Time-lapse Video Recording Research Articles

Sitting Behavior in Lactating Sows was not Related to Longevity and Lifetime Performance

The stereotypy behavior is defined as the repetitive and relatively invariant behavior pattern with no obvious function. Increased stereotypy behavior raises a concern for poor well-being for animals. The dog-sitting (sitting) behavior in sows is considered as one of the stereotypies. The objectives of this study were to investigate the relationships between sitting behaviors and longevity of sows and to compare sitting behaviors among sows culled for “locomotion” and “other reasons.” A commercial farrowing-to-finish farm with approximately 300 female pigs was visited 11 times for 3 years to record the behaviors of sows using time-lapse video recorders for 24hr. Sitting behaviors were measured as the total duration of sitting postures for 24hr and the average duration of sitting bouts The lifetime records of each sow were collected using a software system. Sows were categorized into two groups of : high and low longevity. High longevity groups included the sows having both parity at cullin≥6 and the culling reason was “old age.” The other sows were categorized into the low longevity groups. Sows culled at the observed parity were divided into two groups on the basis of culling reasons : sows culled for “locomotion” and sows culled for “other reasons.” Among the 205 sows observed, means of total duration of sitting postures for 24hr and average duration of sitting bouts were found to be 80.7±3.44min and 2.6±0.11min, respectively. No differences were found in the total duration of sitting postures and average duration of sitting bouts between the two longevity groups. Additionally, both sitting behaviors were not related with herd-entry to culling interval and lifetime pigs born alive. The groups, which were classified on the basis of culling reasons, showed no differences in terms of sitting behaviors. In conclusion, sitting behaviors in lactating sows were not related to longevity and lifetime performance.

Improvement of Flower Vase Life using Cross-Breeding Techniques in Carnation (Dianthus caryophyllus L.)

The vase life of cut flowers, or flower longevity, is one of the most important characteristics that deter- mine their quality and their ability to satisfy consumer preferences. To improve the vase life of carna- tions, conventional cross-breeding techniques were used to develop many carnation lines with a long vase life. Two new cultivars, 'Miracle Rouge' and 'Miracle Symphony', with a genetically determined long vase life, were developed. They had vase lives of 17.7 to 20.7 days (3.2 to 3.6 times the vase length of the 'White Sim' cultivar) under standard conditions (23oC, 12-h photoperiod, 70% RH). The ethylene biosynthesis pathway in these cultivars was almost completely blocked during natural senes- cence, which was responsible for the long vase life. Changes in ethylene sensitivity with flower senes- cence were evaluated using a time-lapse video recording system, which provides a simple and accurate way of evaluating ethylene sensitivity. The video system revealed that the ethylene sensitivity of vari- ous cultivars including 'Miracle Rouge' and 'Miracle Symphony' after anthesis decreased with increas- ing age.

Technical Note: Validation of a System for Monitoring Individual Feeding and Drinking Behavior and Intake in Group-Housed Cattle

The objective of this study was to validate a system for monitoring individual feeding and drinking behavior and intake in group-housed cattle. A total of 42 Holstein cows were tested with access to 24 feed bins and 4 water bins. For the purposes of this validation experiment, we focused our observations on 4 water bins and 13 feed bins. When the cow approached the feed or water bin, an antenna detected the cow's unique passive transponder and lowered the barrier, allowing the cow access to the feed or water. For each visit to the bin, the system recorded the cow number, bin number, initial and final times and weight and calculated the visit duration and intake. Bins were also monitored by direct observation and time-lapse video recording for 2 d per bin, with observations for 4 and 6 h/d for the feed and water bins, respectively. Data from direct observations were compared with the electronic data recorded by the system. Feed disappearance over 24h was assessed by using an external scale over 3 consecutive 24-h periods, and these values were compared with the sum of intakes across all visits to that bin for the same time periods. The system showed a high specificity (100%) and sensitivity (100 and 99.76% for the feed and water bins, respectively) for cow identification. The duration of the feeding and drinking visits and the feed and water intake per visit, as estimated by the monitoring system, were highly correlated with those obtained by direct observation (R2≥0.99 in all the cases). The comparison of the total feed that disappeared from each bin in 24h with the sum of the feed cows consumed from that bin during the same period differed by less than 1kg (29.92±0.90kg and 29.24±0.90kg as estimated by manual weighing and by the electronic system, respectively). This difference could be attributed to changes in feed moisture during the 24-h period. In conclusion, this electronic system is a useful tool for monitoring intakes and feeding and drinking behavior of loose-housed cows.

