Hemolymph pH of the larvae of three species of mosquitoes,and the effect of Romanomermis culicivorax parasitism on the blood pH of Culex pipiens

The hemolymph pHs of late fourth-instar Culex pipiens, Aedes taeniorhynchus, and Toxorhynchites amboinensis were measured with pH-sensitive glass microelectrodes and were slightly alkaline; pH 7.51, 7.62, and 7.37, respectively. The hemolymph pH remained relatively constant during the development of C. pipiens larvae through the third and early and late fourth instars. The hemolymph pH in C. pipiens larvae parasitized with the mermithid nematode Romanomermis culicivorax was unaltered. These measurements provide an approximate pH (ca 7.4) which is normal for the hemolymph of larval mosquitoes, and should be useful for further development of a culture medium for R. culicivorax.     

Romanomermis culicivorax parasitism and the development, growth, and feeding rates of two mosquito species

The dry weight, development, and feeding rates of Culex pipiens and Toxorhynchites amboinensis larvae infected with the nematode, Romanomermis culicivorax, were measured and gross conversion efficiences were calculated. The weight of C. pipiens larvae infected at two different inoculum levels did not differ significantly from controls until day 6 postinfection (PI). Infected larvae of T. amboinensis were significantly lighter than controls at days 2, 4, and 6 PI. The rate of larval mosquito development was slowed after day 3 in parasitized individuals of both mosquito species. Infection significantly retarded the interval feeding rate of the filter-feeding C. pipiens throughout development. Infected T. amboinensis larvae consumed significantly fewer prey larvae of C. pipiens than controls. Calculation of gross conversion efficiency (GCE) showed that lightly infected C. pipiens larvae had an elevated GCE early in the infection but were less efficient relative to controls after 4 days PI. Lightly parasitized T. amboinensis had a lower total GCE than controls.     

Changes in body tissues and hemolymph composition of Culex pipiens in response to infection by Romanomermis culicivorax

Hemolymph composition of fourth instar larvae of an autogenous strain of Culex pipiens was examined to determine the effects of parasitism by a mermithid nematode, Romanomermis culicivorax. Mosquitoes were reared under two different pH regimens: 4.5 and 7.3. Wet and dry weight of infected mosquitoes reared at either pH were significantly lower than controls. The effects of parasitism in the development of C. pipiens were evaluated from paraffin sections of mosquito larvae 2, 4, and 6 days postinfection. At 2 days postinfection, the infected larvae showed no apparent effects of parasitism; at day 4, the fat body tissue was reduced and imaginal disc development was retarded; and at day 6, parasitized mosquitoes were smaller in cross section, fat body tissue was found only in isolated clumps, and there was a complete absence of imaginal discs. Concentrations of total carbohydrates in hemolymph from infected fourth instar mosquitoes reared at pH 7.3 were reduced. Trehalose and glucose were each reduced by more than half. Total α-amino nitrogen was significantly lower in infected mosquitoes reared at pH 7.3. However, total amino acid concentrations for hemolymph from control and infected larvae reared at pH 7.3 were the same. Methionine sulfoxide decreased 63% and proline increased 2.5 times in infected mosquitoes. Hemolymph protein concentrations were reduced 80% in infected mosquitoes reared at both pHs. The number of hemolymph proteins also declined from 35 to 22 during infection. Two host proteins, 82,000 and 158,000 daltons, remained prominent throughout the mermithid infection.     

The effect of nematode parasitism on the osmolality and major cation concentration in the haemolymph of three larval mosquito species

Parasitism by a mermithid nematode, Romanomermis culicivorax, causes severe depletion of haemolymph carbohydrates and proteins in mosquito larvae. We undertook a study to determine if haemolymph osmolality and cation concentrations were affected also by mermithid parasitism. The haemolymph osmolality of R. culicivorax-infected and control Aedes taeniorhynchus and Culex pipiens fourth-instar larvae was not significantly different. However, the haemolymph osmolality decreased significantly in infected Anopheles quadrimaculatus. Each mosquito species demonstrated significant alterations in the haemolymph concentration of at least one cation when infected although the cation concentrations affected differed for each species. The changes observed were statistically significant but the magnitude of change was not great. Overall, despite the severe nutritional burden of the mermithid nematode, these species of mosquito larvae can continue to maintain osmoregulation.     

