Toxicity and behavioural effects of diet-borne alkaloids on larvae of the black blowfly,Phormia regina

Larvae of the black blowfly, Phormia regina (Meigen) (Diptera: Calliphoridae) were exposed for 24 h to artificial diets that contained one of the following alkaloids: arecoline, caffeine, nicotine, quinine, sparteine or strychnine at either 1000 or 100 p.p.m. Each of the alkaloids caused reduced weight gain, relative to a control population in a no-choice bioassay and, with the exception of quinine, all alkaloids caused reduced larval weights in a choice bioassay. Larvae were unable to move away from diets containing arecoline (1000 and 100 p.p.m) and congregated away from diets containing 1000 p.p.m. quinine. Arecoline (1000 p.p.m) and both concentrations of nicotine caused significant mortality of larvae. Over a longer period (120 h), 10 and 1 p.p.m. nicotine resulted in significant numbers of larvae congregating away from a treated diet. Ten p.p.m. nicotine caused reduced weight gain over 120 h, although larvae provided with a choice were less affected. Exposure of larvae to dried residues of nicotine for 2 h did not affect subsequent development.     

Verification of the eclossion mark on the sagittal otolith of the larvae of Sardinella aurita (Pisces: Clupeidae)

The study of otolith in larvae is important to determine fish age and growth, essential parameters in the study and management of fisheries resources. In this study, the formation of the hatching mark in Sardinella aurita was verified on ichthyoplankton samples collected off southern Cubagua island, Venezuela, from May 1998 to January 1999. The embryos were kept alive using a culture system until they hatched and daily a group of 10 to 30 larvae were fixed in 95% ethanol. An image analysis system was used to measure morphometric characteristics of larvae and sagittal otoliths. Following are mean values in newly hatched larvae: otolith hatching mark distance from nucleus 4.78 m (I.C. 0.36 m, p 0.05 n = 30), increase width 1.46 m (I.C. 0.17 microm, p 0.05, n = 30) and diameter 14.28 m (IC 1.11 m, p 0.05, n = 30). The mean standard length of larvae at age 0 was 3.31 mm (I.C. 0.08 mm, p 0.05, n = 200). The identification of the hatching mark allows the exact calculation of the number of rings in larvae from the natural environment.     

Evaluation of biological and chemical insecticide mixture against Aedes aegypti larvae and adults by thermal fogging in Singapore

To improve the operational efficiency of dengue vector control in Singapore, larvicide and adulticide were applied together by thermal fog generator (Agrofog AF40). The mixture consisted of Bacillus thuringiensis var. israelensis (Vectobac 12 AS) as biological larvicide at 1.5 L/ha and pirimiphos-methyl (Actellic 50 EC) as adulticide at 100 g ai/ha, diluted 10-fold with water. Aerosol of this mixture was evaluated against the mosquito Aedes aegypti (L.) (Diptera: Culicidae) in bioassays using cages of 10 adult females exposed at heights of 0.3-2.4 m and distances of 3-12 m from the hand-held generator. Cups containing 200 mL water were treated at ground level by exposure to the aerosol application at the same distances from the generator. Subsequent larval bioassays on days 1, 7, 14, 21 and 28 post-spray involved exposing 20 larvae/cup for 48 h. Droplets had VMD 57 microm and female mosquitoes were killed by 2 s exposure to the aerosol at 3 m. We obtained 92-100% mortality of the adult mosquitoes and 100% control of larvae at 3 m distance, but only 10-13% mortality at 12 m from the fogger. In treated cups, larvae showed high mortality (92%) when exposed for 48 h even 1 month post-treatment. Results demonstrate the practical advantage of using this mixture of Vectobac 12AS and Actellic 50 EC for simultaneous control of Aedes adults and larvae, with prolonged larvicidal efficacy in treated containers.     

