A new hypothesis on cell provisioning in solitary wasps

Like many other eumenid wasps, Pachodynerus nasidens (Latr.) generally stores more provisions in female–producing cells than in male–producing cells. According to the proposed hypothesis the provisioning of more or less food is determined by the variation in the time available for storing; this in turn, is caused by the slower or more rapid development respectively of female or male eggs.     

Pedilanthus diazlunanus (Euphorbiaceae): pollination by hymenopterans in a bird-pollinated genus

Pollination in the genus Pedilanthus is commonly effected by hummingbirds. Pollination by vespid wasps in Pedilanthus is documented for the first time based on observations of Pachodynerus nasidens and Eumenes americanus pollinating Pedilanthus diazlunanus. An hypothesis concerning the mechanism by which hummingbird pollination was replaced by insect pollination in Pedilanthus is advanced based on observations of insects on P. bracteatus.     

The biology of the eumenid mud-wasp Pachodynerus msidens in trapnests

Abstract. 1. The biology of the eumenid mud-wasp Pachodynems nasidens (Latr.) was studied in trapnests.
2. Females nested in traps placed in the shade and some nested in the same traps from which they had emerged.
3. The nesting behaviour and nest structure were generally similar to those of other eumenids that nest in borings, but a temporary crescent-shaped plug at the nest entrance and the use of both dry soil and mud for nest-building seems unique to P.nasidens .
4. Like most other eumenids, P.nasidens collected caterpillars, mainly Pyral-idae, Olethreutidae, Alucitidae and Thyrididae.
5. In linear nests, the innermost cells contained females and the outermost cells males, resulting in proterandry. One female can make several nests, each showing proterandry.
6. Female immatures were larger and took more time to develop than males. Also, their cells were larger and stocked with more food than male cells, hence needing more time to be provisioned.
7. The greater 'cost' to produce a female and a sex ratio biased toward females in short traps and about unity in longer traps, leads to a comment on parental investment.     

Immature stages of the bee fly Ligyra satyrus (F.) (Diptera: Bombyliidae): A hyperparasitoid of canegrubs (Coleoptera: Scarabaeidae)

We describe the third- and fourth-instar larva, pupa and biology of the hyperparasitoid bee fly Ligyra satyrus (Fabricius). The larval and pupal morphology of the bee fly is typical for members of the subfamily Anthracinae. The bee fly larvae are found inside cocoons of the scoliid Campsomeris tasmaniensis Saussure, an external parasite of canegrubs (Coleoptera: Scarabaeidae) in sugarcane fields at Ayr and Bundaberg, Queensland. Ligyra satyrus has been recorded attacking only C. tasmaniensis . Ligyra and its relatives are all parasites of predatory and parasitic, ground-nesting aculeate Hymenoptera such as Sphecidae, Pompilidae, Tiphiidae and Scoliidae. Larval morphology of the hyperparasite is similar to other ectoparasitic Bombyliidae and pupal morphology is compared to that of species in the same and related subfamilies. Rates of hyperparasitism at Gordonvale reported in the early 1900s are relatively high, but our results show that currently they are very low at Ayr and Bundaberg. These results suggest that the impact of the bee fly on natural control of the canegrubs by scoliids varies considerably.     

Non-host plant extracts reduce oviposition of Plutella xylostella (Lepidoptera: Plutellidae) and enhance parasitism by its parasitoid Cotesia plutellae (Hymenoptera: Braconidae)

Botanical preparations, usually from non-host plants, can be used to manipulate the behaviour of insect pests and their natural enemies. In this study, the effects of extracts of Chrysanthemum morifolium, a non-host plant of the diamondback moth, Plutella xylostella (Linnaeus), on the olfactory and oviposition responses of this phytophagous insect and on levels of parasitism by its specialist parasitoid Cotesia plutellae (Kurdjumov) were examined, using Chinese cabbage Brassica campestris L. ssp. pekinensis as the test host plant. Olfactometer tests showed that volatiles of chrysanthemum extract-treated host plants were less attractive to P. xylostella females than those from untreated host plants; and in contrast, volatiles of the chrysanthemum extract-treated host plants were more attractive to females of its parasitoid C. plutellae than those from untreated host plants. Oviposition preference tests showed that P. xylostella females laid only a small proportion of their eggs on chrysanthemum extract-treated host plants, while ovipositing parasitoid females parasitized a much higher proportion of host larvae feeding on the treated host plants than on untreated host plants. These results suggest that certain non-host plant compounds, when applied onto a host plant, may render the plant less attractive to a phytophagous insect but more attractive to its parasitoids. Application of such non-host plant compounds can be explored to develop push-pull systems to reduce oviposition by a pest insect and at the same time enhance parasitism by its parasitoids in crops.     

