Shed Hairs Research Articles

Triaging of hair samples using fluorescent in situ detection

ABSTRACT DiamondTM Nucleic Acid Dye (DD) has been evaluated for visualizing DNA material on various evidence types, including nuclei in plucked hair roots. While plucked hair roots exhibit a rapid DNA visualization process, shed hairs, even when stained with DD, pose challenges due to the lack of cell nuclei. In the absence of clear cell nuclei, the sole indicator of DNA presence is the fluorescent signal from DD bound to DNA. This study aimed to assess the stability of the fluorescent signal in shed hairs, revealing a rapid decline within 2–3 min post-staining, likely attributed to the cell-free nature of the DNA present on the hair root. Consequently, immediate microscopic visualization and imaging post-staining are crucial during shed hair examination.

Evaluation of the relationship of digital phototrichogram findings of patients with diffuse hair loss with blood TSH, ferritin and vitamin B12 levels.

Telogen effluvium (TE) is a type of alopecia that is frequently seen in women. Among factors resulting in hair loss, many reasons such as endocrine diseases, nutrition disorders, stress, anemia, low ferritin levels, vitamin B12 deficiency, and thyroid diseases are found. A digital phototrichogram is one of the non-invasive methods of diagnosis in the evaluation of alopecia. In this study, it was aimed to compare biochemical parameters of female patients with diffuse hair loss with phototrichogram findings. 108 female patients with diffuse hair loss were included in the study. Patients were divided into two groups: acute and chronic TE, and a hair pull test was applied. Total blood count, blood biochemistry, iron, iron binding capacity, ferritin, thyroid stimulating hormone (TSH), sT3, sT4, folic acid, and vitamin B12 levels were examined. The telogen/anagen ratios and hair densities of patients were determined with a phototrichogram. The serum biochemical parameters (aspartate aminotransferase, alanine aminotransferase, urea, creatinine), TSH, sT3, sT4, and folic acid levels of patients were within normal limits. Telogen ratio, anagen ratio, hair density, number of shed hairs, family history, blood ferritin, TSH, and vitamin B12 levels were found to be similar between groups having acute and chronic TE. In our study, while mean anagen and telogen ratios with trichoscan were similar to literature data, no statistically significant correlation was determined between patients' ages and examined trichoscan findings (p>0.05). The hair pull test positivity of patients with chronic TE was higher compared to patients with acute TE (p<0.05). In patients with positive hair pull tests, the telogen ratio and hair density were found to be higher. In the group with <40 ng/mL ferritin level, the mean telogen ratio was detected to be significantly higher than the mean anagen ratio (p<0.05). No significant correlation was determined between vitamin B12 and TSH levels in patients and phototrichogram findings (p>0.05). The findings of this study showed that ferritin has an important role in diffuse hair loss, and the phototrichogram method is an auxiliary method for the physician in the diagnosis of TE.

An evaluation of the RapidHIT™ ID system for hair roots stained with Diamond™ Nucleic Acid Dye

The RapidHIT™ ID (RHID) system was evaluated for its suitability in processing a single hair root to obtain informative DNA profiles. Hair samples were assessed for nuclear DNA prior to DNA analysis using Diamond™ Nucleic Acid Dye (DD) and real-time Extended Depth of Field (EDF) imaging to visualise and count nuclei if present. Hairs were viewed under an Optico N300F LED Fluorescent Microscope and imaged using a MIchrome 5 Pro camera. Hair roots were processed through both the ACE GlobalFiler™ Express sample cartridge and the RapidINTEL™ sample cartridge. A total of 44 hairs including shed hairs (9) and plucked hairs (35) from 8 donors were evaluated in this study. The processing of hairs using the RHID system required the modification of a standard swab that allowed for hairs to be easily collected and placed into the cartridge but also allowed for the re-collection of hair roots post RHID analysis (for potential standard DNA workflow). 90% of plucked hairs with a high nuclei count (>100) resulted in a high partial or full DNA profile, with the remaining 10% resulting in a low partial profile. 44% of shed hairs resulted in a low partial profile, with the remaining hairs resulting in a null profile. This study demonstrated that the RHID system could successfully obtain a DNA profile from a single hair root with nuclei present post-DD staining. According to these results, it is suggested that when dealing with hairs containing fewer than 50 nuclei, using the RapidINTEL™ cartridge can enhance allele recovery.

