Plasma Protein Electrophoresis Research Articles

Immune function in Trachemys scripta following exposure to a predominant brevetoxin congener, PbTx-3, as a model for potential health impacts for sea turtles naturally exposed to brevetoxins.

Many species of marine life in southwestern Florida, including sea turtles, are impacted by blooms of the toxic dinoflagellate, Karenia brevis. Sublethal exposure to toxins produced by K. brevis has been shown to impact sea turtle health. Since all sea turtles in the Gulf of Mexico have protected status, a freshwater turtle, Trachemys scripta, was used as a model for immune system effects following experimental exposure to a predominant brevetoxin congener in K. brevis blooms, PbTx-3. Exposure to PbTx-3 was oral or intratracheal and health effects were assessed using a suite of immune function parameters: innate immune function (phagocytosis, plasma lysozyme activity), adaptive immune function (lymphocyte proliferation), and measures of oxidative stress (superoxide dismutase (SOD) and glutathione-S-transferase (GST) activity in plasma). Inflammation was also measured using plasma protein electrophoresis. In addition, differential expression of genes in peripheral blood leukocytes was determined using suppression subtractive hybridization followed by real-time PCR of specific genes. The primary immune effects of sublethal brevetoxin exposure in T. scripta following PbTx-3 administration, appear to be an increase in oxidative stress, a decrease in lysozyme activity, and modulation of immune function through lymphocyte proliferation responses. Plasma protein electrophoresis showed a decreased A:G ratio which may indicate potential inflammation. Genes coding for oxidative stress, such as thioredoxin and GST, were upregulated in exposed animals. That sublethal brevetoxin exposures impact immune function components suggests potential health implications for sea turtles naturally exposed to toxins. Knowledge of physiological stressors induced by brevetoxins may contribute to the ultimate goal of developing directed treatment strategies in exposed animals for reduced mortality resulting from red tide toxin exposure in sea turtles.

Plasma protein electrophoresis in green iguanas (Iguana iguana) suffering from hepatic, renal and inflammatory skin diseases

The goals of this study were to evaluate electrophoretograms in green iguanas suffering from hepatic, renal and inflammatory skin diseases. Plasma samples were obtained from 14 green iguana patients. The diagnosis was based on clinical examination, evaluation of biochemical and haematological profiles or histopathological examination. Agarose gel electrophoresis was performed using an automated HYDRASYS system. A decrease in the albumin/globulin ratio was observed in 11 out of 14 patients (three out of four patients with liver failure, one out of one patient with chronic renal insufficiency and suspected hepatopathy, two out of two patients with chronic interstitial nephritis, four out of five patients with chronic renal insufficiency and one out of two patients with dermatitis). The alpha globulin fraction was elevated in 1 out of 14 patients (patient with dermatitis). The fraction of beta globulins was increased in 8 out of 14 patients (two out of four patients with liver diseases, four out of eight patients with various forms of renal failure and two out of two patients with inflammatory skin diseases). Increased gamma globulin levels were observed in 4 out of 14 patients (one out of four patients with impaired function of the liver, two out of eight patients with renal diseases and one out of two patients with dermatitis). Changes in the electrophoretograms of green iguanas are not disease-specific, but in conjunction with other biochemical blood tests the electrophoresis of plasma proteins may be a useful indicator in the diagnosis of metabolic and infectious diseases and in the monitoring of disease progress.

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

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.

Monitoring the health of green turtles in northern Queensland post catastrophic events

Between 2014 and 2017, the Rivers to Reef to Turtles (RRT) project examined the health of green turtles at two coastal sites impacted by urban and agricultural human activities (Cleveland and Upstart Bays) and one proposed pristine site (Howick Group of Reefs) in northern Queensland, Australia, through blood biochemistry and haematology, plasma protein electrophoresis, and clinical assessments including body condition and barnacle counts. Furthermore, cases of mortality were subjected to comprehensive postmortem examination. In an attempt to advance diagnostics, associations between specific contaminants and health of turtles in this region were tested. No comprehensive health assessments had been conducted at these sites prior to this study.The coastal Cleveland and Upstart Bays both demonstrated effects likely to be in response to stressors suspected to be anthropogenic in origin (elevated total white cell counts and creatinine kinase levels across the populations, respectively). This was associated with a suite of trace elements, in particular cobalt. While these indicators of stress resolved by the final year of the study, a chronic stressor was suspected to be persisting with ongoing low albumin: globulin. Necropsies did not elucidate any specific diseases.Although body condition index did not closely correlate with site health, barnacle counts in juvenile turtles may prove a reliable indicator of site health. Based on previously established indicators of poor health, barnacle counts showed that 10% of the population was in poor health at Upstart Bay and nearly 20% of the population at Cleveland Bay. This is above what would be expected for a normal population.Overall, the health component of this study suggested that the pristine turtle population was healthy and the coastal turtle populations were under active stressors, possibly caused by anthropogenic effectors such as chemical pollutants, when initially examined in 2014. These stressors resolved by the conclusion of the study in 2017; but chronic stressors remained absent in the pristine site and present within each of the studied coastal populations.