中华甲虫蒲螨寄生双条杉天牛幼虫血淋巴变化

为了解中华甲虫蒲螨Pyemotes zhonghuajia Yu and Zhang对双条杉天牛Semanotus bifasciatus(Motschulsky)的致死过程,将已发育成熟的中华甲虫蒲螨膨腹体移入放有双条杉天牛2龄幼虫的玻璃指形管中,在蒲螨的一个生活史60h内,观察其取食行为,并在移入后每隔12h用考马斯亮蓝法分别测定天牛幼虫和蒲螨血淋巴中总蛋白质含量。结果显示,蒲螨蛋白含量总体呈现上升的趋势,在36～48h上升幅度最大,达61.78%;而天牛幼虫血淋巴总蛋白质含量则呈总体下降趋势,与对照相比,在36～48h之间下降最为明显;蒲螨总蛋白质含量在24h后处理组与对照组差异均为极显著,天牛幼虫总蛋白质含量在48h后处理组与对照组差异显著,这表明蒲螨在24h已搜寻到寄主开始取食,且取食效果明显,而天牛幼虫在48h后表现为麻痹致死,36～48h蒲螨取食对天牛致死起关键作用。12h时处理组的双条杉天牛幼虫总蛋白质含量比对照组略高,推测是由于幼虫对外寄生物蒲螨具有一定的免疫防御反应。     

Transcervical endometrial biopsy in the baboon: Effects on the menstrual cycle and relations to protein and dry matter content and histology

The baboon has been used increasingly for reproductive studies. While hormonal regulation of the menstrual cycle and ovulation as well as the endocrinology of gestation have been reported, little information is available describing endometrial parameters. It is the purpose of this paper to describe the ease with which repeated transcervical biopsies can be performed, to describe baseline endometrial protein and dry weight data and to demonstrate that the biopsy procedure itself does not significantly affect the baboons' ability to continue normal menstrual cycle function. Endometrial biopsy samples were taken throughout the menstrual cycle under light ketamine anesthesia. Protein and dry weight contents were determined. Endometrial biopsies Protein and dry weight contents were determined. Endometrial biopsies averaged 25 mg (wet weight) and contained 7.54% protein and 16.3% dry matter. The formulas (Y = a + bx) which expressed the linear relationships between wet weight (mg), protein (μg) and dry matter (μg) content and the correlation coefficients (r) were as follows: between wet weight and protein content – wet weight = 5.58 + 10.0 (protein), Sxy = 4.83, r = 0.883; between wet weight and dry weight – wet weight = 1.99 + 7.94 (dry weight), Sxy = 4.52, r = 0.904; between protein and dry weight – protein = 0.446 + 0.446 (dry weight), Sxy = 4.82, r = 0.870. All three linear regression coefficients were statistically significant (P < 0.001). No significant cyclical patterns in either protein or dry matter content were demonstrable throughout the menstrual cycles. The average length of all nonbiopsy cycles was 32.4 ± 2.7 days and 32.8 ± 3.6 days for those in which biopsies were taken. Similarly, follicular and luteal phase lengths for nonbiopsy and biopsy cycles were 15.4 ± 2.3 and 15.5 ± 2.8 days and 16.9 ± 2.2 and 17.2 ± 3.2 days, respectively. The time required for sex-skin swelling to decrease from maximum to minimum during the luteal phase was shorter, but the quiescent stage was equally lengthened. It was concluded that the endometrium of the baboon was easily accessible for study without causing serious alterations in menstrual cycle function. These studies further demonstrate the potential of the baboon as a model o reproductive studies. In fact, the baboon may well the only practical primate model available for endometrial studies.     

Susceptibility of ten species of mosquito larvae to the parasitic nematode Romanomermis iyengari and its development

Ten species of mosquitoes (Diptera: Culicidae) from five genera were exposed to preparasites of the tropical mermithid nematode species Romanomermis iyengari (Welch) (Nematoda: Mermithidae), a strain isolated in 1978 from Pondicherry. By exposing mosquito larvae during the second instar, nematode infection was invariably lethal, the rate being highest in Culex sitiens Wiedemann (95%) followed by Cx. quinquefasciatus Say (90%), Aedes aegypti (L.) (79%), Anopheles subpictus Grassi (64%), Ae. albopictus (Skuse) and Armigeres subalbatus Coquillett (62%), Cx. tritaeniorhynchus Giles (57%), Mansonia annulifera (Theobald) (46%), An. stephensi Liston (40%) and An. culicifacies Giles (36%). When fourth-instar larvae were exposed, the infection was highest in Ar. subalbatus (66%), followed by An. stephensi (52%), Cx. quinquefasciatus (47%), Ae. aegypti and An. subpictus (42%), Ae. albopictus (30%), An. culicifacies (29%), Cx. sitiens (24%), Cx. tritaeniorhynchus (19%) and Ma. annulifera (8%), with 2-45% of infected culicines surviving to adulthood. The parasitic phase of the nematode lasted 5-7 days in all the host species, yielding 1.1-3.2 parasites per II instar and 1.1-2.5 parasites per IV instar. The overall output of parasites per 100 mosquito larvae (infected + uninfected) was highest for Ae. aegypti when mosquitoes were exposed during II instar (2.53 parasites/larva) and for Ar. subalbatus when mosquitoes were exposed during IV instar (1.65/larva), and lowest for Ma. annulifera exposed during IV instar (0.09/larva). For routine laboratory culture of R. iyengari it is convenient to employ Cx. quinquefasciatus as the host yielding 90-190 parasites/100 larva.     

