Growth and survival of blowfly Lucilia sericata larvae under simulated wound conditions: implications for maggot debridement therapy

Maggot debridement therapy has become a well‐established method of wound debridement. Despite its success, little information is available about the optimum duration of the treatment cycle and larval growth in wounds. This study examines the development of Lucilia sericata (Diptera: Calliphoridae) larvae under two containment conditions (bagged and free range) under simulated wound conditions and assesses the impact of transport and further storage of larvae on their survival and growth. There was no significant difference in size between bagged and free‐range larvae over the 72‐h experimental period. Larvae grew fastest 8–24 h after inoculation and completed their growth at 40–48 h. Mortality rates were similar (0.12–0.23% per hour) in both containment conditions and did not differ significantly (P = 0.3212). Survival of free‐range larvae was on average 16% lower than survival of bagged larvae. Refrigeration of larvae upon simulated delivery for > 1 day reduced their survival to < 50% and caused a reduction in growth of up to 30% at 12 h, but not at 48 h, of incubation. Therefore, it is recommended that free‐range larvae are left in the wound for a maximum of 40–48 h, and bagged larvae for 48–72 h. Larvae should be used within 24 h of delivery to avoid high mortality caused by prolonged refrigeration.     

Swimming Behaviour of <Emphasis Type="Italic">Chironomus acerbiphilus</Emphasis> Larvae in Lake Katanuma

We conducted a seasonal survey of the swimming behaviour of Chironomus acerbiphilus larvae in volcanic Lake Katanuma from April 1998 to December 2001. Swimming C. acerbiphilus density was much higher than other chironomid species in lakes. All C. acerbiphilus larvae (1st through 4th instars) swam, but the earlier instars (especially the 1st) had the greatest densities and fluctuations. First instars were never found in the benthic population. This result indicates that the 1st-instar larvae are planktonic. Low water temperature (below about 10 °C) resulted in the seasonal disappearance of swimming chironomid larvae. Chemical factors – oxygen depletion or presence of hydrogen sulfide – also restricted the distribution of swimming and benthic larvae. Larvae were distributed only in the oxygen-rich part of the lake bottom and swam only in the oxygen-rich layer of the water column. The density of older swimming C. acerbiphilus (3rd and 4th instars) tended to increase with increasing benthic larval densities. The chemical stress of oxygen depletion or presence of hydrogen sulfide during holomixis within and after the stratification period leads to conspicuous swimming behaviour of benthic C. acerbiphilus larvae. Almost all C. acerbiphilus larvae died on this occasion.     

Movement of cynomolgus and rhesus monkey spermatozoa collected from the lower female reproductive tract

Postcoital (pc) cervical mucus was collected in 73 menstrual cycles of cynomolgus monkeys and in 43 cycles of rhesus monkeys at 2,6,10,30 hr pc. Videomicrography was used to analyze sperm numbers and movement in the mucus. Both cynomolgus and rhesus monkeys had comparable populations of motile sperm in the mucus at 2 hr pc. However, by 6 hr pc, cervical mucus from cynomolgus monkeys contained twice as many total sperm and motile sperm as mucus from rhesus monkeys (P <.05). Mean swimming speeds of the free-swimming cervical sperm were similar for the two species at this time. No motile sperm were recovered in mucus from rhesus monkeys at 30 hr pc. In cynomolgus monkeys, however, 14 of the 26 animals examined at 30 hr pc had motile sperm in their mucus. These sperm exhibited lower percent molility, percent free-swimming sperm, and swimming speed than those sperm observed at 6 hr pc. Uterine sperm were collected by transcervical or transuterine aspiration from cynomolgus monkeys. In the transcervical technique, sperm were successfully obtained in four of nine animals examined at 6 hr and in four of five animals at 30 hr pc. The percentage of motile sperm in the uterine fluid was high, 82% ± 4%, and the swimming speeds (86 ± 2μm/sec) were higher than those observed in cervical mucus. Approximately 5–10% of the uterine sperm exhibited swimming motions similar to the hyperactivated motility seen in most mammals. These findings indicate that the sperm cervical mucus interaction in vivo in cynomolgus monkeys has more similarities to the human situation than does the interaction in rhesus monkeys.     

