Virtual nitrogen tank to monitor frozen cell stocks

We developed a program to facilitate the monitoring of biological samples (cell lines, sera, etc.) that are stored in liquid nitrogen containers. The program consists of a "virtual" container in which scientists can store their samples and a program that records the location of each sample, cell characteristics, storage dates, names of the manipulators and much more. Additional comments and a photograph can be associated with each vial, allowing for reliable tracking of samples. Vials can then be identified according to any parameter of interest to the scientist, including associated comments. Once identified, the program visually presents the location of these vials, which simplifies retrieving them from the real container. The program records the thawing of vials, along with the date and the name of the operator. Any academic laboratory requesting this standalone program will be granted a free license for its use.     

Habitat stability and the larval mosquito community in treeholes and other containers on a temperate Island

The temporal fluctuation of water levels and the presence of mosquito larvae were investigated for four types of small container habitats (treeholes, bamboo stumps, cemetery stone vases, and cemetery stone vessels) on Kabeshima Island in Kyushu, southwestern Japan. The probability that containers held water was positively correlated with the quantity of the preceding rainfall and with the depth and volume of the containers. It was estimated that dehydration occurred more regularly in autumn and winter than in summer. The probability that mosquito larvae were present in each type of container was positively correlated with habitat stability in terms of the probability of the existence of standing water and the coefficient of variation of the water level. Twelve species of mosquito larvae, including two rare predators, were found. Species composition differed between the different types of container. Although 4 to 10 species used each type of container, the median number of species per container was two for treeholes and bamboo stumps, and one for the others. The dominant species wasTripteroides bambusa in treeholes and bamboo stumps,Aedes albopictus in stone vases, andA. japonicus in stone vessels. The larval mosquito community, which lacked major predators, possessed the following features that may facilitate the coexistence of many species: (1) niche segregation amongst species in terms of their selection of container types; (2) an aggregated distribution of the individual species among containers of the same type; (3) high intraspecific mean crowding (and hence probably intense intraspecific competition) in the dominant species in each type of container; (4) independent species associations within the same type of container; and (5) low interspecific mean crowding (and hence probablynot intense interspecific competition) between species in the same type of container.     

Population dynamics of the immature stages of Aedes aegypti (Diptera: Culicidae), vector of dengue: a longitudinal study (1996-2000)

A four year study was conducted on a natural population of immature stages of Aedes aegypti after the re-invasion of Argentina by this vector in 1987. Thirty six plastic containers with 700 ml of dechlorinated water were placed in the La Plata Zoological Garden, La Plata, Argentina. A strip of filter paper around each container was added to facilitate egg counting. Eggs, larvae and pupae were counted weekly in each container from September, 1996 to August, 2000. After egg counting, papers were submerged to facilitate egg hatching and a new paper was placed in each container. Presence of A. aegypti immature stages was recorded from December-January to June during each of the four years of this study. In 1997, 13,105 eggs, 7,978 larvae and 1,476 pupae were registered with 54.7 % positive containers; during 1998, 8,194 eggs, 668 larvae and 142 pupae were recorded with 28.3 % positive containers; 13,510 eggs, 3,690 larvae and 743 pupae were registered during 1999 with 56.7 % positive containers; and 16,327 eggs, 4,669 larvae and 715 pupae during 2000 with 59.3 % of containers with presence of A. aegypti. Egg number and hatching rate were drastically reduced in 1998 when temperatures from December to May were 1 to 2.5 degrees C lower than the other years of this study. These colder than usual temperatures in the summer of 1998 were a consequence of the El Ni?o event.     