Preirradiation of host (monkey) cells mitigates the effects of UV upon simian virus 40 DNA replication

We are examining the effects of preirradiation of host (monkey) cells upon the replication of UV-damaged SV40. Control cells and cells preirradiated with low fluences (5 or 10 J/m2) of UV were infected with undamaged SV40, and the immediate effects of a subsequent irradiation were determined. UV inhibited total SV 40 DNA synthesis (incorporation of thymidine into viral DNA) in both preirradiated and control cells, but the extent of inhibition was less in the preirradiated cells. A test fluence of 60 J/m2 to SV40 replicating in preirradiated cells reduced synthesis only as much as a test fluence of 25 J/m2 in control cells. The fraction of recently replicated SV40 molecules that re-entered the replication pool and subsequently completed one round of replication in the first 2 h after UV was also decreased less in the preirradiated cells. Thus preirradiation of the host cell mitigates the immediate inhibitory effects of a subsequent UV exposure upon SV40 replication.     

Evidence for Exploitative Competition: Comparative Foraging Behavior and Roosting Ecology of Short-Tailed Fruit Bats (Phyllostomidae)

Chestnut short-tailed bats, Carollia castanea , and Seba's short-tailed bats, C. perspicillata (Phyllostomidae), were radio-tracked ( N = 1593 positions) in lowland rain forest at Tiputini Biodiversity Station, Orellana Province, Ecuador. For 11 C . castanea , mean home range was 6.8 ± 2.2 ha, mean core-use area was 1.7 ± 0.8 ha, and mean long axis across home range was 438 ± 106 m. For three C . perspicillata , mean home range was 5.5 ± 1.7 ha, mean core-use area was 1.3 ± 0.6 ha, and mean long axis was 493 ± 172 m. Groups of less than five C. castanea occupied day-roosts in earthen cavities that undercut banks the Tiputini River. Carollia perspicillata used tree hollows and buildings as day-roosts. Interspecific and intraspecific overlap among short-tailed bats occurred in core-use areas associated with clumps of fruiting Piper hispidum (peppers ) and Cecropia sciadophylla . Piper hispidum seeds were present in 80 percent of the fecal samples from C . castanea and 56 percent of samples from C . perspicillata . Carollia perspicillata handled pepper fruits significantly faster than C . castanea ; however, C . castanea commenced foraging before C . perspicillata emerged from day-roosts. Evidence for exploitative competition between C . castanea and C . perspicillata is suggested by our observations that 95 percent of ripe P . hispidum fruits available at sunset disappear before sunrise ( N = 74 marked fruits). Piper hispidum plants produced zero to 12 ripe infructescences per plant each night during peak production. Few ripe infructescences of P . hispidum were available during the dry season; however, ripe infructescences of C . sciadophylla , remained abundant.     

Resource utilization by the freshwater deposit feeder Ptychoptera townesi (Diptera: Ptychopteridae)

SUMMARY 1. False crane fly larvae, Ptychoptera townesi (Diptera), occurred in high densities in a flow-controlled section of stream where fine particulate organic matter (FPOM; 0.45 μm to 1 mm in diameter) had accumulated, but were quite rare both upstream and downstream from the section.
2. In laboratory studies, P. townesi grew only on FPOM less than 250 μm. Larvae consistently grew fastest when fed small particles (0.45–53 μm in diameter).
3. Ptychoptera townesi consumed relatively small amounts (0.002 mg per mg animal dry mass day⁻¹) of FPOM (0.45–53 μm). They had long gut content passage times (greater than 19 h) and relatively high efficiencies of conversion of ingested food to body substance (20.7%). Gut content passage times were variable, and depended partially on the nature of the substrate.
4. False crane fly larvae compacted FPOM into faecal pellets considerably larger in size than particles ingested. They lost mass when allowed to feed on their own faecal material, as well as on faeces greater than 250 μm in diameter produced by shredders. However, they survived and grew on shredder faeces (53–500 μm in diameter) that contained a mixture of smaller particles and particles too large for ingestion.
5. The overall pattern of resource utilization by P. townes involved slow handling of relatively small volumes of food, which probably passed once only through a complex alimentary tract.     

