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Biomedical subjects

A E Douglas

Publications and source records attributed to A E Douglas.

At least 19 recordsLinked to original sources
PubMed2006

Phloem-sap feeding by animals: problems and solutions.

The incidence of phloem sap feeding by animals appears paradoxical. Although phloem sap is nutrient-rich compared with many other plant products and generally lacking in toxins and feeding deterrents, it is consumed as the dominant or sole diet by a very restricted range of animals, exclusively insects of the order Hemiptera. These insects display two sets of adaptations. First, linked to the high ratio of non-essential:essential amino acids in phloem sap, these insects contain symbiotic micro-organisms which provide them with essential amino acids. For example, bacteria of the genus Buchnera contribute up to 90% of the essential amino acids required by the pea aphid Acyrthosiphon pisum feeding on Vicia faba. Second, the insect tolerance of the very high sugar content and osmotic pressure of phloem sap is promoted by their possession in the gut of sucrase-transglucosidase activity, which transforms excess ingested sugar into long-chain oligosaccharides voided via honeydew. Various other animals consume phloem sap by proxy, through feeding on the honeydew of phloem-feeding hemipterans. Honeydew is physiologically less extreme than phloem sap, with a higher essential:non-essential amino acid ratio and lower osmotic pressure. Even so, ant species strongly dependent on honeydew as food may benefit from nutrients derived from their symbiotic bacteria Blochmannia.

Amino Acids↗
PubMed2006

Sweet problems: insect traits defining the limits to dietary sugar utilisation by the pea aphid, Acyrthosiphon pisum.

Plant phloem sap is an extreme diet for animals, partly because of its high and variable sugar content. The physiological and feeding traits of the pea aphid Acyrthosiphon pisum that define the upper and lower limits to the range of dietary sucrose concentrations utilised by this insect were determined principally using chemically defined diets containing 0.125-1.5 mol l(-1) sucrose. On the diets with 0.125 mol l(-1) and 1.5 mol l(-1) sucrose, the aphids died as larvae within 8 and 14 days of birth, respectively. On the other diets, 60-96% of aphids developed to adulthood, and the 0.5 mol l(-1) and 0.75 mol l(-1) diets supported the highest fecundity. The diet with 0.125 mol l(-1) sucrose was ingested at 36% of the rate of the 0.25 mol l(-1) sucrose diet, but >90% of ingested sucrose-carbon was assimilated on both diets. This suggests that the lower limit is dictated by the aphid feeding response, specifically, a requirement for a minimal concentration of sucrose for sustained feeding. The haemolymph osmotic pressure of aphids on diets with 0.125-1.5 mol l(-1) sucrose was up to 68% higher than on 0.125-1.0 mol l(-1) sucrose diets, but diet consumption and sucrose-carbon assimilation was not reduced on the very high sucrose diets relative to 1.0 mol l(-1) sucrose. This suggests that failure of the osmoregulatory capacity of the insects on high sucrose diets may define the upper limit to the range of dietary sucrose utilised by the aphids. The mean haemolymph osmotic pressure of aphids on plants with phloem sap containing 0.37-0.97 mol l(-1) sucrose was 1.61+/-0.063 MPa (mean +/- s.e.m.), not significantly different from that (1.47+/-0.059 MPa) on diets with 0.25-1.0 mol l(-1) sucrose. It is concluded that the osmoregulatory response of aphids to diets and plants are comparable, and, more generally, that the feeding and osmoregulatory capabilities of the aphids are compatible with the phloem sugar levels commonly encountered by aphids feeding on plants.

Animals↗
PubMed2005

The significance of gut sucrase activity for osmoregulation in the pea aphid, Acyrthosiphon pisum.

The osmotic pressure of the body fluids of aphids is lower than in their diet of plant phloem sap. It is hypothesised that aphids reduce the osmotic pressure of ingested food by sucrase-mediated hydrolysis of dietary sucrose to glucose and fructose, and the polymerisation of glucose into oligosaccharides of low osmotic pressure per hexose unit. To test this hypothesis, the impact of the alpha-glucosidase inhibitor acarbose on the sugar relations and osmoregulation of aphids was explored. Acarbose inhibited sucrase activity in gut homogenates and the production of monosaccharides and oligosaccharides in the honeydew of live aphids. Acarbose caused an increase in the haemolymph osmotic pressure for aphids reared on a diet (containing 0.75 M sucrose) hyperosmotic to the haemolymph and not on the isoosmotic diet containing 0.2 M sucrose. It did not affect aphid feeding rate over 2 days, except at high concentrations on 0.75 M sucrose diet, and this may have been a secondary consequence of osmotic dysfunction. Acarbose-treated aphids died prematurely. With 5 microM dietary acarbose, mean survivorship on 0.2 M sucrose diet was 4.2 days, not significantly different from starved aphids, indicating that, although these aphids fed, they were deprived of utilisable carbon; and on 0.75 M sucrose diet, mean survivorship was just 2.8 days, probably as a consequence of osmotic failure. It is concluded that the aphid gut sucrase activity is essential for osmoregulation of aphids ingesting food hyperosmotic to their body fluids.