APPLIED ISSUES The effect of ammonium nitrate on the feeding and development of larvae of the smooth newt, Triturus vulgaris (L.), and on the behaviour of its food source, Daphnia

1. Larvae of the smooth newt, Triturus vulgaris, were exposed to four concentrations of ammonium nitrate in artificial pond water (50, 100, 200 and 500 mg 1-^?1) under controlled laboratory conditions. 2. Larvae exposed to a 50 mg 1-^?1 solution of ammonium nitrate showed no significant difference to control larvae in feeding rate, mass at metamorphosis or time to metamorphosis. However, larvae exposed to 200 or 500 mg 1-^?1 ammonium nitrate, for a single period of 24, 48 or 72 h, were significantly smaller than controls at metamorphosis. Larvae exposed to 100 mg 1-^?1 had a significantly higher feeding rate than those reared under control conditions or those exposed to 200 or 500 mg 1-^?1 ammonium nitrate, but this was not reflected in their size at metamorphosis, which was not significantly different from the controls. Larval survival remained high in all trials. 3. In separate trials, Daphnia, used as a food source for the newt larvae, were exposed to the same series of ammonium nitrate concentrations. Daphnia behaviour was affected by exposure to 200 and 500 mg 1-^?1 ammonium nitrate, and in both cases animals were significantly more likely to be found at the top of the water column than those exposed to 50 or 100 mg 1-^?1 ammonium nitrate or the control medium.     

Three‐dimensional fate mapping of larval tissues through metamorphosis in the glass sponge Oopsacas minuta

The tissue of glass sponges (Class Hexactinellida) is unique among metazoans in being largely syncytial, a state that arises during early embryogenesis when blastomeres fuse. In addition, hexactinellids are one of only two poriferan groups that already have clearly formed flagellated chambers as larvae. The fate of the larval chambers and of other tissues during metamorphosis is unknown. One species of hexactinellid, Oopsacas minuta, is found in submarine caves in the Mediterranean and is reproductive year round, which facilitates developmental studies; however, describing metamorphosis has been a challenge because the syncytial nature of the tissue makes it difficult to trace the fates using conventional cell tracking markers. We used three‐dimensional models to map the fate of larval tissues of O. minuta through metamorphosis and provide the first detailed account of larval tissue reorganization at metamorphosis of a glass sponge larva. Larvae settle on their anterior swimming pole or on one side. The multiciliated cells that formed a belt around the larva are discarded during the first stage of metamorphosis. We found that larval flagellated chambers are retained throughout metamorphosis and become the kernels of the first pumping chambers of the juvenile sponge. As larvae of O. minuta settle, larval chambers are enlarged by syncytial tissues containing yolk inclusions. Lipid inclusions at the basal attachment site gradually became smaller during the six weeks of our study. In O. minuta, the flagellated chambers that differentiate in the larva become the post‐metamorphic flagellated chambers, which corroborate the view that internalization of these chambers during embryogenesis is a process that resembles gastrulation processes in other animals.     

Settlement behaviour of larvae of the Stripey Snapper, <Emphasis Type="Italic">Lutjanus carponotatus</Emphasis> (Teleostei: Lutjanidae)

