Physiological basis of host susceptibility of Florida mosquitoes to Dirofilaria immitis.

Young females of seven species of Florida mosquitoes were fed a meal of dog blood infected with Dirofilaria immitis to repletion to study the physiological mechanisms which control susceptibility and resistance in these mosquitoes. Various species of mosquitoes showed different grades of susceptibility. In all mosquitoes, microfilariae reached the midgut immediately after ingestion. Their movement from midgut to the specific host tissue—the Malpighian tubules—was either facilitated or inhibited depending on the presence or absence of anticoagulins in the salivary glands of these mosquitoes. In Anopheles quadrimaculatus, Aedes taeniorhynchus, and Aedes sollicitans, microfilariae move freely from the midgut to the Malpighian tubules, because of the presence of substantial amounts of anticoagulins in their salivary glands, and 30 to 60 mf/female developed normally to an infective stage. Very few microfilariae reached the tubules of Mansonia titillans as most of them were defaecated within a very short time after ingestion. In Aedes aegypti, Culex nigripalpus, and Culex pipiens quinquefasciatus movement of microfilariae from the midgut to the Malpighian tubules was obstructed by the coagulation of blood soon after ingestion. Coagulation of blood was followed by formation of oxyhaemoglobin crystals in C. nigripalpus and C. p. quinquefasciatus. It is suggested that secretions from symbiotic bacteria in the midgut of these mosquitoes lyse ingested red blood cells, and the released haemoglobin is oxidized to oxyhaemoglobin crystals which hinder the further movement of microfilariae and kill them.Microfilariae developed normally in A. quadrimaculatus, thus making them potentially the most susceptible mosquitoes, even though these mosquitoes did not survive to be effective potential vectors. A few microfilariae or their later developmental stages were melanized in the tubules of most A. sollicitans and A. taeniorhynchus, but the numbers of melanized stages were too few to affect the vectoring potentials of these species. In 20 per cent of A. sollicitans, 60 per cent of M. titillans, and ca. 80 per cent A. aegypti substantial numbers of the microfilariae after reaching the Malpighian tubules did not advance beyond the prelarval stage, and very few microfilariae developed successfully in the remaining mosquitoes. Very few microfilariae reached the Malpighian tubules of a small percentage of C. nigripalpus and C. p. quinquefasciatus and developed normally. The vectoring potentials of A. sollicitans, M. titillans, A. aegypti, and both Culex species were greatly hampered. These studies suggested that host-specificity of mosquitoes to D. immitis infection is controlled by the presence or absence of secondary physiological factors in their digestive tracts or in the Malpighian tubules.     

Size‐dependent insect flight energetics at different sugar supplies

Under most circumstances, large body size confers a higher fitness and is positively selected, whereas selection against large size is empirically poorly documented. Physiologically, according to the ¾ power law, larger animals have lower relative but higher absolute energy demands, such that large body size may become disadvantageous, particularly under fast locomotion in food‐limited environments. After a period of initial feeding on different sugar concentrations, we investigated size‐dependent energy content (reserves) at baseline and of females unflown (i.e. resting) or flown for 18 h in two (replicate) insect species: the yellow dung fly Scathophaga stercoraria and the yellow fever mosquito Aedes aegypti. Tethered adults of various sizes were tested in a flight mill. In both species, teneral glycogen, sugar, and lipid content increased with sugar availability, and isometrically or even hyper‐allometrically (slope > 1) with body size. Activity treatment also revealed the expected consumption effects. Both species increased their flight distance with sugar supply, although only larger mosquitoes flew longer. Crucially, larger females of both species disproportionately exhausted more glycogen and sugars (but not lipid) during flight. The mosquitoes appeared to adjust their flight more finely to their size‐dependent energy reserves at all sugar availabilities, whereas, in the dung flies, size‐dependent energy demands were detectable only with a low but not with an overly high sugar supply. Although we found a greater absolute and relative locomotory energy demand for the larger flies, which is in agreement with interspecific patterns in insects, this was (more than) compensated by their greater baseline energy reserves, resulting in the greater net flying capacity of larger individuals. Consequently, we found no evidence for energetic mechanisms limiting the performance of large flying insects under food limitation. The differences between the two species presumably relate to mosquitoes inherently being long distance flyers and dung flies being short distance flyers. © 2012 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, ●● , ●●–●●.     