Comparison of three containers used for the transport of cooled stallion semen

Three containers commonly used to transport cooled equine semen (Equitainer, ExpectaFoal and a Swedish-designed semen-transport container, previously called the Salsbro Box and now called Equine Express) were compared, using four ejaculates from each of three stallions. Each ejaculate was diluted to a spermatozoal concentration of 25 x 10(6)/ml with a nonfat dry milk-glucose extender containing amikacin sulfate (1 mg/ml) and potassium penicillin G (1000 units/ml). Extended semen was divided into three 40-ml aliquots for placement in each of the three semen-transport containers. The extended semen was stored in the containers for 24 h prior to analysis. Stored semen was warmed for 15 min at 37 degrees C, then video records of sperm motility were obtained for evaluation using a Hamilton-Thorne motility analyzer equipped with a stage warmer set at 37 degrees C. The temperature of 40-ml aliquots of semen extender stored in each container was also measured for 60 h using a copper-constantan thermocouple placed in the center of the stored samples. Intervals from onset of storage until sample temperature exceeded 10 degrees C during the warming phase were 27.5, 33.5 and 53 h, for the Expecta-Foal, Equine Express and Equitainer, respectively. Semen extender stored in the Equitainer compared most favorably to ideal cooling rates and storage temperatures published previously. Following a 24-h storage period, the mean percentages of motile, progressively motile, and rapidly motile spermatozoa, as well as the mean spermatozoal curvilinear velocity were similar (P > 0.05) among the three containers.     

The effect of container porosity on root environment and plant growth

Summary The effect of container porosity on water relations in the substrate and on plant growth was examined. With clay containers the proportional effect of the wall on water loss varied with season, being responsible for 42 per cent of the total loss in winter and 25 per cent in summer. Water loss from plastic containers was 55 per cent of that from clay containers in winter, and 66 per cent in summer. Wall porosity also had a significant effect on water distribution within the container under high radiation conditions. Tomato plants grown under three water regimes showed no effect of matric stress in winter, but growth was significantly reduced for each increase in stress in summer. There was no interaction between container type and water stress. Non-porous containers gave a higher water-use efficiency and had a lower irrigation requirement than porous ones, their greatest advantage being under conditions of low transpirational loss and high evaporative loss.     

Added-value of mosquito vector breeding sites from street view images in the risk mapping of dengue incidence in Thailand

Dengue is an emerging vector-borne viral disease across the world. The primary dengue mosquito vectors breed in containers with sufficient water and nutrition. Outdoor containers can be detected from geotagged images using state-of-the-art deep learning methods. In this study, we utilize such container information from street view images in developing a risk mapping model and determine the added value of including container information in predicting dengue risk. We developed seasonal-spatial models in which the target variable dengue incidence was explained using weather and container variable predictors. Linear mixed models with fixed and random effects are employed in our models to account for different characteristics of containers and weather variables. Using data from three provinces of Thailand between 2015 and 2018, the models are developed at the sub-district level resolution to facilitate the development of effective targeted intervention strategies. The performance of the models is evaluated with two baseline models: a classic linear model and a linear mixed model without container information. The performance evaluated with the correlation coefficients, R-squared, and AIC shows the proposed model with the container information outperforms both baseline models in all three provinces. Through sensitivity analysis, we investigate the containers that have a high impact on dengue risk. Our findings indicate that outdoor containers identified from street view images can be a useful data source in building effective dengue risk models and that the resulting models have potential in helping to target container elimination interventions.     

Type of closure prevents microbial contamination of cosmetics during consumer use.

The dispensing closure used for containers plays an important role in protecting cosmetics from in-use microbial contamination. This hypothesis was tested by aseptically packing unpreserved shampoo and skin lotion into containers with three different closure types which provided various degrees of protection against consumer and environmental microbial insults. Shampoo was packed in containers with slit-cap (n = 25), flip-cap (n = 25), or screw-cap (n = 28) closures. Skin lotion was packed in containers with pump-top (n = 21), flip-cap (n = 18), or screw-cap (n = 21) closures. The products were then used by volunteers under actual in-use conditions for 3 (shampoo) or 2 (skin lotion) weeks. After use, the products were evaluated for microbial contamination by using standard methods for enumeration and identification. The standard screw-cap closure provided only minimal protection against microbial contamination of both the shampoo (29% contamination incidence) and the skin lotion (71%). The slit-cap closure on the shampoo container and the flip-cap closure on the skin lotion container provided slightly enhanced degrees of protection (21 and 39% contamination incidence, respectively). The greatest amount of protection (i.e., lowest contamination incidence) was provided by the flip-cap closure for the shampoo container (0%) and the pump-top closure for the skin lotion container (10%). As a result, closure type plays an important role in protecting poorly preserved products from in-use microbial contamination.     

