Resistance of Apanteles eggs to the haemocytic encapsulation by their habitual host, Pieris

The calyx fluid in the lateral oviduct of a gregarious parasitoid, Apanteles glomeratus contained ellipsoid particles of ca. 130 × 200 nm. These calyx fluid particles did not appear to be embedded in a fibrous outer layer on the surface of eggs in the lateral oviduct. They were not observed on the surfaces of the eggs 3 to 4 hr after being deposited into the host haemocoele. Oviposition experiments indicated that the occurrence of haemocytic defence reactions of the late 2nd instar larvae of the Pieris rapae crucivora against 1 st instar larvae of the parasitoid increased with a decreasing number of the parasitoid eggs introduced into a host, and that more than 5 to 9 parasitoid eggs were needed for suppressing the ability of the host to encapsulate its parasitoid larvae immediately after hatching. When eggs with calyx fluid obtained from egg reservoir were injected into the host, they were found to be encapsulated 1 to 2 days after the injection. They could not start their embryonic development. When calyx fluid-free 3-hr-old eggs were injected in a number of more than 5 eggs into a 5th instar larva of Pieris, 58% of 31 eggs injected had normally hatched without evoking encapsulation reactions by the host. Both electron microscopic observations of parasitoid eggs in the host haemocoele and the experimental results suggested that calyx fluid or calyx fluid particles of the parasitoid might not be involved in the encapsulation-inhibiting activity of the parasitoid eggs. Rather it was anticipated that a substance (or substances) might be secreted by the parasitoid eggs into the haemocoele of the host, which suppressed defence reactions of the host.     

雷氏黄萤幼虫呼吸系统和呼吸行为

研究雷氏黄萤Luciola leii Fu and Ballantyne幼虫的呼吸系统及其呼吸行为。结果表明:雷氏黄萤幼虫的呼吸系统中只有气管无气囊。前胸、中胸和后胸均分布有气门,无气管鳃,腹部1~8节分布有气门和气管鳃,气门腔基部和气管鳃基部相连,呈"√"状,气管鳃内气管与气门气管相连通。雷氏黄萤幼虫的呼吸行为分为3种:利用胸部气门呼吸、腹部气门呼吸和气管鳃呼吸,其中以腹部气门呼吸为主。     

小鼠眼神经注入Dil后三叉神经节的形态学研究

目的：经眼神经注入DiI研究小鼠三叉神经节的形态学结构。方法：小鼠10只,体重25—30克,雌雄不拘,进行灌注固定后,在外科显微镜下开颅并确认三叉神经节和眼神经,分别于双侧眼神经植入DiI染色晶体。37℃恒温箱放置3个月,待DiI染色晶体扩散后,取出植入DiI染色晶体的眼神经和三叉神经节,再根据神经走向切片,通过荧光显微镜观察DiI染色晶体在三又神经节内的分布。结果：眼神经离三叉神经节约1cm处植入DiI染色晶体后,应用荧光显微镜明视野观察,均可见到高密度标记的眼神经纤维,行向后内,穿经眶上裂入颅。逐步靠近三叉神经节外上方,并进入三叉神经节内,眼神经标记的神经元位于三叉神经节的前内侧。在三叉神经节内可见到DiI标记的神经节细胞及神经纤维。神经纤维平行致密排列,并被神经节细胞神经纤维分隔成群或簇。神经节细胞呈圆形和卵圆形,大小不一,部分节细胞呈蜂窝状排列。亦可见神经元的突起,有的呈螺旋状连于胞体,有的呈线状连于胞体,并可见到双极神经元。结论：小鼠经眼神经注入DiI后,三叉神经节细胞和神经纤维的排列循序跟其他动物基本一致。     

普通齿蛉幼虫的呼吸系统及呼吸行为

研究了普通齿蛉Neoneuromus ignobilis Navás幼虫的呼吸系统及其呼吸行为。结果表明:普通齿蛉幼虫为全气门式(10对气门)呼吸系统,前中胸、中后胸之间、腹部8节各有1对气门,腹部8节各有气管鳃1对,前6对细短,管状,有较短绒毛,后2对气管鳃较粗长,呈羽毛状。腹部1～7节各有1对毛簇,第8腹节无毛簇。侧纵干气管较粗,4束,自前胸前缘部分成左右2组,每组两根侧纵干气管,向胸腹部延伸,二级气管分别伸达各个气门和毛簇,腹部每节由毛簇处的二级气管分支而来的三级气管相连或延伸至消化道等处。气管鳃中无气管。有毛簇呼吸、气门呼吸和体壁呼吸3种呼吸方式,在水中以毛簇呼吸为主,在陆上进行气门呼吸和体壁呼吸。     