Cortical and cytoplasmic flows driven by actin microfilaments polarize the cortical ER-mRNA domain along the a–v axis in ascidian oocytes

Cellular mechanisms generating the polarized redistribution of maternal Type I postplasmic/PEM mRNAs in ascidian oocytes remain unknown. We have previously shown that PEM-1 mRNA is associated with a network of rough cortical Endoplasmic Reticulum (cER) polarized along the animal–vegetal (a–v) axis forming a cER–mRNA domain in mature oocytes. We now investigate the a–v polarization of this cER–mRNA domain during meiotic maturation using H. roretzi and C. intestinalis. We show that the cER and Hr-PEM-1 aggregate as interconnected cortical patches at the cell periphery before maturation, which uniformly spread out during maturation and form a reticulated organization enriched in the vegetal hemisphere at the end of maturation. Time-lapse video recordings coupled with micromanipulations reveal that stereotyped surface, cortical and cytoplasmic flows accompany the vegetal shift of the cER–mRNA domain and mitochondria-rich myoplasm. Treatments with cytochalasin B and nocodazole indicate that both polarization of the cER–mRNA domain and mitochondria-rich myoplasm and cortical and cytoplasmic flows depend on actin cytoskeleton, but not microtubules. Using cortical fragments prepared from maturing oocytes coupled with high resolution immuno/ in situ localization , we have further analyzed the effects of these inhibitors on the reorganizations the cER network and Hr-PEM-1 mRNA. We show that before maturation starts, Hr-PEM-1 mRNAs are already associated with the cER, and actin cytoskeleton inhibitors disturb their association. Finally, we hypothesize that Germinal Vesicle Break Down (GVBD) triggers an actomyosin-dependant cortical flow which directs the a–v polarization of ascidian oocytes.

Epinephrine Protects Cancer Cells from Apoptosis via Activation of cAMP-dependent Protein Kinase and BAD Phosphorylation

The stress hormone epinephrine is known to elicit multiple systemic effects that include changes in cardiovascular parameters and immune responses. However, information about its direct action on cancer cells is limited. Here we provide evidence that epinephrine reduces sensitivity of cancer cells to apoptosis through interaction with beta(2)-adrenergic receptors. The antiapoptotic mechanism of epinephrine primarily involves phosphorylation and inactivation of the proapoptotic protein BAD by cAMP-dependent protein kinase. Moreover, BAD phosphorylation was observed at epinephrine concentrations found after acute and chronic psychosocial stress. Antiapoptotic signaling by epinephrine could be one of the mechanisms by which stress promotes tumorigenesis and decreases the efficacy of anti-cancer therapies.

Rho‐kinase dependent organization of stress fibers and focal adhesions in cultured fibroblasts

