Digital Commons Network^™

Urine Proteomics Defines An Immune Checkpoint-Associated Nephritis Signature, James P Long, Shailbala Singh, Yanlan Dong, Cassian Yee, Jamie S Lin

Faculty, Staff and Student Publications

Immune checkpoint inhibitor (ICI) therapy is a cornerstone treatment for many cancers, but it can induce severe immunotoxicity, including acute interstitial nephritis (AIN). Currently, kidney biopsy is required to differentiate ICI-AIN from other causes of acute kidney injury (AKI). However, this invasive approach can lead to morbidity, delayed glucocorticoid treatment for patients with AIN, and unnecessarily prolonged suspension of ICI therapy in non-AIN patients. Delayed or incorrect diagnosis of ICI-AIN is particularly detrimental, as over 50% of patients are at risk of permanent renal damage. Thus, there is an urgent need for non-invasive biomarkers that can rapidly and accurately distinguish …

Review Of New Treatment Options For Toxoplasmosis, Kiley Roman

D.U.Quark

Toxoplasmosis is an infectious disease caused by the parasite, Toxoplasma Gondii. The parasite is often carried in cats and contaminated food and water. Toxoplasmosis is not serious most of the time but can be harmful to immunocompromised people, pregnant women, and their fetuses or babies. Treatments are commonly given to the immunocompromised. However, there are problems with the current treatments, in which they only treat active parasites and cannot remove the cysts created by disease progression. Due to the treatments’ inability to remove cysts, an infected person may not fully recover from the infection. This review evaluated the new …

Coronavirus Disease 2019 (Covid-19) Real World Data Infrastructure: A Big-Data Resource For Study Of The Impact Of Covid-19 In Patient Populations With Immunocompromising Conditions, James M. Crawford, Lynne Penberthy, Ligia A. Pinto, Keri N. Althoff, Magdalene M. Assimon, Oren Cohen, Laura Gillim, Tracy L. Hammonds, Shilpa Kapur, Harvey W. Kaufman, David Kwasny, Jean W. Liew, William A. Meyer 3rd, Shannon L. Reynolds, Cheryl B. Schleicher, Suki Subbiah, Catherine Theruviparampil, Zachary S. Wallace, Jeremy L. Warner, Suhyeon Yoon, Yonah C. Ziemba

School of Medicine Faculty Publications

Background. We developed a United States–based real-world data resource to better understand the continued impact of the coronavirus disease 2019 (COVID-19) pandemic on immunocompromised patients, who are typically underrepresented in prospective studies and clinical trials. Methods. The COVID-19 Real World Data infrastructure (CRWDi) was created by linking and harmonizing de-identified HealthVerity medical and pharmacy claims data from 1 December 2018 to 31 December 2023, with severe acute respiratory syndrome coronavirus 2 virologic and serologic laboratory data from major commercial laboratories and Northwell Health; COVID-19 vaccination data; and, for patients with cancer, 2010 to 2021 National Cancer Institute Surveillance, Epidemiology, and …

Anthropological Analysis Of The Human Skeletal Remains From Naḥalat Ahim, Jerusalem (Pp. 121–124), Joe Zias

'Atiqot

The remains of 51 individuals were recovered from a Byzantine-period tomb at Naḥalat Ahim: 47% were adults, 40% were sub-adults and 4% were between 6–12 months; no neonates were identified. There was no discernible demographic pattern regarding age, sex or place of burial within the tomb. Two benign bone tumors were found in two different individuals; both types are previously unreported in excavations in Israel.

Human Skeletal Remains From A Second -Temple-Period Tomb In Arnona, Jerusalem (Pp. 117–120), Joe Zias

'Atiqot

A minimum of 41 individuals was recovered from the tomb. The demographic profile shows that 51% of the population died before age 18, in accord with other reported figures from the period. The pathology of the interred is unique, representing the first reported incident of tuberculosis in an ancient Jewish population.

