The location of parasites within their hosts: the behavioural component in the larval migration of Nippostrongylus bras1l1ensis in the tissues of the rat

The role of larval behaviour in successful completion of tissue migration is briefly discussed and it is related to the passive carriage of larvae along the ‘pipes and tubes’ of the host. Larvae of N. brasiliensis were injected into selected portions of the circulatory system and following periods of 5–60 min they were recovered from the blood, liver and lungs. Larvae were also immobilised in 0·4% piperazine, a dosage which permitted recovery in about 60 min. The dispersion of treated larvae was compared with that untreated controls. It was found that larvae were carried very rapidly in the blood stream and that they became lodged in the first capillary bed that they entered. They could not pass through capillary beds without movements (and/or secretions). A decreased number of adults developed after larvae were introduced via a series of routes which required the larvae to pass through an increasing number of ‘hurdles’ to migration.     

Short-term fluctuation in salinity promotes rapid larval development and metamorphosis in Dendraster excentricus

The effect of constant and fluctuating salinity on larval development and metamorphosis of the sand dollar Dendraster excentricus was investigated in the laboratory. Sand dollar larvae at different stages of development were kept either at 32‰ (controls), exposed to constant low salinity (22‰) throughout development, or exposed to fluctuating salinity (i.e. transferring larvae from 32‰ to 22‰ for 7 days then back to 32‰ for the rest of their development). Larvae exposed to constant low salinity were significantly smaller but developed all larval arms at a slower rate than larvae in all other treatments. Larvae exposed to fluctuating salinity recovered and developed significantly longer larval arms and bigger rudiments than larvae kept at constant low salinity. Larvae exposed to fluctuating salinity produced more juveniles than larvae at constant high salinity (32‰), while those at constant low salinity produced few or no juveniles. Four-arm larvae exposed to fluctuating salinity produced significantly more juveniles than six-arm larvae exposed to the same treatment. Transferring competent 8-arm larvae from 31‰ to 15‰ for 2 days then back to 31‰, induced metamorphosis with juvenile production being significantly higher than for those kept at a constant salinity of 20, 25 and 31‰. This study indicates that a short-term decrease in salinity might induce metamorphosis for this species.     

Characteristics of larvae of the southwestern corn borer, Diatraea grandiosella, obtained from populations present in tropical and temperate regions

A comparative study was undertaken of southwestern corn borers (Diatraea grandiosella) collected from south central Mexico (19°N latitude) and southeast Missouri (37°N latitude). All the life stages of the Mexican insects were found to be larger or heavier, or both, than were those of the Missouri insects. Mexican larvae grew at a higher rate and attained a significantly heavier body weight than did Missouri larvae. Although both Mexican and Missouri larvae underwent ecdyses during diapause, Mexican larvae ecdysed more frequently than did the Missouri larvae at 23°C light:dark 12 h:12 h. Larvae that ecdysed most frequently did not necessarily spend longer in diapause. Electrophoresis of the fat body and haemolymph proteins of Mexican and Missouri larvae revealed similar patterns. The fat body of diapausing Mexican larvae contained substantial amounts of a diapause-associated protein which has been characterized previously from the fat body of Missouri Larvae. Double immunodiffusion confirmed that the diapause-associated protein of the Mexican larvae was identical to that present in the Missouri larvae. Smaller amounts of this protein appear to be present in the fat body of diapausing Mexican larvae than are present in that of diapausing Missouri larvae.     

In vitro migration of Dictyocaulus viviparus larvae: migration of the infective stage inagar gel.

The infective stage of the cattle lungworm, Dictyocaulus viviparus, was able to migrate in agar gel when activated by bile. The number of larvae which penetrated the upper surface of a 2 mm thick agar layer was counted and found to be independent from the agar concentration. Larvae which had migrated out of the agar remained on the surface and did not re-enter. The agar migration process was temperature dependent. Influence of time as well as dependency of agar thickness was investigated. The significance of these findings is discussed.     

Factors Influencing Survival of Ditylenchus dipsaci (Kühn, 1857) in Soil

Ditylenchus dipsaci larvae survived in soil without a host plant for at least 242 days when held at 15 C and 21 C. Larvae held at 15 C remained infective for 212 days. Moisture levels within both clayey and sandy soils did not appreciably affect recovery of larvae. Active nematodes recovered from soil are not necessarily infective. Temperatures of -12, 0 and 4 C had little adverse effect on larvae in infected leaf tissues in soil. Larvae in soil exposed to 0 C for short periods of time were not affected adversely. Recovery of larvae from sandy soil by Baermann funnels was significantly better at 24 C than at 4 C. Differences in recovery from clay soil were not significant at these temperatures.     

