Is There Good Simulation Basic Training for End-To-Side Vascular Microanastomoses?

Background: Microvascular anastomosis is the cornerstone of free tissue transfers. Irrespective of the microsurgical technique that one seeks to integrate or improve, the time commitment in the laboratory is significant. After extensive previous training on several animal models, we sought to identify an animal model that circumvents the following issues: ethical rules, cost, time-consuming and expensive anesthesia, and surgical preparation of tissues required to access vessels before performing the microsurgical training, not to mention that laboratories are closed on weekends. Methods: Between January 2012 and April 2012, a total of 91 earthworms were used for 150 microsurgical training exercises to simulate vascular end-to-side microanastomosis. The training sessions were divided into ten periods of 7days. Each training session included 15 simulations of end-to-side vascular microanastomoses: larger than 1.5mm (n=5), between 1.0 and 1.5mm (n=5), and smaller than 1.0mm (n=5). A linear model with the main variables being the number of weeks (as a numerical covariate) and the size of the animal (as a factor) was used to determine the trend in time of anastomosis over subsequent weeks as well as the differences between the different size groups. Results: The linear model shows a significant trend (p<0.001) in time of anastomosis in the course of the training, as well as significant differences (p<0.001) between the groups of animals of different sizes. For microanastomoses larger than 1.5mm, the mean anastomosis time decreased from 19.3±1.0 to 11.1±0.4min between the first and last week of training (decrease of 42.5%). For training with smaller diameters, the results showed a decrease in execution time of 43.2% (diameter between 1.0 and 1.5mm) and 40.9% (diameter <1.0mm) between the first and last periods. The study demonstrates an improvement in the dexterity and speed of nodes execution. Conclusion: The earthworm appears to be a reliable experimental model for microsurgical training of end-to-side microanastomoses. Its numerous advantages are discussed here and we predict training on earthworms will significantly grow and develop in the near future. Level of Evidence III: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/0026

First evidence of recruitment in critically endangered Galápagos pink land iguanas (Conolophus marthae)

We present the first estimate of population dynamics parameters for critically endangered Galápagos pink land iguanas, Conolophus marthae, leveraging mark-recapture data collected between 2006 and 2021. Demographic parameters were estimated using the POPAN formulation of the Jolly–Seber model. This approach provided a solid and robust estimate, consolidating the results obtained from previous estimates based on the Lincoln–Petersen method evaluating only two sampling occasions. Our results suggest that the abundance of males was higher than that of females at each sampling occasion, probably due to differences in sex-specific behavior during the reproductive season. Our analyses also provide no evidence of a change in population size during the sampling period, estimated at 150–270 adult individuals, while suggesting positive entry rates. Finally, by comparing the biometric features of first-captured versus recaptured iguanas, we found that the former have more juvenile-like characteristics (i.e., they are, on average, smaller) than the latter. Although juvenile pink iguanas have rarely been recorded (only four sampled juvenile-like individuals in over 16 years of field work), our combined results provide the first clear indication that the only known C. marthae population actively recruits new members from younger age classes. Such recruitment may have prevented any measurable population decline in the last 16 years, but it was also not sufficient to clearly increase the population size. These results are of the utmost importance for the conservation of this species. They will guide future high-priority actions: (i) aiming at increasing population size by removing or mitigating threats and (ii) increasing the number of individuals by implementing a headstart program and/or founding a second viable population, as identified in the recently published IUCN Conservation and Management Plan

Green algal infection of American horseshoe crab (Limulus polyphemus) exoskeletal structures

