Patterns of primary spore discharge of Entomophthora spp from the blue green aphid,Acyrthosiphon kondoi

Experiments were conducted to study the effects of time, temperature, and light regime on primary spore formation at 100% RH for the three major pathogens of Acyrothosiphon kondoi. Only small differences were detected between the continuous light and continuous dark regimes. Entomophthora obscura produced between 6 and 10 × 10³ primary spores mostly during the first 48 hr. Total primary spore production was similar at the five temperatures tested from 5° to 25°C. Entomophthora planchoniana produced large numbers of primary spores (about 5 × 10⁴ per aphid) only at temperatures between 10° and 20°C. The majority of primary spores were formed during the first 24 hr. Primary spore production with Entomophthora nr. exitialis ranged from about 10⁵ per aphid at 5° and 10°C to 3 or 4 × 10⁵ at 15° to 25°C, with most spores being formed during the first 48 hr. It is suggested that rainfall is more likely to be important for transmission of E. obscura and E. nr. exitialis than for transmission of E. planchoniana, and that E. obscura is likely to be the most important pathogen during cool or cold weather.     

The survival of Entomophthora spp. in mummified aphids at different temperatures and humidities

Entomophthora aphidis survived for at least 32 weeks at 0°C and 20 or 50% RH and E. thaxteriana for at least 16 weeks at 10°C and 20 or 50% RH in mummified infected pea aphids, Acyrthosiphon pisum. The fungi produced infective conidia when the aphids were moistened. This probably explains the survival of Entomophthora species infecting aphids during short periods when the weather is unsuitable for conidial discharge and host infection.     

Effect of temperature on the life history of <Emphasis Type="Italic">Eretmocerus</Emphasis> sp. nr. <Emphasis Type="Italic">furuhashii</Emphasis>, a parasitoid of <Emphasis Type="Italic">Bemisia tabaci</Emphasis>

Eretmocerus sp. nr. furuhashii (Hymenoptera: Aphelinidae) is an indigenous parasitoid of Bemisia tabaci (Gennadius)(Hemiptera: Aleyrodidae) from southern China; the effects of constant temperatures on the life history of E. sp. nr. furuhashii were examined in the laboratory. The developmental period ranged from 39.2 days at 20°C to 12.40 days at 32°C. A total of 263.4 degree-days were required to complete development with a lower developmental threshold temperature of 11.1°C. Of the eggs produced, 59.3% completed development at 20°C with completion increasing to 71.5% at 26°C. Adult female longevity was 10.8 days at 20°C and 5.2 days at 32°C while the mean daily offspring reproduced per female was highest at 29°C with 5.9 offspring. Adult oviposition peaked three days after emergence at 26, 29 and 32°C, and four days post-emergence at 20°C and 23°C. The total numbers of offspring produced per female ranged from 25.7 individuals at 32°C to 41.1 individuals at 20°C. The sex ratio had a female bias and ranged from 0.72 at 17°C to 0.51 at 35°C. The intrinsic rate of increase was 0.1727 at 29°C followed with 0.1606 at 32°C. Results indicated that E. sp. nr. furuhashii reaches its maximum biological potential at temperatures ranging from 26°C to 32°C.     

Mortality of bifidobacteria in boiled yogurt

Bifidobacterial strains showed various mortalities during storage at 30°C in boiled yogurt prepared with Streptococcus sp. and Lactobacillus sp. Bifidobacterium breve 203 was most stable, maintaining its initial cell number for more than 5 days. It was also stable at 4 or 10°C in fresh yogurt. B. longum 401 rapidly lost viability in the boiled yogurt and was unstable in the fresh yogurt at any temperature. Lactobacillus sp. remained fully viable for one week or more at 4–30°C while Streptococcus sp. lost viability at 20°C or above. The difference in mortality between B. breve 203 and B. longum 401 was mainly due to their sensitivity to the acidic environment, with temperature during storage having a secondary effect. Effects of lysozyme, pepsin and bile acids on the two strains were also investigated.     