Observations on cellular immunity and parasitism in the tussock moth

Cellular responses to the introduction of foreign objects into the haemocoele of both control and parasitized tussock moth larvae were examined. In normal larvae, the response to large foreign objects such as Hyposoter fugitivus eggs and Sephadex beads was encapsulation, accompanied by a rapid and sustained increase in the total haemocyte count. Smaller objects such as yeast cells were cleared into nodules within a matter of minutes; nodulation too was accompanied by an increased total haemocyte count.In larvae parasitized by the braconid wasp Cotesia melanoscela, both encapsulation and nodulation were permanently suppressed. Inhibition of these normal cellular defence reactions was accompanied by a reduction in the total haemocyte count, the appearance of debris in the haemocoele, and by nuclear pycnosis in an unidentified population of cells; however, since extensive nuclear pycnosis also occurred in haemopoietic tissue, it is assumed that prohaemocytes may represent a target cell population. All of the observed effects required the presence of both C. melanoscela calyx fluid and venom in the host animal. Phagocytosis as an immune response remained essentially intact, and was capable of completely clearing both yeast and Escherichia coli cells injected into the haemocoele.     

Identification of multiple odorant receptors essential for pyrethrum repellency in Drosophila melanogaster

Pyrethrum extract from dry flowers of Tanacetum cinerariifolium (formally Chrysanthemum cinerariifolium) has been used globally as a popular insect repellent against arthropod pests for thousands of years. However, the mechanistic basis of pyrethrum repellency remains unknown. In this study, we found that pyrethrum spatially repels and activates olfactory responses in Drosophila melanogaster, a genetically tractable model insect, and the closely-related D. suzukii which is a serious invasive fruit crop pest. The discovery of spatial pyrethrum repellency and olfactory response to pyrethrum in D. melanogaster facilitated our identification of four odorant receptors, Or7a, Or42b, Or59b and Or98a that are responsive to pyrethrum. Further analysis showed that the first three Ors are activated by pyrethrins, the major insecticidal components in pyrethrum, whereas Or98a is activated by (E)-β-farnesene (EBF), a sesquiterpene and a minor component in pyrethrum. Importantly, knockout of Or7a, Or59b or Or98a individually abolished fly avoidance to pyrethrum, while knockout of Or42b had no effect, demonstrating that simultaneous activation of Or7a, Or59b and Or98a is required for pyrethrum repellency in D. melanogaster. Our study provides insights into the molecular basis of repellency of one of the most ancient and globally used insect repellents. Identification of pyrethrum-responsive Ors opens the door to develop new synthetic insect repellent mixtures that are highly effective and broad-spectrum.     

Effects of temperature and humidity on transpiration in adults of the lesser mealworm,Alphitobius diaperinus (Coleoptera: Tenebrionidae)

Measurements of water loss were made on adults of the lesser mealworm, Alphitobius diaperinus, using a recording micro-electrobalance and a programmable heat circulator bath. This species originates in tropical regions and infests poultry houses in temperate countries. Two routes of water loss were examined: the general cuticle and via the spiracles. Temperature and relative humidity of the ambient air substantially affect the cuticular transpiration in adults (fresh body weights from 12 mg to 22 mg). At near 0% R.H., between 20 and 40 degrees C the rate of body water loss gradually increased; on the other hand, the insects gained weight in an atmosphere close to saturation. Above 40 degrees C transpiration flow abruptly increased coinciding with the start of vigorous locomotor activity. This critical point corresponds to the opening of the spiracles from which the water is expelled from the tracheal system.In dead specimens, killed by cyanide or solvent, the water vapour slowly diffused out of the spiracles and, as in atracheate insects, the transpiration curves did not show a peak as the air temperature was increased.The thermostupor point (TSP) occurred as the insects became motionless; the corresponding temperature is significantly affected by atmospheric relative humidity (TSP=47.4+/-0.6 degrees C at c. 0% R.H.; TSP=46.6+/-0.7 degrees C at c. 100% R.H.).The transpiration flow was about four times as fast in specimens treated with solvent as in the individuals (live or cyanide-killed) that had undamaged water-proof cuticle. This species has to cope with a double challenge: (i) to adapt its physiology and ecology to poultry-house conditions which constitutes an extension of its primary habitats, and (ii) to survive over winter; high drought resistance and heat tolerance may constitute a pre-adaptation to conquer anthropogenic air-conditioned sites.     