Type of closure prevents microbial contamination of cosmetics during consumer use

The dispensing closure used for containers plays an important role in protecting cosmetics from in-use microbial contamination. This hypothesis was tested by aseptically packing unpreserved shampoo and skin lotion into containers with three different closure types which provided various degrees of protection against consumer and environmental microbial insults. Shampoo was packed in containers with slit-cap (n = 25), flip-cap (n = 25), or screw-cap (n = 28) closures. Skin lotion was packed in containers with pump-top (n = 21), flip-cap (n = 18), or screw-cap (n = 21) closures. The products were then used by volunteers under actual in-use conditions for 3 (shampoo) or 2 (skin lotion) weeks. After use, the products were evaluated for microbial contamination by using standard methods for enumeration and identification. The standard screw-cap closure provided only minimal protection against microbial contamination of both the shampoo (29% contamination incidence) and the skin lotion (71%). The slit-cap closure on the shampoo container and the flip-cap closure on the skin lotion container provided slightly enhanced degrees of protection (21 and 39% contamination incidence, respectively). The greatest amount of protection (i.e., lowest contamination incidence) was provided by the flip-cap closure for the shampoo container (0%) and the pump-top closure for the skin lotion container (10%). As a result, closure type plays an important role in protecting poorly preserved products from in-use microbial contamination.     

EFFECTS OF DAY AND NIGHT TEMPERATURE AND TYPE OF CONTAINER ON THE GROWTH OF F1 HYBRID ANNUALS IN CONTROLLED ENVIRONMENTS

The effects of three day/night temperatures (18/12 C, 24/18 C, and 30/24 C), and two types of containers (clay and plastic) on the seedling growth of three F₁ hybrid annuals were determined after 14 days of controlled-environment treatment. A day/night temperature of 18/12 C was severely limiting to the early seedling growth of ‘Blue Blazer’ ageratum (Ageratum houstonianum Mill.), ‘Pink Cascade’ petunia (Petunia hybrida Vilm.), and ‘Double Eagle’ marigold (Tagetes erecta L.). Raising the day/night temperature to 24/18 C resulted in striking increases in growth in both clay and plastic pots. Fresh and dry weights of tops for all three species were increased four- to fivefold, and leaf areas of ageratum and petunia were increased more than fivefold irrespective of container. Height and node number were also increased significantly but to a much lesser degree. A day/night temperature of 30/24 C appeared to be optimum for early seedling growth of F₁ hybrid annuals the optimum being based on dry matter accumulation, stem elongation, node development, and leaf area production. Growth of lateral shoots was greatly stimulated at 30/24 C, especially in marigold. There were no appreciable differences in fresh or dry weight of tops or in leaf area between plants in clay and plastic containers at any of the three day/night temperatures.     

Successful vitrification of bovine blastocysts on paper container

Cryopreservation of bovine embryos can be performed by a variety of methods with variable degree of success. Here, we report a new, easy to perform, simple, inexpensive, and successful method for vitrification of bovine blastocysts. In vitro produced bovine blastocysts were exposed to vitrification solution (5.5 m ethylene glycol, 10% serum and 1% sucrose) in one single step for 20 s, loaded on a paper container prepared from commonly available non-slippery, absorbent writing paper, and then were directly plunged into liquid nitrogen for storage. Vitrified blastocysts were warmed by serial rinsing in 0.5, 0.25 and 0.125 m sucrose solution for 1 min each. Results showed that one step exposure of bovine blastocysts to cryoprotective agents was sufficient to achieve successful cryopreservation. Under these conditions, more than 95% of blastocysts survived the vitrification-warming on paper containers which was significantly higher than those obtained from other containers, such as electron microscope (EM) grid (78.1%), open pulled straw (OPS; 80.2%), cryoloop (76.2%) or plastic straw (73.9%). Embryo transfer of blastocysts vitrified-warmed on paper container resulted in successful conception (19.3%) and full-term live birth of offspring (12.3%) which were lower (P < 0.05) than those obtained from non-vitrified blastocysts (38.0 and 32.7%) but were comparable (P > 0.05) to those obtained from blastocysts vitrified-warmed on EM grid (23.3 and 14.2%). Our results, therefore, suggest that paper may be an inexpensive and useful container for the cryopreservation of animal embryos.     