Bioefficacy and mode-of-action of aglaroxin A from Aglaia elaeagnoidea (syn. A. roxburghiana) against Helicoverpa armigera and Spodoptera litura

The bioefficacy of aglaroxin A from Aglaia elaeagnoidea (syn. A. roxburghiana) was assessed using the gram pod borer, Helicoverpa armigera (Hübner), and Asian armyworm, Spodoptera litura (Fabricius) (Lepidoptera: Noctuidae). The compound exhibited strong growth inhibition in a diet bioassay, with 0.67 p.p.m: and 0.78 p.p.m. of the compound reducing growth by 50% in H. armigera and S. litura neonate larvae, respectively, whereas a growth inhibition of 95% was achieved at 2.36 p.p.m: and 2.41 p.p.m., respectively; this was comparable to azadirachtin treatments used as a control. Aglaroxin A was toxic to various stadia. Nutritional analysis revealed the antifeedant properties of the compound; however, nutritional indices indicated that the reduction in growth of the larvae was not entirely due to starvation, but partly due to the toxic effects of the ingested compound. This was further confirmed in topical treatments. When relative growth rate was plotted against relative consumption rate, the growth efficiency of larvae fed on a diet containing aglaroxin A was significantly less than that of control larvae. These results further indicate that aglaroxin A acts as both antifeedant and chronic toxin. Morphologically deformed or partially pupated insects were obtained after 5th instar larvae were treated with aglaroxin A. Such developmental inhibition during ecdysis was not due to depletion of the moulting hormone, as treated larvae, when provided with exogenous 20‐hydroxyecdysone, did not show any recovery from the effect. However, it is obvious from the present findings that aglaroxin A activity does not absolutely follow the pattern of azadirachtin or the more related compound rocaglamide known in lepidopterans.     

Effect of time and temperature on toxicity of insecticides to insects

The effects of a change in temperature (15–28° C.) on the toxicity of dilute suspensions of DDT to larvae of Aédes aegypti L., as assessed by a photomigration method, depended on the stage of the test during which the temperature was changed. Increase in temperature during exposure to DDT (0.02 p.p.m. for about 1 hr.) increased the toxic action. When larvae were left in the suspensions for the duration of the test (3 hr.-4 days), increase in temperature throughout the test decreased the toxic action of a very low concentration of DDT (0.002 p.p.m.), but had no effect with higher concentrations (0.1–0.2 p.p.m.). Toxic action was greater in larvae held at a low temperature than in larvae held at a high temperature after treatment (0.025 p.p.m. for 3 hr.). Such toxic action was reversible: a change from high to low temperature increased paralysis, and larvae, paralysed at a low temperature, recovered when the temperature was raised.     

Sensitization and habituation of the swimming behavior in ascidian larvae to light

Ascidian larvae of Ciona intestinalis change their photic behavior during the course of development. Newly hatched larvae show no response to a light stimulus at any intensity. At 4 hr after hatching, larvae were induced to start to swimming upon the cessation of illumination, and to stop swimming upon the onset of illumination. At a weaker light intensity (5.0 x 10(-3) J/m (2).s), the larvae showed similar responses to either a single stimulus or repeated stimuli of onset and cessation of light until 10 hr after hatching. At a stronger light intensity (3.2 x 10(-1) J/m(2).s), when the stimulus was repeated, they showed sensitization and habituation of the swimming response. At 3 hr after hatching the larvae failed to show any response to an initial stimulus at any intensity of light, but after several repeated stimuli (sensitization) they showed a swimming response at light intensities above 4.0 x 10(-2) J/m (2).s. At 5 hr and with intensity above 1.0 x 10 (-2) J/m(2).s, the larvae showed photoresponses to the first stimulus, but after several repetitions the larvae failed to stop swimming upon the onset of light (habituation). A repeated series of stimuli at stronger intensities of light caused greater habituation; this habituation was retained for about 1 min. Since the larval central nervous system in Ciona is comprised of only about 100 neurons, learning behavior in ascidian larvae should provide insights for a minimal mechanism of memory in vertebrates.     

