Single Animal Research Articles

Risk Assessment and Risk Management of Incidents of Suspected Poisoning

Abstract Poisoning in livestock, horses and wildlife may be localised or large events involving single or multiple animals and can have public health, economic or medicolegal, as well as animal welfare, implications. Investigation of these incidents is required to determine the cause, assess the risk to other animals and people and instigate an appropriate risk management plan. Investigations will involve clinical examination and history taking, site visits to determine possible sources of poisoning, post-mortem examination and collection of biological (both ante and post-mortem) and environmental samples (water, soil, feed, suspect plants). Interpretation of laboratory analyses and investigations may involve a multidisciplinary team in order to formulate and implement a risk management programme. Specialist advice should be sought early in the investigation in order to ensure appropriate sampling and correct actions are undertaken. Practitioners should be aware of where and how to report incidents, where to seek advice and have some awareness of common toxicological risks, particularly local or seasonal risks, to livestock and other animals. This case study outlines some of the steps that should be undertaken when handling a case of suspected poisoning in animals and highlights some of the possible circumstances and substances that may result in toxicosis. Information © The Author 2023

189 Single-Step Gblup Evaluation for Behavioral Traits in Labrador Retrievers Used as Guide Dogs

Abstract The objective of this study was to evaluate the accuracy of prediction in a genomic selection program for behavior traits in a population of Labrador Retrievers used as service dogs. Phenotypic data were collected in 4841 Labrador Retrievers with ages ranging from 3 months to 2.5 years, for 17 phenotypes from the International Working Dog Registry behavior checklist. The behavior checklist is a document that standardizes a scoring system for the reaction of an individual dog to environmental stimulus. Such scores are used to assess behavior and suitability of a dog for training. Pedigree contained 23,593 animals with birth dates ranging from 1991 to 2019. Genomic data were available for 457 individuals and obtained by low-pass whole genome sequences and reduced to a 250K SNP chip. Breeding values were calculated using a single trait animal model that included the fixed effects of sex, year of birth, and a contemporary group that included month/year of behavior test and organization that hosted the test. Variance components were estimated using AIREML. Genomic information was included in the model under a single-step GBLUP (ssGBLUP) approach by substituting the pedigree numerator relationship matrix with a matrix that combined pedigree and genomic relationships. Additionally, the genomic relationship matrix was modified under a weighted ssGBLUP (wssGBLUP) approach that allowed SNPs to have different distributions. Accuracies were evaluated in a 5-fold cross-validation that simulated a forward-in-time prediction. Heritabilities were low to moderate on all traits and varied from 0.025 to 0.37. Prediction based solely on pedigree information averaged 0.49 and ranged from 0.34 to 0.69. ssGBLUP increased the average accuracy to 0.55 and ranged from 0.34 to 0.72. Genomic estimated breeding values were more accurate than those computed with pedigrees for most traits. Gains in accuracy were limited by the small number of genotyped animals and are expected to increase as more animals are genotyped. The differences seen between the ssGBLUP approach and the wssGBLUP were minimal, and accuracies decreased after the second iteration. Those results indicate that behavior traits in this population are likely highly polygenic and would not benefit from weighted approaches. However, interpretation may change, as the limitations of the current study are due to the small number of genotyped individuals. Better SNP weight estimates may occur with more animals enrolled in the program, and with that a better description of the genetic architecture of the traits. The gains in accuracy show that genomic selection can help with improvement by identifying which young dogs have the highest genetic merit for the desired traits and are the best choices to keep as replacement breeders. The use of ssGBLUP is adequate for this canine data where not all animals are genotyped, and its use is recommended in selection programs focused on service dogs.

Two- and three-piece implants to boost data generation in preclinical in vivo research-A short technical report.

