The application of gas liquid chromatography to the simultaneous analysis of sugars and sugar phosphates in biological samples /|nby David J. LeBlanc. -- 260 St. Catharines [Ont. : s. n.],

A study was undertaken' to determine the applicability of gas liquid chromatography to the simultaneous analysis of sugars and sugar phosphates from biological samples. A new method of silylation involving dimethylsulfoxide, hexamethyldisilazane, trimethylchlorosilane and cyclohexane (1:0.2:0.1:1) which rapidly silylated sugars and sugar phosphates was developed. Subsequent chromatography on a 5% SE-52 column gave good resolution of the sugar and sugar phosphate samples. Sugar phosphates decomposed during chromatography and were lost at the 7 x 10-3 ~mole level. Acidic ethanol extraction of yeast samples revealed background contamination from the yeast sample, the culture medium and the silylation reagents which would further limit the level of detection obtainable with the glc for sugars in biological samples to the 3 x 10-4 ~mole level.

An investigation into the effects of homopteran honeydew sugars versus floral nectar sugars on black fly longevity, flight performance and digestion /

Floral nectar is thought to be the primary carbohydrate source for most dipteran species. However, it has been shown that black flies (Burgin & Hunter 1997 a,b,c), mosquitoes (Foster 1995; Burkett et al. 1999; Russell & Hunter 2002), deer flies (Magnarelli & Burger 1984; Janzen & Hunter 1998; Ossowski & Hunter 2000), horse flies (Schutz & Gaugler 1989; Hunter & Ossowski 1999) and sand flies (MacVicker et al. 1990; Wallbanks et al. 1990; Cameron et al. 1992, 1995; Schlein & Jacobson 1994, 1999; Hamilton & EI Naiem 2000) feed on homopteran honeydew as well as floral nectar. Prior to 1997 floral nectar was thought to be the main source of carbohydrates for black flies. However, Burgin & Hunter (1 997a) demonstrated that up to 35% of black flies had recently consumed meals of homo pte ran honeydew. This information has necessitated a re-assessment of many life history aspects of black flies. Attempts are being made to examine the effects of nectar versus honeydew on black fly fecundity and parasite transmission (Hazzard 2003). Recently, Stanfield and Hunter (unpublished data) have shown that in female black flies, honeydew sugars produce flights of longer distance and duration than do nectar sugars. This thesis examines two aspects of black fly biology as it relates to sugar meal consumption. First, the effects of honeydew and nectar on black fly longevity are examined. Second, the proximate causation behind longer flight performances in honeydew-fed flies will be examined. The comparison between these two sources is important because nectar is composed of mainly simple sugars (monosaccharides and disaccharides) whereas honeydew is composed of both simple and complex sugars (including trisaccharides and tetrasaccharides ).

An investigative study on the effects of black fly (diptera: simuliidae) sugar meals on reproductive success and parasite transmission /

Black flies are opportunistic sugar-feeders. They take sugar meals from Homopteran honeydew secretions or plant nectars, depending on availability. Homopteran honeydew secretions contain both simple and complex carbohydrates while plant nectars contain primarily simple carbohydrates. In order to determine whether honeydew secretions offer more energy than plant nectars to their insect visitors a study of wild-caught black flies was undertaken in Algonquin Provincial Park, Canada during the spring of 1 998 and 1 999. It was hypothesized that female black flies maintained on honeydew sugars will survive longer, produce more eggs and have a greater parasite vectoring potential than those maintained on artificial nectar or distilled water. Results demonstrated that: (1) host-seeking female Prosimulimfuscum/mixtum and Simulium venustum maintained on artificial honeydew did not survive longer than those maintained on artificial nectar when fed ad libitum; (2) fiiUy engorged S. venustum and Simulium rugglesi maintained on artificial honeydew did not produce more eggs than those maintained on artificial nectar when fed ad libitum; and (3) S. rugglesi did not have a greater vectoring potential of Leucocytozoon simondi when maintained on artificial honeydew as opposed to artificial nectar when fed ad libitum. However, all flies maintained on the two sugars (artificial honeydew and artificial nectar) survived longer, produce more eggs and had greater vectoring potential than those maintained on distilled water alone.

