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Objective: The objective of this study was to analyze the use of rebound tonometry, as a noninvasive diagnostic tool, in arachnids., Methods: 5 juvenile (yearling) female and 1 juvenile male curlyhair tarantulas (Tliltocatl albopilosus, previously Brachypelma albopilosum) were used to track estimated ventral opisthosoma pressures over a 9-month period of time. Younger, growing animals were selected as they are more likely than adults to go through multiple molts throughout the 9 months of the study length. An iCare TONOVET TV01 rebound tonometer was used to measure the estimated ventral opisthosoma pressures of the spiders. Measurements were obtained from the ventral opisthosoma, 1 of the thinnest areas of exocuticle throughout the body. Readings were obtained once per week from the ventral opisthosoma for the first 2 months, then once every 2 weeks for 1 month, then back to once per week for the remainder of the study. Additional measurements were obtained following each ecdysis, after a 2% body weight sampling of hemolymph, and at the end of the study to evaluate readings in response to induction of general anesthesia with 5% isoflurane gas and oxygen flow at 2 L/min for 10 minutes., Results: The average of all estimated ventral opisthosoma pressure when spiders were not in molt was 26.19 mm Hg (SD, 3.54), with a statistically significant decrease postmolt to an average of 15.31 mm Hg (SD, 3.81), followed by a gradual increase back to premolt pressures over a 3-week period with an average of 22 days (SD, 1.93). Estimated ventral opisthosoma pressures decreased post hemolymph removal. There is not sufficient evidence that estimated ventral opisthosoma pressures changed over time following the anesthesia., Clinical Relevance: This study demonstrates that tonometry can be used to generally assess the estimated ventral opisthosoma pressure, which could correlate with where a spider is in an ecdysis cycle.

Objective: To evaluate the pathologic ocular changes in a population of aquarium-housed Monodactylus argenteus and investigate potential underlying causes for the large number of affected fish in this exhibit., Animals: 11 Monodactylus argenteus were evaluated from a shoaling ring exhibit within an aquarium, and 19 control fish without ocular abnormalities were obtained from commercial fish suppliers., Methods: Physical and ocular examinations were performed antemortem. Postmortem samples of liver, heart, dorsal epaxial muscle, and lenses of affected and control fish were analyzed for amino acid profiles. The aqueous humor from affected and control fish was collected postmortem, and osmolality was analyzed. Tissues from affected and control fish were submitted for histopathology., Results: Ocular abnormalities in affected fish included corneal lesions, cataracts, lens capsule rupture, and unilateral left-sided lens luxation and buphthalmos. Lens luxation and buphthalmos were directly correlated. Aqueous humor osmolality in control fish differed significantly compared to affected fish but was not correlated to lens abnormality score. Affected fish had significantly lower lens concentrations of arginine, asparagine, glycine, isoleucine, serine, and tyrosine than control fish. One affected fish had severe buphthalmos, cataracts, and panophthalmitis caused by gram-positive cocci., Conclusions: Cataracts and traumatic lesions were common in affected M argenteus. Dietary amino acid content and aqueous humor osmolality imbalances were not likely a cause of the cataracts. Differences in lens amino acid concentrations between affected and control fish may be related to cataract formation., Clinical Relevance: Consideration of species-appropriate tank parameters may mitigate ocular lesions including cataracts in aquarium fish.

Invasive alien species control is recognized worldwide as a priority action to preserve global biodiversity. However, a lack of general life history knowledge for threatened species can impede the effectiveness of conservation actions. Galápagos pink land iguanas ( Conolophus marthae ) are endemic to Wolf Volcano, Galápagos, Ecuador. These iguanas are threatened by invasive alien species, particularly feral cats, that may affect their small population size. To guarantee the long-term survival of C. marthae , the Galápagos National Park Directorate is considering, along with an ongoing campaign of feral cat control, the implementation of a head-start program. However, the success of this management strategy necessarily relies on the identification of pink iguana nesting grounds, which were still unknown at the onset of this study. We modeled the movement patterns of male and female iguanas during the reproductive season, using location data collected from custom-made remote tracking devices installed on adult pink iguanas in April 2021. We first calculated for each individual the vector of distances from its starting location, which was defined as net displacement. We then used net displacement as the response variable in a generalized additive mixed model with day of the year as the predictor. Based on the hypothesis that males and females may behaviorally differ after mating, we looked for female-specific migratory behavior suggesting females were moving toward nesting areas. The results obtained confirmed our hypothesis, as females exhibited a distinct migratory behavior, reaching a small plateau area inside of Wolf Volcano's caldera and ca. 400 m below the volcano's northern rim. Moreover, once inside the caldera, females displayed a more aggregated distribution pattern. The movement data obtained allowed Galápagos National Park rangers to locate individual pink iguana nests and subsequently to sight and collect the first observed hatchlings of the species. This work constitutes a necessary baseline to perform dedicated studies of pink iguana nests and emerging hatchling iguanas, which is an essential step toward the development of an effective head-start program.

As the analysis of blood metabolites has become more readily accessible thanks to the use of point-of-care analyzers, it is now possible to evaluate stress level of wild animals directly in the field. Lactate is receiving much attention as a good stress level proxy in individuals subjected to capture, manual restraint, and data sampling in the wild, and appropriate protocols to maintain lactate values low should be preferred. In this study we compare how two different capture methodologies, hand grab vs. noose pole, affect the variation of blood lactate values in Cyclura carinata iguanas when captured for sampling. We used blood lactate concentration, measured immediately upon- and 15 min after-capture, as a proxy for stress level. While the primary goal of this work is to determine the least stressful capture methodology to be favored when sampling this and other wild iguanas, we also evaluated additional baseline physiological parameters relevant to the health and disease monitoring for this species. Our results show that while initial lactate values level-out in sampled individuals after 15 min in captivity, regardless of the capture methodology, rock iguanas captured by noose pole showed significantly higher lactate concentration and increased heartbeat rate immediately after capture. While the overall health evaluation determined that all analyzed individuals were in good health, based on our results we recommend that, when possible, hand capture should be preferred over noose pole when sampling wild individuals., Competing Interests: The authors declare that they have no competing interests., (© 2024 Colosimo et al.)

Virus surveillance in wildlife is important to understanding ecosystem health, taxonomy, and evolution. Nevertheless, viruses in reptiles, and specifically in squamates, continue to be understudied. Herein, we conducted a health assessment on the critically endangered Galapagos pink land iguana (Conolophus marthae) and the vulnerable Galapagos land iguana (Conolophus subcristatus). We collected oral and/or cloacal swabs from 47 clinically healthy iguanas and tested for adenovirus (cloacal swabs, n = 47) and herpesvirus (oral swabs, n = 45) using broad-spectrum PCRs. Two out of 38 (5.3 %) Galapagos pink land iguanas tested positive for herpesvirus, while no herpesvirus was detected in all Galapagos land iguanas (n = 7). Both herpesviral sequences were identical between them and divergent (61.9 % amino acid identity) when compared to the closest herpesvirus sequences available in GenBank/EMBL/DDBJ. The genetic distance between this and other herpesviruses is consistent with its classification as a novel virus species. All iguanas were negative for adenovirus. This is the first description of a herpesvirus in iguanas of the Galapagos islands, and the first report of a potential pathogen for the iconic Galapagos pink land iguana. Further research is needed to understand the implications of this virus in the conservation and management of one of the most endangered iguana species in the world., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)