The purpose of this technical report is to present two novel experimental implant designs to boost data generation in preclinical in vivo research. Specifically, the report describes the rationale and the components of (1) a two-piece experimental implant suitable for a small animal platform (e.g., the rabbit femur/tibial epiphysis model), consisting of a threaded apical- and a coronal cylindrical piece, which is intended for collectingtwotypesof biomechanical data, and (2) a three-piece experimental implant suitable for a large animal platform (e.g., the mini-pig mandible model), consisting of an apical "wound chamber", which allows the collection of histological/histomorphometrical data, anda middle threadedandcoronal cylindrical piece, which also allow the collection oftwotypes of biomechanical data. The increased volume of information generated from a single experiment in a small animal platform, using the proposed two-piece implant design, may assist in a more qualified decision-making process, on whether it is relevant to proceed to further assessment using a large animal platform. Furthermore, the increased volume of information generated in a single animal experiment either in a small or large animal platform, using the proposed two- and three-piece implants, respectively, likely decreases the number of animals otherwise needed for collecting the same information with standard one-piece implants and, thus, contributes to the reduction/refinement elements of the 3R principle.

A RETROSPECTIVE ANALYSIS OF MORBIDITY AND MORTALITY IN THE CAPTIVE LEADBEATER'S POSSUM (GYMNOBELIDEUS LEADBEATERI) POPULATION FROM 1970 TO 2021.

The Leadbeater's possum (Gymnobelideus leadbeateri) is a critically endangered nocturnal marsupial with a restricted range in the Central Highlands of Victoria, Australia. There are two genetically distinct populations divided by location: highland and lowland. Lowland possums exist in one remnant swamp forest and entered captivity in 2012 when ∼60 individuals remained. Today, with less than 20 lowland individuals remaining, any information that informs the yet-unsuccessful breeding program is critical. This study encompasses a retrospective analysis of the causes of mortality and significant histological lesions in captive highland and lowland individuals across seven institutions internationally from 1970 to 2021. During this time, 245 possums lived in captivity. Postmortem records exist for 99 animals, including 349 histopathology diagnoses from 80 reports and 264 gross necropsy diagnoses from 78 reports. Diagnoses were assigned into two categories based on the importance to the individual (causing death or morbidity to a single animal [n = 194]), or importance to the wider population (causing death or morbidity to more than one animal or was related to reproduction [n = 155]). Individual animals had multiple diagnoses, which were tallied as individual data points. Renal disease was diagnosed 57 times; the most common finding was chronic nephropathy (43/57). Cardiovascular disease was diagnosed 33 times; atherosclerosis associated with obesity was common (n = 10/33). Both categories suggest causal association with captive husbandry but elicit no comment on the lack of success of the breeding program. Reproductive disease was diagnosed 36 times in 24 animals (14 females and 10 males). In females, 11 cases of uterine inflammation and associated clinical signs were associated with ascending infection or neoplasia. Of the seven lowland male possums with mortality data, five were infertile (azoospermia or testicular atrophy). More investigation into the reproductive health of this population is indicated to understand the lack of success in the current breeding program.

Milk as an indicator of dietary imbalance.

Milk provides a readily available diagnostic fluid collected daily or more frequently on an individual animal or herd basis. Milk, as an aggregated sample in bulk tank milk (BTM) represents the status of a herd instead of a single animal. In this review, we examine the potential for milk to predict risks to efficient production, reproductive success, and health on the individual cow and herd level. For many conditions related to disorders of metabolism including hyperlipdaemia and ketonaemia, improved individual cow milk testing may allow a temporally useful detection of metabolic disorder that can target intervention. However, the extension of these tests to the BTM is made more difficult by the tight temporal clustering of disorder to early lactation and the consequent mixing of cows at even moderately different stages of lactation. Integrating herd recording demographic information with Fourier-transformed mid-infrared spectra (FT-MIR) can provide tests that are useful to identify cows with metabolic disorders. The interpretation of BTM urea and protein content provides useful indications of herd nutrition. These may provide indicators that encourage further investigations of nutritional influences on herd fertility but are unlikely to provide strong diagnostic value. The fat-to-protein ratio has a high specificity, but poor sensitivity for detection of fibre insufficiency and acidosis on an individual cow basis. Selenium, zinc, β-carotene, and vitamin E status of the herd can be determined using BTM. There appears to be increasing potential for the use of milk as a diagnostic fluid as more in-parlour tests become available for individual cows. However, the BTM appears to have under-utilised potential for herd monitoring.