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

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.

Serum proteins in healthy and diseased Florida manatees (Trichechus manatus latirostris)

A major goal of this study was to determine whether serum protein fractions of healthy Florida manatees differ with age, sex, or living environments (wild versus housed). A second goal was to determine which serum protein fractions vary in diseased versus healthy manatees. Serum protein fractions were determined using agarose gel electrophoresis. Healthy adults had slightly higher total serum protein and total globulin concentrations than younger animals. This largely resulted from an increase in gamma globulins with age. Total serum protein, albumin, alpha-1 globulin, beta globulin, and total globulin concentrations were slightly higher in housed manatees compared to wild manatees, but there was no significant difference in the albumin/globulin (A/G) ratio, suggesting a difference in hydration between these groups. No significant differences were attributable to sex or pregnancy. Serum albumin concentrations and A/G ratios were significantly lower for manatees with boat trauma, entanglements, emaciation, or cold stress compared to healthy manatees. Variable increases were seen in alpha-1globulins, alpha-2 globulins, beta globulins, and gamma globulins. These globulin fractions contain positive acute-phase proteins and immunoglobulins, and their increases may reflect acute or chronic active inflammation. Changes in serum protein fractions were not consistent enough to justify the use of serum protein electrophoresis as a routine diagnostic test for manatees. However, serum (or plasma) protein electrophoresis is required when accurate values for albumin and globulins are needed in manatees and in determining which protein fractions may account for a hyperproteinemia or hypoproteinemia reported in a clinical chemistry panel.

EVALUATION OF HEALTH PARAMETERS IN COWNOSE RAYS ( RHINOPTERA BONASUS) HOUSED IN A SEASONAL TOUCH POOL HABITAT COMPARED WITH AN OFF-EXHIBIT HABITAT.

Cownose rays ( Rhinoptera bonasus) are commonly displayed in zoo and aquarium touch pool exhibits; however, there is a gap in our understanding of how these practices might impact the health of these animals. The aim of this study was to evaluate and compare selected health parameters in cownose rays housed in a seasonal outdoor exhibit touch pool system with abundant public contact and an indoor off-exhibit holding system with minimal human contact. All animals underwent physical examination, ultrasound, cloacal wash and cytology, and blood collection for complete blood counts, point-of-care blood analysis, plasma protein electrophoresis, and plasma cholesterol electrophoresis in May and October 2014. Physical examination, ultrasound, and cloacal wash cytology findings were all unremarkable for both groups of animals. Significant differences in health parameters among animals by location and time point were few and included decreased heart rate ( F = 12.158, P = 0.001), increased lactate ( F = 6.838, P = 0.012), and increased low-density lipoproteins ( F = 19.961, P = 0.000) in touch pool animals over time. Based on these results, cownose rays inhabiting a touch pool exhibit and an off-exhibit system remained in comparable planes of health based on routine diagnostic modalities with few differences in measured health parameters.

Testing of an oral dosing technique for double-crested cormorants, Phalacocorax auritus, laughing gulls, Leucophaeus atricilla, homing pigeons, Columba livia, and western sandpipers, Calidris mauri, with artificially weather MC252 oil