Acarbose↗
PubMed2005

Aphid-symbiotic bacteria cultured in insect cell lines.

The cells and tissues of many aphids contain bacteria known as "secondary symbionts," which under specific environmental circumstances may be beneficial to the host insect. Such symbiotic bacteria are traditionally described as intractable to cultivation in vitro. Here we show that two types of aphid secondary symbionts, known informally as T type and U type, can be cultured and maintained in three insect cell lines. The identities of the cultured bacteria were confirmed by PCR with sequencing of 16S rRNA gene fragments and fluorescence in situ hybridization. In cell lines infected with bacteria derived from aphids harboring both T type and U type, the U type persisted, while the T type was lost. We suggest that the two bacteria persist in aphids because competition between them is limited by differences in tropism for insect tissues or cell types. The culture of these bacteria in insect cell lines provides a new and unique research opportunity, offering a source of unibacterial material for genomic studies and a model system to investigate the interactions between animal cells and bacteria. We propose the provisional taxon names "Candidatus Consessoris aphidicola" for T type and "Candidatus Adiaceo aphidicola" for U type.

Animals↗
PubMed2003

Coral bleaching--how and why?

Bleaching refers to the loss of colour in symbioses between dinoflagellate algae of the genus Symbiodinium and marine benthic animals, e.g. corals. Bleaching generally results in depressed growth and increased mortality, and it can be considered as a deleterious physiological response or ailment. An explanatory framework for the causes of bleaching comprises three elements: the external factors or triggers of bleaching, e.g. elevated temperature; the symptoms, including elimination of algal cells and loss of algal pigment; and the mechanisms, which define the response of the symbiosis to the triggers, resulting in the observed symptoms. The extent to which bleaching in different symbioses and in response to different triggers involves common mechanisms is currently unknown, but a contribution of interactions between the algal and animal partners to bleaching is predicted. Symbioses vary in their susceptibility to bleaching as a result of genetic variation in Symbiodinium and acclimatory responses of the animal. The evolutionary explanation for bleaching is obscure. Perhaps, bleaching was of selective advantage to the animal hosts under different (more benign?) environmental conditions than the present, or bleaching may be a negative by-product of an otherwise advantageous symbiotic trait, such as the elimination of damaged algal cells.

Animals↗
PubMed2003

The causes and processes of the mid-summer population crash of the potato aphids Macrosiphum euphorbiae and Myzus persicae (Hemiptera: Aphididae).

Populations of many phloem-feeding aphid species in temperate regions increase exponentially in early summer and then 'disappear', usually over a time-scale of a few days, in July. To understand these dynamics, empirical investigation of the causes and modelling of the processes underlying population change are required. Numbers of the aphids Myzus persicae(Sulzer) and Macrosiphum euphorbiae (Thomas), monitored over three years in commercial potato fields in the UK, increased to a maximum of 2-2.5 per leaflet on 16 July in 1999 and 2001, and then declined to < 0.25 per leaflet by 26 July. In 2000, aphid numbers remained very low (< 0.25 per leaflet) throughout the season. The onset of the crash in aphid numbers (16-19 July in 1999 and 2001) was consistently associated with changes in the phloem amino acid composition of potato leaflets. Natural enemies, including syrphids, parasitoids, coccinellids, chrysopids and entomopathogenic fungi, increased in abundance throughout the sampling period. The incidence of winged emigrant aphids prior to the crash was low (< 10%). Experimental manipulation during 2001 demonstrated that, during the crash period, the fecundity of aphids (caged on leaves to exclude natural enemies) was depressed by 25-45% relative to earlier in the season, and that presence of natural enemies reduced aphid numbers by up to 68%. Using these data, an excitable medium model was constructed, which provided a robust description of aphid population dynamics in terms of plant development-induced changes in aphid fecundity and temporal change in natural enemy pressure.