Spatial and phylogenetic patterns reveal hidden infection sources of Bacillus anthracis in an anthrax outbreak in Son La province, Vietnam.

Bacillus anthracis, the bacterial cause of anthrax, is a zoonosis affecting livestock and wildlife often spilling over into humans. In Vietnam, anthrax has been nationally reportable since 2015 with cases occurring annually, mostly in the northern provinces. In April 2022, an outbreak was reported in Son La province following the butchering of a water buffalo, Bubalus bubalis. A total of 137 humans from three villages were likely exposed to contaminated meat from the animal. Early epidemiological investigations suggested a single animal was involved in all exposures. Five B. anthracis isolates were recovered from human clinical cases along with one from the buffalo hide, another from associated maggots, and one from soil at the carcass site. The isolates were whole genome sequenced, allowing global, regional, and local molecular epidemiological analyses of the outbreak strains. All recovered B. anthracis belong to the A.Br.001/002 lineage based on canonical single nucleotide polymorphism analysis (canSNP). Although not previously identified in Vietnam, this lineage has been identified in the nearby countries of China, India, Indonesia, Thailand, as well as Australia. A twenty-five marker multi-locus variable number tandem repeat analysis (MLVA-25) was used to investigate the relationship between human, soil, and buffalo strains. Locally, four MLVA-25 genotypes were identified from the eight isolates. This level of genetic diversity is unusual for the limited geography and timing of cases and differs from past literature using MLVA-25. The coupled spatial and phylogenetic data suggest this outbreak originated from multiple, likely undetected, animal sources. These findings were further supported by local news reports that identified at least two additional buffalo deaths beyond the initial animal sampled in response to the human cases. Future outbreak response should include intensive surveillance for additional animal cases and additional molecular epidemiological traceback to identify pathogen sources.

The advantage of intergenic regions as genomic features for machine-learning-based host attribution of Salmonella Typhimurium from the USA.

Salmonella enterica is a taxonomically diverse pathogen with over 2600 serovars associated with a wide variety of animal hosts including humans, other mammals, birds and reptiles. Some serovars are host-specific or host-restricted and cause disease in distinct host species, while others, such as serovar S. Typhimurium (STm), are generalists and have the potential to colonize a wide variety of species. However, even within generalist serovars such as STm it is becoming clear that pathovariants exist that differ in tropism and virulence. Identifying the genetic factors underlying host specificity is complex, but the availability of thousands of genome sequences and advances in machine learning have made it possible to build specific host prediction models to aid outbreak control and predict the human pathogenic potential of isolates from animals and other reservoirs. We have advanced this area by building host-association prediction models trained on a wide range of genomic features and compared them with predictions based on nearest-neighbour phylogeny. SNPs, protein variants (PVs), antimicrobial resistance (AMR) profiles and intergenic regions (IGRs) were extracted from 3883 high-quality STm assemblies collected from humans, swine, bovine and poultry in the USA, and used to construct Random Forest (RF) machine learning models. An additional 244 recent STm assemblies from farm animals were used as a test set for further validation. The models based on PVs and IGRs had the best performance in terms of predicting the host of origin of isolates and outperformed nearest-neighbour phylogenetic host prediction as well as models based on SNPs or AMR data. However, the models did not yield reliable predictions when tested with isolates that were phylogenetically distinct from the training set. The IGR and PV models were often able to differentiate human isolates in clusters where the majority of isolates were from a single animal source. Notably, IGRs were the feature with the best performance across multiple models which may be due to IGRs acting as both a representation of their flanking genes, equivalent to PVs, while also capturing genomic regulatory variation, such as altered promoter regions. The IGR and PV models predict that ~45 % of the human infections with STm in the USA originate from bovine, ~40 % from poultry and ~14.5 % from swine, although sequences of isolates from other sources were not used for training. In summary, the research demonstrates a significant gain in accuracy for models with IGRs and PVs as features compared to SNP-based and core genome phylogeny predictions when applied within the existing population structure. This article contains data hosted by Microreact.

