Entomotoxicology in Forensic Science
Entomotoxicologyin Forensic Science
Forensicscience is an essential discipline that deals with investigations onvarious manners and causes of death for appropriate legal actions tobe undertaken. It is a science that bridges the gap existing in themedicolegal system since it applies succinct investigative technologyto provide evidence (Cattaneo,2007).The discipline integrates the use of top notch skills and proficiencyto adequately cover the areas of postmortem studies and otherinvestigative tests aimed at finding the solution to different crimecauses and suspicious cases of death depending on various mechanismsof perpetuation (Iscan,& Steyn, 2013).
Applicationsof Different Components in Forensic Investigation
Theforensic scientists explore different areas to make their findingsprecise and well detailed. The methods used in the field are aimed atgauging appropriate causes and manners of death (Kobilinsky,2011).The study goes an extra mile to detect on the inbound logisticssurrounding a crime scene (Amendt,Campobasso, Gaudry, Reiter, LeBlanc, & Hall, 2007).In contingency to this, they scientists have developed variousmechanism to estimate the time of death so that discrepancies thatmight exist in the field are eliminated (Kobilinsky,2011).Among the parameters, the use of entomology is proving a provide aclear way of in-depth analysis of the time interval of death, causes,and the manner of death.
Thetrend is switching from other parameters that are equally used in thefield to more areas of entomology and deeper subsections (Amendt,Campobasso, Gaudry, Reiter, LeBlanc, & Hall, 2007).Through this, technological advancements developing based on theknowledge and extrapolative research on the forensic science. A newfield of study has been realized in the line of the survey.Entomotoxicology is a new field that has been accomplished in theexploration of the parameters, and it sets out to be a probing methodof determining the toxicology cases in forensic analysis (Cooper& Negrusz, 2013).The extrapolates on the how entomo- toxicology is used in used inforensic science and some of the major breakthroughs that have beenrealized in contingency to the new forensic analysis procedure(Amendt,Campobasso, Goff, & Grassberger, 2010).Additionally,the paper also explores on some of the deficiency areas that need tobe improved as far as entomo- toxicology is concerned (Amendt et al.,2010).
Overviewon the Use of Entomology in Forensic Investigation
Entomologyrefers to a detailed study of the insects from the perspective ofdifferent stages of the life cycle, metabolism, and theircharacteristics (Gennard,2012).In forensic science, entomology is vital in the estimation of thetime interval of death that is eventually followed up to ensure thata legal process is put in place (Gennard,2012).Application of the field is vital as it gives a clear and in-depthanalysis of the inbound logistics surrounding death cases ( Amendt etal., 2007). In many occasions, there are confounding factors thatcompromise effective deliberation of the forensic analysis (Amendt,Richards, Campobasso, Zehner, & Hall, 2011).The elements eventually end up as the likely possibilities of a falseestimation.
Theinsects mostly used in the forensic analysis are fruit flies andblowflies analysis (Amendt,Richards, Campobasso, Zehner, & Hall, 2011).Although many argue that, the main reason behind adoption of theinsects as the primary mechanism for estimation of the death timeinterval is genetic homozygosity with the humans, studies reveal thatthe adoption and use of the insects in forensic cases is based on theanalysis of different life cycle stages which lie in relation to theexact duration of the time of death (Amendt et al., 2011). Throughevaluation of different life cycles of the insects, forensicscientists can deduce the exact time of occurrence of deathretrospectively (Tsokos& Byard, 2012).
Forensicentomologists rely on the use of insect evidence to estimate the timeof death through having a keen analysis of the insects’ behavior,life cycle and the general metabolism (Eberhardt& Elliot, 2008).They evidence from the insects on, above, and below the body. A casestudy on the presence of blowfly in a corpse site indicates that theinsects are the first to arrive at a death scene within minutes(Eberhardt& Elliot, 2008).Evaluation of different stages of the life cycle indicates that theinsect has four developmental stages, and each stage is a marker ofthe specific interval that is vital in postmortem forensic analysis(Halide, Divrak, Taleb, 2016). In the case profile, it is indicatedthat blowfly eggs are first recovered in the death scene in less thaneight hours. Instar stages 1 and two are recovered within a span oftwenty hours and two and half days. Instar 3 stage is recovered afterfive days while the other subsequent stages lie pre-pupa, pupa, andadult lies within a span of eight and twenty four days (Halide,Divrak, Taleb, 2016). These stages are very essential since theyprovide the template for evidentiary material as well as permittingfurther forensic analysis (Byrd,& Castner, 2009).However, the disadvantage associated with insect evidence is that itjust estimates the close range time of death and the insects can beeasily affected by the fluctuation of the environment (Aak,Birkemoe, & Leinaas, 2011).The use of insect evidence is set to more implementations tosynergize more on getting succinct and precise results (Fiene, Sword,Vanlaerhoven, & Tarone, 2014). This makes it preferred theanalysis of the time of death alongside other methods like algormortis, rigor mortis, lividity, stages of body decomposition andevaluation of the gastrointestinal contents (Konigsberg,Herrmann, Wescott, & Kimmerle, 2008).
Whenconducting an investigation that applies the use of entomology, asystematic and well-outlined quality assured approach should beconsidered to allow adequate packaging and transportation ofentomological samples, ensuring that contamination is greatlyminimized as this will reduce the integrity of the collected evidence(Tsokos& Byard, 2012).The investigation is aimed at guaranteeing a chain of custody whichreinforces realizing of medico-legal jurisdictions. Great precautionis recognized in the investigation since it deals with the use ofliving organisms (Tsokos& Byard, 2012).
Forensictoxicology is mainly charged with conducting forensic analysis andinvestigation of adverse effects of drugs, poisons, and otherenvironmental chemicals in the body (Bumbrah,Krishan, Kanchan, Sharma, & Sodhi, 2012).The application of the practice involves a combination of differentdisciplines such as chemistry, pathology, and biochemistry toeffectively consolidate an evidentiary material surrounding a case ofthe subject (Bumbrahet al., 2012). Forensictoxicology professionals coordinate with pathologists, coroners, andmedical examiners in establishing a role of different drugs,chemicals, and poisons in the causation of death (Boghossian,Tambuscio & Sauvageau, 2010).Today, the area of forensics has changed to incorporate the use ofentomology and toxicology studies to give an in-depth analysis ofdifferent toxicology levels, which eventually aids in realizing morethoughtful ways of solving the crime cases suing expert post-mortemreport (Beyer,Drummer & Maurer, 2009).The combination of the two disciplines has given rise to the use ofentomo- toxicology in forensic investigation.
Investigationof Manner and Cause of death
Aforensic scientist is continually working around the clock to a reacha formidable way of estimating different causes and manner of death(Grant,Southall, Fowler, Mealey, Thomas, & Kinlock, 2007).The cause of death refers to underlying, primary or secondary factorsthat lead to cessation of life. On the other hand, the manner ofdeath relates to the mechanisms of which death occurs. Manners ofdeath according to forensic science, exists in different ways (Grant,Southall, Fowler, Mealey, Thomas, & Kinlock, 2007).The categories of the manner of death are accidents, natural deaths,suicides, homicides, and death due to misadventure. Forensicscientists are sometimes faced with a challenge of determining theexact cause and manner of deaths attributable to many confoundingfactors involved (Brunel,Fermanian, Durigon, & Grandmaison, 2010).As a result, the technology and the practice of forensic science areexperiencing a dynamic approach in which hands-on analysis of variousparameters surrounding the death cases are considered (Brunelet al., 2010).Integration of entomology and the knowledge on toxicology is provingto be a critical evaluative and probing measure that most of themedical researchers and forensic analysts are skewing towards.