Scoping studies were designed to determine if double-crested cormorants (Phalacocorax auritus), laughing gulls (Leucophaues atricilla), homing pigeons (Columba livia) and western sandpipers (Calidris mauri) that were gavaged with a mixture of artificially weathered MC252 oil and food for either a single day or 4–5 consecutive days showed signs of oil toxicity. Where volume allowed, samples were collected for hematology, plasma protein electrophoresis, clinical chemistry and electrolytes, oxidative stress and organ weigh changes. Double-crested cormorants, laughing gulls and western sandpipers all excreted oil within 30min of dose, while pigeons regurgitated within less than one hour of dosing. There were species differences in the effectiveness of the dosing technique, with double-crested cormorants having the greatest number of responsive endpoints at the completion of the trial. Statistically significant changes in packed cell volume, white cell counts, alkaline phosphatase, alanine aminotransferase, creatine phosphokinase, gamma glutamyl transferase, uric acid, chloride, sodium, potassium, calcium, total glutathione, glutathione disulfide, reduced glutathione, spleen and liver weights were measured in double-crested cormorants. Homing pigeons had statistically significant changes in creatine phosphokinase, total glutathione, glutathione disulfide, reduced glutathione and Trolox equivalents. Laughing gulls exhibited statistically significant decreases in spleen and kidney weight, and no changes were observed in any measurement endpoints tested in western sandpipers.

Plasma protein, haematologic and blood chemistry changes in African grey parrots (Psittacus erithacus) experimentally infected with bornavirus

ABSTRACTBornaviruses are considered to be the causative agent of proventricular dilatation disease (PDD) in psittacine birds. In order to detect haematological and blood chemistry changes during the development of PDD and a possible correlation with clinical signs and the virological status, six African grey parrots (Psittacus erithacus) were experimentally infected with parrot bornavirus 4 (PaBV-4) by subcutaneous route. All six parrots developed clinical signs of varying extent and successful infection was confirmed in all the birds by seroconversion or detection of RNA of the PaBV-4 infection strain. Based on population-based and intra-individual reference ranges established during 12 months prior to experimental infection, only minor haematological changes were detected in individual birds after infection. Changes in blood chemistry were restricted to aspartate aminotransferase, creatine kinase, total protein, glucose and uric acid. Plasma protein electrophoresis revealed marked changes starting 10 weeks post infection characterized by an increase in the γ-globulin fraction and a gradual decrease to normal values during weeks 22–34. Indications of an acute-phase reaction at the initial stages of infection were not detected. While three birds suffered from clinical signs of PDD, which included weight loss and neurological disorders and died before development of haematological and plasma protein changes, recovery of clinical disease was paralleled in the remaining birds by an increase in γ-globulins and bornavirus-specific antibody titres.

Potential effects of brevetoxins and toxic elements on various health variables in Kemp's ridley (Lepidochelys kempii) and green (Chelonia mydas) sea turtles after a red tide bloom event

Natural biotoxins and anthropogenic toxicants pose a significant risk to sea turtle health. Documented effects of contaminants include potential disease progression and adverse impacts on development, immune function, and survival in these imperiled species. The shallow seagrass habitats of Florida's northwest coast (Big Bend) serve as an important developmental habitat for Kemp's ridley (Lepidochelys kempii) and green (Chelonia mydas) sea turtles; however, few studies have been conducted in this area. Our objectives were (1) to evaluate plasma analytes (mass, minimum straight carapace length, body condition index [BCI], fibropapilloma tumor score, lysozyme, superoxide dismutase, reactive oxygen/nitrogen species, plasma protein electrophoresis, cholesterol, and total solids) in Kemp's ridleys and green turtles and their correlation to brevetoxins that were released from a red tide bloom event from July–October 2014 in the Gulf of Mexico near Florida's Big Bend, and (2) to analyze red blood cells in Kemp's ridleys and green turtles for toxic elements (arsenic, cadmium, lead, mercury, selenium, thallium) with correlation to the measured plasma analytes. Positive correlations were observed between brevetoxins and α2-globulins in Kemp's ridleys and α2- and γ-globulins in green turtles, indicating potential immunostimulation. Arsenic, cadmium, and lead positively correlated with superoxide dismutase in Kemp's ridleys, suggesting oxidative stress. Lead and mercury in green turtles negatively correlated with BCI, while mercury positively correlated with total tumor score of green turtles afflicted with fibropapillomatosis, suggesting a possible association with mercury and increased tumor growth. The total tumor score of green turtles positively correlated with total protein, total globulins, α2-globulins, and γ-globulins, further suggesting inflammation and immunomodulation as a result of fibropapillomatosis. Lastly, brevetoxin concentrations were positively related to tumor score, indicating potential tumor promotion by brevetoxin. These results signify that brevetoxins and toxic elements elicit various negative effects on sea turtle health, including immune function, oxidative stress, and possibly disease progression.

STRESS INFLUENCE ON PLASMA PROTEIN ELECTROPHORESIS IN THE RED-BREASTED GOOSE (BRANTA RUFICOLLIS) AND HAWAIIAN GOOSE (BRANTA SANDVICENSIS).

