A canine model to evaluate the effect of exercise intensity and duration on olfactory detection limits: the running nose.

Detection canines serve critical roles to support the military, homeland security and border protection. Some explosive detection tasks are physically demanding for dogs, and prior research suggests this can lead to a reduction in olfactory detection sensitivity. To further evaluate the effect of exercise intensity on olfactory sensitivity, we developed a novel olfactory paradigm that allowed us to measure olfactory detection thresholds while dogs exercised on a treadmill at two different exercise intensities. Dogs (n = 3) showed a decrement in olfactory detection for 1-bromooctane at 10-3 (v/v) dilutions and lower under greater exercise intensity. Dogs' hit rate for the lowest concentration dropped from 0.87 ± 0.04 when walking at low intensity to below 0.45 ± 0.06 when trotting at moderate intensity. This decline had an interaction with the duration of the session in moderate intensity exercise, whereby dogs performed near 100% detection in the first 10 min of the 8 km/h session, but showed 0% detection after 20 min. Hit rates for high odor concentrations (10-2) were relatively stable at both low (1 ± 0.00) and moderate (0.91 ± 0.04) exercise intensities. The paradigm and apparatus developed here may be useful to help further understand causes of operationally relevant olfactory detection threshold decline in dogs.

Development of an automated human scent olfactometer and its use to evaluate detection dog perception of human scent.

Working Dogs have shown an extraordinary ability to utilize olfaction for victim recovery efforts. Although instrumental analysis has chemically characterized odor volatiles from various human biospecimens, it remains unclear what perceptually constitutes human scent (HS) for dogs. This may be in part due to the lack of methodology and equipment to train and evaluate HS perception. The aims of this research were 1) to develop an automated human scent olfactometer (AHSO) to present HS to dogs in a controlled setting and 2) use the AHSO to evaluate dogs' response to different scented articles and individual components of HS. A human volunteer was placed in a clear acrylic chamber and using a vacuum pump and computer-controlled valves, the headspace of this chamber was carried to one of three ports in a different room. Dogs were trained to search all three ports of the olfactometer and alert to the one containing HS. In Experiment 1 and 2, the AHSO was validated by testing two dogs naïve to HS (Experiment 1) and five certified Search and Rescue (SAR) teams naïve to the apparatus (Experiment 2). All dogs showed sensitivity and specificity to HS > 95% in the apparatus. In Experiment 3, we used a spontaneous generalization paradigm to evaluate generalization from the HS chamber to different scented articles exposed to the same volunteer and to a breath sample. Dogs' response rate to the different scented articles was < 10% but exceeded 40% for the breath sample. In Experiment 4, we replicated this result by re-testing spontaneous generalization to breath and when the volunteer had breath exhausted/removed from the chamber. Dogs' response rate to breath alone was 88% and only 50% when breath was removed. Altogether, the data indicate that exhaled breath is an important and salient component of HS under these conditions.

Calibrating canines-a universal detector calibrant for detection dogs.

Since the advent of the Universal Detector Calibrant (UDC) by scientists at Florida International University in 2013, this tool has gone largely unrecognized and under-utilized by canine scent detection practitioners. The UDC is a chemical that enables reliability testing of biological and instrumental detectors. Training a biological detector, such as a scent detection canine, to respond to a safe, non-target, and uncommon compound has significant advantages. For example, if used prior to a search, the UDC provides the handler with the ability to confirm the detection dog is ready to work without placing target odor on site (i.e., a positive control), thereby increasing handler confidence in their canine and providing documentation of credibility that can withstand legal scrutiny. This review describes the UDC, summarizes its role in canine detection science, and addresses applications for UDC within scent detection canine development, training, and testing.

Evaluation of canine training aids containment for homemade explosive and components by headspace analysis and canine testing.

While canines are most commonly trained to detect traditional explosives, such as nitroaromatics and smokeless powders, homemade explosives (HMEs), such as fuel-oxidizer mixtures, are arguably a greater threat. As such, it is imperative that canines are sufficiently trained in the detection of such HMEs. The training aid delivery device (TADD) is a primary containment device that has been used to house HMEs and HME components for canine detection training purposes. This research assesses the odor release from HME components, ammonium nitrate (AN), urea nitrate (UN), and potassium chlorate (PC), housed in TADDs. Canine odor recognition tests (ORTs) were used with analytical data to determine the detectability of TADDs containing AN, UN, or PC. Headspace analysis by gas chromatography/mass spectrometry (GC/MS) with solid-phase microextraction (SPME) or online cryotrapping were used to measure ammonia or chlorine, as well as other unwanted odorants, emanating from bulk AN, UN, and PC in TADDs over 28 weeks. The analytical data showed variation in the amount of ammonia and chlorine over time, with ammonia from AN and UN decreasing slowly over time and the abundance of chlorine from PC TADDs dependent on the frequency of exposure to ambient air. Even with these variations in odor abundance, canines previously trained to detect bulk explosive HME components were able to detect all three targets in glass and plastic TADDs for at least 18 months after loading. Detection proficiency ranged from 64% to 100% and was not found to be dependent on either age of material.

The Use and Potential of Biomedical Detection Dogs During a Disease Outbreak.

Biomedical detection dogs offer incredible advantages during disease outbreaks that are presently unmatched by current technologies, however, dogs still face hurdles of implementation due to lack of inter-governmental cooperation and acceptance by the public health community. Here, we refine the definition of a biomedical detection dog, discuss the potential applications, capabilities, and limitations of biomedical detection dogs in disease outbreak scenarios, and the safety measures that must be considered before and during deployment. Finally, we provide recommendations on how to address and overcome the barriers to acceptance of biomedical detection dogs through a dedicated research and development investment in olfactory sciences.

