Effects of bait age, larval chemical cues and nutrient depletion on colonization by forensically important calliphorid and sarcophagid flies

Species colonization patterns on corpses and the frequency of carrion fly oviposition and larviposition are affected by decomposition stage and previous maggot colonization. This study investigated these effects on meat bait colonization by Victorian Diptera of forensic importance. Bait treatments were: 'aged' (aged for 4 days at 22 °C, allowing some decomposition); 'nutrient-depleted' [aged for 4 days at 22 °C with feeding Calliphora vicina (Robineau-Desvoidy) (Diptera: Calliphoridae) larvae]; 'extract' (fresh bait mixed with liquid formed by feeding C. vicina larvae), and 'fresh' (untreated control bait). Statistical analysis (α = 0.05) revealed that colonization frequency differed significantly among treatments (Welch's F 3,18.83 = 4.66, P < 0.05). Post hoc tests showed that fresh and extract baits were colonized extensively throughout the experiment with no significant difference, whereas the colonization of nutrient-depleted baits was significantly lower. This suggests that larval digestive enzymes, larval excreta and cuticular hydrocarbons have less effect on colonizing Diptera than the nutritional content of meat. The colonization of aged baits did not differ significantly from that of fresh, extract or nutrient-depleted baits. A further experiment testing 'very aged' (aged for 8 days at 28 °C), 'larvae-added' (fresh bait with C. vicina larvae added before placement) and 'fresh' (untreated control) baits revealed that very aged baits were colonized significantly less frequently than either fresh or larvae-added baits (Welch's F 2, 6.17 = 17.40, P < 0.05).

Optical coherence tomography : age estimation of Calliphora vicina pupae in vivo?

Necrophagous blowfly pupae are valuable contributors to the estimation of post-mortem interval, should an accurate age estimate be obtained. At present, this is reliant on a combination of rearing and destructive methods conducted on preserved samples, including morphological observation and gene expression analyses. This study demonstrates the use of optical coherence tomography (OCT) as a tool for in vivo morphological observation and pupal age estimation. Using a Michelson OCT microscope, alive and preserved four and ten-day old Calliphora vicina pupae were scanned in different orientations. Two and three-dimensional images were created. Morphological characteristics such as the brain, mouthparts and legs were identifiable in both living and preserved samples, with distinct differences noted between the two ages. Absorption of light by the puparium results in a vertical resolution of 1-2 mm, preventing observation of deeper tissues. The use of contrast agents or a longer wavelength laser would improve the images obtainable. At present, the data suggests OCT provides a primary view of external and internal morphology, which can be used to distinguish younger and older pupae for further analysis of age and PMI estimation.

Comparação dos padrões de atratividade de Hermetia illucens (Diptera, Stratiomyidae) associada a carcaças de Rattus norvergicus enterradas e tratadas com hormônios esteróides

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Taxa de desenvolvimento de Chrysomya albiceps(Wiedemann) (Diptera: Calliphoridae) em dieta artificial acrescida de tecido animal para uso forense

Forensic entomology uses biological and ecological aspects of necrophagous insects to help in criminal investigations to estimate the post-mortem interval (PMI) or to determine the cause of death. Recent papers demonstrated that the presence of toxins in decomposing tissues may alter the insect developmental rate of insects exploiting such tissues as food. Thus, preliminary tests with artificial diets in laboratory are necessary to create a database to investigate and quantify the modifications that can occur with the collected insects from a criminal scene, avoiding any errors on the PMI estimates. The present study aimed to evaluate the developmental rate of Chrysomya albiceps (Wiedemann) reared on: a) artificial diets containing animal tissues: bovine liver (D1), raw muscle (D2), stomach (D3), and chicken heart (D4); b) artificial diet without animal tissue (D5); and c) a control group (C), which had only meat. The efficiency of each substrate was assessed by immature weight gain (mg), larval developmental time, larval and pupal survival, emergence interval and adult size. D1 to D4 diets did not restrict C. albiceps development; however, larvae reared on D1 and D2 diets presented a lower adult emergence rate. D3 and control group showed similarities regarding the efficiency parameters (rate and emergence interval). Thus, the use of diet D3, artificial diet with stomach, is the most recommended.