Efficacy of the Ventral Abdominal Secretion of the Cockroach Eurycotis floridana (Blattaria: Blattidae) as a Defense Allomone

In assays investigating the abdominal sternal secretion of the cockroach Eurycotis floridana, it was determined that the secretion provides an effective deterrent against potential predators. Analysis of grooming patterns of Peromyscus leucopus mice revealed significant increases in mouth grooming and loss of coordination following exposure to E. floridana secretion. Mouse–cockroach interactions also changed following exposure of the mouse to the spray, as the mouse incited fewer interactions and more frequently struck with its forepaws than with its mouth. Monomorium sp. and Camponotus sp. ants also were repelled by the secretion, and Periplaneta americana were significantly irritated by the application of E. floridana secretion. The secretion is able to deter without physical contact, presumably via noxious volatiles. Eurycotis floridana is capable of accurately aiming the discharge as well as ejecting it several times the length of its body. It also was found that E. floridana exhibits autotoxic responses following contact with the secretion of conspecifics.     

Survival of anaerobic fungus Caecomyces sp. in various preservation methods: a comparative study

The present investigation was designed to observe the survival of the anaerobic fungus Caecomyces sp. in various routine preservation methods. Among all the treatments, cryopreservation of fungi at ?70°C with glycerol was found to be most effective for long-term maintenance (more than 90 days) of rumen fungi, followed by dimethyl sulfoxide (DMSO) and ethylene glycol (up to 60 days). In contrast, at ?196°C, DMSO showed maximum survival (more than 90 days), followed by glycerol (up to 90 days) and ethylene glycol (up to 30 days). At 39°C, maximum survival (up to 30 days) was observed with soft agar and wheat straw; at refrigeration temperature, preservation with Orpin's media containing straw showed maximum survival (up to 30 days).     

Continuous production of l-phenylalanine from acetamidocinnamic acid using co-immobilized cells of Corynebacterium sp. and Paracoccus denitrificans

In order to produce l-phenylalanine efficiently from acetamidocinnamic acid with immobilized microbial cells, a two-step enzyme reaction using the acetamidocinnamate amidohydrolase activity of Corynebacterium sp. C-23 cells and the aminotransferase activity of Paracoccus denitrificans pFPr-1 cells was investigated. It was found that the useage of co-immobilized Corynebacterium sp. and P. denitrificans cells with κ-carrageenan was superior to that of the mixture of immobilized Corynebacterium sp. cells and immobilized P. denitrificans cells. When the space velocity was 0.06 h⁻¹ at 30°C, 147 mml-phenylalanine were produced with a 98% conversion ratio from acetamidocinnamic acid. The half-life of the l-phenylalanine-forming activity of the column was calculated to be ≈ 14 days at 30°C.     

Characterization of a family 3 polysaccharide lyase with broad temperature adaptability, thermo-alkali stability, and ethanol tolerance

The 774-bp pectate lyase gene plyAI4 from Bacillus sp. I4 was cloned and expressed in E. coli. The gene encodes a 257-residue polypeptide (PlyAI4, 28.3 kDa) with the highest identities of 97.3% with a putative pectate lyase from Bacillus subtilis BSn5 (ADV94306) and 60.3% with an identified pectate lyase of the polysaccharide lyase family (PL) 3 from Paenibacillus amylolyticus 27C64 (ADB78774). The purified recombinant PlyAI4 (rPlyAI4) exhibited apparently optimal activity at pH 10.5 ?? 11.0 and 50°C. Compared with the majority of reported alkaline pectate lyases, rPlyAI4 exhibited more residual enzyme activity at 20°C (??45%) or at 70°C (??50%) and better thermostability at 70°C (??60 min half-life at 70°C). In the presence of 20% (v/v) ethanol, pectate lyase activity was enhanced by 0.2 fold. After incubation in 40% (v/v) ethanol at 37°C and pH 8.5 for 1 h, the purified rPelAI4 retained more than 75% of the initial activity. Sequence analysis proposed a new signature block, A-D-G-[V/I]-H, for PL 3 pectate lyases. These properties may prove to be important with regards to PlyAI4 for basic research and industrial application.     

