动物离心机中大鼠行为状态无线视频采集装置的研制

目的：研制一种在正加速度作用下能实时观测大鼠行为状态的实验视频采集装置,为研究正加速度导致脑损伤的病理生理机制和防治措施提供新的方法。方法：依据正加速度对大鼠体位的要求和离心机特殊的环境,通过特制的大鼠座舱和无线视频采集单元来实现对动物离心机中正加速度作用下大鼠行为状态的实时观测;固定大鼠的座舱主要由胸部固定圈、身长调节圈、头部运动限制框构成;无线视频采集单元由2.4G无线摄像头、无线信号接收器、信号放大器、计算机以及视频采集软件组成。结果：座舱特殊的大鼠固定方式,既满足了正加速度对体位的要求,又不影响对大鼠行为状态的观测,2．4G无线视频采集技术保证视频信号在离心机高速旋转环境下的实时传输和观测,视频画面清晰流畅,并且能保存视频供回放分析。结论：动物离心机无线视频采集装置可用于正加速度作用下大鼠行为状态的实时观测与研究。     

BEEtag: A Low-Cost,Image-Based Tracking System for the Study of Animal Behavior and Locomotion

A fundamental challenge common to studies of animal movement, behavior, and ecology is the collection of high-quality datasets on spatial positions of animals as they change through space and time. Recent innovations in tracking technology have allowed researchers to collect large and highly accurate datasets on animal spatiotemporal position while vastly decreasing the time and cost of collecting such data. One technique that is of particular relevance to the study of behavioral ecology involves tracking visual tags that can be uniquely identified in separate images or movie frames. These tags can be located within images that are visually complex, making them particularly well suited for longitudinal studies of animal behavior and movement in naturalistic environments. While several software packages have been developed that use computer vision to identify visual tags, these software packages are either (a) not optimized for identification of single tags, which is generally of the most interest for biologists, or (b) suffer from licensing issues, and therefore their use in the study of animal behavior has been limited. Here, we present BEEtag, an open-source, image-based tracking system in Matlab that allows for unique identification of individual animals or anatomical markers. The primary advantages of this system are that it (a) independently identifies animals or marked points in each frame of a video, limiting error propagation, (b) performs well in images with complex backgrounds, and (c) is low-cost. To validate the use of this tracking system in animal behavior, we mark and track individual bumblebees (Bombus impatiens) and recover individual patterns of space use and activity within the nest. Finally, we discuss the advantages and limitations of this software package and its application to the study of animal movement, behavior, and ecology.     

BirdOriTrack: a new video‐tracking program for orientation research with migratory birds

Experimental research on the orientation of migratory songbirds is traditionally conducted using orientation funnels or automatic registration cages that record the directional activity of individual birds on paper or a computer. Most traditionally used funnel and cage designs do not permit investigators to observe detailed behavior of the birds and, therefore, we have gained little insight into the actual behavior of birds while they are exhibiting migratory restlessness and making directional choices. Such behavior can only be studied by direct observation or by video filming. Here, we present BirdOriTrack, a video‐tracking program for extracting time‐resolved, positional data of birds (and potentially other animal species) to determine their orientation relative to the center of a circular cage/funnel. With relatively inexpensive cameras, recording equipment, and cages, orientation experiments can easily be conducted and analyzed using BirdOriTrack. The program is designed to be flexible, allowing analysis of orientation behavior of birds of any size in different cage designs and in both controlled laboratory settings and field‐based studies. To demonstrate the program's utility, we show the results of preliminary field experiments on several species of migratory birds captured at a migration monitoring station. BirdOriTrack is freely available at http://canmove.lu.se/birdoritrack .     