Amino Acids↗
PubMed2003

Diversity of bacteria associated with natural aphid populations.

The bacterial communities of aphids were investigated by terminal restriction fragment length polymorphism and denaturing gradient gel electrophoresis analysis of 16S rRNA gene fragments generated by PCR with general eubacterial primers. By both methods, the gamma-proteobacterium Buchnera was detected in laboratory cultures of six parthenogenetic lines of the pea aphid Acyrthosiphon pisum and one line of the black bean aphid Aphis fabae, and one or more of four previously described bacterial taxa were also detected in all aphid lines except one of A. pisum. These latter bacteria, collectively known as secondary symbionts or accessory bacteria, comprised three taxa of gamma-proteobacteria (R-type [PASS], T-type [PABS], and U-type [PAUS]) and a rickettsia (S-type [PAR]). Complementary analysis of aphids from natural populations of four aphid species (A. pisum [n = 74], Amphorophora rubi [n = 109], Aphis sarothamni [n = 42], and Microlophium carnosum [n = 101]) from a single geographical location revealed Buchnera and up to three taxa of accessory bacteria, but no other bacterial taxa, in each aphid. The prevalence of accessory bacterial taxa varied significantly among aphid species but not with the sampling month (between June and August 2000). These results indicate that the accessory bacterial taxa are distributed across multiple aphid species, although with variable prevalence, and that laboratory culture does not generally result in a shift in the bacterial community in aphids. Both the transmission patterns of the accessory bacteria between individual aphids and their impact on aphid fitness are suggested to influence the prevalence of accessory bacterial taxa in natural aphid populations.

Animals↗
PubMed2003

Elucidation of the transmission patterns of an insect-borne bacterium.

Quantitative data on modes of transmission are a crucial element in understanding the ecology of microorganisms associated with animals. We investigated the transmission patterns of a gamma-proteobacterium informally known as pea aphid Bemisia-like symbiont (PABS), also known as T-type, which is widely but not universally distributed in natural populations of the pea aphid, Acyrthosiphon pisum. The vertical transmission of PABS to asexual and sexual morphs and sexually produced eggs was demonstrated by a diagnostic PCR-based assay, and the maximum estimated failure rate was 2%. Aphids naturally lacking PABS acquired PABS bacteria administered via the diet, and the infection persisted by vertical transmission for at least three aphid generations. PABS was also detected in two of five aphid honeydew samples tested and in all five siphuncular fluid samples tested but in none of 15 samples of salivary secretions from PABS-positive aphids. However, PABS-negative aphids did not acquire PABS when they were cocultured with PABS-positive aphids; the maximal estimated level of horizontal transmission was 18%. A deterministic model indicated that the force of infection by a horizontal transmission rate of 3% is sufficient to maintain a previously described estimate of the prevalence of PABS-positive aphids (37%), if the vertical transmission rate is 98%. We concluded that PABS infections in A. pisum can be maintained by high vertical transmission rates and occasional horizontal transmission, possibly via the oral route, in the absence of selection either for or against aphids bearing this bacterium.

Animals↗
PubMed2003

Metabolite comparisons and the identity of nutrients translocated from symbiotic algae to an animal host.

Dinoflagellate algae of the genus Symbiodinium in symbiosis with marine animals release much of their photosynthetic carbon to the animal host. The compounds translocated to the host ('mobile compounds') were investigated by metabolite comparison as follows: a substrate was identified as a candidate mobile compound when comparable profiles of metabolites were generated from host metabolism of this substrate (supplied exogenously) and the endogenous mobile compounds. When the sea anemone Anemonia viridis was incubated with NaH14CO2 under photosynthesizing conditions, most of the radioactivity in the animal tissue was recovered from the low-molecular-mass fraction and distributed in the ratio 1:2:1 between the neutral, acidic and basic sub-fractions. Prominent 14C-labelled compounds included glucose, malate and glucose-6-phosphate. When the symbiosis was incubated with 14C-labelled glucose plus succinate or fumarate (but none of eight other substrate combinations tested), the 14C-labelled metabolites closely matched those obtained with NaH14CO2. These data suggest that glucose and succinate/fumarate (or metabolically allied compounds) may be important photosynthetic compounds transferred from the Symbiodinium cells to the tissues of A. viridis. Metabolite comparisons can be applied to study nutritional interactions in symbioses involving photosynthetic algae and, with appropriate modification, other associations between microorganisms and plants or animals.