Insects are increasingly common in households, zoological collections, research, and food industries. Increased knowledge of insect health parameters is necessary to ensure proper welfare. The study goal was to develop a hemolymph collection technique and report hemolymph serum biochemical parameters for the thorny devil stick insect ( Eurycantha calcarata ). Clinically healthy adult stick insects (5 males and 14 females, 15.1-24.7 g) were enrolled. Four collection techniques were evaluated. Hemolymph collection was unsuccessful in males, but was successful in females by using a single technique. The insect was manually restrained in an elevated position and an 18-ga × 2.54-cm needle was used to puncture the membrane just caudal to the third pair of legs. With the puncture site directed ventrally, ≤1 ml of hemolymph was collected via gravity and allowed to visibly clot. The sample was then centrifuged, and the serum was separated. Serum samples were individually analyzed (Avian/Reptilian Profile Plus, VetScan VS2, Abaxis, Inc, Union City, CA 94587, USA). Fourteen samples (0.2-1.0 ml) were collected from 14 females. Median (minimum-maximum) parameters included the following: aspartate aminotransferase 12 (0-45) U/L, creatinine kinase 25 (0-76) U/L, uric acid 7.5 (3.1-13.7) mg/dl, glucose 12 (8-22) mg/dl, calcium 18.6 (17.2-19.4) mg/dl, phosphorus 15.0 ( n = 1) or >30.0 ( n = 13) mg/dL, total protein 2.7 (1.6-2.9) g/dL, albumin 0.9 (0.2-1.2) g/dL, globulin 1.7 (1.6-1.8) g/dL ( n = 6) or not quantified ( n = 8), potassium 10.6 (9.0-11.8) mmol/L, sodium < 100 mmol/L, and bile acids 0 lmol/L. This is the first report of biochemistry parameters in clinically healthy female stick insects. Larger sample sizes are needed to establish statistically valid reference ranges. Hemolymph collection techniques for male stick insects warrant further investigation.

Background: Dried blood spot (DBS) technology is valuable in providing simple means of storing blood samples from wildlife with small blood volumes. Methods designed for heavy metal analysis on DBS become more useful if extended to elements of nutritional significance., Purpose: (1) Development of procedures for measuring Mn, Fe, Co, Cu, Zn, Se and Mo in DBS; (2) use the designed methods in health assessments of Galápagos land iguanas (Conolophus species)., Procedures: Elements were measured by inductively coupled plasma/mass spectrometry (ICP-MS) following acid digestion of whole blood or DBS from the same animal for direct comparison. Study animals comprised free-ranging iguanas from separate islands in the Galápagos archipelago., Main Findings: DBS spikes (Mn, Fe, Co, Cu, Zn, Se and Mo) demonstrated accuracy to ∼100 ppb; reporting limits were set there except for Fe and Zn which were set at 1000 ppb. Plasma samples - generally preferable for nutritional element diagnostics - were submitted from Galápagos land iguanas along with DBS as part of a large-scale health assessment. In plasma versus DBS concentration comparisons, Fe, Cu, Se and Mn correlated well with R^2 values of 0.799, 0.818, 0.896 and 0.899, respectively, and slopes ranging 0.88 - 1.3. Co and Zn showed greater scatter. Mo had insufficient points above its reporting limit and offered advantages for toxicity assessments. Bland-Altman diagrams showed flat scatter between 2x standard deviation boundaries with no undue trends except for Mn which had few points above its reporting limit. Bias, defined as the average difference [DBS - plasma] divided by the average value, was relatively low throughout, with values of - 19.3 % (Fe), - 48.7 % (Co), - 19.6 % (Cu), - 6.9 % (Zn), - 21.4 % (Se) and + 40.7 % (Mn). Normal distribution assessment of iguana Cu, Zn, Se and Fe plasma values showed unanticipated divergences between two species., Conclusions: The DBS approach for nutritional element analysis offers a suitable methodology for determining crucial elements Mn, Fe, Co, Cu, Zn, Se, and Mo in veterinary samples. Analyses of samples from Conolophus species revealed interesting divergences particularly for Cu, Zn, Se and Fe, elements generally associated with defense against oxidative stress., Competing Interests: Declaration of Competing Interest The authors confirm that there are no existing conflicts of interest related to the presented work., (Copyright © 2023 Elsevier GmbH. All rights reserved.)

Forty-seven free-ranging sea turtles (46- Chelonia mydas, 1- Eretmochelys imbricata) were examined via novel use of an endoscopy combined with a rectal enema to obtain large fecal sample volumes. The cloaca was insufflated using an endoscope, after which the bladder and rectum separated, allowing access to the colon. Environmental conditions and location influenced the performance of the procedure initially, but after several attempts the procedure was successfully initiated. In all cases, fecal samples were obtained, and the animals were released to their respective locations. Fecal sample collection using this approach enhances the ability to obtain diagnostic information and perform other scientific analyses of sea turtles., (© 2023 The Authors. Zoo Biology published by Wiley Periodicals LLC.)

Background: Antimicrobial resistance (AR) has led to increasing human and animal morbidity and mortality and negative consequences for the environment. AR among Escherichia coli (EC) is on the rise, with serious concerns about extended-spectrum β-lactamase-producing E. coli (ESBL-EC). In the Galápagos Islands, where antimicrobials are available without a prescription, growing demands for food production can drive antimicrobial use. Food producing animals are at the interface of wildlife and environmental health on the smallest human-inhabited Galápagos Island, Floreana. We sought to determine if ESBL-EC were present in Floreana Island farm animal species and nearby wildlife and the relatedness of ESBL-EC isolates identified. Materials and Methods: During July 4-5, 2022, we visited 8 multispecies farms, representing 75% of food-producing animal production on Floreana, and collected 227 fecal samples from farm animals and wildlife. Each sample was plated on MacConkey agar supplemented with cefotaxime (4 μg/mL). Results: ESBL-EC was isolated from 20 (9%) fecal samples collected from pigs ( N  = 10), chickens ( N  = 6), wildlife ( N  = 3), and dog ( N  = 1). All ESBL-EC isolates were from samples taken at three (38%) of the eight farms. Fifteen (75%) of the ESBL-EC isolates were from a single farm. All ESBL-EC isolates were multidrug resistant. The most prevalent ESBL genes belonged to the bla CTX-M group. Among the typeable isolates from the farm with the largest proportion of ESBL-EC isolates ( N  = 14), we observed nine unique pulsed-field gel electrophoresis (PFGE) patterns, with identical patterns present across pig and chicken isolates. PFGE patterns in the three farms with ESBL-EC isolates were different. Conclusions: These results lend support for future routine AR monitoring activities at the livestock-wildlife interface in Galápagos to characterize potential interspecies transmission of AR bacteria and AR genes in this unique protected ecosystem, and the related human, animal, and environmental health impacts, and to formulate interventions to reduce AR spread in this setting.

Objective: To compare dexmedetomidine-ketamine (DK; 0.1 and 10 mg/kg, respectively) with midazolam (M; 1.0 mg/kg) or 0.9% sodium chloride (S; 0.2 mL/kg) administered IM in the forelimb (F) or hindlimb (H) in eastern box turtles (Terrapene carolina carolina)., Animals: 20 clinically healthy, captive adult eastern box turtles., Methods: In a randomized, blinded, complete crossover study with 1-week washout periods, turtles were administered each of 3 treatments: F-DKS, F-DKM, or H-DKM. Palpebral reflex, muscle tone, and withdrawal responses were serially assessed and used to calculate cumulative sedation scores at each 5-minute time point. The ability to intubate was evaluated. At 60 minutes, atipamezole (1.0 mg/kg) and either flumazenil (F-DKM, H-DKM; 0.05 mg/kg) or 0.9% sodium chloride (F-DKS; 0.5 mL/kg) were administered IM., Results: All treatments resulted in clinically relevant anesthetic effects. F-DKM produced significantly higher sedation scores than H-DKM or F-DKS at all time points between 10 and 60 minutes (P < .05). Sedation score variability was observed with all treatments with significantly higher variability for H-DKM (P < .05). Intubation was successful in 32, 89, and 11% of turtles in F-DKS, F-DKM, and H-DKM, respectively. Median (range) recovery time was 10 (5-22), 16 (7-45), and 12 (4-28) minutes for F-DKS, F-DKM, and H-DKM, respectively., Clinical Relevance: In eastern box turtles, forelimb dexmedetomidine-ketamine resulted in clinically relevant anesthetic effects that were heightened with the addition of midazolam. Hindlimb administration of midazolam-dexmedetomidine-ketamine resulted in reduced and more variable anesthetic effects compared to forelimb administration, supporting a hepatic first-pass effect.

