Using time-lapse digital video recording for a nesting study of birds of prey

Remote photography using various photo, movie or video devices has been employed in numerous studies in wildlife research during the last 50 years. Given the rapid advances in digital technologies, digital video and photo techniques are becoming more common in use, and publications that introduce a new method or equipment for video surveillance in wildlife research (and in ornithological studies particularly) are appearing almost every year. However, still no special guide to the great variety of equipment and methods is available, and the choice and use of suitable gear for scientific purposes may be difficult for non-specialists. In this paper, we review the most common surveillance techniques used in today’s nest studies, as well as the most essential properties of image recording equipment. We also describe the digital video recording technique, which we used for observations of raptor nests, and summarise our experience of its operation. As an example of the obtained data, we present the timing of prey deliveries of goshawks and common buzzards.     

Construction of a thermotaxis chamber providing spatial or temporal thermal gradients monitored by an infrared video camera system.

A thermotaxis chamber was constructed to quantitatively study thermotaxis in eukaryotic amoeboid cells. The apparatus provided either spatial or temporal temperature gradients in an observation chamber set in an inverted microscope. With an infrared video camera system, spatial thermal gradients were monitored directly and the temperature at the actual location of the cells could be estimated accurately. This enabled a precise determination of the strength of thermal stimuli. With this apparatus, we were able to simultaneously measure temperature and observe cellular behavior directly. This feature permits quantitative studies on stimulus-response relationships. The utility of the apparatus was demonstrated by thermotaxis assay under a spatial thermal gradient in polymorphonuclear leukocytes. Since this apparatus can also provide temporal thermal gradients, it may have several applications in studies of temperature-dependent phenomena in cell biology.     

A microcomputer interface for a digital audio processor-based data recording system.

An inexpensive interface is described that performs direct transfer of digitized data from the digital audio processor and video cassette recorder based data acquisition system designed by Bezanilla (1985, Biophys. J., 47:437-441) to an IBM PC/XT microcomputer. The FORTRAN callable software that drives this interface is capable of controlling the video cassette recorder and starting data collection immediately after recognition of a segment of previously collected data. This permits piecewise analysis of long intervals of data that would otherwise exceed the memory capability of the microcomputer.     

A high capacity data recording device based on a digital audio processor and a video cassette recorder.

A modified digital audio processor, a video cassette recorder, and some simple added circuitry are assembled into a recording device of high capacity. The unit converts two analog channels into digital form at 44-kHz sampling rate and stores the information in digital form in a common video cassette. Bandwidth of each channel is from direct current to approximately 20 kHz and the dynamic range is close to 90 dB. The total storage capacity in a 3-h video cassette is 2 Gbytes. The information can be retrieved in analog or digital form.     

Applications of video mixing and digital overlay to neuroethology

Neuroethological experiments often require video images of animal behavior and recordings of physiological data to be acquired simultaneously, synchronized with each other, stored, and analyzed together. The use of inexpensive multimedia computers offers new possibilities for mixing video images, analog voltages, and computer data, storing these combined signals to videotape, and extracting quantitative data for analysis. In this paper, we summarize methods for mixing images from multiple video cameras and a Macintosh computer display to facilitate manipulation of data generated during our neurophysiological and behavioral research. These technologies enhance accuracy, speed, and flexibility during experiments, and facilitate selecting and extracting quantitative data from the videotape for further analysis. Three applications are presented: (A) we used an analog video mixer to synchronize neurophysiological recordings with ongoing behaviors of freely moving rats; (B) we used a chroma keyed digital overlay to generate positional data for the rat's face during drinking behavior; and (C) we combined a computer model of a rat's head and whiskers with videos of exploratory behaviors to better track and quantify movements in three dimensions. Although the applications described here are specific to our neuroethological work, these methods will be useful to anyone wishing to combine the signals from multiple video sources into a single image or to extract series of positional or movement data from video frames without frame grabbing.     

