The influence of previous host age on current host acceptance in Trichogramma

Trichogramma principium Sug. & Sor. females were sequentially offered two portions of the grain moth (Sitotroga cerealella Oliv.) eggs, either young (1-day old) or old (eggs that had developed 6 days at a temperature of 20 °C). The probability of host acceptance depended not only on current host age, but also on the age of the previously offered host. Particularly, Trichogramma females more often oviposited in old host eggs when previously offered young eggs (35–45% of Trichogramma females laid eggs) compared to females which were sequentially offered two portions of old eggs (15–20% of Trichogramma females laid eggs). In other words, parasitization by Trichogramma was stable even when transferred from young (preferred) to old (usually rejected) eggs. Dissections showed that refusing females had significantly more mature eggs than ovipositing females, independent of host age. Among ovipositing females, wasps provided with young hosts had fewer mature ovarial eggs than wasps provided with old hosts. Supposedly, Trichogramma females offered old hosts require a higher motivation to oviposit and have a correspondingly higher egg load than females offered young (preferred) hosts.     

Enhanced (Na+K)-ATPase Activity and Expression in Mouse Brain after Chronic Ethanol Administration

It is the general hypothesis that the primary mode of action of ethanol is the alteration of membrane structure and function including the conformation of receptors and ion channels essential for neurotransmission and signal transduction. However, the issue of whether ethanol affects (Na+K)-ATPase under physiological conditions remains unsettled. In this study, adult mice were treated with a daily dose of 5 g/kg of ethanol for 28 days. The RNA was isolated from brain and the (Na+K)-ATPase mRNA level was determined using Northern blot analysis. We have found an increased expression of (Na+K)-ATPase -subunit in the chronically treated alcohol group as compared with that of controls. This result was further substantiated by increased protein phosphorylation as well as increased specific activity of this enzyme in the synaptosomal plasma membrane after chronic ethanol administration. Thus we have demonstrated that ethanol may directly affect (Na+K)-ATPase in vivo, leading to the increased synthesis of this enzyme through adaptive mechanisms.     

Chemical signals mediating interactions betweenGaleruca tanaceti L. (Coleoptera, Chrysomelidae) and its egg parasitoidOomyzus galerucivorus (Hedqvits) (Hymenoptera, Eulophidae)

Chemical signals mediating interactions betweenGaleruca tanaceti and its egg parasitoidOomyzus galerucivorus (Hymenoptera, Eulophidae) were studied. Neither odor of gravid females ofG. tanaceti nor volatiles of their feces were attractive to the parasitoid. However, the presence of the beetles’ feces on a substrate arrested the parasitoid and elicited frequent antennal drumming. Thus, feces may contain a kairomone important for host finding. Odors of damaged and undamaged host plants had no effect on the parasitoids.O. galerucivorus did not detect its host eggs at close range but encountered them by chance. Neither the structure nor the dark color of the egg surface play a key role in host recognition, but chemicals of the extrachorion which could be isolated by dichloromethane. Fractionation of the dichloromethane extract by TLC revealed a single active fraction which induced host recognition behavior. Since the eggs ofG. tanaceti contain anthraquinones and anthrones which are active as feeding deterrents against predators, we hypothesized that reproductive success ofO. galerucivorus is due to sequestration of these protective compounds. However, GC-MS analyses revealed that there was no transfer of them from the host egg into the adult parasitoid.     

Phenotypic characteristics and composition of rhizobia associated with woody legumes growing in diverse Kenyan conditions

Over 480 rhizobia were isolated from root nodules of woody legume and herbaceous trap host species grown in soils collected from 12 different Kenyan sites. The isolates were differentiated by growth and morphological characteristics, intrinsic antibiotic resistance (IAR) and salt (NaCl) tolerance levels (STL) when grown on yeast mannitol mineral salts agar and broth media.The bulk of the isolates (91%) were watery, milky-translucent and curdled milk types with moderate to copious extracellular polysaccharide (EPS). The rest were creamy or white opaque with little to moderate EPS production. Overall, they showed a wide range of growth rates: very fast-growing (mean generation time 1.6–2.5 h), fast-growing (2.8–4.8 h), intermediate between fast- and slow-growing (5.6–5.7 h) and slow- and very slow-growing (6.4–8.8 h). The isolates were tentatively grouped into Rhizobium spp., to include very fast, fast and intermediate (acid-producing) types; and Bradyrhizobium spp., to include very slow, slow and intermediate (alkali-producing) types.Bradyrhizobium spp. were more sensitive to antibiotics (40 g mL^-1) than Rhizobium spp., contrary to the general opinion which indicates that they are normally resistant. Cluster analysis based on sensitivity responses of IAR and STL could not distinguish Rhizobium spp. from Bradyrhizobium spp., neither was there any association by site nor host of isolation except for those isolates trapped with Phaseolus vulgaris at Kibwezi.Our data demonstrated a high diversity of tropical rhizobia associated with trees.     