Daily variations of antioxidant enzyme and luciferase activities in the luminescent click-beetle Pyrearinus termitilluminans: cooperation against oxygen toxicity

Several lines of investigation have suggested an interplay between bioluminescence (BL) and oxyradical metabolism, mainly in bacteria and beetles. Although not yet confirmed, luminescent beetles seem to be challenged daily by oxidative conditions imposed by higher oxygen absorption necessary to enhance light emission for courtship (adult lampyrids and elaterids) and prey attraction (e.g. Pyrearinus termitilluminans larvae). This work reports the activities of luciferase, superoxide dismutase (SOD), catalase and dehydroascorbate reductase (DHAR) and total glutathione content at different times of the day in the bright prothorax and dim abdomen of larval Pyrearinus termitilluminans (Coleoptera: Elateridae), investigating a possible adjuvant role for luciferase in oxygen detoxification. Luciferase activity in the prothorax was shown to peak at 7 p.m., which is the time when P. termitilluminans larvae light up for prey attraction. In their habitat, P. termitilluminans larvae emit light until 8.30 p.m. However, at 8 p.m., prothorax luciferase activity achieved basal levels and total glutathione content declined to the daily lowest value, possibly resulting from hyperoxidative conditions during this time. Significant increases in the activities of total SOD (28%) and catalase (37%) were observed in the prothorax at 9 p.m., which should minimize the extent of damage from this potentially hazardous period. Prothorax total SOD (42% higher than daily average) and abdomen CuZnSOD (41%) and catalase (95%) activities showed extra peaks at 7-10 a.m., and abdomen DHAR activity was maximal (37%) earlier (4-7 a.m.). These morning increases in antioxidant enzyme activities may be associated with biological events other than bioluminescence, e.g. intense physical activity for digging tunnels and/or digestion of captured preys. These data suggest that oxyradical pathway and bioluminescence are coordinated, especially in the prothorax, to minimize the oxidative stress imposed by higher irrigation of the photocytes with O(2) when P. termitilluminans larvae emit light.     

Toxicity of cypermethrin: hsp70 as a biomarker of response in transgenic Drosophila

Heat shock protein induction is often associated with a cellular response to a harmful environment or to adverse life conditions. The main aims of our study were (1) to evaluate the cytotoxic potential of cypermethrin; and (2) to investigate the suitability of stress-induced heat shock protein Hsp70 as a biomarker for environmental pollutants in transgenic Drosophila melanogaster (Hsp70-lacZ)Bg⁹. Different concentrations of cypermethrin (0.002, 0.2, 0.5 and 50.0 p.p.m.) were mixed with food. Third instar larvae of transgenic Drosophila melanogaster were allowed to feed on these mixtures for different time intervals (2, 4, 6, 12, 24 and 48h). Following feeding, hsp70 induction and tissue damage were evaluated. In the highest concentration treatment group (50 p.p.m.), 100% larval mortality was recorded after 12 h exposure. Hsp70 was found to be induced even at the lowest concentration (0.002 p.p.m.) of the insecticide, while tissue damage was observed in the larvae exposed for 48 h. While an insignificant decline in hsp70 expression was observed in the larvae exposed to cypermethrin at a dietary concentration of 0.002 p.p.m. after 48 h compared with those exposed for 24 h, in the next two higher concentrations of the toxicant, a similar but significant decline in hsp70 expression was evident in the exposed larvae after 48 h. The present study reveals the cytotoxic potential of cypermethrin and further proposes that hsp70 induction in transgenic Drosophila melanogaster could be used as a sensitive biomarker in risk assessment.     

Toxicity of cypermethrin: hsp70 as a biomarker of response in transgenic Drosophila

Heat shock protein induction is often associated with a cellular response to a harmful environment or to adverse life conditions. The main aims of our study were (1) to evaluate the cytotoxic potential of cypermethrin; and (2) to investigate the suitability of stress-induced heat shock protein Hsp70 as a biomarker for environmental pollutants in transgenic Drosophila melanogaster (Hsp70-lacZ)Bg⁹. Different concentrations of cypermethrin (0.002, 0.2, 0.5 and 50.0 p.p.m.) were mixed with food. Third instar larvae of transgenic Drosophila melanogaster were allowed to feed on these mixtures for different time intervals (2, 4, 6, 12, 24 and 48h). Following feeding, hsp70 induction and tissue damage were evaluated. In the highest concentration treatment group (50 p.p.m.), 100% larval mortality was recorded after 12 h exposure. Hsp70 was found to be induced even at the lowest concentration (0.002 p.p.m.) of the insecticide, while tissue damage was observed in the larvae exposed for 48 h. While an insignificant decline in hsp70 expression was observed in the larvae exposed to cypermethrin at a dietary concentration of 0.002 p.p.m. after 48 h compared with those exposed for 24 h, in the next two higher concentrations of the toxicant, a similar but significant decline in hsp70 expression was evident in the exposed larvae after 48 h. The present study reveals the cytotoxic potential of cypermethrin and further proposes that hsp70 induction in transgenic Drosophila melanogaster could be used as a sensitive biomarker in risk assessment.     

