Carolyn Cray

Comparison of Blood Values in Foraging, Nesting, and Stranded Loggerhead Turtles (Caretta Caretta) Along the Coast of Georgia, Usa

The health status of 83 loggerhead sea turtles (Caretta caretta; 39 foraging, 31 nesting, and 13 stranded turtles) was analyzed using physical examinations, hematology, plasma biochemistry, plasma protein electrophoresis, and toxicologic parameters. Significant differences were noted in a number of health parameters between turtles exhibiting each of these behaviors. On physical examinations, stranded turtles had the highest prevalence of heavy carapace epibiont loads, miscellaneous abnormalities, emaciation, and weakness. Differences in hematologic values included a lower packed cell volume, higher number of lymphocytes, and lower number of monocytes in stranded turtles; lower white blood cell counts in foraging turtles; and significant differences in total solid values among turtles exhibiting all behaviors with the lowest values in stranded turtles and the highest values in nesting turtles. Differences in plasma biochemistry values included the highest uric acid, creatine kinase, and CO2 values in stranded turtles; the highest glucose and potassium values in foraging turtles; and the highest cholesterol and triglyceride values, and lowest alanine aminotransferase, in nesting turtles. Differences in total protein, albumin, and globulin were found using plasma biochemistry values, with lowest values in stranded turtles and highest values in nesting females, whereas differences in blood urea nitrogen between turtles included the lowest values in nesting turtles and the highest in foraging turtles. Plasma organochlorine and polychlorinated biphenyl levels were below their limits of quantification in the 39 foraging, 11 nesting, and three stranded turtles tested. A statistically significant difference was noted in the level of whole blood mercury between the 23 foraging and 12 nesting turtles tested. There was no difference in arsenic or lead levels between turtles exhibiting any of the three behaviors. Although a few limitations exist with the present study and include unknown ambient temperatures, turtle handling times that varied from 15 min to 53 min per turtle, and the use of a different laboratory for processing complete blood counts and plasma biochemistries in stranded versus foraging and nesting turtles, we provide baseline blood values for two cohorts (foraging and nesting) of loggerhead sea turtles on the coast of Georgia. Additionally, we demonstrate significant differences in clinical findings and blood parameters between foraging, nesting, and stranded loggerhead turtles in the region.

Acute Phase Proteins in Animals

Acute phase proteins (APP) were first identified in the early 1900s as early reactants to infectious disease. They are now understood to be an integral part of the acute phase response (APR) which is the cornerstone of innate immunity. APP have been shown to be valuable biomarkers as increases can occur with inflammation, infection, neoplasia, stress, and trauma. All animals—from fish to mammals—have demonstrable APP, but the type of major APP differs by species. While the primary application of these proteins in a clinical setting is prognostication, studies in animals have demonstrated relevance to diagnosis and detection and monitoring for subclinical disease. APP have been well documented in laboratory, companion, and large animals. With the advent of standardized and automated assays, these biomarkers are available for use in all fields of veterinary medicine as well as basic and clinical research.

IMMUNE STATUS OF FREE-RANGING GREEN TURTLES WITH FIBROPAPILLOMATOSIS FROM HAWAII

Cell-mediated and humoral immune status of free-ranging green turtles (Chelonia mydas) in Hawaii (USA) with and without fibropapillomatosis (FP) were assessed. Tumored and non-tumored turtles from Kaneohe Bay (KB) on the island of Oahu and from FP-free areas on the west (Kona/Kohala) coast of the island of Hawaii were sampled from April 1998 through February 1999. Turtles on Oahu were grouped (0–3) for severity of tumors with 0 for absence of tumors, 1 for light, 2 for moderate, and 3 for most severe. Turtles were weighed, straight carapace length measured and the regression slope of weight to straight carapace length compared between groups (KB0, KB1, KB2, KB3, Kona). Blood was assayed for differential white blood cell count, hematocrit, in vitro peripheral blood mononuclear cell (PBMC) proliferation in the presence of concanavalin A (ConA) and phytohaemagglutinin (PHA), and protein electrophoresis. On Oahu, heterophil/lymphocyte ratio increased while eosinophil/monocyte ratio decreased with increasing tumors score. Peripheral blood mononuclear cell proliferation indices for ConA and PHA were significantly lower for turtles with tumor scores 2 and 3. Tumor score 3 turtles (KB3) had significantly lower hematocrit, total protein, alpha 1, alpha 2, and gamma globulins than the other four groups. No significant differences in immune status were seen between non-tumored (or KB1) turtles from Oahu and Hawaii. There was no significant difference between groups in regression slopes of body condition to carapace length. We conclude that turtles with severe FP are imunosuppressed. Furthermore, the lack of significant difference in immune status between non-tumored (and KB1) turtles from Oahu and Kona/Kohala indicates that immunosuppression may not be a prerequisite for development of FP.

