Collapse temperature of solutions important for lyopreservation of living cells at ambient temperature

In this study, the collapse temperature was determined using the freeze‐drying microscopy (FDM) method for a variety of cell culture medium‐based solutions (with 0.05–0.8 M trehalose) that are important for long‐term stabilization of living cells in the dry state at ambient temperature (lyopreservation) by freeze‐drying. Being consistent with what has been reported in the literature, the collapse temperature of binary water‐trehalose solutions was found to be similar to the glass transition temperature (T′_g ～ ?30°C) of the maximally freeze‐concentrated trehalose solution (～80 wt% trehalose) during the freezing step of freeze‐drying, regardless of the initial concentration of trehalose. However, the effect of the initial trehalose concentration on the collapse temperature of the cell culture medium‐based trehalose solutions was identified to be much more significant, particularly when the trehalose concentration is less than 0.2 M (the collapse temperature can be as low as ?65°C). We also determined that cell density from 1 to 10 million cells/mL and ice seeding at high subzero temperatures (?4 and ?7°C) have negligible impact on the solution collapse temperature. However, ice seeding does significantly affect the ice crystal morphology formed during the freezing step and therefore the drying rate. Finally, bulking agents (mannitol) could significantly affect the collapse temperature only when trehalose concentration is low (<0.2 M). However, improving the collapse temperature by using a high concentration of trehalose might be preferred to the addition of bulking agents in the solutions for freeze‐drying of living cells. We further confirmed the applicability of the collapse temperature measured with small‐scale (2 µL) samples using the FDM system to freeze‐drying of large‐scale (1 mL) samples using scanning electron microscopy (SEM) data. Taken together, the results reported in this study should provide useful guidance to the development of optimal freeze‐drying protocols for lyopreservation of living cells at ambient temperature for easy maintenance and convenient wide distribution to end users, which is important to the eventual success of modern cell‐based medicine. Biotechnol. Bioeng. 2010;106: 247–259. © 2010 Wiley Periodicals, Inc.     

Predation ofOligonychus pratensis [Aca.: Tetranychidae] byPhytoseiulus persimilis andAmblyseius californicus [Aca.: Phytoseiidae] under controlled laboratory conditions

The predator efficacy ofPhytoseiulus persimilis Athias-Henriot andAmblyseius californicus (McGregor) (Acarina: Phytoseiidae) when feeding on the Banks grass mite (BGM),Oligonychus pratensis (Banks) (Tetranychidae), was compared under controlled laboratory conditions. Predation byP. persimilis andA. californicus reduced the BGM densities by 60% and 28%, respectively. In general, phytoseiids preferentially fed upon the more abundant instars. Ovipositional rates forP. persimilis while feeding on BGM approximated rates when feeding onTetranychus spp. The use of a trade name is not an endorsement by Texas A&M University.     

Effect of different release rates ofPhytoseiulus persimilis [Acarina: Phytoseiidae] on the twospotted spider mite on strawberry in southern California

The effect of different release rates of a predaceous mite,Phytoseiulus persimilis Athias-Henriot, on the twospotted spider mite,Tetranychus urticae Koch, on strawberry was investigated in southern California in 1970–71. The spider mite population reached a peak of 206.3 active stages/leaflet in the low-rate (5/plant) release plot on May 4 and 169.1/leaflet in the high-rate (10/plant) release plot on May 18, compared to a peak of 433.4/leaflet in the control (no release) plot on May 11. The mean numbers of spider mites in the low-, and high-rate release plots were significantly different from the control from April 6 through June 15, but were never significantly different from each other. The predator population reached a peak of 13.1 and 23.8 active stages/leaflet in the low-, and high-rate release plots, respectively, on May 25 which was 3 weeks and 1 week after the spider mite population reached its peak in these plots. The seasonal mean numbers of spider mite active stages in the low-, and high-rate release plots were significantly different (P=0.05) from the control, but were not significantly different from each other. There were no significant differences in fruit yield (g/plant) or fruit size between the low-, and high-rate release and control plots.