Aeration time following ethylene oxide sterilization for reusable rigid sterilization containers: concentration of gaseous ethylene oxide in containers

Because ethylene oxide (EO) gas is toxic to humans, restrictions have been imposed on its use for sterilization, specifying allowable levels of residual EO remaining in sterilized apparatus and materials. However, the aeration time that optimizes the removal of the remaining EO when a rigid sterilizing container is used for a vessel had not been identified. Therefore, polyvinyl chloride, which easily adsorbs EO, was placed in rigid sterilizing containers, and aeration was carried out after 1, 8, 12, 17, and 24 hours. After standard EO sterilization, the EO concentrations remaining in the air in the rigid containers were measured. The results indicate that a period of 17 hours of aeration is appropriate when a rigid sterilizing container is used.     

Effect of temperature on longevity and infection with the nematode Strelkovimermis spiculatus (Nemata: Mermithidae), a parasite of mosquitoes

Strelkovimermis spiculatus is a common parasite of culicid species in Argentina. Effect of temperature on longevity and infectivity of juvenile preparasites of S. spiculatus was determined at 4, 10, 20 and 27 degrees C. Three containers with 100 ml of dechlorinate water and 300 preparasites (12 hour-old), were placed for each day and temperature, during 40 days (total = 480 containers). Survived preparasites were counted on 12 containers per day (three for each temperature). When number of survived preparasites was determined, second instar larvae of Aedes aegypti were added to each container in a 10:1 ratio (preparasites:mosquito) to determine infectivity of daily survived preparasites. Longevity of preparasites decreased at higher temperatures. Maximum longevity of preparasites maintained at 4, 10, 20 and 27 degrees C were 35, 30, 25 and 27 days, respectively. Survivorship of preparasites, exposed to the same temperatures, varied from 57% to 100% at day two, from 21% to 77% at day five and from 9% to 33% at day ten. Infectivity of preparasites maintained at temperatures from 4 to 27 degrees C was always higher than 70%. Extended longevity with maintenance of the infectivity capacity of preparasites, are important attributes to consider S. spiculatus an effective mean of controlling a large number of culicid species between 4 and 27 degrees C.     

Improved method of studying the blood for sterility

The technique used for the inoculation and subculturing of blood samples in testing them for sterility is described. This technique eliminates the possibility of contaminating the culture medium and the blood sample under test with extraneous bacterial flora. Blood samples were inoculated without opening the containers with the culture medium. Inoculation was made with the syringe and the needle used for taking the blood sample through the punctured rubber stopper closing the container. Subculturing on solid culture media was also carried out without opening the containers: the rubber stopper was punctured and the contents of the container withdrawn with a pipette needle. The use of this new technique made it possible to detect bacteremia in 12.8% of cases, only in persons with purulent and septic diseases, whereas by using the existing technique bacteremia was detected both in sick and healthy persons, in 38.6% and 26.6% of cases, respectively.     

Stability of containerized urban street trees

Trees are usually grown in containers in the nursery until they reach a certain size, whereupon they are transplanted to a permanent location. Infrastructure development has often led to the removal of large trees. To maintain lush foliage and trees of a size that benefit urban ecology, trees can be grown in containers. Containerized trees can be moved from one location to another, and this relocation does not require root pruning or crown-size reduction. The drawback to having trees in containers is the small and confined volume of the container, which limits tree root development and thus affects containerized tree stability. The objective of this study was to understand the failure mechanisms for and the effect of the root dimensions on the stability of containerized trees. Therefore, small-scale stability model tests were conducted which were verified using numerical and analytical models. The results identified two failure modes that were likely to occur: tree overturning and container overturning. The mode of failure was dependent on the root dimensions. When the trees had extended their roots deep into the container, the whole container would overturn in the event of failure due to increased root confinement and shear resistance of the soil. On the other hand, the main failure mechanism when there was shallow root development was the uplifting of the tree from the container while the container remained upright. The results from numerical and analytical models were consistent with those obtained during the small-scale model stability tests.     