Timing of di-ammonium phosphate addition to fermenting chardonnay must : effect on nitrogen utilization, ethyl-carbamate formation, and CAR1 expression by yeast /

The maximum amount of ethyl carbamate (EC), a known animal carcinogen produced by the reaction of urea and ethanol, allowed in alcoholic beverages is regulated by legislation in many countries. Wine yeast produce urea by the metabolism of arginine, the predominant assimilable amino acid in must. This action is due to arginase (encoded by CARl). Regulation of CARl, and other genes in this pathway, is often attributed to a well-documented phenomenon known as nitrogen catabolite repression. The effect of the timing of di-ammonium phosphate (DAP) additions on the nitrogen utilization, regulation of CARl, and EC production was investigated. A correlation was found between the timing of DAP addition and the utilization of nitrogen. When DAP was added earlier in the fermentations, less amino nitrogen and more ammonia nitrogen was sequestered from the media by the cells. It was also seen that early DAP addition led to more total nitrogen being used, with a maximal difference of ~25% between fermentations where no DAP was added versus addition at the start of the fermentation. The effect of the timing ofDAP addition on the expression of CARJ during fermentation was analyzed via northern transfer and the relative levels of CARl expression were determined. The trends in expression can be correlated to the nitrogen data and be used to partially explain differences in EC formation between the treatments. EC was quantified at the end of fermentation by GC/MS. In Montrachet yeast, a significant positive correlation was found between the timing of DAP addition, from early to late, and the final EC concentration m the wine (r = 0.9226). In one of the fermentations, EC levels of 30.5 ppb was foimd when DAP was added at the onset of fermentation. A twofold increase (69.5 ppb) was observed when DAP was added after 75% of the sugars were metabolized. When no DAP was added, the ethyl carbamate levels are comparable at a value of 38 ppb. In contrast, the timing of DAP additions do not affect the level EC produced by the yeast ECU 18 in this manner. The study of additional yeast strains shows that the effect of DAP addition to fermentations is strain dependent. Our results reveal the potential importance of the timing of DAP addition to grape must with respect to EC production, and the regulatory effect of DAP additions on the expression of genes in the pathway for arginine metabolism in certain wine yeast strains.

Host-parasite relations in a mycoparasitic system : alterations in membrane permeability and lipid composition in Choanephora cucurbitarum infected by Piptocephalis virginiana /

examined in Choanephora cucurbita rum during the early stages of infection by Piptocephalis virginiana » There was a small but consistent increase in the leakage of electrolytes, amino acids and sugars as a result of infection. These low levels of differential leakage in infected tissues are explained on the basis of the nature of this obligate, biotrophic, mycoparasitic system. Quantitative analysis of the twenty six amino acids and amino compounds detected in the leacheates — showed similar profiles in infected and control host and no new species of amino acids or amino compounds were detected in either infected or control host leacheates. Comparatively high amounts of aspartic acid, glutamic acid and alanine were found in the leacheates of host and infected host . Analyses of the sugars comprising the leacheates of infected and control host showed the presence of eight sugars, among which glucose was found in significant amounts (50-53%) ' The nutritional implication of this preferential leakage is discussed. No significant difference was observed in the leacheates of infected host sugar profiles compared with that of the control host. Profiles of the internal pool sugars of infected and control host did not reflect that obtained from the leacheate data, perhaps owing to leakage of sugars in a selective manner . Membrane lipid analyses yielded higher levels of lipid in infected host compared with the control, both at the 24 h and 36 h analyses. In addition, preliminary investigations of phosphorous-32 incorporation and turnover in phospholipids showed higher levels of 32p incorporation and turnover in infected host compared with the control. No apparent difference was noted in the profiles of the neutral lipid classes and the polar lipid classes of the membrane lipids as determined by one and two dimensional thin-layer chromatography respectively. However, a small but consistently higher degree of unsaturation was detected in the fatty acids of infected tissue compared with the control. Also, '^''-^^''^^'-'-^'^^c acid, a polyunsaturated fatty acid previously reported to show a direct correlation during the early stages of infection and the degree of parasitism of P. virginiana on C. cucurbitarum , was found in higher amounts in infected host membrane lipids compared with that of the control host. The implications of these membrane lipid alterations are discussed with particular reference to the small but consistently higher leakage of electrolytes, amino acids and sugars observed during infection in this study.