Studies on the group effect in Acheta domesticus (L.) using artificial diets

Larvae of Acheta domesticus were reared on artificial diets both singly and in groups of six. The growth of group-reared larvae was significantly faster, and survival of singly reared larvae was greatly reduced. Survival of singly reared larvae was raised to that of grouped larvae when 1% wheat-germ oil was added to the diet. Wheat-germ oil also improved the growth of grouped larvae and of singly reared female larvae. The fatty acid composition of body lipid was similar, except for linolenic acid, whether the larvae were singly or group-reared, or whether wheat-germ oil was fed or not. These results are discussed from the point of view of the mechanism of the group effect.     

Environmentally Controlled Sex Expression in Meloidodera floridensis

Larvae of Meloidodera floridensis develop as females after feeding on pine roots, but become males under conditions of starvation. Seventy to 80% of the larvae kept in tap water at 23 C for 4 months underwent one or two molts, developing as males, and more than 50% became adult males. Ninety-six percent of the larvae that entered pine roots became females and only 4% developed as males. There is evidence that the latter did not feed on the roots. In comparison with tap water, solutions of cholesterol, testosterone propionate and β-estradiol did not significantly affect the percentage of larvae that developed into males. Larvae kept in soil without a host plant did not develop into males. Most of them exhausted their energy supply and died without undergoing any development. We conclude that sex expression in M. floridensis is to a large extent controlled by environmental factors. Under natural conditions of feeding on a host plant, larvae develop as females according to their genetic constitution (thelytokous organism). Under conditions of starvation, however, sexual differentiation proceeds toward the male direction, probably as a result of alteration of the hormonal balance of the larvae and the subsequent activation of different sites of genetic function.     

Susceptibility of early larval stages of Pseudaletia unipuncta and Spodoptera exigua (Lepidoptera: Noctuidae) to the entomogenous nematode Steinernema feltiae (Rhabditida: Steinernematidae)

Early stages (neonate to 7- or 8-day-old larvae) of Spodoptera exigua and Pseudaletia unipuncta were exposed to the entomogenous nematode, Steinernema feltiae, at concentrations of 0, 10, 25, 60, 100, or 200 nematodes per larva. Larvae of both species were susceptible to nematode infections. However, neonate larvae of S. exigua were significantly less susceptible to nematode infection than 3- or 8-day-old larvae at or above 50 nematodes per larva. Mortalities of neonate larvae exposed to 50 or more nematodes ranged from 68 to 74% while mortalities of 3- and 8-day-old larvae ranged from 91 to 100%. The results with P. unipuncta showed similar trends as described for S. exigua, albeit at a lower mortality level and usually with no statistical differences. Mortalities of neonate larvae exposed to 50 or more nematodes ranged from 34 to 44% while mortalities of 7-day-old larvae ranged from 32 to 91%.     

The location of parasites within their hosts: The passage of Nippostrongylus brasiliensis through the lungs of the laboratory rat

This paper examines those factors which determine the emergence of N. brasiliensis from the pulmonary circulation into the alveoli and bronchioles of the rat's lung. After injection into the superior vena cava, larvae were found to be distributed throughout the lobes of the lungs in direct relation to the mass of blood and tissue and it was concluded that this resulted from entirely random forces. Larvae which had been reversibly immobilized in piperazine were trapped in the capillaries of the lungs in the same way as untreated larvae; this accumulation was considered to be passive. Larvae could penetrate surgically collapsed lungs but did so in lower numbers than in functional lungs so host breathing movements may facilitate penetration but are not essential. Larvae could repeat their migrations in a succession of hosts, but only if removed from the lungs within 2 h, by 20 h they could not and were destroyed in the lung tissue. Larvae were passively trapped in the arterioles but can emerge only through their own behavioural efforts. Heavy larval doses (10,000) caused considerable reversible oedema and haemorrhage. This was due to the behaviour of the larvae and not the direct effect of mechanical blockage because the same was not true if 10,000 heat-killed larvae were injected. Larvae of Ancylostomatubae forme, which does not secrete enzymes, also caused haemorrhaging in rat lungs, demonstrating that larval motility alone can lead to emergence through rupturing arterioles. By 20 h larvae were active in the trachea but they showed no directional orientation and were carried passively in the mucus. At this stage they were strongly photonegative.     