长白山阔叶红松林凋落物组成及其季节动态

为了解群落尺度上凋落物组成及其时空变化,对长白山阔叶红松林25 hm² 样地内2008年度收集的凋落物进行统计分析.结果表明: 一年间,150个收集器内收集的凋落叶分属35种树木,占样地内胸径≥1 cm树种数(52种)的67.3%;凋落物总量为29.39 kg,折合3918.4 kg·hm^-2,其中,阔叶、杂物、针叶和枝条凋落量分别占凋落物总量的61.7%、18.0%、11.7%和 8.6%;紫椴、水曲柳、蒙古栎、色木槭和春榆5个树种的叶凋落量占阔叶总凋落量的83.8%;不同树种的凋落量季节差异很大,61.9%的凋落物产生于9月13日至10月10日.其中,红松和紫椴叶凋落高峰出现在9月13-26日,蒙古栎、春榆和色木槭叶凋落高峰出现在9月27日-10月10日.收集器间凋落物量差异较大,其中68个收集器的年凋落量在150～200 g,1个收集器大于500 g;单个收集器全年最多可收集到18个树种的凋落叶,凋落叶种数为12种的收集器最多(32个).叶凋落量与样地内该树种的胸高断面积总和成正比.样地内凋落叶的分布存在明显的空间异质性,且收集器内凋落物的收集量与其所处的位置有关.     

Do container size and predator presence affect Culex (Diptera: Culicidae) oviposition preferences?

Organisms with complex life cycles typically do not exhibit parental care. Hence, the ability of adult females to choose quality oviposition sites is critical for offspring success. Gravid females of many insect taxa have the capability to detect environmental conditions in water-holding containers (e.g., resource level, presence of competitors or predators) and to choose the sites that are most suitable for offspring growth and development. Mosquitoes may also detect physical container characteristics related to water permanence such as surface area, volume, or container size, and some species such as those in the genus Culex have been shown to prefer larger containers. However, predators may also preferentially colonize larger containers; thus, ovipositing females may face decisions based on cues of site quality that balance the costs and benefits for offspring. We used a field experiment to evaluate the oviposition preferences of two Culex species in response to experimental container size and predator abundances within the containers. We found that both species avoided ovipositing in the largest containers, which have high abundances of Chaoborus sp. and dragonfly larvae (predators). However, the container size most commonly chosen for oviposition (15-L buckets) also had high mean abundance per liter of dragonfly larvae. These results suggest either prey naiveté or reduced vulnerability of these species to dragonflies compared to Chaoborus sp. Other potential mechanisms for the observed patterns are discussed.     

Linking Mosquito Infestation to Resident Socioeconomic Status, Knowledge, and Source Reduction Practices in Suburban Washington, DC

Eliminating water-holding containers where mosquitoes oviposit and develop (source reduction) can help manage urban disease-vector mosquitoes. Source reduction requires residents to be knowledgeable of effective practices and motivated to implement them. We tested relationships between demographics, resident knowledge, attitudes, and practices (KAP), and mosquito infestation by administering larval mosquito surveys and KAP questionnaires in Washington, DC. Respondents who reported practicing source reduction had lower numbers of pupae-positive containers and Culex pipiens-positive containers, but not Aedes albopictus-positive containers or water-holding containers, in their yards. When controlling for numbers of water-holding containers in statistical models, residents who reported source reduction had lower numbers of A. albopictus-positive containers in addition to numbers of pupae-positive containers and C. pipiens-positive containers. These results suggest that while active container reduction may be effective at reducing C. pipiens and overall pupal production, it may be offset by other resident activities that add containers to yards, and that source reduction that involves mosquito habitat management without outright container removal can also be effective at reducing A. albopictus. Source reduction was related to respondent knowledge of mosquitoes and, in particular, specific knowledge of mosquito development, which both varied with demographics alongside respondent motivation to control mosquitoes. Respondents from high socioeconomic status households reported greater knowledge but lower motivation than respondents from middle and low socioeconomic-status households. We conclude that mosquito-related education will help promote community-based container management as part of integrated mosquito management programs, particularly in middle and low socioeconomic status neighborhoods with lower knowledge and high motivation.     

