大劣按蚊吸血活动与交配受精的关系

大劣按蚊Anophelos dirus是我国及东南亚地区的重要传疟媒介。研究表明成蚊羽化后,在未喂血的情况下,可有较高的交配受精率。在同一蚊笼,吸血机会完全相同的条件下,大蚊笼受精雌蚊吸血率为59.88%,未受精雌蚊的吸血率为35.38%;小蚊笼受精雌蚊的吸血率为74.13%,未受精雌蚊吸血率仅为43.56%。正常交配雌蚊的吸血率为49.71%,未进行交配的处女雌蚊吸血率为28.42%。同为正常交配蚊群,喂血组雌蚊受精率55.85%,不喂血组的受精率为44.51%。初步认为大劣按蚊的吸血活动与交配受精有一定的关系。     

蜕皮甾酮对淡色库蚊Culex pipiens pallens卵巢发育诱导的研究

根据淡色库蚊卵巢卵泡的生长,对该种类雌蚊成虫期的卵巢发育及蜕皮甾酮对吸血前雌蚊卵巢发育的诱导作用进行了研究。结果证明:蜕皮甾酮不影响羽化后早期雌蚊的卵巢发育,但能引起羽化后一天至吸血前的雌蚊的卵巢生长并伴有卵黄沉积,而正常情况下这种现象仅在雌蚊吸血后才发生。卵巢提取物的聚丙烯酰胺凝胶电泳证明,蜕皮甾酮诱导发育的卵巢与正常吸血雌蚊的卵巢在沉积的卵黄蛋白方面是一致的。     

食蟹猴疟原虫B株感染对斯氏按蚊寿命的影响

本试验比较了食蟹猴疟原虫Ｂ株感染的（感染组）,吸正常猴血的（对照组）及未吸血的斯氏按蚊存活情况。三组总平均寿命分别为２０．７６±９．３５ｄ、２５．５１±９．００ｄ和２２．０４±６．９７ｄ。感染组比对照组总平均寿命缩短４．７５ｄ;对照组比未吸血组平均延长３．４７ｄ;两组均差异非常显著（Ｐ＜０．０１）。结果提示,感染食蟹猴疟原虫的和未吸血的斯氏按蚊寿命均受到影响。     

猪尾猴疟原虫（P.inui）广西株配子体感染性变化规律的实验研究

用P.inui广西株经蚊传和血传接种4只猴子,让大劣按蚊每6小时吸血感染1次,连续数周,以蚊胃感染情况判断配子体的感染性。结果发现:本虫株配子体发育成熟的需72n+K小时;配子体生理寿命约12小时;配子体的感染性具有每隔2天,在后半夜出现高峰的周期性变化;血中大环状体百分比高峰与蚊媒感染高峰一致。     

吸血、交配对大劣按蚊、斯氏按蚊雌蚊卵巢发育、产卵的影响

大劣按蚊(Anophelesdirus)是我国海南岛和东南亚山林地区重要的传疟媒介。本文主要就其吸血量多少、受精与否对雌蚊卵巢发育、产卵的影响进行的初步观察。材料与方法大劣按蚊为我室建立的自然交配繁殖种群。饲养条件见刘连珠等(1986)。卵巢发育的观察标准:饱血雌蚊:腹部膨隆,时可见雌蚊尾部排出淡色液体;半饱血雌蚊:腹部不膨隆,仅见部分血液或腹部可见部分糖水、部分血液。常规喂血,吸血后雌蚊按饱血、半饱血组分笼,24、48、72小时后分别解剖,检查受精囊受精与否,卵巢发育按Chri-Stophers分期进行观察。交配和空间大小对大劣按蚊、斯氏按…     

Wistar大鼠鼠龄、性别和脾脏对约氏疟原虫子孢子入侵和红外期发育的影

Wistar大鼠对约氏疟原虫Plasmodiumyoeliiyoelii子孢子较为敏感,是研究约氏疟原虫红外期较为理想的动物模型(王兴相等,1985)。但有关该动物的一些生理因素对子孢子入侵和红外期的发育影响所知甚少。本文用对比研究的方法观察了鼠龄、性别和脾脏对于孢子入侵和红外期发育的影响,以便更好地利用该鼠疟模型进行红外期生物学特性的研究。材料和方法约氏疟原虫BY265株每周血传一次,每5周蚊传一次,保种于昆明株小鼠体内。斯氏按蚊(An.stephensi)成蚊羽化后3天用于吸血感染,感染后置于24±1℃、RH80±10%温室内饲养,第14天按Pachecoetal(1979)…     

