Publications by authors named "Carolyn Cray"

108 Publications

SARS-CoV-2 infection induces autoimmune antibody secretion more in lean than in obese COVID-19 patients.

medRxiv 2021 May 11. Epub 2021 May 11.

Background/objectives: Obesity decreases the secretion of SARS-CoV-2-specific IgG antibodies in the blood of COVID-19 patients. How obesity impacts the secretion of autoimmune antibodies in COVID-19 patients, however, is not understood. The serum of adult COVID-19 patients contains autoimmune antibodies generated in response to virus-induced tissue damage and cell death leading to the release of intracellular antigens not known to be immunogenic autoantigens. The objective of this study is to evaluate the presence of autoimmune antibodies in COVID-19 patients with obesity.

Subjects/methods: Thirty serum samples from individuals who tested positive for SARS-CoV-2 infection by RT-PCR were collected from inpatient and outpatient settings. Of these, 15 were lean (BMI<25), and 15 were obese (BMI ≥30). Control serum samples were from 30 uninfected individuals, age-gender- and BMI-matched, recruited before the current pandemic. Serum IgG antibodies against two autoimmune specificities, as well as against SARS-CoV-2 Spike protein, were measured by ELISA. IgG autoimmune antibodies were specific for malondialdehyde (MDA), a marker of oxidative stress and lipid peroxidation, and for adipocyte-derived protein antigens (AD), markers of virus-induced cell death in the obese AT.

Results: Our results show that SARS-CoV-2 infection induces anti-MDA and anti-AD autoimmune antibodies more in lean than in obese patients as compared to uninfected controls. Serum levels of these autoimmune antibodies, however, are always higher in obese versus lean COVID-19 patients. Moreover, because the autoimmune antibodies found in serum samples of COVID-19 patients have been correlated with serum levels of C-reactive protein (CRP), a general marker of inflammation, we also evaluated the association of anti-MDA and anti-AT antibodies with serum CRP and found a significant association between CRP and autoimmune antibodies in our cohort of lean and obese COVID-19 patients.

Conclusions: Our results highlight the importance of evaluating the quality of the antibody response in COVID-19 patients with obesity, particularly the presence of autoimmune antibodies, and identify biomarkers of self-tolerance breakdown. This is crucial to protect this vulnerable population that is at higher risk of responding poorly to infection with SARS-CoV-2 compared to lean controls.
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http://dx.doi.org/10.1101/2021.05.05.21256686DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8132267PMC
May 2021

Protein Electrophoresis and Haptoglobin Values for Captive Bongo ().

Front Vet Sci 2021 28;8:646500. Epub 2021 Apr 28.

Division of Comparative Pathology, University of Miami Miller School of Medicine, Miami, FL, United States.

Serum samples collected from 37 clinically normal bongo () and 13 abnormal bongo were tested using assays for acute-phase proteins (APPs) and by protein electrophoresis. Abnormal bongo samples ( = 27) had significantly higher levels of fibrinogen (FIB) ( < 0.001) and trending but not significantly increased haptoglobin (HP) ( = 0.07) vs. samples from normal bongo ( = 37). There were no significant differences in values for total white blood cell counts or for any of the fractions determined by protein electrophoresis. Clinically normal female bongo ( = 19) had significantly lower levels of FIB than normal males ( = 18) ( = 0.014), and this observation was also made with samples from the clinically abnormal group ( = 0.004). Many weak to moderate significant correlations were observed with increasing age, including increased globulins, FIB, and HP and decreased albumin-to-globulin (A/G) ratio and albumin. In clinical cases reviewed in this study, mild HP changes categorized this reactant as a minor APP, which contrasts with the major APP classification of HP in the related species of the cow. The preliminary data indicate that the quantitation of these APPs may offer value in assessing inflammation in this species, but additional studies are needed.
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http://dx.doi.org/10.3389/fvets.2021.646500DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8113405PMC
April 2021

Influence of obesity on serum levels of SARS-CoV-2-specific antibodies in COVID-19 patients.

PLoS One 2021 24;16(3):e0245424. Epub 2021 Mar 24.

Department of Microbiology and Immunology, University of Miami Miller School of Medicine, Miami, FL, United States of America.

SARS-CoV-2 (Severe Acute Respiratory Syndrome Corona Virus-2), cause of COVID-19 (Coronavirus Disease of 2019), represents a significant risk to people living with pre-existing conditions associated with exacerbated inflammatory responses and consequent dysfunctional immunity. In this paper, we have evaluated the influence of obesity, a condition associated with chronic systemic inflammation, on the secretion of SARS-CoV-2-specific IgG antibodies in the blood of COVID-19 patients. Our hypothesis is that obesity is associated with reduced amounts of specific IgG antibodies. Results have confirmed our hypothesis and have shown that SARS-CoV-2 IgG antibodies are negatively associated with Body Mass Index (BMI) in COVID-19 obese patients, as expected based on the known influence of obesity on humoral immunity. Antibodies in COVID-19 obese patients are also negatively associated with serum levels of pro-inflammatory and metabolic markers of inflammaging and pulmonary inflammation, such as SAA (serum amyloid A protein), CRP (C-reactive protein), and ferritin, but positively associated with NEFA (nonesterified fatty acids). These results altogether could help to identify an inflammatory signature with strong predictive value for immune dysfunction. Inflammatory markers identified may subsequently be targeted to improve humoral immunity in individuals with obesity and in individuals with other chronic inflammatory conditions.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0245424PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7990309PMC
April 2021

HEMATOLOGY, PLASMA BIOCHEMISTRY, AND PLASMA PROTEIN ELECTROPHORESIS REFERENCE INTERVALS FOR BLUE IGUANAS () FROM GRAND CAYMAN ISLAND.

J Zoo Wildl Med 2021 Jan;51(4):933-947

Wildlife Conservation Society, Zoological Health Program, Bronx, NY 10460, USA.