197 Trace Mineral Source Affects Haircoat Characteristics and Shedding in Cats

Abstract Trace minerals have a pivotal role in maintaining skin health and haircoat characteristics in companion animals. The objective of the current study was to evaluate the effects of inorganic versus amino acid complexed sources of trace minerals on feline skin and haircoat characteristics as well as hair growth. A total of 40 cats (age 5 – 12 years; 22 female, 18 male) were used in this 105-d (15-d acclimation; 90-d test period) trial at Summit Ridge Farms (Susquehanna, PA) from January to April 2022. All cats were fed a basal diet that contained sulfate sources of trace minerals formulated to meet AAFCO (2021) recommendations for 15 d before being randomly assigned (blocked by haircoat color, age, sex, and weight) to 1 of 2 treatments. Dietary treatments included sulfate sources (CON; n = 20) and amino acid complexed (Zinpro ProPath, Zinpro Corp. Eden Prairie, MN) sources (ZPM; n = 20) of supplemental trace minerals. Both treatments were formulated to provide equal amounts of supplemental trace minerals (DM basis), differing only in source (100 ppm Zn, 15 ppm Mn, 12 ppm Cu, 80 ppm Fe). On d 15, a side patch was shaved just below the back (3” x 4”) to evaluate hair growth on d 60 and 105 (average of 3 measurements). Additionally, visual haircoat evaluations and shed hair amount were evaluated at d 60 and 105. All research personnel were blinded to treatments. Statistical analyses were completed using GLIMMIX procedure of SAS 9.4 with d 15 as a covariate and cat as the experimental unit. Trace mineral source did not affect body condition score at either time point. There was no treatment or treatment x day interaction for hair length. A trend (p = 0.13) for a 3-way interaction between treatment, sex, and day was detected due to male cats having longer hair at d 105. Shed hair was evaluated by brushing (20 strokes) and weighing collected hair. No overall treatment effect was detected for shed hair; although, a treatment x day interaction (P = 0.02) for shed hair was observed. At d 60, ZPM cats had 23 mg less shed hair compared with CON cats (P = 0.08). Haircoat scores for shine, scale, and overall quality were assessed by a veterinarian. There were no treatment or treatment x day interactions for shine or overall scores. A treatment x day interaction was detected (P = 0.02) for scale (dander). At d 60, ZPM cats had a reduced average scale score compared with CON cats (0.66 vs 1.05, respectively; P = 0.03). Overall, our data suggests trace mineral source may influence shed hair amount and dander production in cats. Further research to evaluate the influence of season and sex is warranted.

Citizen science facilitates first ever genetic detection of wolf-dog hybridization in Indian savannahs.

Human demographic expansion has confined wildlife to fragmented habitats, often in proximity to human-modified landscapes. Such interfaces facilitate increased interactions between feral or domesticated animals and wildlife, posing a high risk to wild species. This is especially relevant for free-ranging dogs (Canis lupus familiaris) and wild canids like gray wolves (Canis lupus) and golden jackals (Canis aureus). Wolf-dog hybridization may lead to a significant reduction of specific adaptations in wolves that could result in the decline of wolf populations. Detection and genetic discrimination of hybrids between dogs and wolves are challenging because of their complex demographic history and close ancestry. Citizen scientists identified two phenotypically different-looking individuals and subsequently collected non-invasive samples that were used by geneticists to test wolf-dog hybridization. Genomic data from shed hair samples of suspected hybrid individuals using double-digest restriction-site-associated DNA (ddRAD) sequencing resulted in 698 single nucleotide polymorphism (SNP) markers. We investigated the genetic origin of these two individuals analyzed with genetically known dogs, wolves, and other canid species including jackals and dholes (Cuon alpinus). Our results provide the first genetic evidence of one F2 hybrid and the other individual could be a complex hybrid between dogs and wolves. Our results re-iterate the power of next-generation sequencing (NGS) for non-invasive samples as an efficient tool for detecting hybrids. Our results suggest the need for more robust monitoring of wolf populations and highlight the tremendous potential for collaborative approaches between citizens and conservation scientists to detect and monitor threats to biodiversity.