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Résumé Des lachers de l'acarien prédateurPhytoseiulus persimilis Athias-Henriot ont été réalisés en 1970–71 dans deux fraiseraies de la Californie du Sud infestées parTetranychus urticae Koch. La densité des populations larvaires deT. urticae a atteint le 4 mai un maximum de 206,3 par foliole là où le lacher avait été de 5 par plante (parcelle 1) et de 169,1 pour 10 prédateurs par plant (parcelle 2) le 11 mai, alors que le témoin en contenait 433,4 par foliole. La densité des populations larvaires deT. urticae était comparable dans les parcelles 1 et 2 entre le 6 avril et le 15 juin, mais divergeait sensiblement de celle du témoin. Les populations de larves du prédateur ont atteint un maximum de 13,1 et 23,8 par foliole dans les parcelles 1 et 2 les 13 et 25 mai, respectivement, soit 3 semaines et 1 semaine après que celles deT. urticae aient atteint leur maximum. Pour la saison, la densité moyenne des populations larvaires deT. urticae enregistrée dans les parcelles 1 et 2 différaient considérablement de celle du témoin (P=0.05), mais la différence entre les terrains 1 et 2 n'était pas significative. Il n'y a pas eu de différences importantes entre la récolte de fraises (g/plante) ou la dimension des fruits récoltés dans les parcelles 1 et 2 et le témoin.

     

Constraints to the implementation of biological control in Sri Lanka

Sri Lanka is a tropical island with a rich diversity of arthropods, with many species of indigenous natural enemies of economic pests. However, no extensive island wide surveys have been carried out for natural enemies of major crop ecosystems and only a few of the indigenous natural enemies are reported useful for augmentation biological pest control by massive field releases of laboratory-produced insects. Most successful biological control programs on the island are importation (i.e. classical) biological control programs, where established exotic natural enemies control certain pest populations in valuable crops. There is growing interest in the use of native natural enemies in pest management, thus creating a need for intensive research on the ecology of indigenous natural enemies and development of infrastructure and technology to produce natural enemies for augmentation. This paper examines constraints and opportunities for implementation of biological control in Sri Lanka.     

Response of an aphid parasitoid, Aphelinus asychis to its host, plant, host-plant complex, and to malathion

Aphelinus asychis (Walker) can be valuable as a biocontrol agent of the aphid, Schizaphis graminum (Rondani), a major pest on grain crops in the United States. This study reports the wind tunnel, and olfactometric responses of this parasitoid to the host (aphid), plant (sorghum), and host-plant complex (aphid-infested sorghum). In addition, the parasitoids' response to malathion-treated plants is also presented. The goal of the present study was to test the hypothesis that volatiles associated with the host attract natural enemies, as reported in cases of many hymenopterous parasitoids, and also to learn about the effects of insecticides on these parasitoids. In wind tunnel studies, these parasitoids moved upwind in the direction of the host-infested plant. There was no direct flight observed, however, these parasitoids were observed to hop and jump, and sometimes walk to their host. In the olfactometer experiments, we found that A. asychis is attracted to host-plant complex. The parasitoids' response to malathion in the olfactometer suggested that a malathion formulation when applied to plants can lure these beneficials, thus providing new insight into the ongoing task of integrating chemical and biological control of insect pests.     

Braconid parasitoids ofTephritidae [Diptera] infesting coffee and other fruits in West-Central Africa

Exploration for parasitoids ofCeratitis capitata (Wiedemann) [Diptera: Tephritidae] was conducted in Cameroon and Togo and parasitoids collected were released in Costa Rica. Collections yielded large numbers ofOpius perproximus Silvestri,Biosteres caudatus Szepligeti, andB. caudatus auct. [Opiinae: Braconidae]. Other Opiines collected wereB. fullawayi (Silvestri),B. desideratus (Bridwell), and an undescribed species ofOpius. Parasitization of tephritids in coffee ranged from 10–56%, with an average 35% parasitism in a research plantation and 17% in a commercial plantation. Parasitism in fruits other than coffee was less than 5%. The most commonly reared tephritids from coffee wereTrirhithrum coffeae Bezzi andC. (Pterandrus) anonae Graham.C. capitata occurred in low frequency in coffee; and adults were rarely observed. Other rearings includedT. albonigrum (Enderlein),T. sp. nr.validum Bezzi,T. nigerrimum auct. nec. Bezzi,C. (Pterandrus) colae Silvestri,C. (Pterandrus) flexuosa (Walker),C. (Ceratalaspis) spp.cosyra (Walker) group,C. (Pardalaspis) sp.punctata (Wiedemann) group,C. (Ceratalaspis) giffardi Bezzi,Dacus (Dacus) bivittatus (Bigot),D. (Didacus) ciliatus Loew, andGymnodacus sp.