The blue iguana () is an endangered rock iguana species native to Grand Cayman, in the Cayman Islands. Health assessments were conducted on captive and free-roaming iguanas in 2001 and 2003-2014 and were performed in the summer wet season (June-July) of 2003-2004 and 2010-2014 and in the winter dry season (November-December) of 2001 and 2005-2009. Morphometric data were recorded from iguanas when blood samples were collected: 903 samples were collected and data from 890 samples from 775 iguanas were included. Samples were analyzed for hematology, plasma biochemistry, protein electrophoresis, mineral panels, 25-hydroxyvitamin D, and testosterone. Reference intervals were created for captive subadults, captive adults, and free-roaming adults when data were sufficient. Significant differences among these groups were described, as were differences on the basis of sex, season, and origin (captive vs free-roaming). In captive iguanas, most analytes were significantly different between subadults and adults, mature heterophils and copper were significantly higher in the dry season, zinc levels were significantly higher in the wet season, and cholesterol and triglycerides were significantly higher in adult females than adult males. Testosterone in adult males was significantly higher in the dry season. These results will aid in future health assessments and disease investigations in wild and captive populations of blue iguanas and are of comparative value for other species that are free-roaming, captive, and, especially, in similar conservation release programs.
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http://dx.doi.org/10.1638/2020-0094DOI Listing
January 2021

SERUM ACUTE-PHASE PROTEINS IN BOTTLENOSE DOLPHINS () AND CORRELATION WITH COMMONLY UTILIZED INFLAMMATORY INDICES.

J Zoo Wildl Med 2020 Nov;51(3):657-662

Chicago Zoological Society, Brookfield Zoo, Brookfield, IL 60513, USA.

Acute-phase proteins (APP) are the foundation to the innate immune response and valuable biomarkers that increase with inflammation, infection, neoplasia, stress, and trauma. Little is known about the acute-phase response in cetaceans and if these proteins can be used for health monitoring in individuals and free-ranging populations. The purpose of this study was to characterize serum concentrations of haptoglobin (Hp) and serum amyloid A (SAA), as well as electrophoretic profiles of common bottlenose dolphins () in free-ranging ( = 33) and professional care ( = 27) settings. Results were correlated to commonly utilized inflammatory indices including erythrocyte sedimentation rate (ESR), fibrinogen, total white blood cell count (WBC), and absolute neutrophil count. SAA levels, measured with a dolphin-specific enzyme linked immunosorbent assay (ELISA), were significantly higher ( = 0.05) in free-ranging dolphins (mean = 4.26; SE = 1.12) when compared with those under professional care (mean = 1.82; SE = 0.45). For dolphins under professional care, a statistically significant correlation was identified between ESR and Hp ( < 0.001; = 0.69), ESR and SAA ( < 0.001; = 0.67), fibrinogen and Hp ( = 0.001; = 0.58), and fibrinogen and SAA ( = 0.002; = 0.56). In addition, there was a significant correlation between WBC and SAA ( = 0.01; = 0.38) and absolute neutrophil count and SAA ( = 0.04; = 0.32). There were no significant correlations between study variables observed in free-ranging dolphins. The variable correlation of APPs with commonly utilized inflammatory indices demonstrates that these proteins are independent measures of inflammation with unique sensitivity, specificity, and timeline of expression. The results of this study contribute to improved health monitoring of dolphins and have the potential to assist in identification of compromised health.
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http://dx.doi.org/10.1638/2019-0130DOI Listing
November 2020

PLASMA BIOCHEMISTRY AND PROTEIN ELECTROPHORESIS REFERENCE INTERVALS OF THE COMMON LOON ().

J Zoo Wildl Med 2020 Nov;51(3):561-570

Tufts Cummings School of Veterinary Medicine, North Grafton, MA 01536, USA.

There are no published plasma biochemistry reference intervals for any species within the order Gaviiformes, which includes the common loon (). Because of their unique classification and lack of close taxonomic relatives, species-specific values for clinical data in loons are needed. This study determined reference intervals for plasma biochemical values in adult common loons, and reference intervals for protein electrophoresis values in both adult and juvenile common loons. Healthy, wild adult ( = 148, age >3 yr) and juvenile ( = 31, age 4-12 wk) common loons were sampled on freshwater summer breeding territories at study sites across North America. Plasma biochemical analytes included glucose (Glu), total calcium, phosphorus, sodium, potassium, chloride, blood urea nitrogen (BUN), uric acid, cholesterol, triglycerides, creatine kinase, γ-glutamyl transferase, alkaline phosphatase (ALP), lactate dehydrogenase, aspartate aminotransferase, amylase, and bile acids. Protein electrophoresis data included albumin to globulin ratio (A: G), prealbumin, albumin, α1-globulin, α2-globulin, β-globulin, and γ-globulin. Adult females had significantly higher Glu, ALP, and BUN than adult males. Juvenile loons had higher β-globulins than adults, whereas adults had higher α1-globulins. Establishment of complete reference intervals will improve clinical assessment of captive loons, and allow researchers to better understand the health of wild loons in response to the multiple environmental stressors faced by these species.
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http://dx.doi.org/10.1638/2019-0168DOI Listing
November 2020

Effects of obesity on serum levels of SARS-CoV-2-specific antibodies in COVID-19 patients.

medRxiv 2020 Dec 20. Epub 2020 Dec 20.

SARS-CoV-2 (Severe Acute Respiratory Syndrome Corona Virus-2), cause of COVID-19 (Coronavirus Disease of 2019), represents a significant risk to people living with pre-existing conditions associated with exacerbated inflammatory responses and consequent dysfunctional immunity. In this paper, we have evaluated the effects of obesity, a condition associated with chronic systemic inflammation, on the secretion of SARS-CoV-2-specific IgG antibodies in the blood of COVID-19 patients. Results have shown that SARS-CoV-2 IgG antibodies are negatively associated with Body Mass Index (BMI) in COVID-19 obese patients, as expected based on the known effects of obesity on humoral immunity. Antibodies in COVID-19 obese patients are also negatively associated with serum levels of pro-inflammatory and metabolic markers of inflammaging and pulmonary inflammation, such as SAA (serum amyloid A protein), CRP (C-reactive protein) and ferritin, but positively associated with NEFA (nonesterified fatty acids). These results altogether could help to identify an inflammatory signature with strong predictive value for immune dysfunction that could be targeted to improve humoral immunity in individuals with obesity as well as with other chronic inflammatory conditions.
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http://dx.doi.org/10.1101/2020.12.18.20248483DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7783955PMC
December 2020

Capillary Electrophoresis Assessment of Plasma Protein Changes in an African Penguin () With Aspergillosis.

ACS Omega 2020 Dec 15;5(51):33280-33289. Epub 2020 Dec 15.

Miami Integrative Metabolomics Research Center, University of Miami, Miami, Florida 33136, United States.