Past Connectivity but Recent Inbreeding in Cross River Gorillas Determined Using Whole Genomes from Single Hairs.

The critically endangered western gorillas (Gorilla gorilla) are divided into two subspecies: the western lowland (G. g. gorilla) and the Cross River (G. g. diehli) gorilla. Given the difficulty in sampling wild great ape populations and the small estimated size of the Cross River gorilla population, only one whole genome of a Cross River gorilla has been sequenced to date, hindering the study of this subspecies at the population level. In this study, we expand the number of whole genomes available for wild western gorillas, generating 41 new genomes (25 belonging to Cross River gorillas) using single shed hairs collected from gorilla nests. By combining these genomes with publicly available wild gorilla genomes, we confirm that Cross River gorillas form three population clusters. We also found little variation in genome-wide heterozygosity among them. Our analyses reveal long runs of homozygosity (>10 Mb), indicating recent inbreeding in Cross River gorillas. This is similar to that seen in mountain gorillas but with a much more recent bottleneck. We also detect past gene flow between two Cross River sites, Afi Mountain Wildlife Sanctuary and the Mbe Mountains. Furthermore, we observe past allele sharing between Cross River gorillas and the northern western lowland gorilla sites, as well as with the eastern gorilla species. This is the first study using single shed hairs from a wild species for whole genome sequencing to date. Taken together, our results highlight the importance of implementing conservation measures to increase connectivity among Cross River gorilla sites.

Pitfalls, challenges and caveats in whole mitochondrial genome sequencing from hair shafts by MPS: Where, when and how to address them

The growing use of massively parallel sequencing (MPS) for the whole mitochondrial genome analysis in forensic laboratories, requires the establishment of efficient workflows and interpretation procedures, to support the feasibility of the technology and the reliability of the data. In the case of reference samples, such as blood and buccal swabs, the generation of mtDNA profiles by using MPS is relatively simple. Conversely, many forensic casework samples still pose challenges for the MPS, data interpretation and reporting of mtDNA. This is especially true for the analysis of shed hairs, which are one of the most common evidence types and which are among the most limited in terms of DNA quantity and quality. Due to these limitations, every step involved in the analysis become essentials and should be performed in order to obtain the best performance, optimizing the outcomes and minimizing the errors.In light of this, we present a study focusing on the extraction, quantification and MPS of the whole mtDNA in hair shafts, with the aims of set-up and validate a methodological pipeline to obtain the best sequencing results. The overall performance of the MPS panel, mainly in terms of total coverage, amplicons coverage and different primer pools efficiency, was evaluated also in relation to the different hair fragments, the mtDNA copy number used for libraries preparation and its degradation state.

Using Environmental Sampling to Enable Zoonotic Pandemic Preparedness

The current pandemic caused by the SARS CoV-2, tracing back its origin possibly to a coronavirus associated with bats, has ignited renewed interest in understanding zoonotic spillovers across the globe. While research is more directed towards solving the problem at hand by finding therapeutic strategies and novel vaccine techniques, it is important to address the environmental drivers of pathogen spillover and the complex biotic and abiotic drivers of zoonoses. The availability of cutting-edge genomic technologies has contributed enormously to preempt viral emergence from wildlife. However, there is still a dearth of studies from species-rich South Asian countries, especially from India. In this review, we outline the importance of studying disease dynamics through environmental sampling from wildlife in India and how ecological parameters of both the virus and the host community may play a role in mediating cross-species spillovers. Non-invasive sampling using feces, urine, shed hair, saliva, shed skin, and feathers has been instrumental in providing genetic information for both the host and their associated pathogens. Here, we discuss the advances made in environmental sampling protocols and strategies to generate genetic data from such samples towards the surveillance and characterization of potentially zoonotic pathogens. We primarily focus on bat-borne or small mammal-borne zoonoses and propose a conceptual framework for non-invasive strategies to tackle the threat of emerging zoonotic infections.

