Sterilization by Means of Hydrochloric Acid Vapour

As heat sterilization of glass bottles often results in breakage and of most plastic containers in deformation, we investigated a rapid low-temperature sterilization method as an alternative. Vapour evolving from hydrochloric acid was chosen because it does not leave toxic residues which might contaminate food packed in treated containers. Vapour evolving from 0.25 ml of 31% (w/w) hydrochloric acid reduced the number of viable spores of aerobic and anaerobic bacteria in bottles (300 ml) by a factor of at least 2500 and that from 0.01 ml of 37% (w/w) hydrochloric acid reduced the number of mould spores by a factor of over 40 000 within 5 min at 20°C.     

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.     

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.     

Effects of media and the presence of bovine oviduct epithelial cells during in vitro fertilization on fertilizability and developmental capacity of bovine oocytes

The effect of the presence of bovine oviduct epithelial cells (BOEC; Experiment 1) as well as the effects of media (Tyrode fertilization medium: TFM vs synthetic oviduct fluid: SOF), fertilization containers (drops in petri dish vs 96-wells), and the number of oocytes per drop and well (5 vs 10) for in vitro fertilization (Experiment 2) on the fertilizability and in vitro development of bovine oocytes were investigated. Immature oocytes with cumulus cells were cultured in TCM199 supplemented with 10% ECS and 2.5x10(6) granulosa cells for 24 hours at 39 degrees C under 5% CO(2) in air. In vitro fertilization was performed with frozen-thawed, heparin-treated spermatozoa (100 mug/ml, 15 minutes) and with BOEC (Experiment 1). In Experiment 2, in vitro fertilization was performed with two different media (TFM and SOF) and various conditions (culture dish and different number of oocytes). Cleavage, development to the blastocyst stage were evaluated on Day 2 and Day 7 after the start of culture. Effect of the presence of BOEC on fertilizability and developmental capacity (Experiment 1) was not significantly different. In Experiment 2, alterations in media, containers and number of oocytes during in vitro fertilization had no affect. The SOF medium showed results similar to those of TFM (normal fertilization rate: 63.2 vs 64%; cleavage: 69.3 vs 73.9%; development to the blastocyst stage: 14 vs 15%; and mean number of nuclei per blastocyst: 80.5 vs 86.6). The results indicate that the presence of BOEC during in vitro fertilization did not improve fertilizability, and that SOF as well as TFM medium can be utilized as a simple fertilization medium.     

细胞工厂在轮状病毒基因重配株LD9培养中的应用初探

为了探索用细胞工厂代替转瓶培养轮状病毒基因重配株LD9及收获高滴度的LD9病毒原液和提高产量的可行性,分别在2层4、层细胞工厂和3L1、5L转瓶培养Vero细胞,比较两种容器内细胞的生长状态。结果显示,以相同活细胞数2.5×104/ml同时接种两种不同培养容器时,细胞工厂培养3d已长成单层,而转瓶培养需5d;对两种容器长满单层时的细胞经胰酶消化后通过细胞仪计数、分析,结果显示,两种容器培养细胞长成单层时的单位面积细胞密度相当;对长成致密单层细胞的两种容器以相同的MOI(MOI=0.1)接种LD9病毒,转瓶培养的病毒于第8d病毒滴度达到高峰,为6.0～6.5 lgCCID50/ml;细胞工厂第5d病毒滴度达高峰,为6.5 lgCCID50/ml,并于第9d病毒滴度再次达到峰值,为6.0～6.5 lgCCID50/ml,实现二次收获病毒。     

Child-resistant containers: are we kidding ourselves?

A 20-month-old child was accidentally poisoned after biting through the bottom of a medicine container and ingesting the tablets inside. Consequently a study was carried out to determine the force required to fracture 20 randomly selected 25 and 32 ml polystyrene containers to see whether this exceeded the bite force of a child''s jaw. Tests were performed at displacement rates of 0.5 and 10 cm/min. All the containers failed at well below the bite force recorded for children, which is 392 N. All containers must conform to a British Standards test that requires that they withstand a force of only 35 N. Clearly this is not enough to safeguard small children. The use of polystyrene containers should be scrutinised closely, as the case of accidental poisoning reported may not be unique.     

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.     