Low-cost conversion of the Polaroid MD-4 land camera to a digital gel documentation system

A simple, inexpensive design is presented for the rapid conversion of the popular MD-4 Polaroid land camera to a high quality digital gel documentation system. Images of ethidium bromide stained DNA gels captured using the digital system were compared to images captured on Polaroid instant film. Resolution and sensitivity were enhanced using the digital system. In addition to the low cost and superior image quality of the digital system, there is also the added convenience of real-time image viewing through the swivel LCD of the digital camera, wide flexibility of gel sizes, accurate automatic focusing, variable image resolution, and consistent ease of use and quality. Images can be directly imported to a computer by using the USB port on the digital camera, further enhancing the potential of the digital system for documentation, analysis, and archiving. The system is appropriate for use as a start-up gel documentation system and for routine gel analysis.     

Development of a versatile computer integrated control system for bioprocess controls

A general approach is described for the implementation of a networked multi-unit computer integrated control system. The use of data acquisition hardware and graphical programming tools alleviates tedious programming and maintains potency and flexibility. One application of the control system, the control of a mammalian cell perfusion culture based on a key nutrient glucose concentration, was demonstrated. The control system offers customized user interface for all process control parameters and allows the flexibility for continued improvement and implementation of new tailored functions. The temperature, pH, dissolved oxygen and glucose level were accurately controlled. This revised version was published online in June 2006 with corrections to the Cover Date.     

Development of a room laser based real-time alignment monitoring system using an array of photodiodes

PurposeTo develop a real-time alignment monitoring system (RAMS) to compensate for the limitations of the conventional room-laser-based alignment system. To verify the feasibility of the RAMS, reproducibility and accuracy tests were conducted.MethodsRAMS was composed of a room laser sensing array (RLSA), an electric circuit, an analog-to-digital converter (ADC), and a control PC. The RLSA was designed to arrange photodiodes in a pattern that results in the RAMS having a resolution of 1 mm. The photodiodes were used for quantitative assessment of the alignment condition. To verify the usability of the developed system, we conducted tests of temporal reproducibility, repeatability, and accuracy.ResultsThe results of the temporal reproducibility test suggested that the signal of the RAMS was stable with respect to time. Further, the repeatability test resulted in a maximum coefficient of variance of 1.14%, suggesting that the signal of the RAMS was stable over repeated set-ups. The accuracy test confirmed that the “on” and “off” signals could be distinguished by signal intensity, considering that the “off” signal was below 75% of the “on” signal in every case. In addition, we confirmed that the system can detect 1 mm of movement by monitoring the pattern of the “on” and “off” signals.ConclusionWe developed a room laser based alignment monitoring system. The feasibility test verified that the system is capable of quantitative alignment monitoring in real time. We expect that the RAMS can propose the potential of the room laser based alignment monitoring method.     

猪圆环病毒2型TaqMan实时PCR检测方法的建立

设计合成了一套引物和TaqMan探针,特异性扩增猪圆环病毒2型（PCV2）ORF2基因,在国内首次建立了快速定量检测PCV2的实时PCR方法,且该方法具有较好的特异性和重复性,对PCV2DNA检测下限为1copy／μL,敏感性比常规PCR高10^6倍;分别用该法和普通PCR方法对PMWS人工发病猪的10份组织及30份血清样品检测,结果表明该方法具有更快速、灵敏、准确、低污染等优点,并可以对PMWS的早期检测、预防起到指示作用。     

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.     

The Strep-tag system for one-step purification and high-affinity detection or capturing of proteins

The Strep-tag II is an eight-residue minimal peptide sequence (Trp-Ser-His-Pro-Gln-Phe-Glu-Lys) that exhibits intrinsic affinity toward streptavidin and can be fused to recombinant proteins in various fashions. We describe a protocol that enables quick and mild purification of corresponding Strep-tag II fusion proteins--including their complexes with interacting partners--both from bacterial and eukaryotic cell lysates using affinity chromatography on a matrix carrying an engineered streptavidin (Strep-Tactin), which can be accomplished within 1 h. A high-affinity monoclonal antibody (StrepMAB-Immo) permits stable immobilization of Strep-tag II fusion proteins to solid surfaces, for example, for surface plasmon resonance analysis. Selective and sensitive detection on western blots is achieved with Strep-Tactin/enzyme conjugates or another monoclonal antibody (StrepMAB-Classic). Thus, the Strep-tag II, which is short, biologically inert, proteolytically stable and does not interfere with membrane translocation or protein folding, offers a versatile tool both for the rapid isolation of a functional gene product and for its detection or molecular interaction analysis.     

