Ecology of the mycophagous nematodeAphelenchus avenae in wheat-field and pine-forest soils

Summary The soils of a wheat-field and pine-forest had different mycofloras and supported different populations ofAphelenchus avenae, a mycophagous nematode. The abundance ofA. avenae was correlated with the composition of the mycoflora in these soils; the greater abundance ofA. avenae in wheat-field soil being associated with a more diverse mycoflora.     

Coiling is Not Essential to Anhydrobiosis by Aphelenchus avenae on Agar Amended with Sucrose

The roles of preconditioning and coiling upon entrance into anhydrobiosis by Aphelenchus avenae were tested via video-assisted analysis at 25²°C. Fourth-stage juveniles or young adults of A. avenae were individually placed on 5% agar containing 0.8 M sucrose. Nematodes became quiescent within 3 hr, then gradually resumed a low level of activity and assumed a coiled posture. High desiccation survival rate was recorded when nematodes were incubated on agar for more than 6 hr; the survival rates were 0%, 3%, 73%, and 92% for 0, 2, 6, and 12 hr on agar, respectively. All nematodes placed on agar for 24 hr or more revived after rehydration following desiccation. Once nematodes were on agar for a sufficient time, no difference in desiccation survival was observed between nematodes taking a coiled posture and those uncoiled artificially. Based on these results, exposure to osmotic stress for 6 hr can prepare A. aveae physiologically for anhydrobiosis, but coiling does not appear to be a physiological requirement for desiccation in survival.     

Inhibition of Acetylcholinesterases from Aphelenchus avenae by Carbofuran and Fenamiphos

Exposure to carbofuran and fenamiphos for 72 hours reduced the numbers of active Aphelenchus avenae in aqueous suspension by > 75%. When nematicides were removed, many A. avenae exposed to carbofuran resumed normal movement but A. avenae treated with fenamiphos did not recover. Acetylcholinesterase (AChE) activity was suppressed by > 95% in nematodes treated with carbofuran or fenamiphos. However, 48 hours after treated nematodes had been placed in water, AChE activity in carbofuran treated populations was 98% of the levels in control nematodes. Nematodes that had been treated with fenamiphos showed only slight AChE recovery. The antidotes, atropine sulfate and 2-PAM, were largely ineffective in counteracting the toxic effects of the nematicides.     

Feeding behavior and transmission of barley yellow dwarf virus by Sitobion avenae on oats

Feeding behavior of Sitobion avenae F. (Homoptera: Aphididae) on oats (Avena sativa cv. Clintland 64) was electronically monitored, and waveforms corresponding to salivation, ingestion, and sieve element penetration described.During 90 min plant access, aphids ingested from phloem for 0–43 min (mean: 8.1 min) and non-phloem for 0–60 min (mean: 19 min). Only 65% of the aphids tested made phloem contact within 90 min, contacting phloem after 18–85 min (mean: 32 min). No significant difference was observed in the feeding behavior of aphids carrying barley yellow dwarf virus (BYDV) from that of non-viruliferous aphids.Penetration of a sieve element was a prerequisite for BYDV transmission but did not insure transmission. Penetration of one sieve element resulted in a 65% chance of transmission independent of the duration of phloem contact. The chance of transmission increased with increasing number of sieve element penetrations.Inoculation of oat seedlings with single, viruliferous aphids for 90 min is estimated to cause 54% of the plants to be infected. Also, it is estimated that no transmission can occur with plant access periods shorter than 17 min.

