Olimpia R. Lai

AFLATOXINS IN AQUATIC SPECIES: METABOLISM, TOXICITY AND PERSPECTIVES

Among all known mycotoxins, aflatoxins represent the most investigated, widespread and worrisome source of contamination of foods and feed worldwide. In the early 1960s, soon after the finding of aflatoxin B1 (AFB1) in the feedstuffs of aquacultured rainbow trout that had died in an epizootic of hepatomas, great scientific discoveries were made in several areas by a number of researchers under the direction of scientists like J. Halver, R. 0. Sinnhuber, G. S. Bailey, J. D. Hendricks and colleagues. Since that time, several studies have focused on the identification of new isoenzymes involved in AFB1 metabolism and on the discovery of new modulators in AFB1-induced cancer initiation and progression. However, metabolic and toxicological studies on aflatoxins in marine aquacultured species are fragmented and restricted to a limited number of fish species. Aflatoxins exert a substantial impact on the fish farming production, causing disease with high mortality and a gradual decline of reared fish stock quality, thus representing a significant problem in aquaculture systems. Based on these considerations, the goals of this review article are: (1) to gather the currently available scientific information, summarising existing data on aflatoxin contamination on feeds and fishmeals, and toxicological effects induced in reared aquatic species; (2) to make a comparative analysis of AFB1 metabolism in the most representative species studied; (3) to gain new insights on the risk of DNA damage caused by aflatoxins on fish genomes and their role in cancer development.

Pharmacokinetics of imidocarb dipropionate in horses after intramuscular administration

The objective of this study was to determine the pharmacokinetic behaviour of imidocarb in horses following a single i.m. injection at the dose commonly administered to treat Babesia caballi infections or to prevent babesiosis. Eight horses were injected i.m. with a single dose of 2.4 mg imidocarb dipropionate/kg bwt and blood, faecal, urine and milk samples were collected. For imidocarb determination, a high-performance liquid chromatographic method (HPLC) was used after weak cation-exchange solid phase, or liquid-liquid, extraction procedures. Twelve hours after treatment, no detectable plasma concentrations were recorded in any of the treated animals. The distribution and elimination patterns of the drug suggested that it is quickly sequestrated in some storage tissues and remains in the body for a long time. Its prolonged presence in the body may confer a reservoir effect to imidocarb in some tissues, therefore making it undetectable in the plasma of animals but sufficient to produce its described therapeutic and prophylactic activities.

Pharmacokinetic Behavior of Enrofloxacin in Estuarine Crocodile (Crocodylus porosus) after Single Intravenous, Intramuscular, and Oral Doses

Abstract The disposition kinetics of enrofloxacin at a single dose of 5 mg/kg body weight were determined in clinically healthy captive-reared estuarine crocodiles (Crocodylus porosus) after intravenous (i.v.), intramuscular (i.m.), and oral (p.o.) administration. Enrofloxacin plasma concentrations were determined by diode array detection–high-performance liquid chromatography (limit of detection/limit of quantitation: 0.05 µg/ml). Data were subjected to noncompartmental analysis. The integrated pharmacokinetic-pharmacodynamic (PK-PD) variables showed that optimal area under the curve from the time of dosing to 24 hr:minimal inhibitory concentration (MIC) (>125) and peak plasma concentrations:MIC (>8) ratios, as reported for concentration-dependent bactericidal antimicrobials like fluoroquinolones, were achievable with both a single i.v. or i.m. dose for susceptible microorganisms with MIC values of ≤0.5 µg/ml, while the relatively slow onset of peak time allowed an effective plasma drug level only on day 3. The persistence of useful plasma concentrations indicated the possibility of redosing every 3 day for parenteral routes of administration, while further studies are needed for the oral route. Nevertheless, the absence of adverse reactions in the animals following i.v., i.m., or p.o. administration of enrofloxacin after a single dose of 5 mg/kg indicates the possibility of its safe and effective clinical use in captive estuarine crocodiles.

Pharmacokinetic Behavior of Meloxicam in Loggerhead Sea Turtles (Caretta caretta) after Intramuscular and Intravenous Administration

Abstract Data on reptile analgesia are scarce for nonsteroidal anti-inflammatory drugs (NSAIDs) and opioids and almost completely lacking in sea turtles, even though emergencies requiring correct pain management are very frequent in their rehabilitative medicine; therefore, dosage regimens extrapolated from other species involve the risk of clinical failure and damage to the animals. We describe the pharmacokinetic behavior of meloxicam in the loggerhead sea turtle (Caretta caretta). We chose meloxicam because of its selective anti-cyclooxygenase-2 activity and lesser adverse side effects. No data are available on the capacity of turtles to tolerate NSAIDs, so we chose a dose of 0.1 mg/kg of meloxicam. Plasma concentrations of meloxicam were unexpectedly low both for intravenous (IV; maximum concentration [Cmax] = 0.04±0.02 µg/mL) and intramuscular (IM; Cmax = 0.07±0.09 µg/mL) administration. A double-peak phenomenon occurred after both IV (time for second peak concentration Tmax2 = 10.33±10.89 h) and IM (Tmax2 = 1.17±0.75 h). The second peak after IM injection was premature, so some difficulty and delay in absorption appears to be an appropriate explanation. Furthermore, the area under the curve, and therefore systemic bioavailability (F = 31.82±28.24%), after both IV (0.30±0.29) and IM (0.10±0.03) injection appeared particularly limited. Terminal elimination slope and mean residence time indicated fast elimination after IM dosing; as a consequence, plasma concentrations dropped below analytic limits in 8 h. Considering that IM is the favored route of administration of drugs in rescue centers, it is unlikely that meloxicam at 0.1 mg/kg is an appropriate choice, particularly in long-term pain management protocols.