The draft genome of the microscopic Nemertoderma westbladi sheds light on the evolution of Acoelomorpha genomes.

Background: Xenacoelomorpha is a marine clade of microscopic worms that is an important model system for understanding the evolution of key bilaterian novelties, such as the excretory system. Nevertheless, Xenacoelomorpha genomics has been restricted to a few species that either can be cultured in the lab or are centimetres long. Thus far, no genomes are available for Nemertodermatida, one of the group's main clades and whose origin has been dated more than 400 million years ago. Methods: DNA was extracted from a single specimen and sequenced with HiFi following the PacBio Ultra-Low DNA Input protocol. After genome assembly, decontamination, and annotation, the genome quality was benchmarked using two acoel genomes and one Illumina genome as reference. The gene content of three cnidarians, three acoelomorphs, four deuterostomes, and eight protostomes was clustered in orthogroups to make inferences of gene content evolution. Finally, we focused on the genes related to the ultrafiltration excretory system to compare patterns of presence/absence and gene architecture among these clades. Results: We present the first nemertodermatid genome sequenced from a single specimen of Nemertoderma westbladi. Although genome contiguity remains challenging (N50: 60kb), it is very complete (BUSCO: 80.2%, Metazoa; 88.6%, Eukaryota) and the quality of the annotation allows fine-detail analyses of genome evolution. Acoelomorph genomes seem to be relatively conserved in terms of the percentage of repeats, number of genes, number of exons per gene and intron size. In addition, a high fraction of genes present in both protostomes and deuterostomes are absent in Acoelomorpha. Interestingly, we show that all genes related to the excretory system are present in Xenacoelomorpha except Osr, a key element in the development of these organs and whose acquisition seems to be interconnected with the origin of the specialised excretory system. Conclusion: Overall, these analyses highlight the potential of the Ultra-Low Input DNA protocol and HiFi to generate high-quality genomes from single animals, even for relatively large genomes, making it a feasible option for sequencing challenging taxa, which will be an exciting resource for comparative genomics analyses.

Structure-guided AAV capsid evolution strategies for enhanced CNS gene delivery.

Over the past 5 years, our laboratory has systematically developed a structure-guided library approach to evolve new adeno-associated virus (AAV) capsids with altered tissue tropism, higher transduction efficiency and the ability to evade pre-existing humoral immunity. Here, we provide a detailed protocol describing two distinct evolution strategies using structurally divergent AAV serotypes as templates, exemplified by improving CNS gene transfer efficiency in vivo. We outline four major components of our strategy: (i) structure-guided design of AAV capsid libraries, (ii) AAV library production, (iii) library cycling in single versus multiple animal models, followed by (iv) evaluation of lead AAV vector candidates in vivo. The protocol spans ~95 d, excluding gene expression analysis in vivo, and can vary depending on user experience, resources and experimental design. A distinguishing attribute of the current protocol is the focus on providing biomedical researchers with 3D structural information to guide evolution of precise 'hotspots' on AAV capsids. Furthermore, the protocol outlines two distinct methods for AAV library evolution consisting of adenovirus-enabled infectious cycling in a single species and noninfectious cycling in a cross-species manner. Notably, our workflow can be seamlessly merged with other RNA transcript-based library strategies and tailored for tissue-specific capsid selection. Overall, the procedures outlined herein can be adapted to expand the AAV vector toolkit for genetic manipulation of animal models and development of human gene therapies.

