RESUMO
Necrophagous flies that colonize human and animal corpses are extremely efficient at locating and utilizing carrion. Adult flies deposit eggs or larvae on the ephemeral food resource, which signals the beginning of intense inter- and intra-species competition. Within a short period of time after egg hatch, large larval aggregations or maggot masses form. A period of intense larval feeding ensues that will culminate with consumption/decomposition of all soft tissues associated with the corpse. Perhaps the most distinctive feature of these feeding aggregations is heat production; that is, the capacity to generate internal heat that can exceed ambient temperatures by 30°C or more. While observations of maggot mass formation and heat generation have been described in the research literature for more than 50 years, our understanding of maggot masses, particularly the physiological ecology of the aggregations as a whole, is rudimentary. In this review, an examination of what is known about the formation of maggot masses is presented, as well as arguments for the physiological benefits and limitations of developing in feeding aggregations that, at times, can represent regions of intense competition, overcrowded conditions, or a microclimate with elevated temperatures approaching or exceeding proteotoxic stress levels.
Assuntos
Comportamento Alimentar , Sarcofagídeos/fisiologia , Animais , Comportamento Competitivo , Feminino , Ciências Forenses , Temperatura Alta , Larva/fisiologia , Masculino , Densidade Demográfica , Reprodução , Vertebrados/parasitologiaRESUMO
A venom preparation from Nasonia vitripennis, a wasp ectoparasitoid of fly pupae, was assayed for lethality in different stages of insects representing ten different orders and in cultured insect cells. In most cases, the motor activity of the injected insects remained completely normal for 1-2 days after the injection and displayed none of the symptoms of paralysis commonly reported for venoms of the Hymenoptera. A natural host, the flesh fly Sarcophaga bullata, was highly sensitive in the pupal stage (LD50 = 5.4 and 5.5 VRE/g for nondiapausing and diapausing pupae, respectively), the stage that is normally parasitized, and larvae and adults were as susceptible to the venom as the pupae. Adults of another fly host, Phaenicia sericata, were nearly as sensitive (LD50 = 6.5 VRE/g), but nonhost adult flies were more tolerant. Among the other orders tested, pupae of several species (Plodia interpunctella, Trichoplusia ni, Tenebrio molitor) were more susceptible to envenomation than larval or adult stages. In fact, the highest sensitivity observed in this study (LD50 = 0.58 VRE/g) was with pupae of the cabbage looper, T. ni, a species that is not a natural host. In contrast, the larvae (LD50 = 7.23 VRE/g) and adults (LD50 = 7.48) of T. ni were far less sensitive. Adults of Nasonia vitripennis were not sensitive to their own venom (LD50 = > 533 VRE/g), although adults of another hymenopteran, Apis mellifera, were suceptible (4.62 VRE/g). Adults of Lymantria dispar, Oncopeltus fasciatus, Aphis nerii, Euborellia annulipes, Diapheromera femorata, Blattella germanica, Periplaneta americana, and Reticulitermes flavipes demonstrated a high tolerance to Nasonia venom. When tested in vitro, the venom caused cultured Lepidoptera (TN-368) and Diptera (NIH SaPe4) cells to round up, swell, and eventually die. The LC50S were 0.0014 and 0.0010 VRE/microliters for TN-368 and SaPe4 cells, respectively. Cytotoxicity was observed within 10 min after exposure to LC99 levels of venom, with 100% cell mortality at 100 min for the NIH SaPe4 cells and 24 hr for TN-368 cells. It is possible that the venom component responsible for in vivo and in vitro activities may be different, but results from the cell culture work suggest that this method offers a promising assay for quickly screening venom samples. The high susceptibility of flies and pupae of other insects to the venom, as well as its novel (nonparalytic) action suggest that it may have considerable potential for development as a biopesticide.