Effect of inhibitors on chloride-dependent transmural potential in the rectal wall of Schistocerca gregaria

Previous experiments in vitro revealed a transmural potential difference (PD) and a short-circuit current (Isc) across the rectal wall of Schistocerca gregaria, which were dependent on chloride ions in lumen. The present report shows that anoxia, dinitrophenol and cyanide inhibit DP and Isc, proving that the required energy derives from oxidative metabolism. Acetazolamide also inhibits DP and Isc. Ouabain, when in haemocoele, has also an inhibitory effect, not ascribable to a Na⁺K⁺ pump blocking action. It is suggested that HCO₃^? plays an important rôle in the active transport of Cl^? from lumen to haemocoele and that ouabain may in some way inhibit chloride pumping.     

Pump out the volume—The effect of tracheal and subelytral pressure pulses on convective gas exchange in a dung beetle, Circellium bacchus (Fabricus)

Many flightless beetles like the large apterous dung beetle Circellium bacchus, possess a subelytral cavity (SEC) providing an extra air space below the elytra which connects to the tracheal system (TS) via metathoracic and abdominal spiracles. By measuring subelytral and intratracheal pressure as well as body movements and gas exchange simultaneously in a flow-through setup, we investigated the contribution of convection on Circellium respiratory gas exchange.No constriction phase was observed. TS and SEC pressures were always around atmospheric values. During interburst phase open abdominal spiracles and a leaky SEC led to small CO₂-peaks on a continuous CO₂ baseline, driven by intermittent positive tracheal pressure peaks in anti-phase with small negative subelytral pressure peaks caused by dorso-ventral tergite action.Spiracle opening was accompanied by two types of body movements. Higher frequency telescoping body movements at the beginning of opening resulted in high amplitude SEC and TS pressure peaks. High frequency tergite movements caused subelytral pressure peaks and led to a saw tooth like CO₂ release pattern in a burst. We propose that during the burst open mesothoracic spiracles increase the compliance of the subelytral cavity allowing big volumes of tracheal air being pulled out by convection.     

Expression and function of the empty spiracles gene in olfactory sense organ development of Drosophila melanogaster

In Drosophila, the cephalic gap gene empty spiracles plays key roles in embryonic patterning of the peripheral and central nervous system. During postembryonic development, it is involved in the development of central olfactory circuitry in the antennal lobe of the adult. However, its possible role in the postembryonic development of peripheral olfactory sense organs has not been investigated. Here, we show that empty spiracles acts in a subset of precursors that generate the olfactory sense organs of the adult antenna. All empty spiracles-expressing precursor cells co-express the proneural gene amos and the early patterning gene lozenge. Moreover, the expression of empty spiracles in these precursor cells is dependent on both amos and lozenge. Functional analysis reveals two distinct roles of empty spiracles in the development of olfactory sense organs. Genetic interaction studies in a lozenge-sensitized background uncover a requirement of empty spiracles in the formation of trichoid and basiconic olfactory sensilla. MARCM-based clonal mutant analysis reveals an additional role during axonal targeting of olfactory sensory neurons to glomeruli within the antennal lobe. Our findings on empty spiracles action in olfactory sense organ development complement previous studies that demonstrate its requirement in olfactory interneurons and, taken together with studies on the murine homologs of empty spiracles, suggest that conserved molecular genetic programs might be responsible for the formation of both peripheral and central olfactory circuitry in insects and mammals.     

Controlled mechanized trepan.

A mechanized trepan is described whose cutting action is halted automatically when the trepan's cutting edge reaches the last 0.03 mm. tissue layer of the cornea. The automatic half of both the motor which advances the trepan as well as the second motor which rotates the trepan is triggered by the sudden change in electrical resistance between the trepan and the patient's internal body fluid, at the final stage of penetration. This automatic feature eliminates the danger of inadvertant damage to the inner structures of the eye.     