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Zusammenfassung Das Probeverhalten von Sitobion avenae F. (Homoptera: Aphididae) auf Hafer (Sorte Clintland 64) wurden elektronisch verfolgt. Drei Wellenformen (S=Speichelfluss, X-das Eindringen in ein Siebelement, I=Nahrungsaufname) wurden registriert. In histologischen Untersuchungen wurden diese Wellenformen mit der Position der Stechborsten korreliert. Wenn X-Wellen oder eine X-I-Folge registriert wurden, war die Stechborste immer im Phloëm; bei S-Wellen oder bei einer S-I-Folge, war die Stechborste nie im Phloëm.Die Blattläuse wurden auf gesunden, fünf Tage alten Pflanzen während 90 Min beobachtet. 65% der Blattläuse erreichten nach 18–85 Min (Durchschnitt 32 Min) das Phloëm. Innerhalb 90 Min nahmen die Blattläuse während 0–43 Min (Durchschnitt 8.1 Min) Saft aus dem Phloem und während 0–60 Min (Durchschnitt 19 Min) Saft aus dem Mesophyll auf. Keine signifikanten Unterschiede im Probeverhalten wurden bei Blattläusen mit und ohne Barley Yellow Dwarf Virus (BYDV) festgestellt.Blattläuse, die innerhalb 24 Stunden BYDV übermitteln können, wurden für Übertragungsversuche verwendet. Das Probeverhalten dieser Blattläuser wurde manipuliert, und die Leistungsfähigkeit der Übertragung mit einer immunologischen Technik (ELISA) untersucht. Um BYDV zu übertragen mussten die Blattläuse mit einem Siebelement in Kontakt kommen. Nach der Stechborstenpenetration in ein Siebelement wurden 65% der Pflanzen mit BYDV infiziert. Der Prozentsatz infizierter Pflanzen und der Virusiter in den infizierten Pflanzen waren mit der Dauer der Siebelement-penetration (Anzahl von X-Wellen) nicht proportional. Wenn die Blattläuse mit zwei oder drei Siebelementen in Kontakt kamen, wurde der Prozentsatz infizierter Pflanzen signifikant erhöht, während der Virustiter nich verändert wurde. Infektionsprozente niedriger als 100% nach Siebelement-penetration sind möglicherweise das Resultat von Unterschieden in den Siebelementen.Es wird geschätzt dass 50% Infektion eintritt, wenn Pflanzen während ungefähr 83 Min von einer einzelnen infizierten Blattlaus besogen werden. Keine Übertragung von BYDV kann eintreten, wenn die Probezeit weniger als 17 Minuten beträgt.

     

Antagonistic actinomycetes from Moroccan soil to control the grapevine gray mold

One hundred and forty-two different actinomycete strains were isolated from rhizosphere soil of Vitis vinifera L. sampled from four Moroccan areas. To evaluate the antifungal effect of the different collected actinomycete isolates, five fungi known to be phytopathogens (Pythium ultimum, Fusarium oxyysporum f. sp. albedinis, Sclerotium rolfsii, Verticillium dahliae and Botrytis cinerea) were used. Results showed that 24 isolates had an in vitro inhibitory effect toward at least 4 of the indicator fungi, but only 9 inhibited all these phytopathogens. These nine isolates were subsequently evaluated individually using in vitro grapevine plantlets for their ability to protect against plant gray mold. We demonstrate here that pre-inoculation of plantlets with these isolates allow them to withstand Botrytis cinerea. Six of these strains were shown to belong to the genus Streptomyces and three to the genus Micromonospora. These findings indicate the potential of developing effective actinomycetes from Moroccan habitats for the biological control of Botrytis cinerea. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Airborne dust,bacteria, actinomycetes and fungi at a flourmill