The activation of Rho-kinase is known to modulate the organization of the actin-based cytoskeletal systems, including the formation of stress fibers and focal adhesions. Rho-kinase likely plays a more crucial and complex role in the organization of actin-based cytoskeletal systems than in that of myosin light chain kinase (MLCK). In order to understand the role of Rho-kinase in the organization of stress fibers and focal adhesions, we treated cultured fibroblasts with a Rho-kinase inhibitor and analyzed the stress fiber and focal adhesion organization under conventional fluorescence microscopy and replica electron microscopy. Some of the cells were transfected with GFP-labeled paxillin, actin or alpha-actinin, and the effects of the inhibitor were monitored in the living cells. The Rho-kinase inhibitor caused disassembly of the stress fibers and focal adhesions in the central portion of the cell within 1 h. However, the stress fibers and focal adhesions located in the cell periphery were not as severely affected by the Rho-kinase inhibitor. The time-lapse video recording revealed that when these cells were washed with a fresh medium in order to remove the Rho-kinase inhibitor, the stress fibers and focal adhesions located in the center of the cell gradually reorganized and, within 1.5-2 h, the cells completely recovered. This observation strongly suggests that the activation of Rho-kinase plays an important role in the organization of the central stress fibers and focal adhesions.

Dynamic motion of paxillin on actin filaments in living endothelial cells

Our three-dimensional (3-D) images showed that paxillin co-localized on actin filaments as fibrous structures, as well as clusters, in endothelial cells (ECs). In living ECs under flow condition, we monitored concurrently the intracellular dynamics of DsRed2-paxillin and GFP-actin by time-lapse video recording and dual-color fluorescence imaging. The results showed that the dynamic motion of paxillin as fibrous structures was associated with actin filaments, but not with microtubules. Our findings suggest that the actin network plays an important role not only in the assembly/disassembly of paxillin at focal adhesions, but also as a track for the intracellular transport of paxillin, which is involved in signaling pathway.

β1 Integrins Control the Formation of Cell Chains in the Adult Rostral Migratory Stream

The subventricular zone (SVZ) of the lateral ventricle is the major site of neurogenesis in the adult brain. Neuroblasts that are born in the SVZ migrate as chains along the rostral migratory stream (RMS) to the olfactory bulb. Little is known about the mechanisms that control interactions between neuroblasts during their migration. Here we show that migrating neuroblasts express beta1 integrins and that the integrin ligand laminin is localized to cell chains. Using genetically modified mice and time-lapse video recordings of SVZ explants, we demonstrate that beta1 integrins and laminin promote the formation of cell chains. Laminin also induces the aggregation of purified neuroblasts. We conclude that the formation of cell chains in the RMS is controlled in part by beta1 integrins via binding to laminin. In addition, we provide evidence that beta1 class integrins are required for the maintenance of the glial tubes and that defects in the glial tubes lead to the ectopic migration of neuroblasts into the surrounding tissue.

Circadian Rhythms in the Mating Behavior of the Cockroach, Leucophaea maderae

Mating behavior of small populations of virgin males and females of the cockroach Leucophaea maderae were continuously monitored via time-lapse video recording in controlled laboratory conditions. The time of onset of copulation was found to be rhythmic in a light cycle of 12 h light alternated with 12 h of darkness, with the peak of mating behavior occurring near the light to dark transition. This rhythm persisted in constant dim red illumination and constant temperature. In constant conditions, the period of the rhythm was slightly less than 24 h, with a peak of copulation during the late subjective day. These data demonstrated that mating behavior is gated by a circadian clock. When males and females were taken from light cycles that were 12 h out of phase, a bimodal rhythm was observed with one peak in the males' late subjective day and a second peak of equal amplitude in the late subjective day of females. The results indicated that circadian systems in both males and females contribute to the circadian rhythm in copulation. Bilateral section of the optic tracts (OTX) of both males and females abolished the rhythm, but the rhythm persisted when OTX females were paired with intact males or when OTX males were paired with intact females. Furthermore, when OTX males or OTX females were paired with intact animals that were 12 h out of phase, a bimodal rhythm was still observed. These results suggested that the circadian pacemaker in the optic lobes of both male and female cockroaches participates in the control of mating, but that a pacemaker outside the optic lobes is also likely involved. Finally, it was shown that the female's olfactory response (measured by electroantennogram) to components of the male sex pheromone exhibited a circadian rhythm, but the data suggested the peripheral olfactory rhythm is not likely to be involved in the rhythm of mating behavior.