A novel acoustic evaluation system of scratching in mouse dermatitis: rapid and specific detection of invisibly rapid scratch in an atopic dermatitis model mouse

Scratching is an essential and a skin specific behavior induced by itching, which is a common symptom of atopic dermatitis (AD) and other types of dermatitis. Itching sensation and scratching are closely associated and thus scratching times are currently used for evaluating itching in animal models. However, objective measuring systems of scratching to assess the grade of dermatitis and the effectiveness of anti-pruritic drugs in animal dermatitis models are lacking. To investigate a quantitative evaluation system for itching, we have developed a novel acoustic scratching counting system and compared its accuracy with time-consuming slow-motion video recording system. We have also objectively evaluated the efficacy of an antihistamine using this novel system. Scratching behavior of an AD model mouse (K14 driven IL-18 transgenic mouse) was recorded visually and acoustically. Specific scratching sound produced by mice was recorded and counted using a software we have developed, and the results obtained using our acoustic system were not statistically different from data obtained using slow motion video system. Surprisingly, mice scratched more than 10 times/second, which was invisibly rapid motion and revealed inaccuracy of conventional hand counting system. Results were identical to that of measured by 10 times time costing slow-motion video analysis. The antihistamine is clearly effective for suppression of scratching as demonstrated using this objective and accurate method. This novel motion analysis system will open a window for physiological and pathological analysis for animal models and development of anti-pruritic drugs.     

Meal Duration as a Measure of Orofacial Nociceptive Responses in Rodents

A lengthening in meal duration can be used to measure an increase in orofacial mechanical hyperalgesia having similarities to the guarding behavior of humans with orofacial pain. To measure meal duration unrestrained rats are continuously kept in sound attenuated, computerized feeding modules for days to weeks to record feeding behavior. These sound-attenuated chambers are equipped with chow pellet dispensers. The dispenser has a pellet trough with a photobeam placed at the bottom of the trough and when a rodent removes a pellet from the feeder trough this beam is no longer blocked, signaling the computer to drop another pellet. The computer records the date and time when the pellets were taken from the trough and from this data the experimenter can calculate the meal parameters. When calculating meal parameters a meal was defined based on previous work and was set at 10 min (in other words when the animal does not eat for 10 min that would be the end of the animal''s meal) also the minimum meal size was set at 3 pellets. The meal duration, meal number, food intake, meal size and inter-meal interval can then be calculated by the software for any time period that the operator desires. Of the feeding parameters that can be calculated meal duration has been shown to be a continuous noninvasive biological marker of orofacial nociception in male rats and mice and female rats. Meal duration measurements are quantitative, require no training or animal manipulation, require cortical participation, and do not compete with other experimentally induced behaviors. These factors distinguish this assay from other operant or reflex methods for recording orofacial nociception.     

Long-term Behavioral Tracking of Freely Swimming Weakly Electric Fish

Long-term behavioral tracking can capture and quantify natural animal behaviors, including those occurring infrequently. Behaviors such as exploration and social interactions can be best studied by observing unrestrained, freely behaving animals. Weakly electric fish (WEF) display readily observable exploratory and social behaviors by emitting electric organ discharge (EOD). Here, we describe three effective techniques to synchronously measure the EOD, body position, and posture of a free-swimming WEF for an extended period of time. First, we describe the construction of an experimental tank inside of an isolation chamber designed to block external sources of sensory stimuli such as light, sound, and vibration. The aquarium was partitioned to accommodate four test specimens, and automated gates remotely control the animals'' access to the central arena. Second, we describe a precise and reliable real-time EOD timing measurement method from freely swimming WEF. Signal distortions caused by the animal''s body movements are corrected by spatial averaging and temporal processing stages. Third, we describe an underwater near-infrared imaging setup to observe unperturbed nocturnal animal behaviors. Infrared light pulses were used to synchronize the timing between the video and the physiological signal over a long recording duration. Our automated tracking software measures the animal''s body position and posture reliably in an aquatic scene. In combination, these techniques enable long term observation of spontaneous behavior of freely swimming weakly electric fish in a reliable and precise manner. We believe our method can be similarly applied to the study of other aquatic animals by relating their physiological signals with exploratory or social behaviors.     