Invertebrates are a diverse group of animals that make up the majority of the animal kingdom and encompass a wide array of species with varying adaptations and characteristics. Invertebrates are found in nearly all of the world's habitats, including aquatic, marine, and terrestrial environments. There are many misconceptions about invertebrate sentience, welfare requirements, the need for environmental enrichment, and overall care and husbandry for this amazing group of animals. This review addresses these topics and more for a select group of invertebrates with biomedical, economical, display, and human companionship importance.

Invertebrates, including arachnids, are a common taxon in zoological collections. Invertebrate medicine and pathology are emerging subspecialties, but there is limited reference material or published resources describing histologic lesions in arachnids. Histopathology of 26 captive arachnids (20 spiders and 6 scorpions) from institutional collections was reviewed. Most animals were found dead with limited clinical signs. Tissues evaluated included body wall (cuticle and epidermis), skeletal muscle, book lungs, digestive tract (pharynx, esophagus, sucking stomach, midgut tube, midgut diverticula, and stercoral pocket), central and peripheral nervous system, heart, hemolymph vessels and sinuses, Malpighian tubules, coxal glands, and gonads. Inflammation was frequent (24/26, 92%), and seen in multiple organs (18/24, 75%) with the midgut diverticulum most commonly affected (14/24, 58%) followed by the book lungs (13/24 arachnids, 54%), and body wall (8/24 arachnids, 33%). Inflammation comprised hemocyte accumulation, hemocytic coagula, melanization, and nodulation. Infectious agents, including bacteria (11/26, 42%), fungi (10/26, 38%), and parasites (2/26, 8%), were seen within inflammatory aggregates. Coinfection with multiple infectious agents was common (6/24, 25%). No etiologic agent was identified in 7/24 (29%) cases with inflammatory lesions. Lesions suggestive of decreased nutritional status or increased metabolic rate included midgut diverticula atrophy in 11/26 (42%) animals and skeletal muscle atrophy in 6/26 (23%) animals. Atrophic lesions were seen in combination with infection (8/11, 73%), pregnancy (2/11, 18%), male sex (2/11, 18%), or without other lesions (1/11, 9%). Other suspected contributors to death included dysecdysis-associated trauma (2/26, 8%) and uterine intussusception (1/26, 4%). No animals had neoplasia.

The red-billed tropicbird, Phaethon aethereus , is a species of seabird native to the Galápagos archipelago, and widely distributed across the neotropics. General health, blood chemistry, and haematology parameters have not been published for this species. Blood analyses were performed on samples drawn from 51 clinically healthy red-billed tropicbirds captured from their burrows at Islote Pitt on San Cristóbal Island in July, 2016 (21) and Daphne Major Island in June, 2017 (30). In the field, a point of care blood analyser (iSTAT) was used to obtain results for HCO 3- , pH, pCO 2 , pO 2 , TCO 2 , iCa, Na, K, Cl, Hb, HCT, anion gap, creatinine, glucose and urea nitrogen. Additionally, a portable Lactate Plus TM analyser was used to measure blood lactate, and blood smears were also created in situ . The blood slides were used to estimate leukocyte counts and 100-cell differentials. Alongside these biochemistry and haematology parameters, average heart rate, respiratory rate, body temperature and scaled mass index (calculated from weight and a body measurement) were compared to determine the standard measurements for a healthy individual. The baseline data, and reference intervals reported in this paper are essential to detecting changes in the health of red-billed tropicbirds in the future., Competing Interests: The authors declare there are no competing interests., (©2023 Skehel et al.)

Objective: To describe the clinical features, treatment, and outcomes of wild freshwater turtles with fishing hook injuries., Animals: 126 wild turtles residing in central North Carolina that were presented to a wildlife rescue clinic., Methods: Medical records from July 1997 to July 2022 were reviewed, and data were collected and analyzed., Results: The most common species presenting for a fishhook injury was the yellow-bellied slider (Trachemys scripta scripta) (n = 69/126 [54.8%]; 95% CI, 45.7 to 63.6). The most common location identified was the oral cavity (n = 77/140 [55%]; 95% CI, 46.4 to 63.4) and the most common removal method was retrograde removal after cutting the barb off of the hook (76/120 [63.3%]; 95% CI, 54.1 to 71.9). Fishhooks embedded in the esophagus had a significantly higher chance of complications affecting recovery (OR estimate, 3.49; 95% CI, 1.07 to 11.38). There was no significant increase in mortality associated with the location of the injury; however, there was a significant increase in mortality in patients that experienced complications (P < 0.001). The time in care ranged from 1 to 150 days (median, 16 days). Of the turtles evaluated, 10.8% (n = 12/111; 95% CI, 5.7 to 18.1) were euthanized or died after treatment and 89.2% (99/111; 95% CI, 81.9 to 94.3) were released., Clinical Relevance: These findings describe various successful techniques to remove fishhooks from turtles. While no superior treatment was identified, considerations should be taken to provide patient comfort, decrease injury-associated complications, and shorten recovery time by using minimally invasive techniques. Overall, freshwater turtles with fishhook injuries have a high release rate even when the injuries are severe.

Giant Pacific octopuses ( Enteroctopus dofleini ) (GPOs) are commonly housed in zoos or aquaria, and sedation, anesthesia, and/or euthanasia may be indicated for a variety of reasons. Despite this need, evidence-based data on best practices is limited and focuses on smaller or more tropical species. The objectives of this study were to survey the aquatic community regarding the husbandry and veterinary care of GPOs, with a specific focus on anesthetic and euthanasia protocols. A two-part web-based survey was distributed to four aquatic and/or veterinary email listservs. Individuals from fifty-two institutions participated in phase one. Results documented that 40 (78 percent) participating institutions currently house GPOs, with most housing one and nine institutions housing two to three GPOs. The median (range) habitat volume is 5405 (1893-16,465) L, and 78 percent of systems are closed. Of the institutions surveyed, 23 have anesthetized or sedated a GPO for nonterminal procedures, including wound care, biopsies, and hemolymph collection. Reported methods of sedation or anesthesia include magnesium chloride, ethanol, isoflurane, tricaine methanesulfonate (MS-222), magnesium sulfate, benzocaine, and dexmedetomidine. Drugs or methods used for euthanasia include magnesium chloride, ethanol, mechanical decerebration, pentobarbital, isoflurane, MS-222, magnesium sulfate, benzocaine, potassium chloride, dexmedetomidine, and freezing. Reported observed side effects include ineffectiveness or inadequate sedation, inking, prolonged drug effects, and behavior changes. Survey data have the potential to guide the husbandry and veterinary care of GPOs and build the framework for future prospective studies on GPO sedation and anesthesia.