Forensicinvestigation is changing an approach and skewing to the field ofentomo- toxicology, which acts a link between forensic entomology andtoxicology (Carvalho,2009).The emergence of the science-based investigative analysis mixedreactions based on science curiosity (Carvalho,2009).Many experts in the forensic science field continue to carry outdifferent practical projects to validate on the efficacy levels ofthe new method of crime detection and investigation of the postmortem interval of various death cases (Dayananda& Kiran, 2013).The subject refers to the in-depth analysis of toxins in insectsespecially the arthropods that feed on a corpse with the aim ofstudying toxicology based on the patterns of chemicals and theiradverse reactions (Dayananda& Kiran, 2013).The arthropods that are commonly used are flies and beetles. The deepforensic analysis is carried out using the insects to give clearforensic parameters over which legal jurisdiction can be enacted.Forensic entomology incorporates the use of insects in legal matterswhile toxicology involves investigation of the adverse effects oftoxins as well as connecting it retrospectively to the time of death(Dresen,Gergov, Politi, Halter, & Weinmann, 2009).Forensic entomotoxicology acts as an essential link between the twoinvestigative measures (Dresen,Gergov, Politi, Halter, & Weinmann, 2009).It offers a further probe based on experimental analysis of thechemical contents of the insects that are associated with places fcrime scene. This method also factors in the extraordinaryprecautions that are put in place to ensure that top notch qualityinvestigation is achieved. Contamination of evidentiary material isminimized at significant levels. Since the method involves the use ofinsects, care is taken to ensure proper collection of the insects,adequate and safety packaging, and appropriate transportation to theinvestigation sites (Dresen et al., 2009).
Forensicentomotoxicology narrows down on the study of insects as alternativetoxicological samples. The method is carried out systematically toensure that no necessary procedure is left out during theinvestigative experiment (Ojanperä, Kolmonen, & Pelander, 2012).It is mainly aimed at the precision and achieving reliability of therequired investigation (Carvalho,2009).Recent toxicology research dogma has advocated for the use of blood,urine, biopsy, and gastrointestinal contents for drug and poisondetection (Ojanperäet al., 2012).However, the need for more evidenced based research has prompted theadoption of forensic entomotoxicology, which sets in to provide asolution to use insects when the conventional collection materialsare not available for forensic analysis (Ojanpera et al., 2012).Insect evidence is commonly preferred by the forensic scientistsbecause they can investigate different parameters regarding thecause, and manner of death ((Bumbrah,Krishan, Kanchan, Sharma, & Sodhi, 2012).
Theprecision of estimates of etomotoxicology in many death casesinvolving drug intoxication has been a topic of discussion for mostof the past forensic conventions (Carvalho,2009).Many have argued in the line of adverse drug reactions realized inindividuals who have are subjects to poisoning (Carvalho,2009).Recent studies conducted reveal that the drug composition and thegeneral patterns in the bodies of the victims can be transferred tothe insects, which feed on the corpse bodies. The fly colonizationand ovipositional behavior of the insects are the key determinants ofthe development rates of carrion-frequenting insects that feed onsuch food sources (Ojanpera et al., 2012). The transfer confers thata study on the chemical and other gastrointestinal contents provide aretrospective analysis of the patterns of the poison in the humanbody.
Inregards to entomotoxicology, evaluation of the post mortem intervalis an integral step to establish good causal phenomena about theinvestigation of toxic compounds suing insects discovery (Villet,Richards & Midgley, 2009).Post-mortem interval indicates the time between the death and thecorpse discovery (Villet,Richards & Midgley, 2009).Among different methods used to gauge the post mortem intervals, thevalues attached the entomological data are considered most accurateand valuable ones. Insect values that are used to estimate the postmortem interval mainly rely on the insect activity (Brown,Thorne, & Harvey, 2012).The insect activity post-mortem interval refers to the period of fromthe insect arrival and colonization of the crime scene until thediscovery of corpse following death. Insect succession data isequally viable in analyzing different changes that occur within theforensic investigation period (Brown,Thorne, & Harvey, 2012).Calculation of the growth rate of the insects is also widely adoptedby most of the forensic investigators especially in cases where thedeath occurred in less than a month ago (Vass,2011).In this case, the variation of complexity in insects is analyzed. Anew approach used in the field is the microbial succession (Vass,2011).It looks at different rates in which the microbial organisms causebody decomposition. The stages of body decomposition are grouped in acascade depending on the complexity of the chemical changes realizedin the body systems (Vass,2011).The stages include fresh stage, bloat stage, initial skeletonization,late skeletonization, and dry decay stage (Verma,K., & Paul, 2013).Appropriate post mortem interval is thus gauged depending on variousstages.
Theknowledge of postmortem transfer is sufficient to minimize cases ofdiscrepancies in the evidence provided concerning a death scene crime(Paczkowski& Schütz, 2011).According to the surveys conducted on the incident cases of thepost-mortem transfer, it is realized that the process is gradual, andinvolves the establishment of the evidentiary material (NationalResearch Council, 2009).The process should be clearly understood to minimize chances ofconfounding factors obscuring evidence. Transfer of the body ismainly realized in instances of homicide where the perpetratortransfers the body to alter the original scene and change theevidentiary link. Forensic entomotoxicology takes this factor intoconsideration and therefore carries out investigations in differentcrime scenes detected so that a comparative data can be reached atand evaluated appropriately (Paczkowski& Schütz, 2011).A key consideration is taken to factor in information on the insectspecies distributions and preferred regions so that accurate data isgenerated by the different geographical areas (Sonet,Jordaens, Braet & Desmyter, 2012).The knowledge on the postmortem transfer is useful in linking up thechemical patterns in the insects and make an inferential conclusionto the identification of the cause and the manner of death of thecorpse identified (Sonetet al., 2012).
Historyof Forensic Entomotoxicology
Manysources are linking the emergence of forensic entomotoxicology toincreased use of entomological data in the forensic analysis of crimecases (NationalResearch Council, 2009).But a deeper review into the matter reveals that in the recent past,most deaths have been investigated to be drug related (NationalResearch Council, 2009).The greatest number of such death cases have been registered inAmerica. In many occasions, the deaths have been discovered after aperiod, which does not validate the tissues to be essential fortoxicological analysis (NationalResearch Council, 2009).Reversal to insect evidence has been widely adopted in many parts ofthe country (Murthy& Mohanty, 2010).The practice has since been widely adopted and more advancementmarked with evidence-based research forming the main emphasis. Theadoption of the entomotoxicology method started in 1980 whereentomologists began to detect drugs in insects with an aim that theprocess would turn out as a vital tool in the forensic investigation(Ojanperä,Kolmonen, & Pelander, 2012).Despite the gradual breakthroughs, most of the forensic scientistsargue that there is more research based evidence to back up theapplication of the method in forensic analysis ( Ojanpera et al.,2012).
Forensicentomology has been developed from an extensive history of researchdating back to the 13thcentury in China (NationalResearch Council, 2009).Studies validate that insects have been used for a long time in theinvestigative research for detection of crimes thus strengthening thepractice of forensic entomology (Zhou,Liu, Chang & Li, 2011).During these times, the correlation of different life cycle stages ofinsects was analyzed. Maggots correlation with the corpse wasinvestigated to determine the oviposition of the adult flies (Zhou,Liu, Chang & Li, 2011).However, there was not sufficient matching evidenced realized in thecase. Further probing into the study was needed.
Forensicentomology evidence has been widely used in many murder cases ofEurope during the 20thcentury with notable success (NationalResearch Council, 2009).This amassed greater interest in the second half of the century withmore interests shifted to the field of forensic entomology (Murthy& Mohanty, 2010).Employment of the scanning electron microscopy and DNA analysis hasimproved in the identification and even classification of the insectspecies (Wells& Stevens, 2008).Consequently, the overall performance index of forensic entomologyhas improved.
Advancementin the field of entomology has been fast approaching. The surge todevelop to more evidenced research necessitated emergence of forensicentomotoxicology (Murthy& Mohanty, 2010).This is after consultations and in-depth research that aimed atfinding out the crime deaths associated with drugs, and otherchemicals (Murthy& Mohanty, 2010).The possibility of Extraction of DNA tissue and a gunshot residuefrom the gut contents of feeding maggots made the use of insects inthe detection of the chemical substances and patterns of varioustoxins easier (Magni,Pazzi, Alladio, Vincenti, Brandimarte & Dadour,2016).