Ultrasound and the receptivity of the endometrium

Ultrasound imaging can be used to assess the endometrium in a number of ways. In particular, time-lapse video recordings can show that the lining of the uterus undergoes rhythmical contractions that vary in strength and intensity throughout the ovarian cycle. These contractions appear to assist in sperm transport at the time of ovulation, but can decrease the chances of a fertilized egg implanting in the uterus if they persist later in the cycle. They are also the cause of ectopic pregnancies occurring in IVF treatment cycles. In order to reduce these uterine contractions and therefore increase the chances of a successful uterine pregnancy, IVF treatment cycles should incorporate minimal stimulation so as to make them as close to natural cycles as possible.

A Field Study of the Associations between Behaviors and Reproductive Performance in Lactating Sows

The present study examined factors associated with suckling, eating and postural behaviors (standing, lying and sitting) and the relationships between sow behaviors and reproductive performance. A commercial farrowing-to-finish farm with approximately 300 female pigs was visited eight times for 3 years to record sow behaviors using time-lapse video recorders for 24 hr. The lactational and postweaning reproductive performances of each sow were recorded using a recording software system. Statistical models were built to investigate the factors associated with each behavior. Regression analyses were performed to determine the associations between behavior measurements and adjusted 21-day litter weights (A21d-Wt), weaning-to-first-mating intervals (WMI) and occurrence of >or= 2 dead piglets during lactation. The means of suckling intervals and frequency of postural changes of the 145 recorded sows were 47.5 +/- 0.59 min and 86.8 +/- 2.97, respectively. As parity increased from 1 to >or= 4, the suckling interval and total standing duration increased (P<0.05). The number of nursing piglets and observation days 4 to 21 of lactation were not associated with the suckling intervals. A longer duration of lactation was associated with a longer total standing duration and a higher frequency of postural changes (P<0.05). Shorter suckling intervals were associated with heavier A21d-Wt (P<0.05). Each change in postural behavior increased the likelihood of occurrence of >or= 2 dead piglets during lactation (P<0.05). No postural or suckling behaviors were associated with WMI. Eating behavior was not associated with any reproductive performance. In conclusion, sow behaviors during lactation were related to lactational performance but were not related to postweaning reproductive performance.

Monitoring recreational activities in urban forests using long-term video observation

Summary Urban forest management needs information about the amount and kinds of recreation use, in particular about emerging recreational activities such as Nordic walking and winter bicycling. Between 2003 and 2004, year-round use levels of recreational activities were investigated in a forest in Vienna using permanent time-lapse video recording and counts by human observers on sampling days. The amount of use, yearly, weekly and daily use patterns, as well as the group size of the traditional activity groups were compared with Nordic walking and winter bicycling to characterize these activities and to better understand recreation use. For all activity groups, the rates of participation varied signifi cantly across most of the time dimensions, and the relationships between the time of day and rates of participation were different for individuals and groups. Nordic walkers displayed a temporal use pattern similar to that of jogging with workday evening and weekend morning use peaks, while winter bicycling was similar to bicycling activities with afternoon use peaks during snowy periods. Nordic walking and winter bicycling activities added to the multiuse character of the heavily used forest, but seemed not to increase the potential for user confl icts because of the current low participation rates and the Nordic walking group’s avoidance of times of heavy use.

