BLOOD CHEMISTRY AND HEMATOLOGY VALUES IN HEALTHY AND REHABILITATED ROUGH-TOOTHED DOLPHINS ( STENO BREDANENSIS).

Rehabilitation efforts for live stranded marine mammals are guided by diagnostic measures of blood chemistry and hematology parameters obtained from each individual undergoing treatment. Despite the widespread use of blood parameters, reference values are not available in the literature from healthy rough-toothed dolphins ( Steno bredanensis) with which to infer the health status of an animal. We examined serum or plasma chemistry and hematology data from 17 rough-toothed dolphins either housed at Dolphin Quest French Polynesia or during their rehabilitation at the Dolphin and Whale Hospital in Sarasota, Florida, US between 1994 and 2005. Blood parameters were compared among healthy animals, rehabilitation animals that were eventually released, and rehabilitation animals that died. This study indicated significant differences in many blood parameters for the poorly known rough-toothed dolphin that are likely to vary between healthy and sick animals. These included aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase, bicarbonate, and globulins, which were greater in sick dolphins, and alkaline phosphatase and total protein which were greater in healthy individuals. Total white blood cell counts were lower in healthy animals as were the absolute numbers of neutrophils, monocytes, and eosinophils. Analysis of first blood sample levels for glucose, sodium, and erythrocyte sedimentation rate may have value for triage and prognostic evaluation.

Aerobic microorganisms associated with free-ranging bottlenose dolphins in coastal Gulf of Mexico and Atlantic Ocean waters.

Our abilities to assess health risks to free-ranging dolphin populations, to treat live-stranded or captive dolphins, and to evaluate the risks of disease transmission between humans and dolphins have suffered from a lack of basic information on microorganisms associated with normal, presumably healthy free-ranging individuals. In order to provide these data, we sampled free-ranging bottlenose dolphins (Tursiops truncatus) off Florida, Texas, and North Carolina during 1990-2002. Blowhole and anal/fecal samples yielded 1,871 bacteria and yeast isolates and included 85 different species or groups of organisms. Vibrios, unidentified pseudomonads, Escherichia coli, Staphylococcus spp., and a large group of nonfermenting gram-negative bacteria represented >50% of isolates. Vibrio alginolyticus and Vibrio damsela were the most commonly recovered bacteria from both anal/fecal and blowhole samples. Many organisms occurred sporadically in dolphins that were sampled repeatedly, but some were consistently isolated from individual animals and may indicate the carrier state. Vibrios were common, but some geographic variability in the presence of these and other organisms was noted. Potential pathogens of significance to humans and other animals were recovered.

Integrating life-history and reproductive success data to examine potential relationships with organochlorine compounds for bottlenose dolphins (Tursiops truncatus) in Sarasota Bay, Florida.

Research initiated in 1970 has identified a long-term, year-round resident community of about 140 bottlenose dolphins (Tursiops truncatus) in Sarasota Bay, Florida, providing unparalleled opportunities to investigate relationships between organochlorine contaminant residues and life-history and reproductive parameters. Many individual dolphins are identifiable and of known age, sex, and maternal lineage (< or =4 generations). Observational monitoring provides data on dolphin spatial and temporal occurrence, births and fates of calves, and birth-order. Capture-release operations conducted for veterinary examinations provide biological data and samples for life-history and contaminant residue measurement. Organochlorine concentrations in blubber and blood (plasma) can be examined relative to age, sex, lipid content, and birth-order. Reproductive success is evaluated through tracking of individual female lifetime calving success. For the current study, 47 blubber samples collected during June 2000 and 2001 were analyzed for PCB concentrations of 22 congeners relative to life-history factors and reproductive success. Prior to sexual maturity, males and females exhibited similar concentrations of about 15-50 ppm. Classical patterns of accumulation with age were identified in males, but not in females. Subsequently, males accumulated higher concentrations of PCBs through their lives (>100 ppm), whereas females begin to depurate with their first calf, reaching a balance between contaminant intake and lactational loss (<15 ppm). In primiparous females, PCB concentrations in blubber and plasma and the rates of first-born calf mortality were both high. First-born calves had higher concentrations than subsequent calves of similar age (>25 vs.<25 ppm). Maternal burdens were lower early in lactation and increased as calves approached nutritional independence. Empirical data were generally consistent with a published theoretical risk assessment and supported the need for incorporation of threats from indirect anthropogenic impacts such as environmental pollutants into species management plans. Long-term observational monitoring and periodic biological sampling provide a powerful, non-lethal approach to understanding relationships between organochlorine residue concentrations in tissues and reproductive parameters for coastal dolphins.

