Necrophilous insect succession on rabbit carrion in sand dune habitats in northern France.

Six freshly killed rabbit carcasses were exposed in different habitats in the coastal dune massif of Ambleteuse (northern France) during the spring seasons of 1996 and 1997. In total, 66 arthropod species were collected during the decomposition of these carcasses, and particular attention was paid to taxa of necrophilic significance. The pattern of insect activity was recorded and is discussed in relation to meteorological conditions. One significant feature in this study was the delay in initial oviposition by flies during 1996, which demonstrates the need for caution in estimation of postmortem interval by entomological techniques in early spring. Although daily temperatures may be favorable for adult fly activity, flies may be absent because of small population size and low nocturnal temperatures during this period.

Morphine perfused rabbits: a tool for experiments in forensic entomotoxicology.

In order to establish an animal model for entomotoxicological studies, the kinetics of morphine elimination from blood after a single intravenous injection of morphine and the concentration of morphine in tissues following a continuous perfusion were studied. The aim of these experiments was to obtain controlled morphine tissue concentrations similar to those encountered in fatal human heroin overdoses. These tissues can be used as a food source for developing fly larvae in entomotoxicological studies. In the single injection experiment, seven rabbits were administered 1 or 2 mg/kg body weight of morphine chlorhydrate via the main ear artery. Blood samples of 200 microL were removed regularly via a catheter. Morphine concentration was determined using RIA techniques. Morphine was found to be first rapidly distributed and then slowly eliminated, following a two-exponential equation. Elimination of morphine from blood can be described as a two-compartment model. Constants of the equation were determined using the Kaleidagraph program. Using those constants, the main pharmacokinetics parameters were calculated. Results of these parameters showed the following: clearance from 13.3 to 16.2 L.h.1, half-life of the distribution phase from 0.6 to 0.9 min, and half-life of the elimination phase from 21 to 26 min. These results were used to calculate the rate of perfusion of morphine for rabbits to obtain desired, controlled, and constant concentrations of morphine in tissues. In the second experiment, three rabbits received a perfusion of morphine intravascularly at a rate of 2 mg/kg/h for a period of 3 h. These rabbits were sacrificed and analyses performed on several abdominal and thoracic organs. Results showed that the concentrations of morphine differed according to the organ analyzed, but were reproducible for organs between animals. These concentrations were similar to those normally encountered in cases of human death due to heroin overdoses.

Determination of drug levels in larvae of Lucilia sericata (Diptera: Calliphoridae) reared on rabbit carcasses containing morphine.

This study concerns the determination of morphine concentrations in fly larvae reared on rabbits administered different concentrations of morphine and a correlation between concentrations of the drug in larvae and tissues. Three rabbits (R1, R2 and R3) were given dosages of 12.5, 25.0 and 50.0 mg/h of morphine over a 3 h period via continuous ear artery perfusion. These dosages and time of perfusion were calculated to create tissue concentrations of morphine similar to those encountered in human death due to overdose. Morphine blood level plateau was attained after 1 h of perfusion. A fourth rabbit was used as a control. To evaluate drug concentrations, tissues were sampled using a coelioscopic technique. Approximately 400 eggs of Lucilia sericata, all of the same age category, were placed in eyes, nostrils and mouth of each rabbit carcass. Larvae and puparia were regularly collected from each rabbit for toxicological analysis. The concentrations of the drug in the tissues sampled were determined to be similar to those normally encountered in human overdoses and were correlated with the dosage of morphine that had been administered. Morphine was detected in all larvae and pupae fed on tissues from carcasses administered morphine, except for puparia from the colony fed on the R1 animal which received 12.5 mg/h dosage of morphine. All samples from the control rabbit were negative for morphine. Concentrations of morphine in larvae reared on rabbit carcasses containing morphine were 30 to 100 times lower than the concentrations found in the tissues. A correlation between the tissue concentrations and larval concentrations was found in only 3rd instar larvae (80 to 140 h following hatching). No correlations were found between administered dosages, tissue concentrations and younger larvae, prepuparial larvae or puparia.

Effects of morphine in decomposing bodies on the development of Lucilia sericata (Diptera: Calliphoridae).

This study concerns the effects of morphine in tissues on the rate of development of Lucilia sericata (Diptera: Calliphoridae) using those tissues as a food source. Lucilia sericata is a species of fly commonly found on human corpses in Europe during the early stages of decomposition and thus of forensic interest. Three rabbits were administered 12.5, 25.0 and 50.0 mg/h of morphine chlorhydrate via ear perfusion over a period of 3 h. These dosages and duration of perfusion were calculated to give tissue concentrations of morphine similar to those encountered in fatal human overdoses. A fourth rabbit was used as a control. Following administration of the drug, rabbits were sacrificed and 400 eggs of Lucilia sericata, all of the same age, were placed in the eyes, nostrils and mouth of each rabbit. Developing larvae were sampled daily to determine growth rate and weight. Puparia and emerging adult flies were also sampled. Data were analyzed using analysis of variance (ANOVA) and Student's T-test. Results of this study show that an underestimation of the postmortem interval of 24 h is possible if the presence of morphine in tissues is not considered. This study demonstrates again the necessity of considering the possible effects of drugs in tissues on insect growth rates when estimating the postmortem interval using entomological techniques.