Female mating frequency increases with temperature in two cricket species, Gryllodes sigillatus and Acheta domesticus (Orthoptera: Gryllidae)

Although the effect of temperature on calling song of male crickets has been widely studied, its influence on female mating behaviour remains largely unexplored. We examined the effect of varying temperature on female mating frequency in two cricket species ( Gryllodes sigillatus (F. Walker, 1869) (= Gryllodes supplicans (F. Walker, 1859)) and Acheta domesticus L., 1758) by providing females with multiple mating partners and recording the number of matings over 72 h intervals using time-lapse video recording. Female mating frequency increased with temperature in both species, but increased more steeply in A. domesticus than in G. sigillatus. Temperature accounted for approximately 50% of the variation in female mating frequency. These results suggest that the threshold for mating in females is temperature dependent, such that at lower temperatures only certain males are able to elicit the female mounting response required for successful mating. If temperature affects female selectivity, then male mating success in different seasons may vary, with a wider range of males gaining the opportunity to copulate at warmer times of the year. Consequently, the intensity of sexual selection may vary seasonally.

Indicators of neuritis growth and retraction in tissue culture and histological specimens

The dynamics of structural processes in cultured neurons was studied by time-lapse video recording and compared with the diversity of morphological structures in fixed preparations. Ellipsoid terminal structures indicate not the growth, but growth arrest of neurites and the start of their retraction (retraction bulbs). Only growth cones can be the indicators of neuritis growth and development. Autotomy of individual terminals is a normal process, but does not indicate neural pathology. The "neuroplasm protuberances" on fixed preparations do not indicate neuroplasm extrusion, but reflect an extreme state of retracting processes. The formation of varicosities results not from swelling, but from redistribution of the neuroplasm and often accompanies its contraction. Disruption of neural contacts and complete invagination of neurites into the cell body are not indicative of the neuron death. These cells can form new processes and make new interneuron contacts.

The use of digital video recorders (DVRs) for capturing digital video files for use in both The Observer and Ethovision

Before switching a laboratory from analog to digital, for the recording of video files for use in Noldus software such as Ethovision and The Observer, researchers need to proceed with caution. There are obvious advantages in moving to digital recording for behavioral work, including increased storage capacity; no requirement to purchase video tapes; immediate search by date, time, or event; digital images are of higher quality; ability to view study sites remotely by Internet connection; and "smart" features, such as motion detection. But before you throw away your time-lapse video recorders, time code generators, and video multiplexors, there are some important cautions to take account of. Some research groups have bought digital surveillance systems on the assumption that they work with Ethovision and The Observer, only to be disappointed. The vast majority of systems depend on proprietary compression software that must then be converted to work properly in Ethovision or The Observer.

Cortisol, behavioral responses, and injury scores of sows housed in gestation stalls

Objective: To assess welfare status of 25 pregnant sows housed in gestation stalls, in terms of cortisol concentrations, behavior responses, and injury scores. Materials and methods: Data were collected on gestation days 5, 56, and 108. Time-lapse video recording for 24-hour periods was used to observe behavior at each data point. Salivary cortisol concentrations were assessed using radioimmunoassay. Injuries were scored individually and added to provide a total injury score (TIS). Results: Cortisol concentrations were lower on gestation day 56 than on other days, and TIS was higher on day 108 than on days 5 and 56 (P &lt; .05). Time spent lying was highest on day 108 (P &lt; .05). Sows spent more time on exploration and active behavior on day 56 than on days 5 and 108 (P &lt; .05). Time for the transition from sitting to lying was higher on day 108 than on day 56 (P &lt; .05). Frequencies of overall postural change and of standing or sitting to lying and lying to sitting were highest on day 5 (P &lt; .05). Frequency of lying to standing was higher on day 5 than on day 108 (P &lt; .05). Body weight was negatively correlated with time spent on exploration and active behavior and standing, and positively correlated with time spent lying (P &lt; .05). Implications: Welfare of sows in gestation stalls appears to be more compromised during early and late stages of gestation. Providing larger sows with larger stalls might improve welfare.

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.