Laboratory feeding and egg production in the chaetognath Sagitta crassa Tokioka

Egg production, daily ration, and growth efficiency in Sagitta crassa Tokioka are described along with the morphological characteristics of the ovary and body during laboratory observations. Feeding by S. crassa in small containers containing 50 or 100 ml sea water and feeding rates bore a linear relationship to food concentration from 5 to 20 copepods per 100 ml. Among 100 animals individually isolated in containers, 3 animals were the longest lived and the best egg producers. One of these produced eggs on more than 30 consecutive days, producing almost 1000 eggs. Egg production showed periodic change at intervals from 7 to 10 days. Seminal vesicles also became periodically filled or empty. Fluctuations in ovary length and morphological changes of ovary and body were observed through life. Daily ration of S. crassa ranged from 8.7 to 10.4 prey per day or ≈ 60 μg in dry wt. The specific daily ration ranged from 0.347 to 0.568. S. crassa had a growth efficiency (dry-wt basis) which increased during the early life of the animal (≈28%), then decreased with age (22 → nearly 0%). Reproductive efficiency ranged from 7.0 to 16.4%, with a mean of 12.5%.     

Suitability of containers from different sources as breeding sites of Aedes aegypti (L.) in a cemetery of Buenos Aires City,Argentina

Cemeteries are ideal urban areas to study the importance of different types of containers as breeding sites of Aedes aegypti (L.). In the present study, the suitability of plastic, glass, ceramic and metal containers was evaluated in four patches within a cemetery of Buenos Aires City, Argentina. Between October 1998 and May 2000, we found 215 breeding sites of Ae. aegypti out of 13,022 water-filled containers examined. In two patches containing microenvironments sheltered from the sun, the use of the different types of containers was proportional to the offer (correlation coefficient = 0.99, P < 0.05 in both cases). In the remaining patches, plastic and metal containers were the most and less frequent breeding sites, respectively (P < 0.001 in both cases). The number of immatures per breeding site (median = 4.5) did not show significant differences among the four types of containers examined (H3, 215 = 1.216, P = 0.749). Differences found in patches from a same cemetery suggest that different microenvironmental conditions affect the suitability of each type of container for Ae. aegypti breeding. Plastic containers appeared as key breeding sites that should be removed to reduce the Ae. aegypti population in the study area.     

Environmental evaluation of aluminium cans for beverages in the German context

Background  

In the years 2000 and 2002, the German Environment Agency in Berlin (UBA) published the results of a comprehensive LCA study on beverage containers comprising aluminium cans with volumes of 330 ml and 500 ml. Starting with the aluminium can scenarios and the respective results obtained during the UBA study, additional analyses were performed by IFEU in 2003, a German consultant having been a member of the project team working on the UBA study. The objective was to examine the influence of selected parameters on the LCA profile of carbonated soft drink containers. Data and method were in complete analogy with the LCI and LCA part of the UBA study.     

EXPRESSION OF THE MITOCHONDRIAL GENOME IN HELA CELLS : XV. Effect of Inhibition of Mitochondrial Protein Synthesis on Mitochondrial Formation

The effect of selective inhibition of mitochondrial protein synthesis by chloramphenicol at 40 or 200 µg/ml on the formation of mitochondria in HeLa cells was investigated. HeLa cells, under the conditions used in the present work, grow at a decreasing rate for at least four cell generations in the presence of 40 µg/ml chloramphenicol, and for two generations in the presence of 200 µg/ml chloramphenicol. The progressive cell growth inhibition which begins after 2 days of exposure of the cells to 40 µg/ml chloramphenicol is immediately or gradually reversible, upon removal of the drug, for periods up to at least 8 days of treatment, though there is a progressive loss of cloning efficiency. In cells which have been treated for 6–7 days with 40 or 200 µg/ml of chloramphenicol, mitochondrial protein synthesis occurs at a normal or near-normal rate 1 h after removal of the drug. Mitochondria increase normally in number and show a normal size and amount of cristae in the presence of either concentration of drug. However, in 4–5% of the mitochondrial profiles the cristae appear to be arranged in unusual, circular, looped or whorled configuration.     