Thermotolerant oil-degrading bacteria isolated from soil and water of geographically distant regions

Oil-degrading bacteria were isolated from soil and water samples taken in Russia, Kazakhstan, and the Antarctic; 13 of 86 strains proved to be thermotolerant. These bacteria utilized crude oil at 45–50°C; their growth optimum (35–37°C) and range (20–53°C) differ from those of mesophilic bacteria. Thermotolerant strains were identified as representatives of the genera Rhodococcus and Gordonia. It was shown that their ability to degrade petroleum products does not differ at 24 and 45°C. The strains Rhodococcus sp. Par7 and Gordonia sp. 1D utilized 14 and 20% of the oil, respectively, in 14 days at 45°C. All of the isolated thermotolerant bacteria grew in a medium containing 3% NaCl; the medium for the strains Gordonia amicalis 1B and Gordonia sp. 1D contained up to 10% NaCl. The bacteria G. amicalis and Rhodococcus erythropolis were able to utilize crude oil and individual hydrocarbons at higher (up to 50°C) temperatures.     

Observations on the morphology and infectivity for cattle of Babesia bovis parasites in unfed Boophilus microplus larvae after incubation at various temperatures

Dalgliesh R. J. and Stewart N. P. 1979. Observations on the morphology and infectivity for cattle of Babesia bovis parasites in unfed Boophilus microplus larvae after incubation at various temperatures. International Journal for Parasitology9: 115–120. The temperature of incubation of unfed Boophilus microplus larvae infected with Babesia bovis influenced the morphology and infectivity of the Babesia within the tick. Incubation at 37°C for 1–3 days stimulated the development of parasites morphologically similar to those usually observed in fed larvae harvested from cattle; similar forms appeared more slowly in larvae incubated at 31°C or 25°C. Extracts prepared from larvae after incubation at 37°C for 3–5 days or 30°C for 8 days were consistently infective for cattle. Prior storage of larvae at 14°C for up to 28 days enhanced the development of infectivity at 37°C; infectivity could still be produced after 65 days storage at 14°C but not after 76 days. Larvae released on a host transmitted B. bovis sooner if they had been incubated at 37°C for 4 days. It was concluded that the development of B. bovis to an infective stage in B. microplus is temperature dependent and does not require the stimulus of feeding by the host.     

Effect of temperature on germination,growth, and infectivity of the mosquito pathogen Tolypocladium cylindrosporum (deuteromycotina; hyphomycetes)

The mosquito pathogen Tolypocladium cylindrosporum was examined with regard to its response to temperature. Similar temperature ranges were found for growth, germination, and infectivity of blastospores and conidia. Germination occurred at 8° and 33°C but not at 6° and 35°C. Optimal germination and growth was noted between 24° and 27°C for both spore types. Infectivity of blastospores and conidia at different temperatures was examined by exposing L₂Aedes sierrensis larvae to concentrations of 5 × 10⁵ blastospores/ml or 5 × 10⁶ conidia/ml. Larvae were incubated at 12°, 15°, 25°, and 30°C. Infection occurred at all temperatures tested with LT₅₀ values ranging from 22.7 days (12°C) to 5.6 (25°C) days for conidia and 4.7 days (12°C) to 0.6 day (25°C) for blastospores. These results confirmed earlier findings that blastospores infected and killed host larvae more rapidly than conidia and suggested that this difference is largely due to the more rapid germination rate of blastospores. These experiments demonstrated that T. cylindrosporum can be active against mosquito larvae over a broad range of temperatures encompassing both the cold-water habitat of certain temperate mosquito species as well as the habitat of tropical vector species.     

Differential Responses of the Whitefly Bemisia tabaci Symbionts to Unfavorable Low and High Temperatures