Author Posting. © The Author(s), 2012. This is the author's version of the work. It is posted here by permission of Elsevier B.V. for personal use, not for redistribution. The definitive version was published in Journal of Invertebrate Pathology 111 (2012): 90-93, doi:10.1016/j.jip.2012.06.002.Degenerative lesions in the dorsum of the horseshoe crab (Limulus polyphemus) exoskeleton, eyes, arthrodial membrane, and base of the telson were documented in a population of wild caught laboratory animals. The disease can lead to loss of tissue structure and function, deformed shells, abnormal molting, loss of ocular structures, erosion of interskeletal membranes, and cardiac hemorrhage. Microscopy, histopathology, and in vitro culture confirmed the causative agent to be a green algae of the family Ulvaceae. Further research may explain how green algae overcome horseshoe crab innate immunity leading to external and internal damage.This study was supported in part by the Frederick B. Bang Fellowship and a grant from the Division of Research Resources, National Institutes of Health (P-40-RR1333-07)

Interrelationship between contractility, protein synthesis and metabolism in mantle of juvenile cuttlefish (Sepia officinalis)

Young juvenile cuttlefish (Sepia officinalis) can grow at rates as high as 12% body weight per day. How the metabolic demands of such a massive growth rate impacts muscle performance that competes for ATP is unknown. Here, we integrate aspects of contractility, protein synthesis, and energy metabolism in mantle of specimens weighing 1.1 g to lend insight into the processes. Isolated mantle muscle preparations were electrically stimulated and isometric force development monitored. Preparations were forced to contract at 3 Hz for 30 s to simulate a jetting event. We then measured oxygen consumption, glucose uptake and protein synthesis in the hour following the stimulation. Protein synthesis was inhibited with cycloheximide and glycolysis was inhibited with iodoacetic acid in a subset of samples. Inhibition of protein synthesis impaired contractility and decreased oxygen consumption. An intact protein synthesis is required to maintain contractility possibly due to rapidly turning over proteins. At least, 41% of whole animal ṀO2 is used to support protein synthesis in mantle, while the cost of protein synthesis (50 μmol O2 mg protein-1) in mantle was in the range reported for other aquatic ectotherms. A single jetting challenge stimulated protein synthesis by approximately 25% (2.51-3.12% day-1) over a 1 h post contractile period, a similar response to that which occurs in mammalian skeletal muscle. Aerobic metabolism was not supported by extracellular glucose leading to the contention that at this life stage either glycogen or amino acids are catabolized. Regardless, an intact glycolysis is required to support contractile performance and protein synthesis in resting muscle. It is proposed that glycolysis is needed to maintain intracellular ionic gradients. Intracellular glucose at approximately 3 mmol L-1 was higher than the 1 mmol L-1 glucose in the bathing medium suggesting an active glucose transport mechanism. Octopine did not accumulate during a single physiologically relevant jetting challenge; however, octopine accumulation increased following a stress that is sufficient to lower Arg-P and increase free arginine.Agência financiadora Natural Sciences and Engineering Research Council of Canada (NSERC) NSERC CPB section of the Canadian Society of Zoologists Fundacao para a Ciencia e a Tecnologia (FCT) IF/00576/2014 Portuguese national funds from Programa Operacional Mar2020 (Portugal2020/FEAMP) - Project SEPIACUL 16-02-01-FMP-53 FCT through Plurennial UID/Multi/04326/2019 EMBRC. PT ALG-01-0145-FEDER-022121 Portuguese node of EMBRC-ERICinfo:eu-repo/semantics/publishedVersio

Blood gases, biochemistry and haematology of Galápagos hawksbill turtles (Eretmochelys imbricata)

The hawksbill turtle, Eretmochelys imbricata, is a marine chelonian with a circum-global distribution, but the species is critically endangered and has nearly vanished from the eastern Pacific. Although reference blood parameter intervals have been published for many chelonian species and populations, including nesting Atlantic hawksbills, no such baseline biochemical and blood gas values have been reported for wild Pacific hawksbill turtles. Blood samples were drawn from eight hawksbill turtles captured in near shore foraging locations within the Galápagos archipelago over a period of four sequential years; three of these turtles were recaptured and sampled on multiple occasions. Of the eight sea turtles sampled, five were immature and of unknown sex, and the other three were females. A portable blood analyzer was used to obtain near immediate field results for a suite of blood gas and chemistry parameters. Values affected by temperature were corrected in two ways: (i) with standard formulas and (ii) with auto-corrections made by the portable analyzer. A bench top blood chemistry analyzer was used to measure a series of biochemistry parameters from plasma. Standard laboratory haematology techniques were employed for red and white blood cell counts and to determine haematocrit manually, which was compared to the haematocrit values generated by the portable analyzer. The values reported in this study provide reference data that may be useful in comparisons among populations and in detecting changes in health status among Galápagos sea turtles. The findings might also be helpful in future efforts to demonstrate associations between specific biochemical parameters and disease or environmental disasters