A primary cell culture of Drosophila postembryonic larval neuroblasts to study cell cycle and asymmetric division

Drosophila melanogaster is a key model system that has greatly contributed to the advance of developmental biology through its extensive and sophisticated genetics. Nevertheless, only a few in vitro approaches are available in Drosophila to complement genetic studies in order to better elucidate developmental mechanisms at the cellular and molecular level. Here we present a dissociated cell culture system generated from the optic lobes of Drosophila larval brain. This culture system makes it feasible to study the proliferative properties of Drosophila postembryonic Nbs by allowing BrdU pulse and chase assays, as well as detailed immunocytochemical analysis with molecular markers. These immunofluorescence experiments allowed us to conclude that localization of asymmetric cell division markers such as Inscuteable, Miranda, Prospero and Numb is cell autonomous. By time-lapse video recording we have observed interesting cellular features of postembryonic neurogenesis such us the polarized genesis of the neuroblast progeny, the extremely short ganglion mother cell (GMC) cell cycle, and the last division of a neuroblast lineage. The combination of this cell culture system and genetic tools of Drosophila will provide a powerful experimental model for the analysis of cell cycle and asymmetric cell division of neural progenitor cells.

Ethanol induces morphological and dynamic changes on in vivo and in vitro neural crest cells.

Fetal alcohol syndrome (FAS) is an embryopathology related to maternal alcohol drinking. The information concerning the factors involved in the prenatal mechanisms of ethanol action at the cellular and molecular levels is scarce. Because several abnormal changes in FAS involve regions colonized by cell lineages derived from neural crest cells (NCCs), it is reasonable to propose that epigenetic alteration of this cell population can represent an important component of the etiopathogeny. The aim of this work was to evaluate the direct effect of ethanol on a chick embryo model, as well as on in vitro NCC morphology and dynamic behavior. After ethanol treatment, in ovo or cultured chick embryos were used to determine the anatomical development of and to quantify the migratory parameters and apoptosis of NCCs. Scanning electron microscopy was performed on ethanol-perfused (and control) cultures of cephalic and trunk NCCs; the actin cytoskeleton was evaluated, and morphometric and dynamic parameters were determined after time-lapse videorecording. Recovery capacity after ethanol treatment was also determined. Chick embryos submitted to conditions sufficient to induce FAS in mammals displayed developmental disruptions frequently accompanied by cephalic/facial anomalies. In vitro studies also indicated that cephalic and trunk NCCs exposed to ethanol exhibited significant and permanent changes regarding cell shape, surface morphology, apoptotic cell death, cytoskeleton, and distance and velocity traveled, as well as an abnormal pattern of migration. Taking into account that even a limited period of abnormal behavior may imply serious consequences in the final cues of an embryonic cell population, our results indicate that the biological effects of ethanol on early development-even during a short time-could induce permanent ontogenetic perturbations of NCCs, with potentially dramatic effects on embryonic morphogenesis. These results support an important participation of NCCs in the etiopathogeny of FAS.

Transparent Polymeric Cell Culture Chip with Integrated Temperature Control and Uniform Media Perfusion

Modern microfabrication and microfluidic technologies offer new opportunities in the design and fabrication of miniaturized cell culture systems for online monitoring of living cells. We used laser micromachining and thermal bonding to fabricate an optically transparent, low-cost polymeric chip for long-term online cell culture observation under controlled conditions. The chip incorporated a microfluidic flow equalization system, assuring uniform perfusion of the cell culture media throughout the cell culture chamber. The integrated indium-tin-oxide heater and miniature temperature probe linked to an electronic feedback system created steady and spatially uniform thermal conditions with minimal interference to the optical transparency of the chip. The fluidic and thermal performance of the chip was verified by finite element modeling and by operation tests under fluctuating ambient temperature conditions. HeLa cells were cultured for up to 2 weeks within the cell culture chip and monitored using a time-lapse video recording microscopy setup. Cell attachment and spreading was observed during the first 10-20 h (lag phase). After approximately 20 h, cell growth gained exponential character with an estimated doubling time of about 32 h, which is identical to the observed doubling time of cells grown in standard cell culture flasks in a CO2 incubator.