A study was carried out on suspended dust, bacterial and fungal aerosols in a four-storey flourmill building located in Giza, Egypt. Airborne microorganisms were quantitatively isolated using liquid impinger and gravimetric samplers during the period from March 2004 to February 2005. Suspended dust varied from 1.96 to 16.3 mg m⁻³ and 0.69 to 1.8 mg m⁻³ in the indoor and outdoor environments, respectively. Suspended dust was significantly greater (P < 0.05) at bran package, double roller, purifiers and flour storage units in comparison to the outdoor reference site. The dust levels exceed the occupational exposure limit (OEL) of 0.5 mg m⁻³ for flour dust. Airborne microbial counts were found at median values, between sampling locations, ranged from 0 to >10⁴ CFU m⁻³. Gram-negative bacteria were found in small numbers (0–10² CFU m⁻³). The highest concentration of actinomycetes (>10³ CFU m⁻³) was detected in the storage unit. Airborne fungal counts were found at the median values, between sampling locations, varied from 10³ to 10⁴ CFU m⁻³. The counts of airborne bacteria and fungi were significantly greater (P < 0.05) at the purifiers and double roller mill units in comparison to the outdoor reference site using the liquid impinger sampler. Microbial levels associated with bulk deposited dust averaged between 10⁵ and 10⁶ CFU g⁻¹. Alcaligenes (5.4%) Pseudomonas (3.87%) and Enterobacter (3.1%) were the predominant Gram-negative species while Bacillus (29.4%) and Micrococci (13.9%) were the major components of Gram-positive bacteria. Aspergillus and Penicillium were the predominant fungal types indoor whereas Cladosporium (35.2%) and Aspergillus species (22.2%) were the predominant fungal types outdoor. A number of allergenic and toxigenic bioaerosols were found in the flourmill workplace.     

Cuticle Formation in Hemicycliophora arenaria,Aphelenchus avenae and HirschmannIella gracilis

The onset of molting in all stages of Hemicycliophora arenaria was preceded by the appearance of numerous, discrete globular structures which were termed "molting bodies" because they were present in the hypodermis only during the production of the new cuticle. In all parasitic stages the molt commenced with the separation of the cuticle from the hypodermis from which the new sheath and cuticle were differentiated. Following completion of the new sheath and cuticle most of the old outer covering was apparently absorbed before ecdysis. Electronmicrographs of body wall cross sections in molting L4 male specimens revealed the final molt to be a double molt in which an additional sixth cuticle was produced. Since both a new sheath and cuticle were produced during the molt of each stage, the sheath must be considered as an integral part of the cuticle and not as a residual cuticle or the result of an incomplete additional molt. Molting in Aphelenchus avenae and Hirschmanniella gracilis was less complex and "molting bodies" were not observed. After cuticle separation the hypodermis gave rise to a new trilaminate zone, the future cortex, and (later) the matrix and striated basal layers.     

Characterization of selected actinomycetes degrading polyaromatic hydrocarbons in liquid culture and spiked soil

Summary Five strains of the Rhodococcus and Gordonia genera were evaluated for their potential use in bioremediation of polycyclic aromatic hydrocarbons (PAH) with or without another substrate (co-substrate). Their ability to produce biosurfactants or to degrade phenanthrene when growing on glucose, hexadecane and rapeseed oil was tested in liquid medium at 30 °C. All strains showed biosurfactant activity. The highest reduction in surface tension was recorded in whole cultures of Rhodococcus sp. DSM 44126 (23.1%) and R. erythropolis DSM 1069 (21.1%) grown on hexadecane and Gordonia sp. APB (20.4%) and R. erythropolis TA57 (18.2%) grown on rapeseed oil. Cultures of Gordonia sp. APB and G. rubripertincta formed emulsions when grown on rapeseed oil. After 14 days of incubation, Rhodococcus sp. DSM 44126 degraded phenanthrene (initial concentration 100 μg ml⁻¹) as sole carbon source (79.4%) and in the presence of hexadecane (80.6%), rapeseed oil (96.8%) and glucose (below the limit of detection). The other strains degraded less than 20%, and then with a co-substrate only. Rhodococcus sp. DSM 44126 was selected and its performance evaluated in soil spiked with a mixture of PAH (200 mg kg⁻¹). The effect of the addition of 0, 0.1 and 1% rapeseed oil as co-substrate was also tested. Inoculation enhanced the degradation of phenanthrene (55.7% and 95.2% with 0.1% oil and without oil respectively) and of anthracene (29.2% with 0.1% oil). Approximately 96% of anthracene and 62% of benzo(a)pyrene disappeared from the soil (inoculated and control) after 14 days and anthraquinone was detected as a metabolite. Rhodococcus sp. DSM 44126 was identified as Rhodococcus wratislaviensis by 16S rRNA sequencing and was able to degrade anthracene as sole carbon source in liquid culture.     