Larval behaviour is important to dispersal and settlement, but is seldom quantified. Behavioural capabilities of larval Lutjanus carponotatus in both offshore pelagic and reef environments at Lizard Island, Great Barrier Reef were observed in situ to determine if they were sufficient to influence dispersal. Offshore, larvae swam with higher directional precision and faster on the windward side of the island (28 cm.s⁻¹) than on the leeward side (16 cm s⁻¹). Most larvae swam directionally. Mean swimming directions were southerly in the windward area and northerly in the leeward area. Larvae avoided the surface and remained mostly between 3–15 m. Larvae released near reefs were 2–3 times faster swimming away from reefs (19 cm s⁻¹) than swimming toward or over them (6–8 cm s⁻¹). Speed swimming away was similar to that offshore. Of 41 larvae released near reefs, 73% reached the reef, 59% settled, and 13% of those reaching the reef were eaten. Larvae settled onto hard and soft coral (58%), topographic reef features (29%) and sand and rubble (13%). Settlement depth averaged 5.5 m (2–8 m). Before settling larvae spent up to 800 s over the reef (mean 231 s) and swam up to 53 m (mean 14 m). About half of the larvae interacted with reef residents including predatory attacks and aggressive approaches by residents and aggressive approaches by settling larvae. Settlement behaviour of L. carponotatus was more similar to a serranid than to pomacentrids. Settlement-stage larvae of L. carponotatus are behaviourally capable, and have a complex settlement behaviour.     

Efficacy of entomopathogenic nematode,Steinernema carpocapsae against the diamondback moth,Plutella xylostella (L.) in laboratory condition

In vitro studies were carried out on the diamondback moth, Plutella xylostella larvae using an insect entomopathogenic nematode isolate, Steinernema carpocapsae obtained from the Koppert company, the Netherlands. Larvae of P. xylostella were collected from cabbage farms around Mashhad city of Iran. During the study, the responses of larvae at 25?°C for three periods of 24, 48 and 72?h with different concentrations of 0, 5, 10, 20, 40, 80, 160 and 320 third instar larvae of nematode (infective stage?=?IJs) per insect into 10?cm Petri dishes containing filter paper soaked with 1?ml of nematodes suspension were compared. Maximum mortality caused by S. carpocapsae nematode was 88% at 24?h, and it was 100% at 48 and 72 h. With increasing nematode population level and exposure time (ET in hour), mortality of P. xylostella larvae was increased. Based on probit analysis, LC₅₀ values of S. carpocapsae nematode in three test periods were 45.61, 12.02 and 40.80 IJs per insect, respectively. Initial ANOVA was performed for S. carpocapsae nematode. The effect of both nematode population levels (IJ) and ET on third instar larvae of the diamondback moth, P. xylostella and interaction between IJ and ET were significant. In general, it is recommended to apply this nematode in suitable condition for controlling diamondback moth.     

Morphological effects of insect growth regulating compounds on Aedes aegypti (Diptera: Culicidae) larvae.

Cuticular development of $\text{Aedes aegypti}$ larvae was examined by electron microscopy and comparisons were made between larvae exposed to methoprene, isopropyl (E, E)-11-methoxy-3, 7, 11-trimethyl-2, 4-dodecadienoate, those treated with the fluorescent insect growth regulator, 5-[[[5-(dimethylamino)-1-naphthalenyl]-sulfonyl]amino]-1, 3-benzodioxole (DNSAB), and untreated larvae. Larvae of all three groups were routinely fixed at 24, 48, and 72 hr posttreatment. Thin sections of the sixth-abdominal segment, anal papillae, midgut tissue, and Malpighian tubules were examined for morphological variations from controls.     

Factors affecting oral regurgitation by larval spruce budworm

This study evaluated factors that influence the regurgitation behaviour of sixth instar spruce budworm, Choristoneura fumiferana Clemens (Lepidoptera: Tortricidae), reared on balsam fir, Abies balsamea (L.) Mill. (Pinaceae), under various experimental conditions in the laboratory. Upon physical disturbance, larvae discharged a median volume of regurgitant of 0.4 μl when fed and 1.6 μl when food‐deprived. Larvae deprived of food for 24 or 48 h disgorged more regurgitant than larvae feeding on balsam fir foliage, and the effect was consistent for laboratory‐reared and field‐collected larvae. The water content of the foliage fed upon by larvae had no immediate impact on the volume of regurgitant; following a 24‐h period of food deprivation, however, larvae that previously fed on fresh foliage discharged >2.5 times more regurgitant than larvae that previously fed on dry foliage. Self‐regulated regurgitation by larvae, measured using the number of regurgitant stains on filter paper, was >10 times higher when larvae had access to balsam fir foliage than when they were starved. The number of larvae confined inside the Petri dish (one or four individuals) had a relatively small effect on regurgitation. Larvae were deterred from feeding when balsam fir needles were entirely covered with regurgitant, but not when only a portion of the foliage was treated. These results suggest that the regurgitant does not serve as resource marking or spacing pheromone. The high level of regurgitation by larvae after contact with ants suggests that the regurgitant has evolved in part as a defence mechanism against natural enemies.     