Effects of light intensity and addition of carotene rich Dunaliella salina live cells on growth and antioxidant activity of Solea senegalensis Kaup (1858) larval and metamorphic stages

Senegal sole Solea senegalensis larval and metamorphic stages were exposed to a range of light intensities (200, 1000 and 2000 lx) in cultures with or without supplementation of β-carotene-rich live Dunaliella salina cells. Antioxidant biomarkers such as superoxide dismutase (SOD), catalase (KAT), total glutathione peroxidase (t-GPX) and malondialdehyde (MDA) were determined in larval and metamorphic stages. Growth was not affected ( P > 0·05) either by light intensity or D. salina supplementation. Survival after metamorphosis was also unaffected by D. salina supplementation (mean ± s . e . 81·0 ± 2·5% against 80·6 ± 2·9% those fed the control algal diet) or light intensity (mean ± s . e . 74·3 ± 4·9% for 200 lx, 85·1 ± 2·7% for 1000 lx and 82·8 ± 5·2% for 2000 lx, respectively). Light intensity affected ( P < 0·05) KAT and t-GPX throughout development. SOD was only affected in metamorphosing larvae. The highest KAT and t-GPX activities were detected when the lowest light intensity (200 lx) was used. Light had no effect ( P > 0·05) on MDA at any stage. Supplementing the diet with D. salina did not affect SOD, KAT or t-GPX and there was no interaction ( P > 0·05) with light intensity. MDA was the only biomarker whose activity was significantly ( P < 0·05) reduced when D. salina was supplemented to the larval rearing tanks. The effect of D. salina supplementation was only detected in metamorphosing larvae, whose MDA levels were noticeably higher than in earlier stages. These results are evidence of the antiperoxidative effect of β-carotene from live algae in the larval rearing process of marine fishes.     

Isospora elmahalensis n. sp. (Apicomplexa, Eimeriidae), a Parasite of the White-Cheeked Bulbul (Pycnonotus leucogenys) in Saudi Arabia

Isospora elmahalensis n. sp. is described from the Saudi Arabian bird, Pycnonotus leucogenys , from the Elmahala valley. Sporulated oocysts of I. elmahalensis were spherical or nearly subspherical, 19.5–22.5 × 18.5–20 (21.34 ± 0.4 × 19.06 ± 0.5) μm. Oocysts lacked a micropyle, residuum, and polar granule. Sporocysts were ovoid, 14–17.5 × 7–12 (16.08 ± 1.05 × 9.9 ± 1.55) μm, and had a Stieda body and sporocyst residuum, but lacked a substiedal body. Sporozoites were elongated with a clear globule at one end. The host bird belongs to the order Passeriformes.     

Kinetics of deoxyribonucleic acid destruction and synthesis during growth of Bdellovibrio bacteriovorus strain 109D on pseudomonas putida and escherichia coli

During the growth of Bdellovibrio bacteriovorus on Pseudomonas putida or Escherichia coli in either 10(-3)m tris(hydroxymethyl)aminomethane or in dilute nutrient broth, the host deoxyribonucleic acid (DNA) was rapidly degraded, and by 30 to 60 min after the initiation of the bdellovibrio development cycle essentially all host DNA became nonbandable in CsCl gradients. At this stage the host DNA degradation products were nondiffusable, and there was no appreciable pool of low-molecular-weight (cold acid soluble) DNA fragments in the cells or in the suspending medium. Bdellovibrio DNA synthesis occurred only after degradation of host DNA to a nonbandable form was complete. The synthesis occurred in a continuous fashion with P. putida as the host and in two separate periods with E. coli as host. By using E. coli containing a (3)H-thymidine label, it was shown that 73%, on the average, of the thymine residues of host DNA were incorporated into bdellovibrio DNA when E. coli was the only source of nutrient. In the presence of dilute nutrient broth, the host cells still served as the major source of precursors for bdellovibrio DNA synthesis, with only 20% of the precursors arising from the exogenous nutrients. The data indicate an efficient and controlled utilization of host DNA by the bdellovibrio. The host DNA is apparently degraded early in the developmental cycle to oligonucleotides of intermediate molecular weight from which the biosynthetic monomers are generated only as they become needed for bdellovibrio DNA synthesis.     