Sensitivity of Acetylcholinesterases from Aphelenchus avenae to Organophosphorous and Carbamate Pesticides

The sensitivities of acetylcholinesterases (ACHE) from the nematode Aphelenchus avenae and the house fly Musca domestica to various pesticides were compared using a colorimetric assay. ACHE from A. avenae were generally less sensitive than ACHE from M. domestica to inhibition by organophosphorous and carbamate pesticides. Carbamates were somewhat more inhibiting than organophosphorous pesticides to nematode ACHE. In vivo tests with concentrations of various pesticides up to 500 ppm in sand caused less than 100% mortality of nematodes.     

Mobility of apterous grain aphids Sitobion avenae within wheat fields

In a field study of the grain aphid Sitobion avenae (F.) almost all apterous aphids were found to move from the natal ear before reproducing, and alatiform nymphs usually moved between ears during their development. Apterae were very mobile, with over 50% staying on one ear for less than 24 h. Movements did not appear to be a response to climatic conditions, predation or colony density.

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Résumé Lors de l'étude de Sitobion avenae dans la nature, on a constaté que presque tous les individus aptères avaient quitté leur épi d'origine avant la reproduction, et que les larves de type ailé s'étaient déplacées généralement entre les épis pendant leur développement. Les aptères étaient très mobiles, la majorité restant moins de 24 h sur le même épi. Ces déplacements n'ont pas semblé dépendre des conditions climatiques, de la prédation ou de la densité des colonies.

     

Production of sexual morphs by the monoecious cereal aphid Sitobion avenae

The effects of photoperiod and temperature on the production of sexual forms by two clones of Sitobion avenae, the grain aphid, were examined. One clone did not produce sexual forms, whereas the other did under conditions of short light (<14 h) and low temperature (15°C). Temperature and photoperiod interacted to some extent both in the production of oviparae and of males. Even when the sexual forms were produced there was always a proportion of virginoparae.

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Production de sexués par Sitobion avenae, puceron monoécique

Résumé Un clone de Sitobion avenae a produit sans difficulté jusqu'à 7 générations de sexués en réaction à des photopériodes courtes (<14 h) et à de faibles températures (15°C) tandis qu'un autre clone n'en produit aucune.La proportion de sexués différait suivant les parents. La production de mâles avait une nette tendance à apparaître lors des reproduction les plus tardives. Dans les générations tardives avec jours courts, les vivipares (virginopares et gynopares) avaient eu tendance à être produits à la fin de la période de reproduction.Les gynopares (c'est à dire les parents d'ovipares) de S. avenae étaient surtout aptères, mais comprenaient beaucoup plus d'ailés que les virginopares obtenus dans les mêmes conditions. Un vivipare était induit comme gynopare (ou ses embryons étaient déterminés comme ovipares) avant la naissance, mais cette détermination pouvait apparemment encore être inversée en soumettant l'insecte à de longues photopériodes et à de hautes températures jusqu'à deux jours après la naissance.Aucun S. avenae ovipare n'a été produit jusqu'à la troisième génération. Aucun ovipare n'a été produit avec des photopériodes supérieures à 13 H 30 à 10°C, 13 H à 15°C et 8 H à 20°C. La proportion d'individus produisant des ovipares à 15°C a été plus faible qu'à 10°C pour toutes les photopériodes, et à cette dernière température beaucoup plus de vivipares étaient gynopares que virginopares.Les basses températures ont été vraisemblablement le facteur dominant de stimulation de la production de mâles de S. avenae, mais cependant il semble qu'un plus grand nombre de mâles a été produit aux températures et photopériodes intermédiaires qu'aux extrêmes.La capture de mâles ailés de S. avenae dans des pièges à succion a été généralement limitée à Octobre-soit à peu prés le moment prévu par les expériences de laboratoire. Des mâles de S. avenae sont aussi capturés fréquemment et été, ce qui peut être lié à des hivers précédents doux.