Dynamics and Characterization of Aedes aegypti (L.) (Diptera: Culicidae) Key Breeding Sites

The present study aimed to analyze the dynamics of containers used as breeding sites by Aedes aegypti (L.) in the city of Aracaju, SE, one of the Northeast Brazilian states. A total of three entomological surveys were performed during different precipitation levels. Breeding sites were categorized according to their function into storage, disposable containers, and reusable containers. “Mean number of pupae” and “frequency of each type of breeding site” were the criteria considered to identify key breeding sites. House index and Breteau index were calculated in each survey. A total of 3,647 water reservoirs were found, of which 220 were breeding sites, where 22,880 immature forms were identified. There were no differences in the mean number of larvae of several types of breeding sites and in the number of larvae among surveys. Larval indices showed a reduction in the second visit, but with no effect on adult occurrence when the number of pupae was considered. Key breeding sites resulted from containers used for water storage. The area studied showed conditions favorable to a new epidemic of dengue fever.     

ROLE OF THE MITOCHONDRIAL GENOME DURING EARLY DEVELOPMENT IN MICE : Effects of Ethidium Bromide and Chloramphenicol

The role of the mitochondrial genome in early development and differentiation was studied in mouse embryos cultured in vitro from the two to four cell stage to the blastocyst (about 100 cells). During this period the mitochondria undergo morphological differentiation: progressive enlargement followed by an increase in matrix density, in number of cristae, and in number of mitochondrial ribosomes. Mitochondrial ribosomal and transfer RNA synthesis occurs from the 8 to 16 cell stage on and contributes to the establishment of a mitochondrial protein-synthesizing system. Inhibition of mitochondrial RNA- and protein-synthesis by 0.1 µg/ml of ethidium bromide or 31.2 µg/ml of chloramphenicol permits essentially normal embryo development and cellular differentiation. Mitochondrial morphogenesis is also nearly normal except for the appearance of dilated and vesicular cristae in blastocyst mitochondria. Such blastocysts are capable of normal postimplantation development when transplanted into the uteri of foster mothers. Higher concentrations of these inhibitors have general toxic effects and arrest embryo development. It is concluded that mitochondrial differentiation in the early mouse embryo occurs through the progressive transformation of the preexisting mitochondria and is largely controlled by the nucleocytoplasmic system. Mitochondrial protein synthesis is required for the normal structural organization of the cristae in blastocyst mitochondria. Embryo development and cellular differentiation up to the blastocyst stage are not dependent on mitochondrial genetic activity.     

MUCILAGE-PRODUCING CELLS IN THE SEED COAT OF PLANTAGO OVATA: DEVELOPMENTAL FINE STRUCTURE

As the ovule of Plantago ovata matures into a seed its epidermal cells are transformed from undifferentiated parenchyma to thin-walled containers of almost pure mucilage. During this process the volume of the cells increases 60–80 fold, and the protoplast degenerates to a remnant. Rapid cell expansion begins with pollination and is accompanied by an increase in the size of the nucleus and nucleolus, a change in the random arrangement of ribosomes, a decrease in the thickness of cell walls, and synthesis of starch. Deposition of mucilage inside vacuoles and between the plasma membrane and cell wall accompanies a marked increase in the number and size of Golgi vesicles. Histochemical evidence using the thiocarbohydrazide-osmium vapor method shows polysaccharide to be present within Golgi vesicles while they are still attached to the Golgi apparatus. Mucilage deposition is associated with further cell expansion, separation of the protoplast from the cell wall, fusion of vacuoles and extra protoplasmic space, and the disappearance of starch.     

Consequences of the introduction of the planarian Girardia anceps (Tricladida: Dugesiidae) in artificial containers with larvae of the mosquitoes Aedes aegypti and Culex pipiens (Diptera: Culicidae) from Argentina

Aedes aegypti and Culex pipiens are container-dwelling mosquito species that are vectors of important diseases to man, such as dengue and lymphatic filariasis, respectively. Predators of these pests are an interesting alternative to be incorporated to biological control measures. We tested the consequences of introducing individuals of Girardia anceps, a native freshwater flatworm species, within artificial water containers where larvae of these mosquitoes thrive. Our goals were to ascertain if mosquito species, density of larvae (high or low), type of water container (tires or ovitraps), and presence or absence of planarians affected mosquito survivorship (measured as number of individuals reaching the pupa stage) in manipulated artificial containers. Furthermore, we monitored ovitraps in the field along several months in order to explore the long-term effect of the presence of planarian on the colonization of these containers by feral mosquitoes under natural conditions. We found that the presence of planarians reduced the number of mosquitoes reaching pupation and that such reduction depends on the initial density of larvae. Reduction of populations of A. aegypti was high along the breeding season of this mosquito, being the effect less evident in C. pipiens. G. anceps could be an agent of control against container-breeding mosquitoes if its release in small water containers is complemented with other suitable management strategies.     