PERK Inhibition by HC-5404 Sensitizes Renal Cell Carcinoma Tumor Models to Antiangiogenic Tyrosine Kinase Inhibitors.

Tumors activate protein kinase R (PKR)-like endoplasmic reticulum kinase (PERK, also called EIF2AK3) in response to hypoxia and nutrient deprivation as a stress-mitigation strategy. Here, we tested the hypothesis that inhibiting PERK with HC-5404 enhances the antitumor efficacy of standard-of-care VEGF receptor tyrosine kinase inhibitors (VEGFR-TKI). HC-5404 was characterized as a potent and selective PERK inhibitor, with favorable in vivo properties. Multiple renal cell carcinoma (RCC) tumor models were then cotreated with both HC-5404 and VEGFR-TKI in vivo, measuring tumor volume across time and evaluating tumor response by protein analysis and IHC. VEGFR-TKI including axitinib, cabozantinib, lenvatinib, and sunitinib induce PERK activation in 786-O RCC xenografts. Cotreatment with HC-5404 inhibited PERK in tumors and significantly increased antitumor effects of VEGFR-TKI across multiple RCC models, resulting in tumor stasis or regression. Analysis of tumor sections revealed that HC-5404 enhanced the antiangiogenic effects of axitinib and lenvatinib by inhibiting both new vasculature and mature tumor blood vessels. Xenografts that progress on axitinib monotherapy remain sensitive to the combination treatment, resulting in ∼20% tumor regression in the combination group. When tested across a panel of 18 RCC patient-derived xenograft (PDX) models, the combination induced greater antitumor effects relative to monotherapies. In this single animal study, nine out of 18 models responded with ≥50% tumor regression from baseline in the combination group. By disrupting an adaptive stress response evoked by VEGFR-TKI, HC-5404 presents a clinical opportunity to improve the antitumor effects of well-established standard-of-care therapies in RCC.

Wide field-of-view fluorescence imaging for organ-level lineage tracing of rare intestinal stem cell populations.

Lineage tracing using fluorescent reporters is a common tool for monitoring the expression of genes and transcription factors in stem cell populations and their progeny. The zinc-binding protein 89 (ZBP-89/Zfp148 mouse gene) is a transcription factor that plays a role in gastrointestinal (GI) stem cell maintenance and cellular differentiation and has been linked to the progression of colon cancer. While lineage tracing is a useful tool, it is commonly performed with high-magnification microscopy on a small field of view within tissue sections, thereby limiting the ability to resolve reporter expression at the organ level. Furthermore, this technique requires extensive tissue processing, which is time consuming and requires euthanizing the animal. Further knowledge could be elucidated by measuring the expression of fluorescent reporters across entire organs with minimal tissue processing. We present the application of wide-field fluorescence imaging for whole-organ lineage tracing of an inducible Zfp148-tdTomato-expressing transgenic mouse line to assess the expression of ZBP-89/Zfp148 in the GI tract. We measured tdTomato fluorescence in ex vivo organs at time points between 24h and 6 months post-induction. Fluctuations in tdTomato expression were validated by fluorescence microscopy of tissue sections. Quantification of the wide field-of-view images showed a statistically significant increase in fluorescent signal across the GI tract between transgenic mice and littermate controls. The results also showed a gradient of decreasing reporter expression from proximal to distal intestine, suggesting a higher abundance of ZBP-89 expressing stem cells, or higher expression of ZBP-89 within the stem cells, in the proximal intestine. We demonstrate that wide-field fluorescence imaging is a valuable tool for monitoring whole-organ expression of fluorescent reporters. This technique could potentially be applied in vivo for longitudinal assessment of a single animal, further enhancing our ability to resolve rare stem cell lineages spatially and temporally.

Dual Tracer Test to Measure Tissue-Specific Insulin Action in Individual Mice Identifies InVivo Insulin Resistance Without Fasting Hyperinsulinemia.