Toxoplasma gondii is a zoonotic intracellular parasite of particular concern in the conservation of wildlife due to its ability to infect all homeotherms and potentially cause acute fatal disease in naive species. In the Galapagos (Ecuador), an archipelago composed of more than a hundred islets and islands, the presence of T. gondii can be attributed to human-introduced domestic cats, but little is known about its transmission in wildlife populations. We compared the prevalence of antibodies against T. gondii in sympatric Galapagos wild bird species that differ in diet and contact with oocyst-contaminated soil to determine the relative importance of trophic habits as an exposure factor. Plasma samples were obtained from 163 land birds inhabiting Santa Cruz, one of the cat-inhabited islands, and from 187 seabirds breeding in cat-free surrounding islands (Daphne Major, North Seymour, and South Plaza). These samples were tested for the presence of T. gondii antibodies using the modified agglutination test (MAT ≥ 1:10). All seven species of land birds and 4/6 species of seabirds presented seropositive results. All great frigatebirds (Fregata minor) (N = 25) and swallow-tailed gulls (Creagrus furcatus) (N = 23) were seronegative. Prevalence ranged from 13% in Nazca boobies (Sula granti) to 100% in Galapagos mockingbirds (Mimus parvulus). It decreased from occasional carnivores (63.43%) to granivores-insectivores (26.22%), and strict piscivores (14.62%). These results indicate that the consumption of tissue cysts poses the highest risk of exposure to T. gondii for Galapagos birds, followed by the ingestion of plants and insects contaminated by oocysts as important transmission pathways., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Mosquera et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

The Galapagos sea lion (Zalophus wollebaeki), an endemic and endangered pinniped, faces an increasing threat due to infectious diseases related to domestic animals. Dirofilaria immitis, the parasite responsible for canine heartworm disease, is one such threat, as canine infections on the archipelago have been documented. We used a canine heartworm antigen test kit to analyze the blood from 25 juvenile Galapagos sea lions for D. immitis. Two (8%) sea lions tested positive for D. immitis antigen. Using morphologic and genetic assessments, we evaluated 20 filarial-like worms collected from within the heart of an adult male Galapagos sea lion during a previous routine postmortem examination. The intracardiac worms were morphologically consistent with adult D. immitis, and sequence analysis of targeted PCR amplicons confirmed their identity. This is the first report of D. immitis infection in Galapagos sea lions, which could become a major health problem for these pinnipeds. Further studies are necessary to confirm the level of threat from this parasite; however, widespread adoption of routine heartworm testing, prevention, and treatment in the canine population, and the control of mosquitos, could potentially reduce the disease impact on this endangered pinniped species., (© Wildlife Disease Association 2023.)

Free coelomic fluid can be a physiologic or pathologic finding in snakes. In this study, ultrasonography was used to assess the presence, volume, and type of coelomic fluid in 18 (16 females, 2 males) clinically healthy corn snakes ( Pantherophis guttatus ) using a semiquantitative scoring system. Each snake was divided into five equal regions (R1-R5) by length (from rostrum to vent) and fluid volume was scored on a scale of 0 to 4. Coelomic fluid was sampled when possible and cytologic analysis was performed. The majority of snakes (16/18) were found to have some degree of free coelomic fluid. The coelomic fluid samples that were collected ( n = 6) were classified as transudates, acellular or primarily lymphocytic. Fluid was most likely to be found in R3 compared with all other regions and least likely to be found in R1 compared with R2, R3, and R4. A higher score of volume was found in R3 compared with R1 and R5. This study describes the distribution and abundance of coelomic fluid in snakes, as well as a method for point of care ultrasound (POCUS) in this species.

Lateral line depigmentation (LLD) is a common condition in managed tropical saltwater fish, and treatment is somewhat elusive. Naltrexone, an opioid receptor antagonist, enhances epithelial cell replication, cytokine production, and angiogenesis to stimulate wound healing in mice. A treatment trial with 11 palette surgeonfish with LLD was performed. Seven fish underwent a single topical treatment of a mixture of 4 mg naltrexone and 10 g iLEX petroleum paste applied topically to LLD lesions. Four additional fish served as controls: two received only topical iLEX and two received no treatment. Severity of disease was scored on a 0-3 scale. Inflammatory response was gauged on a separate 0-3 scale for 5 d after treatment based on severity of erythema, as seen in a clinical case performed prior to this study. After 11 days, four affected animals that lacked an inflammatory response after naltrexone topical treatment were administered a single dose of intralesional 0.04% naltrexone (4 mg diluted into 10 ml saline). Lesions on all fish were photographed and measured at day 33. Clinical improvements in lesion size and pigmentation were apparent following topical naltrexone therapy in fish with severe lesions. Although these cases are promising, more data are needed to further evaluate the effectiveness of naltrexone 0.04% in treating LLD lesions in palette surgeonfish.

Globally to date, established international standards for animal welfare, a priority of sustainable agriculture, have primarily focused on large-scale producers. However, across Latin America, including in Ecuador's Galápagos Islands, smallholder farms play a critical role in food safety and security. We assessed five basic animal welfare measures (feed and water access, shelter availability and housing systems, animal health management, animal behavior, and timely euthanasia) for poultry, pigs, and cattle on Floreana Island, Galápagos. Utilizing assessment standards from multiple US sources and international standards, we developed a questionnaire and used it to conduct in-depth interviews during 4-5 July 2022 with eight participating producers, representing 75% of animal agriculture on Floreana. While we identified opportunities to enhance competencies in animal health management and timely euthanasia, farms performed well in the other assessed measures. Future work should promote knowledge transfer and in-country capacity building in farm biosecurity, access to veterinary care, antimicrobial resistance surveillance, and euthanasia methods. Efforts to positively impact smallholder farm livelihoods in Galápagos-one of the most biodiverse and protected ecosystems on the planet-will sustainably support human health through the interconnected realms of animal health and welfare, wildlife and environmental health, and food safety and security.

Objective: To evaluate latency and duration of a brachial plexus block technique in eastern box turtles performed with 2% lidocaine at three dose rates., Study Design: Prospective, randomized, blinded crossover study., Animals: Adult eastern box turtles, two for drug dose evaluation and a group of six (three male, three female) weighing 432 ± 40 g (mean ± standard deviation) for the main study., Methods: Animals were randomly assigned to four brachial plexus blocks with lidocaine at 5, 10 and 20 mg kg -1 or 0.9% saline (treatments LID5, LID10, LID20 and CON, respectively), separated by 1 week. Treatment side was randomized and blocks were performed unilaterally. Baseline observations of mentation, heart rate (HR), respiratory rate (f R ), skin temperature and limb response to manipulation or toe pinch were evaluated. Assessments were made every 10 minutes until 1 hour of normal sensory and motor function to the treated thoracic limb, or for a total of 2 hours if no block was evident., Results: Motor and sensory blockade was achieved in treatments LID10 and LID20 in one turtle, with a latency of 10 minutes and duration of 50 minutes for both doses. Raising of the ipsilateral lower palpebra occurred with both blocks. Turtles administered lidocaine experienced higher HR compared with CON, and HR decreased over time for all individuals. Mentation and f R were not changed with any lidocaine dose., Conclusions: The technique was unreliable in producing brachial plexus motor and sensory blockade at the lidocaine doses evaluated in this study. HR was higher in lidocaine-administered turtles but remained within normal limits for the species. No change in mentation or f R was observed among treatments., Clinical Relevance: General anesthesia with systemic analgesia is recommended for surgical procedures involving the chelonian thoracic limb. Further studies are needed to optimize a brachial plexus block in this species., (Copyright © 2022 Association of Veterinary Anaesthetists and American College of Veterinary Anesthesia and Analgesia. Published by Elsevier Ltd. All rights reserved.)

Welfare considerations and regulations for invertebrates have lagged behind those for vertebrates, despite invertebrates comprising more than 95% of earth's species. Humans interact with and use aquatic invertebrates for exhibition in zoos and aquaria, as pets, research subjects, and important food sources. Recent research has indicated that aquatic invertebrates, in particular cephalopod mollusks and decapod crustaceans, experience stress and may be able to feel pain. With this article, we present results of a survey on attitudes of aquatic animal health professionals toward aquatic invertebrate welfare and provide practical recommendations for advancing aquatic invertebrate welfare across four areas of opportunity: use of anesthesia, analgesia, and euthanasia; development of less invasive diagnostic and research sampling methods based on 3R principles; use of humane slaughter methods for aquatic invertebrates; and reducing impacts of invasive procedures in aquaculture and fisheries. We encourage consideration of these opportunities to achieve far-reaching improvements in aquatic invertebrate welfare., Competing Interests: Author AN is employed by ZooQuatic Laboratory, LLC. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Wahltinez, Stacy, Hadfield, Harms, Lewbart, Newton and Nunamaker.)