Long-term automatic video recording as a tool for analysing the time patterns of utilisation of predefined locations by wild animals

The design and application of automatic video recording systems for wild animals are described. Such systems enable continuous, long-time and repercussion-free surveillance of selected areas in the field. The performance characteristics of a conventional VHS video-tape system are compared to a digital video recording system. The recordings were used to develop daily and annual plots of occurrence for the different species and to display the preferences for darkness, twilight and light phase by the different species over the year. Absolute utilisation frequency as well as relative species composition can be obtained and compared between seasons or different places. The videos also were analysed for time budgets of basic behaviour patterns like feeding, walking, observing, social interaction, flight and comfort behaviour. Automatic video technique is a highly convenient tool for systematic long-term field research on occurrence of wild animal species, daily and annual activity rhythms, behaviour and area utilisation. Such systems can be applied especially to record animals at clearings, feeding or bait places, water sources, salt licks or traps. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

川金丝猴射精标准的量化辅助

通过对位于秦岭周至国家级自然保护区玉皇庙西梁川金丝猴群的交配行为进行拍摄记录(2003 年7～12 月), 运用NOLDUS公司编制的The Observer 610 动物行为分析系统对摄像资料进行分析, 发现野生川金丝猴采用腹背式交配方式, 交配中(N = 285) 有单足踩脚踝(54157%) 和双足踩脚踝(45143%) 两种姿势, 与圈养观察中仅发现单足踩脚踝交配姿势有别。此外, 通过与笼养和圈养条件下报道的射精标准比较, 我们将抽插过程分为“抽插前期”和“抽插后期”两个阶段, 并试图为川金丝猴交配过程中的射精行为判定提供一个量化辅助。     

Time-lapse infrared videography for animal behavior observations

Although videography has been used for many years to record episodes of animal behavior, the availability of increasingly complex equipment can make selection difficult. Most equipment developed for round-the-clock observations was designed for security surveillance, so informed purchasing, installation, and usage decisions need to be made to monitor animal behavior effectively. During 7 years of using video and infrared equipment for animal behavior research and monitoring in a zoo setting, we tested many models, evaluated options, and modified components. We now have well-integrated systems that are compatible with our video/computer review and analysis program. New equipment will continue to become available, but familiarity with basic features and applications can help to narrow the selection. Zoo Biol 17:535–543, 1998. © 1998 Wiley-Liss, Inc.     

Bee Tracker—an open‐source machine learning‐based video analysis software for the assessment of nesting and foraging performance of cavity‐nesting solitary bees

The foraging and nesting performance of bees can provide important information on bee health and is of interest for risk and impact assessment of environmental stressors. While radiofrequency identification (RFID) technology is an efficient tool increasingly used for the collection of behavioral data in social bee species such as honeybees, behavioral studies on solitary bees still largely depend on direct observations, which is very time‐consuming. Here, we present a novel automated methodological approach of individually and simultaneously tracking and analyzing foraging and nesting behavior of numerous cavity‐nesting solitary bees. The approach consists of monitoring nesting units by video recording and automated analysis of videos by machine learning‐based software. This Bee Tracker software consists of four trained deep learning networks to detect bees that enter or leave their nest and to recognize individual IDs on the bees’ thorax and the IDs of their nests according to their positions in the nesting unit. The software is able to identify each nest of each individual nesting bee, which permits to measure individual‐based measures of reproductive success. Moreover, the software quantifies the number of cavities a female enters until it finds its nest as a proxy of nest recognition, and it provides information on the number and duration of foraging trips. By training the software on 8 videos recording 24 nesting females per video, the software achieved a precision of 96% correct measurements of these parameters. The software could be adapted to various experimental setups by training it according to a set of videos. The presented method allows to efficiently collect large amounts of data on cavity‐nesting solitary bee species and represents a promising new tool for the monitoring and assessment of behavior and reproductive success under laboratory, semi‐field, and field conditions.     

Formation of behavior in bottle-nosed dolphin newborns under conditions of an oceanarium: Heterochrony of involvement of various types of afferentiation

The formation of behavior in early ontogenesis in dolphin newborns kept in sections of marine enclosures was studied using direct observations and photo- and video surveys, as well as by recording ethograms with subsequent computer analysis. A successive formation of postures and locomotory acts accompanied by a heterochronous involvement of various types of afferentiation was found.     