Theuse of entomotoxicology has since become a useful tool ininvestigating suicide, homicides or unattended deaths where toxinsare involved (Murthy& Mohanty, 2010).The procedure has conducted by placing the insect to some testsaiming at elucidating various active components that aid indetermining the relevance of a certain toxin to the death crimecases. The process of forensic entomology is a quite a complexprocess sine it involves systematic steps marked with qualitystandards to minimize contamination, and loss of evidence (Magni,Pazzi, Alladio, Vincenti, Brandimarte & Dadour,2016).
Stepsin Forensic Entomotoxicology
Thesampling of insects of interest is a crucial step in the method offorensic analysis (Konigsberg,Herrmann, Wescott, & Kimmerle, 2008).It detects the overall variability realized in the drug detection(Magni,Pazzi, Alladio, Vincenti, Brandimarte & Dadour,2016).For some of the toxicologists, insect sampling has become simple dueto the vast experience they have (Magni,Pazzi, Alladio, Vincenti, Brandimarte & Dadour,2016).However, the real sense of ensuring that the correct type of insectis collected requires particular ideology and equivalence to the task(Magniet al., 2016).During the sampling, caution should be taken to manage the insectwithout altering its morphology (DeCarvalho, 2009.Safe methods should be applied to maintain the integrity of theevidentiary links.
Asampling of insects occurs in the body, around, or at the recoverysite. Larvae and pupa stages especially of blowfly can be sampled atthis time (Magni,Pazzi, Alladio, Vincenti, Brandimarte & Dadour,2016).Many researchers reiterate that care should be taken to sample asmany of the insects as possible since some of the insects may comefrom different regions. Therefore, various sampling insects confer agreater variability of drugs. The insects should be collected aboutthe relevant sites. In insects, internal organs such as the livershould be sampled to be used in the analysis (Magni et al., 2016).The guidelines and standards for insect sampling should be adhered toenhance precision, quality, and the reliability of the results (Magniet al., 2016).
Fora sample to remain viable and fully valid for forensic entomo-toxicological analysis, conservation must be adhered to (França, Brandão, Sodré & Caldas, 2014). Once the specimens have beensampled from the body, they are subjecting to cleaning. In this case,deionizer is used, and the specimens frozen for storage at theappropriate temperature (França, Brandão, Sodré & Caldas,2014). They are kept in the condition until needed for analysis. Incases inorganic substances analysis, the insects are removed fromstorage and exposed more. Washing of the insects is carried outfollowed by proper drying to remove any external human fluid(omberlin,Mohr, Benbow, Tarone, & VanLaerhoven, 2011).The insects are then crushed and stored appropriately (França,Brandão, Sodré & Caldas, 2014). The preferred storage mediumis porcelain crucible at a constant temperature of 650 degreesCelsius for 24 hours span. The resultant mixture has a highcomposition of the metals required (Sukontason,Piangjai, Siriwattanarungsee, & Sukontason, 2008).The analysis of the metals is then carried out by using 70% nitricacid. This is mostly applicable in detecting heavy metals such ascases of lead poisoning in humans (Sukontasonet al., 2008).
Investigationsfor organics substances is started off by washing and drying thespecimens. A range of 1-10 grams of larvae is cut intensively(Konigsberg,Herrmann, Wescott, & Kimmerle, 2008).This removes any contaminant that will pose as a confounding factorthereby obscure observation (Malve,2016).An internal standard solution is then added. Homogenization of thespecimen then follows by use of 0.9% saline solution (Sukontason,Piangjai, Siriwattanarungsee, & Sukontason, 2008).The mixture is then centrifuged to get a supernatant. Strong acids orbases is then added to the resultant, and the mixture left overnightat a temperature of 65 degrees Celsius. The acids are removed, andthe organic acids are analyzed (Sukontason et al., 2008).
Investigationson the pharmacology postulate that pharmacokinetics vary in differentinsects. It entirely depends on the insect species, feeding activity,and the developmental stages (omberlin,Mohr, Benbow, Tarone, & VanLaerhoven, 2011).Apart from the analysis of organic and inorganic substances,bioaccumulation is also evaluated to detect the intensity of the druglevels (omberlin,Mohr, Benbow, Tarone, & VanLaerhoven, 2011).The study incorporates the use of necrophagous species belonging toColeoptera since they are the first to colonize the corpse, andtherefore offers a good presumptive evidence. They are thus definitein gauging appropriate post-mortem interval. Precautionary measure istaken during the procedure to ensure that no confounding factors arerecorded (omberlin,Mohr, Benbow, Tarone, & VanLaerhoven, 2011).It is realized that some products like cocaine, heroin, andmethamphetamine can induce effects in the insects used in theinvestigation such as the fruit flies (Konigsberg,Herrmann, Wescott & Kimmerle, 2008).Therefore, the concentration of the toxins is keenly followed todetect the actual changes and make an inferential result to theoriginal set condition of the investigation (omberlinet al., 2011).In cases of altered results realized, a thorough secondary probe willbe carried out to give to identify the exact concentration of thetoxins in the insects (Konigsberg.,2008)
TestsUsed to Determine the Toxic Substances Ingested by the Insects duringFeeding Stage
Theanalytical method focuses on identifying the trace elements of thetoxins consumed by the insects after passing a series of treatmentaimed at increasing the overall enumeration of some toxins (Maurer,2007).It is recognized for its gold standard adherence since it providesproficiency through allowing for 100% specificity in the test((Maurer,2007).The gas spectrometer utilizes capillary columns of differentdimensions regarding the film thickness, diameter, and the length(Bushby,Thomas, Priemel, Coulter, Rades, & Kieser, 2012).This allows for adequate evaluation of various properties of toxinsingested by the insects. Elucidation of the targeted toxins iscarried out after homogenization and addition of subsequent chemicalsthat expose the targeted toxins at a greater gradient. The moleculesor fragments of the toxins are then detected (Bushby,Thomas, Priemel, Coulter, Rades, & Kieser, 2012).The intensity of the same toxins is equally gauged. A correlation ismade based on some molecules identified within the chromatogram(Maurer,2007).Many forensic scientists are adopting this test method because it ishighly accurate since the readings are based on the gold standards,ensuring that high precision levels are attained (Bushbyet al., 2012).
Useof Liquid Chromatography-Mass Spectrometer
Themethod is mainly applied in forensic entomo- toxicology to detect thepresence of drugs, aldicarb, cocaine, and other metabolites in larvaefrom decomposed corpses (Maurer,2007).The metabolites that are detected in the larvae are sulfone andsulfoxide metabolites. The initial step involves solid –liquidextraction with the use of low temperature (Maurer,2007).Once the extraction is accomplished, the high-performance liquidchromatography is adjusted to maximize detection of toxins andmetabolites (Bushby,Thomas, Priemel, Coulter, Rades, & Kieser, 2012).The analytes are adjusted accordingly to allow proper detection andquantification of the larval products (Bushby,Thomas, Priemel, Coulter, Rades, & Kieser, 2012).Selectivity is achieved by adjusting and analyzing thechromatographic profiles of the toxins realized. An inferentialcomparison is finally made to correlate the level of toxins detectedto the amount digested by the insect (Bushbyet al., 2012).