Long-term regulation of sucrose intake by the female mosquito, Aedes taeniorhynchius

Newly emerged female Aedes taeniorhynchus were maintained in cups provided with potometers containing 5, 10, and 25% sucrose solutions. Mortality and the ab lib. intake of sucrose were recorded daily and groups were sampled at regular intervals for analysis of energy reserves.After a week of relatively substantial intake of sucrose at all three concentrations, a constancy in daily intake at much lower levels was observed during the next 4 weeks. The caloric intake was highest in the 25% sucrose solution group. There were no differences in mortality rates at these concentrations. Glycogen (0·7–1·0 cal/female) and triglycerides (3·5dash4 cal/female) reserves reached maximum levels after the first week and stabilized thereafter on all three concentrations. In this species the stabilization in intake of sucrose solution after the first week is correlated with the maximum accumulation of stored energy reserves as opposed to decreased activity or subtle changes associated with senescence.     

Metabolism of Glycogen and Neutral Lipids by Aphelenchus avenae and Caenorhabditis sp. in Aerobic,Microaerobic and Anaerobic Environments

Starving Aphelenchus avenae survived 3-4 weeks in microaerobic and anaerobic environments, but Caenorhabditis sp. survived less than 80 hr. Aerobically, both nematodes metabolize neutral lipid reserves: there was no microaerobic ( <5% O₂) or anaerobic neutral lipid catabolism. Early in anaerobiosis both nematodes utilized endogenous glycogen. Caenorhabditis sp. depleted the glycogen and died. A. avenae under oxygen stress longer than 120 hr entered cryptobiosis, during which there was neither measurable O₂ uptake nor glycogen or neutral lipid utilization, Only when re-aerated, did A. avenae recover and resume "''normal" metabolism.     

Effect of fasting on glycogen metabolism and activities of glycolytic and gluconeogenic enzymes in the mudskipper Boleophthalmus boddaerti

During starvation, muscle glycogen in Boleophthalmus boddaerti was utilized preferentially over liver glycogen. In the first 10 days of fasting, the ratio of the active‘a’form of glycogen phosphorylase to total phosphorylase present in the liver was small. During this period, the active‘I’form of glycogen synthetase increased in the same tissue. In the muscle, the phosphorylase‘a’activity declined during the first 7 days and increased thereafter while the total glycogen synthetase activity showed a drastic decline during the first 13 days of fasting. The glycogen level in the liver and muscle of mudskippers starved for 21 days increased after refeeding. After 6 and 12 h refeeding, liver glycogen level was 8·5 ± 2·3 and 6·9 ± 4·5 mg·g wet wt ¹, respectively, as compared to 5·8 ± l·6mg·g wet wt ¹ in unfed fish. Muscle glycogen level after 6 and 12 h refeeding was 0·96±0·76 and 0·82 ± 0·50 mg·g wet wt ¹, respectively, as opposed to 0·21 ± 0·12 mg·g wet wt ¹ in the 21-days fasted fish. At the same time, activities of glycogen phosphorylase in the muscle and liver increased while the active‘I’form of glycogen synthetase showed higher activity in the liver. Since glycogen was resynthesized upon refeeding, this eliminated the possibility that glycogen depletion during starvation was due to stress or physical exhaustion after handling by the investigator. Throughout the experimental starvation period, the body weight of the mudskipper decreased, with a maximum of 12% weight loss after 21 days. Liver lipid reserves were utilized at the onset of fasting but were thereafter resynthesized. Muscle proteins were also metabolized as the fish were visibly thinner. However, no apparent change in protein content expressed as per gram wet weight was detected as the tissue hydration state was maintained constant. The increased degradation of liver and muscle reserves was coupled to an increase in the activities of key gluconeogenic enzymes in the liver (G6Pase, FDPase, PEPCK, MDH and PC). The increase in glucose synthesis was possibly necessary to counteract hypoglycemia brought about by starvation in B. boddaerti.     