Rubidium marking of Anopheles mosquitoes detectable by field-capable X-ray spectrometry

We present a mosquito marking technique suitable for mark-release-recapture that can be used with a hand-held, portable X-ray fluorescence (XRF) spectrometer, which is practical for field measurements. Third instar Anopheles gambiae Giles sensu stricto (Diptera: Culicidae) and Anopheles stephensi Liston larvae were cultured to pupation in water containing rubidium (Rb) Cl at concentrations up to 1000 p.p.m. Rb. Anopheles gambiae larvae survived to adulthood at concentrations as high as 1000 p.p.m. Rb but suffered pupal mortality and reduced adult longevity at high concentrations. We were able to culture An. stephensi at Rb concentrations as high as 300 p.p.m. The presence of Rb in adults was evaluated using a portable XRF analyser, and we were able to reliably detect Rb above background levels in 10-day-old females and 4-day-old males at concentrations causing minimal pupal or adult mortality. We observed that Rb marking was not permanent, and the concentration declined significantly as adults aged. The low cost of labelling with RbCl and the field portability of the spectrometer provide a useful means for labelling mosquitoes via breeding sites or in the laboratory for mark-release-recapture experiments.     

Effects of oxygen deficiency on survival and glycogen content of Chironomus anthracinus (Diptera,Chironomidae) under laboratory and field conditions

Growth and glycogen content of Chironomus anthracinus in Lake Esrom, Denmark was examined during summer stratification in 1992 and 1993. Simultaneously, effects of oxygen deficiency on glycogen utilization and survival were experimentally studied. The population consisted of almost fullgrown 4th instar larvae in 1992 and 2nd and 3rd instar larvae in 1993. Growth rate and glycogen content changed as hypolimnetic oxygen deficiency increased. During a 1st phase of stratification dry weight and glycogen content increased (2nd and 3rd instars) or was almost constant (4th instar) but decreased significantly during the following 2nd phase. This change from growth to degrowth and utilization of endogenous glycogen reserves correlated with a change in the thickness of the microxic layer (<0.2 mg O₂ 1^–1) above the sediment surface. The layer increased from 2–3 m in phase 1 to 4–5 m in phase 2, and we suggest that this deteriorated the oxygen conditions and resulted in a change in larval energy metabolism from fully aerobic during the 1st phase to partly anaerobic in the 2nd phase. During the 2nd phase larval metabolism was estimated at less than 20% of normoxic rate. Experimental exposure of the larvae to anoxia indicated highly different survival of young larvae (2nd and 3rd instars) and older larvae (large 4th instars). The morality of young larvae was 50% after three days in anoxia at 10 °C, whereas only 25% of the older larvae had died after 3–4 weeks under similar conditions. Extending the treatment, however, resulted in increased death rate of the 4th instar larvae with only 10% surviving after seven weeks. The anaerobic metabolism of 4th instar larvae as estimated from glycogen degradation at 10 °C was 5% of normoxia in the interval from 0–5 days but 1.5% in the interval from 20–25 days. It is concluded that survival of C. anthracinus in anoxia is very limited, but traces of oxygen in the environment allowing for faint aerobic metabolism prolong the survival time of the larvae from a few days (2nd and 3rd instars) or a few weeks (4th instar) to probably 3–4 months.     

