Predation of bacteria by the protozoa Tetrahymena pyriformis in toluene-degrading cultures

The influence of the toluene concentration on predation of toluene-degrading bacteria by the protozoa Tetrahymena pyriformis was investigated in suspended batch cultures continuously aerated with toluene-contaminated air. At gas phase concentrations of 0.035 to 0.74 g m–3, toluene did not significantly affected protozoan activity and the final bacteria concentration was reduced by growing protozoa by 98 to 99.9% compared to protozoa-free controls. As the toluene concentration was increased to 1.16–1.33 g m–3, the reduction of the bacteria cell concentration was 80%. At 3.35 g toluene m–3, growth of T. pyriformis was completely inhibited. Overall, the results presented herein demonstrate that protozoa grazing on bacteria play a major role in controlling bacterial cell concentration, but that the toxicity of the treated pollutants to the protozoa is an important factor that needs to be taken into account in biological treatment processes.     

Larval mortality of Culex tarsalis and Aedes aegypti when reared with different concentrations of Tetrahymena pyriformis

The paucity of information on infection and pathogenicity of mosquito larvae by ciliates under experimental conditions prompted the evaluation of the relationship between a clone of Tetrahymena pyriformis and two species of mosquitoes, Aedes aegypti and Culex tarsalis. A significant difference in mortality was observed between the two mosquito species when they were exposed to identical concentrations of ciliates. In rearing trays containing high concentrations of T. pyriformis, 76.3% of the C. tarsalis larvae died. Under similar conditions, death occurred in less than 3.0% of the A. aegypti larvae. Three figures illustrate daily results of these studies. Three criteria for facultative parasites are listed, and one, route of entry into the host, is discussed in detail.     

Toxicity of Bacillus thuringiensis toward Aedes aegypti larvae.

Among six strains of Bacillus thuringiensis and five other species of Bacillus, only two strains of B. thuringiensis, strains HD-1 and BA-068, were toxic to Aedes aegypti larvae within 24 hr. The LC₅₀s were 5.6 × 10⁴ and 2.4 × 10⁵ spores/ml for strains HD-1 and BA-068, respectively. The toxic factor(s) was heat sensitive and γ ray resistant and preliminary evidences indicated that it was associated with the crystalline body of B. thuringiensis.     

Susceptibility and alterations by diflubenzuron in larvae of Aedes aegypti

Benzoylphenyl ureas inhibit chitin synthesis and interfere with the molting process in arthropods. In this study, the effect of diflubenzuron on third‐instar larvae of Aedes aegypti was evaluated. The susceptibility to the product was determined, and the alterations generated were shown through light and electron microscopy. LC₅₀ and LC₉₀ were 0.23 and 0.47 ppm, respectively. The main alterations observed were the incapacity to complete the molt, a reduction of mobility, the fragmentation of the old cuticle, a division of the body segments that was not evident, and the deformation of the caudal structures. Images of the ultrastructure are included, where breaking zones in the cuticle were observed, separation of the cuticle, the epidermis and the muscles, and these latter with a disorganized arrangement. In low concentrations, from 0.15 ppm, diflubenzuron causes alterations in the behavior and morphology of Ae. aegypti.     

Major hemolymph protein of Aedes aegypti larvae

Aedes aegypti larval hemolymph proteins were analyzed, and the major protein was characterized. The major protein, designated P1, is hexameric and is composed of subunits with molecular weights estimated to be 83,000. P1 is dissociated into its subunits when the pH is elevated from 7 to 9. This protein accumulates during the last larval instar and is not detected in adult mosquitoes. These characteristics, together with the high content of aromatic amino acids, include P1 in the arylphorin group of the insect storage proteins. Arch. Insect Biochem. Physiol. 34:191–201, 1997. © 1997 Wiley-Liss, Inc.     

Digestion of Bacillus thuringiensis var. israelensis spores by larvae of Aedes aegypti.

The larvicidal activity of Bacillus thuringiensis var. israelensis against mosquitoes and the blackfly is included in parasporal crystalline bodies which are produced during sporulation. Following ingestion, the crystals are solubilized in the larval midgut and induce death within a short time; the spores germinate in the dead larvae and complete a growth cycle. The fate of the spores in surviving live larvae was elucidated by using a nonlarvicidal B. thuringiensis var. israelensis mutant. When introduced as the only food source, spores of this mutant support development to the adult stage of newly hatched Aedes aegypti larvae at a rate directly related to spore concentration. The conclusion that spores of B. thuringiensis var. israelensis are digested in the larval gut was substantiated by following the incorporation of [35S]methionine-labeled spores into larval tissues.     

Properties of a ribonuclease from Aedes aegypti larvae

1. The properties of a soluble ribonuclease from Aedes aegypti larvae have been compared with ribonuclease activity in adult female tissue. 2. In larval extracts ribonuclease activity was maximal at 40-45 degrees C whereas activity in tissue from adult females was highest at 50 degrees C. 3. Ribonuclease activity that was recovered in a 20-60% ammonium sulfate precipitate was further purified by batch elution from DEAE-Sephacel and from carboxymethylcellulose. 4. Ribonuclease activity in the partially purified fraction was sensitive to EDTA, stimulated by magnesium, had a pH optimum at 9.0 and a Mr of 45,000. 5. Agarose gels containing yeast RNA substrate were used to monitor partial purification of the larval ribonuclease.     