苯醚威作用于印鼠客蚤的组织学研究

【目的】通过研究苯醚威对印鼠客蚤 Xenopsylla cheopis (Rothschild,1903)的早 3 龄幼虫和未吸血新羽化成虫的组织学变化,探讨其灭蚤机理,为鼠疫媒介蚤种的防治提供基础资料。【方法】以微量点滴法将苯醚威施药于印鼠客蚤早3龄幼虫和未吸血新羽化成虫,采用组织学、显微摄影及统计学方法观察组织变化。【结果】经苯醚威作用后,印鼠客蚤的早 3 龄幼虫的表皮增厚、卵巢芽生殖细胞萎缩、睾丸芽精原细胞间质减少;未吸血新羽化成虫的睾丸塞消失快、唾液腺细胞破坏严重、中肠上皮细胞萎缩。【结论】(1)苯醚威通过干扰印鼠客蚤幼虫的变态,引起幼虫表皮、生殖芽异常改变,不能发育为成虫而死亡;(2)苯醚威可加速印鼠客蚤新羽化雄性成虫的睾丸塞吸收;(3)苯醚威可破坏印鼠客蚤新羽化成虫的唾液腺细胞,并引起中肠上皮细胞萎缩。     

不等单蚤和缓慢细蚤吸血前后脂肪和非特异性酯酶的组织化学研究

采用组织化学和显微摄影及定量图像分析技术,研究了不等单蚤Monopsyllus anisus (Rothschild)和缓慢细蚤Leptopsylla segnis (Schönherr) 在新羽化和吸血后发育过程中脂肪和非特异性酯酶的分布及活性。结果显示,两种新羽化蚤脂肪主要存在脂肪体中,非特异性酯酶则主要存在于唾液腺和神经链中。不等单蚤吸血后中肠脂滴增加,非特异性酯酶活性增强,随消化时间的延长脂滴逐渐减少,但不同消化时间非特异性酯酶活性差异不显著;而缓慢细蚤吸血后中肠未见或偶见少量脂滴,非特异性酯酶活性也有增强,但酶活性低于相同消化时间的不等单蚤。卵母细胞发育过程中脂滴增多,非特异性酯酶活性逐渐增强,且不等单蚤酶活性强于缓慢细蚤的。     

伯氏疟原虫PbPH表达特点和单克隆抗体传播阻断效应研究

制备抗PbPH单克隆抗体(mAb),检测伯氏疟原虫(Plasmodium berghei)PbPH的表达特点和传播阻断能力。主要通过蛋白免疫印迹(Western blot, WB)和间接免疫荧光法(Indirect immunofluoscent assay, IFA),确定PbPH的表达阶段以及抗PbPH单克隆抗体的特异性。1×10~6个P.berghei感染雌性BALB/c小鼠3 d后,尾静脉收集感染小鼠的血液,与鼠源的抗PbPH单克隆抗体/PBS对照,按照不同的稀释倍数(1?5、1?10、1?50)进行混合培养,观察15 min后,伯氏疟原虫配子体出丝中心数及24 h后动合子形成数的变化。WB和IFA检测抗PbPH单克隆抗体可以识别雌雄配子体、雄配子、雌配子/合子、retort和动合子的表面抗原。体外传播阻断实验中,与PBS对照组相比,在加入不同浓度(1?5、1?10)的抗PbPH单克隆抗体培养后,配子体出丝中心数分别减少了41.7%、32.7%,差异具有统计学意义(P0.05);动合子形成数分别减少了45.1%、14.8%,差异具有统计学意义(P0.05)。抗PbPH单克隆抗体可以有效阻断体外配子体出丝中心和动合子的形成,从而影响伯氏疟原虫在蚊体内的进一步发育和继续传播。     