The ability of metabolically active tissues to increase glucose uptake in response to insulin is critical to whole-body glucose homeostasis. This report describes the Dual Tracer Test, a robust method involving sequential retro-orbital injection of 14C-2-deoxyglucose (14C-2DG) alone, followed 40 min later by injection of 3H-2DG with a maximal dose of insulin to quantify both basal and insulin-stimulated 2DG uptake in the same mouse. The collection of both basal and insulin-stimulated measures from a single animal is imperative for generating high-quality data since differences in insulin action may be misinterpreted mechanistically if basal glucose uptake is not accounted for. The approach was validated in a classic diet-induced model of insulin resistance and a novel transgenic mouse with reduced GLUT4 expression that, despite ubiquitous peripheral insulin resistance, did not exhibit fasting hyperinsulinemia. This suggests that reduced insulin-stimulated glucose disposal is not a primary contributor to chronic hyperinsulinemia. The Dual Tracer Test offers a technically simple assay that enables the study of insulin action in many tissues simultaneously. By administering two tracers and accounting for both basal and insulin-stimulated glucose transport, this assay halves the required sample size for studies in inbred mice and demonstrates increased statistical power to detect insulin resistance, relative to other established approaches using a single tracer. The Dual Tracer Test is a valuable addition to the metabolic phenotyping toolbox.

Poster Session: Laminar and cell class distinctions for pre-saccadic attention in marmoset MT/MTC.

Pre-saccadic attention has been related to enhanced neural responses before saccades made into a neuron's receptive field in macaque visual cortex (Moore and Chang 2009). However, much remains unknown about the underlying circuit mechanisms. Using the marmoset, a small New World monkey with a smooth brain, we examined laminar and cell class distinctions during pre-saccadic attention in motion selective areas MT/MTC. In a saccade foraging task, marmosets made a saccade from a central fixation point to one of three equally eccentric random dot field stimuli. We positioned the stimuli such that one foraged location overlapped the receptive fields of neurons under study and examined how tuning functions for motion direction changed. Tuning curves were fit with an adjusted Von Mises curve that estimates baseline, gain, and tuning width. We found in two animals that neurons on average exhibited increases in baseline and gain with pre-saccadic attention, but no changes in tuning width. In a single animal we were able to dissect the population by cell class and layer. We found that increases in gain were predominantly among broad spiking neurons in superficial layers whereas additive increases in rate were shared across layers and cell types. This suggests that superficial layer broad spiking neurons, the putative projection neurons that would relay information to downstream cortical areas, have a privileged role for encoding enhanced motion sensitivity during pre-saccadic attention.

What does prey harvest composition signal to a social audience?: Experimental studies with Aché hunter-gatherers of Paraguay

In small-scale societies hunting is a high-risk, high-reward activity which impacts status and reproductive success. The question of whether men hunt to provision families or as a costly signal of their phenotypic qualities has been hotly debated in the anthropological literature. To shed new light on this question, we explored audience assessments of a hunter's phenotypic quality and desirability as a function of the composition of prey acquired by the hunter. A combination of ranking and forced-choice tasks were administered to 52 informants (46% female, aged 15–76 years) from the Aché hunter-gatherer tribe of Paraguay between May and July of 2015. Ratings of a hunter's provisioning ability, strength, fighting ability, disease resistance, and desirability as a mate or ally were all positively associated with killing large and hard-to-kill prey, and negatively associated with killing hard-to-find prey. However, killing a single large animal resulted in a worse assessment of hunter phenotype and desirability than killing an equivalent biomass of small animals. These findings highlight the potential of small prey hunting as a mechanism for advertising both quality and consistent provisioning ability. Critically, no conflict was observed between the goal of advertising quality/desirability and the goal of effective provisioning, since hunters who acquired more meat, even if the source of the meat was small game, were generally perceived as having better phenotypes and as more desirable.

The Wistar Kyoto Rat: A Model of Depression Traits.