The oral health status of drug addicts. A prison survey in Lille, France.

The purpose of this study was to describe the prevalence of decayed, missing and filled teeth among the inmates of the jail of Loos (France) and to assess the impact of drug addiction, especially heroin, on these parameters of oral health. A representative sample was selected and two groups were compared: heroin addicts and non-drug addicts. The same dentist examined 93 inmates, males and females, from age 16 to 35. The results found a significant difference of DMFT index between the groups, with a higher value of DMFT for the heroin users. This investigation also highlighted atypical caries lesions among the heroin addicts.

Morphological analysis of the neurons in the area of the hypothalamic magnocellular dorsal nucleus of the guinea pig.

In the guinea-pig hypothalamus, a group of enkephalinergic cells forms a well-circumscribed nuclear area called the magnocellular dorsal nucleus (MDN). This nucleus gives rise to a prominent projection to the lateral septum: the hypothalamo-septal enkephalinergic pathway. In the present study, MDN neurons visualized by Golgi impregnation were subjected to morphological analysis in order to define the potential segregation of cellular types within the MDN. This study was complemented by additional observations of MDN neurons intracellularly injected by Lucifer yellow (LY) or horseradish peroxidase (HRP) during the in vitro incubation of hypothalamic slices. The following results were obtained from the analysis of 200 neurons: 163 Golgi-impregnated cells plus 37 injected cells (LY = 14; HRP = 23). Thirteen HRP-injected cells were precisely located in the MDN and 10 were located in the perifornical area surrounding the MDN. Four different cellular types were identified. Type-I neurons (41%) displayed a globular perikaryon, a variable number of primary dendrites that were poorly ramified, no preferential orientation, and an axon emerging from the perikaryon. Type-II neurons (30.5%) had a triangular perikaryon, three well-ramified primary dendrites, an orientation perpendicular to the third ventricle, and an axon emerging from the perikaryon. Type-III neurons (22%) exhibited a spindle-shaped perikaryon, two opposed well-ramified primary dendrites, an orientation perpendicular to the third ventricle, and an axon emerging from a primary dendrite. Type-IV neurons (6.5%), showed a globular perikaryon, a variable number of primary dendrites, poorly ramified dendrites, an orientation parallel to the third ventricle, and an axon whose orientation could not be identified. Neurons labeled after intracellular injection belonged to the first three cellular types.

Study of the efferent connections of the enkephalinergic magnocellular dorsal nucleus in the guinea-pig hypothalamus using lesions, retrograde tracing and immunohistochemistry: evidence for a projection to the lateral septum.

The efferents of enkephalin-immunoreactive neurons in the magnocellular dorsal nucleus of the guinea-pig were studied using different neuroanatomical methods and indirect immunocytochemical technique. Following unilateral implantation of the fluorescent dye 4',6-diamidino-2-phenylindole in the lateral septal nucleus, retrogradely-labeled perikarya were found in the magnocellular dorsal nucleus. These labeled perikarya reacted with antiserum against enkephalin, demonstrating that enkephalin-immunoreactive neurons in the magnocellular dorsal nucleus project to the lateral septal nucleus. In other experiments, complete bilateral lesions were produced in the magnocellular dorsal nucleus by electrocoagulation. Enkephalin-immunoreactive nerve fibers and terminals were totally depleted in the lateral septal nucleus. This confirms that septal enkephalin-immunoreactive terminals originate in the magnocellular dorsal nucleus and further suggests that this nucleus is the source of all the enkephalin-immunoreactive material found in the septum. Experiments utilizing two different fluorescent dyes, 4',6-diamidino-2-phenylindole and propidium iodide, injected in each side of the lateral septal nucleus, respectively, demonstrated that the magnocellular dorsal nucleus gives off axon collaterals to both sides of the septum, since double-labeling of individual cell bodies was detected in the nucleus. By relating this finding to the results obtained after unilateral destruction of the nucleus, which caused an incomplete loss of enkephalin- immunoreactive material in the lateral septal nucleus ipsilaterally, it is suggested that the enkephalinergic hypothalamo-septal pathway contains unbranching neurons projecting ipsilaterally and branching neurons distributing fibers ipsilaterally and contralaterally. Lesion experiments, and experiments based on the retrograde axonal transport of horseradish peroxidase after intravenous injections, demonstrated that the magnocellular dorsal nucleus contributes neither to the tubero-infundibular nor to the hypothalamo-neurohypophyseal tracts. The lateral septal nucleus receives numerous aminergic and peptidergic projections, indicating the potential importance of this region in physiological and behavioral events. In the guinea-pig, the well-demarcated enkephalinergic pathway demonstrated in this study provides a convenient model for the experimental study of the enkephalinergic innervation of the lateral septal nucleus.