Environmental characteristics of the cemeteries of Buenos Aires City (Argentina) and infestation levels of Aedes aegypti (Diptera: Culicidae)

Cemeteries with many water-filled containers, flowers, sources of human blood, and shade are favorable urban habitats for the proliferation of Aedes aegypti, a vector of yellow fever and dengue. A total of 22,956 containers was examined in the five cemeteries of the city of Buenos Aires, Argentina. The vector was found in four cemeteries that showed an average infestation level of 5.5% (617 positive out of 11,196 water-filled containers). The four cemeteries positive for Ae. aegypti showed significantly different (p<0.01) infestation levels. Vegetation cover and percentage of infestation were significantly correlated (p<0.01), but neither cemetery area nor number of available containers were significantly related to the proportion of positive vases. Our results suggest that the cemeteries of Buenos Aires represent a gradient of habitat favorableness for this vector species, some of which may act as foci for its proliferation and dispersal.     

A rapid micromethod for the determination of nitrogen and phosphate in biological material

Two types of small containers for the liquid scintillation counting of 0.15–2 ml of aqueous sample in emulsifying agents have been investigated. Counting efficiency and reproducibility were found to be comparable with those found with more orthodox methods, but the effective cost is less. The systems also provide an economical way of using expensive reagents.     

How does surrounding vegetation affect the course of succession: A five‐year container experiment

Question: How does location and time of insertion affect the course of succession in experimental containers? Location: Benešov nad Lipou, Českomoravská vrchovina (Czech-Moravian uplands), Czech Republic Methods: We designed a 5-year container experiment in which plant succession started from scratch. Soil conditions were constant and all containers were filled with homogeneous substrate containing no propagules. We placed the containers in two contrasting habitats (meadow and floodplain) under identical climatic conditions but differing in surrounding vegetations and hence seed input. New containers were installed (and hence succession started) in two subsequent years, twice in each year (spring and autumn). We assume that the individual dates would lead to differences in propagule input and weather conditions. Results: Although both year and season of succession initiation considerably affected the initial species composition, we observed a pronounced convergence within the set of containers located in each habitat. However, the similarity of containers initiated at the same time but located in different habitats decreased over the course of succession. Final composition of the meadow and floodplain containers was therefore mostly determined by permanent seed input from their nearby neighborhood. Conclusions: This study demonstrated that propagule availability is an important determinant of the course of succession, and that differential seed input leads to different pathways of succession, even when all other environmental conditions are equal.     

HCMV在基因转染细胞中复制的研究

用人喉上皮细胞癌细胞系(HEP_2)的DNA转染人胚肺细胞(HEL),得到了5株基因转染细胞(GTC),命名为A5、B3、G8、D3和H3.这些GTC的染色体数在HEL的染色体数与两个亲代细胞(HEP-2和HEL)染色体的和数之间,感染人巨细胞病毒ADI69株后4d,D3比A5、B3、G8和H3有更多的荧光阳性细胞和更高的病毒滴度.在D3中HCMV复制随感染剂量的增大而增速.不同代数的D3对HCMV具有相似的敏感性,HCMV感染传至100代的D3,电镜证实仍可象原代D3复制大量的HCMV.永生性的D3可以用作分析调控HCMV复制的宿主细胞因子、分离培养HCMV等的工具.     

OBSERVATIONS ON THE ASSOCIATION OF HELICAL POLYRIBOSOMES AND FILAMENTS WITH VINCRISTINE-INDUCED CRYSTALS IN EARLE'S L-CELL FIBROBLASTS

Earle's L-929 fibroblasts from cultures treated with 5–10 µg/ml of vincristine sulfate have a large number of eosinophilic, proteinaceous crystals in their cytoplasm. In electron micrographs, large arrays of helical polyribosomes, stacks of Golgi lamellae, and membranes of granular endoplasmic reticulum are seen in the cytoplasm of these cells. "Stalks" of fine granular material, approximately 300 A wide, are often seen in association with the arrays of the helical polyribosomes. In many sections rows of helical polyribosomes and filaments emerging from individual polyribosomes are seen in intimate contact with the crystals. A gradual reduction in the number of crystals and crystal-bearing cells is seen in cultures removed from the drug-containing medium and reincubated in fresh medium. In electron micrographs, these reincubated cells show large aggregates of filamentous material in the cytoplasm, and in many sections filaments are seen in continuity with the crystals.