A decrease of avian biodiversity in the African continent has been the result of anthropogenic pressure in the region. This has resulted in the African penguin () being placed on the endangered species list and requires conservation efforts to maintain its free-ranging population and placement under managed care. In the latter environment, infection by can be common. The diagnosis and treatment of this fungal disease in birds has presented with many difficulties, largely due to the diversity and limited knowledge that exists about this species. In this study, we implement a high-resolution capillary electrophoresis system for the fractionation of African penguin plasma, followed by mass spectrometry analysis for the identification of proteins associated with aspergillosis. Several protein differences were revealed, including changes in acute phase proteins and lipid metabolism. In addition, our results demonstrated that fibrinogen β chain is a protein largely present during the inflammatory process in an African penguin infected with . These findings present a new avenue for the measurement of plasma proteins as a potential method for identifying important biomarkers to aid in monitoring African penguin health.
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http://dx.doi.org/10.1021/acsomega.0c04983DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7774288PMC
December 2020

The importance of manual white blood cell differential counts and platelet estimates in elephant hematology: blood film review is essential.

Vet Q 2021 Dec;41(1):30-35

Department of Comparative, Diagnostic, and Population Medicine, College of Veterinary Medicine, University of Florida, Gainesville, FL, USA.

Unique features of elephant hematology are known challenges in analytical methodology like two types of monocytes typical for members of the Order Afrotheria and platelet counts of the comparatively small elephant platelet. To investigate WBC differential and platelet data generated by an impedance-based hematology analyzer without availability of validated species-specific software for recognition of elephant WBCs and platelets, compared to manual blood film review. Blood samples preserved in ethylenediaminetetraacetic acid (EDTA) of 50 elephants (n = 35 and n = 15 ) were used. A Mann-Whitney test for independent samples was used to compare parameters between methods and agreement was tested using Bland-Altman bias plots. All hematological variables, including absolute numbers of heterophils, lymphocytes, monocytes, eosinophils, basophils, and platelets, were significantly different ( < 0.0001) between both methods of analysis, and there was no agreement using Bland-Altman bias plots. Manual review consistently produced higher heterophil and monocyte counts as well as platelet estimates, while the automated analyzer produced higher lymphocyte, eosinophil, and basophil counts. The hematology analyzer did not properly differentiate elephant lymphocytes and monocytes, and did not accurately count elephant platelets. These findings emphasize the importance of manual blood film review as part of elephant complete blood counts in both clinical and research settings and as a basis for the development of hematological reference intervals.
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http://dx.doi.org/10.1080/01652176.2020.1867329DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7817171PMC
December 2021

Blood analytes of immature Kemp's ridley sea turtles () from Georgia, USA: reference intervals and body size correlations.

Conserv Physiol 2020 1;8(1):coaa091. Epub 2020 Dec 1.

Aquatic, Amphibian, and Reptile Pathology Program, Department of Comparative, Diagnostic, and Population Medicine, College of Veterinary Medicine, University of Florida, Gainesville, Florida, 32608, USA.

Health assessments of wildlife species are becoming increasingly important in an ever-changing environment. Kemp's ridley sea turtles (; hereafter, Kemp's ridleys) are critically endangered and incur several on-going threats to their population recovery; therefore, it is imperative to advance the understanding of baseline blood analyte data as a diagnostic and monitoring tool. For in-water, trawl-captured, immature Kemp's ridleys (minimum  = 31) from Georgia, USA, the objectives of this study were to (1) establish reference intervals (RIs) for packed cell volume (PCV) and 27 plasma biochemistry analytes and (2) determine length-specific relationships in blood analytes. We observed significant positive correlations between minimum straight carapace length and PCV, amylase, calcium:phosphorus ratio, cholesterol, magnesium, triglycerides, total solids, total protein and all protein fractions (e.g. alpha-, beta- and gamma-globulins); aspartate aminotransferase and chloride showed significant negative relationships. These results suggest that certain blood analytes in Kemp's ridleys change as these animals grow, presumptively due to somatic growth and dietary shifts. The information presented herein, in due consideration of capture technique that may have impacted glucose and potassium concentrations, represents the first report of blood analyte RIs for Kemp's ridley sea turtles established by guidelines of the American Society for Veterinary Clinical Pathology and will have direct applications for stranded individuals in rehabilitative care and for future investigations into the health status of wild individuals from this population.
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http://dx.doi.org/10.1093/conphys/coaa091DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7720087PMC
December 2020

The Iconic Atlantic Goliath Grouper (): A Comprehensive Assessment of Health Indices in the Southeastern United States Population.

Front Vet Sci 2020 25;7:635. Epub 2020 Sep 25.

Division of Comparative Pathology, Department of Pathology & Laboratory Medicine, University of Miami Miller School of Medicine, Miami, FL, United States.

The Atlantic Goliath Grouper () population has rebounded from near extinction to an international status as vulnerable due in part to regional species recovery efforts. The southeastern US population has been recovering with the main spawning locations off the coasts of Florida. Despite their economic importance to the catch-and-release fishery and the dive industry, and their ecological importance as ecosystem engineers resulting in positive impacts on reefs and species richness, baseline health assessment information is very limited in this species to date. The objectives of this study were to: (1) establish reference intervals for hematological and plasma biochemical analytes, and report immune function, oxidative stress, and vitellogenin in mature males and females; (2) evaluate total length, age, and sex in relation to blood analytes in juvenile and mature fish; (3) assess analytes across sampled months in mature male and female fish; and (4) describe the typical light microscopy findings in liver and gill biopsies, including quantitative assessment of pigmented macrophage aggregates. Health indices are reported as reference intervals when applicable, or otherwise descriptively. Blood analyte correlations with length and age, sex differences, and comparisons across months provided relevant physiological considerations, including differences in protein/energy metabolism, tissue growth, sexual maturation, active reproduction, and antigenic stimulation. Liver histology identified changes associated with life stage, active reproduction, or of subclinically to clinically insignificant infectious and/or inflammatory processes. Hepatocellular vacuolation and pigmented macrophage aggregates were prominent. Pigmented macrophage aggregates correlated with total length, presumably from continuous antigenic stimulation and/or metabolic changes as fish grow. Gill histological findings were subtle. The data presented herein provide an essential baseline assessment of a suite of health variables in an iconic marine teleost species, serves as a springboard for future studies relevant to conservation physiology, and allows for population-level applications for conservation management and policy.
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http://dx.doi.org/10.3389/fvets.2020.00635DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7546827PMC
September 2020

A How-to Guide to Building a Robust SARS-CoV-2 Testing Program at a University-Based Health System.

Acad Pathol 2020 Jan-Dec;7:2374289520958200. Epub 2020 Sep 30.

Department of Medicine, University of Miami Miller School of Medicine, Miami, FL, USA.