Assuntos
Dípteros/efeitos dos fármacos , Venenos de Vespas/toxicidade , Animais , Células Cultivadas , Dípteros/crescimento & desenvolvimento , Dípteros/parasitologia , Avaliação Pré-Clínica de Medicamentos , Mariposas/efeitos dos fármacos , Mariposas/crescimento & desenvolvimentoRESUMO
The lethal effects of crude venom prepared from the ectoparasitic wasp Nasonia vitripennis were examined with cultured cells from six insect and two vertebrate species. Venom caused cells from Sarcophaga peregrina (NIH SaPe4), Drosophila melanogaster (CRL 1963), Trichoplusia ni (TN-368 and BTI-TN-5B1-4), Spodoptera frugiperda (SF-21AE), and Lymantria dispar (IPL-Ldfbc1) to round up, swell, and eventually die. Despite similar sensitivities and overlapping LC50 values [0.0004-0.0015 venom reservoir equivalents (VRE)/microl], profound differences were noted at the onset of cytotoxicity among the six insect cell lines: over 80% of the NIH SaPe4 and SF21AE cells were nonviable within 1 h after addition of an LC99 dose of venom, whereas the other cells required a 5-10-fold longer incubation period to produce mortality approaching 100%. In contrast, cells from the grass frog, Rana pipiens (ICR-2A), and goldfish, Carassius auratus (CAR), showed little sensitivity to the venom: six venom reservoir equivalents were needed to induce 50% mortality in ICR-2A cells [50% lethal concentration (LC50) = 0.067 VRE/microl), and 9 VRE did not yield sufficient mortality in CAR cells for us to calculate an LC50. All susceptible cells showed similar responses when incubated with wasp venom: retraction of cytoplasmic extensions (when present), blebbing of the plasma membrane, swelling of the plasma and nuclear membranes, condensation of nuclear material, and eventual cell death attributed to lysis. The rate of swelling and lysis in NIH SaPe4 and BTI-TN-5B1-4 cells exposed to venom appeared to be dependent on the diffusion potential of extracellular solutes (Na+ = choline > sucrose > or = raffinose > K+), which is consistent with a colloid-osmotic lysis mechanism of cell death. When T. ni cells were cotreated with venom and the K+ channel blocker 4-aminopyridine, cell swelling and lysis increased with increasing drug concentration. In contrast, cells from S. peregrina were protected from the effects of the venom when treated in a similar manner. Addition of certain divalent cations (Zn+2 and Ca+2) to the extracellular media 1 h postvenom incubation rescued both BTI-TN-5B1-4 and NIH SaPe4 cells, suggesting that protection was gained from closure of open pores rather than prevention of pore formation. Venom from N. vitripennis displayed no hemolytic activity toward sheep erythrocytes, supporting the view that venom intoxication is not by a nondiscriminate mechanism. A possible mode of action of the venom is discussed.
Assuntos
Permeabilidade da Membrana Celular , Venenos de Vespas/farmacologia , 4-Aminopiridina/farmacologia , Animais , Morte Celular , Linhagem Celular , Tamanho Celular , Cicloeximida/farmacologia , Dípteros/citologia , Drosophila melanogaster/citologia , Carpa Dourada , Hemólise , Insetos/citologia , Cloreto de Potássio/farmacologia , Inibidores da Síntese de Proteínas/farmacologia , Rana pipiens , Cloreto de Sódio/farmacologia , Spodoptera/citologiaRESUMO
Both NAD- and NADP-dependent glyceraldehyde-3-phosphate dehydrogenase (G3PDH) (EC 1.2.1.12) activities were detected in glucose-grown cells of Pseudomonas aeruginosa strain PAO. After growth on gluconeogenic substrates such as citrate, the activity of the NAD-G3PDH was reduced severalfold in contrast to little change for the NADP-G3PDH. The two G3PDH activities could be separated by ammonium sulphate fractionation. PAGE revealed the presence of two G3PDH isoenzymes of 140 (NADP-specific) and 315 (NAD-specific) kDa. Slight differences were observed in the thermostabilities and pH optima of the two enzymes whereas the regulation of their activities by various compounds varied strongly. The NADP-G3PDH enzyme was activated by ATP, reduced NAD, and fructose 6-phosphate. It was inhibited by fructose 1,6-diphosphate and 6-phosphogluconate. The NAD-G3PDH enzyme was inhibited by ATP, reduced NAD, and 6-phosphogluconate; it was slightly activated by reduced NADP. The possible roles of these isoenzymes in the control of hexose catabolism and gluconeogenesis in P. aeruginosa are discussed.