An investigation of sugar feeding by black flies (Diptera: Simuliidae)

Although much research has been conducted on blood-meal acquisition in adult female black flies (Diptera: Simuliidae), the same cannot be said for sugarmeals. Both sexes feed on sugar which provides energy for flight and it has been commonly held that nectar is the major carbohydrate source. This thesis addresses the question of whether a non-floral carbohydrate source, specifically homopteran honeydew, is ingested by male and female black flies. Black flies reared in the laboratory have been observed to readily ingest freshly excreted and older (dry) honeydew when presented with honeydew coated tamarack branches. Field work was conducted in Algonquin Park, Ontario in the spring and summer of 1993. Three separate studies were designed to test whether homopteran honeydew is an important carbohydrate source for black flies and whether flies from different habitats utilize different sugar sources. The sugars melezitose and / or stachyose are known to occur in a variety of homopteran honeydews and therefore were used as indicators of honeydew feeding by black flies. In the first study, black flies were collected with insect nets from a stand of Larix larcina heavily infested with honeydew - producing homopterans (Adelges lariciatus). Six black fly species were captured: Simulium venustum, S. rostra tum, S. vittatum, Stegopterna mutata, S. aureum and S. quebecense. Samples of honeydew and individual black flies were tested using thin layer chromatography (T. L. C.) with fructose, glucose, sucrose, turanose, melezitose, raffinose and stachyose as standards. All sugars except turanose and melezitose were found in the adelgid honeydew samples. Since the sugar melezitose was absent from ~ honeydew samples, stachyose was used to indicate that black flies were feeding from this particular honeydew source. Of the 201 black flies tested, 194 contained sugars which occurred in 16 combinations. Stachyose combinations excluding melezitose, present in 45.9 % of flies, were used to indicate that black flies had been feeding on the adelgid honeydew. In the second study, black flies were collected in the morning and evening on 8 collection dates, using a vehicle mounted insect net. The crops and midguts of 10 male and 10 female Simulium venustum were dissected on each sample date. In total the gut contents of 320 individual flies were analysed by T. L. C. The sugars identified from these flies were present in the following proportions: fructose (100.0%), glucose (100.0%), sucrose/turanose (50.4%), melezitose (30.3%), raffinose (18.8%) and stachyose (8.7%). These sugars occurred in fourteen different combinations. It is argued that the presence of melezitose and / or stachyose indicates that black flies had fed on homopteran honeydew. Significantly more female flies (40.0%) than male flies (27.5%) had fed on honeydew. In the third study, adult black flies were sampled by sweep netting vegetation in four habitats in the morning and evening on 8 collection dates. The habitats are as follows: (1) Davies Bog, (2) Abandoned Air Field (dominated by blueberries, Vaccinium spp.), (3) Deciduous Habitat and (4) Coniferous Habitat. Sugars in the crops and midguts of female flies were tested by T. L. C. and, for S. venustum, it was found that significantly fewer flies (18.8%) from the Air Field contained honeydew than from the other three sites (Davies Bog, 34.4%; Deciduous Habitat, 36.2%; Coniferous Habitat, 25.0%). Of the 1287 black flies tested individually by T. L. C. 441 (34.3%) contained melezitose and / or stachyose sugars indicating that this proportion of the population were feeding from Homopteran honeydew. It is therefore clear that floral (nectar) sugars are not the only source of carbohydrates available to black flies.

