequency (CF) component (rhinolophids and hipposiderids) with &gt;97% success and those producing frequency modulated (FM) calls (Miniopteridae and Vespertilionidae) with 88.9% success. We report echolocation calls for Philippine population of two species (Megaderma spasma and Hipposideros lekaguli) for the first time. Moreover, we present geographical variations in call frequencies for some species by comparing previously reported acoustic data elsewhere across the species’ range. This underscores the importance of establishing a readily accessible and comprehensive local reference library of echolocation calls which would serve as a valuable resource for examining taxonomic identities of echolocating bats, particularly those whose calls exhibit biogeographic variations.

Prediction of Potassium (K) Content in Soil Analysis Utilizing Near-Infrared (NIR) Spectroscopy

The relationship between NIR spectroscopy spectral absorbance and potassium (K) is studied as a representative factor for analysing soil's nutritional content. To determine the potassium content of soil samples without resorting to intrusive and time-consuming chemical analysis techniques, NIR spectroscopy sampling techniques have been tested. Spectrum absorption data from 900 nm to 1600 nm were discovered to correspond with potassium values and were then analysed using a number of pre-processing procedures. Five techniques, Multiplicative Scatter Correction (MSC), Multiplicative Scatter Correction using Common Amplification (MSSCA), Multiplicative Scatter Correction using Common Offset (MSCCO), Detrending (DT), and Mean Normalization (MN), have been identified as the most effective. Using the Partial Least Squares Regression (PLSR) model, both calibration and prediction data are evaluated. In the end study, the MSCCA method was determined to be the most effective pre-processing method for both calibration and prediction outcomes, with R2 values of 0.9998 for calibration and RMSE values of 0.0600 for prediction. Utilising PLSR model and the MSCCA preprocessing method, the relationship between NIRS absorbance data and potassium may be determined. Consequently, we may infer that the NIRS approach can be utilised to detect amount of potassium in soil analysis employing a less time-consuming, non-invasive, and labour-intensive sampling technique.

Inflammatory biomarkers in sebum for identifying skin damage in patients with a Stage I pressure ulcer in the pelvic region: A single centre observational, longitudinal cohort study with elderly patients.

Pressure Ulcers (PU) are a major burden for affected patients and healthcare providers. Current detection methods involve visual assessments of the skin by healthcare professionals. This has been shown to be subjective and unreliable, with challenges associated with identifying erythema in darker colour skin. Although there exists a number of promising non-invasive biophysical techniques such as ultrasound, capacitance measurements, and thermography, the present study focuses on directly measuring the changes in the inflammatory status of the skin and underlying tissues. Therefore, in this study, we aim to analyse inflammatory cytokines collected through non-invasive sampling techniques to detect early signs of skin damage. Thirty hospitalised patients presenting with Stage I PU were recruited to evaluate the inflammatory response of skin at the site of damage and an adjacent healthy control site. Sebutapes were collected over three sessions to investigate the temporal changes in the inflammatory response. The panel of cytokines investigated included high-abundance cytokines, namely, IL-1α and IL-1RA, and low abundance cytokines; IL-6, IL-8, TNF-α, INF-γ, IL-33, IL-1β and G-CSF. Spatial and temporal differences between sites were assessed and thresholds were used to determine the sensitivity and specificity of each biomarker. Theresultssuggest significant (P < .05) spatial changes in the inflammatory response, with upregulation of IL-1α, IL-8, and G-CSF as well as down-regulation of IL-1RA over the Stage I PU compared with the adjacent control site. There were no significant temporal differences between the three sessions. Selected cytokines, namely, IL-1α, IL-1RA, IL-8, G-CSF, and the ratio IL-1α/IL-1RA offered clear delineation in the classification of healthy and Stage-I PU skin sites, with receiver operating characteristic curves demonstrating high sensitivity and specificity. There were limited influences of intrinsic and extrinsic factors on the biomarker response. Inflammatory markers provided a high level of discrimination between the sites presenting with Stage I PU and an adjacent healthy skin site, in a cohort of elderly inpatients. Indeed, the ratio of IL-1α to IL-1RA provided the highest sensitivity and specificity, indicative that inflammatory homeostasis is affected at the PU site. There was a marginal influence of intrinsic and extrinsic factors, demonstrating the localised effects of the inflammation. Further studies are required to investigate the potential of inflammatory cytokines incorporated within Point of Care technologies, to support routine clinical use.

Biomarkers and their potential for detecting livestock plant poisonings in Western North America.