Differences in preservation of canine chilled semen using different transport containers

In the present study, the effect of three different containers in the preservation of dog chilled semen, during 24, 48 and 72h was evaluated. Weekly sperm pools of different dogs were obtained, during 10 consecutive weeks. Semen samples were diluted in egg-yolk-Tris-fructose extender and stored in a Styrofoam box, a common Thermos flask and an Equitainer. Progressive motility, morphology and sperm membrane integrity were examined in semen aliquots taken daily from each container during the 3 days of storage. Additionally, integrity of the acrosome and sperm plasma membranes, determined by PI/Fitc-PSA staining was assessed at 48 and 72h of storage. At 24h no differences were observed between the three containers for the evaluated parameters. At 48h samples kept in the Equitainer presented a higher progressive motility than samples kept in the Thermos. At 72h, progressive motility was higher in the Equitainer than in the other two containers. Only samples kept in the Equitainer maintained similar levels of progressive motility between 24 and 72h. Membrane integrity assessed by eosin-nigrosin deteriorated over the 72h period, whereas functional membrane integrity determined by the hypoosmotic swelling test was independently affected by type of container (the Equitainer) kept a higher percentage of sperm cells with intact membrane) and time of storage (a decrease of membrane integrity between 24 to 72h). Staining with PI-Fitc-PSA allowed the detection of differences between containers but not between the two studied storage periods (48 and 72h). The results indicated that the use of the Equitainer is preferable when transporting chilled dog semen for more than 48h.     

Effects of transport container and ambient storage temperature on motion characteristics of equine spermatozoa

This study was conducted to compare the cooling rates and storage temperatures within equine semen transport containers exposed to different ambient temperatures, and to evaluate the ability of these containers to preserve spermatozoal motility following 24 h of storage under these conditions. In Experiment 1, nonfat dried milk solids, glucose, sucrose, equine semen extender was divided into seven 40-mL aliquots and loaded into seven different semen transport containers: Equitainer I, Equitainer II, Equitainer III, ExpectaFoal, Bio-Flite, Lane STS, and Equine Express. After containers were loaded, they were subjected to one of three ambient storage temperatures: 1) 22 degrees C for 72 h, 2) -20 degrees C for 6 h followed by 22 degrees C for 66 h, or 3) 37 degrees C for 72 h. Cooling rates and storage temperatures of semen extender in each container were monitored with thermocouples and a chart recorder. In Experiment 2, semen from each of three stallions (3 ejaculates per stallion) was diluted to 25 x 10(6) spermatozoa/mL with semen extender, divided into 40 mL aliquots and loaded into transport containers as in Experiment I. Containers were subjected to one of three ambient storage conditions: 1) 22 degrees C for 24 h, 2) -20 degrees C for 6 h, followed by 22 degrees C for 18 h, or 3) 37 degrees C for 24 h. After 24 h of storage, spermatozoal motion characteristics (percentage of motile spermatozoa; MOT, percentage of progressively motile spermatozoa; PMOT, and mean curvilinear velocity; VCL) were evaluated using a computerized spermatozoal motion analyzer. Significant interactions were detected among storage conditions and semen transport containers for the majority of the temperature endpoints measured. When exposed to temporary ambient freezing conditions, the lowest temperatures attained by samples in containers ranged from -2.8 to 0.8 degrees C. Lowest temperature samples attained was not correlated (P > 0.05) with spermatozoal motility under any ambient condition. However, time below 4 degrees C was highly correlated (P < 0.05) with a reduction in spermatozoal motility. Mean cooling rates from 20 degrees C to 8 degrees C did not correlate with spermatozoal motility, except when containers were exposed to temporary freezing conditions. No container cooled samples below 6 degrees C in 22 degrees C or 37 degrees C environments except for the ExpectaFoal, in which samples fell below 4 degrees C under all ambient conditions. Ambient temperature affected MOT, PMOT and VCL of semen stored in all containers (P < 0.05) except for the Equitainer II in which motion characteristics remained high and were similar among all ambient temperatures (P > 0.05). Results suggest that stallion semen may be able to tolerate a wider range of cooling rates and storage temperatures than previously considered safe.     

Pricing storage of outbound containers in container terminals

In container yard of container terminals, a storage charge is imposed to encourage customers to limit the space required for their containers. This study addresses the storage price scheduling problem for the storage of outbound containers. The price schedule consists of the free-time limit, which is the maximum duration for a container to stay in the yard without any charge, and storage charge per day for storing a container past the free-time-limit. Empirical data suggests that the efficiency of loading operations significantly depends on the space utilized by a vessel’s outbound containers. Mathematical models are developed to determine the optimal storage price schedule in such a manner that the terminal’s total profit is maximized or the total system’s cost is minimized. Both single and multi-vessel cases are considered. Properties of the optimal solution are derived from the mathematical models and numerical experiments are conducted to validate solutions.