Glucosamine resistance in the yeast, Saccharomyces cerivisae [sic]

By using glucosamine resistant mutants of Saccharomyces ceriv~sa~ an attempt was made to discover the mechanisms which cause glucose repression and/or the Crabtree effect. The strains used are 4B2, GR6, lOP3r, GR8l and GRI08. 4B2 is a wild type yeast while the others are its mutants. To characterize the biochemical reactions which made these mutants resistant to glucosamine poisoning the following experiments were done~ 1. growth and respiration; 2. transport of sugars; 3. effect of inorganic phosphate (Pi): 4. Hexokinase; 5. In yivo phosphorylation. From the above experiments the following conclusions may be drawn: (i) GR6 and lOP3r have normal respiratory and fermentative pathways. These mutants are resistant to glucosamine poisoning due to a slow rate of sugar transport which is due to change in the cell membrane. (ii) GR8l has a normal respiratory pathway. The slow growth on fermentable carbon sourCEE indicates that in GR8l the lesion is in or associated with the glycolytic pathway. The lower rate of sugar transport may be due to a change in energy metabolism. The invivo phosphorylation rate indicates that in GR81 facilitated diffusion is the dominant transport mechanism. (iii) GR108 msa normal glycolytic pathway but the respiratory pathway is abnormal. The slow rate of sugar transport is due to a change in energy metabolism. The lower percentage of in vivo phosphorylation is probably due to a lowered availability of ATP because of the mitochondrial lesion. In all mutants resistance to glucosamine poisoning is due to a lower rate of utilization of ATP. which is caused by various mechanisms (see above), making less ADP available for phosphorylation via ATP synthase which utilizes inorganic phosphate. Because of the lower utilization of Pi, the concentration of intra-mitochondrial Pi does not go down thus protecting mutants from glucosamine poisoning.

Isolation and partial characterization of host cell wall surface glycoproteins: their involvement in agglutination, attachment and appressorium formation by piptocephalis virginiana

Cell surface proteins obtained by alkaline extraction from isolated cell walls of Mortierella pusilla and M. candelabrum, host and nonhost, respectively, to the mycoparasite, Piptocephalis virginiana, were tested for their ability to agglutinate mycoparasite spores. The host cell wall protein extract had a high agglutinating activity (788 a.u. mg- t ) as compared with the nonhost extract (21 a.li. mg- t ). SDS-polyacrylamide gel electrophoresis of the cell wall proteins revealed four protein bands, a, b, c, and d (Mr 117, 100, 85 and 64 kd, respectively) at the host surface, but not at the nonhost surface, except for the faint band c. Deletion of proteins b or c from the host cell wall protein extract significantly reduced its agglutinating activity. Proteins band c, obtained as purified preparations by a series of procedures, were shown to be two glycoproteins. Carbohydrate analysis by gas chromatography demonstrated that glucose and Nacetylglucosamine were the major carbohydrate components of the glycoproteins. It was further shown that the agglutinating activity of the pure preparation containing both band c was 500-850 times that of the single glycoproteins, suggesting the involvement of both glycoproteins in agglutination. The results suggest that the glycoproteins band c are the two subunits of agglutinin present at the host cell surface. The two glycoproteins band c purified from the host cell wall protein extract were further examined after various treatments for their possible role in agglutination, attachment and appressorium formation by the mycoparasite. Results obtained by agglutination and attachment tests showed: (1) the two glycoprotein-s are not only an agglutinin responsible for the mycoparasite spore agglutination, but may also serve as a receptor for the specific recognition, attachment and appressorium formation by the mycoparasite; (2) treatment of the rnycoparasite spores with various sugars revealed that arabinose, glucose and N-acetylglucosamine inhibited the agglutination and attachment activity of the glycoproteins, however, the relative percentage of appressorium formation was not affected by the above sugars; (3) the two glycoproteins are relatively stable with respect to their agglutinin and receptor functions. The present results suggest that the agglutination and attachment may be mediated directly by certain sugars present at the host and mycoparasite cell surfaces while the appressorlum formation may be the response of complementary combinations of both sugar and protein, the two parts of the glycoproteins at the interacting surfaces of two fungi.