When South Florida became a hot spot for COVID-19 disease in March 2020, we faced an urgent need to develop test capability to detect SARS-CoV-2 infection. We assembled a transdisciplinary team of knowledgeable and dedicated physicians, scientists, technologists, and administrators who rapidly built a multiplatform, polymerase chain reaction- and serology-based detection program, established drive-through facilities, and drafted and implemented guidelines that enabled efficient testing of our patients and employees. This process was extremely complex, due to the limited availability of needed reagents, but outreach to our research scientists and multiple diagnostic laboratory companies, and government officials enabled us to implement both Food and Drug Administration authorized and laboratory-developed testing-based testing protocols. We analyzed our workforce needs and created teams of appropriately skilled and certified workers to safely process patient samples and conduct SARS-CoV-2 testing and contact tracing. We initiated smart test ordering, interfaced all testing platforms with our electronic medical record, and went from zero testing capacity to testing hundreds of health care workers and patients daily, within 3 weeks. We believe our experience can inform the efforts of others when faced with a crisis situation.
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http://dx.doi.org/10.1177/2374289520958200DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7545514PMC
September 2020

Predicting Disease Severity and Outcome in COVID-19 Patients: A Review of Multiple Biomarkers.

Arch Pathol Lab Med 2020 12;144(12):1465-1474

The Department of Pathology & Laboratory Medicine, University of Miami Miller School of Medicine, Miami, Florida (Cray).

Context.—: An abundance of clinical reports focused on specific laboratory parameters have been reported on coronavirus disease 19 (COVID-19), but a systematic analysis synthesizing these findings has not been performed.

Objective.—: To review and summarize the current available literature on the predictive role of various biomarkers in COVID-19 patients.

Data Sources.—: A literature search was performed using databases including PubMed, medRxiv, and bioRxiv. A total of 72 papers were reviewed, including 54 peer-reviewed papers and 18 non-peer-reviewed preprints.

Conclusions.—: Although the markers are considered nonspecific, acute-phase reactants, including C-reactive protein (CRP), ferritin, serum amyloid A (SAA), and procalcitonin, were reported as sensitive markers of acute COVID-19 disease. Significantly elevated white blood cell count; marked lymphopenia; decreased CD3, CD4, or CD8 T-lymphocyte counts; high neutrophil count; thrombocytopenia; and markedly elevated inflammatory biomarkers were associated with severe disease and the risk of developing sepsis with rapid progression. Trends observed by serial laboratory measurements during hospitalization, including progressive decrease of lymphocyte count, thrombocytopenia, elevated CRP, procalcitonin, increased liver enzymes, decreased renal function, and coagulation derangements, were more common in critically ill patient groups and associated with a high incidence of clinical complications. Elevated interleukin 6 level and markedly increased SAA were most often reported in severely and critically ill patients. Indicators of systemic inflammation, such as neutrophil to lymphocyte ratio, systemic immune-inflammation index, or COVID-19 Severity Score, may be used to predict disease severity, outcome, and mortality. Interpretation of the data reported in the studies reviewed here is limited because of the study design (mostly retrospective), limited sample size, and a lack of defined clinical criteria.
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http://dx.doi.org/10.5858/arpa.2020-0471-SADOI Listing
December 2020

Review of Current Advances in Serologic Testing for COVID-19.

Am J Clin Pathol 2020 08;154(3):293-304

Department of Pathology and Laboratory Medicine, University of Miami Miller School of Medicine and Jackson Health System, Miami, FL.

Objectives: To examine and summarize the current literature on serologic methods for the detection of antibodies to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).

Methods: A literature review was performed using searches in databases including PubMed, medRxiv, and bioRxiv. Thirty-two peer-reviewed papers and 23 preprints were examined.

Results: The studies included lateral flow immunoassay, enzyme-linked immunosorbent assay, chemiluminescence immunoassay, and neutralizing antibody assays. The use of all major SARS-CoV-2 antigens was demonstrated to have diagnostic value. Assays measuring total antibody reactivity had the highest sensitivity. In addition, all the methods provided opportunities to characterize the humoral immune response by isotype. The combined use of IgM and IgG detection resulted in a higher sensitivity than that observed when detecting either isotype alone. Although IgA was rarely studied, it was also demonstrated to be a sensitive marker of infection, and levels correlated with disease severity and neutralizing activity.

Conclusions: The use of serologic testing, in conjunction with reverse transcription polymerase chain reaction testing, was demonstrated to significantly increase the sensitivity of detection of patients infected with SARS-CoV-2. There was conflicting evidence regarding whether antibody titers correlated with clinical severity. However, preliminary investigations indicated some immunoassays may be a surrogate for the prediction of neutralizing antibody titers and the selection of recovered patients for convalescent serum donation.
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http://dx.doi.org/10.1093/ajcp/aqaa112DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7337672PMC
August 2020

ESTABLISHMENT OF ACUTE-PHASE PROTEIN AND SERUM PROTEIN ELECTROPHORESIS PRELIMINARY REFERENCE VALUES FOR PRONGHORN ().

J Zoo Wildl Med 2020 Jun;51(2):321-325

Division of Comparative Pathology, Department of Pathology & Laboratory Medicine, University of Miami School of Medicine, Miami, FL 33136, USA.

Pronghorn () are native to western North America and are found in 24 Association of Zoos and Aquariums (AZA)-accredited institutions. Acute-phase proteins (APP) are a broad class of proteins that are stimulated in response to inflammation and have been shown to be a sensitive measure of inflammation in equids and ruminants. In this study, blood samples from clinically normal free-ranging and captive populations of pronghorn were analyzed using assays for protein electrophoresis (EPH) and APP, including serum amyloid A (SAA) and haptoglobin (HP), to develop preliminary ranges to gauge potential differences between these populations. Additional samples were taken from clinically abnormal captive pronghorn with facial abscesses. By EPH measurements, albumin: globulin ratio mean and SE were significantly different ( <0.05) with 1.02 (0.08) for captive populations and 1.91 (0.05) for free-ranging populations. Total protein mean and SE were significantly different ( <0.05) for captive and free-ranging populations, respectively 5.6 (0.3) g/dl and 6.9 (0.1) g/dl. Mean and SD of SAA for captive pronghorn were 1.4 (3.2) mg/L, and were significantly different from the free-ranging population, which was below the limits of detection for ( <0.05). There was no difference in HP levels between these groups. In a case study of a pronghorn with facial abscesses, elevated levels of HP, but not SAA, suggested that HP maybe useful in certain disease states. Future studies should explore the use of these biomarkers as tools to monitor general health, prognosis, and subclinical disease.
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http://dx.doi.org/10.1638/2018-0226DOI Listing
June 2020

COMPARISON OF AGAROSE GEL AND CAPILLARY ZONE ELECTROPHORESIS METHODS USING PLASMA FROM GREEN TURTLES ().