Pentastomes are crustacean endoparasites in the lungs of herpetofauna and rarely mammals and birds. To date, the only pentastomes reported in chelonians, all aquatic turtles, are Diesingia megastomum from Brazil and Pelonia africana from South Africa. In March 2021, 185 juvenile tortoises (Chelonoidis niger) were confiscated after an attempted illegal exportation from the Galápagos. It is believed the tortoises were removed from nests on Santa Cruz Island. The young tortoises were individually wrapped in plastic and at seizure ten were dead and another 25 tortoises subsequently died. One of the tortoises, estimated to be 3 months old, had 11 pentastomes in the lungs. The pentastomes were identified as a Raillietiella sp. based on morphology. The specimens had a claviform body that tapers into a bifid tail, a 165.3 μm × 92.1 μm buccal cadre, and sharp tipped anterior and posterior hooks with the posterior hooks being larger than the anterior. The males have smooth copulatory spicules with a rounded, smooth base. All females were gravid. Molecular analysis confirmed that the parasites were a Raillietiella sp. Based on the COI gene, it was most similar (82.7% identical) to an undescribed Raillietiella species from a Caribbean anole (Anolis cristatellus) found in Florida followed by R. hebitihamata (81%), R. indica (80.7%), and R. orientalis (78.8-80.7%). Based on the 18S rRNA gene sequence (1799 bp), it was most similar (99.3% identical) to two undescribed Raillietiella species followed by R. aegypti from a berber skink (Eumeces schneideri) from Saudia Arabia. Phylogenetically, with both molecular targets, the Raillietiella sp. from the Galápagos tortoise grouped with other Raillietiella spp. and was basal within the group. Currently, the origin of this parasite (native to Galápagos or introduced) and the life cycle are unknown. Because some pentastome species, especially when in aberrant hosts, can be pathogenic, additional studies of parasites in native and introduced reptile and amphibian species in the Galápagos are needed to better understand the risk this parasite poses to Galápagos tortoises., Competing Interests: Declaration of Competing Interest We wish to confirm that there are no known conflicts of interest associated with this publication and there has been no significant financial support for this work that could have influenced its outcome. We confirm that the manuscript has been read and approved by all named authors and that there are no other persons who satisfied the criteria for authorship but are not listed. We further confirm that the order of authors listed in the manuscript has been approved by all of us. We confirm that we have given due consideration to the protection of intellectual property associated with this work and that there are no impediments to publication, including the timing of publication, with respect to intellectual property. In so doing we confirm that we have followed the regulations of our institutions concerning intellectual property. We understand that the Corresponding Author is the sole contact for the Editorial process (including Editorial Manager and direct communications with the office). He/she is responsible for communicating with the other authors about progress, submissions of revisions and final approval of proofs. We confirm that we have provided a current, correct email address which is accessible by the Corresponding Author and which has been configured to accept email from myabsley@uga.edu, (Copyright © 2022 Elsevier B.V. All rights reserved.)

The Galapagos sea lion ( Zalophus wollebaeki ) is an endemic pinniped to the Galapagos archipelago, and like most wild mammals, is at risk for anemia due to trauma, infectious disease, and poor nutrition. This study evaluated the health status of 26 juvenile Galapagos sea lions on the island of San Cristobal prior to evaluating 100 crossmatch combinations. On evaluation, all but one sea lion had no major systemic abnormalities. Of the 100 crossmatches performed, 23% had minor reactions. The most significant reaction was weak macroscopic agglutination found in 4% of samples. The small percentage of agglutination reactions suggests a small proportion of naturally occurring alloantibodies in this species and may be consistent with a low risk of acute immune-mediated hemolytic transfusion reaction., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Gregory, Parker, Deresienski, Alarcón-Ruales, Muñoz-Pérez, Torres, Gavilanes, Lewbart and Páez-Rosas.)

The thorny devil stick insect ( Eurycantha calcarata ) is a common invertebrate maintained under human care. Blood (hemolymph) transfusions are a widely used therapeutic tool in other species, but investigation in terrestrial arthropods remains scarce. Study objectives were development and evaluation of a serum transfusion process in the thorny devil stick insect. Twenty-five clinically healthy adult insects (9.9-23.0 g) were enrolled and baseline body weights were collected. Hemolymph collection was not successful in males, thus, all were recipient only ( n = 12, MR). Females were divided into donor ( n = 4, FD) or donor and recipient ( n = 9, FDR) groups. Females were manually restrained and up to 1 mL of hemolymph was collected from the abdominal segment caudal to the proximal hindlimb using an 18 g hypodermic needle and passive collection via gravity. Hemolymph was quantified, centrifuged, and the serum separated. Insects were then injected superficially at the lateral aspect of the abdomen with 0.01 mL/g FD serum (MR), lactated Ringer's solution (LRS) equal to collected hemolymph volume (FD), or a combination of FDR serum and LRS equal to collected hemolymph volume (FDR). Response to stimulation, surface temperature, and righting reflex and mortality were serially assessed for up to 24 h and 7 days, respectively, following injection. In FD, median (range) injected LRS dose was 0.04 (0.03-0.06) mL/g. In FDR, median (range) injected serum, LRS, and combined serum and LRS dose was 0.03 (0.02-0.04), 0.01 (0-0.04), and 0.04 (0.02-0.06) mL/g, respectively. A mild temperature increase (maximum +2.9°C) (MR n = 10, FD n = 3, FDR n = 8) and delayed righting reflex (MR n = 4, FD n = 3, FDR n = 7) occurred in a subset of insects following injection. Two deaths occurred at 2 min ( n = 1, FDR) and 96 h ( n = 1, FD) post-injection. This is the first report of serum transfusions in thorny devil stick insects, and while largely successful, minor to severe transfusion reactions may occur., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Gregory, Heniff, Gorges, Lathan, Lewbart and Balko.)

The pink land iguana, Conolophus marthae, is one of four species of iguanas (three terrestrial and one marine) in the Galápagos Islands, and the only one listed as critically endangered by the IUCN. The species can only be found on the north-west slopes of the highest volcano on Isabela Island and was first described to science in 2009. As part of a population telemetry study, a health assessment was authorized by the Galápagos National Park. Wild adult iguanas were captured on Wolf Volcano in September 2019 and April 2021 to record morphological and physiological parameters including body temperature, heart rate, intraocular pressures, tear formation, and infrared iris images. Blood samples were also collected and analyzed. An i-STAT portable blood analyzer was used to obtain values for base excess in the extracellular fluid compartment (BEecf), glucose (Glu), hematocrit (HctPCV), hemoglobin (Hb), ionized calcium (iCa), partial pressure of carbon dioxide (pCO2), partial pressure of oxygen (pO2), percent oxygen saturation (sO2%), pH, potassium (K), and sodium (Na). When possible, data were compared to previously published and available data for the other Galápagos iguanas. The results reported here provide baseline values that will be useful in detecting changes in health status among pink land iguanas affected by climate change, invasive species, anthropogenic threats, or natural disturbances. The collected data also provide an invaluable resource for conservation scientists planning to implement conservation strategies, like translocations, that may temporarily alter these baseline values., Competing Interests: The authors have declared that no competing interests exist.