丽斗蟋翅二型雄虫食物消化能力及消化酶活性比较

【目的】丽斗蟋Velarifictorus ornatus具明显的翅二型现象,为探讨翅型分化对丽斗蟋翅二型雄虫消化能力及中肠内消化酶活性产生的影响,对长翅型与短翅型雄虫食物消化能力及中肠内消化酶活性进行了检测比较。【方法】我们采取重量营养指数测定了羽化后12 d内丽斗蟋两型雄成虫增长量、相对增长率、取食量、食物利用率、近似消化率和食物转化率。为进一步明确丽斗蟋翅二型成虫食物消化能力与中肠内消化酶活性的关系,我们采用4种专用底物测定了中肠内用于分解蛋白质、脂肪和碳水化合物的总蛋白酶、胰蛋白酶、脂肪酶和淀粉酶的活性。【结果】结果表明,丽斗蟋两型雄虫取食量、食物转化率、食物利用率与增长量均无统计差异,但中肠内消化酶活性变化规律不同。成虫羽化后4 d时,长翅型雄虫中肠内总蛋白酶与胰蛋白酶活性显著高于短翅型雄虫,相反,羽化后0 d时,短翅型雄虫中肠内总蛋白酶与胰蛋白酶活性则显著高于长翅型雄虫,而羽化后12 d时,虽然短翅型雄虫总蛋白酶活性高于长翅型雄虫,但胰蛋白酶活性在两型雄虫间并无差异。成虫羽化后0 d时,两型雄虫脂肪酶活性无差异,但无论是羽化后4 或 12 d,长翅型雄虫中肠内脂肪酶活性皆显著大于短翅型雄虫。成虫羽化后4 d时,短翅型雄虫中肠内淀粉酶活性显著高于长翅型雄虫,而羽化后0与12 d时,两型雄虫间无显著差异。【结论】丽斗蟋翅二型雄虫食物消化能力无显著差异,但羽化后不同时间,中肠内消化酶活性存在差异,该差异可能与成虫羽化后不同时期,翅二型雄虫在飞行与繁殖投资中对不同能源物质的需求有关。     

Characterization and identification of exflagellation-inducing factor in the salivary gland of Anopheles stephensi (Diptera: Culicidae)

Gamete activation factor (GAF) induces exflagellation of Plasmodium microgametes. We found GAF in the salivary glands of female mosquitoes, Anopheles stephensi. The exflagellation was induced in a concentration-dependent manner in the supernatant of salivary gland's crude homogenate. The exflagellation-inducing activity in the salivary gland was higher than that in the midgut and the head. GAF in the salivary glands was found to be heat stable and low molecular weight (<3000 molecular weight). Analysis of the supernatant by capillary electrophoresis and UV absorbance profile showed that the salivary glands contained xanthurenic acid, which was previously identified as GAF in the head of A. stephensi. The exflagellation-inducing activity in the salivary gland declined immediately after a blood meal, implying that GAF was in the saliva, and was delivered into the midgut together with the blood and induced exflagellation in the midgut.     

The effects of blood feeding and exogenous supply of tryptophan on the quantities of xanthurenic acid in the salivary glands of Anopheles stephensi (Diptera: Culicidae)

Xanthurenic acid (XA), produced as a byproduct during the biosynthesis of insect eye pigment (ommochromes), is a strong inducer of Plasmodium gametogenesis at very low concentrations. In previous studies, it was shown that XA is present in Anopheles stephensi (Diptera: Culicidae) mosquito salivary glands and that during blood feeding the mosquitoes ingested their own saliva into the midgut. Considering these two facts together, it is therefore likely that XA is discharged with saliva during blood feeding and is swallowed into the midgut where it exerts its effect on Plasmodium gametocytes. However, the quantities of XA in the salivary glands and midgut are unknown. In this study, we used high performance liquid chromatography with electrochemical detection to detect and quantify XA in the salivary glands and midgut. Based on the results of this study, we found 0.28+/-0.05 ng of XA in the salivary glands of the mosquitoes, accounting for 10% of the total XA content in the mosquito whole body. The amounts of XA in the salivary glands reduced to 0.13+/-0.06 ng after mosquitoes ingested a blood meal. Approximately 0.05+/-0.01 ng of XA was detected in the midgut of nonblood fed An. stephensi mosquitoes. By adding synthetic tryptophan as a source of XA into larval rearing water (2 mM) or in sugar meals (10 mM), we evaluated whether XA levels in the mosquito (salivary glands, midgut, and whole body) were boosted and the subsequent effect on infectivity of Plasmodium berghei in the treated mosquito groups. A female specific increase in XA content was observed in the whole body and in the midgut of mosquito groups where tryptophan was added either in the larval water or sugar meals. However, XA in the salivary glands was not affected by tryptophan addition to larval water, and surprisingly it reduced when tryptophan was added to sugar meals. The P. berghei oocyst loads in the mosquito midguts were lower in mosquitoes fed tryptophan treated sugar meals than in mosquitoes reared on tryptophan treated larval water. Our results suggest that mosquito nutrition may have a significant impact on whole body and midgut XA levels in mosquitoes. We discuss the observed parasite infectivity results in relation to XA's relationship with malaria parasite development in mosquitoes.     