Disruptive developmental effects of benzyl-1,3-benzodioxole derivatives on unparasitized and parasitized Manduca sexta larvae

Treatment of tobacco hornworm larvae with the benzyl-1,3-benzodioxole derivative J-2710 immediately after ecdysis to the fourth instar disrupted development either during the moult to the fifth instar or shortly thereafter. Larvae given topical applications of 100 μg J-2710 in 1 μl acetone suffered 100% mortality, often after secreting moulting fluid in large pockets between the epidermis and the cuticle later in the fourth instar. Larvae that successfully ecdysed had abnormalities of the mouthparts and cervix that interfered with normal feeding, inhibiting growth in the fifth instar. Larvae of the gregarious endoparasitic wasp Cotesia congregata (=Apanteles congregatus) frequently failed to emerge from host Manduca sexta larvae treated with high doses of J-2710, particularly when the host failed to feed normally. Less potent disruptive effects on Manduca and Cotesia were seen after treatment of larvae with the derivatives J-3370 and J-2581.No anti-juvenile hormone action of J-2710 was observed. J-2710-treated M. sexta larvae showed no precocious metamorphosis and the developmental effects of J-2710 were not prevented by co-application of the juvenile hormone analogue methoprene in doses ranging from 1 to 100 μg/larva. Moreover, J-2710 had no effect on the action of methoprene in the black larval assay for juvenile hormone-like activity, unlike results reported to occur using the Galleria wax wound assay.     

The effects of DOPA decarboxylase inhibitors on the permeability and ultrastructure of the larval cuticle of the Australian sheep blowfly, Lucilia cuprina

Larvae of Lucilia cuprina, fed toxic levels of α-methyl DOPA (or other DOPA decarboxylase inhibitors) during the first or second instar, die at the completion of the next moult, soon after exposing their new cuticles. In electron micrographs of newly synthesised cuticle from these treated larvae, the ultrastructure of the lipid-rich outer epicuticle layer appears to be abnormal. This newly formed cuticle of the treated larvae is apparently defective in its role as a water permeability barrier (compared with that of normal larvae), since it permits the free movement of water in both directions. Thus, treated larvae die most probably as a direct result of dehydration. Larvae fed toxic levels of α-methyl DOPA can be rescued from death by simultaneously adding N-acetyldopamine (the cuticular sclerotizing agent) to the food. The rescued larvae are apparently normal in all respects. This suggests that sclerotization is required for the formation of a normal outer epicuticle. Diflubenzuron, which is known to inhibit chitin deposition in the cuticles of a number of different species of insect, also apparently affects chitin deposition in the larval cuticle of L. cuprina. Thus, in electron micrographs of cuticle from larvae fed toxic levels of diflubenzuron the ultrastructure of the chitin-containing endocuticle layer appears to be abnormal.     

Feeding behavior of Simulium vittatum larvae in response to various suspended materials: implications for control using biological larvicides

Biologically based larvicides used to suppress nuisance populations of black flies (Diptera: Simuliidae) require that the larvae ingest the larvicide for mortality to occur. Materials present in the habitat that alter larval feeding behavior may reduce the effectiveness of the larvicide. This study was conducted to determine the effects that commonly occurring materials in larval habitats have on Simulium vittatum Zetterstedt feeding behavior. Larvae were exposed to 50 p.p.m. of kaolinite clay and 4 p.p.m. of purified diatom frustules. Larvae were also exposed to 10 000 cells or colonies per ml of viable green algae, Chlorella vulgaris Pratt and Scenedesmus quadricauda (Turp) Bréb, and the diatom Cyclotella meneghiniana Kützing to determine the effects that these materials have on larval feeding behavior. Individual larvae were recorded using a digital camcorder to determine the average number of times the larvae extended and retracted their cephalic fans per min (flick rate) in each medium. Following exposure to the selected material, neon red particles (NRP) (DayGlo) were added to the larval medium for 20 min. A spectrophotometric analysis was conducted to determine the average quantity of NRP consumed by larvae per min in each medium. The flick rate and quantity of NRP consumed by larvae was not significantly affected by clay material or viable Chlorella cells. When Scenedesmus colonies or Cyclotella cells were present in the larval medium, larval flick rate and NRP ingestion was significantly reduced. The presence of diatom frustules in the medium resulted in the most severe reduction in larval flick rate and NRP ingestion. The results suggest that larval flick rate can change depending on environmental conditions, and that suppression programs using biologically based larvicides may benefit from investigating the potential correlation between algal counts and treatment efficacy.     