Going Bald — The Hairy Affair of Timing in Telemetry Studies: Moulting Activity in European Bat Species

Telemetry is a widely used method for radio-tracking bats. To minimize negative effects of attaching a tag, like bald spots during hibernation and nursery period, narrowing down the timing of the moult of bats is important. In this study, we investigated the moulting of 11 European bat species with a focus on Myotis daubentonii, Myotis myotis and Pipistrellus pipistrellus. Telemetry data were taken from a 20 years data base. For moulting activity bats were caught with mist nests during their activity period in 2019 and 2020. During data acquisition the condition of the hair was categorized using two different methods: visual hair growth and shed hair pulled off the bats' pelage. Both methods revealed the same results. In general, bats moulted once a year in mid-June (first moult: 17.06) until mid-August (last moult: 13.08), with a significant difference between adult males, reproductive females and non-reproductive females. Adult males (n = 197) and non-reproductive females (n = 60) moulted from mid-June (first moult: 17.06) to end of July (last moult: 27.07), while reproductive females (n = 216) moulted from beginning of July (first moult: 02.07) to mid-August (last moult: 17.08). This variance is probably caused by sex-specific energetic processes like pregnancy, lactation and spermatogenesis. When tagged before moulting season bats could regrow fur in the same year. As seen in recaptured bats (n = 31), tagged in or after this period bats stayed bald during hibernation and reproduction period until the following moulting season. Further studies are needed to specifically assess the impact of bald spots on hibernation and reproductive success. A preceding cost-benefit analysis of the expected knowledge gain versus the potential negative impact on individual and colony scale is recommended.

Hair Shedding Evaluation for Alopecia: A Refined Wash Test.

PurposeThe aim of this study is to establish a refined wash test (RWT) and explore the characteristics of daily shedding hair parameters in normal individuals and patients with androgenetic alopecia (AGA) or telogen effluvium (TE).Materials and MethodsChinese subjects diagnosed as normal, AGA or TE were enrolled. For the RWT procedure, the subjects were asked to pre-wash their hair 24 hours before the first collection, then to wash and rinse their hair with flowing water above a sink covered with a piece of filtering cloth. All hairs entrapped in the filtering cloth were air dried and collected. This “wash-and-collect” process was repeated for 3 consecutive days with a 24 ± 2 hour interval. Collected hairs were counted manually, and hair diameter was measured by videodermoscope. Number and diameter of daily shedding hairs and percentage of vellus and terminal hairs were analyzed.ResultsA total of 176 subjects participated in and completed the study. Shedding hair number during daily shampooing was 27.9 ± 12.2, 52.2 ± 28.5 and 125.5 ± 62.7 hairs in normal, AGA and TE participants, respectively, and the differences between each two of the three groups were all statistically significant. Diameter of shedding hairs was 76.9 ± 9.0 μm, 60.0 ± 9.5 μm and 82.7 ± 9.2 μm in normal, AGA and TE, respectively. Along with advancement of AGA severity, shedding hair number was generally increased, while hair diameter decreased. The percentage of vellus hairs in AGA was significantly higher than in the normal and TE groups (8.3 ± 6.6% vs 1.0 ± 1.6% and 1.0 ± 1.0%). The combination of shedding hair number and percentage of vellus hairs had a high diagnostic confidence (AUC = 0.957) to diagnose AGA from normal. Shedding hair number alone showed high confidence to diagnose TE from normal (AUC = 0.999).ConclusionThe non-invasive, easy-to-use RWT established in this study is a valuable tool to evaluate daily shedding hairs, which may be useful in diagnosis, dynamic monitoring of disease progression and treatment efficacy.