Space use of the spotted sand lizard (Pedioplanis l. lineoocellata) under different degradation states

Although the effects of grazing-induced savannah degradation on animal diversity are well documented, knowledge of how they affect space use or responding behaviour remains poor. In this study, we analysed space use of the spotted sand lizard ( Pedioplanis l. lineoocellata ) in degraded versus nondegraded habitats of southern Kalahari savannah habitats. Lizards were radio tracked, daily movement distances recorded and home range sizes calculated. In degraded Kalahari savannah habitats where plant diversity and perennial grass cover are low but shrub cover high, P. lineoocellata moves larger distances (40.88 ± 6.42 m versus 27.43 ± 5.08 m) and occupies larger home ranges (646.64 ± 244.84 m² versus 209.15 ± 109.84 m²) than in nondegraded habitats (high plant diversity, high perennial grass cover and low shrub cover). We assume that this increase in daily movement distances and home range sizes is a behavioural plasticity to limited food resources in degraded savannah habitats. Although P. lineoocellata is able to adjust to resource-poor savannah habitats, the increase in the lizard's movement activities is likely to result in a higher predation risk. This is supported by the lower availability of protective vegetation i.e. perennial grass cover. Hence, we conclude that despite behavioural plasticity of P. lineoocellata , overgrazing has a severe negative impact on the space use of P. lineoocellata .     

Dehydroepiandrosterone and a beta-agonist, energy transducers, alter antioxidant enzyme systems: influence of chronic training and acute exercise in rats

We examined the influence of dehydroepiandrosterone (DHEA), a beta-agonist, and exercise training on enzymes that detoxify toxic oxygen species. Feeding 0.4% DHEA decreased hepatic cytosolic (c) selenium-dependent glutathione peroxidase (GPX), (-26%, P less than 0.0001) and increased hepatic mitochondrial (m) Mn superoxide dismutase (SOD), (+38%, P less than 0.001). DHEA decreased myocardial c-GPX (-21%, P less than 0.05) when compared to a beta-agonist (beta A; L644969 Merck and Co.) fed at 5 ppm but neither differed from the Control (C). In contrast, the beta A increased hepatic m-GPX (+25%, P less than 0.05). In skeletal muscle, DHEA and beta A decreased muscle c-GPX by 20 and 12%, respectively (P less than 0.0009). DHEA increased both muscle (+20%, P less than 0.01) and myocardial (+20%, P less than 0.05) c-glutathione S-transferase (GST) over beta A (+20%, P less than 0.01) but neither was significantly different from C. Similar to DHEA, chronic training (Tr) (1 h/day, 5 days/week at 27 m/min, 15% grade on treadmill) decreased hepatic c-GPX (-16%, P less than 0.003). Tr elevates muscle c-GPX (+36%, P less than 0.05) in C. Tr increased myocardial c-GPX by 28% in the beta A-treated rats, whereas Tr decreased myocardial c-GPX by 22% in the C (P less than 0.05, interaction). One hour of acute exercise (Ex) (70% VO2 max relative work load) decreased hepatic homogenate catalase (-12%, P less than 0.02) and increased hepatic m-Mn SOD (+28%, P less than 0.03). Ex decreased myocardial c-GST (P less than 0.05) only in the DHEA-treated rats. DHEA and Tr may improve efficiency of oxygen utilization at the tissue level with lower antioxidant enzyme activity in liver and locally protective up-regulation in muscle. beta A stresses oxygen utilization systems and liver responds by up-regulation of antioxidant enzymes. The increase in myocardial c-GPX activity in the beta A-treated group may be a protective effect against indirect catecholamine-induced myocardial necrosis which results from free radical generation.     