LaserDiode Thermal Desorption-Tandem Mass Spectrometry
Thetechnique is based on the ionization (Meyer, Helfer, & Maurer,2014). Although most of the forensic laboratories still depend on themass spectrometry for good chromatographic assays, there is need toimplement the new technique, which detects trace evidentiary portionsof the toxins illumination (Meyer, Helfer, & Maurer, 2014). Theprocedure starts by dispensing a small amount of sample, mostly 2ulthe plates using a pipette (Wallace,2011).The solvent is given time to vaporize, leaving a mass of the toxin inthe chamber illumination (Meyer, Helfer, & Maurer, 2014). Theinfrared laser diode is used which is meant to embed with the toxicparticles, creating a greater illumination (Meyer, Helfer, &Maurer, 2014). The illumination generated is then analyzed and usedto quantify some toxins ingested by the insect (Meyer, Helfer, &Maurer, 2014).The technique is one of the major breakthroughs in thefield of forensic entomo- toxicology. There is a high level ofprecision realized by the method, and therefore validity andreliability are certain (Wallace,2011)
DNAExtraction from the Insects and the use of Polymerase Chain Reaction
Thetest works out in the coordination of two critical parameters. Itstarts from DNA extraction from empty puparium. (Primorac& Schanfield, 2014).In this method, empty puparium are obtained after the adults haveemerged at the advanced stages of body decomposition (Joshi& Deshpande, 2011).The extracted puparium is then processed in readiness to theextraction of DNA from its structure (Primorac& Schanfield, 2014).The adult flies are killed and soaked in 70% ethanol forpreservation. Initially, the collected puparium once obtained fromthe corpse are put in containers filled with sand and allowedsubjected to ambient of temperature of 24 degrees Celsius (Tarone, & Foran, 2011).A lid is used to cut off the air or any other sources ofcontamination that will lead to the loss of evidentiary material, andcompromise the overall intended result. The external surfaces of thepuparium are cleaned using a moistened swab (Primorac& Schanfield, 2014).A second swab is applied to remove the traces of water and any othersource of contamination. The cleaned partial cases are then dissectedusing a sterile blade, cut in tiny pieces, placed in 1.5ml Eppendorftubes, and processed immediately. DNA samples are extracted from thesamples using chelex-100 procedure, and PrepFlier applied biosystemprocedure (Tarone, & Foran, 2011).Once the DNA samples are ready, they are subjected to PolymeraseChain Reaction where an exponential expansion of the DNA fragment isachieved. This is the process by which the DNA fragments aremultiplied and increased in the overall count (Dunn,2014).
PCRuses an enzyme, Taq Polymerase, in which the subjected DNA samples isfirst denatured at a relatively high temperature, then annealed, andfinally exponentially expanded to get a greater number of detectedtoxins (Tarone, & Foran, 2011).The toxin derived from the DNA segments can then be quantified andinferred to some toxicity levels in both corpse and the insects(Joshi& Deshpande, 2011).Amplifications of the samples is done suing AmpF/STR NGMTM selectAmplification kit. The strength of different toxins are finallycompared and the peaks checked on the overall comparative study(Joshi& Deshpande, 2011).Gel electrophoresis is carried out to determine the exact quantity ofthe toxins, which is performed by comparison of the realized valuesto the baseline stated values (Dunn,2014).
EnzymeLinked Immunoabsorbent Assay ( ELISA)
Inthis method, the assay is carried with the main aim of identificationof the antigen and antibodies about the accumulated toxins in theinsects (Muuka,Hang’ombe, Nalubamba, Kabilika, Mwambazi, & Muma, 2011).Ideally, apart from enhancing the growth of some of the insects, thetoxins ingested also elicit a hypersensitivity reaction, which ismarked with the primary immunological response. The body of theinsects acclimatizes by producing antibodies to counter the action ofthe toxins which are viewed in this case as the antigens (Muuka,Hang’ombe, Nalubamba, Kabilika, Mwambazi, & Muma, 2011).Consequently, antigen-antibody reactions develop marked withdeposition of immunological complexes (Muukaet al., 2011).Forensic entomo- toxicologists take note of the phenomena and carryout tests alongside ELISA to detect the level of the immunocomplexesdeposition. One of the criteria used in this case is the complementsystem activation assays. The method identifies variable amounts ofcomplexes formed due to initiation of the complement system cascade.The overall result realized is then analyzed and extrapolated to sometoxins present in the gastrointestinal content of the insect`scomplexes (Tomberlin,Mohr, Benbow, Tarone, & VanLaerhoven, 2011).Thus,a conclusion is made that, the higher the immune complexes, thegreater the intensity of intoxication compound in the insect.
InELISA, an enzyme is used as a sandwich, which acts on the TMBsubstrate. Once the extracted samples are loaded into the wells(Tomberlinet al., 2011).Washing is done to remove any contamination, and the resultantmixture subjected to the antigen- antibody controls. The reaction isrealized in the process determined by the intensity of coloration (Muuka et al., 2011). Optical density is measured to gauge the exactquantity achieved.
Thetest is used to assess the level of toxins in the insects bydetermining the degree of inhibition of the enzyme. Cholinesterase isa vital enzyme in insects that coordinate the activity of the nervoussystem (Hofer,Eisenbach, Lukic, Schneider, Bode, Brueckmann, & Dalpke, 2008).The coordination ensures a well-organized framework on the insects’body physiological functions (Hofer,Eisenbach, Lukic, Schneider, Bode, Brueckmann, & Dalpke, 2008).It also acts as the transmitter substance, facilitating thetransmission of chemical signals associated with the coordination ofthe nervous system (Hofer,Eisenbach, Lukic, Schneider, Bode, Brueckmann, & Dalpke, 2008).Ideally, the presence of toxins in the insects is estimated to reducethe overall efficiency of the enzyme. The toxins act in anantagonistic fashion, barring useful functionality of the enzyme(Cooper& Negrusz, 2013).As a result, low enzyme activity is realized marked with anuncoordinated nervous response. Therefore, cholinesterase inhibitiontest is done by checking on the overall efficacy of the enzyme incontingency to the level of toxins Hofer,Eisenbach, Lukic, Schneider, Bode, Brueckmann, & Dalpke, 2008).An inference is then made based on the resultant enzyme affinity andperformance index Hofer,Eisenbach, Lukic, Schneider, Bode, Brueckmann, & Dalpke, 2008).
Thetest act as an alternative toxicological analysis method to detectsome drugs, chemicals, and other toxins in insects of forensicimportance (Dabbs,2013).The method marks unimportant histological procedure that preciselyidentifies the level of toxicity in insects. It requires a detailedhistological procedure (Dabbs,2013).The extracted substances from the insect`s must t are stained to makethem more distinct and for easier analysis. Different fixatives areused to make the stains maintaining the firm structure and to generalpreserve the specimens (Cooper& Negrusz, 2013).Fixatives of preference for the method are 4% phosphate-bufferedparaformaldehyde, Kahle’s solution ( KS), ethanol in differentconcentrations, Carnoy’s Fluid, and ethanol associated with PP andCF but in different concentrations (Dabbs,2013).Before the start of the staining exercise, all the reagents areassembled in place to allow the systematic flow of the procedure.
Timeof fixation is considered an important factor since it determines theoverall integrity of the staining procedure. The dipteran larvae’stissue once isolated is subjected to fixation for a given periodbefore subjection to various stages of staining (Dabbs,2013).The sample must be collected and be prepared by strictly adhering tothe sample preparation protocol (Dabbs,2013).All the standard guidelines must be followed to minimize chances ofcontamination that will reduce the overall quality of the expectedresult (McIntyre,2014).The Immunochemical Test aims at detecting the presence of immunecomplexes that are deduced by conjugating antibodies (McIntyre,2014).The conjugated antibodies introduced to the system serves to bindwith the toxins, narcotoxins, drugs, and chemicals viewed as theantigen. The level of binding is thus determined to give a goodinferential result on some toxins ingested by the insect. Differentantibodies are used for conjugation depending on various toxins beingtested (McIntyre,2014).This explains that the binding affinity of the conjugated antibodiesvaries accordingly with different toxins (McIntyre,2014).Thus, binding efficacy is enhanced through conjugation of monoclonalantibodies that show coherence to the expected toxins. For example,in the immunochemical test for detection of cocaine using amonoclonal benzoylecgonine antibody from mouse, an introduction ofPeroxidase-conjugated anti-mouse Immunoglobulin and visualized by3,3- diaminobenzidine method, the method proved a useful histologicalmethod that did not compromise antigenicity (McIntyre,2014).