Probing cell surfaces of host and nonhost species of a mycoparasite, using FITC-labeled lectins

Cell surfaces of susceptible host species (Mortierella pusllla and Cboanepilora cucurbitarum ), resistant host (Pilascolomyces articulosus ), nonhost (Mortierella candelabrum ) and the mycoparasite (Piptocepilalis virginiana) were examined for sugar distribution patterns using light and fluorescent microscopy techniques. The susceptible host, resistant host and the mycoparasite species exhibited a similar sugar distribution profile; they all showed N-acetyl glucosamine and D-glucose on their cell wall surfaces. The nonhost cell wall surface showed a positive binding reaction to FITClectins specific for N-acetyl glucosamine and also for OI.-fucose, N-acetyl galactosamine and galactose. Treatment of these fungi with mild concentrations of proteinases (both commercial as well as the mycoparasiteproteinase) resulted in the revelation of additional sugars on the fungal cell walls. The susceptible host treated with proteinase expressed higher levels of N-acetyl glucosamine and D-glucose. The susceptible host also showed the presence of OI.-fucose, N-acetyl galactosamine and galactose. The proteinasetreated susceptible host cell walls also showed an increase in the levels of attachment with the mycoparasite. Treatment of the resistant host with proteinases revealed OI.-fucose in addition to N-acetyl glucosamine and D-glucose. Treatment of the nonhost cell wall with proteinase resulted in the exposure of low levels of D-glucose, in addition to sugars found on the untreated nonhost cell wall surface. The mycoparasite treated with proteinase revealed OI.-fucose, N-acetyl galactosamine and galactose on its cell surface in addition to the sugars N-acetyl glucosamine and D-glucose. Protoplasts were isolated from hosts and nonhost fungi and their surfaces were examined for sugar distribution patterns. The susceptible host and nonhost protoplast membranes showed all the sugars (N-acetyl glucosamine, D-glucose, (It.-fucose, N-acetyl galactosamine and galactose) tested for. The resistant host protoplast membrane however, had only N-acetyl glucosamine and D-glucose exposed. This sugar distribution profile resembles that exhibited by the untreated resistant host cell wall, as well as that shown by the untreated mycoparasite cell surface. Only susceptible host protoplasts were successful in attaching to the mycoparasite surface. Resistant host protoplasts did not show any interaction with the i mycoparasite cell surface. Both susceptible as well as resistant host protoplasts were incapable of attaching to agarose beads surface-coated with specific carbohydrates. The mycoparasite however, did attach to agarose beads surface-coated with either N-acetyl glucosamine, D-glucose/Dmannose or o:,- methyl-D-mannose. The relevance of the cell wall and the protoplast membrane in the light of the present results, in reacting appropriately to bring about either a susceptible, a resistant or a nonhost response has been discussed.

The effect of age on the structure and compostition of the cell walls of Choanephora cucurbitarum