Water exchange and effect of water vapour activity on metabolic rate in the dust mite, Dermatophagoides.

Oxygen consumption by Dermatophagoides farinae was found to vary with temperature and water vapour activity. The relationship between temperature and O₂ consumed/hr per mite was first order. Q₁₀'s were equal to exp (k 10°C) and were found to be 3·04 and 2·49 for 1 to 6 and 6 to 22 hr monitoring periods respectively. A significant difference between O₂ consumed/hr per mite for 1 to 6 and 6 to 22 hr monitoring periods was found. Inactivity of mites explained 29·6 and 31·8 per cent of this reduction. Dehydrating conditions and reduced permeability of the water and gas exchange surface explained a further reduction to 58·7 and 60·8 per cent.     

Effects of sucrose on blood avidity in mosquitoes.

Within 10 min after engorging on 10% sucrose, most females of Aedes aegypti do not seek a human blood meal, but remain quiescent and unresponsive to a human hand in a 1 ft³ cage. The duration of this inhibition in blood avidity varies greatly among individuals, but may last for 2 to 5 hr after drinking sucrose. There was no specific correlation between abdominal distension and blood avidity. When females engorged on varying concentrations of sucrose, it was found that as the concentration increased, fewer mosquitoes would take blood when it was offered after 1 hr. Only 25% of the females which engorged on 0·5 to 1 M sucrose took human blood at this time. As the concentration of sucrose increases, there is a marked decrease in spontaneous flight activity during the first hour after feeding. When unfed females are injected with 10% sucrose or trehalose, none of them took blood for 3 hr, whereas 50% of the saline-injected controls fed on blood.     

Culicidae (Diptera) selection of humans,chickens and rabbits in three different environments in the province of Chaco, Argentina

Studies were conducted to determine the selection of humans, chickens and rabbits by Culicidae in three different environments in the province of Chaco, Argentina. Mosquitoes were collected fortnightly using cylindrical metal traps containing animal bait (chickens and rabbits). The mosquitoes were collected between June 2001-May 2002. During the same period and with the same frequency, mosquitoes biting the human operators of the traps were collected during the first 15 min of exposure within different time intervals: from 09:00 am-11:00 am, 01:00 pm-03:00 pm, 05:00 pm-07:00 pm and 09:00 pm-10:00 pm. A total of 19,430 mosquitoes of 49 species belonging to 10 genera were collected. Culex species mainly selected chicken bait and Wyeomyia species selected rabbit bait. Ochlerotatus and Psorophora species were more abundant in rabbit-baited traps. Anopheles triannulatus, Coquillettidia nigricans, Ochlerotatus scapularis, Mansonia titillans and Psorophora albigenu showed a strong attraction for human bait. The Anopheles, Coquillettidia, Culex and Mansonia species were more active between 05:00 pm-09:00 pm, while Ochlerotatus, Psorophora, Haemagogus and Wyeomyia were most active from 09:00 am-07:00 pm. This study provides additional information about the biology and ecology of arbovirus vectors in Chaco.     