The United States National Cancer Institute defines a biomarker as: "A biological molecule found in blood, other body fluids, or tissues that is a sign of a normal or abnormal process, or of a condition or disease." In Veterinary Medicine, biomarkers associated with plant poisonings of livestock have great utility. Since grazing livestock poisoned by toxic plants are often found dead, biomarkers of plant poisoning allow for a more rapid postmortem diagnosis and response to prevent further deaths. The presence and concentration of toxins in poisonous plants are biomarkers of risk for livestock poisoning that can be measured by the chemical analysis of plant material. More difficult is, the detection of plant toxins or biomarkers in biological samples from intoxicated or deceased animals. The purpose of this article is to review potential biomarkers of plant poisoning in grazing livestock in the Western North America including recently investigated non-invasive sampling techniques. Plants discussed include larkspur, lupine, water hemlock, swainsonine-containing plants, selenium-containing plants, and pyrrolizidine alkaloid containing plants. Other factors such as animal age and sex that affect plant biomarker concentrations in vivo are also discussed.

Large-scale grid-based detection in occupancy surveys of a threatened small mammal: A comparison of two non-invasive methods

Monitoring the status and trends of wildlife is key to understand how species respond to natural and human-derived threats, and to evaluate and improve conservation planning and management. Large-scale, grid-based assessment of species distribution, abundance, and population trends over time is an important component of biodiversity monitoring. However, such assessments still present important challenges related, for instance, to how the choice of the sampling method may affect species detectability and thus, overall data accuracy. Here, we address this issue, focusing on the Cabrera vole (Microtus cabrerae), a threatened small mammal, listed in the Habitats Directive (Annexes II and IV), hence requiring regular evaluation of its population status and trends. We used occupancy modelling to estimate method-specific detection probability of the species over large-scale, grid-based (10 × 10 km2) surveys relying on two non-invasive sampling techniques: sign surveys and owl pellet analysis. Results provided evidence for a greater cost-effectiveness of sign surveys compared to owl pellet analysis for detecting the species in occupancy surveys, suggesting that large-scale population monitoring of Cabrera voles (or other species also producing easily identifiable signs of their presence) may fairly rely on sign-surveys. Overall, our study supported the view that while owl pellet analysis provides a valuable option when the aim is to assess small mammal assemblages (i.e. multiple species) in a region, other complementary methods may be required to increase the detection probability of certain species that because of their secretive behaviour or rarity remain less predated by owls. We thus argue that the choice of the sampling method should be context-dependent and evaluated based on the study aims, the surveyed area (i.e. local factors), the target species (i.e. life history traits) and the available resources. Based on our results we recommend that researchers and managers explore survey-design trade-offs to ensure the proposed designs have sufficient power to detect real population trends.

Cuticular Swabs and eDNA as Non-Invasive Sampling Techniques to Monitor Aphanomyces astaci in Endangered White-Clawed Crayfish (Austropotamobius pallipes Complex)

In endangered crayfish conservation projects, it is paramount to map the distribution of the causative agent of crayfish plague, Aphanomyces astaci, in native populations. Considering the inapplicability of the destructive cuticular sampling protocol for monitoring endangered populations, we explored the use of non-invasive sampling techniques to detect this pathogen with molecular assays. In the present study, we exploited environmental DNA (testing increasing water volumes combined with different filter porosities) and cuticular swabs to collect A. astaci DNA. In addition, we evaluated the impact of the storage method on DNA preservation during field activities. After the first evaluations performed on both highly infected Austropotamobius pallipes and carrier Procambarus clarkii specimens in laboratory conditions, these sampling techniques were applied to wild populations of white-clawed crayfish. Our findings highlight better results with the filtration of 5 L of water with filters of 2.7 µm porosity for eDNA analysis and demonstrate that cuticular swabbing is equally effective as the World Organisation of Animal Health’s protocol. Storage in absolute ethanol proved to be the best solution to preserve swabs and filter samples for up to a week at room temperature. In conclusion, we suggest an integration of both sampling methods when monitoring A. astaci for conservation purposes.