The moon jellyfish ( Aurelia aurita ) is a scyphozoan frequently maintained in public and private aquaria. Little research has been conducted to investigate the effects of various drugs, such as anesthetics, in this species. Tricaine methanesulfonate (MS-222), a common immersion anesthetic for fish and amphibians, was evaluated in a managed population of moon jellyfish. Twenty-four clinically healthy jellyfish were assigned into three groups of eight for trials of 0.3 g/L MS-222 (low concentration [LC]), 0.6 g/L MS-222 (high concentration [HC]), and a saltwater control. The goal was to evaluate the effects of MS-222 administration on moon jellyfish movement and response to stimuli. Movement and response to stimuli were measured via rocking and probe stimulus tests and observations of bell contraction quality and body tone. These tests were performed at baseline and throughout both drug exposure and recovery periods. A threshold drug effect was defined based on systematic scoring criteria. Additionally, elastomer tags were administered to four of eight animals in each MS-222 group to evaluate response to tag placement after drug exposure. Threshold drug effect was achieved in six of eight individuals in the LC group and eight of eight individuals in the HC group. The LC group had median threshold and recovery times of 12.2 and 10.1 min, respectively, while the HC group had median threshold and recovery times of 4.0 and 19.9 min, respectively. The HC group had significantly faster time to threshold drug effect ( P < 0.001) and longer recovery times ( P = 0.005) than the LC group. In both the LC and HC tagged group, three of four jellyfish had no reaction to tag placement. All animals recovered uneventfully, and there were no mortalities. MS-222 at 0.3 and 0.6 g/L decreased movement and response to stimuli in moon jellyfish.

The Turtle Rescue Team (TRT) at North Carolina State University College of Veterinary Medicine currently sees over 500 chelonian cases a year. Each turtle receives a thorough physical examination as the main evaluation of its health because further diagnostics are cost prohibitive in a donation-based facility. Thus, there is a need to further understand how a few economical diagnostics can evaluate the prognosis of chelonians. The objectives were to evaluate lactate, PCV, total solids, glucose, temperature, and HR as prognostic indicators in multiple chelonian species and to determine how the change in lactate concentrations over a 24 h period affects prognosis. Eastern box turtles ( Terrapene carolina carolina ), yellow-bellied sliders ( Trachemys scripta scripta ), river cooters ( Pseudemys concinna ), painted turtles ( Chrysemys picta ), and common snapping turtles ( Chelydra serpentina ) were evaluated because of prevalence of presentation to the TRT, differences in habitat, and differences in physiology. Hyperlactatemia, an increased lactate over 24 h when the initial lactate concentration was greater than 5 mM/L, and anemia were determined to be significant prognostic indicators of mortality for all turtles, and hypoglycemia was a significant prognostic indicator of mortality for male turtles ( P < 0.05).

Sea turtles are often restrained manually for brief periods during veterinary evaluation and care in rescue, rehabilitation, research, and aquarium settings. Blood gas values and lactate are routinely evaluated during triage of sea turtles, and lactate clearance is of prognostic significance in cold-stunned individuals. Although increases in blood lactate have been associated with muscle exertion, experimental forced submergence, trawl and pound net capture, and general anesthesia, changes in blood lactate associated with short periods of manual restraint have not been evaluated. Venous blood gas and lactate values were tested in 16 juvenile loggerhead sea turtles ( Caretta caretta ) before and after manual restraint for a 15-min routine veterinary examination. The agreement of blood lactate values between two point-of care analyzers (i-STAT and Lactate Plus) was also compared. Blood pH and bicarbonate (HCO 3 - ) decreased significantly ( P < 0.001), and partial pressure of carbon dioxide (pCO 2 ) increased significantly ( P < 0.0001) after 15 min. Lactate increased significantly between time points for both analyzers ( P < 0.0001). Linear regression analysis showed excellent correlation for lactate measurements obtained on both analyzers ( r = 0.998). The mean difference in lactate concentrations between the analyzers was statistically significant, indicating that the methods cannot be used interchangeably ( P < 0.0001). Deming regression and Bland-Altman plots identified a slight negative proportional bias for lactate measurement by the Lactate Plus compared with the i-STAT. These results suggest that clinicians should evaluate blood gas values and lactate at the beginning of health evaluations and interpret serial lactate values in sea turtles with caution, because even short periods of manual restraint can induce lactic acidosis and considerably influence these values.

Objective: To describe the anatomy of the brachial plexus in eastern box turtles (Terrapene carolina carolina), develop a blind perineural injection technique for brachial plexus blockade and evaluate the distribution of three volumes of new methylene blue dye for injection in cadavers., Study Design: Prospective, randomized, blinded cadaveric study., Animals: A total of 24 frozen-thawed box turtle cadavers; two turtles identified with shoulder injuries were subsequently excluded from the study. The remaining 22 turtles weighed 397 (190-581) g, median (range)., Methods: The brachial plexus and regional anatomy were identified by dissection of seven cadavers to determine anatomic landmarks for a perineural injection technique. This technique was tested by randomizing 15 cadavers into one of three groups to be injected bilaterally with one of three volumes (0.1, 0.2 or 0.3 mL) of methylene blue dye 1% aqueous solution. Investigators blinded to the assigned group dissected cadavers 15 minutes after injection and used staining of the four cervical spinal nerves (C5-C8; 25% for each nerve) to record a staining score of the brachial plexus (0-100%)., Results: Based on descriptions of the anatomy of the brachial plexus, an injection technique was designed. Injections of 0.1 mL methylene blue dye resulted in nine/10 injections with 100% nerve stained, and one/10 injection with 50% (two) nerves stained. All injections of 0.2 or 0.3 mL of methylene blue dye resulted in 100% nerves stained., Conclusions and Clinical Relevance: Perineural injection of the brachial plexus with 0.1, 0.2 or 0.3 mL methylene blue dye was successful in 29/30 injections in box turtle cadavers weighing 190-581 g. Further studies are needed to determine the minimum volume of injectate that can be successfully used for this technique, and to evaluate its application and efficacy in live turtles., (Copyright © 2021 Association of Veterinary Anaesthetists and American College of Veterinary Anesthesia and Analgesia. Published by Elsevier Ltd. All rights reserved.)

Ketorolac is a non-steroidal anti-inflammatory drug administered as an analgesic in humans. It has analgesic effects comparable to opioids but without adverse effects such as respiratory depression or restrictions because of controlled drug status. We designed this study to examine the potential of ketorolac as an analgesic for sea turtle rehabilitative medicine. Our objective was to determine the pharmacokinetics of a single 0.25 mg/kg intramuscular dose of ketorolac in a population of 16 captive-raised juvenile loggerhead sea turtles (Caretta caretta). A sparse sampling protocol was utilized, and blood samples were collected for 12 hours after administration of ketorolac. Samples were analyzed with high-pressure liquid chromatography (HPLC), and a nonlinear mixed effects model (NLME) was used to determine parameters for the population. With these methods, we identified a long elimination half-life (βT 1/2  = 11.867 hr) but a low maximum concentration (C MAX  = 0.508 µg/mL) and concentrations were below the level proposed to be therapeutic in humans (EC 50  = 0.1-0.3 μg/mL) for most of the collection period. We conclude that ketorolac may not be an appropriate long-term analgesic for use in loggerhead sea turtles at this dose; however, it may have some benefit as a short-term analgesic., (© 2021 John Wiley & Sons Ltd.)

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., Competing Interests: The authors declare there are no competing interests., (©2021 Dass et al.)

This study investigated the efficacy of isoflurane in Madagascar hissing cockroaches ( Gromphadorhina portentosa ). Cockroaches ( n = 12) were placed in a 2-L anesthesia chamber with isoflurane initiated at 5% at 2 L/m. Movement of antennae, mandibles, and legs and righting reflex were assessed, with full induction achieved when all responses had ceased for 2 m. Cockroach movement became ataxic on average (±SD) 3.5 ± 0.9 m after isoflurane treatment, and induction occurred on average 18.7 ± 4.4 m after treatment. Loss of righting reflex was the most consistent indication of full induction. Cockroaches fully recovered on average 16.2 ± 5.6 m after removal of isoflurane inhalant. Induction was uneventful, and no mortality or obvious morbidity was observed in treated cockroaches up to 30 d posttreatment. The authors conclude that the use of inhalant isoflurane is both safe and effective for anesthesia of Madagascar hissing cockroaches.