Activation of vitellogenin synthesis in the mosquito Aedes aegypti by ecdysone

Injected β-ecdysone was found to induce the synthesis of yolk protein (vitellogenin) in adult female Aedes aegypti without a blood meal. After injection of 5 μg ecdysone per mosquito, vitellogenin constituted 80 per cent of the total protein secreted by explanted fat body, a proportion comparable to that produced by fat body from blood-fed females. Moreover, the time course of induction of vitellogenin synthesis in ecdysone-injected mosquitoes was similar to that triggered by a blood meal. Response to ecdysone is dosedependent: 0·5 μg per female was required to stimulate synthesis to 50 per cent of the level found 18 hr after a blood meal. Ecdysone was effective in decapitated or ovariectomized mosquitoes, and also when applied directly to fat body preparations in vitro. Thus it appears that ecdysone acts directly on the fat body to induce specific protein synthesis, as does the vitellogenin stimulating hormone (VSH) from the ovary of blood-fed mosquitoes. These results suggest that ecdysone can replace VSH in inducing vitellogenin synthesis in the unfed mosquito.     

Regulation of carbohydrate metabolism and flight performance by a hypertrehalosaemic hormone in the mosquito Anopheles gambiae

The role of adipokinetic hormones (AKHs) in the regulation of carbohydrate and lipid metabolism and flight performance was evaluated for females of the African malaria mosquito, Anopheles gambiae. Injection of various dosages of synthetic Anoga-AKH-I increased carbohydrate levels in the haemolymph and reduced glycogen reserves in sugar-fed females but did not affect lipid levels. Anoga-AKH-I enhanced the flight performance of both intact and decapitated sugar-fed females, during a 4 h flight period. Anoga-AKH-II had no effect on carbohydrate or lipid levels or flight performance, thus its function remains unknown. Targeted RNA-interference lowered Anoga-AKH receptor expression in sugar-fed females, consequently injections of Anoga-AKH-I failed to mobilize glycogen reserves. Taken together, these results show that a primary role for the neurohormone, Anoga-AKH-I, is to elevate trehalose levels in the haemolymph of female mosquitoes.     

Flight performance of the malaria vectors Anopheles gambiae and Anopheles atroparvus.

The flight potential and metabolism of two malaria vectors, Anopheles gambiae s.str. and An. atroparvus, were analyzed on flightmills. The flight distance, the flight time, and individual flight activities of females were recorded during 22 h flight trials. The glycogen and lipid before flight, after flight, and of unflown controls were measured for starved, sugar-, or blood-fed females. Maximal flight distances of An. gambiae were 9 km when sugar-fed and 10 km when blood-fed, while in starved females it was below 3 km and the average speed was around 1 km/h. In Anopheles atroparvus, the maximal flight distances were 10-12 km when sugar-fed, 4.5 km when blood-fed, and below 3.5 km when starved, with an average speed of 1.3 km/h. Flight performances consisted of 1-4 h intervals of continuous flights, but mainly of bouts shorter than one h, randomly distributed during the long flight trials in both species. An. gambiae utilized an average of 47% of its pre-flight carbohydrate reserves for survival and 38% for flight at a rate of 0.07 cal/h/female. After a blood meal they utilized 11% for survival and 61% for flight at a rate of 0.04 cal/h. At the same time, 25% of the pre-flight lipid was mobilized for flight at a rate of 0.09 cal/h when sugar-fed and 22% when blood-fed at a rate of 0.06 cal/h; lipid was barely mobilized for survival. An. atroparvus differed: carbohydrate mobilization was 28% for survival and 41% for flight at a rate of 0.15 cal/h when sugar-fed; lipid mobilization for flight was only 13% at a rate of 0.06 cal/h. After a blood meal only 2% of the pre-flight lipid was used (0.02 cal/h). The contribution of carbohydrate reserves for flight metabolism at the high rate of 0.21 cal/h could not be fully elucidated because its decrease coincided with a pronounced resynthesis from the blood meal. An. atroparvus always depended on sugar meals for its flight activities and barely utilized lipid reserves. An. gambiae was independent of sugar sources for strong flights due to its early blood feeding and because of its equicaloric lipid mobilization during flights. Strong evidence for lipid oxidation during its flight is discussed.     