Comparison of video recording and a portable electronic device for measuring the feeding behaviour of individually housed dairy goats

A detailed knowledge of chewing behaviour is important in understanding the factors that can affect digestive function in high producing ruminants. The aim of this study was to compare chewing behaviour either measured with a portable automatic system (APEC) which records jaw movements or obtained by scan sampling video analysis in 12 individually housed dairy goats. One hour and daily time-intervals were used for the analysis. APEC and video showed better agreement for 1 h than for daily time-intervals but a very high individual variability was observed for all the parameters measured. Daily duration of rumination seemed to be assessed better by the APEC than by the video because it is sometimes difficult to determine if the goats are resting or ruminating on the video. Daily duration of intake was however assessed better by the video because the automatic recorder interpreted all oral behaviours as intake. However, total chewing time is directly related to the amount of saliva produced which an important factor in ruminant nutrition. The APEC allows continuous recording of activity independently of the animal's position in the pen or in a group or at pasture where animals cannot be filmed, however the device can be easily damaged by the animals. Video cannot be damaged by the animals but it can be difficult to determine the animal's activity if the animal is lying-down or not facing the camera, or in a dark part of its pen. Both systems were relevant to measure chewing behaviour of stall housed dairy goats, and to study the daily time-budget of feeding behaviour but results have to be interpreted carefully.     

Bring your own camera to the trap: An inexpensive,versatile, and portable triggering system tested on wild hummingbirds

The study of animals in the wild offers opportunities to collect relevant information on their natural behavior and abilities to perform ecologically relevant tasks. However, it also poses challenges such as accounting for observer effects, human sensory limitations, and the time intensiveness of this type of research. To meet these challenges, field biologists have deployed camera traps to remotely record animal behavior in the wild. Despite their ubiquity in research, many commercial camera traps have limitations, and the species and behavior of interest may present unique challenges. For example, no camera traps support high‐speed video recording. We present a new and inexpensive camera trap system that increases versatility by separating the camera from the triggering mechanism. Our system design can pair with virtually any camera and allows for independent positioning of a variety of sensors, all while being low‐cost, lightweight, weatherproof, and energy efficient. By using our specialized trigger and customized sensor configurations, many limitations of commercial camera traps can be overcome. We use this system to study hummingbird feeding behavior using high‐speed video cameras to capture fast movements and multiple sensors placed away from the camera to detect small body sizes. While designed for hummingbirds, our application can be extended to any system where specialized camera or sensor features are required, or commercial camera traps are cost‐prohibitive, allowing camera trap use in more research avenues and by more researchers.     

VBA开发动物行为实时记录系统及实例

对动物行为进行实时记录是行为学研究的一项重要工作。传统的行为观察方法常常无法满足实时精确记录行为的要求 ,而一些现代的行为记录系统虽功能强大但却难以推广。作者应用VBA开发了一个简捷方便的行为实时记录系统 ,可以减少记录动物行为观察的工作量 ,降低记录动物行为的观察误差 ,保证了实验数据的可靠性。同时 ,该系统具有开放性和灵活性的特点 ,可以在不同动物行为研究中应用。     

Video Bioinformatics Analysis of Human Embryonic Stem Cell Colony Growth

Because video data are complex and are comprised of many images, mining information from video material is difficult to do without the aid of computer software. Video bioinformatics is a powerful quantitative approach for extracting spatio-temporal data from video images using computer software to perform dating mining and analysis. In this article, we introduce a video bioinformatics method for quantifying the growth of human embryonic stem cells (hESC) by analyzing time-lapse videos collected in a Nikon BioStation CT incubator equipped with a camera for video imaging. In our experiments, hESC colonies that were attached to Matrigel were filmed for 48 hours in the BioStation CT. To determine the rate of growth of these colonies, recipes were developed using CL-Quant software which enables users to extract various types of data from video images. To accurately evaluate colony growth, three recipes were created. The first segmented the image into the colony and background, the second enhanced the image to define colonies throughout the video sequence accurately, and the third measured the number of pixels in the colony over time. The three recipes were run in sequence on video data collected in a BioStation CT to analyze the rate of growth of individual hESC colonies over 48 hours. To verify the truthfulness of the CL-Quant recipes, the same data were analyzed manually using Adobe Photoshop software. When the data obtained using the CL-Quant recipes and Photoshop were compared, results were virtually identical, indicating the CL-Quant recipes were truthful. The method described here could be applied to any video data to measure growth rates of hESC or other cells that grow in colonies. In addition, other video bioinformatics recipes can be developed in the future for other cell processes such as migration, apoptosis, and cell adhesion. Download video file.^{(111M, mp4)}     