Development of a TaqMan quantitative PCR assay specific for Cryptosporidium parvum

A rapid detection method that is both quantitative and specific for the water-borne human parasite Cryptosporidium parvum is reported. Real-time polymerase chain reaction (PCR) combined with fluorescent TaqMan technology was used to develop this sensitive and accurate assay. The selected primer-probe set identified a 138-bp section specific to a C. parvum genomic DNA sequence. The method was optimized on a cloned section of the target DNA sequence, then evaluated on C. parvum oocyst dilutions. Quantification was accomplished by comparing the fluorescence signals obtained from test samples of C. parvum oocysts with those obtained from standard dilutions of C. parvum oocysts. This real-time PCR assay allowed reliable quantification of C. parvum oocysts over six orders of magnitude with a baseline sensitivity of six oocysts in 2 h.     

Development of a rotor-gene real-time PCR assay for the detection and quantification of Mycoplasma genitalium

We developed and validated a real-time quantitative polymerase chain reaction (qPCR) assay to determine Mycoplasma genitalium bacterial load in endocervical swabs, based on amplification of the pdhD gene which encodes dihydrolipoamide dehydrogenase, using the Rotor-Gene platform. We first determined the qPCR assay sensitivity, limit of detection, reproducibility and specificity, and then determined the ability of the qPCR assay to quantify M. genitalium in stored endocervical specimens collected from Zimbabwean women participating in clinical research undertaken between 1999 and 2007. The qPCR assay had a detection limit of 300 genome copies/mL and demonstrated low intra- and inter-assay variability. The assay was specific for M. genitalium DNA and did not amplify the DNA from other mycoplasma and ureaplasma species. We quantified M. genitalium in 119 of 1600 endocervical swabs that tested positive for M. genitalium using the commercial Sacace M. genitalium real-time PCR, as well as 156 randomly selected swabs that were negative for M. genitalium by the same assay. The M. genitalium loads ranged between < 300 and 3,240,000 copies/mL. Overall, the qPCR assay demonstrated good range of detection, reproducibility and specificity and can be used for both qualitative and quantitative analyses of M. genitalium in endocervical specimens and potentially other genital specimens.     

猪繁殖与呼吸综合征病毒实时PCR检测方法的建立

设计合成了一套引物和TaqMan探针,以扩增猪繁殖与呼吸综合征病毒的核衣壳蛋白基因,通过反应条件的优化,在国内首次建立了快速定量检测猪繁殖与呼吸综合征病毒的实时PCR方法,并用该法检测患病猪的肺脏等样品。结果表明该方法具有较好的特异性和重复性,对PRRSV细胞培养物的检测下限为0.01TCID50,敏感性比常规RT-PCR高100倍;对10份PRRS疑似猪肺脏样品检测5份为阳性,与病毒分离的阳性符合率为100%。该方法具有快速、灵敏、准确、低污染等优点,在PRRSV的早期检测、预防控制、进出口检疫及基础研究中会起到重要作用。     

DScan - a high-performance digital scanning system for entomological collections

Here we describe a high-performance imaging system for creating high-resolution images of whole insect drawers. All components of the system are industrial standard and can be adapted to meet the specific needs of entomological collections. A controlling unit allows the setting of imaging area (drawer size), step distance between individual images, number of images, image resolution, and shooting sequence order through a set of parameters. The system is highly configurable and can be used with a wide range of different optical hardware and image processing software.     

Performance of a digital video camcorder for the Autonomous Biological System experiment onboard Space Station Mir.

A video imaging and recording system was utilized in the Autonomous Biological System experiment onboard the space station Mir. Video image of the mini-ecological system was successfully recorded. The whole system was retrieved to the ground after its operation in orbit for four months. Performance of the video system is summarized here together with technical problems encountered. Defects of pixel had been developed in the imaging device. Cause of these defects could be attributed to its exposure against space radiation. Auto white balance was another function of the camcorder that was deviated from normal range of its performance once in orbit but recovered to normal after a while. Possible use of imaging devices for dosimetry is proposed to record space radiation environment at the site close to the experiment.