Island species are particularly vulnerable to environmental disturbances and introduced pathogens. Conducting health assessments of wild populations in the Galápagos improves the ability of wildlife managers and veterinarians to detect deteriorations in health status. Seabirds in particular are useful species to monitor due to their colonial breeding and wide migration range. Nazca boobies ( Sula granti ) in a breeding colony at Daphne Major ( n = 30) were given physical examinations, and blood samples were collected for hematology and biochemistry using an iSTAT Portable Clinical Analyzer. Female boobies had longer wing length than males, as well as lower blood urea nitrogen, creatinine, and white blood cell counts. This could be attributed to sexual dimorphism or differences in foraging and mating strategies between the sexes. The time between capture and blood collection had a significant inverse relationship on plasma sodium, potassium, hemoglobin, anion gaps, and lymphocyte counts, suggesting that blood sampling in Nazca boobies should be done in less than 5 m to avoid the impacts of stress on hematological parameters. This is the first health assessment on the breeding colony of Nazca boobies at Daphne Major, and the results can inform future monitoring in this species as well as other sulids.

Case Description: 4 wild adult rat snakes ( Pantherophis alleghaniensis ) were evaluated after ingesting spherical or ovoid foreign bodies., Clinical Findings: Physical examination revealed a large, firm mass at the level of the stomach in each snake. Radiographic findings were consistent with ingestion of a golf ball (3 snakes) or an artificial egg (1 snake). Signs consistent with prolonged impaction included scale loss, dermal abrasions, and apparent loss of body condition in one snake and regional skin ulceration, dehydration, and generalized muscle atrophy in another., Treatment and Outcome: Nonsurgical removal of the foreign body was attempted in anesthetized or heavily sedated snakes by external manipulation in the orad direction. A golf ball was removed through the oral cavity without complications in 1 snake. In the other 3 snakes, tension caused by the advancing foreign body resulted in full-thickness skin rupture in the cervical region. The procedure was completed with the use of a balloon catheter to aid foreign body advancement for 1 of the 3 snakes, and the skin defect was closed. The procedure was converted to esophagotomy for the other 2 snakes. Three snakes recovered and were released; 1 died of complications from prolonged impaction and esophageal perforation., Clinical Relevance: The described nonsurgical techniques for removal of ingested round or ovoid foreign bodies were associated with substantial complications in 3 of 4 treated rat snakes. Although a nonsurgical method for removal of ingested objects such as golf balls could benefit snakes, the methods used for these patients did not appear to be more beneficial than traditional gastrotomy.

Reference intervals of blood analytes are important diagnostic tools that clinicians use as a basis for making clinical decisions and that have been documented for many fish species. Although the effects of dietary supplementation on hematology and blood biochemistry variables in clownfish have been shown, there are no published reference intervals to date for routine hematology and blood biochemistry panels for these popular cultured marine fish (including Amphiprion spp.). The objective of this study was to establish de novo reference intervals for selected hemogram data and, using whole-blood analysis, commonly utilized chemistry analytes in two species of aquacultured clownfish: the Tomato Clownfish Amphiprion frenatus and Saddleback Clownfish A. polymnus. This study is based on blood samples from 25 clinically normal Tomato Clownfish and 38 clinically normal Saddleback Clownfish from two clownfish production facilities in Florida. Guidelines by the American Society for Veterinary Clinical Pathology were followed to generate robust reference intervals from the data collected. We report descriptive statistics and reference intervals for several hematology (n = 12) and biochemistry (n = 8) analytes using a small sample size and a small amount of blood per sample. The blood analyte data from this study are comparable with hematology and biochemistry data that have been reported in other teleost species. These results provide baseline health data for clownfish species that can be used in various clinical settings while also serving as a springboard for further research studies., (© 2020 American Fisheries Society.)

Invertebrate animals comprise more than 95% of the animal kingdom's species and approximately 40 separate phyla. Yet, invertebrates are an artificial taxon, in which all members simply possess a single negative trait: they lack a vertebral column (backbone). In fact, some invertebrates are more closely related to vertebrates than to their "fellow" invertebrates. For the purpose of this veterinary article, we have elected to review a handful of important groups: Coelenterates, Gastropods, Cephalopods, Chelicerates, Crustaceans, Insects, and Echinoderms. We have primarily included behaviors that may have an impact on clinical case outcome, or be of interest to the veterinary clinician., Competing Interests: Disclosure The authors have nothing to disclose., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Immersion in tricaine methanesulfonate (MS-222) is insufficient for euthanasia in at least one species of fish. The current study investigated the effectiveness of potassium chloride (KCl) to euthanize anesthetized koi ( Cyprinus carpio ). Twenty-eight healthy koi were anesthetized via immersion in 500 mg/L of buffered MS-222 for 10-12 min, manually removed to room air, and randomly administered 10 mEq/kg KCl (333 mg/ml) via one of three routes-intracardiac injection (IC) ( n = 7), intracoelomic injection (ICe) ( n = 7), or topical instillment over the gill filaments bilaterally (T) ( n = 7)-or received no treatment (control, C) ( n = 7). A Doppler ultrasonic flow detector was placed over the heart, and sounds were assessed continuously from immediately prior to treatment until 5 min posttreatment and every 5 min thereafter until Doppler sound cessation, resumption of operculation, or 30 min. Time to Doppler sound cessation or resumption of operculation was recorded. Doppler sound cessation occurred in 7/7 fish in IC (median 0.08, range 0-2.75 min) and 1/7 fish in T (10 min). In T, ICe, and C, 6/7, 7/7, and 7/7 fish, respectively, maintained Doppler sounds to 30 min. All fish in ICe (7/7) and C (7/7) resumed operculation with median (range) times of 22 (7-30) min and 16 (9-29) min from treatment, respectively. Intracardiac KCl at 10 mEq/kg rapidly ceases Doppler sounds and is a successful technique for euthanasia of anesthetized koi. Intracoelomic and topical KCl at 10 mEq/kg were not effective for euthanasia of koi.

The objective of this study was to document the pharmacokinetics of ketoprofen following 3 mg/kg intramuscular (IM) and intravenous (IV) injections in rainbow trout ( Oncorhynchus mykiss ) and 8 mg/kg intramuscular (IM) injection in Nile tilapia ( Oreochromis niloticus) . Plasma was collected laterally from the tail vein for drug analysis at various time intervals up to 72 h following the injection of ketoprofen. In trout, area under the curve (AUC) levels were 115.24 μg hr/mL for IM and 135.69 μg hr/mL for IV groups with a half-life of 4.40 and 3.91 h, respectively. In both trout and tilapia, there were detectable ketoprofen concentrations in most fish for 24 h post-injection. In tilapia, there was a large difference between the R- and S-enantiomers, suggesting either chiral inversion from R- to S-enantiomer or more rapid clearance of the R-enantiomer. AUC values of the S- and R-enantiomers were 510 and 194 μg hr/Ml, respectively, corresponding to a faster clearance for the R-enantiomer. This study shows that there were very high plasma concentrations of ketoprofen in trout and tilapia with no adverse effects observed. Future studies on the efficacy, frequency of dosing, analgesia, adverse effects, and route of administration are warranted., (Copyright © 2020 Greene, Mylniczenko, Storms, Burns, Lewbart, Byrd and Papich.)