Effect of mannose-binding lectin from peanut and pea on the stem borer Chilo partellus

A mannose-binding lectin found in vegetative tissues of peanut, Arachis hypogaea, was compared with mannose-binding lectin from pea, Pisum sativum, for toxic effects on larvae of the stem borer Chilo partellus (Swinhoe). After 10 days, the mortality of larvae fed on artificial diet containing 0.5% (m/m) peanut lectin was 46.2%. The mortality of larvae fed on 1.0% peanut lectin was similar (48.1%) but insects were significantly smaller than those of the 0.5% treatment. Larvae of both lectin treatments stopped feeding within three days. Larval size and mortality was not significantly reduced by 0.1% peanut lectin and 1% heat-treated lectin did not show toxic effects. The mannose-binding lectin from pea was not toxic to C. partellus at concentrations up to 1%. Peanut lectin bound to the apical membranes of columnar epithelial cells in the mid-gut of C. partellus. This suggests that peanut lectin has an antinutritive action and that it may protect vegetative tissues of peanut against insect pests.     

Ovicidal and larvicidal effectiveness of several insect growth inhibitors and regulators on the codling moth Cydia pomonella L. (Lep., Tortricidae)

Several insect growth inhibitors (IGIs) and regulators (IGRs) were tested in the laboratory for their ovicidal and larvicidal properties on the codling moth C. pomonella , by dipping apples in solutions of them. The IGIs which block chitin synthesis – diflubenzuron, hexaflumuron and teflubenzuron – were noticeably more effective against eggs than on newborn larvae with preventive ovicidal 50% lethality concentrations (LC₅₀) values of approximately 0.6, 1.3 and 15 p.p.m., respectively, and larvicidal LC₅₀ values of 104, 1208 and 204 p.p.m. Flufenoxuron, on the other hand, is almost as effective on larvae (LC₅₀ : 9.9 p.p.m.) as on eggs (LC₅₀ : 5.4 p.p.m.). Fenoxycarb, an IGR juvenile hormone analogue, acts as an excellent ovicidal product with an LC₅₀ value of 0.05 p.p.m. Tebufenozide, an IGR ecdyson (moulting hormone) agonist, is exclusively larvicidal with an LC₅₀ at 0.4 p.p.m. Methoxyfenozide, an IGR of the same family and currently being developed, acts as effectively on eggs as on larvae with ovicidal and larvicidal LC₅₀ values of about 0.6 and 0.8 p.p.m., respectively. When ovicidal products are applied as a curative treatment on eggs less than 24 h old, their effectiveness is much lower than that obtained from preventive application.     

Pharmacological Induction of Larval Settlement and Metamorphosis in the Blue Mussel Mytilus edulis L.

The blue mussel Mytilus edulis L. is an important aquaculture and fouling species in northern seas. Although the general role of chemical cues for settlement of larvae of the blue mussel has been proposed, few studies have focused on induction of settlement and metamorphosis by pharmacological agents. In this study, the induction of larval settlement of the blue mussel by pharmacological compounds was investigated through a series of laboratory experiments with an aim of identifying artificial cues for laboratory bioassay systems in fouling and antifouling research. Gamma-aminobutiric acid (GABA), dihydroxyphenyl L-alanine (DOPA), isobutyl methylxanthine (IBMX) and acetylcholine chloride (ACH) at 10^{m 7}-10^{m 2} M as well as KCl at 10-40 mM K⁺ in excess of the level in normal seawater were tested for their inductive effect on larval settlement. In filtered seawater (FSW) <9% of the larvae settled after 48 h. Elevated K⁺ and GABA levels had no effect on larval settlement and metamorphosis. DOPA at 10^{m 5} M and IBMX at 10^{m 6}-10^{m 4} M induced 41-83% larval settlement and ACH at 10^{m 7}-10^{m 5} M induced < 40% larval settlement. While the highest settlement rates were observed after 48 h exposure to the chemicals, most of the larvae settled within 24 h. Compounds at concentrations of 10^{m 3}-10^{m 2} M were either toxic to larvae or retarded the growth of the post-larvae shell. Juveniles resulting from induction by lower concentrations of chemicals had a very high survival rate, completed metamorphosis and grew as well as the juveniles that metamorphosed spontaneously. IBMX at 10^{m 6}-10^{m 4} M and L-DOPA at 10^{m 5} M are effective agents for induction of settlement and metamorphosis for future studies using juvenile M. edulis.     