The effect of age on the structure and composition of isolated and purified cell walls from cultures of Choanephora cucurbitarum was investigated by microchemical analyses, visible and infrared spectrophotometry, x-ray diffractometry and electron microscopy. Qualitative evaluation revealed the presence of lipids, proteins, neutral sugars, strong alkali soluble sugars, chitin, chitosan and uronic acids in the cell walls of both the 1 and 7 day old cultures. As the mycelium aged, there was a slight but statistically significant increase in the protein content, and a pronounced rise in the chitin and neutral sugar constituents of the cell walls. Conversely, the decrease in the chitosan content during this period had the net effect of altering the chitin: chitosan ratio from near unity in the younger cultures, to a 2:1 ratio in the 7 day old cell wall samples. Glutaraldehyde-osmium fixed thin sections of the 1 day old vegetative hyphae of £. curbitarum revealed the presence of a monolayered cell wall, which upon aging became bilayered. Replicas of acid hydrolysed cell walls demonstrated that both the 1 and 7 day old samples possessed an outer layer which was composed of finely granular amorphous material and randomly distributed microfibrils. The deposition of an inner secondary layer composed of parallel oriented microfibrils in the older hypha was correlated with an increase in the chitin content in the cell wall. The significance of these results with respect to the intimate relationship between composition and structure is discussed.

The regulation of oat coleoptile phosphoenolpyruvate carboxylase and malic enzyme by Hâ ½ and metabolites : kinetic evidence for and against a cytosolic pH-stat

Phosphoenolpyruvate carboxylase (PEPC) and malic enzyme activities in soluble protein extracts of Avena coleoptiles were investigated to determine whether their kinetics were consistent with a role in cytosol pH regulation. Malic enzyme activity was specific for NADP+ and Mn2+. Maximal labelled product formation from [14C]-substrates required the presence of all coenzymes, cofactors and substrates. Plots of rate versus malate concentration, and linear transformations there- 2 of, indicated typical Michaelis-Menten kinetics at non-saturating malate levels and substrate inhibition at higher malate levels. pH increases between 6.5 and 7.25 increased near-optimal activity, decreased the degree of substrate inhibition and the Kmapp(Mn2+) but did not affect the Vmax or Kmapp(malate). Transformed data of PEPC activity demonstrated non-linear plots indicative of non-Michaelian kinetics. pH increases between 7.0 and 7.6 increased the Vmax and decreased the Km app (Mg2+) but did not affect the Kmapp(PEP). Various carboxylic acids and phosphorylated sugars inhibited PEPC and malic enzyme activities, and these effects decreased with pH increases. Metabolite inhibited malic enzyme activity was non-competitive and resulted mainly from Mn2+ chelation. In contrast, metabolite inhibited PEPC activity was unique for each compound tested, being variously dependent on the PEP concentration and the pH employed. These results indicate that fluctuations in pH and metabolite levels affect PEPC and malic enzyme activities similarly and that 3 the in vitro properties of PEPC are consistent with its proposed role in a pH-stat, whereas the in vitro properties of the malic enzyme cannot be interpreted in terms of a role in pH regulation.

Investigations on the host-parasite interface of a mycoparasite system /

The cell wall composition of Choanephora cucur - bitarum and the host-parasite interface, after infection with Piptocephalis virginiana , were examined in detail. The cell walls of C_. cucurbitarum were determined to be composed of chitin (17%), chitosan (28.4%), neutral sugars (7.2%),uronic acid (2.4%), proteins (8.2%) and lipids (13.8%). The structure of hyphal walls investigated by electron microscopy of shadowed replicas before and after alkali-acid hydrolysis, showed two distinct regions: microfibrillar and amorphous. The microfibrils which were composed of mainly chitin, were organized into two distinct layers: an outer, thicker layer of randomly orientated microfibrils and an inner, thin layer of parallel microfibrils.Electronmicrographs of the host-parasite interface of C_. cucurbitarum and the mycoparasite , P_. virginiana , 30 h following inoculation, showed that the sheath zone has a similar electron density to that of the host cell wall. The sheath was not present around the young (18 h old) haustorium. High-resolution autoradiographs of infected host hyphae showed that radioactive N-acetyl-D-glucosamine , a precursor of chitin, was incorporated preferentially in the host cell wall and sheath zone. Cell fractionation of label fed hyphae showed that 84% of the label was present in the cell wall and specifically in the chitin portion of the wall. The antifungal antibiotic, Polyoxin D, a specific inhibitor of the enzyme, chitin synthetase, suppressed the incorporation of the label in the cell wall and sheath zone and resulted in a decrease in electron density of the developing sheath. The significance of these results is discussed in the light of host resistance.