Laboratory tests of the effects of p-cresol and 4-methylcyclohexanol on oviposition by three species of Toxorhynchites mosquitoes

Laboratory experiments tested the effects of p-cresol or 4-methylcyclohexanol at concentrations of 1, 10 and 50 ppm, on oviposition by the mosquitoes Toxorhynchites brevipalpis Theobald, Tx. amboinensis (Doleschall) and Tx. splendens (Wiedemann). A 5 + 5 ppm mixture of the two chemicals was also tested. All three species laid significantly more eggs in cups containing p-cresol, whereas only Tx. brevipalpis and Tx. amboinensis responded similarly to 4-methylcycohexonol and to the mixture of both chemicals. Tx. brevipalpis was, to a relatively limited degree, the most responsive of the three species. Ancillary experiments indicated that the chemicals were acting as attractants, causing more females to fly to treated cups. No stimulant effects were detected either in terms of the proportion of females that initiated oviposition flight (after flying to the cups) or in terms of the number of looping flights executed prior to ejection of an egg.     

Evidences for a stage-specific juvenile hormone binding protein in the hemolymph of the silkworm,Bombyx mori L.: Identification and characterization by photoaffinity labeling and immunological analyses

Two molecular forms of juvenile hormone binding proteins were identified in the larval hemolymph of Bombyx mori by photoaffinity labeling. One form having an Mr of 33 kDa was present constantly in the hemolymph of the third to the fifth instar larvae while the other form having an Mr of 35 kDa was detected in the hemolymph until in the early fifth instar larvae but not in the prewandering larvae and prepupae. A 33 kDa binding protein was purified by hydrophobic interaction chromatography, gel filtration, and native PAGE. Antiserum against 33 kDa binding protein cross-reacted with 35 kDa binding protein on Western blots, suggesting that these binding proteins shared the same epitopes. From the results of saturation binding assays, it was inferred that 33 and 35 kDa binding proteins had a similar binding affinity for JH 1. It was revealed that one of these binding proteins, 35 kDa binding protein, was produced in the fat body in a stage-specific manner: fat body of the early fifth instar larvae synthesized both 33 and 35 kDa binding proteins while that of prewandering larvae synthesized only 33 kDa binding protein. © 1996 Wiley-Liss, Inc.     

The effect of L-canavanine and L-canaline on the hemolymph amino acid composition of the tobacco hornworm,Manduca sexta (L.) (Sphingidae)

Tobacco hornworm larvae, Manduca sexta (L.) (Sphingidae), were administered L-canaline either by parenteral injection or by dietary consumption. The overt toxicity and the alteration of hemolymph amino acids caused by these nonprotein amino acids were evaluated. The LD₅₀ value for parenterally administered canavanine and canaline is 1.0 and 2.5 mg/g fresh body weight, respectively. A dietary concentration of 5.2 mM for canavanine and over 20 mM for canaline represent the respective LC₅₀ values. A large percentage of the larvae reared on diets supplemented with additional arginine, ornithine, or 2,4-diaminobutyric acid in addition to canavanine or canaline were unable to complete larval-pupal ecdysis. These toxic effects were associated with a decreased glutamic acid hemolymph titer and dramatically elevated ornithine. On the other hand, larvae administered canavanine or canaline alone, either by dietary consumption or parenteral injection, experienced less drastic developmental aberrations. These symptoms were in some cases correlated with increased ornithine and glutamic acid titers. Evidence is presented that even a canavanine- and canaline-sensitive insect such as M. sexta has a marked ability to eliminate these protective allelochemicals.     

The effect of whey protein hydrolyzate average molecular weight on the lactic acid fermentation

Summary The batch fermentation of whey permeate to lactic acid was improved by supplementing the broth with enzyme-hydrolyzed whey protein. Hydrolyzates prepared with endoprotease were more stimulatory to acid production rates than were those prepared with exo/endo protease. The effect of hydrolyzate average molecular weight on acid production is presented. Results show that the hydrolyzate having an average molecular weight of 700 is the most stimulatory to acid production rates.     