Assuntos
Gliceraldeído-3-Fosfato Desidrogenases/metabolismo , Isoenzimas/metabolismo , Pseudomonas aeruginosa/enzimologia , Eletroforese em Gel de PoliacrilamidaRESUMO
Supercooling points (SCPs) and low temperature survival were determined for diapausing and nondiapausing larvae of the ectoparasitoid Nasonia vitripennis. Neither nondiapausing nor diapausing larvae could survive tissue freezing. The SCP profiles were nearly identical for nondiapause-destined (-27 degrees C) and diapausing larvae (-25 degrees C), but these values were not indicative of the lower limits of tolerance in either type of larvae: larvae were killed by chilling at temperatures well above the SCP. Diapausing larvae could withstand low temperature exposures 3-8 times longer than their nondiapausing counterparts. Low temperature survival was enhanced in diapausing and nondiapausing larvae by their encasement within the puparium of the host flesh fly, SARCOPHAGA CRASSIPALPIS: the LT(50)s determined for nondiapausing and diapausing larvae enclosed by fly puparia were 2-3 times higher than values calculated for larvae removed from the puparia. Additional low temperature protection was gained through acquisition of host cryoprotectants during larval feeding: nondiapausing parasitoid larvae that fed on diapausing flesh fly pupae with high levels of glycerol were able to survive exposure to a subzero temperature 4-9 times longer than wasps reared on nondiapausing fly pupae that contained lower quantities of glycerol. Alanine may also contribute to the cold hardiness of N. vitripennis, as evidenced by the fact that larvae feeding on diapausing fly pupae both contained higher concentrations of alanine and exhibited greater cold hardiness. The results thus demonstrate that several critical features of cold hardiness in the wasp are derived from biochemical and physical attributes of the host.
RESUMO
In this study, we examined cellular immune responses in the flesh fly, Sarcophaga bullata, when parasitized by the ectoparasitoid Nasonia vitripennis. In unparasitized, young pharate adults and third instar, wandering larvae of S. bullata, four main hemocyte types were identified by light microscopy: plasmatocytes, granular cells, oenocytoids, and pro-hemocytes. Parasitism of young pharate adults had a differential effect on host hemocytes; oenocytoids and pro-hemocytes appeared to be unaltered by parasitism, whereas adhesion and spreading behavior were completely inhibited in plasmatocytes and granular cells by 60 min after oviposition. The suppression of spreading behavior in granular cells lasted the duration of parasitism. Plasmatocytes were found to decline significantly during the first hour after parasitism and this drop was attributed to cell death. Melanization and clotting of host hemolymph did not occur in parasitized flies, or the onset of both events was retarded by several hours in comparison to unparasitized pharate adults. Hemocytes from envenomated flies were altered in nearly identical fashion to that observed for natural parasitism; the total number of circulating hemocytes declined sharply by 60 min post-envenomation, the number of plasmatocytes declined but not granular cells, and the ability of plasmatocytes and granular cells to spread when cultured in vitro was abolished within 1 h. As with parasitized hosts, the decrease in plasmatocytes was due to cell death, and inhibition of spreading lasted until the host died. Isolated crude venom also blocked adhesion and spreading of these hemocyte types in vitro. Thus, it appears that maternally derived venom disrupts host immune responses almost immediately following oviposition and the inhibition is permanent. The possibility that this ectoparasite disables host defenses to afford protection to feeding larvae and adult females is discussed.