Effect ofEscherichia coliIHF Mutations on Plasmid p15A Copy Number

Four isogenic strains (himAhimDdouble mutant,himAandhimDsingle mutants, and their wild type counterpart) harboringorip15A plasmid (pACYC184 or pACYC184Amp or pACYC177) show different copy numbers of that plasmid in the early stationary phase of cultivation. The copy number oforip15A plasmid increases about four times in thehimAhimDdouble (65–70 copies per cell) andhimDsingle mutant cells (50–56 copies per cell) and was almost the same inhimAmutant (17–18 copies per cell) and wild type cells (14–16 copies per cell). The results suggest that HimD can form homodimers, which are functionally competent for the regulation oforip15A plasmid copy number. Complementation experiments ofhimAhimDdouble mutant cells using plasmid carryinghimAandhimDgenes (pP_LhiphimA-5) confirm the effect of integration host factor (IHF) absence on increasing the copy number oforip15A plasmid (plasmid producing IHF complemented the defect of IHF mutant). The absence of IHF (usinghimAhimDdouble mutant as host) had no effect on the copy number of the pBR322 (oripMB1) plasmid.     

Differences in ecological and genetic traits between Panonychus citri (Acari: Tetranychidae) populations infesting citrus and Osmant hus

Comparative studies of the ecological and genetic traits of Panonychus citri (McGregor) populations on citrus and other host plants were conducted. Panonychus citri on Osmanthus trees has a unique esterase allele and host range in comparison with populations on other host plants, for example Citrus unshiu and Ilex crenata. Reproductive incompatibility between P. citri populations on C. unshiu and Osmanthus fragrans was found. Therefore, in Japan, P. citri differentiates into two groups one of which associates with Osmanthus, and the other with other host plants. This revised version was published online in November 2006 with corrections to the Cover Date.     

Gene Flow in the Exotic Colonizing LadybeetleHarmonia axyridisin North America

The introduced parasitoid,Aphytis melinusDeBach (Hymenoptera: Aphelinidae), is used for augmentative biological control of California red scale,Aonidiella aurantii(Maskell) (Homoptera: Diaspididae). Commercially reared wasps are reared on oleander scale,Aspidiotus neriiBouché (Homoptera: Diaspididae). Oleander scale covers lack the chemical,O-caffeoyltyrosine, a kairomone mediating host selection byA. melinus.Wasps reared on oleander scale but individually exposed, or primed, toO-caffeoyltyrosine more readily accepted California red scale for probing in laboratory bioassays and parasitized a greater proportion of available California red scale in the field than wasps reared similarly but not exposed toO-caffeoyltyrosine. Thus, it may be possible to improve host recognition of commercial, insectary-rearedA. melinusby exposing them toO-caffeoyltyrosine prior to release. The goal of this study was to develop an inexpensive but effective means of priming thousands of wasps simultaneously toO-caffeoyltyrosine. The most effective method, but potentially the most expensive, was simply to spray parasitized oleander scale on their host plant with diluteO-caffeoyltyrosine prior to wasp emergence. In additional experiments, using controlled doses of syntheticO-caffeoyltyrosine applied to scale covers, we showed that primed wasps require both a lower minimum dose ofO-caffeoyltyrosine for recognition and also respond to measuredO-caffeoyltyrosine doses more consistently than unprimed wasps. The ability to mass-prime thousands of wasps prior to release is a crucial step toward realizing the concept of behavioral improvement of host selection of parasitoids on a commercial scale.     

