Wildlife crime suffers from low prosecution and conviction rates, with a lack of evidence and resources cited as hurdles to enforcement. Forensic evidence is used in human-on-human crimes to identify ...perpetrators and link individuals to criminal activity. Forensics approaches in the context of wildlife crime are heavily focused on non-human evidence using DNA barcoding to establish species and geographical origins. In human-on-human crime fingermarks and DNA profiling are two of the most recognisable forensic evidence types, both with significant global infrastructure, which contribute to prosecutions and convictions. Wildlife products can be the only physical evidence type available in a wildlife crime but attempts to recover human forensic evidence from them is a relatively unexplored area. The research that does exist demonstrates fingermark and touch DNA evidence can be collected in many contexts from several different species. Despite this there has been only one report of utilisation of this type human evidence recovery in wildlife case work. Failure to consider all potential evidence types has a negative impact on wildlife crime investigations. There is a need to experimentally assess the benefits and limitations associated with the collection of human evidence from wildlife items. This article introduces key factors that affect the recovery of human fingermarks and touch DNA evidence before focussing on the limited number of instances where these methods have been applied to wildlife forensic research and what considerations should be taken when developing further work in this field.
•Prosecution and convictions rates in wildlife crimes are low.•Low resources and lack of evidence hamper wildlife crime investigations.•Fingermarks and human touch DNA are two common forms of forensic evidence.•The recovery of human evidence in wildlife crime case work is limited.•The use of existing human forensic infrastructure deserves more consideration.
: A common limitation to most forensic trace evidence analysis is the ability to determine the time at which the evidence was deposited at the crime scene. This issue of timing is vitally important ...as it may not only reveal when the crime occurred, but could exclude potential suspects from the investigation. Using a reverse transcription quantitative polymerase chain reaction (RT‐qPCR) assay, we monitored the relative expression ratio (RER) of two different RNA species (18S and β‐actin) in hair samples that were aged naturally over a period of 3 months. No gender or age‐of‐donor biases were observed, and results were linear up to 60 days. After 60 days, the results were more variable and gave unreliable estimates of time since deposition. Overall, the results presented in this paper suggest that the age of hair samples containing follicular tags can be approximated using a second‐order polynomial, although with limitations: Age = 3.31RER2 – 2.85RER – 0.54 (R2 = 0.98).
•End-user assessment of first quantification kit to include both genomic and Y degradation metrics.•Data describes key performance criteria to support application in casework.•Assay can detect mixed ...sexual assault case type samples at quantification stage.•Assay can quantify DNA recovered from human teeth from the middle ages.
The Quantiplex® Pro RGQ kit quantifies DNA in a sample, supports the detection of mixtures and assesses the extent of DNA degradation based on relative ratios of amplified autosomal and male markers. Data show no significant difference in the accuracy and sensitivity of quantification between this and the Promega PowerQuant® System, both detecting the lowest amount of DNA tested, 4 pg. Laboratory controlled mixed male:female DNA samples together with mock sexual assault samples were quantified across a range of mixture ratios. Analysis software detected mixed DNA samples across all ratios for both quantification kits. Subsequent STR analysis using the Investigator® 24Plex QS Kit was able to corroborate mixture detection down to 1:25 male:female DNA ratios, past which point mixtures appeared identical to single-source female samples. Analysis software also detected laboratory degraded DNA samples, with data showing a positive trend between the Degradation Index (DI) and length of time of sonication. When used on ancient remains the assay was able to triage samples for further analysis, and STR profiles were concordant with DNA quantification results in all instances. STR analyses of laboratory-controlled sensitivity, mixture, and degradation studies supports the quality metric obtained from quantification. These data support the use of the Quantiplex® Pro RGQ kit for sample screening and quantification in forensic casework and ancient DNA studies.
The 16S-23S ribosomal DNA spacer region of selected cyanobacterial strains was amplified by the polymerase chain reaction using primers to conserved flanking sequences. Single or multiple rDNA ...amplification products were generated depending on the strain and primer pair. Species could generally be distinguished on the basis of size heterogeneity of the products. Analysis of restriction digests of the amplified rDNAs indicated polymorphisms useful in identification. Four enzymes (
HinfI,
DdeI,
AluI,
TaqI) generated restriction fragment length patterns that could discriminate between the cyanobacteria to the taxonomic levels of genus and species. This approach should prove useful in the rapid identification of cyanobacteria.
• The effect is reported here of cytochalasin E isolated from the fungus Rosellinia necatrix on photosynthesis in young leaves of Malus domestica (apple). • Cytochalasin E was administered via the ...petiole to excised leaves. The chlorophyll fluorescence emission spectrum and time resolved fluorescence decay were measured up to the point where visible leaf damage was observed. • Within 2 h, the ratio of fluorescence emission at 730 nm decreased with respect to the peak at 690 nm. Over 6 h a small blue shift in the 690 nm emission band to 685 nm was seen. The time resolved fluorescence decay showed changes over a similar timescale after administration of cytochalasin E. The control decay could be fitted by two components, τ1, 112 ps, τ2, 402 ps, but after 6 h treatment with cytochalasin E the decay required a further component τ3, 4.25 ns for a good fit. • Cytochalasin E has a direct effect on photosynthesis, possibly as a result of impairment of light harvesting. This might partially account for the pathogenicity of the root infecting R. necatrix. Fluorescence techniques may therefore provide an early detection system for the fungus, a necessary prerequisite for development of a control strategy.