The objective of this study was to evaluate the effects of stress (as measured by total heterophil + eosinophil counts [THECs] and plasma corticosterone [PC] levels) on plasma protein electrophoresis (PPE) in two Anseriform species. Ten red-breasted geese ( Branta ruficollis ) and eight Hawaiian geese (Branta sandvicensis) were received into quarantine at Beauval Zoo and housed together with other Anseriformes. Two days later, all were examined as part of routine quarantine procedures, and blood was collected. THECs were performed using a Malassez hemocytometer after a 1 : 200 dilution with an eosinophil dilution liquid containing phloxine B. PPE, as well as total protein and PC assays, was performed on lithium heparin plasma. Twenty days after arrival, all birds were similarly restrained, examined, and sampled. Pododermatitis lesions were identified on four geese that were excluded from statistical analysis. For each goose, THECs and PC values were sorted by value (lower or higher) independently from the day of sampling. A Wilcoxon signed rank test showed no significant differences between lower and higher values of THECs for any of the PPE fractions. Higher values of corticosterone were associated with higher values of prealbumin, but none of the other fractions were significantly different. A Spearman rank correlation coefficient showed that THECs and PC were not correlated; this suggested differences in kinetics between these stress markers. Results did not show significant alterations of electrophoresis patterns associated with stress as measured with THECs and PC. The results also highlight the complexity of precisely assessing acute and chronic stress in avian species.

CKS1B/CDKN2C (P18) amplification/deletion as prognostic markers in multiple myeloma patients

Introduction Multiple myeloma (MM) is neoplasia of plasma cells characterized by clonal proliferation of malignant plasma cells in the bone marrow. Amplification of 1q21 (CKS1B) is the most common recurrent chromosomal aberration in myeloma. Overexpression of the CKS1B gene upregulates cell cycle processes and leads to a more proliferative malignant plasma cell. This is related to an unfavorable clinical course with poor prognosis and disease progression. Also the deletions of CDKN2C have been identified in 40% of the studied MM cases and hence inactivation of CDKN2C may be an important step in the initiation and progression of MM and could be an important prognostic factor. The aim of the work The aim of the study was to assess the prognostic significance of CKS1B gain and CDKN2C gene deletion in MM patients. Patients and methods This prospective cohort study was carried out on 40 newly diagnosed MM patients who were submitted to routine laboratory investigations, including complete blood count, bone marrow aspiration, blood urea nitrogen, creatinine, calcium, albumin, plasma protein electrophoresis, 24 h urine for total protein, and β2-microglobulin, and to detection of CKS1B/CDKN2C (P18) amplification/deletion by fluorescence in-situ hybridization technique. Results On using fluorescence in-situ hybridization, we found that 25% (10 out of 40) of patients showed CKS1B gain and 20% (eight out of 40) showed positive deletion of the CDKN2C gene. All these patients showed inferior outcome and short survival. Conclusion MM patients with CKS1B/CDKN2C (P18) amplification/deletion had a more aggressive disease with adverse impact on survival, which makes CKS1B and CDKN2C genes valuable prognostic indicators in MM patients.

Acquired and congenital fast albumin bands; insights from electrospray TOF analysis of whole plasma into drug binding and albumin recycling

ObjectivesTo define the underlying cause of unusual fast albumin bands detected on plasma protein electrophoresis of two patients. MethodPlasma was examined by electrospray time-of-flight mass spectrometry (TOF MS) to assess the possibility of congenital or acquired structural modifications. ResultsIn one patient whole protein MS indicated a drop of 486Da in the mass of 5.1% of the albumin molecules. This and the presence of an additional minor product (65,806Da) lacking a C-terminal phenylalanine (−147Da) indicated that this was albumin Rugby Park; an electrophoretically fast albumin variant caused by a splice site mutation (GT>CT) in intron 13 of the albumin gene. The second patient had an acquired alteration with a drift of albumin mobility to the anode. This severely ill patient was on intra venous antibiotics and electrospray TOF MS showed a stuttered repetition of the 66,439/66,558Da albumin isoforms at multiples of 455–459Da corresponding to the covalent attachment of 1, 2, 3 and 4 molecules of flucloxacillin. This modification of +455Da was also detected in a control on a 1g/day oral dose of flucloxacillin. ConclusionBoth aberrations were associated with diminished albumin concentrations. The C-terminal truncation of Rugby Park (albumin, 29g/l) likely interferes with receptor binding and albumin scavenging, while the 20g/l albumin in the second patient was mostly due to renal disease. In both cases electrospray TOF MS proved a rapid (5min) sensitive (0.2μl plasma) and highly informative way of analysing whole plasma or serum.