Pharmacokinetics of marbofloxacin after a single oral dose to loggerhead sea turtles (Caretta caretta).

The single-dose disposition kinetics of marbofloxacin (MBX) were determined in clinically healthy loggerhead sea turtles (n=5) after oral (PO) administration of 2 mg kg(-1) bodyweight. Marbofloxacin plasma concentrations were determined by DAD-HPLC (LOD/LOQ 0.015/0.05 microg ml(-1)). Data were subjected to non-compartmental analysis. Following PO administration, marbofloxacin achieved maximum plasma concentrations of 11.66+/-2.53 mg L(-1) at 15.00+/-3.00 h. The absence of general adverse reactions in the turtles of the study, and the favourable pharmacokinetic properties (long half-life and high maximum plasma concentration) of MBX administered PO at the single-dose of 2 mg kg(-1) suggest the possibility of its safe and effective clinical use in loggerhead sea turtles.

Pharmacokinetics of Marbofloxacin in Loggerhead Sea Turtles (Caretta caretta) after Single Intravenous and Intramuscular Doses

Abstract The disposition kinetics of marbofloxacin at a single dose of 2 mg/kg bodyweight were determined in a crossover trial with five clinically healthy loggerhead sea turtles (Caretta caretta) after i.v. and i.m. administration. Marbofloxacin plasma concentrations were determined by high-performance liquid chromatography (LOD/LOQ 0.05 µg/ml). Data were subjected to noncompartmental analysis. The integrated pharmacokinetic/pharmacodynamic variables showed that optimal area under the curve (AUC0–24 h): minimal inhibitory concentration (MIC) (>125) and Cmax: MIC (>8) ratios, as reported for concentration-dependent bactericidal antimicrobials (e.g., fluoroquinolones), were achievable with both a once daily i.v. or i.m. dose for microorganisms with MIC ≤ 0.5 µg/ml, while a Cmax: MIC > 8 for MIC ≥ 1 µg/ml was achievable only after the i.v. administration. The absence of adverse reactions in the animals after i.v. or i.m. administration of marbofloxacin and the favorable pharmacokinetic/pharmacodynamic properties after a single dose of 2 mg/kg suggest the possibility of its safe and effective clinical use in loggerhead sea turtles.

Surgical treatment of injuries caused by fishing gear in the intracoelomic digestive tract of sea turtles

We report the surgical techniques used to remove accidentally ingested hooks and branchlines localized in different parts of the digestive tract of 129 loggerhead sea turtles Caretta caretta, together with the characteristics and localization of lesions, and final outcome related to their severity. Hooks were removed from the cervical esophagus via the ventral surface of the neck, while the supraplastron approach was performed for hooks wedged in the intracoelomic portion of the esophagus. An approach through the left axillary region was preferred for fishhooks in the stomach, while hooks and long branchlines in the intestine or pyloric area were removed by approaching the coelomic cavity through the right or left prefemoral fossa. The ingestion of fishhooks, and/or longlines, often induces severe injuries in the digestive tract that could lead to the death of the turtles, with the extent of damage engendered by lines often more severe than that caused by hooks, leading to strangulation, intussusception, and tears that require resection of long tracts of intestine. Spontaneous expulsion of hooks, even where possible, involves long waiting times, with the possible impairment of the turtles clinical condition, and should be avoided when the line is evident or suspected. The development of diversified surgical techniques enabled us to approach the coelomic cavity with minimally invasive and easy-to-perform methods, and survival rates proved very satisfactory.

DIAGNOSIS AND TREATMENT CONSIDERATIONS IN A CASE OF MALIGNANT MESENCHYMOMA IN AN AFRICAN FUR SEAL (ARCTOCEPHALUS PUSILLUS)

Abstract: A 20-yr-old African fur seal (Arctocephalus pusillus) presented with a slowly growing mass located on the dorsum at the level of the last thoracic vertebrae. The mass was hard, 10 cm in diameter, and not adherent to the underlying tissues. Multiple biopsies were collected for histopathology and revealed extensive areas of necrosis, small nodules of malignant mesenchymal proliferation with areas of chondroid metaplasia, and atypical cells in vessel walls. The morphologic diagnosis was suggestive of malignant mesenchymal neoplasia originating from the vascular wall. The mass was removed 1 mo later due to ulceration and infection. Histologically, based on the World Health Organizations classification of neoplastic processes in domestic animals, the tumor was consistent with malignant mesenchymoma. The margins of resection revealed the presence of neoplastic cells. Based on these results, the particular species involved, the high local invasiveness, and the high metastatic index of this malignant tumor in domestic mammals and humans, the prognosis was poor. The animal died 6 mo later with metatastic disease.

Management of severe head injury with brain exposure in three loggerhead sea turtles Caretta caretta

The loggerhead Caretta caretta is the most common sea turtle in the Mediterranean. Currently, sea turtles are considered endangered, mainly due to the impact of human activities. Among traumatic lesions, those involving the skull, if complicated by brain exposure, are often life-threatening. In these cases, death could be the outcome of direct trauma of the cerebral tissue or of secondary meningoencephalitis. This uncontrolled study aims to evaluate the use of a plant-derived dressing (1 Primary Wound Dressing®) in 3 sea turtles with severe lesions of the skull exposing the brain. Following surgical curettage, the treatment protocol involved exclusive use of the plant-derived dressing applied on the wound surface as the primary dressing, daily for the first month and then every other day until the end of treatment. The wound and peri-wound skin were covered with a simple secondary dressing without any active compound (non-woven gauze with petroleum jelly). Data presented herein show an excellent healing process in all 3 cases and no side effects due to contact of the medication with the cerebral tissue.