Prospects for using pyroproxyfen-treated targets for tsetse control

Using¹⁴C cholesterol as a marker a positive correlation was established between the amount of oil (a chlorinated n-alkane containing 43–46% chlorine, ‘cereclor S45’) picked up by an adult tsetse fly exposed by tarsal contact to a treated surface and the duration of such exposure. Only a poor uptake was achieved from netting surfaces treated with less than 50% oil in acetone. Terylene netting treated with radioactive pyriproxyfen, [1-methyl-2-(4-phenoxyphenoxy)ethoxy] pyridine, dissolved in cereclor, was exposed in the field for a year. After 9 months 20% of the original radioactivity remained and was shown to be 95% authentic pyriproxyfen. Brief tarsal contact (up to 5 seconds) with such netting, by adult females ofGlossina morsitans morsitans Westwood, reduced the viability of their offspring to 28–43% of untreated control values. The effect was greatest in the reproductive cycle immediately following contact. Between 10 and 12 months after treatment of the fabric the radioactivity fell to only 7% of the original level but was associated mainly (>80%) with intact pyriproxyfen. Exposure of female flies to this netting resulted in a positive correlation between the duration of exposure and the extent of suppression of offspring viability, such that 2 min was sufficient to reduce offspring viability to zero for the life of the female. Traps or targets impregnated with conventional formulations of pyrethroids to kill tsetse would have lost all their activity by this time. Results are discussed in terms of the prospects for using pyriproxyfen-treated targets to sterilize female tsetse directly and also indirectly through the contamination of males prior to mating through contact with such targets.     

Effects of the juvenile hormone mimic pyriproxyfen on female reproduction and longevity in the butterfly Bicyclus anynana

Female Bicyclus anynana butterflies given pyriproxyfen, a mimic of juvenile hormone, exhibited increased egg‐laying rates and early fecundity, but reduced longevity compared with control animals. Thus, pyriproxyfen application yielded antagonistic effects on different components of fitness, possibly demonstrating a juvenile hormone‐mediated trade‐off between present and future reproduction. Lifetime fecundity and egg size, however, showed no consistent response to pyriproxyfen, with lifetime fecundity being increased or decreased and egg size being reduced in one out of four experiments only. Females were most sensitive to pyriproxyfen around the onset of oviposition, coinciding with naturally increasing juvenile hormone titers in other Lepidoptera. Amounts between 1 and 10 µg pyriproxyfen were found to be effective, with, however, pronounced differences among experiments. This is attributed to differences in assay conditions. High pyriproxyfen concentrations (100 µg) as well as repeated applications of smaller amounts did not affect reproductive traits, but tended to reduce longevity.     

Does reproductive investment change with age in tsetse flies, Glossina morsitans morsitans (Diptera: Glossinidae)?

1. Viviparous insects such as tsetse ( Glossina spp.) provide unusual opportunities to compare age-related changes in the proportion of maternal resources transferred to offspring.
2. In laboratory populations of Glossina morsitans morsitans the survival of females was high for the first 60 days of adult life but declined rapidly thereafter.
3. Average longevity did not differ significantly between mated and unmated females (93·6 and 90 days, respectively).
4. Nutritional state in terms of fat content and residual dry mass did not decline with adult female age.
5. The fecundity of mated females was constant for the first 60 days of adult life and declined only slightly thereafter.
6. Offspring size did not change towards the end of the adult female lifespan and there was no evidence of an increase in the allocation of resources to reproduction in older females.
7. Results contrast with those obtained recently for vertebrates and may indicate that age-related changes in offspring size in Glossina are not adaptive, or that so few females reach old age under natural conditions that there is no selection for a strategy of terminal investment.     

