Characterization of innate immune activity in Phrynops geoffroanus (Testudines: Chelidae)

Ferronato, Bruno O.; Merchant, Mark; Marques, Thiago S.; Verdade, Luciano Martins

doi:10.1590/s1984-46702009000400020

Cited by 14 publications

(16 citation statements)

References 30 publications

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“…This pairing of physiology and behavior may optimize innate immune function during periods of maximal need and ensure a robust defense against invading pathogens. Thermal optima for plasma antibacterial activity in box turtles were higher than common snapping turtles (25–30°C), alligator snapping turtles (25–30°C), prairie rattlesnakes (25–30°C), and American alligators (15–30°C), however, they were similar to the optimum temperatures for complement‐mediated hemolysis in Geoffroy's side‐necked turtles ( Phrynops geoffroanus ; 30–40°C), European pond turtles ( Emys orbicularis ; 30–40°C), and red‐eared sliders (35°C), suggesting that phylogenetic or natural history traits may influence immune function within the Reptilia (Baker & Merchant, 2018a; Baker, Kessler, & Merchant, 2019; Ferronato, Merchant, Marques, & Verdade, 2009; Merchant et al, 2003; Yektaseresht, Gholamhosseini, & Janparvar, 2014; Yektaseresht, Gholamhosseini, & Janparvar, 2017).…”

Section: Discussionmentioning

confidence: 73%

“…However, heparin is the preferred anticoagulant for most chelonians, including box turtles, due to the occurrence of hemolysis in whole blood samples anticoagulated with EDTA (Heatley & Russell, 2010; Muro, Cuenca, Pastor, Vinas, & Lavin, 1998). Furthermore, many existing chelonian innate immune function studies (including those focused on complement activity) use plasma samples containing heparin at approximately 20 USP/ml (Baker, Kessler, Darville‐Bowleg, et al, 2019; Baker, Kessler, & Merchant, 2019; Beck et al, 2017; Cochran et al, 2018; Ferronato et al, 2009; Goessling et al, 2016; Goessling, Guyer, & Mendonça, 2017; Goessling, Koler, et al, 2017; Goessling et al, 2019; Haskins, Hamilton, Jones, et al, 2017; Haskins, Hamilton, Stacy, et al, 2017; Refsnider et al, 2015; Sandmeier et al, 2016, 2018). While our findings may be biased by the use of heparinized samples, they are comparable to the existing chelonian innate immune function literature, which also relies heavily upon this anticoagulant.…”

Section: Discussionmentioning

confidence: 99%

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Plasma antibacterial activities in ornate (Terrapene ornata) and eastern box turtles (Terrapene carolina)

et al. 2020

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Chelonians are one of the most imperiled vertebrate taxa and many species are increasingly threatened by disease, however, the immune response in this group is understudied. We quantified the innate immune response of eastern (Terrapene carolina; EBT) and ornate (Terrapene ornate; OBT) box turtles using plasma antibacterial activity assays. Plasma from both species abolished or significantly reduced the growth of all eight bacterial species evaluated, including Salmonella typhimurium, Escherichia coli, Enterobacter cloacae, Citrobacter freundi, Bacillus subtilis, Staphylococcus epidermidis, and Staphylococcus aureus. Bactericidal capacity was greater in OBT compared to EBT, and OBT plasma retained high antibacterial activities at a broader temperature range (20-40°C) compared to EBT (30-40°C). Plasma antibacterial activity was abolished following treatment with heat, protease, and ethylenediaminetetraacetic acid, indicating that complement is likely responsible for the observed effects. Further characterization of the box turtle immune response may provide insight into the importance of infectious diseases for species conservation, enabling the development of more efficient and effective population management strategies. K E Y W O R D S box turtle, chelonian, innate immunity, reptile, Terrapene

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Section: Discussionmentioning

confidence: 73%

Section: Discussionmentioning

confidence: 99%

Plasma antibacterial activities in ornate (Terrapene ornata) and eastern box turtles (Terrapene carolina)

et al. 2020

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“…The CH 50 for the American alligator (Alligator mississippiensis) and the estuarine crocodile (Crocodylus porosus) were 0.539 and 0.473 mL, respectively (Merchant & Britton, 2006). The activities for the toad-headed turtle (Phrynops geoffroanus) and the three-toed amphiuma (Amphiuma tridactylum) were so high that they could not be calculated (Ferronato et al, 2009;Major, Fontenot Jr, Pojman, & Merchant, 2011). Therefore, the comparatively low CH 50 for plasma from C. viridis indicates that it has relatively high complement activity.…”