Chemoenzymatic synthesis of amaryllidaceae alkaloids and their C-1 analogues : symmetry based approach to total synthesis of thebaine

Described herein is the chemoenzymatic total synthesis of several Amaryllidaceae constituents and their unnatural C-I analogues. A new approach to pancratistatin and related compounds will be discussed along with the completed total synthesis of 7 -deoxypancratistatin and trans-dihydrolycoricidine. Evaluation of all new C-l analogues as cancer cell growth inhibitory agents is described. The enzymatic oxidation of dibromobenzenes by Escherichia coli 1M 109 (pDTG60 1) is presented along with conversion of their metabolites to (-)-conduritol E. Investigation into the steric and functional factors governing the enzymatic dihydroxylation of various benzoates by the same organism is also discussed. The synthetic utility of these metabolites is demonstrated through their conversion to pseudo-sugars, aminocyclitols, and complex bicyclic ring systems. The current work on the total synthesis of some morphine alkaloids is also presented. Highlighted will be the synthesis of several model systems related to the efficient total synthesis of thebaine.

The Influence of Male Diet on Life History Traits of Female Mosquitoes

In the field, mosquitoes characteristically feed on sugars soon after emergence and intermittently during their adult lives. Sugar meals are commonly derived from plant nectar and homopteran honeydew, and without them, adults can only survive for a few days on larval reserves. In addition to sugar, females of most species rely on blood for the initiation and maintenance of egg development; thus their reproductive success depends to some extent on the availability of blood hosts. Males, on the other hand, feed exclusively on sugars. Consequently, their sexual maturation and reproductive success is largely dependent upon access to sugar sources. Plant nectar and homopteran honeydew are the two main sugar sources utilized by mosquitoes in the wild. Previous laboratory studies had shown that differences between nectar sources can affect the survivorship and biting frequency of disease vectoring mosquitoes. However, little is known on how sugar composition influence the reproductive processes in male mosquitoes. Male mosquitoes transfer accessory gland proteins and other hormones to their mates along with sperm during mating. In the female, these seminal fluid constituents exert their influence on reproductive genes that control ovulation and vitellogenesis. The present study tests the hypothesis that the mates of males consuming different sugar meals will exhibit varying levels of induction of vitellogenin (a gene which regulates the expression of egg yolk precursor proteins). Real-time quantitative RT-PCR was used to investigate how each sugar meal indirectly influences vitellogenin mRNA abundance in female Anopheles stephensi following mating. Results indicate that mates of nectar-fed males exhibit 2-fold greater change in vitellogenin expression than the mates of honeydew-fed males. However, this response did not occur in non-blood fed controls. These findings suggest that the stimulatory effect of mating on vitellogenesis in blood meal-reliant (i.e. anautogenous) mosquitoes may only be synergistic in nature. The present study also sought to compare the potential fitness costs of mating incurred by females that do not necessarily require a blood meal to initiate a reproductive cycle (i.e., exhibit autogeny). Females of the facultatively autogenous mosquito, Culex molestus were allowed to mate with males sustained on either nectar or honedyew. Mean lifetime fecundity and survivorship of females under the two different mating regimes were then recorded. Additionally, one-dimensional gel electrophoresis was used to verify the transfer of male accessory gland proteins to the sperm storage organs of females during mating.While there was no significant difference in survival between the test treatments, the mates of nectar-fed males produced 11% more eggs on average than mates of honeydew-fed males. However, additional data are needed to justify the extrapolation of these findings to natural settings. These findings prompt further investigation as the differences caused by diet variation in males may be reflected across other life history traits such as mating frequency and insemination capacity.