Response of Galleria mellonella (Lepidoptera: Pyralidae) and Tenebrio molitor (Coleoptera: Tenebrionidae) to Spiroplasma citri inoculation

Injections into 4th instar larvae of Galleria mellonella using either in vitro or in vivo inoculum of the BR-6 isolate of Spiroplasma citri, propagated for one to nine passages, caused 5.7 to 24.7% mortality. Weight gain of the larvae injected at their 4th, 5th, and 6th instar was reduced in the first 4 days after inoculation but final pupal weight of the survivors was not significantly affected. Fourth instar larvae pupated within 10 days after the injection, but more larvae (6–13%) injected with 5th- to 9th-passage cultures pupated 5 or more days later than did larvae injected with 1st to 4th-passage cultures (0–3%). As many as one-third of the injected larvae developed into deformed pupae, with some external appendages missing or with a reduced and distorted thorax or abdomen with uneven tanning of the integument. Spiroplasma multiplied to dense concentrations (10⁸ to 10⁹/ml) in hemolymph smears from injected larvae incubated under oil. Larvae of Tenebrio molitor were not susceptible to S. citri by injection or feeding and G. mellonella were not susceptible by feeding. Transmissibility of S. citri by leafhopper vector to celery and periwinkle plants was retained after propagation for nine successive passages during 7 months in a nonhost insect such as Galleria.     

Pelagic larvae of bivalve mollusks of Amursky Bay,Sea of Japan

Larvae of 63 Bivalvia taxa were found in Amursky Bay. Of these, 47 were identified to the species level. Larvae of 55 taxa were found throughout the bay. The highest species diversity was recorded in the northeastern part and in the center of the southern part of the bay, as well as at the exit from it. Larvae of Mactra chinensis, Mytilus coruscus, Crassostrea gigas, Teredo navalis, Mya japonica, and Mysella ventricosa were widely distributed in the bay. The maximum numbers of species were observed in July and September. A comparison of species lists using the set-theory method showed that the species complexes typical of different parts of Amursky Bay are formed under the influence of the Razdolnaya River runoff flow from the north and marine waters coming from the south. The mean density of larvae during the observation period was 1100 specimens/m³ in the northern and 735 specimens/m³ in the southern part of the bay. The highest density of larvae in Amursky Bay was observed at the end of June, the beginning of July, and in September; the minimal larval density occurred in April, May, and August. While the species diversity was similarly high, the density of larvae was significantly higher in Amursky Bay than in the nearby Ussuriisky Bay.     

Galleria mellonella Infection Model Demonstrates High Lethality of ST69 and ST127 Uropathogenic E. coli

Galleria mellonella larvae are an alternative in vivo model for investigating bacterial pathogenicity. Here, we examined the pathogenicity of 71 isolates from five leading uropathogenic E. coli (UPEC) lineages using G. mellonella larvae. Larvae were challenged with a range of inoculum doses to determine the 50% lethal dose (LD₅₀) and for analysis of survival outcome using Kaplan-Meier plots. Virulence was correlated with carriage of a panel of 29 virulence factors (VF). Larvae inoculated with ST69 and ST127 isolates (10⁴ colony-forming units/larvae) showed significantly higher mortality rates than those infected with ST73, ST95 and ST131 isolates, killing 50% of the larvae within 24 hours. Interestingly, ST131 isolates were the least virulent. We observed that ST127 isolates are significantly associated with a higher VF-score than isolates of all other STs tested (P≤0.0001), including ST69 (P<0.02), but one ST127 isolate (strain EC18) was avirulent. Comparative genomic analyses with virulent ST127 strains revealed an IS1 mediated deletion in the O-antigen cluster in strain EC18, which is likely to explain the lack of virulence in the larvae infection model. Virulence in the larvae was not correlated with serotype or phylogenetic group. This study illustrates that G. mellonella are an excellent tool for investigation of the virulence of UPEC strains. The findings also support our suggestion that the incidence of ST127 strains should be monitored, as these isolates have not yet been widely reported, but they clearly have a pathogenic potential greater than that of more widely recognised clones, including ST73, ST95 or ST131.     