Precocene-induced sterility in F1 generation of Glossina morsitans morsitans

Precocene treatment does not disrupt the events of reproduction in Glossina morsitans morsitans or induce any apparent changes in treated tsetse. However, some females of the F₁ generation are either sterile or show retardations in follicle development. Sterility is not reversed spontaneously or with juvenile hormone analogues. The critical period for precocene action is related to each ovulation. The corpora allata of precocene-treated tsetse are normal, but those of F₁ sterile females are degenerate. The occurrence of retardation has enabled the characterisation of stages in follicle development in G. m. morsitans.     

The effect of colour on the numbers, age and nutritional status of Glossina tachinoides (Diptera: Glossinidae) attracted to targets

Abstract The effect of colour on the attractiveness of cloth targets for G.tachinoides Westwood was investigated, using electrified nets. The ages and nutritional states of tsetse caught by different targets were also studied. With targets of cloth and mosquito netting panels, phthalogen blue was the most attractive colour, and yellow the least, with black, red, violet and white intermediate. The strongest landing responses in females occurred on UV-reflecting white cloth. In males, landing was high with all colours except yellow. Colour combination targets were no better than all-blue targets when mosquito netting side-panels were present; in their absence, blue-and-white targets were over twice as good as all-blue targets. No consistent differences in ages or nutritional states were found between tsetse caught by differently coloured targets, but those landing directly on the cloth portion of a target had lower fat reserves than those intercepted on an adjacent netting panel.     

Effects of pyriproxyfen on ovarian development and fecundity ofRhagoletis pomonella flies

Three-day old female apple maggot flies,Rhagoletis pomonella (Walsh), were topically exposed to different doses (0.1, 1, 10, and 100 μg per fly) of a juvenile hormone mimic, pyriproxyfen, in the laboratory. Pyriproxyfen had little lethal effect on females except at the extremely high dose of 100 μg. It also had no significant effect on egg viability of treated females. A non-lethal dose of 1 μg per fly did, however, enhance significantly the fecundity (egg production) as well as the ovarian development (number of eggs in ovaries and length of egg folicles) of treated flies. We conclude that pyriproxyfen could be a useful aid in exploring endocrine regulation of feeding and reproductive physiology behavior inR. pomonella, about which current knowledge is scant.     

Responses of tsetse flies, Glossina morsitans morsitans and Glossina pallidipes, to baits of various size

Recent studies of Palpalis group tsetse [Glossina fuscipes fuscipes (Diptera: Glossinidae) in Kenya] suggest that small (0.25 × 0.25 m) insecticide-treated targets will be more cost-effective than the larger (≥1.0 × 1.0 m) designs currently used to control tsetse. Studies were undertaken in Zimbabwe to assess whether small targets are also more cost-effective for the Morsitans group tsetse, Glossina morsitans morsitans and Glossina pallidipes. Numbers of tsetse contacting targets of 0.25 × 0.25 m or 1.0 × 1.0 m, respectively, were estimated using arrangements of electrocuting grids which killed or stunned tsetse as they contacted the target. Catches of G. pallidipes and G. m. morsitans at small (0.25 × 0.25 m) targets were, respectively, ～1% and ～6% of catches at large (1.0 × 1.0 m) targets. Hence, the tsetse killed per unit area of target was greater for the larger than the smaller target, suggesting that small targets are not cost-effective for use against Morsitans group species. The results suggest that there is a fundamental difference in the host-orientated behaviour of Morsitans and Palpalis group tsetse and that the former are more responsive to host odours, whereas the latter seem highly responsive to visual stimuli.     

Biotypes B and Q of Bemisia tabaci and their relevance to neonicotinoid and pyriproxyfen resistance