Influence of Predator Presence and Prey Density on Behavior and Growth of Damselfly Larvae (Ischnura elegans) (Odonata: Zygoptera)

Foraging behavior is often determined by the conflicting benefits of energy gain and the risk of mortality from predation or other causes. Theory predicts that animals should have lower activity levels when either the risk of predation or the availability of resources in the environment is high. We investigated the adjustment of the behavior of I. elegans larvae to predator presence (Anax imperator) and prey density (Daphnia sp.) and their interaction in a completely crossed factorial experiment in the lab and the effect of behavior on growth. The foraging activity of the I. elegans larvae was significantly reduced in the presence of a free-swimming predator but not a caged predator. Abdominal movements were significantly reduced at a low prey density. Growth was significantly reduced by the presence of a free swimming predator and low prey densities. These results provide evidence that these damselfly larvae adjust their behavior to the presence of predators to increase their survival at the expense of reduced growth and development.     

Spatiotemporal occurrence and feeding habits of tonguefish,Cynoglossus lighti Norman, 1925, larvae in Ariake Bay,Japan

Spatiotemporal occurrence and feeding habits of tonguefish (Cynoglossus lighti Norman, 1925) larvae were investigated in an offshore area (>5 m in depth) of the inner part of Ariake Bay, Japan. All specimens were symmetric, free‐swimming larvae. Although their seasonal abundance and distribution in the study site varied from year to year, spawning started in June and the larval abundance was high in August and September with a wide distribution in the inner part of the bay. Both present and previous study results strongly suggest that larvae may settle primarily in the estuary and near‐shore areas of Ariake Bay after their wide distribution in the offshore area of the bay during the free‐swimming stage. Larvae showed a clear feeding rhythm in which they fed on prey mainly during the daytime. Larvae fed exclusively on copepods, and identified prey were mostly Paracalanidae (mainly Parvocalanus crassirostris), Microsetella norvegica, and Oithona davisae. Pre‐metamorphosis larvae fed primarily on Paracalanidae and O. davisae, whereas O. davisae formed a smaller proportion of the early‐metamorphosis diet. In early metamorphoses, larvae fed preferentially on Paracalanidae and M. norvegica.     

Aggressive Interactions and Risk of Fish Predation for Larval Damselflies

Larval damselflies frequently engage in aggressive interactions that may increase their risk of fish predation. To test this we analyzed the behavior of larval Ischnura verticalis exposed to both conspecifics and fish predators. Larvae in the presence of conspecifics oriented, struck, and swam more but crawled less compared to solitary larvae; the presence of fish reduced, or tended to reduce, all behaviors. Fish struck more at interacting larvae compared to noninteracting larvae. Increased attack rate by fish likely reflects the increase in the very active swimming behavior by larvae and suggests a conflict between antipredator behaviors. Swimming is an appropriate response to avoid predation by odonate larvae which normally ambush prey but is clearly dangerous when fast-swimming fish that cue in on movement are nearby.     

THE CYTOLOGICAL EFFECT OF ECDYSTERONE ON THE MIDGUT CELLS OF THE FLESH-FLY SARCOPHAGA BULLATA

Larvae of the flesh-fly, Sarcophaga bullata, were injected with the synthetic moulting hormone ecdysterone or saline at the beginning of the third and final larval instar. One group was left untreated. The ecdysterone-injected larvae showed an increase in number of secondary lysosomes in the midgut epithelial cells similar to that observed at the onset of metamorphosis, an event which would normally occur about 48 hr later in these larvae.     