There is an ongoing debate about the value of animal research in psychiatry with valid lines of reasoning stating the limits of individual animal models compared to human psychiatric illnesses. Human depression is not a homogenous disorder; therefore, one cannot expect a single animal model to reflect depression heterogeneity. This limited review presents arguments that the Wistar Kyoto (WKY) rats show intrinsic depression traits. The phenotypes of WKY do not completely mirror those of human depression but clearly indicate characteristics that are common with it. WKYs present despair- like behavior, passive coping with stress, comorbid anxiety, and enhanced drug use compared to other routinely used inbred or outbred strains of rats. The commonly used tests identifying these phenotypes reflect exploratory, escape-oriented, and withdrawal-like behaviors. The WKYs consistently choose withdrawal or avoidance in novel environments and freezing behaviors in response to a challenge in these tests. The physiological response to a stressful environment is exaggerated in WKYs. Selective breeding generated two WKY substrains that are nearly isogenic but show clear behavioral differences, including that of depression-like behavior. WKY and its substrains may share characteristics of subgroups of depressed individuals with social withdrawal, low energy, weight loss, sleep disturbances, and specific cognitive dysfunction. The genomes of the WKY and WKY substrains contain variations that impact the function of many genes identified in recent human genetic studies of depression. Thus, these strains of rats share characteristics of human depression at both phenotypic and genetic levels, making them a model of depression traits.

Crayfish-Tracking Motion Vision to analyze the locomotor activity induced by changes in environmental lighting

Crayfish show locomotor activity modulated by environmental lighting. Yet, little is known about different light spectra stimulation effects on the locomotor activity of crayfish, and laboratory instruments are generally limited to the study of a single animal. As such, this research describes the development and implementation of the Crayfish-Tracking Motion Vision (CTMV) system. Based on artificial vision, this system allowed for the observation and recording of locomotor activity in one to four Procambarus clarkii crayfish that were placed in individual compartments. The effect of blue or green monochromatic light stimulation was explored in two groups of dark-adapted crayfish (with a total of 15 adult crayfish). Light-emitting-diodes of blue or green spectra were applied for 5 min while locomotor activity in crayfish was monitored in a large container. Both light pulses caused similar reaction times of locomotor activity (2.9 ± 3.5 s). Meanwhile, the displacement score of “blue light” animals (2.1 ± 0.9 m) was lower than that of “green light” animals (2.8 ± 0.6 m). In contrast, velocity (1.1 ± 1 cm/s) and acceleration data (0 ± 0.8 cm/s2) of the two groups were similar. Finally, the locomotor activity decreased significantly when container size was decreased. These results showed that the CTMV system is appropriate for measuring the locomotor activities of multiple crayfish and suggested that both monochromatic lights induced different locomotor behavior in the crayfish. It has been postulated that this light-induced reflex activity, is caused by an interplay of the photoreceptor system output with a circadian system modulating light sensitivity and locomotor behavior in the crayfish,. It can be inferred that the CTMV system will benefit subsequent studies focused on light-induced reflex activity in crustaceans.

인컨텍스트 러닝을 통한 ChatGPT의 시적 표현 해석 가능성 연구

In 2023, as artificial intelligence(AI) moves beyond the realm of imitation and into the realm of creation, Korean literature is also entering the era of “creative AI,” with Slitscope’s SIA releasing a book of poetry, “시를 쓰는 이유(Why I Write Poetry).” However, the time and cost required to reach such a level is quite burdensome for the general public to attempt.
 Therefore, this study focuses on in-context learning of conversational A.I. that has already been trained through deep learning to explore the possibility of learning about A.I.’s allegory. In particular, by using ChatGPT as a research object, we can gain research advantages in terms of accessibility and functionality over previous studies on the model.
 In order to study the learning potential of ChatGPT in an in-context learning environment, this study consists of a structure of (1) pre-learning verification, (2) output request based on pre-learning, (3) related information input, and (4) verification re-execution of (2) after learning, and examines the possibility of conversational AI learning about poetic alliteration.
 The poetry texts that are input as learning information in the above process are allegorical poetry texts extracted from Yoon Gon-gang’s 『動物詩集』(Hansung Book Co., 1938) because it was judged to have value as the first Korean poetry collection with a single animal theme, which was approached and interpreted from the perspective of satirical and allegorical poetry at the time and even now.
 This study determines the learning potential of literary rhetoric for ChatGPT through the education and output process based on Yoon Gon-kang’s poems, and considers its utilization in various aspects such as education, creation, and education in the future. This will be an exploration of possibilities to solidify the position of literature in the era of artificial intelligence that will become a reality in the future.