Section: Discussionmentioning

confidence: 99%

“…Ectothermic vertebrates are typically thought to have a more effective innate immunity and less advanced adaptive immune responses (Merchant., 2013;Zimmerman, Vogel, & Bowden, 2010). Although serum complement has been studied extensively in crocodylians (Merchant et al, 2013;Merchant & Britton, 2006;Merchant, McFatter, Mead, McAdon, & Wasilewski, 2010;Merchant, Roche, Sweeney, & Elsey, 2005;Siroski, Merchant, Parachú Marcó, Pina, & Ortega, 2010), and less in turtles (Ferronato, Merchant, Marques, & Verdade, 2009) and Komodo dragons (Merchant, Henry, Falconi, Muscher, & Bryja, 2012), limited information is known concerning the details of complement activity in snakes. The few early studies that have been conducted show snakes exhibit broad-acting serum complement activities (Kawaguchi, Muramatsu, & Mitsuhashi, 1978) have an alternative pathway similar to that of mammals (Vogel & Müller-Eberhard, 1985) and produce a protein similar to that of human complement C3 (da Silva, Calich, Kipnis, & Alper, 1984).…”

Section: Introductionmentioning

confidence: 99%

Characterization of serum complement immune activity in the prairie rattlesnake (Crotalus viridis)

Baker

Merchant

2018

JoBAZ

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Background: Reptile populations face a growing number of threats including global climate change and emerging pathogens. Unfortunately, research investigating the reptile immune system lags behind other taxa groups, hindering our ability to predict or mitigate species' response to threats. Baseline studies are critical for our understanding of reptile immune response and overall health of wild populations. Results: Treatment of unsensitized sheep red blood cells (SRBCs) with plasma collected from captive prairie rattlesnakes (Crotalus viridis) resulted in volume-dependent hemolysis, with a CH 50 of 0.23 ± 0.01 mL. Kinetic analyses at different temperatures revealed that the hemolysis was relatively rapid, with 50% of hemolytic activity occurring within 15 min (25°C), 18 min (35°C), or 23 min (15°C), and near maximum hemolysis within 60-90 min at all three temperatures. A comparative thermal profile revealed that complement activity was low at 5°C, but increased sharply at 10 and 15°C, and was maximal at 20-30°C. A steep decline in activity was noted at temperatures > 30°C. Mild heat treatment of the snake plasma (56°C, 30 min) or treatment with proteases completely abolished the hemolytic activity. In addition, inclusion of 5 mM EDTA inhibited 90% of the hemolysis, but the activity could be reconstituted with the addition of 15 mM Ca 2+ or Mg 2+ , but not Ba 2+ , Cu 2+ , or Fe 2+. Furthermore, the hemolysis was unaffected in the presence of 20 mM methylamine, indicating that the alternative mechanism of complement activation is responsible for the observed activities. Conclusions: Rattlesnakes show a relatively robust innate immune response as measured by hemolysis of SRBCs. However, hemolytic activity is reduced at high temperatures, indicating that rising global temperatures may have immune consequences for snake species, making them more vulnerable to known and emerging pathogens.

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“…Distinct sensitivity to damaging agents may be an intrinsic characteristic of the species or due to adaptive mechanisms, which cannot be tested with the current data set. However, T. scripta is widespread and successful as an invasive species (Rodder et al, 2009), able to survive and reproduce in contaminated sites (Ferronato et al, 2009). The lower sensitivity to DNA damage may be a species-specific trait conferring more resilience to this species.…”

Section: Discussionmentioning

confidence: 99%

Application of the CometChip platform to assess DNA damage in field‐collected blood samples from turtles

Sýkora

Chiari

Heaton

et al. 2018

Environ and Mol Mutagen

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DNA damage has been linked to genomic instability and the progressive breakdown of cellular and organismal homeostasis, leading to the onset of disease and reduced longevity. Insults to DNA from endogenous sources include base deamination, base hydrolysis, base alkylation, and metabolism-induced oxidative damage that can lead to single-strand and double-strand DNA breaks. Alternatively, exposure to environmental pollutants, radiation or ultra-violet light, can also contribute to exogenously derived DNA damage. We previously validated a novel, high through-put approach to measure levels of DNA damage in cultured mammalian cells. This new CometChip Platform builds on the classical single cell gel electrophoresis or comet methodology used extensively in environmental toxicology and molecular biology. We asked whether the CometChip Platform could be used to measure DNA damage in samples derived from environmental field studies. To this end, we determined that nucleated erythrocytes from multiple species of turtle could be successfully evaluated in the CometChip Platform to quantify levels of DNA damage. In total, we compared levels of DNA damage in 40 animals from two species: the box turtle (Terrapene carolina) and the red-eared slider (Trachemys scripta elegans). Endogenous levels of DNA damage were identical between the two species, yet we did discover some sex-linked differences and changes in DNA damage accumulation. Based on these results, we confirm that the CometChip Platform allows for the measurement of DNA damage in a large number of samples quickly and accurately, and is particularly adaptable to environmental studies using field-collected samples. Environ. Mol. Mutagen. 59:322-333, 2018. © 2018 Wiley Periodicals, Inc.

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Characterization of innate immune activity in Phrynops geoffroanus (Testudines: Chelidae)

Cited by 14 publications

References 30 publications

Plasma antibacterial activities in ornate (Terrapene ornata) and eastern box turtles (Terrapene carolina)

Plasma antibacterial activities in ornate (Terrapene ornata) and eastern box turtles (Terrapene carolina)

Characterization of serum complement immune activity in the prairie rattlesnake (Crotalus viridis)

Application of the CometChip platform to assess DNA damage in field‐collected blood samples from turtles

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