Boreal forest dynamics in north-eastern Sweden during the last 10,000 years based on pollen analysis

A pollen record obtained from a 2.2-m sediment succession deposited in a small lake in the province of Västerbotten, north-eastern Sweden, reveals the presence of continuous forest cover since 8,500 calendar years before present (cal b.p.). Forest with abundant Pinus (pine) and Betula (birch) initially colonized the area, followed by a dominance of deciduous trees, primarily Betula, from ca. 8,000 to ca. 3,200 cal b.p. Pollen accumulation rates of Quercus (oak), Ulmus (elm) and Tilia (linden) suggest the possible local presence of these thermophilous tree species during this period. The climate gradually became colder and moister around 3,500 cal b.p. and an increased abundance of Sphagnum spores indicates paludification. Picea (spruce) became established around 3,200 cal b.p. and less than 500 years later this was the dominant tree species around the lake. The fire frequency as inferred from charcoal particles exhibits a general increase from ca. 3,000 cal b.p. with subsequent charcoal accumulation maxima at around 2,800 cal b.p., 1,700 cal b.p. and in recent time. The human influence on vegetation was significant during the last 200–300 years. Soil erosion increased substantially and fern spores amount to ca. 55% of the total pollen assemblage in the uppermost samples. These results suggest an extensive anthropogenic impact on the local forest ecosystem, with abundant logging, burning and ditching in the vicinity of the lake. Independent evidence of sub-recent human-induced environmental change is provided by historical accounts. Complementary information on catchment soil development and aquatic nutrient status was provided by records of magnetic susceptibility and elemental carbon, and nitrogen contents obtained from the same sediment core.     

Brugia pahangi: effects upon the flight capability of Aedes aegypti.

Aedes aegypti mosquitoes were adversely affected by infections of the filarial worm Brugia pahangi. Infected mosquitoes flew significantly shorter distances and showed marked reductions in total flight time during 24-hr flight mill tests compared to uninfected controls. Total flight range and duration flown by infected mosquitoes remained relatively constant throughout the infection process, while control mosquitoes flew further and longer with increasing time after their blood meal. Furthermore, a significantly greater number of infected mosquitoes either died or were rendered incapable of flight. Of flying and nonflying mosquitoes with 6-day-old or older infections dissected for parasite burdens, the nonflying group contained significantly more worms. Results of this study indicate that developing filarial larvae within this mosquito vector reduce its ability to survive and to transmit its infection by reducing its flight capabilities. Conclusions from this study relate only to A. aegypti homozygous for the gene fm which is fully susceptible to this filarial parasite.     

Dependence of the life span of the honeybee (Apis mellifica) upon flight performance and energy consumption

Summary The life span of worker-honeybees is determined by the duration of the hive-period and of the foraging period (Figs. 1,2). The duration of the forgaing period is regulated in the following way: Total flight performance of the individual bee seems to be fixed. Daily flight performance strongly affects total flight duration. High daily flight performance decreases maximal flight duration and vice versa.Foragers accumulate the highest glycogen reserves in the flight muscles compared to other stages (Figs. 3, 4). They use these reserves to overcome starvation or when growing old. Young foragers are able to restore glycogen reserves after sugar intake, whereas old foragers were found to have a reduced glycogen synthesizing ability (Fig. 5).The results indicate that bees exhaust their energysupplying mechanisms after a definite total flight performance.     

