Dr Shafiq Syed

Forensic science is the application of a broad spectrum of sciences and technologies to investigate situations after getting the facts and to establish what occurred based on collected evidence. This is especially important in law enforcement where forensics is done in relation to criminal or civil law. In civil actions, forensics can help resolve a broad spectrum of legal issues through the identification, analysis, and evaluation of physical evidence. The field of forensic science covers document examination, DNA analysis using electronic or digital media, fingerprinting, autopsy techniques, forensic engineering, forensic anthropology, pathology, economics, accounting, biology, entomology, toxicology, and much more. In this chapter we have described different materials such as hair, blood, bone, teeth, saliva, nails, feathers, skin, leather, sperm, feces, and urine and different methods for extracting DNA from different sources. The applications of animal forensics can be broadly viewed in the following four categories such as animals can be the victim, can be the perpetrator, and can be the witness and wildlife forensics. Molecular animal forensics provides different genetic tools such as DNA sequencing, single nucleotide polymorphism (SNP), PCR-RFLP, and microsatellite analysis for species identification and for characterization or identification of a sample recovered from a crime scene or illegal wildlife traders and black markets involved in wildlife trade. The genetic identification can be done as species identification, identification of geographic origin, individual identification, etc. Mitochondrial and nuclear markers can be used for genetic identification of the species. Identification of geographic origin is done by phylogeography and population assignment methods. To summarize, various techniques of individual identification, sexing, and parentage can be used. These techniques involve the microsatellite genotyping, DNA nucleotide sequencing, SNP typing, RAPD, and AFLP.

Bluetongue (BT) is an economically important viral disease of sheep and goat causing high morbidity and mortality. In the present study, eight Bluetongue virus (BTV) isolates from Utter Pradesh (UP), four BTV isolates from Andhra Pradesh (AP) and one isolate each from Haryana, Rajasthan and Tamil Nadu (TN) states were adapted in BHK- 21 cell line. All the isolates were confirmed as bluetongue virus (BTV) based on amplification of 366bp amplicon with group specific ns1 gene based RT-PCR. All the isolates were then confirmed as BTV serotype-1 based on segment 2 (vp2 gene) specific RT-PCR amplifying a specific amplicon of 605bp size. The isolates were further confirmed as BTV-1 by nucleotide sequencing of partially amplified products of vp2 gene. The sequencing data thus generated was analysed to seek for any intraserotypic variations among these various Indian isolates of BTV-1 which originated from different states of India. For this purpose, restriction map for various restriction enzymes (RE) was constructed after carrying out in silico restriction enzyme analysis using Restriction Mapper V 3.0. software. In silico restriction enzyme analysis (REA) with restriction enzyme (RE) HindII, of two samples NRT35 (JQ037810) and NRT39 (JQ037809) from AP revealed a single RE site at nt1250. One isolate each from UP (MKD25 (JQ037807)), Rajasthan (AY559058) and Haryana (AY559060) also revealed a single RE site at nt1531, whereas remaining isolates revealed presence of two RE sites at nt1250 and nt1531. In case of TN (AY559061) and BT1 (JQ037811) isolate, REA with RE TaqI revealed one extra RE site at nt1244 in addition to the one at nt1513 in remaining isolates, where as one isolate each from UP (MKD22 (JQ037804)) and AP (NRT35 (JQ037810)) revealed one additional RE site at nt1285 and nt1685 respectively, with RE NdeII. A little genetic diversity was observed within BTV1 isolates of adjoining states such as Haryana, Rajasthan, Andhra Pradesh and Tamil Nadu due to point mutations. The presence of a common culicoides vector species C. oxystoma has been reported in all these states and could be the possible cause for spread of virus in these parts of India. © 2013 by Nova Science Publishers, Inc. All rights reserved.

One of the most important vector borne diseases of ruminants worldwide is causedby bluetongue (BT) virus, an orbivirus of the Reoviridae family. Bluetongue (BT) is aneconomically important viral disease of ruminant species especially sheep and goatcausing high morbidity and mortality.The BT virus has segmented genome and is thus prone to frequent mutations andgene reassortment following the co-infection of two or more viruses in the host cell,leading to emergence of new genetic variants. For the present study, vp2 gene of Indianisolate KMNO-7 of BTV21, adapted in BHK-21 cell line, was sequenced full lengthutilising piecemeal approach after designing 7 primer pairs for the same and in silicorestriction enzyme analysis (REA) was done.The Restriction enzyme profile of the vp2 gene of the Indian isolate was thencompared with that of the other global isolates of BTV21 namely BTV21/ japan/TO2-1(AB686218.1), BTV21/japan/ ON89-1 (AB686215.1) and BTV21/SA-ref (AJ585142.1).Such comparison revealed certain restriction sites which were unique to the isolates andthus could help in differentiating Indian isolate from the global isolates. It was found thatthe restriction site for the enzyme EcoRV was present exclusively in the Indian isolateKMNO-7 while other isolates lacked the site for the enzyme. Similarly restriction site forthe enzyme Xbal1 was present exclusively in the BTV21/japan/ON89-1 isolate and not inthe other isolates. BTV21/SA-ref (AJ585142.1) lacked the restriction sites for theenzymes Sal1, Sca1 and BamH1 whereas other isolates possessed the restriction sites forthese enzymes. The restriction site for the enzyme Hind111 was absent inBTV21/japan/ON89-1isolate but was present in the remaining three isolates. Although there are various nucleotide sequence based methods for the differentiation of variousstrains of bluetongue virus, however, REA can serve as a quick method of differentiatingglobal isolates of a particular bluetongue virus serotype which can be furthercharacterised after deducing the nucleotide sequence of the vp2 gene. © 2013 by Nova Science Publishers, Inc. All rights reserved.