J Zoo Wildl Med 2020 Mar;51(1):123-130

Division of Comparative Pathology, Department of Pathology and Laboratory Medicine, University of Miami Miller School of Medicine, PO Box 016960 (R-46), Miami, Florida 33101, USA,

Agarose gel electrophoresis (AGE) has been widely implemented throughout veterinary medicine and for analysis of plasma proteins of avian and reptile species. Capillary zone electrophoresis (CZE) is becoming a standard method in human clinical pathology laboratories but has not widely been used for the analysis of animal samples. The objective of the present study was to compare protein fractions derived from AGE and CZE methods using plasma from the green turtle (). Plasma samples were analyzed by AGE and CZE per manufacturer guidelines. The methods were assessed by CV analysis, Spearman's correlation, Passing-Bablok regression, and Bland Altman plots. CZE consistently resolved more fractions than AGE with three fractions observed in the prealbumin migrating region versus one for AGE and two fractions in the γ globulin region versus one for AGE. Compared with AGE, CZE showed a lower CV in intra-assay tests (1.0-4.9% vs 2.0-28.3%) and a lower or overlapping CV in interassay tests (1.0-10.6 vs 2.3-22.0). The prealbumin, α2 globulin, and β globulin fractions correlated the least between the methods (for all three fractions: ≤ 0.28, > 0.21). Moderate, significant correlations between AGE and CZE methods were observed for albumin ( = 0.78, < 0.0001) and γ globulins ( = 0.78, < 0.0001). CZE has a higher precision and ease of use over AGE and offers the opportunity to resolve additional protein fractions. This will necessitate the development of new conventions in placement of fraction delimits, definition of species-specific reference intervals, and evaluation of clinical utility in abnormal turtles.
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http://dx.doi.org/10.1638/2019-0120DOI Listing
March 2020

Serum Protein Analysis of Nurse Sharks.

J Aquat Anim Health 2020 06 11;32(2):77-82. Epub 2020 Mar 11.

Department of Marine Ecosystems and Society, Rosenstiel School of Atmospheric Science, University of Miami, 4600 Rickenbacker Causeway, Miami, Florida, 33149, USA.

Serum protein electrophoresis (EPH) is used to assess relative concentrations of blood proteins in clinical and biological studies. Serum EPH fractions have been determined for elasmobranchs using mammalian albumin, alpha 1-, alpha 2-, beta-, and gamma-globulin fractions, and have been deemed fractions 1 through 5, respectively. However, serum EPH fraction concentration reference intervals (RIs) have not been widely established for different elasmobranch species. In this study, RIs for fractions 1 through 5 were determined from 45 wild-caught Nurse Sharks Ginglymostoma cirratum (27 females and 23 males) in South Florida. Serum samples were isolated from whole blood following caudal venipuncture. Body condition was also measured in the field to assess the relative health of the individuals sampled. There was no relationship between body condition and serum EPH fraction concentrations. In addition, there was no difference in body condition or serum EPH fraction concentrations between females and males. Total solids and total protein values were significantly different (P < 0.001). Nurse Shark serum EPH fraction 1 was found within the mammalian albumin migrating band distance and was negligible. Fraction 2 showed no peak in the mammalian alpha 1-globulin range. A thin, medium peak in the mammalian alpha 2-globulin range represented fraction 3. In the mammalian beta-globulin range, fraction 4 consisted of the majority of protein observed. It was represented by a smooth, broad peak. A short, medium broad peak in the mammalian gamma-globulin range represented fraction 5. The Nurse Shark serum EPH fraction RIs provided in this study may be utilized to clinically evaluate the health of Nurse Sharks in captivity and in the wild, and to compare the health of their populations around the world experiencing various anthropogenic stressors and other environmental impacts.
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http://dx.doi.org/10.1002/aah.10100DOI Listing
June 2020

Assessment of the Acute Phase Response in Healthy and Injured Southern White Rhinoceros (C).

Front Vet Sci 2019 9;6:475. Epub 2020 Jan 9.

Division of Molecular Biology and Human Genetics, Department of Science and Technology-National Research Foundation Centre of Excellence for Biomedical TB Research, Faculty of Medicine and Health Sciences, Medical Research Council Centre for Tuberculosis Research, Stellenbosch University, Stellenbosch, South Africa.

Acute phase reactants (APRs) have not been investigated in white rhinoceros (). This study aimed to identify clinically useful APRs in this species. Reference intervals (RIs) were generated for albumin, fibrinogen, haptoglobin, iron and serum amyloid A (SAA) from 48 free-ranging animals, except for SAA ( = 23). APR concentrations between healthy animals and those with tissue injury (inflammation) ( = 30) were compared. Diagnostic performance was evaluated using receiver-operator characteristic (ROC) curve and logistic regression analyses. RIs were: albumin 18-31 g/L, fibrinogen 1.7-2.9 g/L, haptoglobin 1.0-4.3 g/L, iron 9.7-35.0 μmol/L, SAA <20 mg/L. Iron and albumin were lower and fibrinogen, haptoglobin and SAA higher in injured vs. healthy animals. Iron showed the best diagnostic accuracy followed by fibrinogen, albumin, haptoglobin and SAA. Iron ≤ 15.1 μmol/L and haptoglobin >4.7 g/L were significant predictors of inflammatory status and together correctly predicted the clinical status of 91% of cases. SAA > 20 mg/L had a specificity of 100%. In conclusion, albumin and iron are negative and fibrinogen, haptoglobin and SAA positive APRs in the white rhinoceros. The combination of iron and haptoglobin had an excellent diagnostic accuracy for detecting inflammation.
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http://dx.doi.org/10.3389/fvets.2019.00475DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6962144PMC
January 2020

Bias between two methods of albumin measurement in the white rhinoceros, Ceratotherium simum.

Vet Clin Pathol 2020 Mar 10;49(1):91-94. Epub 2020 Jan 10.

Department of Companion Animal Clinical Studies and Centre for Veterinary Wildlife Studies, Faculty of Veterinary Science, University of Pretoria, Pretoria, South Africa.