MajorConcepts in Entomotoxicology
TemperatureQuantification of Maggot Masses
Temperatureis a major contributor in the determination of postmortem intervals(Matuszewski & Szafałowicz, 2013). The variation in temperatureranges indicates different time frames that the corpse has takensince death (Gennard,2012).About the field of entomotoxicology, temperature changes should beconsidered in the insects to gauge a proper correlation of theinitial ambient temperature of the corpse to that in insects(Matuszewski & Szafałowicz, 2013). The temperature ranges aremajorly compared using blowflies since they give a variation forcomparison (Halide et al., 2016)
Activityand distribution of toxins in insects are affected by temperature(Matuszewski & Szafałowicz, 2013). Therefore, analysis of theparameters in temperature ranges acts as a key determinant of theoriginal processes of biotransformation, and other fundamentalprocesses responsible for altered amounts of toxins in the insects(Symes,Rainwater, Chapman, Gipson & Piper, 2008).Most of the forensic entomo- toxicologists have considered thisfactor, and are incorporating mechanisms to evaluate the temperaturechanges in contingency to the toxicity levels (Symes,Rainwater, Chapman, Gipson & Piper, 2008).
Inthe latest experiment, maggot masses of numerous Calliphoridaespecies have been used. It has been realized that most of the maggoteggs of the stated species form aggregations of larvae duringdevelopmental stage particularly in the feeding stage (Symes,Rainwater, Chapman, Gipson & Piper, 2008).The changes in complexity lead to a possible increase in temperature.An experiment was conducted to investigate the interrelation betweenthe maggot numbers in totality and generation of heat (Aak,Birkemoe & Leinaas, 2011).To accomplish the experiment single species aggregates of Lucillasericata subjectedto a favorable environment in the laboratory to allow them to growand develop at a normal temperature of 22 degrees Celsius (Symes,Rainwater, Chapman, Gipson & Piper, 2008).A follow up on growth wa made at different intervals, and theinternal temperature measured intermittently to gauge on the changesdue to metabolism. Results indicated that 1200 mass size elicitedtemperatures that were slightly higher initial ambient temperature(Symes et al., 2008). The result points out that the microclimate ofmass can vary notably from the original set conditions, whichinfluences the larval development rates, and therefore this measureis efficient and equally valid. Determination of the postmorteminterval and the toxicity levels should, therefore, incorporate thestandard so that accuracy is increased (Symes et al., 2008). Forensicscientist applies the use of specific indicators to gauge the levelof toxicity and to assess the post mortem interval, which is inferredto be the time that has elapsed from the death of individual untilinvestigations concerning the death is commenced (Kobilinsky,2011).
Pre-appearanceIntervals in Carrion Insects
Determinationof postmortem interval and appropriate pre-appearance intervals areareas that attract most attention of the forensic entomologists andforensic entomo- toxicologists (Matuszewski,2011).Pre-appearance interval is in many cases misunderstood due toconfounding factors realized in its determination (Matuszewski,2011).Most of the forensic scientists argue that it is a presumptive timeto most of the highly carried out detective mechanisms to estimatethe postmortem intervals (Tomberlin,Mohr, Benbow, Tarone & VanLaerhoven, 2011).However, this perception has been viewed differently according tomany cases of advanced researchers validating on the issue.
Pre-appearanceInterval refers to the time limit that precedes the appearance of anarthropod insect on a cadaver (Wallace,2011).Its length is congruent to the temperature changes. In some carrioninsects such as beetles, a variation of temperature affects theiroverall distribution in the area of the cadaver (Wallace,2011).Other factors about the temperature changes are equally vital inexplaining the variation in the pre-appearance interval. Such factorsinclude repellants being present in the body, physical barriers,which limit the dispersion of attractants (Tomberlinet al., 2011).Amongst these parameters, temperatures are considered the mostsignificant factor to consider in eliminating discrepancies thatmight be incurred in the preliminary stages in the forensic analysis(Wallace,2011).The field has attracted curiosity from most of the forensicentomologists who have developed models to investigate the trends inthe temperature variation and how it affects the pre-appearanceintervals. Given this, two models of different carrion insects havebeen developed (Nagy,2010).The first model uses the larval stage while the second modelincorporates the adult stage (Wallace,2011).The two models are then analyzed comparatively to determine theoverall efficacy of the experiment, and to enhance precision and topnotch evidence (Tsokos& Byard, 2012).It is realized that temperature variation apart from the estimationof the time of death, it determines both pre-appearance intervals andthe postmortem intervals (Nagy,2010).The experiments however much proved to be vital, validation of thefindings has not fully been established as most of the forensicspecialists still feel that more research should be done to determinethe coherence of the variables by investigation the inbound logisticthat relates pre-appearance interval, toxicity levels, and postmorteminterval (Wallace,2011).
Forensicentomo- toxicologists have identified a gap in this area, and areworking on developing research that aims at determining the variationof the toxicity levels in the carrion insects in reaction to thefactors surrounding the pre-appearance interval (Wallace,2011).
.The presumptive study is aimed at finding out a validation mechanismon the changes in the strength of toxins ingested by the insect byconducting a retrospective linking analysis to some of the factorsespecially the temperature that could mark the pre-appearanceInterval (Wallace,2011).Other factors of external seasonal variation are also put intoconsideration. The pre-appearance interval therefore in differentinsects (Tsokos& Byard, 2012).The patterns are contingent to the cycles of the insects, which actas the presumptive determinants.
Useof Biomarkers in Carrion Insects’ Strategy
Mostof the forensic entomologists and forensic entomo- toxicologistsargue that the whole process of insect collection, preparation,preservation, and analysis tend to be hectic (Wells& Stevens, 2008).As a result, propositions are being made to induce specificallydesignated biomarkers in the carrion insects that will act as thepresumptive analysis tool (Wells& Stevens, 2008).This implies that insects’ system should be induced with speciallyformulated biomarkers that integrate the toxic components in insectsso that the toxins can be timely detected using Gas-LiquidSpectrometry or any other suitable method (Peters,2011).
Thesurveys that have probed into the new method, however, fear for theinstances of losing accuracy. They argue that most of the biomarkersmight be affected by the general metabolic functions of the insects(Tsokos& Byard, 2012).Again, some postulate that since the biomarkers will be embedded intothe deep body organs of the insects, there are likely chances thatthe binding affinity will be significantly reduced due to minimizingexposure sites (Wallace,2011).Such an effect will equally reduce the efficacy of toxins detection.
Theuse of biomarkers in insects though is a lucrative adventuremeasures should be taken to carry out evidenced based clinical trialswhere the affinity of the drugs, toxins, narcotoxins, and otherchemicals to the biomarkers be tested (Wells& Stevens, 2008). Genetic modification should also do on the biomarkers and orientthem appropriately so that genetic coherence is realized (Wells& Stevens, 2008).A presumptive analysis of the genetic coding reveals the correlationof the genetic predisposition to the toxicity levels (Zhou,Liu, Chang & Li, 2011).