18,000 years of grassland evolution in the summer rainfall region of South Africa: evidence from Mahwaqa Mountain,KwaZulu-Natal

A palynological and sedimentological record from the Mahwaqa Mountain in KwaZulu-Natal, South Africa, provides evidence of the vegetation dynamics in this part of the Grassland Biome during the last c. 18,000 years. The wetland is located at 1,850 m on an isolated outlier of the Ukhahlamba–Drakensberg Mountain range on an ecotone along a climatic gradient. The vegetation responded to humidity and temperature changes during the late Pleistocene and Holocene. The period c. 18,000–13,500 cal. bp is characterized by high Ericaceae and Restionaceae percentages and decreasing values of charred particles, indicating cool conditions. Around 13,500–8,500 cal. bp, Ericaceae were gradually replaced by Poaceae, signaling climate warming. Growing environmental wetness during the same time period is inferred from Phragmites-type and Cliffortia pollen percentages. Since c. 8,500 cal. bp, Cliffortia, Restionaceae, and Phragmites-type percentages have maintained low levels. Ericaceae were almost completely replaced by grasses and Asteraceae by c. 7,500 cal. bp. All indications are that warm and fluctuating moisture conditions followed until 4,600 cal. bp but they became driest between c. 4,600 and 3,500 cal. bp, when high Asteraceae, Pentzia-type and Scabiosa percentages were prominent. From c. 3,500–800 cal. bp, the increase of sedges, Aponogeton and grass pollen (including Phragmites-type) at the expense of Asteraceae pollen suggests the return of slightly more humid conditions. Since c. 1,000 cal. bp an increase of water demanding Podocarpus and Cliffortia occurred. Pine pollen indicates the recent introduction of alien plants in the 19th and 20th centuries.     

Flight energetics in relation to sexual differences in the mating behaviour of a mayfly,Siphlonurus aestivalis

Summary Mayflies (Ephemeroptera) are known to have short adult life-spans. Adults are unable to feed, and they utilize reserves stored during their aquatic larval stage. Energy reserves (fat, glycogen, and free sugars) of mature larvae, subimagoes and imagoes of both sexes of Siphlonurus aestivalis Eaton were compared. All the stages of both sexes had low glycogen and free sugar contents, and the only significant change occurred during the transformation of the mature larva to subimago when almost all the reserves of free sugars were used up. Glycogen and free sugars may serve as energy sources permitting individuals to swim and fly out of the water during emergence. Fat made up most of the energy reserves of mature larvae and was the main source of energy used during the final development of both sexes. Young adult males had high fat reserves which they used as a source of energy for their swarming flights. In contrast, females did not seem to use a significant amount of fat for flight. This difference is probably related to the different mating strategies of the sexes in this species. Males perform long flights waiting for females, whereas females perform only brief flights to mate and reproduce.     

Histopathology of a nuclear polyhedrosis infection in Aedes epactius with observations in four additional mosquito species

Aedes epactius larvae were utilized to study the infection sequence of the nuclear polyhedrosis virus (NPV) from Aedes sollicitans. From 30 min to 6 hr postinoculation, polyhedra and many free virions were observed in the larval midgut lumen. Penetration of the midgut cells by virions was not observed. The first infected nuclei were observed 12 hr postinoculation. Nucleocapsids initially exhibited electron translucent cores which became electron dense before the nucleocapsids acquired an envelope. Envelope acquisition occurred through a process of de novo membrane morphogenesis. Occlusion of the singly embedded virions began by 18 hr postinoculation with the mature rough-surfaced polyhedra averaging approximately 1 by 2 μm. Unusually long nucleocapsids (approximately two or three times the length of other nucleocapsids) were only observed in late infection period nuclei. There was no evidence that long nucleocapsids represented an early developmental stage for nucleocapsids of standard length. Infection was restricted to midgut nuclei and gastric caecae cells. Infected early instar A. epactius larvae became moribund 36 to 40 hr postinoculation and infected midgut nuclei were observed to undergo lysis. The late stages of NPV infection were observed in larvae of A. annandalei, Wyeomyia smithii, Toxorhynchites brevipalpus, and Eretmapodites quinquevittatus. Virion development and occlusion in these species was basically identical to the sequence observed in A. epactius larvae.     

The fine structure of peritrophic membranes of the blowfly, Calliphora erythrocephala, grown in vitro under different conditions.