Effects of insect growth regulators on the hairy rose beetle, Tropinota squalida (Col., Scarabeidae)

Abstract:  Saponin extract from alfalfa roots, azadirachtin from the neem seed oil, synthetic ecdysteroid agonist RH-2485, and the juvenoid hydroprene disturb the development and reproduction of Tropinota squalida . Feeding beetles on diets containing 750 p.p.m. saponins, 7.5 p.p.m. RH-2485, and 1.13 p.p.m. azadirachtin reduces their progeny from 51 second instar larvae per female to 24, 15, and 15 larvae, respectively. When the larvae of untreated adults are fed for 1 week on dung with 75 p.p.m. saponins, 50 p.p.m. RH-2485, and 0.45 p.p.m. azadirachtin, the rate of adult emergence drops from 80% (controls) to 20, 0 and 13%, respectively. No adults emerge when the treatment is continued through the second and third larval instars. Two topical treatments of larvae with 0.2  μ g hydroprene decrease the rate of adult emergence from 90 to 11%, and treatments with 2  μ g prevent adult development in all insects. The observed effects warrant testing of azadirachtin, RH-2485, and hydroprene in the field. Several types of their application for the control of T. squalida are suggested.     

Withdrawal rates of DDT from chickens treated with diphenylhydantoin.

Male and female chickens of a broiler-type strain were fed, from 1 day old to 5 weeks of age, diets containing 0, 2.5, or 15.0 p.p.m. (mg/kg) 1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane (p,p'-DDT). Then the diets with pesticide were withdrawn and the chickens were fed dietary levels of diphenylhydantoin (DPH) at 0, 100, or 250 p.p.m. Adipose-tissue and liver samples were obtained on days 0, 10, 20, and 30 following withdrawal of diets with pesticides to determine DPH effect on DDT, 1,1-dichloro-2,2-bis(p-chlorophenyl)ethylene (DDE), and 1,1-dichloro-2,2-bis(p-chlorophenyl)ethane (DDD) levels. DPH had no effect on the concentration of DDT and DDE in adipose tissue; their levels declined at a rate having a half-life value of 16 days. DDD was not detected in adipose tissue. DDT accounted for 87% of the adipose residues on day 0, but 66% of the residues at day 30. DPH had no effect on the concentrations of DDT and DDE in livers of chickens fed 15.0 p.p.m. DDT, but did significantly reduce the levels of DDD by 28 and 54% for levels of 100 and 250 p.p.m. DPH, respectively. The similarity of these data to studies on dairy cows and humans, and the dissimilarity to data from rat studies were discussed.     

Combined effect of salinity and temperature on Spirorbis spirorbis L. and Circeus spirillum L. larvae from the White Sea

The combined effect of salinity and temperature on Spirorbis spirorbis L. and Circeus spirillum L. larvae from the White Sea was studied in the laboratory experiments. In the White Sea, S. spirorbis is distributed through the depth of 1-20 m and is affected by all varieties of fluctuations in salinity and temperature. C. spirillum lives in more wide range of depths 1-55 m and is more stenohaline. S. spirorbis larvae are sufficiently more resistant to the low salinity (10‰) than C. spirillum larvae. Both species are stenothermic. Highest survivorship of S. spirorbis larvae was marked under 5 °C in all experimental salinities. Under temperature treatments of 10-15 °C, the larval survivorship was sufficiently restricted in all salinities. Highest survivorship of C. spirillum larvae was also marked under 5 °C but in more narrow salinity range.The number of larvae undergoing metamorphosis in both species was very low, only about 10% of the total number. Highest number of successful attachments in both species was marked in high salinities (25-30‰) and does not exceed 25% of survivors. Experimental data suggests that salinity and temperature affect directly general survivorship of the larvae and secondary-attachment and metamorphosis processes.     