Blood Gases, Biochemistry, and Hematology of Galapagos Green Turtles (Chelonia Mydas)

The green turtle, Chelonia mydas, is an endangered marine chelonian with a circum-global distribution. Reference blood parameter intervals have been published for some chelonian species, but baseline hematology, biochemical, and blood gas values are lacking from the Galapagos sea turtles. Analyses were done on blood samples drawn from 28 green turtles captured in two foraging locations on San Cristóbal Island (14 from each site). Of these turtles, 20 were immature and of unknown sex; the other eight were males (five mature, three immature). A portable blood analyzer (iSTAT) was used to obtain near immediate field results for pH, lactate, pO2, pCO2, HCO3−, Hct, Hb, Na, K, iCa, and Glu. Parameter values affected by temperature were corrected in two ways: (1) with standard formulas; and (2) with auto-corrections made by the iSTAT. The two methods yielded clinically equivalent results. Standard laboratory hematology techniques were employed for the red and white blood cell counts and the hematocrit determination, which was also compared to the hematocrit values generated by the iSTAT. Of all blood analytes, only lactate concentrations were positively correlated with body size. All other values showed no significant difference between the two sample locations nor were they correlated with body size or internal temperature. For hematocrit count, the iSTAT blood analyzer yielded results indistinguishable from those obtained with high-speed centrifugation. The values reported in this study provide baseline data that may be useful in comparisons among populations and in detecting changes in health status among Galapagos sea turtles. The findings might also be helpful in future efforts to demonstrate associations between specific biochemical parameters and disease

Whole blood fatty acid concentrations in the San Cristóbal Galápagos tortoise (Chelonoidis chathamensis)

To continue releasing San Cristóbal Galápagos tortoises housed in managed-care facilities at the Giant Tortoise Breeding Center of Galápagos National Park (Galapaguera de Cerro Colorado) to the Otoy Ecological Farm, health assessments and physical examinations were conducted. As a part of these wellness examinations, blood was drawn from 11 tortoises to analyze fatty acid concentrations. Fatty acid levels can provide insight into the nutritional profiles, immune status, and reproductive health of vertebrates. To the co-author’s knowledge, there is no current information about fatty acids in this species. It was hypothesized that there would be inherent differences based on the different geographic ranges, diets, sex, and age of turtles. It was noted that the ω-6/ω-3 ratio was higher for the breeding center than for the ecological farm and that overall polyunsaturated fatty acids (PUFAs) did not have any significant differences. The ω-6/ω-3 findings can contribute to a global picture of these fatty acids across taxa, as reptiles are underrepresented in this area of research. Additional results are a resourceful starting point for future investigations into how fatty acids are affected in Galápagos tortoises

Standardized Direct Observation Assessment Tool: Using a Training Video.