Animals↗
PubMed2002

Amino acid composition and nutritional quality of potato leaf phloem sap for aphids.

To define plant 'nutritional quality' for aphids, the causal basis of the variation in aphid performance between host plants of different developmental ages was explored using the aphids Myzus persicae and Macrosiphum euphorbiae on potato plants (Solanum tuberosum). Both aphid species performed better on developmentally young ('pre-tuber-filling') plants than on mature ('tuber-filling') plants. Aphid performance did not vary with leaf phloem sucrose:amino acid ratio but could be related to changes in the amino acid composition of the phloem, which included a developmental shift from high glutamine levels in pre-tuber-filling plants to low glutamine levels in tuber-filling plants. Aphid performance on chemically defined 'young' and 'old' diets, with amino acid composition corresponding to that of phloem amino acid composition in pre-tuber-filling and tuber-filling plants, respectively, confirmed that phloem amino acid composition contributed to low aphid performance on tuber-filling plants. The relatively poor performance on 'old' diets could be accounted for, at least in part, by depressed feeding rates. These data suggest that amino acid composition of the phloem is one factor shaping the nutritional quality of plants for aphids.

Amino Acids↗
PubMed2001

The impact of host plant on the abundance and function of symbiotic bacteria in an aphid.

The black-bean aphid Aphis fabae bears populations of coccoid symbiotic bacteria Buchnera spp. at 2.0-3.2 x 10(7)cells mg(-1)aphid mass and rod-shaped secondary symbionts of uncertain taxonomic affiliation at 0.1-0.6 x 10(7)cells mg(-1)aphid mass. Buchnera provides essential amino acids, supplementing the poor supply in the aphid diet of plant phloem sap. Comparison of the performance of A. fabae containing and experimentally deprived of their bacteria showed that the bacteria caused increased larval mass of aphids reared on Chenopodium album and Papaver dubium plants, but not when reared on Lamium purpureum. In the aphids reared on L. purpureum, the density of the bacteria, especially the secondary symbionts, was significantly elevated, and bacterial-mediated production of the essential amino acid threonine was reduced, even though the essential amino acid content of phloem exudates from L. purpureum had a low threonine content. It is proposed that the shortfall in threonine, possibly compounded by the high density of secondary symbionts, may contribute to the poor performance of the aphids on L. purpureum. This study offers the first evidence to suggest plant-mediated interference with the nutritional function of symbiotic bacteria in any phytophagous insect.

Amino Acids↗
PubMed2001

Quantifying nutrient production by the microbial symbionts in an aphid.

The symbiotic bacteria Buchnera sp. provide aphids with essential amino acids, nutrients in short supply in the aphid diet of plant phloem sap. The contribution of Buchnera-derived amino acids to net protein growth of the aphid Aphis fabae was quantified from the protein growth of aphids reared on chemically defined diets lacking individual amino acids. The amino acid production rates varied among the nine essential amino acids over the range 8-156 pmol microg(-1)protein day(-1) (for tryptophan and leucine, respectively), equivalent to 0.02-0.33 fmol Buchnera(-1)day(-1). In a complementary metabolic analysis, the aphids incorporated radioactivity from dietary [(14)C]glutamic acid into the essential amino acids isoleucine, lysine and threonine. Incorporation into isoleucine was significantly elevated by the omission of dietary isoleucine, indicating that dietary supply may affect the biosynthetic rates of certain amino acids by Buchnera. Aphids experimentally deprived of Buchnera did not synthesize essential amino acids from dietary glutamic acid. The mortality of aposymbionts was high over 7 days on the phenylalanine-free diet, and their assimilation of dietary leucine was depressed on the complete diet, suggesting that both the absence of bacteria-derived amino acids and the low rates of assimilation of certain dietary amino acids may contribute to the poor growth of these insects.

Amino Acids, Essential↗
PubMed1999

Low genetic diversity among pea aphid (Acyrthosiphon pisum) biotypes of different plant affiliation.

Genetic diversity in the pea aphid Acyrthosiphon pisum was investigated by a restriction fragment length polymorphism (RFLP) analysis of three maternally inherited genomes (mitochondrial DNA and plasmids of the symbiotic bacteria Buchnera). Twenty-nine parthenogenetic clones of three A. pisum biotypes, defined by their capacity to use the legume crops pea, alfalfa and red clover, respectively, were analysed, and a total of 67 restriction sites was scored. No restriction site variation in the mitochondrial genome was obtained, but length variation at two regions (the A + T-rich region and ND3-ND5 region) was noted. One aphid clone bore a variant HindIII restriction site in the Buchnera leucine plasmid (pAPEleu), and two clones were heteroplasmic for a 0.76-kb deletion in the Buchnera tryptophan plasmid (pAPEtrp). Based on arthropod nucleotide substitution rates, it is proposed that the crop-feeding biotypes of A. pisum may have diversified within the last 100 000 years and possibly much more recently, since the advent of agriculture.