Preclinical tendon and ligament models: Beyond the 3Rs (replacement, reduction, and refinement) to 5W1H (why, who, what, where, when, how).

Several tendon and ligament animal models were presented at the 2022 Orthopaedic Research Society Tendon Section Conference held at the University of Pennsylvania, May 5 to 7, 2022. A key objective of the breakout sessions at this meeting was to develop guidelines for the field, including for preclinical tendon and ligament animal models. This review summarizes the perspectives of experts for eight surgical small and large animal models of rotator cuff tear, flexor tendon transection, anterior cruciate ligament tear, and Achilles tendon injury using the framework: "Why, Who, What, Where, When, and How" (5W1H). A notable conclusion is that the perfect tendon model does not exist; there is no single gold standard animal model that represents the totality of tendon and ligament disease. Each model has advantages and disadvantages and should be carefully considered in light of the specific research question. There are also circumstances when an animal model is not the best approach. The wide variety of tendon and ligament pathologies necessitates choices between small and large animal models, different anatomic sites, and a range of factors associated with each model during the planning phase. Attendees agreed on some guiding principles including: providing clear justification for the model selected, providing animal model details at publication, encouraging sharing of protocols and expertise, improving training of research personnel, and considering greater collaboration with veterinarians. A clear path for translating from animal models to clinical practice was also considered as a critical next step for accelerating progress in the tendon and ligament field.

Animal models for anti-neutrophil cytoplasmic antibody-associated vasculitis: Are current models good enough?

Antineutrophil cytoplasmic autoantibody (ANCA)-associated vasculitis (AAV) is a rare and severe systemic autoimmune disease characterized by pauci-immune necrotizing inflammation of small blood vessels. AAV involves multiple organ systems throughout the body. Our knowledge of the pathogenesis of AAV has increased considerably in recent years, involving cellular, molecular and genetic factors. Because of the controlled environment with no other confounding factors, animal models are beneficial for studying the mechanistic details of disease development and for providing novel therapeutic targets with fewer toxic side effects. However, the complexity and heterogeneity of AAV make it very difficult to establish a single animal model that can fully represent the entire clinical spectrum found in patients. The aim of this review is to overview the current status of animal models for AAV, outline the pros and cons of methods, and propose potential directions for future research.

Dog owners' awareness of and motivations towards pet cadaver donation.

Animal cadavers still play an essential role in veterinary education. Many veterinary schools would like to participate in pet cadaver donation schemes for many reasons, including ethical resourcing of cadavers, but finding owners willing to donate their pet is challenging. The aim of this study was to investigate dog owners' awareness of pet cadaver donation. The study was conducted using Jisc online surveys with the survey being distributed via social media. In total, 708 people answered a 10-question online questionnaire. The most common reason given for why a dog owner might donate was to assist in the education of future veterinarians (n = 396, 56%). When asked to rank the options given to them upon the passing of their pet, single animal cremation was overwhelmingly the participants' favoured option (n = 483, 68.2%). Factors that might affect the findings, such as religion, financial status and cultural differences, were not explored in this study. The only incentive shown to increase the likelihood of pet owners donating to a pet cadaver scheme was the return their pet's ashes to them after the cadaver had served its purpose. This study revealed that dog owners currently have very little knowledge of pet cadaver donation.