Detection of deletion/insertion polymorphism profiles from single human hair shafts.

Hair is a frequently encountered biological evidence in personal identification. The amount of nuclear DNA that can be extracted from a single strand of rootless hair is most limited, making the detection of short tandem repeat (STR) polymorphisms difficult. To overcome these limitations, deletion/insertion polymorphisms (DIP) as a new type of genetic marker have shown their benefits in detecting low-copy-number DNA. The Investigator DIPplex kit contains 30 biallelic autosomal DIP and amelogenin. The analysis of DIPs combines the advantages of both STR and single nucleotide polymorphism analyses. Thus, this study aimed to detect the DIP distribution of individual hair shafts from individuals. DNA was extracted from the shaft of fresh, aged, and shed hair. After DNA was evaluated, the DIP profiles were detected by capillary electrophoresis. The results indicated that the amount of DNA extracted from hair roots was much higher than that from the hair shafts in the same individual for all samples. The degradation index values of DNA from the aged hair shafts were highest. It is classified to be "mildly degraded." Compared with their hair roots, the full DIP profiles were detected for fresh hair, 70% for aged hair, and 92% for shed hair. Contrarily, except for fresh hair shafts, only three STR loci of the aged and shed strands of hair could be genotyped using AmpFlSTR MiniFiler PCR Amplification Kit. These results indicate that the detection of DIP profile is an effective tool for personal identification from hair shafts, including aged hair.

Comparison of polymerases used for amplification of mitochondrial DNA from challenging hairs and hairs of various treatments

Forensic DNA analysis of hair evidence typically involves the amplification and sequencing of the control region (CR) of the mitochondrial genome (mtgenome). In compromised hair samples, such as shed hairs, the number of mtgenome copies could be low; thus, it is imperative that the polymerase used in PCR is efficient to ensure maximum amplification. Considering this, the first phase of this study compared the yields obtained from 12 polymerases (sourced from a range of commercial companies) when amplifying the CR, hypervariable (HV) region II (HV2), and hypervariable subregion II-B (HV2B). This initial assessment was performed using mitochondrial DNA (mtDNA) extracted from 2 cm of hair adjacent to the root from three donors of different self-reported ancestries and hair color/texture. PrimeSTAR HS and KAPA HiFi HotStart consistently generated significantly higher amplicon yields (p < 0.05, ~5-fold increase) for most regions than AmpliTaq Gold DNA polymerase (the polymerase validated for use in most forensic laboratories). The second phase of this project was focused on assessing the broad utility of these top two performing polymerases for amplifying two regions of the mtgenome (CR and HV2B) from hair samples representing diverse self-reported ancestral origins (European, Latin American, African American, Asian, and Native American), characteristics/treatments (bleached, dyed, and chemically straightened), and anatomical origins (e.g., head and pubic region) (n = 41). These regions were chosen as they are the most challenging to amplify and sequence in compromised hair samples due to length (i.e., the CR is ~1.2 kb) and repeat structure (i.e., the polycytosine stretch within HV2B). The results indicated that regardless of sample type, PrimeSTAR HS and KAPA HiFi HotStart polymerases outperformed (p < 0.05) AmpliTaq Gold DNA polymerase (averaging 11- and 8-fold increased yields, respectively). The results from this study highlight that enhanced commercially available polymerases appear to significantly improve the amplification of mtDNA from challenging hair samples.

Forensic determination of hair deposition time in crime scenes using electron paramagnetic resonance.