Variations of plasma protein electrophoresis in healthy captive Green Iguanas (Iguana iguana).

Serum or plasma protein electrophoresis is used as a routine test for health assessment in veterinary medicine, but there are only a limited number of studies regarding clinical use of electrophoresis in reptile species. The goals of this study were to establish reference intervals for plasma protein electrophoresis in the Green Iguana (Iguana iguana), compare values between males and females, and to identify season-related changes. Plasma samples were obtained from 21 healthy captive male and female Green Iguanas. Agarose gel electrophoresis was performed using an automated Hydrasys system. Four main protein fractions were observed: albumin, α globulins, β globulins, and γ globulins. Bisalbuminemia was observed in 4 of 21 healthy iguanas. Minimum and maximum values were reported for healthy Green Iguanas in March, June, September, and December. Seasonal changes in albumin were determined between March and December, and in γ globulins between June and September. Differences between males and females were seen in albumin concentration in September. Reference intervals of the plasma protein fractions according to electrophoresis in the Green Iguana can be affected by seasonal changes and sex of animals. It should be taken into account when clinical evaluation is performed.

Sublethal red tide toxin exposure in free-ranging manatees (Trichechus manatus) affects the immune system through reduced lymphocyte proliferation responses, inflammation, and oxidative stress

The health of many Florida manatees (Trichechus manatus latirostris) is adversely affected by exposure to blooms of the toxic dinoflagellate, Karenia brevis. K. brevis blooms are common in manatee habitats of Florida’s southwestern coast and produce a group of cyclic polyether toxins collectively referred to as red tide toxins, or brevetoxins. Although a large number of manatees exposed to significant levels of red tide toxins die, several manatees are rescued from sublethal exposure and are successfully treated and returned to the wild. Sublethal brevetoxin exposure may potentially impact the manatee immune system. Lymphocyte proliferative responses and a suite of immune function parameters in the plasma were used to evaluate effects of brevetoxin exposure on health of manatees rescued from natural exposure to red tide toxins in their habitat. Blood samples were collected from rescued manatees at Lowry Park Zoo in Tampa, FL and from healthy, unexposed manatees in Crystal River, FL. Peripheral blood leukocytes (PBL) isolated from whole blood were stimulated with T-cell mitogens, ConA and PHA. A suite of plasma parameters, including plasma protein electrophoresis profiles, lysozyme activity, superoxide dismutase (SOD) activity, and reactive oxygen/nitrogen (ROS/RNS) species, was also used to assess manatee health. Significant decreases (p<0.05) in lymphocyte proliferation were observed in ConA and PHA stimulated lymphocytes from rescued animals compared to non-exposed animals. Significant correlations were observed between oxidative stress markers (SOD, ROS/RNS) and plasma brevetoxin concentrations. Sublethal exposure to brevetoxins in the wild impacts some immune function components, and thus, overall health, in the Florida manatee.

Establishment of reference intervals for plasma protein electrophoresis in Indo-Pacific green sea turtles, Chelonia mydas.

Biochemical and haematological parameters are increasingly used to diagnose disease in green sea turtles. Specific clinical pathology tools, such as plasma protein electrophoresis analysis, are now being used more frequently to improve our ability to diagnose disease in the live animal. Plasma protein reference intervals were calculated from 55 clinically healthy green sea turtles using pulsed field electrophoresis to determine pre-albumin, albumin, α-, β- and γ-globulin concentrations. The estimated reference intervals were then compared with data profiles from clinically unhealthy turtles admitted to a local wildlife hospital to assess the validity of the derived intervals and identify the clinically useful plasma protein fractions. Eighty-six per cent {19 of 22 [95% confidence interval (CI) 65-97]} of clinically unhealthy turtles had values outside the derived reference intervals, including the following: total protein [six of 22 turtles or 27% (95% CI 11-50%)], pre-albumin [two of five, 40% (95% CI 5-85%)], albumin [13 of 22, 59% (95% CI 36-79%)], total albumin [13 of 22, 59% (95% CI 36-79%)], α- [10 of 22, 45% (95% CI 24-68%)], β- [two of 10, 20% (95% CI 3-56%)], γ- [one of 10, 10% (95% CI 0.3-45%)] and β-γ-globulin [one of 12, 8% (95% CI 0.2-38%)] and total globulin [five of 22, 23% (8-45%)]. Plasma protein electrophoresis shows promise as an accurate adjunct tool to identify a disease state in marine turtles. This study presents the first reference interval for plasma protein electrophoresis in the Indo-Pacific green sea turtle.