数码显微镜DIY及应用探讨

数码显微镜相对于光学显微镜而言,在观察效果、指示效果、图像记录、实验效率等方面均有很大的优点。作者利用现有的光学显微镜和一般的电脑摄像头,经过组装与调试,DIY成了完全可以实用的数码显微镜。并提供了作者在实际应用中总结的一些经验：如硬件和软件的选择与使用,图像的显示、指示、捕获、存储及动态摄像等。     

Particle detection,number estimation,and feature measurement in gene transfer studies: optical fractionator stereology integrated with digital image processing and analysis

Assessing the efficacy of in vivo gene transfer often requires a quantitative determination of the number, size, shape, or histological visualization characteristics of biological objects. The optical fractionator has become a choice stereological method for estimating the number of objects, such as neurons, in a structure, such as a brain subregion. Digital image processing and analytic methods can increase detection sensitivity and quantify structural and/or spectral features located in histological specimens. We describe a hardware and software system that we have developed for conducting the optical fractionator process. A microscope equipped with a video camera and motorized stage and focus controls is interfaced with a desktop computer. The computer contains a combination live video/computer graphics adapter with a video frame grabber and controls the stage, focus, and video via a commercial imaging software package. Specialized macro programs have been constructed with this software to execute command sequences requisite to the optical fractionator method: defining regions of interest, positioning specimens in a systematic uniform random manner, and stepping through known volumes of tissue for interactive object identification (optical dissectors). The system affords the flexibility to work with count regions that exceed the microscope image field size at low magnifications and to adjust the parameters of the fractionator sampling to best match the demands of particular specimens and object types. Digital image processing can be used to facilitate object detection and identification, and objects that meet criteria for counting can be analyzed for a variety of morphometric and optical properties.     

Quick and efficient measurement of transient GUS expression using image analysis

An image-analysis procedure was developed for measurement of transient β-glucuronidase (GUS) gene expression events in peanut. Gene transfer via microprojectile bombardment has been effective in peanut. Experiments were performed to improve the efficiency of DNA delivery to embryogenic calli for transformation. Transient GUS expression was assayed 24 hours after bombardment by staining calli and manually counting blue-stained spots under a microscope. These calli were then placed under a dissecting microscope with a video camera that recorded each image onto a computer. The images were then subjected to analysis with software that counted the number of blue-stained GUS-positive spots as well as the area and percent of the total area of each spot. The manual counts and data from the image analysis were compared through correlation statistics and by comparing analyses of variance for fixed parameters of the experiments. Correlation coefficients were high between manual spot counts and image-analysis counts (r=0.77) and between manual counts and percent area (GUS (r=0.91) Results indicate that image analysis provides a quick and efficient method for analysis of transient expression. Much more reliable information is obtained with less effort and less fatigue with the image-analysis procedure. Scientists can use this method to monitor transient expression events in plant transformation experiments.     

Regulatory Considerations for the Classification of Video Monitoring in Dissolution Testing

The goal of this article is to discuss the classification of video recordings and images when applied to dissolution testing in USP apparatus 1 and 2. Three use cases are presented. The first case presents the use and classification of video as RBE (review by exception) data under GAMP 5. The second case presents the use of video in formulation development in a research and development environment. The third case presents a feasibility study using readily available computer vision software to recognize and measure objects in the dissolution vessel, setting the groundwork for the use of image analysis as a quantitative tool. The classification of video as “electronic data”, requiring 21 CFR part 11 compliance, versus its classification as a RBE data under GAMP 5, likely depends upon its use case. Another goal of this article is to establish a position on the use of video monitoring technology as a tool for dissolution testing that is fit for purpose and compliant with regulations regarding video data management and information.Key words: computer vision, dissolution, GAMP, USP, video monitoring