条背萤幼虫不同发育阶段呼吸系统构造的观察和比较

通过显微解剖,结合透射电镜和扫描电镜,观察比较了条背萤Luciola substriata幼虫两个不同发育阶段呼吸系统的差异。结果表明：1～2龄幼虫的呼吸系统中只有气管无气囊,3～6龄幼虫的呼吸系统中气管和气囊并存。1～2龄幼虫的尾气门和3～6龄幼虫的腹部侧气门及尾气门结构没有差异。透射电镜观察提示条背萤1～2龄幼虫体壁上的毛状物为气管鳃。     

Effects of temperature and oxygen availability on water loss and carbon dioxide release in two sympatric saproxylic invertebrates

Water loss and VCO(2) relative to temperature and oxygen tension was investigated in a log-dwelling onychophoran (Euperipatoides rowelli) and a sympatric, un-described millipede species using flow-through respirometry. Onychophorans possess a tracheal system featuring permanently open spiracles. Total body water loss was consistently very high in E. rowelli and there was a positive correlation with increasing temperature. CO(2) output was continuous, increasing with higher temperatures and decreasing under lower oxygen concentrations. The millipede which has occludible spiracles also showed continuous gas exchange; however water loss was up to an order of magnitude lower than E. rowelli. An ability to survive under hypoxia is apparent for both species and corresponds with reports of hypoxic conditions within rotting logs. The rotting log habitat common to both taxa is characterized by high relative humidity and typically cool temperatures that approach 0 degrees C at night in winter. Consequently, dispersal through the higher temperatures and lower humidity of the exposed and dry understorey between suitable habitat may be hazardous for E. rowelli due to high desiccation susceptibility.     

Comparison of the cold hardiness capacities of the oviparous and viviparous forms of Lacerta vivipara

The lizard Lacerta vivipara has allopatric oviparous and viviparous populations. The cold hardiness strategy of L. vivipara has previously been studied in viviparous populations, but never in oviparous ones. The present study reveals that both the oviparous and viviparous individuals of this species are able to survive in a supercooled state at -3 degrees C for at least one week when kept on dry substrates. The mean crystallisation temperatures of the body, around -4 degrees C on dry substrata and -2 degrees C on wet substrata, do not differ between oviparous and viviparous individuals. All the individuals are able to tolerate up to 48-50% of their body fluid converted into ice, but only viviparous individuals were able to stabilize their body ice content at 48%, and hence were able to survive even when frozen at -3 degrees C for times of up 24 hours. Ice contents higher than 51% have been constantly found lethal for oviparous individuals. This suggests that, in L. vivipara, the evolution towards a higher degree of freezing tolerance could parallel the evolution of the viviparous reproductive mode, a feature believed to be strongly selected under cold climatic conditions. This is the first report, among reptiles, of an intraspecific variation regarding the freeze tolerance capacities.     

Studies on Trypanosoma rangeli Tejera, 1920. VI. Developmental pattern in the haemolymph of Rhodnius prolixus

The morphological sequence of Trypanosoma rangeli development in the body cavity of Rhodnius prolixus is described. The metacyclic trypanosome is the product of successive division and transformation during the intra and extracellular development in the haemocoele. The significance of the early invasion of T. rangeli into the haemolymph is discussed. The epidemiological importance of the developmental pattern of T. rangeli in the vectors haemolymph and the host-response to the parasite are considered.     

Lack of evidence in vivo for nitrergic inhibition by Escherichia coli (STa) enterotoxin of fluid absorption from rat proximal jejunum

Fluid absorption from the proximal jejunum of the anaesthetised rat was measured in vivo by fluid recovery. As expected, heat stable (STa) enterotoxin from E. coli reduced fluid absorption. Neither intraperitoneal L-NAME, thought to inhibit a putative neurally mediated action of STa, nor similar doses of D-NAME, ameliorated the inhibitory effect on jejunal fluid absorption of STa. Luminally perfused 10 mM sodium nitroprusside (SNP) had no effect on fluid absorption when expressed per gram dry weight per hour but reduced fluid absorption when expressed per cm length per hour. Similarly, 80 but not 40 mg/Kg of L-NAME reduced fluid absorption when expressed per cm length per hour, while the same dose of D-NAME did not. L-NAME and SNP significantly increased the wet weight to dry weight and the length to dry weight ratio of perfused loops. We conjecture that smooth muscle relaxation caused by these compounds increases interstitial fluid volumes that can be misconstrued as changes in absorption when this is expressed per cm length or per tissue wet weight. When fluid absorption is expressed per gram dry weight of tissue, there is no evidence for a role of nitric oxide in normal or STa inhibited fluid absorption.     