ToxicityEffects in Insects
Thefact that forensic entomotoxicology focuses only on analyzing thechemical compounds, and toxins in cadavers using the carrion insectsdoes not fully cover the required parameters (Gosselin,Di Fazio, Wille, Fernandez, Samyn, Bourel, & Rasmont, 2011).There is a, however, an agitation that a comparative study should bemade evaluating the toxicity effects of the same compounds in theinsects to gauge the variation (Gosselinet al., 2011).Although some earlier interventions that has led to the emergence ofvarious insecticides have been made, no precise thin lineinvestigates concisely on the effects of the toxins in the insects(Gosselinet al., 2011). Thus toxicology bridges the gap of identification ofthe toxicity levels, and the resulting morphological changes itexerts in the bodies of the insects (Amendt et al., 2010)
Entomologicalspecimens are viewed to make excellent qualitative toxicologicalsamples (Malve,2016).However, a gap has been identified in the failure of conductingresearch based on developing an assessment to quantify theconcentration of different drugs, toxins, and chemicals using theentomological evidence (George,Archer, Green, Conlan, & Toop, 2009).There is no explicit knowledge of the drug patterns and standardsespecially about the adverse effects that is the central point ofinvestigation when determining the concentration of toxin levels inthe cadavers to infer an appropriate cause and manner of death(Flanagan,Taylor, Watson & Whelpton, 2008).It is realized that drug that can only be detected in larvae when therate of absorption becomes far much higher than the rate ofelimination. Also, the problem is also linked to the fact thatsamples of pupae and third instars larvae do not containconcentrations of the drugs (Sisco,Forbes, Staymates & Gillen, 2016).The observations are possible suggestions that drugs do notbio-accumulate in the whole life cycle of larvae.
Thereare also gaps in explaining the exact mechanisms in which the toxinsand other drugs get eliminated from the structure of their system(Malve,2016).It is logical enough to both analyze the adverse effects of toxicitylevels in the insects, their acclimatization immunologicalorientation, and toxins elimination (George,Archer, Green, Conlan, & Toop, 2009).The fact that no clear evidence-based research method has beenpostulated relating the three major concepts in forensic entomo-toxicology, has left most of the forensic experts theorizing thatelimination of the toxins from the system of larvae occasionallyoccurs if no constant supply of the toxin is in place (Wielgomas,Czarnowski & Jansen, 2012).The gaps evidenced in this field of entomology validate essence ofinfertility. Therefore, a lot of active research is needed in thearea (Wallace,2011).Once comparable results are found, the forensic entomo- toxicologistswill move a notch higher in advancing their investigations on thetoxicity effect and the postmortem intervals (Wielgomas,Czarnowski & Jansen, 2012).Consequently, a more advanced measure is set to be reached at thatdetermines the definite cause and manner of deaths in mostdrug-related deaths manifested many parts of the globe (Wielgomas,Czarnowski & Jansen, 2012).
MultipleColonization of a Cadaver by Insects
Multiplecolonization refers to a process where insects of different speciesfeed on the same cadaver (Picard& Wells, 2009).It has been realized in many places, a recent example found withinthe municipality of Jaboatao dos Guararapes in Brazil where a bodywas colonized by six species belonging to Diptera (Picard& Wells, 2009).According to forensic validation, the most abundant species wereChrysomyabicepsand Chrysomyamegacephaly. Othercases of cadaver colonization have also been reported from differentparts of the globe (Picard& Wells, 2009).Multiple cadaver colonization provides forensic entomologists withsevere in estimating accurate time of death (Oliveira,& Vasconcelos, 2010).This is attributed to different life cycle stages of the variousinsects. It is an oblivious analysis if a scientist narrows down toonly one insect and investigate because of all the variables, in thiscase, are important, and must be examined (Oliveira,& Vasconcelos, 2010).
Manyconfounding results are realized in the cases of multipleorganizations. The scientist, in this instance, must first evaluateall the inbound phenomena, highlight all the species, and configureout their adverse effects, sources of attractants, feeding activity,and the general life cycle sequence (Oliveira,& Vasconcelos, 2010).Multiple colonization of cadavers by insects provides a gap for theforensic entomo- toxicologists (Pohjoismäki,Karhunen, Goebeler, Saukko& Sääksjärvi, 2010).They are tasked to find out various toxicity levels of the insectspecies and evaluate the overall result on a comparative basis sothat a certain toxin can be realized entomo- toxicologists(Pohjoismäki,Karhunen, Goebeler, Saukko& Sääksjärvi, 2010).However, one challenge is attached to the intervention (Vasconcelos,Soares & Costa, 2013). Different toxins react differently todifferent insect species. This, in turn, affects their mode of growththerefore intermittent cases can be realized (Vasconcelos, Soares &Costa, 2013).
Guidelinesand Law Requirements in Forensic Entomology and Entomotoxicology
Insectevidence is a vital process used to gauge the death time frame(Dadour& Harvey, 2008). Likewise, determination of the cause and manner of death has beenmade easier by the integrative analysis of the insects’characteristics, life cycle, feeding activity, and toxicologicaleffects (Malve,2016).In some cases, the reliability of forensic entomology has beencompromised due to the unrealistic experimental settings, which doesnot allow immature collection of insects of different species onhuman bodies at the death (Dadour& Harvey, 2008).Therefore, jurisdiction standards and policies have been put in placeto regulate the overall practice to ensure that it remains within therequired legal standards (Dadour& Harvey, 2008).
Legal,ethical restraints and bureaucratic lowers cadavers usage forforensic studies in some countries like Brazil. This allows access tothe homicide victims to be only limited to police personnel(Mozayani,& Noziglia, 2010).It is evident that the availability of human bodies is quite erraticand therefore medico-legal practices conducted on it usually lead toloss of evidence (Amendtet al., 2010).This has been a critical issue where most of the forensic officershave found themselves at crossroads with most of the ethical reviewgroups (Mozayani,& Noziglia, 2010).The officers take their time sample and evaluate insects found at thecrime scene whether it is multi colonization or single colonization(Singh,& Sharma, 2008).The ethical considerations, however, considers that processes aimedat restoring the dignity of the corpse should be immediatelyconducted (Malve,2016).The bone of content lies in the fact that, there is a significantloss of entomological evidence if such activities are directlyconducted (Mozayani,& Noziglia, 2010).
Entomologicalevidence use in the investigation of the crime cases requires theforensic scientist to conduct a thorough collection of detailsinformation on the conditions of death cases such as abiotic factors,and access the cadaver (Skopp,2010).However, the processes should be carried out in strict adherence tothe stated legislation standards of the forensic science code ofethics and the general fundamental human rights (Mozayani,& Noziglia, 2010).The Forensic entomologists and cadres in entomotoxicology arerequired to carry out their investigations about the ethical policies(Skopp,2010).
Obtainingreal case reports and databases for further research inentomotoxicology is involving (Amendtet al., 2010).This is due to the difficulty in establishing advanced analyticaltechniques in the field (Skopp,2010).The process is expensive and time-consuming as it involves in-depthanalysis of the insects to relate a full result on the toxicologicalconcentration levels for the scientist to finally infer results basedon the actual experiment carried out (Skopp,2010).Another challenge in the field is skewed to the fact that postmortemdrug distribution and drug stability in humans and insects isundetermined, which makes various interpretations involving (Zhou,Liu Chang& Li, 2011).Additionally, use of human tissues like liver to rear larvaeexperimentally is considered unethical, and therefore Vitro researchon entomotoxicology has been difficult (Zhou,Liu Chang& Li, 2011).
Inconclusion, forensic entomotoxicology is a vital research field thatrequires extensive knowledge to thoroughly investigate on thepostmortem interval and come up with conclusive results, detailing onthe toxicity levels of the corpse through inferential analysis of thelevels using insects (Zhou,Liu Chang& Li, 2011).The method manifests advancement if forensic entomology, which idealinvestigates the death cases using a straight insect evidence thatevaluates the life cycle and the behavior characteristics of theinsects and deduce a result (Singh,& Sharma, 2008).Forensic entomotoxicologist differs from the latter since it probesdeeper to get a fact-based evidence on the drug-related cases (Singh,& Sharma, 2008).It is a field that is still open to wider innovations and top-notchcreativity enhancement focusing on the main topics that still raisedoubts such as bioaccumulation of toxins and insect metabolism ofdrugs.
Aak,A., Birkemoe, T., & Leinaas, H. P. (2011). Phenology and lifehistory of the blowfly Calliphora vicina in stockfish productionareas. Entomologiaexperimental et applicator, 139(1),35-46.