Animals↗
PubMed1998

Nutritional interactions in insect-microbial symbioses: aphids and their symbiotic bacteria Buchnera.

Most aphids possess intracellular bacteria of the genus Buchnera. The bacteria are transmitted vertically via the aphid ovary, and the association is obligate for both partners: Bacteria-free aphids grow poorly and produce few or no offspring, and Buchnera are both unknown apart from aphids and apparently unculturable. The symbiosis has a nutritional basis. Specifically, bacterial provisioning of essential amino acids has been demonstrated. Nitrogen recycling, however, is not quantitatively important to the nutrition of aphid species studied, and there is strong evidence against bacterial involvement in the lipid and sterol nutrition of aphids. Buchnera have been implicated in various non-nutritional functions. Of these, just one has strong experimental support: promotion of aphid transmission of circulative viruses. It is argued that strong parallels may exist between the nutritional interactions (including the underlying mechanisms) in the aphid-Buchnera association and other insect symbioses with intracellular microorganisms.

Journal Article↗
PubMed1997

Partitioning of Symbiotic Bacteria between Generations of an Insect: a Quantitative Study of a Buchnera sp. in the Pea Aphid (Acyrthosiphon pisum) Reared at Different Temperatures.

The population of symbiotic Buchnera bacteria in parthenogenetic females of the pea aphid Acyrthosiphon pisum was determined by quantitative hybridization of a DNA probe (groESL) to aphid homogenates. The aphids bore 1 x 10(sup7) to 2 x 10(sup7) bacterial cells per mg (fresh weight). In teneral aphids (i.e., aphids that had moulted to adulthood but that had not initiated reproduction), >75% of the bacteria were in the embryos, and the density of bacteria in the embryos was consistently greater than that in the maternal tissues. The bacterial density in teneral aphids increased from 1.3 x 10(sup7) to 2.0 x 10(sup7) cells mg (fresh weight) of aphids(sup-1) with temperature between 15 and 25(deg)C. This variation could be attributed to a temperature-dependent increase in both the density of bacteria in the embryos and embryo content of the aphids.

Journal Article↗
PubMed1993

Effects of the fungus Aspergillus penicillioides on the house dust mite Dermatophagoides pteronyssinus: an experimental re-evaluation.

In this report the widely-held view that house dust mites benefit from fungal contamination of the dietary substratum is re-examined. The performance of Dermatophagoides pteronyssinus (Acari: Pyroglyphidae) is documented over two successive generations in the presence or absence of the xerophilic fungus Aspergillus penicillioides (Hyphomycetales: Moniliaceae). This fungus reduced survival, development rate, adult length and fecundity of D. pteronyssinus. Detrimental effects of A. penicillioides were proportional to the fungal density. Despite the antagonistic effects of A. penicillioides, a requirement for the fungus was indicated by the poor performance of fungus-free mites in the second generation; sustained culture of D. pteronyssinus in the absence of fungi is probably not possible. It is suggested that fungi may alter the particulate nature of the substratum to the detriment of house dust mites, but also provide micronutrients deficient in the diet.

Animals↗
PubMed1992

How relevant are house dust mite-fungal interactions in laboratory culture to the natural dust system?

Both house dust and house dust mites Dermatophagoides pteronyssinus contained a wider range of fungi than laboratory mite cultures. In total, nine species of fungi were isolated from D. pteronyssinus in house dust, and these included three xerophilic species (Eurotium amstelodami, Aspergillus penicillioides and Wallemia sebi) commonly found in laboratory cultures of D. pteronyssinus. It is concluded that mites do interact with a similar range of fungi in natural dust and in laboratory culture, but that the diversity of fungal species in the laboratory is reduced and the density of individual fungal species in culture exceeds that of house dust. In a second experiment, dust samples were incubated at room temperature with 75% relative humidity. The diversity of fungi invariably declined from up to 13 genera to the few species recorded in laboratory culture. This suggests that the dominance of xerophilic fungi in laboratory mite rearings is mediated primarily by low relative humidity, and the exclusion of air-borne spores.

Air Microbiology↗