Occurrence of Hymenoptera on Sus scrofa carcasses during summer and winter seasons in southeastern Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Theoretical approaches to forensic entomology: II. Mathematical model of larval development

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Análise dos processos de decomposição e sucessão ecológica em carcaças de suínos (Sus scrofa L.) mortos por disparo de arma de fogo e overdose de cocaína e protocolo de procedimento diante de corpo de delito

Pós-graduação em Biologia Geral e Aplicada - IBB

Levantamento de sarcofagídeos(Diptera) do Brasil incluindo a caracterização molecular de Peckia (Pattonella) intermutans (Walker)

Pós-graduação em Biologia Geral e Aplicada - IBB

Análise faunística de dípteros necrófagos: ecologia e aplicação forense

Pós-graduação em Biologia Geral e Aplicada - IBB

Aspectos da biologia larval de Chrysomya megacephala (F.) (Diptera : calliphoridae): curva de crescimento e período de mais rápido desenvolvimento larval

Pós-graduação em Ciências Biológicas (Zoologia) - IBRC

Ethyl sulphate and ethyl glucuronide in vitreous humor as postmortem evidence marker for ethanol consumption prior to death

To clarify the circumstances of death, the degree of inebriation is of importance in many cases, but for several reasons the determination of the ethanol concentration in post-mortem samples can be challenging and the synopsis of ethanol and the direct consumption markers ethyl glucuronide (EtG) and ethyl sulphate (EtS) has proved to be useful. The use of a rather stable matrix like vitreous humor offers further advantages. The aim of this study was to determine the concentrations of ethanol and the biomarkers in the robust matrix of vitreous humor and to compare them with the respective levels in peripheral venous blood and urine. Samples of urine, blood from the femoral vein and vitreous humor were taken from 26 deceased with suspected ethanol consumption prior to death and analyzed for ethanol, EtS and EtG. In the urine samples creatinine was also determined. The personal data, the circumstances of death, the post-mortem interval and the information about ethanol consumption prior to death were recorded. EtG and EtS analysis in urine was performed by LC-ESI-MS/MS, creatinine concentration was determined using the Jaffé reaction and ethanol was detected by HS-GC-FID and by an ADH-based method. In general, the highest concentrations of the analytes were found in urine and showed statistical significance. The mean concentrations of EtG were 62.8mg/L (EtG100 206.5mg/L) in urine, 4.3mg/L in blood and 2.1mg/L in vitreous humor. EtS was found in the following mean concentrations: 54.6mg/L in urine (EtS100 123.1mg/L), 1.8mg/L in blood and 0.9mg/L in vitreous humor. Ethanol was detected in more vitreous humor samples (mean concentration 2.0g/kg) than in blood and urine (mean concentration 1.6g/kg and 2.1g/kg respectively). There was no correlation between the ethanol and the marker concentrations and no statistical conclusions could be drawn between the markers and matrices.

An integrated molecular biology and computational approach to CYP1A1 expression in human brain

The cytochromes P450 comprise a superfamily of heme-containing mono-oxygenases. These enzymes metabolize numerous xenobiotics, but also play a role in metabolism of endogenous compounds. The P450 1A1 enzyme generally metabolizes polycyclic aromatic hydrocarbons, and its expression can be induced by aryl hydrocarbon receptor (AhR) activation. CYP1A1 is an exception to the generality that the majority of CYPs demonstrate highest expression in liver; CYP1Al is present in numerous extrahepatic tissues, including brain. This P450 has been observed in two forms, wildtype (WT) and brain variant (BV), arising from alternatively spliced mRNA transcripts. The CYP1A1 BV mRNA presented an exon deletion and was detected in human brain but not liver tissue of the same individuals. ^ Quantitative PCR analyses were performed to determine CYP1A1 WT and BV transcript expression levels in normal, bipolar disorder or schizophrenic groups. In our samples, we show that CYP1A1 BV mRNA, when present, is found alongside the full-length form. Furthermore, we demonstrate a significant decrease in expression of CYP1A1 in patients with bipolar disorder or schizophrenia. The expression level was not influenced by post-mortem interval, tissue pH, age, tobacco use, or lifetime antipsychotic medication load. ^ There is no indication of increased brain CYP1A1 expression in normal smokers versus non-smokers in these samples. We observed slightly increased CYP1A1 expression only in bipolar and schizophrenic smokers versus non-smokers. This may be indicative of complex interactions between neuronal chemical environments and AhR-mediated CYP1A1 induction in brain. ^ Structural homology modeling demonstrated that P450 1A1 BV has several alterations to positions/orientations of substrate recognition site residues compared to the WT isoform. Automated substrate docking was employed to investigate the potential binding of neurological signaling molecules and neurotropic drugs, as well as to differentiate specificities of the two P450 1A1 isoforms. We consistently observed that the BV isoform produced energetically favorable substrate dockings in orientations not observed for the same substrate in the WT isoform. These results demonstrated that structural differences, namely an expanded substrate access channel and active site, confer greater capacity for unique compound docking positions suggesting a metabolic profile distinct from the wildtype form for these test compounds. ^

Value-adding Australian sardines: Factors affecting rates of deterioration in sardine (Sardinops sagax) quality during post-harvest handling

Deficiencies in sardine post-harvest handling methods were seen as major impediments to development of a value-adding sector supplying Australian bait and human consumption markets. Factors affecting sardine deterioration rates in the immediate post-harvest period were investigated and recommendations made for alternative handling procedures to optimise sardine quality. Net to factory sampling showed that post-mortem autolysis was probably caused by digestive enzyme activity contributing to the observed temporal increase in sardine Quality Index. Belly burst was not an issue. Sardine quality could be maintained by reducing tank loading, and rapid temperature reduction using dedicated, on-board value-adding tanks. Fish should be iced between the jetty and the processing factory, and transport bins chilled using an efficient cooling medium such as flow ice.