The swallow-tailed gull, Creagrus furcatus , is a seabird endemic to the Galápagos archipelago. In general health, blood chemistry and haematology, parameters have not been published for this species. Blood analyses were run on samples drawn from 58 clinically healthy swallow-tailed gulls captured at Islote Pitt on San Cristóbal Island in July 2016 (28) and South Plaza Island in June 2017 (30). A point of care blood analyzer (iSTAT) was used in the field to obtain results for HCO 3 - , pH, pCO 2 , pO 2 , TCO 2 , anion gap, chloride, creatinine, glucose, haematocrit, haemoglobin, ionized calcium, potassium, sodium and urea nitrogen. A portable Lactate Plus™ analyzer was used to measure lactate. The baseline data reported is valuable for comparisons amongst different populations in the archipelago and to detect changes in health status of Galápagos swallow-tailed gulls., (© The Author(s) 2020. Published by Oxford University Press and the Society for Experimental Biology.)

Significant impacts of global climate change on wildlife have been documented and are projected to continue. Reptiles have been suggested as being especially susceptible to these effects along with other anthropogenic impacts on their environment. A retrospective review of medical records for 2,356 wild eastern box turtles ( Terrapene carolina carolina ) admitted to the Turtle Rescue Team (TRT) at the North Carolina State University College of Veterinary Medicine between 1996 and 2017 was performed in order to: 1) report common presenting complaints, 2) describe the timeline of when box turtles were admitted to the TRT for each year of the study, and 3) investigate temporal shifts in turtle admissions and associations with changes in environmental temperature over a 22-yr period. The most common presenting complaint was vehicular trauma ( n = 1,195) with over 70% of the caseload associated with anthropogenic impacts. Average monthly temperatures from 1996 to 2017 for Raleigh, North Carolina, USA, were extracted from the National Oceanic and Atmospheric Administration climate database. By comparing the pre- and post-2006 years using the Mann-Whitney test, we found that both the annual peak temperature and the annual timing of admission to the TRT have shifted significantly or with a strong tendency towards significance ( P = 0.0008 and 0.052, respectively). Annual peak temperature has increased by 1.3°C, and timing of admission has shifted 18 days earlier between pre- and post-2006 years. This supports the hypothesis that box turtle activity patterns are shifting and that these changes are potentially related to climate.

A healthy adult, intact female keeled box turtle (Cuora mouhotii) was found to have a marked heterophilic leukocytosis using normal hematologic parameters established for the eastern box turtle (Terrapene carolina carolina), a related chelonian species. This animal was monitored with serial complete blood counts (CBCs) over the next 15 years despite remaining asymptomatic for an infectious condition. Retrospective CBC data were compiled from 38 presumably healthy keeled box turtles to establish hematologic values for comparison in this species. Using this species-specific data, over the 15-year period, the female keeled box turtle had two times where the white blood cell (WBC) count was greater than 2 standard deviations (SD) above the mean, six times where the WBC count was greater than 1 SD above the mean, six times where the PCV was greater than 2 SD above the mean, and eight times where the PCV was greater than 1 SD above the mean. Infection and inflammation are the most common causes of leukocytosis in reptiles; however, given the clinical presentation of this patient, it was postulated that these clinicopathologic changes could be secondary to a stress response. Establishing reference intervals and understanding how stress impacts CBC parameters are important for evaluating the health status of keeled box turtles kept in captivity and for assessing the effects of environmental changes on the health status of wild populations of this endangered chelonian species., (© 2020 American Society for Veterinary Clinical Pathology.)

Lepidobatrachus laevis , commonly called the Budgett frog, is a member of the horned frog family (Ceratophryidae), which has become increasingly popular among amphibian hobbyists. L. laevis is also used in biologic research on embryonic development, providing a novel model species for the study of organogenesis, regeneration, evolution, and biologic scaling. However, little scientific literature details disease processes or histologic lesions in this species. Our objective was to describe spontaneous pathologic lesions in L. laevis to identify disease phenotypes. We performed a retrospective analysis of 14 captive L. laevis frogs (wild-caught and captive-bred), necropsied at the NC State University College of Veterinary Medicine between 2008 and 2018. The majority of frogs exhibited renal changes, including varying combinations of tubular epithelial binucleation, karyomegaly, and cytoplasmic vacuolation; polycystic kidney disease; and renal carcinoma. Many of the renal changes are reminiscent of a condition described in Japanese ( Bufo japonicus ) and Chinese (Bufo raddei) toad hybrids that progresses from tubular epithelial atypia and tubular dilation to polycystic kidney disease to renal carcinoma. A second common finding was variably sized, randomly distributed bile duct clusters (biliary proliferation). Other noteworthy findings included regional or generalized edema, intestinal adenocarcinoma, aspiration pneumonia, and parasitism. This retrospective analysis is the first description of histologic lesions identified in captive L. laevis populations, providing new insight into spontaneous disease processes occurring in this species for use in disease diagnosis and clinical management.

Enrofloxacin is a fluoroquinolone widely used in animals including fish. Intramuscular (IM) injection of enrofloxacin is a feasible and efficacious option for drug delivery. In many species IM injection has been associated with injection site reactions and increases in serum muscle enzymes. Injection site reactions have not been well characterized in fish. Three groups of striped bass ( Morone saxatilis ) received an IM injection of enrofloxacin 2.27% in the right epaxial musculature 24, 48, or 96 hr prior to evaluation. Mean dose was 7.69 mg/ kg (6.14-9.69 mg/kg). The 24- and 48-hr groups received an injection of equal-volume 0.9% saline in the left epaxial musculature. A corresponding noninjected tissue sample was designated in the left epaxial musculature from each fish of the 96-hr group. Fish were euthanized and injection sites and noninjection control sites were evaluated grossly and histologically. Grades 1-4 were assigned to samples, with grade 1 corresponding to normal tissue and grades 2, 3, and 4 corresponding to mild, moderate, and severe inflammation and/or necrosis respectively. Externally, all control and injection sites appeared visually unremarkable. On cut surface, epaxial muscle of the enrofloxacin-injected tissue appeared moderately to severely hemorrhagic compared to saline and noninjected tissue, which was normal or mildly hemorrhagic. Histologically, eight of eight noninjected tissues were grade 1. For saline-injected tissues, 14 of 16 tissues were grade 2 and 2 samples were grade 3 when 24- and 48-hr groups were combined. For enrofloxacin-injected tissues, 8 of the 8 24-hr samples were grade 3 and 16 of the 16 48- and 96-hr samples were grade 4. These data show that IM injection of enrofloxacin 2.27% is associated with severe hemorrhage, necrosis, and inflammation in striped bass, and may negatively affect animal welfare.

Enrofloxacin is frequently administered to turtles in wildlife clinics during rehabilitation due to its wide spectrum of antibacterial activity and availability of injectable formulations. However, sufficient pharmacokinetic data to guide dosing are lacking. The objective of this study was to determine pharmacokinetic parameters of enrofloxacin and its active metabolite, ciprofloxacin, in chelonians presenting injured to a wildlife clinic. Thirty-six Eastern box turtles (EBT, Terrapene carolina carolina), 23 yellow-bellied sliders (YBS, Trachemys scripta scripta), and 13 river cooters (RC, Pseudemys concinna) received a single subcutaneous injection of enrofloxacin at 10 mg/kg. Blood samples were collected between 0 and 240 hr postinjection. Pharmacokinetic parameters were determined using nonlinear mixed-effects modeling (NMLE). Overall elimination half-life (T½) was over 75 hr, and varied among species. T½ was 63 hr in EBT and 79 hr in YBS, which is longer than in previous reports. The volume of distribution (steady-state) was 1.4 L/kg across turtle species, but highly variable-ranging from 0.4 L/kg in RC to 1.9 L/kg in YBS. Antibiotic concentrations were above a minimum inhibitory concentration value of 0.5 µg/ml for over 200 hr. These results indicate variable pharmacokinetic parameters for enrofloxacin among turtle species, which will help guide appropriate dosing protocols in injured turtles., (© 2020 John Wiley & Sons Ltd.)