白细胞介素4降低脐带间充质干细胞对造血干细胞分化潜能的维持能力

目的探讨白细胞介素4（IL-4）对脐带间充质干细胞（UC-MSC）维持造血干细胞分化的影响。 方法将UC-MSC与脐带血CD34⁺造血干细胞按照造血支持能力常用的方案共培养,实验分为对照组和IL-4组,IL-4处理组加入IL-4（20 ng/ml）培养14 d。收集细胞并计数,使用流式细胞仪检测表达CD34的细胞比例。取3×10³个细胞,加入到半固体培养基,培养14 d后,通过倒置显微镜观察比较各种集落的形成,并使用流式细胞仪分析其中巨噬细胞和粒细胞表面特异性蛋白CD11b、CD14和CD15的表达。对两独立样本进行t检验统计学分析。 结果加入IL-4后,共培养体系中细胞数量（1.31±0.05）×10⁵个/孔与对照组（2.80±0.28）×10^5?个/孔相比下降,差异有统计学意义（t = 7.31,P < 0.05）,并且流式细胞分析显示其中的CD34⁺细胞比例也有降低。3×10³个IL-4组得到的细胞形成巨噬细胞集落形成单位的能力（9.33±1.53）?个/孔较对照组（17.67±0.58）个/孔有明显下降,差异有统计学意义（t = 8.84,P?< 0.001）;形成粒-巨噬集落形成单位的能力（15.67±3.22）个/孔较对照组（29.33±4.04）?个/?孔有明显下降,差异有统计学意义（t = 4.58,P < 0.05）;形成总集落单位的能力（39.33±9.07）个/?孔较对照组（62.67±6.66）?个/?孔也有下降,差异有统计学意义（t = 3.59,P < 0.05）。IL-4组得到的细胞分化出的细胞总数（3.67±1.71）×10⁵个/孔与对照组（9.50± 3.13）×10⁵个/孔相比也明显下降,差异有统计学意义（t = 2.83,P < 0.05）,而流式细胞术分析发现分化成的细胞中CD14⁺细胞比例也下降。 结论IL-4可以降低UC-MSC对造血干细胞分化潜能的维持能力,提示在Ⅱ型辅助T细胞相关体液免疫疾病中使用间充质干细胞治疗时,也需要兼顾机体造血相关功能。     

温度对产虫茶昆虫紫斑谷螟生长发育的影响

为探明温度对贵州主要产虫茶昆虫紫斑谷螟Pyralis farinalis (Linnaeus)生长发育的影响, 本研究以白茶Litsea coreana为寄主植物, 分别设置5个恒温（19, 22, 25, 28和31℃）条件, 研究温度对紫斑谷螟卵、 幼虫、 蛹和未成熟期平均发育历期、 发育速率和存活率的影响, 计算各虫态发育起点温度和有效积温。 结果表明： 温度对紫斑谷螟各虫态发育历期、 发育速率和存活率影响显著。 在19 ~ 31℃范围内, 各虫态的平均发育历期均随温度的升高而缩短, 卵期、 幼虫期、 蛹期及未成熟期均在31℃达到最小值, 分别为4.56±0.24, 43.33±1.50, 7.89±0.20和55.78±1.69 d。 紫斑谷螟各虫态发育速率与温度呈二次回归关系, 且极显著相关。 此外, 温度显著影响各虫态存活率, 卵的存活率在28℃时最高, 为93%; 幼虫和蛹的存活率则在25℃最高, 分别为88%和93%; 温度过高或过低均不利于其生长发育。 由直接最优法计算得到紫斑谷螟卵期、 幼虫期、 蛹期及未成熟期的发育起点温度分别为13.30, 15.48, 13.19和14.82℃, 有效积温依次为88.36, 679.51, 159.73和952.04日·度。 这些结果为紫斑谷螟的繁殖提供了基础参考数据, 对指导虫茶生产有实用参考价值。     

Comparison of several techniques for mobility and activity estimates of smallmouth bass in lentic environments

Mean daily mobility estimates for smallmouth bass Micropterus dolomieu from mark-recapture (47·5±12·5 m) were less than from conventional telemetry (77·1±10·6 m). The relationship developed in a respirometer between the activity transmitter and swimming speed ( r 2=0·99, P < 0·001, n =6) when applied to field activity data estimated mean daily movement at 27 408±4085 m, i.e. >100 times mark/recapture or telemetry estimates. Using these estimates in the activity parameter of a bioenergetics model resulted in different model outputs. These results highlight the potential underestimates in activity associated with using traditional mark-recapture and locational telemetry and reaffirm that fish expend a significant portion of their activity budgets undertaking localized movements.     