Restricted Enzooticity of Hepatitis E Virus Genotypes 1 to 4 in the United States

ABSTRACT Hepatitis E is recognized as a zoonosis, and swine are known reservoirs, but how broadly enzootic its causative agent, hepatitis E virus (HEV), is remains controversial. To determine the prevalence of HEV infection in animals, a serological assay with capability to detect anti-HEV-antibody across a wide variety of animal species was devised. Recombinant antigens comprising truncated capsid proteins generated from HEV-subgenomic constructs that represent all four viral genotypes were used to capture anti-HEV in the test sample and as an analyte reporter. To facilitate development and validation of the assay, serum samples were assembled from blood donors (n = 372), acute hepatitis E patients (n = 94), five laboratory animals (rhesus monkey, pig, New Zealand rabbit, Wistar rat, and BALB/c mouse) immunized with HEV antigens, and four pigs experimentally infected with HEV. The assay was then applied to 4,936 sera collected from 35 genera of animals that were wild, feral, domesticated, or otherwise held captive in the United States. Test positivity was determined in 457 samples (9.3%). These originated from: bison (3/65, 4.6%), cattle (174/1,156, 15%), dogs (2/212, 0.9%), Norway rats (2/318, 0.6%), farmed swine (267/648, 41.2%), and feral swine (9/306, 2.9%). Only the porcine samples yielded the highest reactivities. HEV RNA was amplified from one farmed pig and two feral pigs and characterized by nucleotide sequencing to belong to genotype 3. HEV infected farmed swine primarily, and the role of other animals as reservoirs of its zoonotic spread appears to be limited.

Assessment of serum amyloid A testing of horses and its clinical application in a specialized equine practice

OBJECTIVE To compare serum amyloid A (SAA) concentration, plasma fibrinogen concentration, total WBC count, and serum albumin-to-globulin concentration ratio (A:G ratio) in clinically normal (CN) and clinically abnormal (CA) horses. DESIGN Prospective cohort study. ANIMALS 111 CN horses and 101 CA horses hospitalized at a specialty clinical practice. PROCEDURES Shortly after admission, a blood sample (20 mL) was collected from each horse for a CBC, serum protein electrophoresis, and determination of plasma fibrinogen concentration; SAA concentration was assessed with a previously validated immunoturbidometric assay. Similar testing of a subset of CA horses was conducted at various points during treatment. RESULTS Total WBC count, A:G ratio, and SAA concentration were determined for all 212 horses; data regarding plasma fibrinogen concentration were available for 127 horses (of which 47 were CN and 80 were CA). Median SAA concentration, total WBC count, and plasma fibrinogen concentration and mean A:G ratio differed significantly between CN horses and CA horses. Correlations between these variables were poor to weak. For discrimination of CN horses from CA horses, the SAA assay had sensitivity of 53% and specificity of 94% (diagnostic accuracy, 75%); for the other assessments, accuracy ranged from 59% to 62%. Repeated assessment of SAA concentration in some CA horses revealed a gradual return to normal concentrations. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that assessment of SAA concentration can provide valuable information regarding the clinical state of horses and may be more useful for patient monitoring and as a prognostic indicator than are traditional markers of inflammation.