Geographic Disparities Impacting Oral Vaccine Performance: Observations And Future Directions, Rachel M Burke, Sasirekha Ramani, Julia Lynch, Laura V Cooper, Haeun Cho, Ananda S Bandyopadhyay, Carl D Kirkwood, A Duncan Steele, Gagandeep Kang

Faculty, Staff and Students Publications

Oral vaccines have several advantages compared with parenteral administration: they can be relatively cheap to produce in high quantities, easier to administer, and induce intestinal mucosal immunity that can protect against infection. These characteristics have led to successful use of oral vaccines against rotavirus, polio, and cholera. Unfortunately, oral vaccines for all three diseases have demonstrated lower performance in the highest-burden settings where they are most needed. Rotavirus vaccines are estimated to have >85% effectiveness against hospitalization in children < 12 months in countries with low child mortality, but only ~65% effectiveness in countries with high child mortality. Similarly, oral polio vaccines have lower immunogenicity in developing country settings compared with high-resource settings. Data are more limited for oral cholera vaccines, but suggest lower titers among children compared with adults, and, for some vaccines, lower efficacy in endemic settings compared with non-endemic settings. These disparities are likely multifactorial, and available evidence suggests a role for maternal factors (e.g. transplacental antibodies, breastmilk), host factors (e.g. genetic polymorphisms-with the best evidence for rotavirus-or previous infection), and environmental factors (e.g. gut microbiome, co-infections). Overall, these data highlight the rather ambiguous and often contradictory nature of evidence on factors affecting oral vaccine response, cautioning against broad extrapolation of outcomes based on one population or one vaccine type. Meaningful impact on performance of oral vaccines will likely only be possible with a suite of interventions, given the complex and multifactorial nature of the problem, and the degree to which contributing factors are intertwined.

How Did Covid-19 Impact Physiology Research Globally? – Lessons Learned And Future Recommendations, Jeet Patel, Santanu De

Mako: NSU Undergraduate Student Journal

The Coronavirus Infectious Disease 2019 (COVID-19) pandemic caused massive changes in the medical field across the globe. As the whole world moved from in-person to remote activities, students found themselves in a predicament where they were compelled towards schooling and work online from home. This would mean that all projects, research, tests, and meetings were conducted over Zoom or other online meeting platforms. Laboratories were shut down or only a limited number of people were allowed to enter to conduct the necessary research. This paper discusses how the COVID-19 pandemic affected research process and experiments within various aspects of the …

Investigating The Role Of The Nsp15 Amino-Terminal Domain In Nsp15 Function, Viral Replication, And Pathogenesis, Monika Evdokimova

Dissertations

Identifying conserved mechanisms used by viruses to delay host innate responses can reveal potential targets for antiviral therapeutics. Here, we investigated coronavirus nonstructural protein 15 (nsp15), which encodes a highly conserved endoribonuclease (EndoU). EndoU functions as an immune antagonist by limiting the accumulation of viral replication intermediates that would otherwise be sensed by the host. Despite being a promising antiviral target, it has been difficult to develop small molecule inhibitors that target the EndoU active site. We generated nsp15 mutants of SARS-CoV-2 and mouse coronavirus MHV-A59 and identified conserved residues within the amino-terminal domain that are required for EndoU activity. …

Light-Dark Shift Promotes Colon Carcinogenesis Through Accelerated Colon Aging, Deepak Sharma, Phillip A Engen, Abu Osman, Darbaz Adnan, Maliha Shaikh, Mostafa K Abdel-Reheem, Ankur Naqib, Stefan J Green, Bruce Hamaker, Christopher B Forsyth, Lin Cheng, Ali Keshavarzian, Khashayarsha Khazaie, Faraz Bishehsari

Faculty, Staff and Student Publications

Colorectal cancer (CRC) is the third most common cancer worldwide, with rising prevalence among younger adults. Several lifestyle factors, particularly disruptions in circadian rhythms by light-dark (LD) shifts, are known to increase CRC risk. Epidemiological studies previously showed LD-shifts are associated with increased risk of CRC. To explore the mechanisms and interactions between LD-shift and intestinal aging, we investigated how the combination of LD-shifts and aging impacts colon carcinogenesis development. Our data showed that LD-shifts and aging increased colon tumorigenesis. Notably, LD-shift accelerated intestinal aging by altering aging-related pathways, such as intestinal barrier damage, accompanied by dysbiotic changes in the …

Halogeton And Greasewood Poisoning In Cattle And Sheep In Utah, E. Jane Kelly, Stephen T. Lee, Ammon Brown, Daniel Cook

Poisonous Plant Research (PPR)