BACKGROUND: We developed a DVD training tool to educate physicians evaluating emergency residents on accurate Standardized Direct Observation Assessment Tool (SDOT) application. OBJECTIVE: Our goal was to assess whether this training video improved attendings\u27 and senior residents\u27 SDOT use. METHODS: Participants voluntarily completed SDOT evaluations based on a scripted test video. A DVD with positive and negative scenarios of proper SDOT use was viewed. It included education on appropriate recording of 26 behaviors. The test scenario was viewed again and follow-up SDOTs submitted. Performances by attendings and residents on the pre- and post-test SDOTs were compared. RESULTS: Twenty-six attendings and 26 senior residents participated. Prior SDOT experience was noted for 8 attendings and 11 residents. For 20 anchors, participants recorded observed behaviors with statistically significant difference on one each of the pretest (no. 20; p = 0.034) and post-test (no. 14; p = 0.041) SDOTs. On global competency assessments, pretest medical knowledge (p = 0.016) differed significantly between groups. The training intervention changed one anchor (no. 5; p = 0.035) and one global assessment (systems-based practice; p = 0.031) more negatively for residents. Recording SDOTs with exact agreement occurred 48.73% for attendings pretest and 54.41% post-test; resident scores were 45.86% and 49.55%, respectively. DVD exposure slightly raised attending scores (p = 0.289) and significantly lowered resident scores (p = 0.046). CONCLUSIONS: Exposure to an independently developed SDOT training video tended to raise attending scores, though without significance, while at the same time lowered senior resident scores statistically significantly. Emergency attendings\u27 and senior residents\u27 SDOT scoring rarely differed with significance; about half of anchor behaviors were recorded with exact agreement. This suggests senior residents, with appropriate education, may participate in SDOT assessment

Methods of body temperature assessment in Conolophus subcristatus, Conolophus pallidus (Galápagos land iguanas), and Amblyrhynchus cristatus X C. subcristatus hybrid

Since cardiovascular, respiratory, and metabolic systems of reptiles are affected by temperature, accurate measurements are of great importance in both captive husbandry and research. Ectothermic animals generally have core body temperatures close to ambient temperature but can differ from the immediate environment if they are using sunlight to thermoregulate. Many zoological facilities and exotic pet caregivers have begun using infrared temperature guns to assess ambient temperatures of reptile enclosures but there are currently few studies assessing the efficacy of these devices for measuring the body temperatures of reptiles. Conolophus subcristatus, Conolophus pallidus (Galápagos land iguanas), and Amblyrhynchus cristatus X C. subcristatus hybrid are robust land iguanas endemic to the Galápagos archipelago. By comparing the infrared body temperature measurements of land iguanas against virtual simultaneous collection of cloacal temperatures obtained using a thermocouple thermometer, we sought to assess the efficacy of this non-invasive method. We found that internal body temperature can be predicted with a high level of accuracy from three external body temperature sites, providing a good non-invasive method that avoids the capture of animals

Blood gases, biochemistry and haematology of Galápagos marine iguanas ( Amblyrhynchus cristatus )

The marine iguana, Amblyrhynchus cristatus, is an iconic lizard endemic to the Galápagos Islands of Ecuador, but surprisingly little information exists on baseline health parameters for this species. We analysed blood samples drawn from 35 marine iguanas captured at three locations on San Cristóbal Island. A portable blood analyser (iSTAT) was used to obtain near-immediate field results for pH, lactate, partial pressure of O2, partial pressure of CO2, bicarbonate (HCO3−), percentage O2 saturation, haematocrit, haemoglobin, sodium, potassium, ionized calcium and glucose. Parameter values affected by temperature were auto-corrected by the iSTAT. Standard laboratory haematology techniques were employed for differential white blood cell counts and haematocrit determination; resulting values were also compared with the haematocrit values generated by the iSTAT. Body temperature, heart rate, respiratory rate and body measurements were also recorded. Body length was positively correlated with several blood chemistry values (HCO3− and glucose) and two haematology parameters (haemoglobin and manually determined haematocrit). A notable finding was the unusually high blood sodium level; the mean value of 178 mg/dl is among the highest known for any reptile. This value is likely to be a conservative estimate because some samples exceeded the maximal value the iSTAT can detect. For haematocrit determination, the iSTAT blood analyser yielded results significantly lower than those obtained with high-speed centrifugation. The values reported in this study provide baseline data that may be useful in comparisons among populations and in detecting changes in health status among marine iguanas affected by natural disturbances or anthropogenic threats. The findings might also be helpful in future efforts to demonstrate associations between specific biochemical parameters and disease