The whitefly Bemisia tabaci complex contains many cryptic species, of which the Middle East-Asia Minor 1 (MEAM1) and Mediterranean (MED) are notorious invasive pests. In our field-collected whitefly samples, MEAM1 harbors an obligate primary symbiont “Candidatus Portiera aleyrodidarum” and two secondary symbionts, “Candidatus Hamiltonella defensa” and Rickettsia sp., whereas MED has only “Ca. Portiera aleyrodidarum” and “Ca. Hamiltonella defensa.” Both “Ca. Portiera aleyrodidarum” and “Ca. Hamiltonella defensa” are intracellular endosymbionts residing in the bacteriomes, whereas Rickettsia sp. has a scattered distribution throughout the host body cavity. We examined responses of these symbionts to adverse temperatures as well as survival of the host insects. After cold treatment at 5 or 10 °C or heat treatment at 35 or 40 °C for 24 h, respectively, the infection rates of all symbionts were not significantly decreased based on diagnosis PCR. However, quantitative PCR assays indicated significant reduction of “Ca. Hamiltonella defensa” at 40 °C, and the reduction became greater as the duration increased. Compared with “Ca. Hamiltonella defensa,” “Ca. Portiera aleyrodidarum” was initially less affected in the first day but then showed more rapid reduction at days 3–5. The density of Rickettsia sp. fluctuated but was not reduced significantly at 40 °C. Meanwhile, the mortality rates of the host whiteflies elevated rapidly as the duration of exposure to heat treatment increased. The differential responses of various symbionts to adverse temperatures imply complex interactions among the symbionts inside the same host insect and highlight the importance of taking the whole bacterial community into account in studies of symbioses.     

Studies on Entomophthora in populations of Aphis fabae on field beans

The population of Aphis fabae on field beans at a site in Highfield, Rothamsted in 1973 reached its peak 1 wk earlier than that at an equivalent site in Mill Dam Close, Woburn, 29 km NW of Rothamsted. Epizootics of Entomophthora caused weekly maximum mortalities of adult apterae of 71% at Highfield and 67% at Mill Dam Close. These epizootics and the periodicity of Entomophthora conidia in the air closely paralleled the development of the aphid population. There was little evidence of a consistent relationship between Entomophthora infection and any of the weather factors considered. At both sites most mortality was caused by E. planchoniana though many aphids were killed by E. aphidis and E. obscura. E. fresenii and E. virulenta killed very few aphids. Most conidia in the air were of the E. aphidis-type. Up to 44% of alatae emigrating from bean crops were infected with Entomophthora, confirming that aphid migration is an important means of distributing the fungi. Aphid numbers rose to more than 1600/plant at both sites, in spite of the action of Entomophthora, and would probably have been less had the fungi been more abundant earlier in the season.     

Gene cloning,purification, and characterization of the highly thermostable leucine dehydrogenase of Bacillus sp.

The leucine dehydrogenase (l-leucine: NAD⁻ oxidoreductase, deaminating, EC 1.4.1.9) gene from Bacillus sp. DSM730 was cloned into Escherichia coli C600 with a vector plasmid, pBR322. The E. coli cells carrying a recombinant plasmid, pKULD1 (9.5 kb), produced a highly thermostable leucine dehydrogenase. The enzyme from E. coli cells carrying pKULD103, a deletion plasmid (6.5 kb) of pKULD1, was purified to homogeneity from the crude extract of clone cells by only one ion-change chromatography application with a yield of 73%. The leucine dehydrogenase of Bacillus sp. DSM730 is very similar in enzymological properties to those of other bacteria, except for molecular weight and stability. It has a molecular weight of about 280,000 and consists of six subunits identical in molecular weight (47,000). The enzyme is not inactivated by heat treatment at 80°C for 10 min, and incubation in the pII range of 5.4 to 10.3 at 55°C for 10 min. The Bacillus sp. DSM730 leucine dehydrogenase is the most thermostable of the leucine dehydrogenases so far purified, and is very useful for structure and stability studies, as well as being applicable to l-leucine production.     

A novel phospholipase D constitutively secreted by Ochrobactrum sp. ASAG-PL1 capable of enzymatic synthesis of phosphatidylserine

Phospholipase D (PLD) plays a crucial role in the enzyme-mediated preparation of phosphatidylserine (PS). A new PLD constitutively secreted by Ochrobactrum sp. ASAG-PL1 is now reported for the first time. After purification, the MW of the enzyme was ~37 kDa by SDS-PAGE. Its activity was highest at 40 °C and pH 7.0 and more than 90 % of the initial activity was maintained after 30 days at 4 °C and pH 7.0. This enzyme may then be useful as a potential biocatalyst for PS production.     

