M. Cristina Fossi

The use of non-destructive biomarker in Mediterranean cetaceans: Preliminary data on MFO activity in skin biopsy

Abstract Cetaceans have been subjected to heavy anthropogenic pressure in the last century. In addition to hunting there is now the subtle threat of pollution which may be responsible for metabolic impairment and involved in stranding episodes and population decline. The need to study pollutants and their effect on cetacean populations is often in contrast with the need to protect these animals. In this paper a new method of collecting skin and hypodermic biopsies in the Fin whale (Balaenoptera physalus) and Striped dolphin (Stenella coeruleoalba) in order to analyse Mixed Function Oxidase (MFO) activity and organochlorine contents, is described. The results show marked differences between the two species. MFO activity in skin and organochlorines in blubber are higher in Striped dolphin than the Fin whale. The new method enables the main effects of exposure to lipophilic contaminants to be assessed in a nondestructive way in biomarker studies.

The use of non destructive biomarkers in the study of marine mammals.

Marine mammals have been subject to heavy anthropogenic pressure by direct killing and chemical pollution all over the world. Most studies of contamination and biomarker responses in marine mammals have been conducted using animals killed by hunting out of a total of 12 cetacean species studied, 45 of the specimens were obtained by sacrificing the animal; out of a total of eight pinniped species studied, 40 of the specimens were obtained by killing. The development of a series of non destructive techniques to evaluate biomarker responses and residue levels is recommended for the hazard assessment and conservation of endangered species of marine mammals. Here we review the current status of the non destructive biomarker approach in marine mammals, describing the biological materials available for non destructive tests in stranded brain, liver, blood, skin, subcutaneous blubber, muscle and fur and free ranging animals blood, skin biopsy, fur and faeces and the respective biomarker techniques mixed function oxidase activity and DNA damage in skin biopsy samples; porphyrins in faeces and fur; esterases, porphyrins, clinical biochemical parameter, vitamin A and micronuclei in blood samples. Residue analysis can be carried out in the various biological materials. We also report the results of applying this methodological approach to cetaceans minke whale Balaenoptera acutorostrata, fin whale-- Balaenoptera physalus, beluga whale-- Delphinapterus leucas, short finned pilot whale-- Globicephala macrorhynchus, harbour porpoise -- Phocoena phocoena, Rissos dolphin-- Risso s Grampus griseus, Dall s porpoise-- Phocoenoides dalli dalli, melon headed whale-- Peponocephala electra, bottlenose dolphin -- Tursiops truncatus, striped dolphin-- Stenella coeruleoalba, spinner dolphin-- Stenella longirostris, killer whale-- Orcinus orca and pinnipeds northern fur seal- Callorhinus ursinus, hooded seal-- Cystophora cristata, grey seal-- Halichoerus grypus, harbour seal-- Phoca vitulina, ringed seal-- Phoca hispida, harp seal-- Phoca groenlandica, ribbon seal-- Phoca fasciata, largha seal- Phoca largha, southern sea lion-- Otaria flavescens in field studies for prognostic and diagnostic purposes.Marine mammals have been subject to heavy anthropogenic pressure by direct killing and chemical pollution all over the world. Most studies of contamination and biomarker responses in marine mammals have been conducted using animals killed by hunting out of a total of 12 cetacean species studied, 45 of the specimens were obtained by sacrificing the animal; out of a total of eight pinniped species studied, 40 of the specimens were obtained by killing. The development of a series of non destructive techniques to evaluate biomarker responses and residue levels is recommended for the hazard assessment and conservation of endangered species of marine mammals. Here we review the current status of the non destructive biomarker approach in marine mammals, describing the biological materials available for non destructive tests in stranded brain, liver, blood, skin, subcutaneous blubber, muscle and fur and free ranging animals blood, skin biopsy, fur and faeces and the respective biomarker techniques mixed function ...