ALTERED IN VITRO IMMUNE RESPONSES IN GREEN TURTLES (CHELONIA MYDAS) WITH FIBROPAPILLOMATOSIS

Abstract The immune competence of green sea turtles (Chelonia mydas) with fibropapillomatosis was assessed using in vitro techniques to measure lymphocyte proliferation in response to mitogens. In comparison with captive, healthy green sea turtles, those afflicted with fibropapillomas demonstrated diminished proliferation with Concanavalin A, phytohemagglutinin (T-cell mitogens), and lipopolysaccharide (B-cell mitogen). Also, markedly decreased proliferative responses to the lymphocyte polyclonal stimulator combination of ionomycin and phorbol myristate acetate were observed. Total circulating white blood cell counts were not statistically different between the two groups, although an overall decrease in lymphocyte number was observed in the papilloma group. The albumin/globulin ratio was decreased in the papilloma group because of decreased albumin and increased gamma globulins.

Evaluation of the usefulness of an ELISA and protein electrophoresis in the diagnosis of Encephalitozoon cuniculi infection in rabbits

OBJECTIVE-To evaluate the usefulness of an antibody detection ELISA and protein electrophoresis (PE) for diagnosing Encephalitozoon cuniculi (ECUN) infection in pet rabbits. ANIMALS-203 pet rabbits. PROCEDURES-Serum and plasma samples from pet rabbits were submitted from veterinary clinics within the United States. Participating veterinarians completed a questionnaire that was used to classify rabbits as clinically normal (n=33), suspected of having an ECUN infection (103), or clinically abnormal but not suspected of having an ECUN infection (67). An ELISA for detection of serum or plasma IgG against ECUN was developed by use of commercially available reagents. Results of the ELISA and PE were used to detect ECUN infection. RESULTS-A high seroprevalence of antibody against ECUN was detected in all 3 groups of rabbits. In rabbits suspected of having an ECUN infection, the mean IgG titer was 1.7 times as high as the values in the other rabbit groups. Rabbits suspected of having an ECUN infection and those that were simply clinically abnormal had a higher concentration of gamma-globulins than clinically normal rabbits. This increase in globulins concentration was accompanied by a decrease in the albumin-to-globulin ratio. Results of the ELISA and PE were significantly different between clinically normal rabbits and those suspected of having an ECUN infection. CONCLUSIONS AND CLINICAL RELEVANCE-The combination of an ELISA and PE may aid in the diagnosis of ECUN infection in pet rabbits. IMPACT FOR HUMAN MEDICINE-Because ECUN is a potential zoonotic agent, diagnostic methods for pet rabbits need to be improved to protect human health.

Protein Electrophoresis: A Tool for the Reptilian and Amphibian Practitioner

ABSTRACT This review is intended to introduce reptilian and amphibian practitioners to the practice and benefits of protein electrophoresis as a supplemental diagnostic tool. Protein electrophoresi...

Galactomannan Assay and Plasma Protein Electrophoresis Findings in Psittacine Birds With Aspergillosis

Abstract In psittacine birds, the antemortem diagnosis of aspergillosis is usually based on the clinical signalment combined with the results of diagnostic tests such as radiography, routine hematologic and biochemical analysis, and biopsy. For several years, plasma protein electrophoresis has been used as an ancillary diagnostic technique in forming a diagnosis and treatment plan in avian species. More recently, a commercially available assay to measure galactomannan, an Aspergillus species antigen, has been described for clinical use in humans, cattle, horses, dogs, and gyr falcons. This report describes several confirmed cases of aspergillosis, with accompanying clinical data, including plasma protein electrophoresis and galactomannan assay results, in addition to results of traditional evaluations by hematology, radiography, and biopsy. In clinical cases in psittacine birds, the galactomannan assay appears useful for detecting circulating Aspergillus antibody.

Serosurvey and Diagnostic Application of Antibody Titers to Aspergillus in Avian Species

Abstract A multiyear study was conducted using an enzyme-linked immunosorbent assay to measure antibody to address the application of the test to the diagnosis of aspergillosis in avian species. In general serostudies (n  =  1314), four avian groups (psittaciform, raptor, penguin, and zoo) were found to have samples with antibody reactivity. Penguin, raptor, and zoo groups were found to have higher levels of antibody to Aspergillus than the psittaciform group. Additional clinical information was collected on 303 cases, which resulted in the definition of presumptive normal, probable, and confirmed infection groups. Although the confirmed group was more likely to have antibody reactivity, the mean antibody index was not found to be significant between presumptive normal and probable or confirmed cases.