Several types of biological samples, including hair strands, are found at crime scenes. Apart from the identification of the value and the contributor of the probative evidence, it is important to prove that the time of shedding of hair belonging to a suspect or victim matches the crime window. To this end, to estimate the ex vivo aging of hair, we evaluated time-dependent changes in melanin-derived free radicals in blond, brown, and black hairs by using electron paramagnetic resonance spectroscopy (EPR). Hair strands aged under controlled conditions (humidity 40%, temperature 20-22°C, indirect light, with 12/12hour of light/darkness cycles) showed a time-dependent decay of melanin-derived radicals. The half-life of eumelanin-derived radicals in hair under our experimental settings was estimated at 22±2days whereas that of pheomelanin was about 2days suggesting better stabilization of unpaired electrons by eumelanin. Taken together, this study provides a reference for future forensic studies on determination of degradation of shed hair in a crime scene by following eumelanin radicals by utilizing the non-invasive, non-destructive, and highly specific EPR technique.

Who killed my dog? Use of forensic genetics to investigate an enigmatic case

Genetic testing of animal biological material has become a valuable tool in forensic investigations, and it is successfully used to identify unknown crime perpetrators, to unmask food frauds, or to clarify cases of animal attacks on humans or other animals. When DNA profiling is not possible due to inadequate amounts of nuclear DNA, mitochondrial DNA (mtDNA) testing is the only viable alternative, as in the case of shed hair samples. In this case, a dog was allegedly killed by wild animals while being hosted in a boarding house. Extraneous hair fragments recovered from the dog’s mouth and paws were subjected to genetic analysis: the cytochrome b gene located on mtDNA was amplified and sequenced in order to determine the species responsible for the killing. The mtDNA analysis provided evidence that the dog was killed by other dogs, thus unmasking a false wild animal attack and putting the case in an entirely different perspective.

Surface-enhanced Raman Scattering and X-ray Fluorescence Analyses of a Single Hair Colored with a Hair Dye Product.

This paper describes a method for obtaining information that can contribute to individual identification from a single hair colored with a hair dye product using a combination of surface-enhanced Raman scattering (SERS) and X-ray fluorescence (XRF) analyses. SERS and XRF spectra of single hog hairs colored with several commercially available hair dye products were measured. SERS spectral patterns tended to be different depending on the hair dye products used for hair coloring. However, SERS spectral patterns of single hog hairs colored with different hair dye products that would produce similar types of dyes were similar. By performing XRF analysis, characteristic metallic elements originating from some hair dye products were detected. Therefore, XRF can contribute to identifying the difference among colored hairs that cannot be identified only by SERS. SERS and XRF analyses of a single shed hair can contribute to individual identification used in forensic science.

Are shed hair genomes the most effective noninvasive resource for estimating relationships in the wild?

Knowledge of relationships in wild populations is critical for better understanding mating systems and inbreeding scenarios to inform conservation strategies for endangered species. To delineate pedigrees in wild populations, study genetic connectivity, study genotype‐phenotype associations, trace individuals, or track wildlife trade, many identified individuals need to be genotyped at thousands of loci, mostly from noninvasive samples. This requires us to (a) identify the most common noninvasive sample available from identified individuals, (b) assess the ability to acquire genome‐wide data from such samples, and (c) evaluate the quality of such genome‐wide data, and its ability to reconstruct relationships between animals within a population.We followed identified individuals from a wild endangered tiger population and found that shed hair samples were the most common compared to scat samples, opportunistically found carcasses, and opportunistic invasive samples. We extracted DNA from these samples, prepared whole genome sequencing libraries, and sequenced genomes from these.Whole genome sequencing methods resulted in between 25%–98% of the genome sequenced for five such samples. Exploratory population genetic analyses revealed that these data were free of holistic biases and could recover expected population structure and relatedness. Mitochondrial genomes recovered matrilineages in accordance with long‐term monitoring data. Even with just five samples, we were able to uncover the matrilineage for three individuals with unknown ancestry.In summary, we demonstrated that noninvasive shed hair samples yield adequate quality and quantity of DNA in conjunction with sensitive library preparation methods, and provide reliable data from hundreds of thousands of SNPs across the genome. This makes shed hair an ideal noninvasive resource for studying individual‐based genetics of elusive endangered species in the wild.