Halogeton (Halogeton glomeratus) is a fast-growing annual plant while greasewood (Sarcobatus vermiculatus) is a perennial shrub of the Western States. Oxalates are the toxic principle in halogeton and greasewood. Most losses occur when hungry animals are allowed to graze in heavy stands of halogeton and greasewood. Halogeton and greasewood are always dangerous and become more toxic as the growing season progresses. Here we report on two clinical cases of halogeton and greasewood poisoning in cattle and sheep. Histologically, oxalate crystals were observed in the kidneys of the cow and sheep consistent with oxalate poisoning. Oxalate was …

Hiv Associated Epigenetic Trends And Chronic Diseases: Insights Into The Hidden Burden Of Chronic Infection, Courtney G. Wallace, Zachary Capriotti, Zachary Klase

Kimmel Cancer Center Faculty Papers

Human Immunodeficiency Virus (HIV) remains a major health challenge despite dramatic advances in treatment and prevention. People living with HIV (PLWH) continue to experience high rates of non-AIDS comorbidities, including cardiovascular, renal, pulmonary, oncologic, and neurocognitive disorders. These conditions persist under viral suppression, underscoring the lasting biological impact of infection. Epigenetic dysregulation has emerged as a key driver of these outcomes. HIV integration, viral proteins, chronic inflammation, and ART exposure have all been reported to alter DNA methylation, histone modifications, transcription factor networks, and noncoding RNA regulation. These changes extend beyond infected cells, reprogramming uninfected immune and tissue compartments. Long-lived …

Exploring The Pathogen Profiles Of Ancient Feces, Drew Capone, David Holcomb, Amanda Lai, Karl Reinhard, Joe Brown

Karl Reinhard Publications

Analysis of ancient desiccated feces – termed paleofeces or coprolites – can unlock insights into the lives of ancient people. We collected desiccated feces from caves in the Rio Zape Valley in Mexico (725-920 CE). First, we extracted DNA with methods previously optimized for paleofeces. Then, we applied highly sensitive modern molecular tools (i.e., PCR pre-amplification followed by multi-parallel qPCR) to assess the presence of 30 enteric pathogens. We detected ≥1 pathogen associated gene in each of the ten samples and a mean of 3.9 pathogens per sample. The targets detected included Blastocystis spp. (n=7), atypical enteropathogenic E. coli (n=7), …

The Adoption Of Biosecurity Measures And Its Influencing Factors In Bangladeshi Layer Farms, Mohammadyar Jahangir Alam, Ismat Ara Begum, Md Al Mamun, Md Asif Iqbal, Andrew M. Mckenzie

Agricultural Economics and Agribusiness Faculty Publications and Presentations

Developing nations like Bangladesh are particularly vulnerable to poultry diseases, notably Highly Pathogenic Avian Influenza. Adopting biosecurity approaches that assist farms in disease control and prevention can decrease the industry-wide dissemination of infectious diseases. In light of these factors, this study was carried out to investigate the implementation of biosecurity practices on layer farms in Bangladesh. Using a stratified random sample method, a total of 315 layer farms from fourteen upazilas (sub-districts) in seven districts were surveyed using a semi-structured interview schedule. The study explored layer farmers' experiences with biosecurity procedures adopted on farms that included equipment, employees, and visitors. …

Tamm-Horsfall Protein Augments Neutrophil Netosis During Urinary Tract Infection, Vicki Mercado-Evans, Holly Branthoover, Claude Chew, Camille Serchejian, Alexander B Saltzman, Marlyd E Mejia, Jacob J Zulk, Ingrid Cornax, Victor Nizet, Kathryn A Patras

Faculty, Staff and Students Publications

Urinary neutrophils are a hallmark of urinary tract infection (UTI), yet the mechanisms governing their activation, function, and efficacy in controlling infection remain incompletely understood. Tamm-Horsfall glycoprotein (THP), the most abundant protein in urine, uses terminal sialic acids to bind an inhibitory receptor and dampen neutrophil inflammatory responses. We hypothesized that neutrophil modulation is an integral part of THP-mediated host protection. In a UTI model, THP-deficient mice showed elevated urinary tract bacterial burdens, increased neutrophil recruitment, and more severe tissue histopathological changes compared with WT mice. Furthermore, THP-deficient mice displayed impaired urinary NETosis during UTI. To investigate the effect of …

Characterization Of Vaccine-Enhanced Humoral Immune Responses Against Emergent Sars-Cov-2 Variants In A Convalescent Cohort, Jared Sheehan, Amber J. Trauth, Michael E. Hagensee, Alistair J. Ramsay