The p53 tumor suppressor gene. A preliminary clinical study in breast cancer patients.

p53 was originally considered to be a nuclear oncogene, but several convergent lines of research have indicated that the wild-type gene functions as a tumor suppressor gene negatively regulating the cell cycle. Mutations in the p53 gene have been detected in many tumor types and seem to be the most common genetic alterations in human cancer. In this preliminary study, sera of 92 patients (pts) with breast disease were analyzed for the presence of the mutant p53 protein (mp53) with a selective immunoenzyme assay employing a monoclonal antibody (PAb 240) specific for the majority of mammalian m p53 but not for the wild-type protein. Of the 10 patients with benign breast disease, only two (20%) showed detectable m p53 levels in the serum. In the breast cancer group, sera from 7 of the 30 pts (23%) without lymph node involvement were positive for m p53, as were 7 out of the 45 pts (15%) with metastatic lymph nodes and 1 out of the 7 pts (14%) with disseminated disease. The specificity of m p53 assay evaluated in 20 healthy controls was 100%. These preliminary results showed that serum positivity for m p53 is not related to breast disease extension. Further studies to assess the utility of m p53 as a possible prognosis factor in breast cancer are currently in progress.     

The naturally derived insecticide spinosad is highly toxic to Aedes and Anopheles mosquito larvae

Spinosad is a naturally derived biorational insecticide with an environmentally favourable toxicity profile, so we investigated its potency against mosquito larvae (Diptera: Culicidae). By laboratory bioassays of a suspension concentrate formulation of spinosad (Tracer), the 24 h lethal concentration (LC50) against Aedes aegypti (L.) third and fourth instars was estimated at 0.025 p.p.m. following logit regression. The concentration-mortality response of third- and fourth-instar Anopheles albimanus Weidemann did not conform to a logit model. The LC50 value of spinosad in Anopheles albimanus was 0.024 p.p.m. by quadratic linear regression. A field trial in southern Mexico demonstrated that spinosad 1 p.p.m. compared with the standard temephos (Abate) 1% granules 100 g/m3 water prevented Ae. aegypti breeding in plastic containers of water for 8 weeks; at 10 p.p.m. spinosad prevented breeding for > 22 weeks. In another field trial, spinosad at 5 p.p.m. and temephos both completely eliminated reproduction of Ae. aegypti for 13 weeks. In contrast, the bacterial insecticide Bacillus thuringiensis var. israelensis (Bti, Vectobac) AS) performed poorly with just 2 weeks of complete inhibition of Ae. aegypti breeding. Spinosad also effectively prevented breeding of Culex mosquitoes and chironomids in both trials to a degree similar to that of temephos. We conclude that spinosad merits evaluation as a replacement for organophosphate or Bti treatment of domestic water tanks in Mesoamerica. We also predict that spinosad is likely to be an effective larvicide for treatment of mosquito breeding sites.     

Experimental Trichinella infection in seals

The susceptibility of seals to infection with Trichinella nativa and the cold tolerant characteristics of muscle larvae in seal meat were evaluated. Two grey seals, Halichoerus grypus, were inoculated with 5000 (100 larvae/kg) T. nativa larvae and two grey seals with 50000 (1000 larvae/kg). One seal from each dose group and two control seals were killed at 5 and 10 weeks post-inoculation (p.i.). At 5 weeks p.i., infection was established in both low and high dose seals with mean larval densities of 68 and 472 larvae per gram (lpg), respectively, using eight different muscles for analyses. At 10 weeks p.i., mean larval densities were 531 and 2649 lpg, respectively, suggesting an extended persistence of intestinal worms. In seals with high larval density infections, the distribution of larvae in various muscles was uniform, but in one seal with a low larval density infection, predilection sites of larvae included muscle groups with a relative high blood flow, i.e. diaphragm, intercostal and rear flipper muscles. Trichinella-specific antibody levels, as measured by ELISA, increased during the 10 week experimental period. Infected seal muscle was stored at 5, -5 and -18 degrees C for 1, 4 and 8 weeks. Muscle larvae released from stored seal muscle by artificial digestion were inoculated into mice to assess viability and infectivity. Larvae from seal muscle 10 weeks p.i. tolerated -18 degrees C for 8 weeks but larvae from seal muscle 5 weeks p.i. tolerated only 1 week at -18 degrees C, supporting the hypothesis that freeze tolerance increases with the age of the host-parasite tissue complex. The expressed susceptibility to infection, extended production of larvae, antibody response and freeze tolerance of T. nativa in seals are new findings from the first experimental Trichinella infection in any marine mammal and suggest that pinnipeds (phocids, otariiids or walrus) may acquire Trichinella infection by scavenging even small amounts of infected tissue left by hunters or predators.