The effect of sugar meals and body size on fecundity and longevity of female Aedes communis(Diptera: Culicidae)

Abstract. Starved females of Aedes communis (DeGeer) had significantly shorter mean lifespans than females with access to one blood meal and/or sugar meals. For the starving females, lifespan was significantly correlated with wing length. This correlation was not found in sugar- and/or blood-fed females. Fructose and sucrose had similar effects on mosquito lifespan. The 25% and 50% (w/w) solutions of each, increased mean lifespans to 37.8 ± 7.2 and 34.9 ± 9.1 days, respectively. A 10% solution, of either sugar, was significantly inferior, mean lifespan 23.2 ± 10.2 days. The initial sugar meal was very important for female longevity. Subsequent meals aided in ovarian development. One sugar meal, one blood meal, and several sugar meals during oogenesis was found to be the optimal diet for the female mosquitoes in this study.     

The effect of Salinum on the symptoms of dry mouth: a pilot study

The effect of a new saliva substitute. Salinum®, was tested in 37 patients with severe symptoms of reduced salivation. The majority of the patients had suffered from hyposalivation and dry mouth for more than 8 years. The saliva substitute consisted of a water soluble extract of linseed. The physical properties of this extract are similar to those of the glycoproteins of the salivary secretions. The patients used the saliva substitute for a seven days period. Prior to the use of the extract the patients reported that the most severe symptoms of decreased salivation were a feeling of dryness in the mouth and burning sensations in the tongue, pharynx and oesophagus, The majority of the patients reported that the use of Salinum reduced the symptoms of hyposalivation. Great variation in effect occurred from patient to patient. Generally the patients with the most severe symptoms experienced the greatest relief of the symptoms when they used Salinum. Although of short duration the results of this pilot study indicate that an extract of linseeds may compensate for some aspects of the consequences of reduced salivation. Further studies are needed to elucidate the feasibility of the extract as saliva replacement.     

The effect of ectomycorrhizas on the phosphorus and dry weight acquisition of Eucalyptus seedlings

The inoculation ofEucalyptus pilularis seedlings withPisolithus tinctorius and the subsequent development of ectomycorrhizas, led to an improved acquisition of phosphorus (P) from three different sources of P. These sources included insoluble phytate. Dry weight gain of seedlings was increased by inoculation in those treatments where growth was limited by P supply.     

The effect of reversion and duplication of leaf disks on the accuracy of photosynthesis determination by the dry weight method

Experiments with sunflower and fodder cabbage showed that reversion of the leaf disks lower side upwards (towards the radiation source) decreased the photosynthetic rate by as much as 26% both at full (2. 10⁵ erg. cm^?2. sec.^?1) and half density of irradiation. On placing two disks one on another the photosynthetic rate was decreased in the upper disk in a similar way as it was on reversion even if it were placed in the normal position (upper side upwards). But assimilation in the lower disk was only a fraction (0–40%) of that of the upper disk and in thick leaves was within the limits of the compensation point. The sum of the increase in dry weight in both disks due to photosynthesis exceeds the increment in one normally placed disk only in thin leaves and at high density of radiation. On the basis of the differences found it is advised to discard experimental samples where a quarter of the disks were reversed during exposure. On evaluating samples in which some of the disks were duplicated in error, it is most accurate to deduct the weight of one control disk from the weight obtained. The decrease in photosynthetic rate found in reversed and duplicated disks is evidently due to the simultaneous effect of differences in the supply of CO₂ and radiation energy to the assimilating tissue and apparatus and also to differences in the photosynthetic capacity of different leaf tissues.     

Protein partitioning at the isoelectric point: influence of polymer molecular weight and concentration and protein size