Effects of temperature and dosage on Vairimorpha sp. 696 spore morphometrics,spore yield,and tissue specificity in Heliothis virescens

The effects of temperature and dosage on a new microsporidian species, Vairimorpha sp. 696, were examined in H. virescens. The pathogen was evaluated for tissue specificity, spore size, cumulative percentage mortality, and spore production. All tissues examined bore infection at 32°C. Spore length was significantly longer at 19°C (5.9 μm) than at 32°C (4.7 μm). Spore widths at these two temperatures did not differ significantly. Octospores were not found at either temperature at 8 or 12 days postinoculation. One hundred percent mortality was attained in all dosages administered, but the initial rate of mortaily was more rapid in the higher dosages. Finally, spore yield was greater in larvae administered lower dosages. Maximum spore yield at 27°C was 4.87 × 10⁹ spores/larva.     

Effect of different cold storage periods on parasitization performance of Trichogramma cacoeciae (Hymenoptera,Trichogrammatidae) on eggs of Ephestia kuehniella (Lepidoptera,Pyralidae)

The parasitism rates by Trichogramma cacoeciae Marchal (Hymenoptera, Trichogrammatidae) using Ephestia kuehniella Zell. (Lepidoptera, Pyralidae) eggs held at 0, 4 and 8°C and for up to 31 days was measured. Parasitism was lowest on eggs held at 8°C and highest on eggs held at 0°C. The highest parasitism, 97.8%, was measured for parasitoids attacking eggs held for 3 days and stored at 0°C. Parasitism of eggs stored at all three temperatures decreased with increasing duration of storage. The number of T. cacoeciae successfully developing and emerging as adults after storage in E. kuehniella eggs held at 0, 4 and 8°C was measured. Parasitoid emergence was >83% from E. kuehniella eggs stored at 8°C for 3 weeks. Storage at 0°C caused a significant decline in parasitoid emergence after 2 weeks (P<0.05). Storage at 0°C for more than 4 weeks reduced fecundity by 50%. T. cacoeciae parasitized the highest number of E. kuehniella eggs 1 day after adult emergence. The oviposition period lasted 6–7 days, although the parasitoids lived up to 13–14 days. Impact of storage time and temperature on parasitism rates by T. cacoeciae stored while in E. kuehniella eggs was measured. As storage time and temperature increased, subsequent parasitism rates of resulting adult T. cacoeciae decreased. Eggs of E. kuehniella can be stored at 0°C for up to 31 days. Trichogramma cacoeciae developing in eggs of E. kuehniella can be stored at 4°C for up to 5 weeks prior to release.     

Quantitative studies on the uptake and retention of potato leafroll virus by aphids in laboratory and field conditions

Enzyme-linked immunosorbent assay (ELISA) was adapted for the efficient detection and assay of potato leafroll virus (PLRV) in aphids. Best results were obtained when aphids were extracted in 0.05 M phosphate buffer, pH 7.0, and the extracts incubated at 37 °C for 1 h before starting the assay. Using batches of 20 green peach aphids (Myzus persicae), about 0.01 ng PLRV/aphid could be detected. The virus could also be detected in single aphids allowed a 1-day acquisition access period on infected potato leaves. The PLRV content of aphids depended on the age of potato source-plants and the position of source leaves on them. It increased with increase in acquisition access period up to 7 days but differed considerably between individual aphids. A maximum of 7 ng PLRV/aphid was recorded but aphids more usually accumulated about 0.2 ng PLRV per day. When aphids were allowed acquisition access periods of 1–3 days, and then caged singly on Physalis floridana seedlings for 3 days, the PLRV content of each aphid, measured subsequently, was not strongly correlated with the infection of P. floridana. The concentration of PLRV in leaf extracts differed only slightly when potato plants were kept at 15, 20, 25 or 30 °C for 1 or 2 wk, but the virus content of aphids kept on leaves at the different temperatures decreased with increase of temperature. PLRV was transmitted readily to P. floridana at all temperatures, but by a slightly smaller proportion of aphids, and after a longer latent period, at 15 °C than at 30 °C. The PLRV content of M. persicae fed on infected potato leaves decreased with increasing time after transfer to turnip (immune to PLRV). The decrease occurred in two phases, the first rapid and the second very slow. In the first phase the decrease was faster, briefer and greater at 25 and 30 °C than at 15 and 20 °C. No evidence was obtained that PLRV multiplies in M. persicae. These results are compatible with a model in which much of the PLRV in aphids during the second phase is in the haemocoele, and transmission is mainly limited by the rate of passage of virus particles from haemolymph to saliva. The potato aphid, Macrosiphum euphorbiae, transmitted PLRV much less efficiently than M. persicae. Its inefficiency as a vector could not be ascribed to failure to acquire or retain PLRV, or to the degradation of virus particles in the aphid. Probably only few PLRV particles pass from the haemolymph to saliva in this species. The virus content of M. euphorbiae collected from PLRV-infected potato plants in the field increased from early June to early July, and then decreased. PLRV was detected both in spring migrants collected from the plants and in summer migrants caught in yellow water-traps. PLRV was also detected in M. persicae collected from infected plants in July and August, and in trapped summer migrants, but their PLRV content was less than that of M. euphorbiae, and in some instances was too small for unequivocal detection.     