Interactions between a braconid,Microplitis croceipes,and a fungus,Nomuraea rileyi,in laboratory-reared bollworm larvae

The parasite Microplitis croceipes required 1.1 days longer at 26°C to complete development in Heliothis zea larvae than was required for the fungus Nomuraea rileyi to kill the host larvae and sporulate. Host larvae parasitized by M. croceipes or infected with N. rileyi failed to complete a fifth larval molt or pupate. Of the remaining healthy larvae, one-half completed six larval stadia before popation. Larvae parasitized by M. croceipes were predisposed to infection by N. rileyi, but the fungus inhibited development of M. croceipes if host larvae were infected with N. rileyi within 1 day after parasitization.     

Host-parasite relationship of Angiostrongylus cantonensis in Lymnaea palustris. II. Histopathology.

Larvae of Angiostrongylus cantonensis cause histopathologic changes in Lymnaea palustris, being characterized by mechanical damage to cells and nonspecific cellular responses to the larvae. Amoebocytes, fibroblasts, and pigment cells accumulate around larvae, encapsulating them in nodules. The intensity of the response is variable.     

Oral exudate as a mediator of behavior in larval eastern and western spruce budworms (Lepidoptera: Tortricidae)

The production of oral exudate by larval eastern and western spruce budworms,Choristoneura fumiferana (Clem.) andChoristoneura occidentalis Free., respectively, was investigated in the laboratory. All larvae except those entering into a molt exhibited aggressive behavior and produced exudate in response to handling or intraspecific encounters. Larvae could be induced to produce exudate up to four times over 2–3 min and produced an average of 1.92±0.04 µl (X ± SE) per induction. Larvae on foliage spent much of their time maintaining their silken feeding tunnel, including spinning and combing silk and removing frass. Exposure to conspecific oral exudate deposited inside the tunnel, or released by agitated larvae inside the tunnel, increased the proportion of larvae that dispersed away from the tunnels and, apparently, increased the larval sensitivity to disturbances. The behavior induced by the oral exudate indicates that it acts as an epideictic (spacing) pheromone.     

Longevity of Toxocara cati Larvae and Pathology in Tissues of Experimentally Infected Chickens

This study was conducted to determine the distribution patterns and duration of stay of Toxocara cati larvae in organs of chickens and to investigate chronic phase and potential zoonotic risk of toxocariasis in chickens. Chickens were orally infected with 1,000 embryonated T. cati eggs and necropsied 240 days post-infection. Organs of the chickens were examined at gross and microscopic levels; tissues were digested to recover larvae. Peribronchiolitis with infiltration of lymphocytes, and hyperplasia of bronchiolar associated lymphatic tissues (BALT) and goblet cells, were evident in the lungs of infected chickens. There were mild hemorrhages and infiltration of lymphocytes and a few eosinophils in the meninges. Larvae were recovered from 30% of the exposed chickens. Larvae recovery indicated that T. cati larvae stay alive for at least 240 days in the chicken brain. Therefore, chickens may potentially act as a paratenic host in nature and transfer T. cati larvae to other hosts.     

The larva of Eustra (Coleoptera, Paussinae, Ozaenini):a facultative associate of ants

Larvae of the ground beetle genus Eustra Schmidt-Goebel are described and illustrated for the first time and some biological notes are reported. One specimen of an unknown Eustra species was collected while excavating a nest of the ant Pachycondyla javana Mayr, in Taiwan, which is the first report of a paussine associated with a member of the ant subfamily Ponerinae. Several larvae and adults of a second species, Eustra chinensis Bänninger, were collected in Shanghai under bark with no association with ants. First instar larvae of the latter species were also reared in the lab. The occurrence of larvae of the genus Eustra both inside and outside ant nests, together with a report of adults collected inside a nest in Taiwan, suggests that members of this genus may be facultative predators or facultative symbionts of ants, an attribute that has never been reported for this genus. The larvae of Eustra show several unique features, including a peculiar bidentate mandibular apex, an extremely long galea, one of two tarsal claws greatly reduced, abdominal setae (including those of terminal disk) elongate and clavate at apex, urogomphi wide and flattened, and inflated sensilla S-I. Larvae were studied by both optical and scanning electron microscopy, their morphological features are compared with those of other described Paussinae larvae, and their potential phylogenetic and functional significance are discussed.