Individual Identification of Panthera pardus saxicolor Using Non-Invasive Sampling and Molecular Techniques in Iran: A Case Study in Parvar Protected Area

The Persian leopard (Panthera pardus saxicolor) is an endangered species widely distributed across Iran. Rough estimates indicate 550–850 Persian leopard individuals inhabit the country, which comprises more than two thirds of its global population. Population monitoring of this large carnivore in Iran is one of the main objectives of current conservation planning. Therefore, adopting a more diversified methodology to achieve reliable, cost-effective, and pragmatic measures is urgently needed. We conducted a study for individual identification of the Persian leopard in Parvar Protected Area via fecal sampling and molecular tools and attempted to test the feasibility of this approach. We used 12 previously reported polymorphic microsatellite loci, of which only five were qualified for genotyping analysis. Finally, nine leopard individuals were identified. We measured the ability of the five loci in distinguishing individuals by P(ID)sib. The cumulative observed probability of identity and probability of identity for sibling individuals were estimated to be 0.005 and 0.05, respectively. Data presented on spatial distribution of leopards in this study could help better understand the behavioral ecology and conservation biology of the species. Moreover, our findings will assist future research in developing methodologies for large-scale studies and providing data for effective wildlife conservation.

Effects of season and diel cycle on hydroacoustic estimates of density, Target Strength, and vertical distribution of fish in Yudong Reservoir, a plateau deep water reservoir in southwest China in a plateau deep

Hydroacoustics is a non-invasive fish stock assessment sampling technique that plays an important role in fishery science and management. However, non-standard hydroacoustic surveys could lead to biased results, and the factor of the sampling period (e.g., season and diel cycle) is extremely critical as it can greatly affect hydroacoustic results. Efforts to improve the accuracy and credibility of the hydroacoustic survey results are getting more and more attention. Thus, we conducted two diel hydroacoustic surveys in situ in summer and winter to detect whether there were diel and seasonal differences in density, Target Strength (TS) and vertical distribution of fish. The results indicated that nighttime had significantly higher fish mean density than daytime in summer and winter. No significant difference between summer and winter daytime, however, significant difference between summer and winter nighttime, but this bias could be accepted from the fisheries management perspective; The mean TS of the summer daytime was significantly higher than that of summer nighttime, winter daytime and winter nighttime, but there were no significant differences among summer nighttime, winter daytime, and winter nighttime, mainly due to mean TS may be overestimated from fish schooling behavior during summer daytime; The fish vertical distribution had significant seasonal correlations and was more dispersed in different water layers during the nighttime, proving that the assessment was better at nighttime than during the daytime. Consequently, the hydroacoustic surveys in Yudong Reservoir and other similar plateau deep water reservoirs should be performed at nighttime, which will yield relatively accurate density and TS, and dispersed vertical distribution of fish.

Do Pharmaceuticals in the Environment Pose a Risk to Wildlife?

The vast majority of knowledge related to the question "To what extent do pharmaceuticals in the environment pose a risk to wildlife?" stems from the Asian vulture crisis (>99% decline of some species of Old World vultures on the Indian subcontinent related to the veterinary use of the nonsteroidal anti-inflammatory drug [NSAID] diclofenac). The hazard of diclofenac and other NSAIDs (carprofen, flunixin, ketoprofen, nimesulide, phenylbutazone) to vultures and other avian species has since been demonstrated; indeed, only meloxicam and tolfenamic acid have been found to be vulture-safe. Since diclofenac was approved for veterinary use in Spain and Italy in 2013 (home to ~95% of vultures in Europe), the risk of NSAIDs to vultures in these countries has become one of the principal concerns related to pharmaceuticals and wildlife. Many of the other bodies of work on pharmaceutical exposure, hazard and risk to wildlife also relate to adverse effects in birds (e.g., poisoning of scavenging birds in North America and Europe from animal carcasses containing pentobarbital, secondary and even tertiary poisoning of birds exposed to pesticides used in veterinary medicine as cattle dips, migratory birds as a vector for the transfer of antimicrobial and antifungal resistance). Although there is some research related to endocrine disruption in reptiles and potential exposure of aerial insectivores, there remain numerous knowledge gaps for risk posed by pharmaceuticals to amphibians, reptiles, and mammals. Developing noninvasive sampling techniques and new approach methodologies (e.g., genomic, in vitro, in silico, in ovo) is important if we are to bridge the current knowledge gaps without extensive vertebrate testing. Environ Toxicol Chem 2024;43:595-610. © 2022 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

Associations between hair trace mineral concentrations and the occurrence of treponeme-associated hoof disease in elk (Cervus canadensis)