Chemical Diversity of the Natural Populations of Dalmatian Pyrethrum (Tanacetum cinerariifolium (Trevir.) Sch.Bip.) in Croatia

Dalmatian pyrethrum (Tanacetum cinerariifolium (Trevir. ) Sch.Bip. ) is a plant species endemic to the east Adriatic coast. The bioactive substance of Dalmatian pyrethrum is a natural insecticide, pyrethrin, a mixture of six active components (pyrethrins I and II, cinerins I and II, and jasmolins I and II). The insecticidal potential of pyrethrin was recognized decades ago, and dried and ground flowers have traditionally been used in Croatian agriculture and households. A total of 25 Dalmatian pyrethrum populations from Croatia were studied to determine the pyrethrin content and composition, and to identify distinct chemotypes. The total pyrethrin content ranged from 0.36 to 1.30% (dry flower weight; DW) and the pyrethrin I/pyrethrin II ratio ranged from 0.64 to 3.33%. The statistical analyses revealed that the correlations between the percentage of pyrethrin I and of all the other components were significant and negative. The total pyrethrin content was positively correlated with the percentage of pyrethrin I and negatively correlated with cinerin II. The multivariate analysis of the chemical variability enabled the identification of five chemotypes among 25 Dalmatian pyrethrum populations. The chemical characterization of indigenous Dalmatian pyrethrum populations may serve as a good background for future breeding and agricultural exploitation.     

Ovicidal and adulticidal activities of Cinnamomum zeylanicum bark essential oil compounds and related compounds against Pediculus humanus capitis (Anoplura: Pediculicidae)

The toxicity of cinnamon, Cinnamomum zeylanicum, bark essential oil compounds against eggs and adult females of human head louse, Pediculus humanus capitis, was examined using direct contact and vapour phase toxicity bioassays and compared with the lethal activity of their related compounds, benzyl alcohol, cinnamic acid, cinnamyl acetate, 4-hydroxybenzaldehyde and salicylaldehyde, as well as two widely used pediculicides, d-phenothrin and pyrethrum. In a filter-paper contact toxicity bioassay with female lice at 0.25 mg/cm², benzaldehyde was 29- and 27-fold more toxic than pyrethrum and d-phenothrin, respectively, as judged by median lethal time (LT₅₀) values. Salicylaldehyde was nine and eight times more active than pyrethrum and d-phenothrin, respectively. Pediculicidal activity of linalool was comparable with that of d-phenothrin and pyrethrum. Cinnamomum bark essential oil was slightly less effective than either d-phenothrin or pyrethrum. Benzyl alcohol and (E)-cinnamaldehyde exhibited moderate pediculicidal activity. After 24 h of exposure, no hatching was observed with 0.063 mg/cm² salicylaldehyde, 0.125 mg/cm² benzaldehyde, 0.5 mg/cm² Cinnamomum bark essential oil, 1.0 mg/cm² (E)-cinnamaldehyde, and 1.0 mg/cm² benzyl cinnamate. Little or no ovicidal activity was observed with d-phenothrin or pyrethrum. In vapour phase toxicity tests with female lice, benzaldehyde and salicylaldehyde were much more effective in closed containers than in open ones, indicating that the mode of delivery of these compounds was largely due to action in the vapour phase. Neither d-phenothrin nor pyrethrum exhibited fumigant toxicity. Cinnamomum bark essential oil and test compounds described merit further study as potential pediculicides or ovicides for the control of P. h. capitis.     

Age-related changes in the rate of water loss and survival time in dry air of active Drosophila melanogaster

By continuously monitoring the weight loss in dry air at 24.5°C of active individual male Drosophila melanogaster, it was shown that: (i) during the active period, weight loss is a linear function of time, but once the body water content is reduced to below a critical level, activity ceases and the rate of weight loss increases rapidly; the time at which the rate of weight loss increases probably coincides with the time of death; (ii) the sudden change in the rate of weight loss can be prevented by blocking the spiracles; (iii) the rate of weight loss in the active period is positively correlated with age; (iv) young flies can tolerate a greater reduction in total body water than old flies. It is suggested that the age-related decrease in survival time in dry air is a consequence of changes in the initial water content of the flies, in their rate of water loss, in their activity, and in their tolerance of low water content.