Laboratory studies on the life-histories of four enchytraeid worms (Oligochaeta) which inhabit sewage percolating filters

The life-histories of four enchytraeid worms, Lumbricillus rivalis, Enchy-traeus coronatus, E. buchholzi, and E. albidus which occur in sewage percolating filters, were studied under laboratory conditions at 8 , 15 and 20°C. The number of ova per cocoon varied from 0 to 50 (L. rivalis), 0 to 33 (E. coronatus), 1 to 9 (E. buchholzi) and 0 to 22 (E. albidus). The mean number of ova per cocoon was highest at 15°C for all species except E. coronatus which had a highest mean value at 8°C. The number of ova in cocoons was correlated with cocoon length (P < 0.001) for all species. Cocoon production usually increased with temperature ranging from 0.8 cocoons per adult per week at 8°C to 2.0 at 20°C for L. rivalis, and from 1–4 to about 2.6 for E. coronatus and E. buchholzi. The total number of ova produced by each E. coronatus (350 at 8°C to 550 at 20°C) was similar to that produced by each L. rivalis (600 at 8°C to 350 at 20°C) and was about five times greater than the total numbers produced by the other two species. Cocoon and ova production and the number of ova per cocoon varied with the age of the adult, usually reaching a peak soon after maturity. Hatching success was low and generally 40–50 % of ova failed to develop; subsequent mortality among immature worms was about 10–20%. Growth was more rapid at the higher temperatures; L. rivalis matured in about 26 days at 20°C, the clitellum forming when the worm was 13–14 mm long; data for the other species are 13 days and 5–6 mm (E. coronatus); 16 days and 3–4 mm (E. buchholzi); 28 days and 13–14 mm (E. albidus). The maturation period at 8°C was at least twice that at 20°C. The generation period (cocoon to cocoon) was about a month at 20°C for all species except E. albidus (2 months), but as some species had longer reproductive periods than others the actual number of generations per year was highest in E. buchholzi, 7.0 per year, and lowest in E. albidus, about 3.3 per year, At 8°C all four species had between 1.4 and 2.8 generations a year. A comparison of expected and observed population densities of L. rivalis and E. coronatus in a sewage percolating filter showed that neither achieved values approaching their potential summer densities although ample food was apparently available. Of the four species studied only E. buchholzi produced viable ova without pairing.     

Development rate,number of mature oocytes at emergence and adult size ofEncarsia tricolor at constant and variable temperatures

The duration of the development of the aphelinidEncarsia tricolor Föerster (a parasitoid of the aleyrodidTrialeurodes vaporariorum (Westwood), adult size and number of mature oocytes at emergence were determined under constant and variable temperature regimes. Females developed successfully from 14 to 32°C, but a 100% of pupal mortality was observed at 34°C. Males developed successfully from 16 to 28°C and they developed faster than females. Female and male, egg to adult development at constant temperatures ranged from 51.1 (14°C) to 14.3 (28°C) days and from 32.6 (16°C) to 11.8 (28°C) days, respectively. Predictions of the rate of development at variable temperatures were more accurate when made from 2nd and 3rd degree polynomials than from linear regressions. The number of mature oocytes at emergence ranged from 0.1 (30°C) to 2.2 (20°C). FemaleE. tricolor attained the greatest size at 20–24°C. The comparison with literature data shows thatE. tricolor develops faster thanT. vaporariorum at temperatures above 15°C.