We report the partition coefficient, K(p') at the isoelectric point of lysozyme, chymotrypsinogen A, albumin, transferrin, and catalase in 64 different polyethylene(PEG)/ dextran(Dx)/water systems. We study the trends of the partition coefficient with protein type, polymer concentration, and polymer molecular weight. We find that the partition coefficient decreases with increasing tie line length for lysozyme, albumin, transferrin, and catalase for which K(p) is less than 1, but increases for chymotrysinogen for which K(p) is larger than 1. The effect of the tie line length on the partition coefficient is larger for the large proteins than for the small proteins. The partition coefficient decreases with increasing protein molecular weight except for lysozyme suggesting that lysozyme is present as a dimer or a trimer. The partition coefficient decreases with increasing PEG molecular weight, but the magnitude of the increase is larger for the smaller PEG molecular eights and tends to level of at high PEG molecular weight. The partition coefficient increases with increasing dextran (Dx) molecular weight for chymotrypsinogen but decreases for catalase. The partition coefficients of lysozyme, albumin, and transferrin increase with increasing Dx molecular weight from Dx 10(4) to Dx 1.1 x 10(5) and then slightly decrease from Dx 1.1 x 10(5) to Dx 5 x 10(5). The experimental results are analyzed using a statistical thermodynamics model. The experimental results are analyzed using a statistical thermodynamics model. The experiments suggest that protein partitioning at the isoelectric point in aqueous two-phase systems is strongly related to the size of the proteins and polymers. Finally, the impossibility of obtaining data completely independent of polymer concentration is emphasized.     

Accumulation of di-n-butylphthalate in plants and its effect on pigment and protein content

Seedlings of radish ( Raphanus salivas L. cv. Cherry Belle) and wheat ( Triticum aestivum L. cv. Starke II, Weibull) placed in air containing vapour of di- n -butyl-phthalate accumulate phthalate to a concentration that is 10⁶ times that of the surrounding air within 3 days. Particularly high accumulation is found in the cuticular and wax layers. Plants earlier found not to respond to di- n -butylphthalate (wheat) show toxic symptoms (carotenoid and chlorophyll deficiency in strong light, conco-mittant with chloroplast destruction and swollen mitochondria) when the phthalate concentration is further increased by addition to the nutrient medium. Phthalate-affected, white leaves lack the protein moieties normally belonging to the light harvesting complex.     

Allelopathic effect of aqueous extracts of three weed species on the growth and leaf chlorophyll content of bread wheat

The purpose of this study was to evaluate the allelopathic effect of weeds (Avena fatua, Melilotus officinalis and Polypogon hissaricus) on germination, growth, dry biomass and chlorophyll concentration of three cultivars of wheat (Ata Habib, Pirsabaq and Serin). In germination test, different concentrations of aqueous extracts (5, 10 and 15?g/l) of the three weeds significantly reduced percent germination; however, 15?g/l extract of M. officinalis resulted in complete failure of germination of cultivar Pirsabaq. In pot culture, root and shoot length, chlorophyll concentration and seedling dry biomass of the three wheat varieties showed differential responses to different weeds. Aqueous extract at 15?g/l of A. fatua increased root and shoot length and dry biomass of cultivar Pirsabaq; however, these parameters were significantly retarded in other two wheat cultivars by extract of weeds. Moisture content of the cultivars did not show any response to allelopathic stress of the weeds. In contrast, chlorophyll concentration in Pirsabaq and Serin was significantly increased by aqueous extract of all the weeds but reduced it in cultivar Ata Habib by 50%. In general, Ata Habib was found to be the most sensitive cultivar to the imposed allelopathic stress. The phytotoxic potential of three weeds was found in the order of A. fatua?>?M. officinalis?>?P. hissaricus.     

Yolk protein in the pharaoh's ant: Influence of larvae and workers on vitellogenin and vitellin content in queens

Summary Vitellin from the pharaoh's ant,Monomorium pharaonis (L.), was found to contain a single apoprotein with aM_r of 198.3 kDa. The protein is a glycoprotein exposing mannose containing carbohydrate groups.Antibodies to pharaoh's ant vitellin (v_t), raised in rabbits, were used to determine the head-, thorax-, and abdominal contents of vitellogenin and vitellin (v_g+v_t) in queens in relation to the presence or absence of larvae and workers using an ELISA test. Thev_g+v_t contents were also compared to the egg-laying rate and the weight of the queens.The results confirm the existence of a positive correlation betweenv_g+v_t contents in the queens and their access to larvae, probably related to the queens' preferential feeding on larval secretions. In queens without larvae the abdominalv_g+v_t contents declined in concordance with a low oviposition rate of 6–8 eggs/day. In spite of cessation of egg-laying within 24 hours after removal of both larvae and workers, the queens maintained basal contents ofv_g+vt. This may indicate that the presence of larvae is not only essential for the nutrition of the queens, but also for the uptake of vitellogenin in the growing oocytes. This additional stimultive factor may be based on the queens' response to primer pheromones liberated by the larvae.v_g+v_t could not be demonstrated in workers or larvae with the ELISA test. If anyv_g+vt is present in larvae and workers the amount is lower than the detection limit (1–2 ng/individual). This seems to rule out the possibility of transfer of proteins of this kind to the queens.