The formation of peritrophic membranes (PM) in vitro was studied in a flow chamber in order to avoid the accumulation of metabolic substances during prolonged incubation. During the first 8 to 10 hr of incubation the production of PM was nearly constant—3·5 ± 1·4 mm PM/hr. After about 10 hr it decreased and stopped after about 35 hr. Between 1 and 8 hr after the beginning of incubation the width of the periodic crossband pattern reached or nearly reached the values found in PM which had formed in vivo; afterwards it decreased more and more. During the first 20 min of incubation a ‘disturbed zone’ of PM without any regular crossband pattern is formed.In the cardia of adult Calliphora erythrocephala there are three formation zones forming three PM of different fine structures. The fine structure of PM 1 to 3 formed in vitro during the first 6 to 8 hr of incubation in Leloup's medium 1, with an osmolarity of 340 mOsmol, a pH of 6·8, and a temperature of 27°C, does not differ from the PM grown in vivo. PM 1–3 grown in vitro in Tyrode's solution with added glutamine or in Leloup's medium with added β-ecdysone show a considerable increase in thickness and a disturbed formation of the electron dense layer of PM 1.     

Holocene mangrove dynamics and environmental change in the Rufiji Delta, Tanzania

Holocene mangrove dynamics are reconstructed from pollen, sediment and radiocarbon analyses of three cores (ANR, BNR, CNR) located across a 20 km transect in the Rufiji Delta, Tanzania. At the base of the sediment sequence, dated to about 5600 cal. year b.p., the mangroves which are present suggest a low intertidal ecosystem in response to wet conditions and a higher sea level than at the present day. After around 5600 cal. year b.p. in core BNR, mangroves retreated seaward probably due to a lower sea level and drier environmental conditions. At around 4640 cal. year b.p., mangroves shifted landward suggesting a phase of sea level rise. In the late Holocene, mangroves became established at higher elevations of the Rufiji Delta, which is now a paddy field. Mangrove taxa decreased after 1170 cal. year b.p., suggesting drier conditions and less inundation frequency, possibly due to a lower sea level. Marked vegetation changes from mangroves to terrestrial vegetation occurred after around 750 cal. year b.p., possibly related to sea level regression and/or a desiccation phase recorded during the late Holocene. Paddy fields replaced mangroves in the landward part of the transect, reflecting an increase in human settlement in this area, a trend that continues to the present day. The recent decrease of mangrove species, particularly Rhizophora mucronata, could suggest less inundation by saline water and a lower sea level, although these changes may also be due to human activities during the last millennia as indicated by charcoal analysis.     

Diapause hormone action in silkworm ovaries incubated in vitro; 14C-trehalose incorporation into glycogen

Developing ovaries from pharate adults of the silkworm, Bombyx mori, were incubated in a medium containing ¹⁴C-trehalose or ¹⁴C-glucose, and the effects of diapause hormone on the incorporation of these isotopes into ovary glycogen were studied. The rates of incorporation of ¹⁴C-trehalose remained unchanged in ovaries incubated for 36 hr when the medium was renewed at intervals of 12 hr, and showed saturation kinetics against the concentration of the sugar in the medium, giving apparent K_m values of 6·0 mM for trehalose.There was no difference in ¹⁴C-glucose incorporation by ovaries grown in the presence (+SG) and absence (?SG) of the suboesophageal ganglion (SG). However, when ¹⁴C-trehalose was used as a substrate for glycogen synthesis, there was a marked difference in the incorporation between them, i.e. the incorporation was more than 60 per cent higher in +SG ovaries than that in ?SG ovaries. Increased incorporation of ¹⁴C-trehalose was also observed in ovaries from SG-removed pharate adults which received an injection of diapause hormone preparations. Maximum stimulation rates (about twofold) appeared 36 hr after the injection. Further, comparable effects on ¹⁴C-trehalose incorporation were observed in ovaries which were incubated with diapause hormone preparations added in vitro.These data are discussed in relation to the hormonal regulation of trehalase activity in ovaries.