The epithelial lining of the fallopian tube is vital for fertility, providing nutrition to gametes and facilitating their transport. It is composed of two major cell types: secretory cells and ciliated cells. Interestingly, human ovarian cancer precursor lesions primarily consist of secretory cells. It is unclear why secretory cells are the dominant cell type in these lesions. Additionally, the underlying mechanisms governing fallopian tube epithelial homoeostasis are unknown. In the present study, we showed that across the different developmental stages of mouse oviduct, secretory cells are the most frequently dividing cells of the oviductal epithelium. In vivo genetic cell lineage tracing showed that secretory cells not only self-renew, but also give rise to ciliated cells. Analysis of a Wnt reporter mouse model and various Wnt target genes showed that the Wnt signaling pathway is involved in oviductal epithelial homoeostasis. By developing two triple-transgenic mouse models, we showed that Wnt/ ß-catenin signaling is essential for self-renewal as well as the differentiation of secretory cells. In summary, our results provide mechanistic insight into oviductal epithelial homoeostasis.

Organ shape and size are important determinants of their physiological functions. Epithelial tubes are anlagen of many complex organs. How these tubes acquire their complex shape and size is a fundamental question in biology. In male mice, the Wolffian duct (WD; postnatally known as epididymis) undergoes an astonishing transformation, where a straight tube only a few millimetres long elongates to over 1000 times its original length and fits into a very small space, due to extensive coiling of epithelium, to perform the highly specialized function of sperm maturation. Defective coiling disrupts sperm maturation and leads to male infertility. Recent work has shown that epithelial cell proliferation is a major driver of WD coiling. Still, very little is known about the molecular signals involved in this process. Testicular androgens are known regulators of WD development. However, epithelial androgen receptor signalling is dispensable for WD coiling. In this study, we have shown that Wnt signalling is highly active in the entire WD epithelium during its coiling, and is limited to only a few segments of the epididymis in later life. Pharmacological and genetic suppression of Wnt signalling inhibited WD coiling by decreasing cell proliferation and promoting apoptosis. Comparative gene expression analysis identified Fibroblast growth factor 7 (Fgf7) as a prime Wnt target gene involved in WD coiling and in vitro treatment with Fgf7 protein increased coiling of WDs. In summary, our work has established that epithelial canonical Wnt signalling is a critical regulator of WD coiling and its precise regulation is essential for WD/epididymal differentiation.

In order to detect polymorphism in exon 20 of the leptin receptor (LEPR) gene and its possible association with postpartum anestrus (PPA), peripheral blood samples were obtained from 40 Murrah buffaloes including 20 PPA (>120 days after parturition) and 20 normal cyclic (<60 days after parturition) buffaloes having similar postpartum estrous conditions over previous consecutive gestations. Genomic DNA was isolated, and PCR was standardized to amplify partial exon 20 of the LEPR gene of 413 bp. Amplified fragments of the gene were sequenced and sequence variation was detected by assessing multiple alignments. The LEPR gene showed polymorphism at A231G, C247A, and G347A. Chi-square test of these 3 polymorphism sites between the 2 groups did not reveal any significant association of polymorphism with PPA. Hence, polymorphism at exon 20 of the LEPR gene was found in Murrah buffaloes, but the role of these polymorphisms in PPA could not be established.

The genome of bluetongue virus (BTV) consists of 10 segments. Of these seg-2 encoded VP2 is the major serotype determining protein, and seg-6 encoded VP5 protein enhances the protective neutralizing activity of VP2 protein inducing higher serotype specific antibody titer than the VP2 alone. Out of the twenty-six BTV serotypes found worldwide, 22 were reported from different states of India. These include serotype 21 which was recently isolated from Andhra Pradesh, and was involved in a severe outbreak of bluetongue in Indian native sheep. BTV21 (KMNO-7) and BTV16 were circulating at the same time. This co-circulation, along with the fact that the virus genome is segmented, provides an opportunity for these two isolates of different serotypes to simultaneously infect the same animal, and even the same cell or a same vector with the potential for generation of reassortant viruses. This study was carried out to provide some insights into the outbreak. We carried out full length sequencing of genome seg-2 and seg-6 of Indian isolates VJW64 (BTV16) and KMNO-7 (BTV21). Detailed phylogenetic analysis revealed that genome seg-6 of Indian isolate KMNO-7 (BTV21) clusters with isolates of BTV16 showing maximum nucleotide similarity of 97.6% with TUR/2000/02 isolate of BTV16, which is much more than it shows with any isolate of BTV21. KMNO-7 (BTV21) significantly diverged from original strain of BTV21, and is a reassortant strain having acquired seg-6 from an isolate of BTV16. This study provides some useful insights into the epidemiology of the bluetongue disease, and undermines serotyping on genome seg-6 basis. © 2013 Elsevier B.V.