Fragmented Nuclear DNA is the Predominant Genetic Material in Human Hair Shafts.

While shed hairs are one of the most commonly encountered evidence types, they are among the most limited in terms of DNA quantity and quality. As a result, nuclear DNA short tandem repeat (STR) profiling is generally unsuccessful and DNA testing of shed hair is instead performed by targeting the mitochondrial DNA control region. Although the high copy number of mitochondrial DNA relative to nuclear DNA routinely permits the recovery of mitochondrial DNA (mtDNA) data in these cases, mtDNA profiles do not offer the discriminatory power of nuclear DNA profiles. In order to better understand the total content and degradation state of DNA in single shed hairs and assess the feasibility of recovering highly discriminatory nuclear DNA data from this common evidence type, high throughput shotgun sequencing was performed on both recently collected and aged (approximately 50-year-old) hair samples. The data reflect trends that have been demonstrated previously with other technologies, namely that mtDNA quantity and quality decrease along the length of the hair shaft. In addition, the shotgun data reveal that nuclear DNA is present in shed hair and surprisingly abundant relative to mitochondrial DNA, even in the most distal fragments. Nuclear DNA comprised, at minimum, 88% of the total human reads in any given sample, and generally more than 95%. Here, we characterize both the nuclear and mitochondrial DNA content of shed hairs and discuss the implications of these data for forensic investigations.

Sequential cyclic changes of hair roots revealed by dermoscopy demonstrate a progressive mechanism of diffuse alopecia areata over time.

Diffuse alopecia areata (DAA) often leads to a complete hair shedding within a few months. To explore features and mechanisms underlying DAA. Scalp and hair root dermoscopy were conducted on 23 DAA patients throughout the disease process, 20 patchy Alopecia areata patients, 23 acute telogen effluvium (ATE) patients and 10 normal controls. Histopathology was also evaluated. We found almost all hair roots were anagen in early stage DAA in 18 patients (18/23, 78.3%) within the first 4-8weeks after hair loss onset. Anagen effluvium (~4weeks) was followed by catagen (~4weeks) and then telogen/exogen (~8weeks) effluvium with overlap. Hair root and proximal hair shaft depigmentation was more prominent in later DAA disease stages. Black dots, exclamation mark hairs and inconsistent thickness of hair shafts were found more often in early than later DAA (Ps<0.01). Early DAA histopathology revealed more prominent inflammation and hair follicle regression than that observed in the later stages. Patchy alopecia areata patients showed mixed anagen, catagen and telogen hair roots while ATE patients showed increased exogen and mildly decreased hair root pigmentation. Sequential cyclic staging of shed hairs in DAA indicates the insult may be hair-cycle specific. We suggest that DAA is initially an anagen effluvium disease involving an intense inflammatory insult, later progressing to a brief catagen effluvium, and then to telogen effluvium with premature exogen, in later stages of DAA.

068 Hair Shedding Score and Hair Coat Color Affect Body Temperature during Heat Stress in Weaned Beef Heifers

068 Hair Shedding Score and Hair Coat Color Affect Body Temperature during Heat Stress in Weaned Beef Heifers

Forensic analysis of mitochondrial control region DNA from single cat hairs

Although different parts of the mitochondrial control region (CR) have been described as forensically relevant, factors such as geographical location and breed may influence their forensic value. Analysis of multiple CR regions has been shown to increase the evidential value for certain populations, but due to large amplicon size, PCR success could be influenced when single shed hairs are used for analysis.A new strategy was developed to circumvent amplification of the repeat element RS2 without dramatically decreasing the number of potentially informative nucleotide positions sequenced bidirectional.DNA-extracts of 40 shed cat hairs were typed to compare the PCR success of this duplex strategy to the single amplicon approach. Amplicons were obtained from all tested hairs using the duplex strategy, compared to 60% using the single amplicon, illustrating the applicability of this strategy to case type samples.