Persistence and pathogenicity of a native isolate of Leptolegnia chapmanii against Aedes aegypti larvae in different anthropic environments

The oomycete Leptolegnia chapmanii has been identified as a potential control agent of the primary vector of dengue, Aedes aegypti. In our assays, the persistence and pathogenicity of a native isolate of L. chapmanii decreased over time regardless of location. However, the mortality of Ae. aegypti larvae was significantly lower (p < 0.05) in containers located outside without sun protection (89% at first week and 9% at sixth week) compared with the containers located indoors (97% at first week and 42% at sixth week) and outside with shade (89% at first week and 29% at sixth week) possibly because of exposure to sun radiation.     

Proteome of Aedes aegypti larvae in response to infection by the intracellular parasite Vavraia culicis

We report on the modification of the Aedes aegypti larval proteome following infection by the microsporidian parasite Vavraia culicis. Mosquito larvae were sampled at 5 and 15 days of age to compare the effects of infection when the parasite was in two different developmental stages. Modifications of the host proteome due to the stress of infection were distinguished from those of a more general nature by treatments involving hypoxia. We found that the major reaction to stress was the suppression of particular protein spots. Older (15 days) larvae reacted more strongly to infection by V. culicis (46% of the total number of spots affected; 17% for 5 days larvae), while the strongest reaction of younger (5 days) larvae was to hypoxia for pH range 5-8 and to combined effects of infection and hypoxia for pH range 3-6. MALDI-TOF results indicate that proteins induced or suppressed by infection are involved directly or indirectly in defense against microorganisms. Finally, our MALDI-TOF results suggest that A. aegypti larvae try to control or clear V. culicis infection and also that V. culicis probably impairs the immune defense of this host via arginases-NOS competition.     

Dye-induced changes in the developmental physiology of Aedes aegypti larvae

Exposure to methylene blue and neutral red affected length of development, rate of pupation, and larval mortality in populations of Aedes aegypti (L.). Female pupal weights generally were adversely affected, while male pupal weights were not. Retardation of growth was not caused by rejection of dyed food under the conditions of our experiments. Methylene blue, neutral red, and nile blue A were most severe in their action on longer exposures and exposures to earlier instars.The importance of recognizing the physiological and behavioral changes in organisms caused by perfunctory use of dyes is discussed.

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Zusammenfassung Bei Larven von Aedes aegypti, die Methylenblau oder Neutralrot ausgesetzt wurden, ließ sich eine deutliche Verzögerung des Wachstums nachweisen. Der Verpuppungsbeginn (Larven-Puppen-Häutung) wurde von beiden Farben in Abhängigkeit von der ansteigenden Konzentration verzögert. Obwohl zur Erzeugung der Reaktion mit Neutralrot höhere Konzentrationen erforderlich waren, war die Genauigkeit der Farbwirkung größer. Die geprüften Konzentrationen von Methylenblau reichten von 0,5 bis 4,5 ppm; die für Neutralrot von 3 bis 9 ppm.In der Absicht, die Wirkungen der beiden Farben zu messen, wurden andere Parameter quantitativ geprüft. Diese umfaßten die Mortalität, den Weibchen-Prozentsatz und die durchschnittlichen Puppengewichte der Männchen. Die Sterberaten waren hoch und äußerst variabel. Es ließen sich auch keine Unterschiede im Geschlechterverhältnis der Populationen finden, die als Larven in Methylenblau oder Neutralrot aufgezogen worden waren. Neutralrot und Methylenblau schienen auch die durchschnittlichen Puppengewichte der Männchen nicht zu beeinflussen, jedoch erzeugten sie deutliche Wirkungen bei den durchschnittlichen Puppengewichten der Weibchen. Es konnten keine signifikanten Unterschiede in den Nahrungsmengen festgestellt werden, die von gefärbten oder ungefärbten Larven oder von Larven in ansteigenden Farbkonzentrationen aufgenommen wurden. Die jüngeren Larvenstadien wurden stärker beeinflußt und längerer Aufenthalt in der Farbe ergab stärkere Verzögerung der Wachstumsrate.Folgende Aspekte der Vital-Farbstoffe werden diskutiert: 1. ihre toxischen Wirkungen, 2. Beziehungen zwischen Genauigkeit und Aussagewert der experimentellen Ergebnisse, und 3. die Notwendigkeit vollständigerer Kenntnis der Farbstoffe vor ihrer Anwendung auf lebende Systeme.

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Contribution No: 1420 from the Department of Entomology, University of Massachusetts, Amherst, Mass. This research was supported by Hatch Project No. 253 Revised.     

Sublethal metal stress response of larvae of Aedes aegypti

Aedes aegypti (Diptera: Culicidae) has adapted to urban environments. The urbanization process provides suitable habitats for this disease vector, subsequently increasing the probability of the transmission of pathogens in high‐density environments. Urban environments provide metal‐stressed larval habitats. However, little is known about the physiological cost of metal stress or how this might affect the performance of this mosquito species. The present study aims to characterize the sublethal physiological consequences of metal stress in Aedes aegypti. Various parameters of mosquito physiology under larval metal stress are assessed, including larval metallothionein expression and the effects of larval metal stress on adult performance and their progeny. The results show that environmentally relevant larval metal stress compromises larval and adult development and performance, and also results in larval metal tolerance along with an increase in lipid consumption. These performance costs are coupled with a dramatic increase in metallothionein expression in the midgut. Metal stress results in a lowered adult body mass and neutral storage lipids at emergence, starvation tolerance, fecundity and starvation tolerance of offspring compared with non‐metal‐stressed individuals. Ironically, larval metal stress results in increased adult longevity. Taken together, these findings indicate that even low levels of environmentally relevant larval metal stress have considerable physiological consequences for this important disease vector.