Background: The bromocresol green (BCG) method has been reported to overestimate serum albumin concentration in several species due to non-specific binding to globulins. As the white rhinoceros has high concentrations of serum globulins, significant differences in albumin measured by the BCG method, and the field method of agarose gel serum protein electrophoresis (SPE) are expected.

Objectives: We aimed to compare the BCG and SPE methods for albumin determination in the serum of white rhinoceroses.

Methods: SPE and BCG albumin were measured in 82 white rhinoceros serum samples. Results were compared using Bland-Altman difference plots and Passing-Bablok regression analysis.

Results: BCG albumin showed a significant mean constant positive bias of 7 g/L, or 36%, which was more than the total allowable error of 15% and was clinically significant. Methods were not comparable within the inherent imprecision of each method.

Conclusions: The BCG method overestimated albumin concentrations in this species compared with agarose gel SPE, and method-specific reference intervals should be used.
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http://dx.doi.org/10.1111/vcp.12820DOI Listing
March 2020

Spotlight on avian pathology: aspergillosis.

Avian Pathol 2020 Apr 10;49(2):115-118. Epub 2019 Dec 10.

Poultry Diagnostic and Research Center, Department of Population Health, University of Georgia, Athens, GA, USA.

Aspergillosis is a disease of domestic and free-living birds caused by infection with a fraction of fungi within the genus . Species can be identified by colony morphology and microscopic characterization of conidia and conidiophores or by PCR, and isolates can be typed by microsatellite typing. Serodiagnostic options for testing are limited to antibody and antigen detection methods. The sensitivity of these tests can be enhanced through the use of protein electrophoresis. In many countries, no systemic antifungal drugs are registered for use in food-producing birds and resistance to antifungal drugs has been reported in strains derived from birds. The most important prevention strategy against aspergillosis is keeping the infection pressure low by adequate ventilation as well as cleaning and disinfection. Open questions about avian aspergillosis that research needs to address are related to epidemiology and serodiagnosis, as well as therapy and prevention.
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http://dx.doi.org/10.1080/03079457.2019.1696946DOI Listing
April 2020

Plasma chemistry in nesting leatherback sea turtles (Dermochelys coriacea) from Florida: Understanding the importance of sample hemolysis effects on blood analytes.

PLoS One 2019 10;14(9):e0222426. Epub 2019 Sep 10.

Loggerhead Marinelife Center, Juno Beach, Florida, United States of America.

Plasma chemistry is widely used in diagnostic and research settings in sea turtles. However, plasma discolorations such as hemolysis are often not considered in data interpretation. The objectives of this study were to (1) evaluate the effects of moderate hemolysis on plasma electrolytes, minerals, and proteins using dry chemistry analysis (DCA) and protein electrophoresis from nesting leatherback sea turtles (Dermochelys coriacea) from Florida and to (2) establish blood analyte reference intervals. Twenty-six plasma samples with absence of hemolysis were selected and sub-divided into one non-hemolytic aliquot and an aliquot that was experimentally manipulated to mimic moderate hemolysis. Plasma samples were analyzed for hemoglobin using a handheld photometer; sodium, potassium, chloride, magnesium, calcium, phosphorus, and total protein using DCA; and protein electrophoresis. Packed cell volume and hemoglobin were measured in corresponding whole blood samples. Reference intervals were established. All analytes except calcium and pre-albumin were significantly higher and the calcium:phosphorus and albumin:globulin ratios were significantly lower in hemolytic plasma compared to non-hemolytic plasma. Alpha2-globulins and potassium were the analytes most impacted by hemolysis, averaging 3.3- and 2.0-fold higher in hemolyzed samples, respectively, indicating that (1) hemoglobin migrates into the alpha2-globulin region in this species and (2) notable intracellular potassium is released into plasma with hemolysis. Attempted conversion factors for compensation of hemolysis were considered inaccurate for 4 of 16 analytes due to non-significant regression lines. We also report that PCV provides an estimate of hemoglobin (g/L) using the formula: (2.59 × PCV) + 24.59. Given the spurious effects of hemolysis, the degree of this artifact should be reported with biochemistry data, and samples with moderate to severe hemolysis should be excluded from datasets when interpreting electrolyte, mineral, and protein results. This will ensure accurate data interpretation for individual turtles in rehabilitation or research settings and population-level data relevant to conservation-focused projects.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0222426PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6736308PMC
March 2020

REFERENCE INTERVALS FOR ACUTE PHASE PROTEINS FOR KOALAS () AT THE SAN DIEGO ZOO.

J Zoo Wildl Med 2019 Sep;50(3):735-738

University of Miami Miller School of Medicine, Department of Pathology and Laboratory Medicine, Miami, Florida 33136, USA.

The synthesis and circulating concentrations of acute phase proteins (APPs) are regulated in response to inflammation, infection, trauma, and neoplasia in many domestic and nondomestic species. The APP response is species specific; thus, assays must be validated, and reference intervals must be determined for each species. Koalas () are a vulnerable species, threatened by infectious and inflammatory diseases both under human care and in the wild. The ability to diagnose, treat, and provide prognosis for common koala health problems is challenged by the paucity of sensitive diagnostic tests. Assays for C-reactive protein, serum amyloid A, and haptoglobin were validated for use in koalas. Reference intervals were established using the robust method recommended by the American Society for Veterinary Clinical Pathology based on serum samples from 26 healthy koalas at the San Diego Zoo. The reference intervals are as follows: C-reactive protein, 3.2-24.1 mg/L; serum amyloid A, 0.10-0.45 mg/L; haptoglobin, 0.10-0.64 mg/ml.
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http://dx.doi.org/10.1638/2018-0227DOI Listing
September 2019

Labeled quantitative mass spectrometry to study the host response during aspergillosis in the common bottlenose dolphin (Tursiops truncatus).

Vet Microbiol 2019 May 30;232:42-49. Epub 2019 Mar 30.