In Vitro Cultivation of the Vascular Phase of Sarcocystis singaporensis

To establish an in vitro culture system for the precystic phase of Sarcocystis singaporensis, we initially tested various excysting fluids for sporocysts. An excysting fluid containing 2.5% bovine taurocholate and 10% bile of the specific intermediate host, Rattus norvegicus, in RPMI medium was the most suitable resulting in excystation of 80% of the sporozoites. Subsequently, we identified brain endothelial cells and pneumonocytes of the rat to promote growth of sporozoites to schizonts. Hepatoma, fibroblastic, or myoblastic cells were not suitable for the parasite's development. First-generation schizonts were seen at days 3-10 postinoculation (PI); a distinct second peak of schizogonic development only occurred in endothelial cells at days 14-18 PI. First-generation schizonts were 26.0 (± 3.8) μm in diameter and contained 32-50 merozoites, second-generation schizonts measured 34.4 (± 10.6) μm and contained 54-72 merozoites. Merozoite yield at large-scale culture conditions (75 cm² flasks) using pneumonocytes as host cells was relatively low. Ultrastructurally, sporozoites and merozoites were quite similar to corresponding stages of other Sarcocystis species. With regard to host cell specificity and developmental kinetics, in vitro cultivation showed close similarities to the situation in vivo.     

Oxidatively Damaged Erythrocytes Are Recognized by Membrane Proteins of Macrophages

Human erythrocytes incubated with an iron catalyst ADP-chelated Fe³⁺ undergo oxidative damage of the membrane including lipid peroxidation, protein oxidation, and protein aggregation, and become susceptible to recognition by human macrophages. In order to clarify the membrane components of macrophages responsible for the recogrution of the oxidized erythrocytes, binding of the oxidized cells to dot and Western blots of solubilized membrane of macrophages was investigated. The oxidized erythrocytes but not unoxidized cells bound to the dot blots. The binding was effectively inhibited by saccharide chains of band 3, a major glycoprotein of human erythrocytes, and lowered when the saccharide chains of band 3 were removed from the cell surface by pretreatment of the cells with endo-P-galactosidase which specifically cleaves the polylactosaminyl saccharide chains of band 3. The oxidized erythrocytes bound to the membrane proteins of macrophages with molecular mass of about 50, 80, and 120 kDa on Western blots depending on the saccharide chains of band 3 on their surface. The results suggest that the oxidatively damaged erythrocytes are specifically recognized by these proteins of macrophage membrane having saccharide binding ability.     

αB-Crystallin Interacts with Cytoplasmic Intermediate Filament Bundles during Mitosis

The small heat-shock protein αB-crystallin interacts with intermediate filament proteins. Using cosedimentation assay, we showed previously that in vitro binding of αB-crystallin to peripherin and vimentin was temperature-dependent. Furthermore, when NIH 3T3 cells were submitted to different stress conditions a dynamic reorganization of the intermediate filament network was observed concomitantly with the recruitment of αB-crystallins on the intermediate filament proteins. Thus, the intracellular state of αB-crystallin correlated directly with the remodeling of the intermediate filament network in response to stress. Here, we show data suggesting that αB-crystallin is implicated in remodeling of intermediate filaments during cell division. We investigated the intracellular distribution of αB-crystallin in naturally occurring mitotic NIH 3T3 cells and in neuroblastoma N2a and N1E115 cells. In NIH 3T3 cells, αB-crystallin remained diffused throughout the cell cycle. Subcellular fractionation of αB-crystallin showed that αB-crystallin remained in the cytosolic compartment during mitosis. Furthermore, αB-crystallin accumulated in mitotically arrested NIH 3T3 cells. This increased level of αB-crystallin protein was due to an increased level of αB-crystallin mRNA in mitotic NIH 3T3 cells. In the neuroblastoma cells, the intermediate filaments were rearranged into thick cable-like structures and αB-crystallin was recruited onto them. In neuroblastoma N2a cells the level of expression did not change during the cell cycle. However, a small fraction of αB-crystallin switched onto the insoluble fraction in mitotically arrested N2a cells. Our results suggested that depending on the state of rearrangement of the intermediate filament network during mitosis αB-crystallin was either recruited onto the intermediate filaments or upregulated in the cytosolic compartment.