Plasma proteins and selected acute-phase proteins in the whitespotted bamboo shark (Chiloscyllium plagiosum).

Plasma protein electrophoresis has been shown to be an important tool when recognizing disease in various species. Plasma was harvested from twenty captive whitespotted bamboo sharks (Chiloscyllium plagiosum) and gel electrophoresis was used to characterize the following protein fractions: total protein, prealbumin, albumin, α-1 globulin, α-2 globulin, β globulin, γ globulin, C-reactive protein, serum amyloid A, and haptoglobin. The selected acute-phase proteins were assayed and reference intervals created for a population of captive whitespotted bamboo sharks, and then used to assess relationships between these protein concentration values, sex, and health status. There were significantly higher β fractions and total protein in females than in males. There did not appear to be a relationship between physical examination abnormalities of individual sharks and their plasma protein levels. Further research is needed to characterize the clinical importance of this response.

Plasma Protein Electrophoresis and Acute Phase Proteins in Koi Carp (Cyprinus carpio) Following Exploratory Coeliotomy

Measuring plasma protein electrophoresis fractions and plasma concentrations of acute phase proteins (APPs) is commonly used to monitor inflammation and illness in mammalian species, but its use is not widespread in fish species, despite the presence of similar proteins. This study aims to determine plasma protein electrophoresis values for koi carp (Cyprinus carpio) and to assess the use of plasma protein electrophoresis and commercially available assays for 3 primary APPs (C-reactive protein, serum amyloid A, and haptoglobin) as markers of inflammation in 13 koi following invasive surgery. Median packed cell volume decreased by nearly 50% but recovered to baseline by 14 days postsurgery. Plasma protein electrophoresis fractions were consistent across the experimental period and did not demonstrate an inflammatory pattern, but electrophoretograms exhibited variation between individual fish. No clinically meaningful changes were observed in the APPs of the test subjects 28 days following surgery. Despite grossly evident inflammation of the incision line, either this surgical procedure did not stimulate a marked systemic inflammatory response, or the available assays are not effective in detecting the acute phase inflammatory response in this fish species.

Influence of short-term storage conditions on the stability of total protein concentrations and electrophoretic fractions in plasma samples from loggerhead sea turtles, Caretta caretta

Plasma protein electrophoresis has been proposed for use in reptile medicine to aid in the diagnosis of disease. Reference intervals of plasma proteins in loggerhead sea turtles have been established; however, studies evaluating the effect of storage conditions on plasma protein and protein fractions in this species are lacking. The aim of this study was to evaluate the effect of short-term storage conditions on total protein concentration and electrophoretic fractions of sea turtle plasma. Blood was collected from sea turtles, and plasma obtained after centrifugation was fractioned into five aliquots. One aliquot was analyzed within 3 h after collection, whereas the remaining aliquots were stored at either +4 °C or −20 °C for 2 or 7 days. Overall, there was a significant effect of storage temperature and duration on albumin, β-globulin, and A/G ratio (Friedman rank sum test P < 0.001) but not on total protein concentration. The results of the current study show that short-term storage conditions influence the electrophoretic pattern of plasma proteins and the A/G ratio in Caretta caretta. Therefore, in order to obtain plasma protein concentrations that can serve as reliable screening tests in veterinary diagnostic laboratories, practitioners should refrigerate plasma for no more than 2 days.

Assessment of hemoglobin binding protein in loggerhead sea turtles (Caretta caretta) undergoing rehabilitation.

The acute phase response is an important component of the early reaction of the immune system to insults including infection, inflammation, trauma, neoplasia, and stress. Acute phase proteins are valuable prognostic indicators in many mammalian species but have been poorly studied in reptiles thus far. This study examined 18 paired samples from loggerhead sea turtles (Caretta caretta) for changes observed during the rehabilitation period. Analyses performed included packed cell volume (PCV), hemoglobin binding protein, and plasma protein electrophoresis. Significant differences were observed in all of the protein electrophoresis values. Notably, the concentration of hemoglobin binding protein (as determined by a haptoglobin assay) increased in conjunction with rising total protein (by refractometry) and PCV. The results indicate that this assay may have the potential to be a useful tool in assessing the health of sea turtles.