The role of the mosquito peritrophic membrane in bloodmeal digestion and infectivity of Plasmodium species.

Secretion and luminal formation of the peritrophic membrane (PM) were induced in female Anopheles stephensi and Aedes aegypti by feeding the mosquitoes on a warmed suspension of latex particles in Ringer's solution. The PM in A. stephensi was produced from apical secretion vesicles stored in the midgut epithelial cells and secreted into the lumen during feeding. In A. aegypti, the PM was formed de novo. When the latex feeding was followed 24 hr later by a meal of lyophilized pig blood, the 2 mosquito species exhibited very different modifications to their PM structure; in A. stephensi no PM was formed around the blood meal, whereas de novo synthesis of the PM in A. aegypti continued during the blood meal, with the resulting PM greatly thickened compared to the normal feeding. This artificial induction of PM formation was used as the basis to study the role of the PM in blood meal digestion and in infectivity of mosquitoes by the appropriate species of Plasmodium. The feeding of a latex suspension alone had no stimulatory effect on the 2 major midgut proteases, trypsin and aminopeptidase, in either species. After a blood meal alone, proteases rose to maximum activity at 30 hr and 24 hr after feeding in A. stephensi and A. aegypti, respectively. After double feeding, protease activities in both species were almost identical to those in blood-fed mosquitoes. Neither the absence of a PM (in A. stephensi) nor the presence of a thickened PM (in A. aegypti), therefore, has any effect on the ability of mosquitoes to digest a blood meal. Malaria infectivity, measured by oocyst counts, also was compared after normal and double feeding using infective blood meals. Infectivity of A. stephensi by Plasmodium berghei was unaffected by the presence or absence of the PM. The thickened PM produced by double feeding in A. aegypti caused a reduction of midgut infectivity by Plasmodium gallinaceum. These results suggest that the PM may act as a partial, but not an absolute, barrier to invasion of the midgut by the ookinete.     

Salivary gland proteins of the mosquito Culex quinquefasciatus

Several properties of the salivary glands of Culex quinquefasciatus mosquitoes were analysed. The amount of protein in female salivary glands increased from 0.26 microg on day one after emergence to about 1.4 microg on day seven. The major polypeptides found in the female salivary glands had molecular weights of 35.7, 28.3, and 20.5 kDa. Antibodies produced by mice immunized by bites of Culex quinquefasciatus female mosquitoes reacted with the 35.7 and 28.3 kDa polypeptides, showing that these molecules were secreted by mosquitoes during blood feeding. The salivary glands of C. Quinquefasciatus females displayed the same morphological and biochemical organization as that of Aedes aegypti mosquitoes, accumulating apyrase in the distal portions and alpha-glucosidase in the proximal portions of the gland. Arch.     

Pathology of Anopheles stephensi after infection with Plasmodium berghei berghei. I. Mortality rate.

The mortality of P. berghei-infected Anopheles stephensi females can be about 30% higher during the first three days than in normal blood-fed mosquitoes. As expected the mortality is higher after feeding on highly infected mice but also depends on the date of feeding and the temperature. Infected mosquitoes kept at 25 degrees C die more often than those kept at 21 degrees C. On the other hand sporozoite production needs the low temperature of 21 degrees C. So the sporozoite production rate falls with increasing temperature, and the mortality rate increases.     