Modern diagnosis of Epstein-Barr virus infections and post-transplant lymphoproliferative disease

Infection by Epstein-Barr virus (EBV) occurs in approximately 95% of the world s population. EBV was the first human virus implicated in oncogenesis. Characteristic for EBV primary infection are detectable IgM and IgG antibodies against viral capsid antigen (VCA). During convalescence the VCA IgM disappears while the VCA IgG persists for life. Reactivations of EBV occur both among immunocompromised and immunocompetent individuals. In serological diagnosis, measurement of avidity of VCA IgG separates primary from secondary infections. However, in serodiagnosis of mononucleosis it is quite common to encounter, paradoxically, VCA IgM together with high-avidity VCA IgG, indicating past immunity. We determined the etiology of this phenomenon and found that, among patients with cytomegalovirus (CMV) primary infection a large proportion (23%) showed antibody profiles of EBV reactivation. In contrast, EBV primary infection did not appear to induce immunoreactivation of CMV. EBV-associated post-transplant lymphoproliferative disease (PTLD) is a life threatening complication of allogeneic stem cell or solid organ transplantation. PTLD may present with a diverse spectrum of clinical symptoms and signs. Due to rapidity of PTLD progression especially after stem cell transplantation, the diagnosis must be obtained quickly. Pending timely detection, the evolution of the fatal disease may be halted by reduction of immunosuppression. A promising new PTLD treatment (also in Finland) is based on anti-CD-20 monoclonal antibodies. Diagnosis of PTLD has been demanding because of immunosuppression, blood transfusions and the latent nature of the virus. We set up in 1999 to our knowledge first in Finland for any microbial pathogen a real-time quantitative PCR (qPCR) for detection of EBV DNA in blood serum/plasma. In addition, we set up an in situ hybridisation assay for EBV RNA in tissue sections. In collaboration with a group of haematologists at Helsinki University Central Hospital we retrospectively determined the incidence of PTLD among 257 allogenic stem cell transplantations (SCT) performed during 1994-1999. Post-mortem analysis revealed 18 cases of PTLD. From a subset of PTLD cases (12/18) and a series of corresponding controls (36), consecutive samples of serum were studied by the new EBV-qPCR. All the PTLD patients were positive for EBV-DNA with progressively rising copy numbers. In most PTLD patients EBV DNA became detectable within 70 days of SCT. Of note, the appearance of EBV DNA preceded the PTLD symptoms (fever, lymphadenopathy, atypical lymphocytes). Among the SCT controls, EBV DNA occurred only sporadically, and the EBV-DNA levels remained relatively low. We concluded that EBV qPCR is a highly sensitive (100%) and specific (96%) new diagnostic approach. We also looked for and found risk factors for the development of PTLD. Together with a liver transplantation group at the Transplantation and Liver Surgery Clinic we wanted to clarify how often and how severely do EBV infections occur after liver transplantation. We studied by the EBV qPCR 1284 plasma samples obtained from 105 adult liver transplant recipients. EBV DNA was detected in 14 patients (13%) during the first 12 months. The peak viral loads of 13 asymptomatic patients were relatively low (<6600/ml), and EBV DNA subsided quickly from circulation. Fatal PTLD was diagnosed in one patient. Finally, we wanted to determine the number and clinical significance of EBV infections of various types occurring among a large, retrospective, nonselected cohort of allogenic SCT recipients. We analysed by EBV qPCR 5479 serum samples of 406 SCT recipients obtained during 1988-1999. EBV DNA was seen in 57 (14%) patients, of whom 22 (5%) showed progressively rising and ultimately high levels of EBV DNA (median 54 million /ml). Among the SCT survivors, EBV DNA was transiently detectable in 19 (5%) asymptomatic patients. Thereby, low-level EBV-DNA positivity in serum occurs relatively often after SCT and may subside without specific treatment. However, high molecular copy numbers (>50 000) are diagnostic for life-threatening EBV infection. We furthermore developed a mathematical algorithm for the prediction of development of life-threatening EBV infection.