Pharmacokinetics of ticarcillin in the loggerhead sea turtle (Caretta caretta) after single intravenous and intramuscular injections.

Three captive loggerhead sea turtles, Caretta caretta, were used in four trials, one i.v. and three i.m., to determine the pharmacokinetic properties of a single dose of ticarcillin. For the i.v. study, each turtle received a single 50 mg/kg dose and blood samples were collected at 0, 0.5, 1, 2, 4, 6, 8, and 12 hr and at 1, 1.5, 2, 2.5, 3, 4, 6, 8, 10, and 14 days after administration. For the i.m. study, each turtle received one of three dosages (25, 50, or 100 mg/kg) in a randomized complete block design and blood samples were collected at the same time intervals. Each trial was separated by a minimum of 28 days to allow for complete drug clearance. Drug concentration in plasma was determined by a validated liquid chromatography-mass spectrometry assay. For the i.v. study, the elimination half-life was 5.0 hr. The apparent volume of distribution and plasma clearance were 0.17 L/kg and 0.0218 L/hr/kg, respectively. For the i.m. study, mean time to maximum plasma concentrations ranged from 1.7 ( +/- 0.58) hr in the 50 mg/kg group to 3.7 (+/- 2.5) hr in the 100 mg/kg group. Mean bioavailability ranged from 45% ( +/- 15%) in the 50 mg/kg group to 58% (+/- 12%) in the 100 mg/kg group, and the mean residence time ranged from 7.5 ( +/- 2.6) hr in the 25 mg/kg group to 16 (+/- 6.8) hr in the 100 mg/kg group. Two turtles had slight alanine aminotransferase elevations that were not clinically apparent at two different dosages, but otherwise, blood chemistries were unaffected. Possible i.m. dosage regimens for loggerhead sea turtles are 50 mg/kg q24 hr or 100 mg/kg q48 hr. Liver enzymes should be monitored during treatment.

Subacute atropine toxicity in a pygmy sperm whale, Kogia breviceps.

Atropine, an anticholinergic agent commonly used in human and veterinary medicine, is reported to cause toxicity associated with its antimuscarinic action. A juvenile pygmy sperm whale, Kogia breviceps, was treated with atropine in an attempt to relieve symptoms similar to pyloric stenosis, as has been used in humans. Two doses of 0.01 mg/kg were given i.m., 12 hr apart, followed by three doses of 0.005 mg/kg i.m. s.i.d. over the next 3 days. Symptoms associated with atropine toxicity developed gradually and included hyperexcitability, a generalized ascending paralysis of body musculature, shallow, rapid respiration, vomiting, aspiration of seawater, and pulmonary edema. Treatment with physostigmine salicylate (two doses of 2 mg i.m., I hr apart) was effective in counteracting the paralysis, as well as other symptoms, beginning in as little as 17 min after the first dose, and the whale was back to swimming on its own after 8 hr. All overt symptoms of atropine toxicity were gone in about 24 hr, but there were other possible sequella that lasted much longer.

Steady-state plasma concentrations of itraconazole after oral administration in Kemp's ridley sea turtles, Lepidochelys kempi.

Pharmacokinetic studies of antifungal agents in reptiles are uncommon. Itraconazole, which has been used prophylactically in juvenile sea turtles suffering from hypothermia (cold stunning) on a regular basis, was evaluated for steady-state plasma concentrations. Five Kemp's ridley sea turtles (Lepidochelys kempi) receiving itraconazole at several dosages in a rehabilitation program had blood collected within 24 hr to estimate dosing frequency. Subsequently, serial blood samples of Kemp's ridley sea turtles that were given itraconazole at several dosages for 30 days to treat cold stunning were collected at various intervals to evaluate itraconazole plasma concentrations. Tissue samples were collected from one Kemp's ridley that died during rehabilitation. Plasma concentrations of itraconazole (and of hydroxyitraconazole [OH-ITRA], one of its major bioactive metabolites) were determined using a modified, validated reverse-phase high-performance liquid chromatography technique. Itraconazole concentrations in tissues were determined by bioassay to be far greater than the plasma concentrations measured in any of the turtles. At a 15-mg/kg dosage, the half-life (t1/2) was 75 hr for itraconazole and 55 hr for OH-ITRA. All dosages produced adequate concentrations in some turtles, but consistent therapeutic concentrations were produced only at 15 mg/kg q72hr and 5 mg/kg s.i.d., with the latter producing the highest plasma concentrations.