Abstract
The 16S-23S ribosomal DNA spacer region of selected cyanobacterial strains was amplified by the polymerase chain reaction using primers to conserved flanking sequences. Single or multiple ...rDNA amplification products were generated depending on the strain and primer pair. Species could generally be distinguished on the basis of size heterogeneity of the products. Analysis of restriction digests of the amplified rDNAs indicated polymorphisms useful in identification. Four enzymes (Hin fI, Dde I, Alu I, Taq I) generated restriction fragment length patterns that could discriminate between the cyanobacteria to the taxonomic levels of genus and species. This approach should prove useful in the rapid identification of cyanobacteria.
The main objective of this work was to characterize cyanobacterial PS2 polypeptides with particular respect to calcium binding. A number of cyanobacteria were studied by trypsin digestion of their ...thylakoid membranes which showed that using the same method of preparation some cyanobacteria produce everted thylakoid vesicles whilst others produce right sided vesicles. No direct correlation between the sidedness of the produced thylakoids and any growth or metabolic characteristic of the organithm was found. In Synechocystis 6308 a polypeptide of 27kDa , apparently unrelated to known polypeptides of the oxygen evolving complex, was shown to disappear on trypsin treatment, a disappearance which correlated to the loss of oxygen evolving activity. Calcium binding studies performed on Synechococcus 7942 showed the presence of a calmodulin-like calcium binding site within both thylakoids and PS2. The protein on which this site resided was not proven but results pointed to the involvement of the Dl protein. Synechococcus 7942 produces two forms of the Dl protein and mutants in which one form was deleted showed differing affinities for calcium both in vivo (during growth) and in vitro (during reactivation experiments). Differences were also noted for spectra from the mutant strains with one form appearing to retain greater quantities of phycocyanin on the thylakoid membranes. Protein kinase assays performed on the Dl mutants also showed differences between these and the wild type organism. These differences were noted for small polypeptides of 8, 10 and 12kDa. A phosphoprotein of 14kDa was shown to be related to the psb H gene product and a phosphoprotein of approximately 32kDa, possibly Dl, was also obtained. A further 58kDa phosphoprotein which may be related to the calmodulin site or to the protein kinase responsible for PS2 phosphorylation was noted. Both calcium and magnesium were shown to have specific effects on phosphorylation primarily with respect to the small polypeptides mentioned above. Studies on the possible glycosylation of P52 and thylakoid polypeptides produced evidence to suggest that either the Dl or D2 polypeptides are glycosylated and that this may provide a mechanism by which the PS2 complex is held together. Evidence in support of glycosylation of phycobilisome components was also produced with polypeptides of 34.5, 45 78 and 90kDa appearing to be glycosylated. A polypeptide of 50kDa which is closely associated with PS2 was shown to be related , in terms of homology, to a 20kDa DCCD-binding protein of plants which is thought to be a LHC protein.
The main objective of this work was to characterize cyanobacterial PS2 polypeptides with particular respect to calcium binding. A number of cyanobacteria were studied by trypsin digestion of their ...thylakoid membranes which showed that using the same method of preparation some cyanobacteria produce everted thylakoid vesicles whilst others produce right sided vesicles. No direct correlation between the sidedness of the produced thylakoids and any growth or metabolic characteristic of the organithm was found. In Synechocystis 6308 a polypeptide of 27kDa , apparently unrelated to known polypeptides of the oxygen evolving complex, was shown to disappear on trypsin treatment, a disappearance which correlated to the loss of oxygen evolving activity. Calcium binding studies performed on Synechococcus 7942 showed the presence of a calmodulin-like calcium binding site within both thylakoids and PS2. The protein on which this site resided was not proven but results pointed to the involvement of the Dl protein. Synechococcus 7942 produces two forms of the Dl protein and mutants in which one form was deleted showed differing affinities for calcium both in vivo (during growth) and in vitro (during reactivation experiments). Differences were also noted for spectra from the mutant strains with one form appearing to retain greater quantities of phycocyanin on the thylakoid membranes. Protein kinase assays performed on the Dl mutants also showed differences between these and the wild type organism. These differences were noted for small polypeptides of 8, 10 and 12kDa. A phosphoprotein of 14kDa was shown to be related to the psb H gene product and a phosphoprotein of approximately 32kDa, possibly Dl, was also obtained. A further 58kDa phosphoprotein which may be related to the calmodulin site or to the protein kinase responsible for PS2 phosphorylation was noted. Both calcium and magnesium were shown to have specific effects on phosphorylation primarily with respect to the small polypeptides mentioned above. Studies on the possible glycosylation of P52 and thylakoid polypeptides produced evidence to suggest that either the Dl or D2 polypeptides are glycosylated and that this may provide a mechanism by which the PS2 complex is held together. Evidence in support of glycosylation of phycobilisome components was also produced with polypeptides of 34.5, 45 78 and 90kDa appearing to be glycosylated. A polypeptide of 50kDa which is closely associated with PS2 was shown to be related , in terms of homology, to a 20kDa DCCD-binding protein of plants which is thought to be a LHC protein.
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