Can bivalve veligers escape feeding currents of adult bivalves?

While the stock of introduced Pacific oysters (Crassostrea gigas) increased in the Oosterschelde estuary (SW Netherlands), so did the filtration pressure of all bivalve species together. In the same period, stocks of native bivalves declined slightly. The expansion of Pacific oysters in Dutch estuaries might be partially due to better abilities of their larvae to avoid or escape filtration, compared to larvae of native bivalves. In this context, escape and swimming abilities of Pacific oyster larvae and the larvae of the native blue mussel (Mytilus edulis) were compared.Swimming behaviour of C. gigas larvae and larvae of M. edulis was recorded in still water and in a suction current mimicking a bivalve feeding current, in a horizontal and in a vertical plane. Larval swimming behaviour in a suction flow field was reconstructed by subtracting local water movement vectors from the total movement of larvae, yielding movement paths due to larval swimming alone.Swimming speeds and the rate of displacement in vertical direction of C. gigas and M. edulis larvae were related to larval shell length, and to the pitch of up- or downward swimming.Larvae of both species did not show escape reactions in a suction flow field. With increasing shell length, larval swimming speeds of both species increased significantly. Swimming speeds of C. gigas larvae were significantly higher than swimming speeds of M. edulis larvae, resulting in a faster vertical displacement. The ability to migrate to more favourable water layers faster may offer C. gigas an advantage over native bivalves with slower swimming larvae.     

Influence of low temperature on pupation induction in Costelytra zealandica (Coleoptera: Scarabaeidae)

Costelytra zealandica (White) lives within the soil throughout its life cycle, except for brief, crepuscular excursions to the surface by the adults to mate and feed. Larvae collected more than 18 weeks before 50% field pupation did not pupate unless they were artificially chilled. Such chilling had no significant effect on the percentage pupation of larvae collected after this time; it is assumed that these larvae had received sufficient chilling from their environment to induce pupation. Larger larvae were more likely to pupate than smaller larvae during the period 13–7 weeks before 50% field pupation. It is postulated that larvae remain quiescent until the soil temperature increases in the spring, and pupate only then. Such a process would explain how the adults of a population emerge at about the same time.     

SEQUENTIAL REPRODUCTION BY DIFFERENT TYPES OF SPECIMENS OF THE ESTUARINE SPONGE,HALICHONDRIA SP., WITH AN EMPHASIS ON REPRODUCTION OF POSTLARVAL SPECIMENS

Young specimens of Halichondria sp., recently developed from larvae (postlarval specimens), engage in sexual reproduction; and reproductive activity increases with specimen size. Reproduction by such specimens takes place primarily during a period after breeding by the parental generation has declined. This fact suggests that environmental conditions are favorable for reproduction for an extended period of time and that reproduction is to a large extent under endogenous control.     

Trypanosoma cruzi: morphogenesis in diffusion chambers in the mouse peritoneal cavity and attempted stimulation of host immunity