School of Graduate Studies Faculty Publications

Vaccination of COVID-19–convalescent individuals may generate ‘hybrid’ immunity of enhanced magnitude, durability, and cross-reactive breadth. Our primary goal was to characterize hybrid antibody (Ab) responses in a patient cohort infected with ancestral Wuhan-Hu-1 virus and vaccinated between 6 and 10 months later with the Wuhan-Hu-1–based BNT162b2 mRNA vaccine. We were particularly interested in determining the efficacy of neutralizing Ab responses against subsequently emergent SARS-CoV-2 variants. Sera collected at 3-monthly intervals over a period of 12 months were analyzed by ELISA for SARS-CoV-2 RBD–specific Ab responses, and also for neutralizing Ab activity using pseudovirus-based neutralization assays. We found that convalescent RBD-reactive …

Targeted Inhibition Of Aurora Kinase A Promotes Immune Checkpoint Inhibition Efficacy In Human Papillomavirus-Driven Cancers, Soma Ghosh, Madison P O'Hara, Pragya Sinha, Tuhina Mazumdar, Lacin Yapindi, Jagannadha K Sastry, Faye M Johnson

Faculty, Staff and Student Publications

Background: Human papillomavirus (HPV)-driven cancers include head and neck squamous cell carcinoma and cervical cancer and represent approximately 5% of all cancer cases worldwide. Standard-of-care chemotherapy, radiotherapy, and immune checkpoint inhibitors (ICIs) are associated with adverse effects and limited responses in patients with HPV-driven cancers. The integration of targeted therapies with ICIs may improve outcomes. In a previous study, we demonstrated that Aurora kinase A (AURKA, Aurora A) inhibitors lead to apoptosis of human HPV-positive cancer cells in vitro and in vivo. Here, we explored the potential of Aurora A inhibition to enhance response to ICIs in immune-competent …

Evaluation Of Green Tea Yoghurt Enriched With Lacticaseibacillus Paracasei E1 Microcapsules On Macrophage M1 Profile In High Fat-Fructose Diet Mice, Esha Ardiansyah, Nur Alfi Maghfirotus Sa’Adah, Rahmi Izati, Belinda Nabiila Al Faizah, Dawama Nur Fadlilah, Septhyanti Aprilia Kavitarna, Mochammad Fitri Atho’Illah, Siti Nur Arifah, Yoga Dwi Jatmiko, Muhaimin Rifa’I

Karbala International Journal of Modern Science

Obesity is caused by an energy imbalance that increases chronic low-grade inflammation, including macrophage cell infiltration. Adipose tissue macrophages are polarized into pro-inflammatory macrophage type 1 (M1), secrete large amounts of pro-inflammatory cytokines, and activate transcription factors. Yoghurt with probiotics is popular at all ages for its health benefits and must be protected by microencapsulation. Green tea (Camellia sinensis L.) fortification provides yoghurt nutrients while improving its functional qualities and bioactivity. This study aimed to evaluate the effect of microencapsulation of Lacticaseibacillus paracasei E1 in green tea yoghurt (GTY) on the profile of M1 macrophages in mice fed a …

Recurrent And Metastatic Head And Neck Cancer: Mechanisms Of Treatment Failure, Treatment Paradigms, And New Horizons, William T. Barham, Marshall Patrick Stagg, Rula Mualla, Michael Dileo, Sagar Kansara

School of Medicine Faculty Publications

Background: Head and neck cancer is a deadly disease with over 500,000 cases annually worldwide. Metastatic head and neck cancer accounts for a large proportion of the mortality associated with this disease. Many advances have been made in our understanding of the mechanisms of metastasis. The application of immunotherapy to locally recurrent or metastatic head and neck cancer has not only improved oncologic outcomes but has also provided valuable insights into the mechanisms of immune evasion and ultimately treatment failure. Objectives: This review paper will review our current understanding of biological mechanisms of treatment failure and metastasis. Published and ongoing …

Course Portfolio: Bios 443/843 Immunology, Nicole Sexton

UNL Faculty Course Portfolios

This course portfolio relates to the Fall 2025 delivery of BIOS 443/843: Immunology, cross listed as MBIO 443 and VMBS 843 and was prepared as part of the Faculty-led Inquiry into Reflective and Scholarly Teaching (FIRST) project. BIOS 443/843 is an upper division course that introduces Immunological concepts but also connects them to up to date literature in the field of Immunology with the intent of preparing students for careers in medicine and biomedical research. In Fall of 2025 this course was restructured to incorporate additional peer-to-peer learning opportunities and more distributed assessment styles, therefore was peer-reviewed to assess the …