Resistance monitoring for Bemisia tabaci field populations to the juvenile hormone mimic, pyriproxyfen, was conducted from 1996 to 2003 in commercial cotton fields in two areas of Israel: the Ayalon Valley (central Israel) and the Carmel Coast (northwestern Israel). Although the use of pyriproxyfen ceased in these areas in 1996-1997 (because of the resistance), resistance levels to pyriproxyfen declined to some extent in the fields but remained quite stable, and the susceptibility has not been totally restored. Two strains of B. tabaci collected from the Ayalon Valley in the late 1999 and 2002 cotton seasons (AV99L, AV02L) were assayed for their susceptibility to pyriproxyfen at F1, and subsequently a line of each strain was kept under controlled conditions without exposure to insecticides. After maintenance of more than 20 generations under laboratory conditions, the resistance to pyriproxyfen in the untreated strains substantially declined. This decline was concurrent with a replacement of Q biotype by B-type under non-insecticidal regimes; apparently B biotype was more competitive than the pyriproxyfen-resistant Q-type. Selection under controlled conditions with neonicotinoids on these B. tabaci strains resulted in continued pyriproxyfen resistance, predominantly of Q biotype. Based on our data, applications of either pyriproxyfen or neonicotinoids may select for biotype Q, which would survive to a greater degree where these insecticides are applied.     

Feeding frequency in relation to reproduction in Glossina morsitans morsitans and G.pallidipes

Abstract Measurements of residual haematin in males of Glossina morsitans morsitans Westwood reared in the laboratory at 25^oC suggest that blood meal digestion is completed in 4 or 5 days after feeding. However, a high proportion of haematin is present as faecal matter 2 days after feeding and it is concluded that digestion is completed sooner than indicated by the regression of logio haematin on time. Therefore, low levels of residual haematin in field-caught tsetse provide no indication of the frequency with which they feed. For this reason the effects of feeding frequency upon various reproductive parameters in the laboratory have been examined. It is concluded that the best performance is achieved by G.m. morsitans females which ingest four blood meals per inter-larval period and that for a similar performance in G.pallidipes five blood meals are required. The extent to which such feeding frequencies are a reflection of feeding activity in the field are discussed in terms of the biochemical requirements to maintain a reproductive adult female tsetse in positive energy balance.     

A behavioural bioassay to identify attractive odours for Glossinidae

1. A behavioural bioassay, based on antennal movement responses, was developed using Glossina morsitans morsitans Westwood for screening chemical attractancy to tsetse. 2. Chemicals found to be attractive to male tsetse were acetone, formaldehyde, methylethylketone, methylvinylketone, 1-octen-3-ol and pentanal but not acetophenone, hexanal, lactic acid or urea. 3. Female tsetse also responded to all these chemicals in a similar fashion. Overall responses of females were, however, less than those of males. 4. These laboratory bioassay findings agree with field observations on tsetse responses to certain chemical odours. Therefore this behavioural bioassay should be a useful laboratory test procedure for screening attractants.     

Specificity of contact sex pheromones in tsetse flies, Glossina spp.

ABSTRACT. Interspecific mating tests among seven tsetse species covering the 'morsitans' and 'palpalis' groups showed a degree of reaction that extends beyond the traditional taxonomic groupings. Tests with either live or dead females as targets produced essentially similar results which shows that differing responses of males are largely the consequence of differences in composition of the female surface cuticular paraffins and not of female behaviour. Gas chromatographic (GLC) analysis of cuticular paraffins tends to confirm this view. Responses of males of different species to dead conspecific males baited with the most active synthetic G.morsitans pheromone were variable. Comparison of behavioural responses and of GLC profiles of cuticular paraffins suggest that the species tested do not share a pheromone structure in common with G.morsitans. Mating tests with G.morsitans males using dead conspecific males baited with a variety of synthetic compounds provided evidence of the molecular configuration required to elicit a mating response. Synthetic 15,19-dimethyltritriacontane induced a suboptimal response in male G.morsitans but is a likely candidate for a sex pheromone in G.austeni. Although not conclusive, a comparison of behavioural evidence and GLC data has permitted speculation on the nature of sex pheromones in tsetse of the 'palpalis' group.     