     

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

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

Studies on bacterioplankton and inhibitory strains of aquatic actinomycetes in Lake Bardawil,Egypt

Lake Bardawil represents a hyper-saline shallow (50–600 cm deep) water basin lying along the northern shore of Sinai. Surface water samples were collected monthly from this lake for physico-chemical and microbiological analysis. Seasonal variations in the averages of temperature, pH and transparency of the tested water samples were determined. Seasonal averages of the total bacterial counts at 22 and 37°C, biovolumes of cells and bacterial biomass of the lake’s water at different regions were also determined. In addition to the gram-negative pathogen, Edwardsiella tarda, faecal contamination indicators, total coliforms, faecal coliforms and faecal streptococci were counted in the lake’s water samples from various regions during different seasons. The number of actinomycetes per milliliter of the lake’s water samples were determined. Fifteen actinomycetes were isolated and screened for their antibacterial activity against E. coli and pathogenic E. tarda isolated from the same water samples. Five isolates of these actinomycetes showed antibacterial activities and nine of them were identified as Streptomyces. The most antibacterially active isolate was subjected to morphological, physiological and biochemical studies and identified as Streptomyces viridiviolaceus. The identified organism exhibited antimicrobial activities against the main indicator of water pollution, E. coli, pathogenic E. tarda, Corynebacterium michiganese B-33, Pseudomonas solanacearum B-3212 and Staphylococcus. No antimicrobial activities were recorded against Bacillus cereus, Micrococcus, Mycobacterium, Aspergillus niger, Candida albicans, C. tropicalis or Rhizopus nigricans.     

Reproductive stragegies of the alate and apterous morphs of the grain aphid, Sitobion avenae

Differences were observed in the total fecundity and patterns of reproduction of four morphs of Sitobion avenae (F.). The reproductive rates of apterous and alate exules declined gradually with time but decreased rapidly in gynoparae and oviparae. Under short daylengths apterous exules gave birth in three phases; the first consisting mainly of gynoparae, the second of males and the third of non-gynoparous viviparae. The strategic importance of the sequence of morphs and the reproductive behaviour of each is discussed in relation to the maximization of overwintering population size.

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Stratégies reproductives des types ailés et aptères de Sitobion avenae F.

Résumé Des différences ont été observées entre les fécondités totales et les modes de reproduction de quatre types de Sitobion avenae. Les taux de reproduction diminuent graduellement dans le temps chez les aptères et ailés virginopares et rapidement chez les gynopares et ovipares. En photophases courtes, les virginopares aptères donnent naissance à trois types: le premier composé essentiellement de gynopares, le second de mâles et le troisième de vivipares. L'importance stratégique de la succession des types et du comportement reproducteur de chacun est discutée en relation avec l'accroissement maximum de la taille de la population hivernante.

     

Cuticle Ultrastructure of Hemicycliophora arenaria,Aphelenchus avenae,Hirschmanniella gracilis and Hirschmanniella belli

Ultrathin sections of all parasitic stages of Hemicycliophora arenaria revealed two major divisions in the body covering. The outermost was a seven-layered sheath and the innermost a five-layered cuticle comprising three zones; an outer trilaminate cortex, a fibrillar matrix and a striated basal layer. The body covering of the nonparasitic males also exhibited two major divisions: the outer, a relatively thin four-layered sheath and the inner, a six-layered cuticle consisting of three zones; an outer trilaminate cortex, a two-layered matrix and a striated basal layer. The cuticles of all stages of Aphelenchus avenae were similar, consisting of five layers divisible into three zones; an outer trilaminate cortex, a fibrillar matrix and a striated basal layer. Hirschmanniella gracilis and H. belli cuticles were also similar in all stages examined, consisting of six layers divisible into three zones; an outer trilaminate cortex, a two-layered matrix and a striated basal layer.     