University of Miami, Division of Comparative Pathology, Department of Pathology & Laboratory Medicine, Miller School of Medicine, Miami, FL, 33136, USA. Electronic address:

Aspergillosis is a fungal infection caused by Aspergillus molds that can affect both humans and animals. Despite advances in diagnostics and therapy, medical management of this disease remains difficult. Expansion of the basic knowledge regarding its pathophysiology in animals is critical to aid in the identification of new biomarkers of infection for diagnosis and therapeutic targets. For such a purpose, proteomics can be used by addressing protein changes during various disease processes. In the present study, a mass spectrometry analysis based on isobaric tagging for relative and absolute quantitation (iTRAQ) was applied for direct identification and relative quantitation of proteins in blood collected from 32 Aspergillus-diseased common bottlenose dolphins (Tursiops truncatus, 32 samples) in comparison with blood from 55 other dolphins (55 samples from 41 clinically-normal controls and from 14 cetaceans with miscellaneous non-Aspergillus inflammation diseases) and ten convalescent dolphins (28 samples). Sixty-six and 40 proteins were found to be ≥2.0-fold over- and underrepresented versus miscellaneous non-Aspergillus inflammatory dolphins, respectively, and most were confirmed vs. clinically-normal controls and convalescents. Many proteins which play a role in the adaptive immune response were identified, including MHC proteins and others involved in catalytic activity like the NADPH-ubiquinone oxido-reductases. Overall, iTRAQ appears to be a convenient proteomic tool greatly suited for exploratory ex vivo studies focusing on pathophysiology. This technique should be considered as a preliminary step before validation of new diagnostic markers.
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http://dx.doi.org/10.1016/j.vetmic.2019.03.030DOI Listing
May 2019

Serum amyloid A and plasma protein electrophoresis fractions in farmed white-tailed deer.

J Vet Diagn Invest 2019 May 11;31(3):458-462. Epub 2019 Mar 11.

Division of Comparative Pathology, Department of Pathology & Laboratory Medicine, University of Miami School of Medicine, Miami, FL (Cray).

Tools to measure the acute-phase response have been utilized widely in veterinary medicine. Evaluation by plasma protein electrophoresis (PPEP) has become an increasingly common assay in veterinary clinical pathology. Commercial reagents for serum amyloid A (SAA) have been validated for use in a variety of wildlife species. We analyzed samples from 29 healthy fawns and 60 healthy adult farmed white-tailed deer (WTD; Odocoileus virginianus) using an automated assay for SAA and a semi-automated method for PPEP. The robust statistical method for reference interval generation was used. SAA levels in fawns (0.1-26 mg/L) were found to be significantly higher than those in adults (0.1-5 mg/L, p < 0.01). The mean total protein was significantly lower in fawns (48 ± 10 g/L, p < 0.01) than in adults (73 ±5 g/L). The albumin-to-globulin ratio was also lower in fawns (0.56 ± 0.14) than in adults (1.25 ± 0.19, p < 0.01). Changes in SAA levels were observed in a variety of clinically abnormal animals. The combined use of the automated and semi-automated assays in our study may provide an additional valuable assessment tool in the care of captive WTD populations, for research studies, and for monitoring free-ranging animals.
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http://dx.doi.org/10.1177/1040638719836150DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6838698PMC
May 2019

DIAGNOSTIC PERFORMANCE OF INFLAMMATORY MARKERS IN GOPHER TORTOISES ( GOPHERUS POLYPHEMUS).

J Zoo Wildl Med 2018 Sep;49(3):765-769

The lack of sensitive and specific markers of inflammation poses a diagnostic challenge in sick or injured reptile patients. The objective of this study was to investigate the diagnostic performance of blood analytes associated with inflammation in healthy ( n = 24) and sick ( n = 38) gopher tortoises ( Gopherus polyphemus). Receiver operating characteristic analysis identified the following as the best-performing diagnostic tests: erythrocyte sedimentation rate (area under the curve [AUC] = 0.812; 95% confidence interval [CI] = 0.693, 0.900), absolute mature heterophils (AUC = 0.771; 95% CI = 0.646, 0.869), total leukocytes (AUC = 0.767; 95% CI = 0.642, 0.866), lactate (AUC = 0.766; 95% CI = 0.641, 0.864), and absolute immature heterophils (AUC = 0.755; 95% CI = 0.628, 0.856). These results support the clinical application of additional tools for the diagnosis and monitoring of inflammatory disease in gopher tortoises. Clinicians may consider adding erythrocyte sedimentation rate and lactate to the minimum database for this species.
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http://dx.doi.org/10.1638/2017-0211.1DOI Listing
September 2018

APPLICATION OF 3-HYDROXYBUTYRATE MEASUREMENT AND PLASMA PROTEIN ELECTROPHORESIS IN THE DIAGNOSIS OF ASPERGILLOSIS IN AFRICAN PENGUINS ( SPHENISCUS DEMERSUS).

J Zoo Wildl Med 2018 Sep;49(3):696-703

New alternative laboratory means are needed to improve the options for antemortem diagnosis of avian aspergillosis. In this study, 3-hydroxybutyrate was measured in plasma samples collected from a cohort of African penguins ( Spheniscus demersus) maintained under human care. Results were interpreted in combination with those of protein electrophoresis and compared with anti- Aspergillus antibody and galactomannan antigen detection. Overall, 3-hydroxybutyrate levels were found significantly increased in Aspergillus-diseased cases versus the control penguin group ( P = 0.002). Mean absolute concentration of β-globulins was increased >20% in samples from infected birds, and α2-globublins were also found to be significantly increased versus clinically normal controls ( P < 0.001 and P = 0.001 respectively). Of note, the α2-globulins were also significantly increased versus penguins with inflammatory (non-aspergillosis) diseases ( P = 0.001). The specificity of 3-hydroxybutyrate, β-globulins, and α2-globulins for aspergillosis was 78.6%, 79.6%, and 92.2%, respectively. Using these measures in tandem resulted in high specificity (>90%) and negative predictive value (≥80%). In contrast, anti- Aspergillus antibody and galactomannan antigen did not distinguish between infected cases and controls ( P > 0.05). This study demonstrates that basic testing in tandem with the new biomarker 3-hydroxybutyrate may provide reliable evidence for the diagnosis of aspergillosis in penguins.
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http://dx.doi.org/10.1638/2017-0172.1DOI Listing
September 2018

REFERENCE INTERVALS FOR ERYTHROCYTE SEDIMENTATION RATE, LACTATE, FIBRINOGEN, HEMATOLOGY, AND PLASMA PROTEIN ELECTROPHORESIS IN CLINICALLY HEALTHY CAPTIVE GOPHER TORTOISES ( GOPHERUS POLYPHEMUS).