Early Treatment Discontinuation In Patients With Deficient Mismatch Repair Or Microsatellite Instability High Metastatic Colorectal Cancer Receiving Immune Checkpoint Inhibitors, Julien Taieb, Margherita Ambrosini, Emily Alouani, Sara Lonardi, Frank A Sinicrope, Marie Decraecker, Alice Boileve, Emilie Hafliger, Thibault Mazard, Simon Pernot, Pauline Parent, Javier Ros, Michael J Overman, Priya Jayachandran, Vincenzo Nasca, Lisa Salvatore, Rosine Guimbaud, Chiara Cremolini, David Tougeron, Filippo Pietrantonio

Faculty, Staff and Student Publications

Background: Immune checkpoint inhibitors (ICIs) are recommended to treat patients with deficient mismatch repair/microsatellite instability high (dMMR/MSI-H) metastatic colorectal cancer (mCRC). Pivotal trials have fixed a maximum ICI duration of 2 years, without a compelling rationale. A shorter treatment duration has the potential to improve patients' quality of life and reduce both toxicity and cost without compromising efficacy. Here we examine whether early treatment discontinuation (ETD) before 13 months in patients without progressive disease (PD) can lead to similar long-term disease control compared with a longer treatment duration (LTD).

Methods: To assess whether ETD is associated with similar outcomes compared …

Fta™ Card Use For Detection Of Mycoplasma Gallisepticum In Wild Birds, Linda Purvis, Dawn E.W. Drumtra

Georgia Journal of Science

Flinders Technology Associates (FTA) cards are increasingly accepted as an accurate method for nucleic acid detection, including testing for the presence of various avian diseases. However, the accuracy of FTA cards for detection of Mycoplasma gallisepticum (MG) in wild birds has not been published. Traditional techniques for testing MG in birds include serum plate agglutanation (SPA), hemagglutination inhibition (HI), polymerase chain reaction (PCR), and enzyme linked immunosorbent assay (ELISA) all of which are time consuming and require significant financial investment into specialized equipment. We tested the efficacy of using FTA cards to detect MG in wild bird carcasses collected in …

Optimizing Biofilm Assay Reproducibility And Characterizing The 12-Hydroxystearic Acid On Escherichia Coli, Khilola Khusanova

Theses and Dissertations

Biofilm formation in Escherichia coli plays a critical role in microbial chemical communication and surface-associated persistence. This study investigates the antibiofilm activity of 12-hydroxystearic acid (12-HSA), evaluating its R and S enantiomers as well as a racemic mixture. E. coli BL21, biofilms were grown under standardized laboratory conditions and quantified using the Crystal Violet staining assay. Treatment with 12-HSA resulted in a measurable reduction in biofilm biomass, with differences observed between the individual enantiomers and the mixture. These findings suggest that 12-HSA influences early adhesion and biofilm development, supporting its potential as a small-molecule modulator of bacterial surface behavior. This …

The Effects Of Active Substances Of Clinically Important Drugs On The Survival, Development, And Some Adult Traits Of Drosophila Melanogaster Meigen (Diptera: Drosophilidae), Gökçe Üstündağ, Kemal Büyükgüzel, Ender Büyükgüzel

Turkish Journal of Zoology

Drosophila melanogaster Meigen (Diptera: Drosophilidae) was used as a model organism and fed on an artificial diet with different clinically important drugs under laboratory conditions. The effects of fluconazole, acyclovir, and piperazine on the survivorship, development, adult longevity, fecundity, and hatching rate of the insect were investigated. The results showed that the highest concentration of fluconazole (1250 mg/L) significantly decreased both the rate of larvae reaching the 3rd instar and the rate of pupation from 93.00 ± 2.95% to 11.00 ± 2.59% (χ2 = 134.696; df = 1; p = 0.0001). The highest concentration of acyclovir (200 mg/L) significantly reduced …

A Band-Aid Solution: The Need For A Better Treatment Plan For The Physician Burnout Crisis, Sarah Mangroo

Annals of Health Law and Life Sciences

No abstract provided.