Amendt,J., Campobasso, C. P., Gaudry, E., Reiter, C., Leblanc, H. N., &Hall, M. J. (2006). Best practice in forensic entomology—standardsand guidelines. International Journal of Legal Medicine, 121(2),90-104. doi:10.1007/s00414-006-0086-
Amendt,J., Campobasso, C. P., Gaudry, E., Reiter, C., LeBlanc, H. N., &Hall, M. J. (2007). Best practice in forensic entomology—standardsand guidelines. Internationaljournal of legal medicine, 121(2),90-104.
Amendt,J., Campobasso, C. P., Goff, M. L., & Grassberger, M. (Eds.).(2010). Currentconcepts in forensic entomology (p.376). Dordrecht:: Springer.
Amendt,J., Richards, C. S., Campobasso, C. P., Zehner, R., & Hall, M. J.(2011). Forensic entomology: applications and limitations. Forensicscience, medicine, and pathology, 7(4),379-392.
Baird,D. J., Pascoe, T. J., Zhou, X., & Hajibabaei, M. (2011). Buildingfreshwater macroinvertebrate DNA-barcode libraries from referencecollection material: formalin preservation vs. specimen age. Journalof the North American Benthological Society, 30(1),125-130.
Bartol,C. R., & Bartol, A. M. (2011). Introductionto forensic psychology: Research and application.Sage.
Beyer,J., Drummer, O. H., & Maurer, H. H. (2009). Analysis of toxicalkaloids in body samples. Forensicscience international, 185(1),1-9.
Boghossian,E., Tambuscio, S., & Sauvageau, A. (2010). NonchemicalSuffocation Deaths in Forensic Setting: A 6‐YearRetrospective Study of Environmental Suffocation, Smothering,Choking, and Traumatic/Positional Asphyxia. Journalof forensic sciences, 55(3),646-651.
Brown,K., Thorne, A., & Harvey, M. (2012). Preservation of Calliphoravicina (Diptera: Calliphoridae) pupae for use in post-mortem intervalestimation. Forensicscience international, 223(1),176-183.
Brunel,C., Fermanian, C., Durigon, M., & de la Grandmaison, G. L.(2010). Homicidal and suicidal sharp force fatalities: autopsyparameters about the manner of death. Forensicscience international, 198(1),150-154.
Bumbrah,G. S., Krishan, K., Kanchan, T., Sharma, M., & Sodhi, G. S.(2012). Phosphide poisoning: a review of the literature. Forensicscience international, 214(1),1-6.
Bushby,S. K., Thomas, N., Priemel, P. A., Coulter, C. V., Rades, T., &Kieser, J. A. (2012). Determination of methylphenidate in Calliphoridlarvae by liquid–liquid extraction and liquid chromatography massspectrometry–Forensic entomotoxicology using an in vivo rat brainmodel. Journalof pharmaceutical and biomedical analysis, 70,456-461.
Byrd,J. H., & Castner, J. L. (Eds.). (2009). Forensicentomology: the utility of arthropods in legal investigations.CRC Press.
Cattaneo,C. (2007). Forensic anthropology: developments of a classicaldiscipline in the new millennium. ForensicScience International, 165(2),185-193.
Cooper,G., & Negrusz, A. (Eds.). (2013). Clarke`sanalytical forensic toxicology.Pharmaceutical Press.
Dabbs,D. J. (2013). Diagnosticimmunohistochemistry.Elsevier Health Sciences.
Dadour,I. R., & Harvey, M. L. (2008). The use of insects and associatedarthropods in legal cases: a historical and practicalperspective. ForensicApproaches to Death, Disaster, and Abuse,225.
Dayananda,R., & Kiran, J. (2013). Entomotoxicology. InternationalJournal of Medical Toxicology and Forensic Medicine, 3(2(Spring)), 71-74.
DeCarvalho, L. M. L. (2009). Toxicology and Forensic entomology.In Currentconcepts in forensic entomology (pp.163-178). Springer Netherlands.
Dresen,S., Gergov, M., Politi, L., Halter, C., & Weinmann, W. (2009).ESI-MS/MS library of 1,253 compounds for application in forensic andclinical toxicology. Analyticaland bioanalytical chemistry, 395(8),2521.
Dunn,M. J. (Ed.). (2014). Gelelectrophoresis of proteins.Elsevier.
Eberhardt,T. L., & Elliot, D. A. (2008). A preliminary investigation ofinsect colonization and succession on remains in NewZealand. ForensicScience International, 176(2),217-223.
Espeland,M., Irestedt, M., Johanson, K. A., Åkerlund, M., Bergh, J. E., &Källersjö, M. (2010). Dichlorvos exposure impedes extraction andamplification of DNA from insects in museum collections. Frontiersin Zoology, 7(1),2.
Fiene,J. G., Sword, G. A., Vanlaerhoven, S. L., & Tarone, A. M. (2014).The Role of Spatial Aggregation in Forensic Entomology: Table1. Journal of Medical Entomology, 51(1), 1-9.doi:10.1603/me13050
Flanagan,R. J., Taylor, A. A., Watson, I. D., & Whelpton, R.(2008). Fundamentalsof analytical toxicology.John Wiley & Sons.
França,J. D., Brandão, M., Sodré, F. F., & Caldas, E. D. (2014).Simultaneous determination of prescription drugs, cocaine, aldicarb,and metabolites in larvae from decomposed corpses by LC–MS–MSafter solid–liquid extraction with low-temperature partitioning.Forensic Toxicology, 33(1), 93-103. doi:10.1007/s11419-014-0255-4
Gennard,D. (2012). Forensicentomology: an introduction.John Wiley & Sons.
George,K. A., Archer, M. S., Green, L. M., Conlan, X. A., & Toop, T.(2009). Effect of morphine on the growth rate of Calliphora stygia(Fabricius)(Diptera: Calliphoridae) and possible implications forforensic entomology. Forensicscience international, 193(1),21-25.
Gosselin,M., Di Fazio, V., Wille, S. M., Fernandez, M. D. M. R., Samyn, N.,Bourel, B., & Rasmont, P. (2011). Methadone determination inpuparia and its effect on the development of Lucilia sericata(Diptera, Calliphoridae). Forensicscience international, 209(1),154-159.
Gosselin,M., Wille, S. M., Fernandez, M. D. M. R., Di Fazio, V., Samyn, N., DeBoeck, G., & Bourel, B. (2011). Entomotoxicology, experimentalset-up, and interpretation for forensic toxicologists. Forensicscience international, 208(1),1-9.
Grant,J. R., Southall, P. E., Fowler, D. R., Mealey, J., Thomas, E. J., &Kinlock, T. W. (2007). Death in custody: a historicalanalysis. Journalof forensic sciences, 52(5),1177-1181.
Halide.Nihal. Duygu, Divrak. Meriem, Taleb. (2016). Identification of aforensically and medically important blowfly: A case study of adultCalliphora vicina (Rob-Desvoidy) in Turkey identification of adultCalliphora vicina. European Journal of Forensic Sciences.Retrieved April 4, 2017.
Hofer,S., Eisenbach, C., Lukic, I. K., Schneider, L., Bode, K., Brueckmann,M., & Dalpke, A. H. (2008). Pharmacologic cholinesteraseinhibition improves survival in experimental sepsis. Criticalcare medicine, 36(2),404-408.
Iscan,M. Y., & Steyn, M. (2013). Thehuman skeleton in forensic medicine.Charles C Thomas Publisher.
Joshi,M., & Deshpande, J. D. (2011). Polymerase chain reaction:methods, principles, and application. InternationalJournal of Biomedical Research, 2(1),81-97.
Kobilinsky,L. (Ed.). (2011). Forensicchemistry handbook.John Wiley & Sons.
Konigsberg,L. W., Herrmann, N. P., Wescott, D. J., & Kimmerle, E. H. (2008).Estimation and evidence in forensic anthropology:age‐at‐death. Journalof forensic sciences, 53(3),541-557.