Pyriproxyfen enhances the immunoglobulin G immune response in mice

Pyriproxyfen is a juvenile hormone mimic of vital importance for insect development with little risk to humans. This study was performed to investigate whether large doses of pyriproxyfen affect the immune response in mammals. Mice were immunized thrice with ovalbumin in 5% ethanol, with or without pyriproxyfen or alum. Large doses of pyriproxyfen (9 or 15 mM) significantly enhanced specific total IgG immune response. This enhancement was no longer present 24 hr after treatment with pyriproxyfen. These results suggest that pyriproxyfen is a safe chemical. Moreover, pyriproxyfen induced higher titers of IgG2a and enhanced tumor necrosis factor‐alpha and gamma‐interferon responses whereas alum induced IgG1 with enhanced interleukin‐4 and ‐10. These observations indicate that the mechanism of immune enhancement by pyriproxyfen may differ from that of alum.     

Biorational insecticides: mechanism and cross-resistance

Potency and cross-resistance of various biorational insecticides, exemplified by the whitefly Bemisia tabaci, have been studied. Bemisia tabaci were exposed to the juvenile hormone mimic pyriproxyfen for the past 12 years resulting in an over 2,000-fold resistance, but there was no appreciable cross-resistance with the benzoylphenyl urea novaluron. Similarly, no cross-resistance was found between pyriproxyfen and the two neonicotinoids, acetamiprid and imidacloprid. On the other hand, a slight cross-resistance of 5-13-fold was observed with another neonicotinoid thiamethoxam. Among the neonicotinoids, a resistant strain of B. tabaci to thiamethoxam (approximately 100-fold) showed no appreciable cross-resistance to either acetamiprid or imidacloprid, while another strain 500-fold resistant to thiamethoxam resulted in a mild of 4-6-fold resistance to acetamiprid and imidacloprid. In other assays, B. tabaci strain resistant to thiamethoxam (approximately 100-fold) had no cross-resistance to pyriproxyfen. Our findings indicate that no appreciable cross-resistance was observed between the benzoylphenyl urea novaluron, the juvenile hormone mimic pyriproxyfen, and the neonicotinoids acetamiprid and imidacloprid. Hence, these compounds could be used as components in insecticide resistance management programs.     

Juvenile hormone mimics as effective sterilants for the tsetse fly Glossina morsitans morsitans

The development of puparia of Glossina morsitans morsitans Westwood was disrupted by topical applications of the juvenile hormone mimics S-methoprene (the resolved enantiomer of 11-methoxy-3,7,11-trimethyl-2,4-dodecadienoic acid 1-methyl ester) (Zoecon), S21149 (propionaldoxime-0-4-phenoxyphenoxyethylether) (Sumitomo), or S31183 (2-[1-methyl-2-(4-phenoxyphenoxy)ethoxy]pyridine) (Sumitomo) dissolved in acetone. Puparia so treated during the first 4 days of life suffered developmental abnormalities, the severity of which were dose-dependent. Similarly, puparia produced by adult females treated with these compounds were abnormal. Dose-response data showed that effects were greatest with S31183 and least with S-methoprene. Abnormalities in the form of abdominal lesions and wing crumpling were typical of flies emerging from puparia produced by S-methoprene-treated females. However, arrested development at the red eye and pigmented seta stage within the puparium were typical of offspring of females treated with S21149 and S31183. A dose of 2 micrograms per female of S31183 was sufficient to prevent emergence of offspring produced for the rest of the life of the fly. The same dose resulted in partial recovery of females treated with S21149 some 18 days following treatment. Treatment with 2 micrograms S-methoprene did not suppress completely the production of normal offspring and recovery was complete some 27-35 days after treatment. Exposure of males to 20 micrograms S31183 did not impair their ability to inseminate females; transfer of material during copulation was sufficient to prevent the production of viable offspring by their mates.(ABSTRACT TRUNCATED AT 250 WORDS)     

Standardizing visual control devices for tsetse flies: West African species Glossina tachinoides, G. palpalis gambiensis and G. morsitans submorsitans