Serum esterase inhibition in birds: A nondestructive biomarker to assess organophosphorus and carbamate contamination

With the aim of proposing a nondestructive biomarker for monitoring the toxicological risk to birds of exposure to the organophosphorus insecticide azamethiphos and the carbamate insecticide methomyl, laboratory studies were performed on serum “B” esterases in Japanese quail (Coturnix coturnix japonica). The birds received two single dose treatments of each compound (azamethiphos and methomyl), i.e., 50 mg/kg and 250 mg/kg respectively. In the first treatment, serum butyrylcholinesterase (BChE) and carboxylesterase (CbE) were drastically inhibited in the azamethiphos-treated group, 24 h after the dose. No inhibition was detected for BChE and CbE activities in the methomyl-treated group, 24 h after the dose. In the second treatment, the birds died or were sacrified 3 h after the dose. Serum BChE and brain acetylcholinesterase (AChE) were strongly inhibited after treatment with both insecticides. Serum CbE, hepatic microsomal CbE and 7-ethoxyresorufin dealkylation activities were also inhibited. A statistically significant correlation between serum BChE and brain AChE was found at lethal and sublethal doses of these xenobiotics. The experimental results indicate that the nondestructive biomarker BChE can give an early qualitative and semi-quantitative warning of the toxic effects of organophosphate and carbamate insecticides in birds.

Polycyclic aromatic hydrocarbons (PAHs) in subcutaneous biopsies of Mediterranean cetaceans

The aim of the present study was to measure polycyclic aromatic hydrocarbon (PAH) levels in free-ranging Mediterranean cetaceans as they are likely to cause chemical stress in the organisms of this basin. Blubber samples were collected from live specimens of fin whales (Balaenoptera physalus) and striped dolphins (Stenella coeruleoalba) by means of biopsies, a non-destructive biological method. Fin whales were sampled in the Ligurian Sea, whereas striped dolphins were collected in the Ligurian and the Ionian Seas. A fingerprint of 14 PAHs was obtained for both species. In whales, the median value of total PAHs was 1970 ppb fresh weight (f.w.) while median carcinogenic PAH values were 89.80 ppb f.w.; in dolphins, the median values of total and carcinogenic PAHs were 29,500 and 676.00 ppb f.w., respectively. The different PAH values between the two species can be attributed to the different positions they take in the Mediterranean food web. The sampling period significantly influenced PAH concentrations of fin whales.

Review: Porphyrins as Biomarkers for Hazard Assessment of Bird Populations: Destructive and Non-destructive Use

In this review the biochemical, metabolic and toxicological significance of porphyrins in birds is examined, and their use as biomarkers of exposure to xenobiotics and heavy metals is explored. Laboratory studies pinpointing the main classes of compounds that alter porphyrin profiles are described, as well as those which defined the resulting porphyrin profiles and target organs. Field studies in which the biomarker was validated on natural populations of several species of birds are then reviewed. We finally illustrate their potential as a non-destructive biomarker suitable for situations in which destructive sampling of natural populations must be avoided, suggesting the implementation of this approach.

Blood esterase inhibition in birds as an index of organophosphorus contamination: field and laboratory studies.

With the aim of developing a nondestructive biomarker (serum ‘B’ esterases) for monitoring bird populations exposed to azamethiphos S((6-chloro-2-oxooxazolo(4,5-b)pyridin-3(2H)-yl)methyl) 0,0-dimethyl phosphorothioate (9Cl), parallel laboratory and field studies were performed. In japanese quail (Coturnix coturnix japonica) treated with azamethiphos at 10 mg kg−1, serum butyrylcholinesterase (BChE) and carboxylesterase (CE) activities were inhibited by 88% and 35%, respectively, after 24 h. A gradual recovery was observed after 48 and 72 h. With a higher dose (50 mg kg−1), the birds died, or were killed 3 h after dosing. Serum BChE and brain acetylcholinesterase (AChE), in birds that were sampled at 3 h, were inhibited by 98% and 92%, respectively. Hepatic microsomal and serum CEs were also inhibited. There was a statistically significant correlation (r=0.9808, p<0.001) between the activities of serum BChE and brain AChE. Swallows (Hirundo rustica) nesting in a stable treated with azamethiphos showed a drastic reduction in BChE (56%) and CE (36%) 24 h after treatment of the stable.

Porphyrins in excreta: A nondestructive biomarker for the hazard assessment of birds contaminated with PCBs

The main aims of this paper are: a) to propose the use of porphyrins in excreta (nondestructive biomarker) as an alternative to the traditional liver porphyrins (destructive biomarker) in the hazard assessment of birds contaminated with PCBs and; b) to propose the use of a fast and inexpensive fluorimetric assay, proposed by Grandchamp et al. [1], as an alternative to HPLC for rapid evaluation of porphyrins in biological samples. The proposals were tested in the laboratory using three groups of Japanese quail (Coturnix coturnix japonica), with commercial bird food containing different concentrations of Arochlor 1260 for 25 days, and an untreated control group of the same birds.