J Zoo Wildl Med 2018 Sep;49(3):520-527

Currently available tests for the diagnosis of inflammatory disease in reptiles are limited and poorly sensitive. However, a number of hematological and plasma biochemical analytes are validated in the diagnosis of inflammation in mammals. The objective of this study was to establish reference intervals for erythrocyte sedimentation rate, lactate, heat-precipitated fibrinogen, hematology, and plasma protein electrophoresis based on total protein by biuret method in 23 clinically healthy, captive gopher tortoises ( Gopherus polyphemus) after successful rehabilitation and to determine differences by age, sex, and season. In order to investigate biological differences, samples were collected in April, July, and November. There were no sex differences in any measured analyte; however, there were significant differences by age and season. Immature animals (<2 kg) had significantly higher total protein, albumin : globulin ratio, pre-albumin, albumin, and α-1 globulin than adults. Tortoises sampled in the spring season had significantly higher total solids (refractometer) and lower eosinophils compared with animals sampled in the summer. Further investigation is required to determine the clinical utility of these analytes in the diagnosis of inflammation in this species.
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http://dx.doi.org/10.1638/2017-0183.1DOI Listing
September 2018

Serum protein electrophoresis in healthy and injured southern white rhinoceros (Ceratotherium simum simum).

PLoS One 2018 25;13(7):e0200347. Epub 2018 Jul 25.

Department of Companion Animal Clinical Studies, Faculty of Veterinary Science, University of Pretoria, Pretoria, South Africa.

Investigation of globulin fractions by serum protein electrophoresis (SPE) is the first step towards evaluation of the proteome in the southern white rhinoceros (Ceratotherium simum simum). Furthermore, identification of changes in globulins in animals with poaching and other injuries can guide discovery of potentially useful biomarkers of inflammation. The aim of this study was to develop reference intervals for agarose gel SPE in healthy white rhinoceros and to compare these serum protein electrophoresis results to those from animals with tissue trauma. Reference intervals for total serum protein and agarose gel electrophoretic albumin and globulin fractions were generated using serum samples from 49 healthy free-ranging adult white rhinoceros. A standardised gating system together with identification of specific proteins by mass spectrometry aided in fraction identification. Six globulin fractions were identified: α1a, α1b, α2, β1, β2 and γ. Reference intervals were generated for total serum protein (76-111 g/L), albumin (10-27 g/L) and globulin fractions (α1a: 1.6-3.2 g/L; α1b: 1.7-3.6 g/L; α2: 16.1-26.6 g/L; β1: 6.6-18.2 g/L; β2: 11.8-30.4 g/L; γ: 10.4-23.1 g/L; albumin: globulin ratio: 0.12-0.39). Results were compared to those from 30 animals with various degrees and chronicities of tissue trauma. Wounded animals had lower concentrations of total serum protein, albumin, total globulin, α and β1 globulins, lower percentages of α2 and β1 globulins, and higher percentages of β2 and γ globulins. These protein changes are similar to those seen in human patients with wounds rather than classic acute phase or chronic inflammatory responses.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0200347PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6059428PMC
January 2019

Translational proteomic study to address host protein changes during aspergillosis.

PLoS One 2018 24;13(7):e0200843. Epub 2018 Jul 24.

University of Miami, Division of Comparative Pathology, Department of Pathology & Laboratory Medicine, Miller School of Medicine, Miami, FL, United States of America.

Aspergillosis is a fungal disease due to Aspergillus molds that can affect both humans and animals. As routine diagnosis remains difficult, improvement of basic knowledge with respect to its pathophysiology is critical to search for new biomarkers of infection and new therapeutic targets. Large-scale proteomics allows assessment of protein changes during various disease processes. In the present study, mass spectrometry iTRAQ® (isobaric tags for relative and absolute quantitation) protocol was used for direct identification and relative quantitation of host proteins in diseased fluids and tissues collected from an experimental rat model challenged with Aspergillus, as well as in blood obtained from naturally-infected penguins. In all, mass spectrometry analysis revealed that proteome during aspergillosis was mostly represented by proteins that usually express role in metabolic processes and biological process regulation. Ten and 17 proteins were significantly ≥4.0-fold overrepresented in blood of Aspergillus-diseased rats and penguins, respectively, while five and 39 were negatively ≥4.0-fold depleted within the same samples. In rat lungs, 33 proteins were identified with positive or negative relative changes versus controls and were quite different from those identified in the blood. Except for some zinc finger proteins, kinases, and histone transferases, and while three pathways were common (Wnt, cadherin and FGF), great inter-species variabilities were observed regarding the identity of the differentially-represented proteins. Thus, this finding confirmed how difficult it is to define a unique biomarker of infection. iTRAQ® protocol appears as a convenient proteomic tool that is greatly suited to ex vivo exploratory studies and should be considered as preliminary step before validation of new diagnostic markers and new therapeutic targets in humans.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0200843PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6057647PMC
January 2019

Chronic debilitation in stranded loggerhead sea turtles (Caretta caretta) in the southeastern United States: Morphometrics and clinicopathological findings.

PLoS One 2018 10;13(7):e0200355. Epub 2018 Jul 10.

Georgia Sea Turtle Center, Jekyll Island Authority, GA, United States of America.

Chronically debilitated loggerhead sea turtles (Caretta caretta) (DT) are characterized by emaciation, lethargy, and heavy barnacle coverage. Although histopathological findings associated with this condition have been reported, only limited data is available on health variables with clinical application. The objectives of this study were to 1) to compare morphometrics, clinicopathological variables, and immune functions of DTs to a group of apparently healthy loggerhead turtles to better understand the pathophysiology of the condition and 2) to assess health parameters in live debilitated turtles as they recovered during rehabilitation in order to identify potential prognostic indicators. We examined and sampled 43 DTs stranded from North Carolina to Florida for 47 health variables using standardized protocols to further characterize the condition. DTs were grouped into categories of severity of the condition, and those that survived were sampled at four time points through rehabilitation. All groups and time points were compared among DTs and to clinically healthy loggerhead turtles. Compared to healthy turtles, DTs had significantly lower body condition index, packed cell volume (PCV), total white blood cell (WBC) count, lymphocytes, glucose (Glc), total protein, all protein fractions as determined by electrophoresis, calcium (Ca), phosphorus (P), Ca:P ratio, potassium (K), lymphocyte proliferation, and greater heterophil toxicity and left-shifting, uric acid (UA), aspartate aminotransferase, creatine kinase, lysozyme, and respiratory burst. From admission to recovery, hematology and plasma chemistry data improved as expected. The most informative prognostic indicators, as determined by correlations with a novel severity indicator (based on survival times), were plastron concavity, P, albumin, total solids, UA, lymphocyte proliferation, WBC, K, Glc, Ca:P, and PCV. The results of this study document the wide range and extent of morphometric and metabolic derangements in chronically debilitated turtles. Monitoring morphometrics and clinicopathological variables of these animals is essential for diagnosis, treatment, and prognosis during rehabilitation.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0200355PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6039040PMC
January 2019