Proteomic comparison of membrane and extracellular proteins from invasive (PAO1) and cytotoxic (6206) strains of Pseudomonas aeruginosa

Strains of Pseudomonas aeruginosa can be phenotypically classified by their mode of pathogenicity as either invasive, where the bacterium is internalised by host cells, or cytotoxic, where the host cell is killed without internalisation through the expression of cytotoxicity factors. These phenotypes are thought to depend primarily on the interactions of pseudomonal membrane and secreted proteins with host cells. We report here comparisons of outer membrane and extracellular protein-enriched fractions from invasive (PAO1) and cytotoxic (6206) strains of P. aeruginosa separated by two-dimensional (2-D) gel electrophoresis. Gel image comparisons revealed the two strains express essentially identical membrane protein profiles under the conditions investigated. Membrane protein strain differences were typically the result of minor amino acid sequence variations resulting in small mass and isoelectric point shifts visible on 2-D gels. Analysis of extracellular proteins from stationary phase growth, however, revealed significantly different protein profiles. Extracellular fractions from the invasive PAO1 strain were dominated by extracellular proteases including elastase (LasB), LasA protease and chitin-binding protein, as well as several previously designated 'hypothetical' proteins. LasB appeared to be highly processed with 28 discrete mass and isoelectric point forms detected in this study. The significant number of active extracellular proteases (including LasB itself) may account for this processing. Conversely, extracellular fractions from strain 6206 consisted mainly of cellular and membrane exposed proteins including GroEL, DnaK and flagellar subunits. These are thought to result from cellular turnover during growth and the reliance on the secretory mechanisms of this strain to produce high levels of cytotoxicity factors, such as ExoU, which may be produced only upon specific interactions with host cells. These studies will aid in elucidating the differences between invasive and cytotoxic P. aeruginosa at the proteome level.     

A molecular phylogeny for digger wasps in the tribe Ammophilini (Hymenoptera,Apoidea, Sphecidae)

The evolution of parental care strategies in aculeate (stinging) wasps and bees has been much studied from a functional perspective, but relatively little phylogenetic information is available to place this in a rigorous historical context, especially at the species level. We used mitochondrial cytochrome oxidase I and two nuclear genes, the elongation factor‐1α and LW rhodopsin, to investigate the phylogeny of Sphecidae digger wasps. We focus particularly on the tribe Ammophilini, a clade of nonsocial apoid wasps that exhibit unusually diverse parental care strategies. We analysed a 2232 bp dataset for 40 ammophilines plus nine other taxa from within the remaining Sphecidae. Our Bayesian phylogeny provides strong support for the monophyly of Ammophilini and for the monophyly of all six individual ammophiline genera, except that the position of P. affinis within the genus Podalonia is only weakly supported. The monophyly of some, but not all, previously designated species groups within the genus Ammophila is supported. We discuss the implications of our results for the evolution of morphological traits used previously in ammophiline systematics.     

The effect of buffer osmolality on the survival of cat (Felis catus ) spermatozoa at 5 degrees C

Cat semen was diluted at 37 degrees C in Tes-Tris buffer (TesT), pH 7.5, at osmolalities ranging from 195 to 390 m0sm/kg, cooled to 5 degrees C over 90 minutes and stored for 24 hours at that temperature. Motility and percentage of spermatozoa staining with a supravital stain were estimated before cooling, after cooling and after storage for 24 hours. The osmolality of undiluted pooled ejaculates from five animals was measured, and also that of different diluents (citrate with phosphate buffer, lactose and TesT-egg yolk) used for cat semen. The osmolality measurements of cat semen suggested an osmolality of less than 320 m0sm/kg at ejaculation, increasing with time after ejaculation. Varying the egg yolk concentrations (2% to 20%) did not affect the osmolality of TesT diluent. Diluent osmolalities of less than 292 m0sm/kg were found to reduce sperm motility significantly (P <0.001 ) although there was no significant increase in the percentage of cells staining with a supravital stain, while those greater than 325 m0sm/kg increased the variation of response among animals. Cooling and storage significantly reduced motility (P <0.01 to P <0.001 ) and increased the number of stained cells (P <0.001 ). There were significant differences between ejaculates (P <0.01 ) and significant interactions between osmolality and cooling/storage (P <0.05 to P <0.001 ). The best overall results were seen with a TesT diluent of 292 to 325 m0sm/kg which supported good motility for at least 24 hours.