Lundgren,J. G., Ellsbury, M. E., & Prischmann, D. A. (2009). Analysis ofthe predator community of a subterranean herbivorous insect based onpolymerase chain reaction. EcologicalApplications, 19(8),2157-2166.
Magni,P., Pazzi, M., Alladio, E., Vincenti, M., Brandimarte, M., &Dadour, I. (2016). Blowflies & nicotine: an entomotoxicologystudy.
Malve,H. O. (2016). Forensic Pharmacology: An important and evolvingsubspecialty needs recognition in India. Journalof pharmacy & bioallied sciences, 8(2),92.
Matuszewski,S. (2011). Estimating the pre-appearance interval from thetemperature in Necrodes littoralis L.(Coleoptera:Silphidae). ForensicScience International, 212(1),180-188.
Matuszewski,S., & Szafałowicz, M. (2013). Temperature-dependent appearanceof forensically useful beetles on carcasses. Forensic ScienceInternational, 229(1-3), 92-99.doi:10.1016/j.forsciint.2013.03.034
Maurer,H. H. (2007). Current role of liquid chromatography–massspectrometry in clinical and forensic toxicology. Analyticaland bioanalytical chemistry, 388(7),1315-1325.
McIntyre,I. M. (2014). Liv forensicscience, medicine, and pathology, 10(1),91-96.
Meyer,M. R., Helfer, A. G., & Maurer, H. H. (2014). The currentposition of high-resolution MS for drug quantification in clinical &forensic toxicology. Bioanalysis, 6(17), 2275-2284.doi:10.4155/bio.14.164
Mozayani,A., & Noziglia, C. (Eds.). (2010). Theforensic laboratory handbook procedures and practice.Springer Science & Business Media.
Murthy,V., & Mohanty, M. (2010). Entomotoxicology: A review.
Muuka,G., Hang’ombe, B. M., Nalubamba, K. S., Kabilika, S., Mwambazi, L.,& Muma, J. B. (2011). Comparison of complement fixation test,competitive ELISA and LppQ ELISA with post-mortem findings in thediagnosis of contagious bovine pleuropneumonia (CBPP). TropicalAnimal Health and Production, 43(5),1057-1062.
Nagy,Z. T. (2010). A hands-on overview of tissue preservation methods formolecular genetic analyses. OrganismsDiversity & Evolution, 10(1),91-105.
NationalResearch Council. (2009). Strengtheningforensic science in the United States: a path forward.National Academies Press.
Ojanperä,I., Kolmonen, M., & Pelander, A. (2012). Current use ofhigh-resolution mass spectrometry in drug screening relevant toclinical and forensic toxicology and doping control. Analyticaland bioanalytical chemistry, 403(5),1203-1220.
Oliveira,T. C., & Vasconcelos, S. D. (2010). Insects (Diptera) associatedwith cadavers at the Institute of Legal Medicine in Pernambuco,Brazil: Implications for forensic entomology. ForensicScience International, 198(1),97-102.
Paczkowski,S., & Schütz, S. (2011). Post-mortem volatiles of vertebratetissue. Appliedmicrobiology and biotechnology, 91(4),917-935.
Peters,F. T. (2011). Recent advances in liquid chromatography–(tandem)mass spectrometry in clinical and forensic toxicology. ClinicalBiochemistry, 44(1),54-65.
Picard,C. J., & Wells, J. D. (2009). Survey of the genetic diversity ofPhormia regina (Diptera: Calliphoridae) using amplified fragmentlength polymorphisms. Journalof medical entomology, 46(3),664-670.
Pohjoismäki,J. L., Karhunen, P. J., Goebeler, S., Saukko, P., & Sääksjärvi,I. E. (2010). Indoors forensic entomology: colonization of humanremains in closed environments by specific species ofsarcosaprophagous flies. ForensicScience International, 199(1),38-42.
Primorac,D., & Schanfield, M. (Eds.). (2014). ForensicDNA Applications: An Interdisciplinary Perspective.CRC Press.
Saks,M. J., & Koehler, J. J. (2008). The individualization fallacy inforensic science evidence.
Schweitzer,N. J., & Saks, M. J. (2007). The CSI effect: popular fictionabout forensic science affects the public`s expectations about realforensic science. Jurimetrics,357-364.
Singh,J., & Sharma, B. R. (2008). Forensic entomology: a supplement toa forensic death investigation. JPAFMAT, 8(1),26-33.
Sisco,E., Forbes, T. P., Staymates, M. E., & Gillen, G. (2016). Rapidanalysis of trace drugs and metabolites using a thermal desorptionDART-MS configuration. AnalyticalMethods, 8(35),6494-6499.
Skopp,G. (2010). Postmortem toxicology. Forensicscience, medicine, and pathology, 6(4),314-325.
Sonet,G., Jordaens, K., Braet, Y., & Desmyter, S. (2012). Why is themolecular identification of the forensically important blowflyspecies Lucilia caesar and L. illustris (family Calliphoridae) soproblematic?. ForensicScience International, 223(1),153-159.
Sukontason,K., Piangjai, S., Siriwattanarungsee, S., & Sukontason, K. L.(2008). Morphology and developmental rate of blowflies Chrysomyamegacephala and Chrysomya rufifacies in Thailand: application inforensic entomology. ParasitologyResearch, 102(6),1207-1216.
Symes,S. A., Rainwater, C. W., Chapman, E. N., Gipson, D. R., & Piper,A. L. (2008). Patterned Thermal Destruction of Human Remains in aForensic Setting-2.
Tarone,A. M., & Foran, D. R. (2011). Gene expression during blow flydevelopment: improving the precision of age estimates in forensicentomology. Journalof forensic sciences, 56(s1).
Tomberlin,J. K., Mohr, R., Benbow, M. E., Tarone, A. M., & VanLaerhoven, S.(2011). A roadmap for bridging basic and applied research in forensicentomology. Annualreview of Entomology, 56,401-421.
Tsokos,M., & Byard, R. W. (2012). Putrefactive "rigor mortis."Forensicscience, medicine, and pathology, 8(2),200-201.
Vasconcelos,S. D., Soares, T. F., & Costa, D. L. (2013). Multiplecolonization of a cadaver by insects in an indoor environment: firstrecord of Fannia trimaculata (Diptera: Fanniidae) and Peckia (Peckia)chrysostoma (Sarcophagidae) as colonizers of a humancorpse. International Journal of Legal Medicine, 128(1),229-233. doi:10.1007/s00414-013-0936-2
Vass,A. A. (2011). The elusive universal post-mortem intervalformula. ForensicScience International, 204(1),34-40.
Verma,K. (2013). Effects of Codeine, Sodium Pentothal and DifferentTemperature Factors on the Growth Rate Development of Chrysomyarufifacies for the Forensic Entomotoxicological Purposes. JBioanal Biomed, 5,006-012.
Verma,K., & Paul, R. (2013). Assessment of post-mortem interval,(PMI)from forensic entomotoxicological studies of larvae andflies. EntomolOrnithol Herpetol, 2(104),2161-0983.
Villet,M. H., Richards, C. S., & Midgley, J. M. (2009). Contemporaryprecision, bias, and accuracy of minimum post-mortem intervalsestimated using development of carrion-feeding insects. In Currentconcepts in forensic entomology (pp.109-137). Springer Netherlands.
Wallace,M. (2011). New Frontiers in Molecular Forensics. ForensicScience Advances and Their Application in the Judiciary System,33.
Wells,J. D., & Stevens, J. R. (2008). Application of DNA-based methodsin forensic entomology. Annu.Rev. Entomol., 53,103-120.
Wielgomas,B., Czarnowski, W., & Jansen, E. H. (2012). Persistentorganochlorine contaminants in hair samples of Northern Polandpopulation, 1968–2009. Chemosphere, 89(8),975-981.
Zhou,L., Liu, L., Chang, L., & Li, L. (2011). Poisoning Deaths inCentral China (Hubei): A 10‐yearRetrospective Study of Forensic Autopsy Cases. Journalof forensic sciences, 56(s1).
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