BackgroundTrace minerals are important for animal health. Mineral deficiency or excess can negatively affect immune function, wound healing, and hoof health in domestic livestock, but normal concentrations and health impairment associated with mineral imbalances in wild animals are poorly understood. Treponeme-associated hoof disease (TAHD) is an emerging disease of free-ranging elk (Cervus canadensis) in the U.S. Pacific Northwest. Selenium and copper levels identified in a small number of elk from areas where TAHD is established (i.e., southwestern Washington) suggested a mineral deficiency may have increased susceptibility to TAHD. Our objectives were to determine trace mineral concentrations using hair from elk originating in TAHD affected areas of Washington, California, Idaho, and Oregon and assess their associations with the occurrence of the disease.ResultsWe identified limited associations between TAHD occurrence and severity with hair mineral concentrations in 72 free-ranging elk, using Firth’s logistic regression and multinomial regression models. We found consistent support for a priori hypotheses that selenium concentration, an important mineral for hoof health, is inversely associated with the occurrence of TAHD. Less consistent support was observed for effects of other minerals previously associated with hoof health (e.g., copper or zinc) or increased disease risk from potential toxicants.ConclusionTrace mineral analysis of hair is a non-invasive sampling technique that offers feasibility in storage and collection from live animals and carcasses. For some minerals, levels in hair correlate with visceral organs that are challenging to obtain. Our study using hair collected opportunistically from elk feet submitted for diagnostic investigations provides a modest reference of hair mineral levels in elk from the U.S. Pacific Northwest that may be useful in future determination of reference ranges. Although our results revealed high variability in mineral concentrations between elk, consistent relationship of possibly low selenium levels and TAHD suggest that further investigations are warranted.

First report of a severe nasopulmonary acariasis caused by Orthohalarachne diminuata Doetschman, 1944 (Acari: Halarachnidae) in a captive South American sea lion (Otaria flavescens Shaw, 1800).

Obligatory endoparasitic mites of the genera Halarachne Allman, 1847 and Orthohalarachne Newell, 1947 (Acari: Halarachnidae) parasitize different segments of the respiratory tract of marine mammals, including pinnipeds and sea otters, and infestations can cause asymptomatic to serious respiratory diseases. However, knowledge on biology, pathogenic potential and occurrence of halarachnid mites infesting pinnipeds, especially in captivity, is scarce. A two-year-old South American sea lion (Otaria flavescens Shaw, 1800) male, born and held at the Vienna Zoo, was anesthesized for routine pre-transport examinations, including computed tomography, bronchoalveolar lavage, and blood sampling. During the final phase of general anesthesia, the individual abruptly became apneic and died despite all attempts at resuscitation. At necropsy, 45 highly motile whitish millimeter-sized structures were macroscopically detected in the trachea, bifurcatio tracheae and main bronchi and were identified as adult stages of Orthohalarachne diminuata Doetschman, 1944 following morphological descriptions. After trepanation of the nasal cavity and sinus paranasalis, a total of 407 larval and 3 nymphal specimens distributed in clusters were detected. Macroscopically, sinus mucosa showed hyperemia and multiple petechial hemorrhages. Histopathological analyses of paranasal sinuses revealed mite cross-sections surrounded by sanioserous exudate and epithelial exfoliation. For the first time, O. diminuata was molecularly characterized and phylogenetically analyzed based on its 16S rDNA. Our study constitutes the first record of a severe O. diminuata infestation in captive O. flavescens and one of the few host-parasite records in general. We present clinical data and pathological results, the first scanning electron microscopic images of a O. diminuata larval stage and discuss the etiology of this autochthonous infestation, possible transmission pathways and detrimental effects. Further studies on biology and pathogenic effects of halarachnid mites, as well as on the development of non-invasive sampling techniques are essentially required for a better understanding of (ortho-)halarachnosis in pinnipeds held in zoological gardens.

Salivary molecules as indicators of hydration status

Introduction: Maintaining an appropriate level of hydration is essential to health, especially when doing physical exercise. Thus, the importance of monitoring the before, during and after exercise status. Therefore, knowing and studying the changes and indicators that may provide reliable information on fluctuations in a simple, quick manner, via non-invasive sampling techniques becomes essential. Material and Methods: The data analyzed and processed (α-amylase, K+, Cl-, cortisol, total protein, and osmolality concentration in saliva) in this study are available on the free access research platform for complex physiological signals (https://physionet.org/content/qde/1.0.0/). Ten male subjects voluntarily took part in this study. The median and standard deviation of height, body weight, and age were 179 ±7.5 cm. 79.3 ±9.0 kg. and 25.5 ±3.7 years, respectively. Results: The α-amylase, K+, cortisol, total protein, and osmolality concentration in saliva showed significant variations for moderate dehydration levels. For the cases of Cl- concentration changes in saliva, statistically significant changes can be observed at a much earlier stage (in mild dehydration conditions). Conclusions: The results suggest that Cl- concentration in saliva might be used as an indicator for the hydration status. Therefore, a scale to monitor the hydration status is proposed.