Evaluation of a lateral flow immunoassay for the detection of the synthetic opioid fentanyl.

In 2017, 47,600 overdose deaths were reported to be associated with the abuse of opioids, including prescription painkillers (e.g. oxycodone), opiates (e.g. heroin), or synthetic opioids (e.g. fentanyl) within the United States. The recent spike in the presence of synthetic opioids in lots of heroin distributed on the street present specific and significant challenges to law enforcement. Synthetic opioids are extremely toxic substances, which can easily be inhaled. This type of exposure can lead to accidental overdoses by law enforcement and other first responders answering calls involving illicit drugs containing these substances. Due to this extreme toxicity, it is important for these individuals to have tools that can be easily deployed for accurate presumptive field tests. Currently, there are only a limited number of presumptive tests available for fentanyl detection. In this study, we addressed this technology gap by evaluating newly developed lateral flow immunoassays (LFIs) designed for the detection of fentanyl and its derivatives. These LFIs were evaluated for effectiveness in different biofluid matrices, following an in vivo exposure, cross-reactivity with fentanyl analogs, and in case samples. This study demonstrates that LFIs have the potential to be used by law enforcement for the detection of synthetic opioids.

Use of Unamplified RNA/cDNA-Hybrid Nanopore Sequencing for Rapid Detection and Characterization of RNA Viruses.

Nanopore sequencing, a novel genomics technology, has potential applications for routine biosurveillance, clinical diagnosis, and outbreak investigation of virus infections. Using rapid sequencing of unamplified RNA/cDNA hybrids, we identified Venezuelan equine encephalitis virus and Ebola virus in 3 hours from sample receipt to data acquisition, demonstrating a fieldable technique for RNA virus characterization.

Transcriptional analysis of the innate immune response of ducks to different species-of-origin low pathogenic H7 avian influenza viruses.

BACKGROUND: Wild waterfowl, including ducks, represent the classic reservoir for low pathogenicity avian influenza (LPAI) viruses and play a major role in the worldwide dissemination of AIV. AIVs belonging to the hemagglutinin (H) 7 subtype are of epidemiological and economic importance due to their potential to mutate into a highly pathogenic form of the virus. Thus far, however, relatively little work has been conducted on elucidating the host-pathogen interactions of ducks and H7 LPAIVs. In the current study, three H7 LPAIVs isolated from either chicken, duck, or turkey avian species were evaluated for their comparative effect on the transcriptional innate immune response of ducks. RESULTS: Three H7 LPAIV isolates, chicken-origin (A/chicken/Maryland/MinhMa/2004), duck-origin (A/pintail/Minnesota/423/1999), and turkey-origin (A/turkey/Virginia/SEP-67/2002) were used to infect Pekin ducks. At 3 days post-infection, RNA from spleen tissue was used for transcriptional analysis using the Avian Innate Immune Microarray (AIIM) and quantitative real-time RT-PCR (qRT-PCR). Microarray analysis revealed that a core set of 61 genes was differentially regulated in response to all three LPAIVs. Furthermore, we observed 101, 135, and 628 differentially expressed genes unique to infection with the chicken-, duck-, or turkey-origin LPAIV isolates, respectively. qRT-PCR results revealed significant (p<0.05) induction of IL-1ß, IL-2, and IFNγ transcription, with the greatest induction observed upon infection with the chicken-origin isolate. Several key innate immune pathways were activated in response to LPAIV infection including the toll-like receptor and RIG-I-like receptor pathways. CONCLUSIONS: Pekin ducks elicit a unique innate immune response to different species-of-origin H7 LPAIV isolates. However, twelve identifiable genes and their associated cell signaling pathways (RIG-I, NOD, TLR) are differentially expressed regardless of isolate origin. This core set of genes are critical to the duck immune response to AI. These data provide insight into the potential mechanisms employed by ducks to tolerate AI viral infection.

Gene expression modulation in chicken macrophages exposed to Mycoplasma synoviae or Escherichia coli.

Mycoplasma synoviae and Escherichia coli are two avian pathogens that exhibit markedly different mechanisms for infection and pathogenicity and may be expected to manipulate the host innate immune response differently. The aim of this study was to determine the extent of modulated genes and make a comparison between the transcriptomes of chicken macrophages exposed to either M. synoviae type strain WVU 1853 (MS) or avian pathogenic E. coli strain V-G (APEC). To analyze temporal gene expression profile of monocyte-derived macrophages (MDM) and HD11 cell line macrophages after each exposure, two avian immunity microarrays were used: the avian macrophage microarray (AMM) and the avian innate immunity microarray (AIIM). The quantity of MS-modulated genes was estimated in three experiments, using both microarrays. A cross-section revealed 14 AMM/AIIM genetic elements that were modulated in both types of macrophages. Additionally, to compare immunomodulatory activity of MS and APEC, MDM were exposed to each pathogen and gene modulation was detected by AIIM microarray. This study revealed 157 elements uniquely modulated by MS and 1603 elements uniquely modulated by APEC. AIIM microarray analysis also revealed a core set of 146 elements modulated by both pathogens, with generally higher induction/repression levels after APEC exposure. Validation of selected gene expression was done by quantitative real time RT-PCR. The study shows higher transcription levels of IL-1beta, IL-6, iNOS, NCF1, peroxiredoxin 1 and cathepsin L genes after MDM exposure to APEC than after exposure to MS. Surprisingly, complement component C3 gene was repressed after MDM exposure to APEC, while being induced after exposure to MS.