Measuring Spatial And Temporal Variation In Species Composition Of Ephemeral Pond Amphibian Assemblages Using Metabarcoding, Elizabeth Strasko

Masters Theses & Specialist Projects

Amphibian populations worldwide face severe threats, heightening the need for reliable monitoring methods. In this thesis, I explored and tested environmental DNA (eDNA) metabarcoding protocols to survey amphibian assemblages in ephemeral ponds of the the Green River Preserve in central Kentucky, USA. First, I evaluated whether the commonly used mitochondrial cytochrome c oxidase subunit I (COI) gene could effectively capture species-level variation in local amphibians for eDNA metabarcoding. Extensive primer design and testing revealed significant challenges in amplifying fragmented eDNA from water samples. In a second study, I applied a metabarcoding approach using the mitochondrial 12S ribosomal RNA (12S) gene …

Characterization Of Legionella Pneumophila Effector Protein Ceg10, Emmanuella Ofoka

Masters Theses & Specialist Projects

Legionella pneumophila, the causative agent of Legionnaires' disease, utilizes a Type IV secretion system (T4SS) to transport effector proteins into host cells. This bacterium is said to have over 300 effector proteins. Among these, the Ceg10 effector protein had remained uncharacterized, despite evidence suggesting the importance of bacterial effector proteins in the host-pathogen interaction. This study aimed to perform a structural and functional characterization of Ceg10 through transcriptomic, bioinformatics, and molecular analyses. Transcriptomic profiling of HEK 293T cells expressing Ceg10 revealed significant differential gene expression, with approximately 20 upregulated genes, including leucine-rich repeat, as well as other downregulated genes. …

Atypical Stroke-Like Presentation In A Patient With Mucormycosis, Joseph Crutison, Yasser El-Daour, Dominique Pean

Gulf Coast Division GME Research Day 2025

No abstract provided.

“Science Belongs Everywhere”: Outlooks On Post-Secondary Stem Education For Correctional Centers In Virginia, Shelita R. Augustus

Division of Strategic Enrollment Management and Student Success Publications

Virginia has an incarceration rate of 679 per 100,000 residents, the highest among independent democratic nations. This scoping review examines perspectives on post-secondary STEM education to promote STEM education within Virginia's correctional facilities, assesses programs to improve rehabilitation, reduce recidivism, and address systemic inequalities by developing the scientific potential of incarcerated individuals. The review finds that while STEM education in correctional facilities faces limitations, its potential benefits are being realized nationwide. Virginia has made strides through initiatives like House Bill 2158, which aims to expand higher education access for incarcerated individuals. Examples such as the programs at J. Sargeant Reynolds …

Antimicrobial-Resistant Enterococcus In Urban Communities: Insights Into Human Infection Risks, Sumit Kumar Paudel

Wayne State University Dissertations

The complexity of community risk factors and transmission routes has been a major challenge to the containment of community-acquired antibiotic resistance. Possible factors range from environmental contamination, antibiotic misuse, and human exposure via animals, food, the environment, and human-to-human transmission. What has been missing from previous research is the integration of findings in animal and environmental sectors with bacteria implicated in community infections. To gain a better understanding of antibiotic resistance acquired in the community, a one health approach is clearly needed. Specifically, we aim to investigate the prevalence of antibiotic-resistant Enterococcus in wildlife, their gene profiles in comparison with …

Validation Of Non-Invasive Methods For The Measurement Of Gonadal And Inter-Renal Steroid Hormones In A Desert-Adapted Amphibian (Scaphiopus Couchii), Alexander T. Baugh, Callie Cho , '23, Alice Onyango-Opiyo , '23, Sophie A. Rodner , '25, Senna Mieth , '25, Daniel Oakes , '24, Liam Halstead , '25

Biology Faculty Works

For aquatic and semi-aquatic vertebrates like amphibians, it is possible to estimate excreted hormone levels using non-invasive methods such as waterborne and salivary sampling. These techniques allow monitoring of endocrine activity over varying, repeated and simultaneous integration periods while minimizing handling-related stress that can ‘contaminate’ hormone estimates, including estimates of baseline glucocorticoids. Here we have validated the extraction and quantification of three steroid hormones (corticosterone, CORT; 17-b estradiol, E₂; testosterone, TST) in Couch’s spadefoots (Scaphiopus couchii)—a desert-adapted anuran of special interest for physiology, evolution and conservation—using non-invasive waterborne and minimally invasive salivary hormone methods. We combined extraction and …