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Résumé Une récolte des parasites deCeratitis capitata (Wiedemann) [Diptères: Téphritidés] a été conduite au Cameroun et au Togo, et les specimens, recueillis ont été libérés au Costa Rica. Cette étude a mis, en évidence la présence d'un grand nombre d'Opius perproximus Silvestri, deBiosteres caudatus Szépligeti, et deB. caudatus auct. (Braconidés: Opiinés). Les autres opiinés collectés comprenaientB. fullawayi (Silvestri),B. desideratus (Bridwell), et une espèce non décrite du genreOpius. Le taux de parasitisme des téphritidés collectées sur le café variait de 10 à 56%, avec une moyenne de 35% dans une plantation expérimentale et 17% dans une plantation commerciale. Sur les autres fruits, le taux de parasitisme était inférieur à 5%. Les espèces de téphritidés élevées le plus fréquemment des grains de café étaientTrirhithrum coffeae Bezzi etC. (Pterandrus) anonae Graham.C. capitata n'était présente qu'en faible abondance dans le café et des adultes n'ont que rarement été observés. Les autres espèces rencontrées comprenaientT. albonigrum (Enderlein),T. sp. nr.validum Bezzi,T. nigerrimum auct. nec. Bezzi,C. (Pterandrus) colae Silvestri,C. (Pterandrus) flexuosa (Walker),C. (Ceratalaspis) spp. groupecosyra (Walker),C. (Pardalaspis) sp. groupepunctata (Wiedemann),C. (Ceratalaspis) giffardi Bezzi,Dacus (Dacus) bivittatus (Bigot),D. (Didacus) ciliatus Loew, andGymnodacus sp.

     

Local and global methods of assessing thermal nociception in Drosophila larvae

In this article, we demonstrate assays to study thermal nociception in Drosophila larvae. One assay involves spatially-restricted (local) stimulation of thermal nociceptors while the second involves a wholesale (global) activation of most or all such neurons. Together, these techniques allow visualization and quantification of the behavioral functions of Drosophila nociceptive sensory neurons. The Drosophila larva is an established model system to study thermal nociception, a sensory response to potentially harmful temperatures that is evolutionarily conserved across species. The advantages of Drosophila for such studies are the relative simplicity of its nervous system and the sophistication of the genetic techniques that can be used to dissect the molecular basis of the underlying biology In Drosophila, as in all metazoans, the response to noxious thermal stimuli generally involves a "nocifensive" aversive withdrawal to the presented stimulus. Such stimuli are detected through free nerve endings or nociceptors and the amplitude of the organismal response depends on the number of nociceptors receiving the noxious stimulus. In Drosophila, it is the class IV dendritic arborization sensory neurons that detect noxious thermal and mechanical stimuli in addition to their recently discovered role as photoreceptors. These neurons, which have been very well studied at the developmental level, arborize over the barrier epidermal sheet and make contacts with nearly all epidermal cells. The single axon of each class IV neuron projects into the ventral nerve cord of the central nervous system where they may connect to second-order neurons that project to the brain. Under baseline conditions, nociceptive sensory neurons will not fire until a relatively high threshold is reached. The assays described here allow the investigator to quantify baseline behavioral responses or, presumably, the sensitization that ensues following tissue damage. Each assay provokes distinct but related locomotory behavioral responses to noxious thermal stimuli and permits the researcher to visualize and quantify various aspects of thermal nociception in Drosophila larvae. The assays can be applied to larvae of desired genotypes or to larvae raised under different environmental conditions that might impact nociception. Since thermal nociception is conserved across species, the findings gleaned from genetic dissection in Drosophila will likely inform our understanding of thermal nociception in other species, including vertebrates.     

Hymenopterous EGG-pupal and larval-pupal parasitoids ofCeratitis capitata andAnastrepha spp. [Dip.: Tephritidae] in Costa Rica

Egg-pupal and larval-pupal parasitoids were recovered from less than 10% of the 16,000 tephritid puparia collected in Costa Rica from August, 1979, through April, 1980.Ceratitis capitata (Wiedemann) was attacked by 2 introduced opiineBraconidae and 2 indigenous eucoilineCynipidae Anastrepha spp. were attacked by each parasitoid species attackingC. capitata and also by 5 indigenous opiineBraconidae and 1 exoticEulophidae. Toxotrypana curvicauda Gerstaecker was attacked only by an indigenous opiineBraconidae which did not attack other tephritid species collected.     

Biology and life tables for the predaceous earwig,Doru taeniatum [Derm.: Forficulidae]

The biology of the earwig,Doru taeniatum (Dohrn), a reported predator ofSpodoptera frugiperda (J. E. Smith), was studied in the laboratory.D. taeniatum under laboratory conditions was easily reared, mated readily and females tended eggs and first instar nymphs.D. taeniatum developed through 4 nymphal instars and reached imago in approximately 30 days. It is an omnivore in the laboratory and capable of developing and reproduction on plant pollen alone, and on Lepidoptera eggs (Sitotroga carealella) (Olivier) alone. Immatures developed most rapidly and females were most fecund when fed either a combination ofS. cereallea eggs and pollen, orS. cerealella eggs alone. Approved as TA 20744 by Director, Texas Agricultural Experiment Station.