Annual, seasonal and individual variation in hematology and clinical blood chemistry profiles in bottlenose dolphins (Tursiops truncatus) from Sarasota Bay, Florida.

Hematology and clinical blood chemistry (HCBC) profiles in free-living bottlenose dolphins from Sarasota Bay, Florida have been monitored over a 14-year period. This long-term dataset includes samples from recaptured dolphins, enabling individual variation to be accounted for when investigating seasonal and annual variability. Four different laboratories carried out the assays and inter-laboratory comparisons found significant differences in 31 of 39 parameters measured. However, variability in comparable HCBCs by sex, age, condition, season and year could be investigated. Significant relationships with the independent variables were found for the majority of the HCBCs. Notable consistent seasonal differences included significantly elevated glucose and significantly lower creatinine concentrations in winter compared to summer. These differences may be due to energetic or thermoregulatory fluctuations in the animals by season and do not necessarily have any clinical significance. Erythrocyte counts were significantly lower in the winter, possibly also due to nutritional differences. Albumin and calcium levels in this population have increased significantly over the years of monitoring and consistently across seasons, being higher in the winter than the summer. Again, nutritional and thermal constraints seem to be the most likely environmental factors influencing these patterns.

Comparison of diagnostic techniques for Helicobacter cetorum infection in wild Atlantic bottlenose dolphins (Tursiops truncatus).

Helicobacter cetorum sp. nov. has been cultured from the stomach of Atlantic white-sided dolphins (Lagenorhynchus acutus) and the feces of Pacific white-sided (L. obliquidens) and Atlantic bottlenose (Tursiops truncatus) dolphins and a beluga whale (Delphinapterus leucas). H. cetorum has high homology to Helicobacter pylori as shown by 16S rRNA sequencing, and H. cetorum infection has been associated with gastritis and clinical signs in cetaceans. Because the prevalence of H. cetorum in wild populations is unknown, minimally invasive techniques for detecting H. cetorum were compared for 20 wild bottlenose dolphins sampled as part of a long-term health study. Fecal samples were tested for helicobacter by culture, Southern blotting, and PCR using genus-specific and H. cetorum-specific primers. An enzyme-linked immunosorbent assay (ELISA) was developed to measure H. cetorum immunoglobulin G (IgG). H. cetorum was cultured from 4 of 20 fecal samples, 7 samples were positive using Helicobacter sp. PCR, and 8 samples were positive for H. cetorum using species-specific primers. Two additional fecal samples were positive by Helicobacter sp. Southern blotting, suggesting infection with another helicobacter. All 20 sera contained high levels of IgG antibodies to H. cetorum that were significantly lowered by preabsorption of the sera with whole-cell suspensions of H. cetorum (P < 0.02). Until the specificity of the serum ELISA can be determined by testing sera from dolphins confirmed to be uninfected, PCR and Southern blot screenings of feces are the most sensitive techniques for detection of H. cetorum, and results indicate there is at least a 50% prevalence of H. cetorum infection in these dolphins.

Disseminated mycotic infection caused by Colletotrichum acutatum in a Kemp's ridley sea turtle (Lepidochelys kempi).

Colletotrichum acutatum is a cosmopolitan plant pathogen with a wide host range. While the organism's phytopathogenic potential has been well documented, it has never been reported as an etiologic agent of disease in either animals or humans. In this case, a juvenile Kemp's ridley sea turtle, Lepidochelys kempi, probably with immune compromise following cold stunning (extended hypothermia), developed a disseminated mycotic infection in the lungs and kidneys. Prophylactic treatment with oral itraconazole did not prevent or cure the infection. This report of a Colletotrichum acutatum infection in an animal extends the range of disease caused by this organism beyond that of a phytopathogen.