Analysis of the sporogonic development of Plasmodium falciparum and Plasmodium berghei in anopheline mosquitoes

Basic knowledge of the sporogonic development of malarial parasites is crucial when evaluating the sporontocidal activity of antimalarial drugs or when determining why certain vectors are refractory to a particular parasite while others are competent vectors. We have developed a model which we have used to i) assess the sporogonic development of Plasmodium berghei ANKA in Anopheles stephensi and A. freeborni mosquitoes and ii) determine the effect of chloroquine on the sporogony of P. falciparum NF-54 in A. stephensi. Criteria used to assay sporogonic development include: i) number of oocysts present, ii) percentage of mosquitoes with oocysts, iii) time of release of sporozoites from the oocysts into the hemolymph, iv) time and degree of sporozoite invasion of salivary glands, and v) transmission (P. berghei) into vertebrate hosts. Parasite development in the mosquito is evaluated every other day, commencing on ca. day 7 post-feed (PF) and continuing until ca. day 22 PF. These detailed observations allow us to delineate the chronology of sporogonic development.     

Imaging movement of malaria parasites during transmission by Anopheles mosquitoes

Malaria is contracted when Plasmodium sporozoites are inoculated into the vertebrate host during the blood meal of a mosquito. In infected mosquitoes, sporozoites are present in large numbers in the secretory cavities of the salivary glands at the most distal site of the salivary system. However, how sporozoites move through the salivary system of the mosquito, both in resting and feeding mosquitoes, is unknown. Here, we observed fluorescent Plasmodium berghei sporozoites within live Anopheles stephensi mosquitoes and their salivary glands and ducts. We show that sporozoites move in the mosquito by gliding, a type of motility associated with their capacity to invade host cells. Unlike in vitro, sporozoite gliding inside salivary cavities and ducts is modulated in speed and motion pattern. Imaging of sporozoite discharge through the proboscis of salivating mosquitoes indicates that sporozoites need to locomote from cavities into ducts to be ejected and that their progression inside ducts favours their early ejection. These observations suggest that sporozoite gliding allows not only for cell invasion but also for parasite locomotion in host tissues, and that it may control parasite transmission.     

Egg development in the mosquito Anopheles albimanus

Summary

In the mosquito, Anopheles albimanus, previtellogenic egg development was completed by 48 h after emergence, and vitellogenic growth was completed by 36 h after a blood meal. Ecdysteroid levels reached a peak of 800 pg/female by 18 h, while vitellin levels rose to their maximum 36–48 h after a blood meal. Most of the ecdysteroids present in the female before 36 h behaved as ‘free’ hormone, while after 42 h the ecdysteroids were ‘conjugated’. Injection of 20-hydroxyecdysone into non-blood-fed females induced degeneration of the resting stage oocytes, but vitellogenin synthesis was detectable by autoradiography. Injection of 5 μ of 20-hy-hroxyecdysone into blood-fed decapitated females induced almost precisely normal levels of vitellin. Detailed analysis of the effect of decapitating blood-fed females suggested that the release of factors from the head (e.g., egg development neurosecretory hormone) occurs as an all-or-none phenomenon, and probably occurs twice.     

The effect of Plasmodium yoelii nigeriensis infection on the feeding persistence of Anopheles stephensi Liston throughout the sporogonic cycle.

Vector-borne parasites such as malaria have been shown to modify the feeding behaviour of their invertebrate hosts so as to increase the probability of transmission. However, evolutionary consideration of developmental changes in malaria within Anopheles mosquitoes suggests that the nature of altered feeding by mosquitoes should differ depending on the developmental stage of the parasite. We present laboratory evidence that the feeding persistence of female Anopheles stephensi towards a human host is decreased in the presence of Plasmodium yoelii nigeriensis oocysts (which cannot be transmitted), but increased when the malaria has developed into transmissible sporozoites in the salivary glands. In ten-minute trials, 33% of uninfected mosquitoes gave up their feeding attempt before the test period had ended, 53% of those harbouring oocysts had given up, but only 20% of those infected with sporozoites gave up by this time. We conclude that changes in feeding behaviour of mosquitoes mediated by parasite infection are sensitive to the developmental stage of the parasite and that these changes have important implications for malaria epidemiology.     

The sporogonic cycle of Plasmodium reichenowi

Plasmodium reichenowi, a malarial parasite of the chimpanzee, was infective to Anopheles freeborni, Anopheles quadrimaculatus, Anopheles stephensi, Anopheles maculatus, Anopheles dirus, and Anopheles culicifacies mosquitoes. Anopheles gambiae and Anopheles albimanus were not infected. Mean oocyst diameters of P. reichenowi were smaller than those of the other chimpanzee parasite, Plasmodium schwetzi. Sporozoites were present in the salivary glands of An. freeborni at 15 days when held at 25 to 26 C.