Sequential morphologic changes and antigen producing capacity of Trypanosoma cruzi in peritoneally implanted diffusion chambers were studied. Diffusion chambers were equipped with two Nuclepore filters (0.20 μm pore size) sandwiched between three Lucite rings. Epimastigotes or trypomastigotes and amastigotes were placed in diffusion chambers and surgically implanted into the peritoneal cavity of mice, or placed in in vitro cell culture, or in various types of culture media and incubated at 26 or 37 C.Epimastigotes maintained in diffusion chambers in mice changed into trypomastigotes as evidenced by the presence of numerous transitional stages and the concomitant decrease in the percentage of the former and increase in the percentage of the latter in chambers removed and examined at 16, 24, 36, 48, 72, and 84 hr after implantation. The maximum of 68% trypomastigotes was noted in chambers examined at 84 hr. Amastigotes subsequently appeared, apparently arising from trypomastigotes and reached the highest percentage (49%) obtained at 132 hr. The total number of parasites in chambers decreased slightly during the first 36 hr (20%). Little change in the total number of parasites was noted during the interval of 36–108 hr. A subsequent decrease in numbers of parasites was noted until by 280 hr after implantation, chambers contained less than 2% of the original number of organisms present in the chambers. No similar transformation of epimastigotes was noted in diffusion chambers maintained in cell culture at 37 C or in a cell culture growth medium or LIT medium at 37 or 26 C.No detectable morphological change was noted when trypomastigotes and amastigotes were implanted in diffusion chambers in the peritoneal cavity of mice. The total number of these parasites decreased notably (82%) after 24 hr.Mice receiving diffusion chambers containing epimastigotes implanted at two different intervals (21 days apart), developed only marginal protective immunity when challenged with virulent T. cruzi three weeks after the second implant of chambers, and no protection was afforded those mice implanted with chambers containing trypomastigotes and amastigotes. Sera collected from mice 6 wk after the second implantation of diffusion chambers containing parasites were observed to have antibody titers to T. cruzi as demonstrated by the fluorescent antibody technique and direct agglutination procedure.     

Behavioural significance of pressure responses in megalopa larvae of Callinectes sapidus and Macropipus sp.

The primarily benthic megalopa larvae of Callinectes sapidus and Macropipus holsatus swim upwards for a short distance from the bottom when subjected to pressure increased above ambient values, particularly when illuminated from above. Larvae exposed to stepwise increments of pressure exhibit a threshold in the barokinetic response which is lower in Callinectes (0.4 atm) than in Macropipus (0.8–1.0 atm). Such behaviour would tend to inhibit swimming in surface waters of stratified estuaries where net flow is seawards, but would promote slight upward swimming when on the bottom in deeper water which has a net flow inwards, thus substantiating other evidence that the megalopa is a stage at which recruitment to estuaries takes place. The lower threshold of the barokinetic response in Callinectes may be related to its occurrence in more uniformly shallow estuaries on the south and east coasts of N. America.

In both species swimming is markedly inhibited after a slight reduction in pressure. Such responses would be of value to larvae running the risk of being washed ashore in surf Sudden reductions in pressure by surf action would induce sinking into the undertow which would carry larvae offshore into deeper water.     

Female dimorphism in honey-bee larvae with respect to glucose metabolism

The rate of conversion of glucose-1-¹⁴C and glucose-6-¹⁴C to ¹⁴CO₂ and lipid was monitored in queen and worker larvae between 48 and 96 hr of age. A definite dimorphism in glucose metabolism between castes was established. Worker larvae 72 hr of age have a much greater $\text{C}{}_6\text{C}{}_1$ ratio than do queen larvae of the same age. The ratios were of the same order of magnitude for both castes of larvae younger or older than 72 hr. Queen larvae were shown to have a greater rate of lipid synthesis than worker larvae.     

Behavior and development of larvae in the sponge Haliclona amboinensis

Sponges play important roles in marine ecosystems by contributing to habitat complexity and benthopelagic coupling of nutrients. Yet, the reproduction and settlement behaviors of diverse sponge species are not well understood. Here, we examined the brooding demosponge Haliclona amboinensis, which is common on shallow reefs in Bolinao, northwestern Philippines. Gravid sponges were found between the months of May and August, coinciding with warmer sea surface temperature. Sponges released parenchymella larvae from brood chambers in the mid‐morning, suggesting that light and temperature may serve as cues to initiate hatching. Larvae immediately swam toward the surface upon emergence and migrated to the bottom of the tanks 1–2 hr after release. The presence of light and crustose coralline algae induced high larval settlement. Metamorphosis proceeded rapidly in vitro, with larval cells spreading laterally on the substrate. The osculum was first visible at 3 days after settlement. The short pelagic duration of larvae in H. amboinensis promotes local recruitment and may be important for the maintenance of sponge populations in the face of disturbances.