Non-invasive Sampling Techniques Research Articles

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.

Mitochondrial genetic diversity of the Greater Cane rat (Thryonomys swinderianus) populations from the Eastern Arc Mountains ecosystem, Tanzania.

Management of most herbivorous small mammal species considered to be pests in Africa is still challenging partly because of the paucity of information on their biological traits that would help to manage their destructive impacts. This gap also precludes the potential for tapping species with potential food-value to improving the economy of rural communities through, for example, sustainable game farming programs in Africa. This study investigates the genetic diversity and population demography of the African Greater Cane rat (AGC), a rodent pest of crops and game species inhabiting two isolated blocks of the Eastern Arc Mountains (EAMs), Tanzania to contribute to the species management and conservation. We used non-invasive sampling techniques and DNA sequencing of the D-loop region of MtDNA (515bp) from 46 cane rats (Thryonomys swinderianus) samples to characterize the genetic diversity and structure of the species and potential population threats faced in natural habitats. We found 25 haplotypes:15 from Uluguru and 9 from Udzungwa mountains populations, containing 49 polymorphic regions (32 parsimoniously informative and 17 singleton sites). Haplotype diversity (range: 0.849-0.995) did not differ substantially across populations but the median haplotype diversity for Udzungwa South was overall lower than for other populations. Nucleotide diversity averaged 0.00641, 0.01528, 0.0111 and 0.01313, respectively for Udzungwa South, Udzungwa North, Uluguru Rural and Uluguru Urban, suggesting high genetic diversity within the four populations. Analysis of molecular variance (AMOVA) indicated significantly high genetic differences between the four populations (FST = 0.16, p = 0.00098) whereas neutrality test (FU's Fs) values were negative, indicating historical population expansion. Similarly, the Bayesian skyline analysis indicated a recent demographic expansion suggesting limited bottlenecks in the recent past in this population. Our results show the AGC population in EAMs consists of four distinct populations which have experienced a recent population expansion, especially among the urban population due perhaps to influence of urbanization process that may have favored assisted species movements across the rural-urban landscapes. Future research should focus on understanding impact of geographical isolation on the genetic structure and diversity of this species.

An Extensive Study of Phenol Red Thread as a Novel Non-Invasive Tear Sampling Technique for Proteomics Studies: Comparison with Two Commonly Used Methods.

Tear samples are considered in recent publications as easily, noninvasively collectible information sources for precision medicine. Their complex composition may aid the identification of biomarkers and the monitoring of the effectiveness of treatments for the eye and systemic diseases. Sample collection and processing are key steps in any analytical method, especially if subtle personal differences need to be detected. In this work, we evaluate the usability of a novel sample collection technique for human tear samples using phenol red threads (cotton thread treated with the pH indicator phenol red), which are efficiently used to measure tear volume in clinical diagnosis. The low invasiveness and low discomfort to the patients have already been demonstrated, but their applicability for proteomic sample collection has not yet been compared to other methods. We have shown, using various statistical approaches, the qualitative and quantitative differences in proteomic samples collected with this novel and two traditional methods using either glass capillaries or Schirmer’s paper strips. In all parameters studied, the phenol red threads proved to be equally or even more suitable than traditional methods. Based on detectability using different sampling methods, we have classified proteins in tear samples.

An extensive study of phenol red thread as a novel non-invasive tear sampling technique for proteomics studies: comparison with two commonly used methods

An extensive study of phenol red thread as a novel non-invasive tear sampling technique for proteomics studies: comparison with two commonly used methods

Profiling the Atopic Dermatitis Epidermal Transcriptome by Tape Stripping and BRB-seq.

Tape stripping is a non-invasive skin sampling technique, which has recently gained use for the study of the transcriptome of atopic dermatitis (AD), a common inflammatory skin disorder characterized by a defective epidermal barrier and perturbated immune response. Here, we performed BRB-seq—a low cost, multiplex-based, transcriptomic profiling technique—on tape-stripped skin from 30 AD patients and 30 healthy controls to evaluate the methods’ ability to assess the epidermal AD transcriptome. An AD signature consisting of 91 differentially expressed genes, specific for skin barrier and inflammatory response, was identified. The gene expression in the outermost layers, stratum corneum and stratum granulosum, of the skin showed highest correlation between tape-stripped skin and matched full-thickness punch biopsies. However, we observed that low and highly variable transcript counts, probably due to low RNA yield and RNA degradation in the tape-stripped skin samples, were a limiting factor for epidermal transcriptome profiling as compared to punch biopsies. We conclude that deep BRB-seq of tape-stripped skin is needed to counteract large between-sample RNA yield variation and highly zero-inflated data in order to apply this protocol for population-wide screening of the epidermal transcriptome in inflammatory skin diseases.

Point of care diagnostics and non-invasive sampling strategy: a review on major advances in veterinary diagnostics

The use of point of care diagnostics (POCD) in animal diseases has steadily increased over the years since its introduction. Its potential application to diagnose infectious diseases in remote and resource limited settings have made it an ideal diagnostic in animal disease diagnosis and surveillance. The rapid increase in incidence of emerging infectious diseases requires urgent attention where POCD could be indispensable tools for immediate detection and early warning of a potential pathogen. The advantages of being rapid, easily affordable and the ability to diagnose an infectious disease on spot has driven an intense effort to refine and build on the existing technologies to generate advanced POCD with incremental improvements in analytical performance to diagnose a broad spectrum of animal diseases. The rural communities in developing countries are invariably affected by the burden of infectious animal diseases due to limited access to diagnostics and animal health personnel. Besides, the alarming trend of emerging and transboundary diseases with pathogen spill-overs at livestock-wildlife interfaces has been identified as a threat to the domestic population and wildlife conservation. Under such circumstances, POCD coupled with non-invasive sampling techniques could be successfully deployed at field level without the use of sophisticated laboratory infrastructures. This review illustrates the current and prospective POCD for existing and emerging animal diseases, the status of non-invasive sampling strategies for animal diseases, and the tremendous potential of POCD to uplift the status of global animal health care.