Isolation of endophytic actinomycetes from selected plants and their antifungal activity

The isolation of endophytic actinomycetes from surface-sterilized tissues of 36 plant species was made using humic acid–vitamin (HV) agar as a selection medium. Of the 330 isolates recovered, 212 were from roots, 97 from leaves and 21 isolates from stems with a prevalence of 3.9, 1.7 and 0.3%, respectively. Identification of endophytic actinomycetes was based on their morphology and the amino acid composition of the whole-cell extract. Most isolates were classified as Streptomyces sp. (n = 277); with the remainder belonging to Microbispora sp. (n = 14), Nocardia sp. (n = 8) and Micromonospora sp. (n = 4). Four isolates were unclassified and 23 were lost during subculture. The most prevalent group of isolates were the Streptomyces sp. occurring in 6.4% of the tissue samples of Zingiber officinale. Scanning electron microscopy investigation of this plant revealed that 7.5% of the root and 5% of the leaf samples contained endophytes. Three of the Streptomyces sp. isolates strongly inhibited Colletotrichum musae, five were very active against Fusarium oxysporum and two strongly inhibited growth of both test fungi.     

The spatial and temporal distribution of the grain aphid Sitobion avenae in winter wheat

The spatial and temporal distribution of the grain aphid Sitobion avenae F. (Homoptera: Aphididae) was studied within a field of winter wheat during the summer of 1996. Sampling was done using four nested grids comprising 133 locations. Analysis by Taylor's power law gave results typical for insect populations. Analysis by SADIE (Spatial Analysis by Distance Indices) showed spatial pattern due to edge effects and sampling scale, and positive but mild spatial association, although spatial patterns were ephemeral. Reasons for these findings and the implications for integrated crop management are discussed.     

Methods of hatching the eggs and rearing the fundatrices of the English grain aphid, Sitobion avenae

Several methods for hatching the eggs and rearing individuals of the first generation (fundatrices) of Sitobion avenae were investigated. The most successful methods were incubation of the eggs on grass seedlings at 2°C and rearing the fundatrices on grass seedlings (overall survival 66%) and incubation of the eggs in plastic boxes at 2°C and rearing the fundatrices on wheat seedlings (overall survival 62%).

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Résumé L'éclosion des oeufs de S. avenae peut être induite par le transfert à 10°C ou 12°C, après une incubation de 75–120 jours à 2°C. Le pourcentage le plus élevé d'éclosions a été obtenu quand les oeufs avaient incubé pendant 100 à 110 jours à 2°C (67% at 71.5% respectivement) dans des petites boîtes de plastique, ou pendant 100 jours à 2°C sur des pousses de graminées (73.5%). Si les oeufs sont pondus sur blé, la plante ne peut pas tolérer la période d'incubation, mais cet obstacle peut être surmonté en obligeant les ovipares à pondre leurs oeufs sur de pousses de graminées, comme Poa annua, hôte convenable pour les fondatrices. Les ovipares peuvent aussi pondre sans difficultés sur autre chose que des végétaux, et des récipients peuvent ètre mis à incuber sans contenir du matériel végétal.

     

An investigation of the differential performance of clones of the aphid Sitobion avenae on two host species

Individuals of the grain aphid Sitobion avenae were collected from wheat and cocksfoot stands around Hampshire, UK, during March and April 1994. Eight wheat and eight cocksfoot aphid clones were chosen on the basis of readily distinguishable RAPD-PCR fingerprint profiles. The performances (weight, fecundity and survival) of successive generations of each of these clones were then carefully monitored in the laboratory as new generations of aphids were transferred either to winter wheat or to cocksfoot in planned sequences. Even those clones that were originally caught on the same host showed significant variability in performance. Clones generally performed better on their host of origin than they did on the alternate host, and they performed less well on the alternate host compared to the clones that had originated there. A comparison of the performance of third generation aphids with first generation aphids showed that the experience of the mother in the second generation often influences the subsequent performance of their offspring. As the sequence of host transfers had more effect on the performance of wheat clones than cocksfoot clones, it is likely that wheat clones are more specialised, such that wheat is a satisfactory host for cocksfoot clones but not vice versa. The study provides evidence of genetic variation in performance on host and evidence for clonal adaptation to particular host species. This adaptation may well be a major cause of the observed consistent genetic differentiability of populations of S. avenae found on wheat and roadside grasses in early spring in southern England.