Here we describe field trials designed to standardize tools for the control of Glossina tachinoides, G. palpalis gambiensis and G.morsitans submorsitans in West Africa based on existing trap/target/bait technology. Blue and black biconical and monoconical traps and 1 m(2) targets were made in either phthalogen blue cotton, phthalogen blue cotton/polyester or turquoise blue polyester/viscose (all with a peak reflectance between 450-480 nm) and a black polyester. Because targets were covered in adhesive film, they proved to be significantly better trapping devices than either of the two trap types for all three species (up to 14 times more for G. tachinoides, 10 times more for G. palpalis gambiensis, and 6.5 times for G. morsitans submorsitans). The relative performance of the devices in the three blue cloths tested was the same when unbaited or baited with a mixture of phenols, 1-octen-3-ol and acetone. Since insecticide-impregnated devices act via contact with flies, we enumerated which device (traps or targets) served as the best object for flies to land on by also covering the cloth parts of traps with adhesive film. Despite the fact that the biconical trap proved to be the best landing device for the three species, the difference over the target (20-30%) was not significant. This experiment also allowed an estimation of trap efficiency, i.e. the proportion of flies landing on a trap that are caught in its cage. A low overall efficiency of the biconical or monoconical traps of between 11-24% was recorded for all three species. These results show that targets can be used as practical devices for population suppression of the three species studied. Biconical traps can be used for population monitoring, but a correction factor of 5-10 fold needs to be applied to captures to compensate for the poor trapping efficiency of this device for the three species.     

The Insect Growth Regulator Pyriproxyfen Terminates Egg Diapause in the Asian Tiger Mosquito,Aedes albopictus

The Asian tiger mosquito, Aedes albopictus, is a highly invasive mosquito species that transmits chikungunya and dengue. This species overwinters as diapausing eggs in temperate climates. Early diapause termination may be a beneficial strategy for winter mosquito control; however, a mechanism to terminate the diapause process using chemicals is not known. We tested the hypothesis that a hormonal imbalance caused by the administration of juvenile hormone analog would terminate egg diapause in A. albopictus. We tested the insect growth regulator pyriproxyfen on all developmental stages to identify a susceptible stage for diapause termination. We found that pyriproxyfen treatment of mosquito eggs terminated embryonic diapause. The highest rates of diapause termination were recorded in newly deposited (78.9%) and fully embryonated (74.7%) eggs at 0.1 and 1 ppm, respectively. Hatching was completed earlier in newly deposited eggs (25–30 days) compared to fully embryonated eggs (71–80 days). The combined mortality from premature diapause termination and ovicidal activity was 98.2% in newly deposited and >98.9% in fully embryonated eggs at 1 ppm. The control diapause eggs did not hatch under diapausing conditions. Pyriproxyfen exposure to larvae, pupae and adults did not prevent the females from ovipositing diapausing eggs. There was no effect of pyriproxyfen on diapausing egg embryonic developmental time. We also observed mortality in diapausing eggs laid by females exposed to pyriproxyfen immediately after blood feeding. There was no mortality in eggs laid by females that survived larval and pupal exposures. In conclusion, diapausing eggs were the more susceptible to pyriproxyfen diapause termination compared to other life stages. This is the first report of diapause termination in A. albopictus with a juvenile hormone analog. We believe our findings will be useful in developing a new control strategy against overwintering mosquito populations.     

Infections with immunogenic trypanosomes reduce tsetse reproductive fitness: potential impact of different parasite strains on vector population structure

The parasite Trypanosoma brucei rhodesiense and its insect vector Glossina morsitans morsitans were used to evaluate the effect of parasite clearance (resistance) as well as the cost of midgut infections on tsetse host fitness. Tsetse flies are viviparous and have a low reproductive capacity, giving birth to only 6-8 progeny during their lifetime. Thus, small perturbations to their reproductive fitness can have a major impact on population densities. We measured the fecundity (number of larval progeny deposited) and mortality in parasite-resistant tsetse females and untreated controls and found no differences. There was, however, a typanosome-specific impact on midgut infections. Infections with an immunogenic parasite line that resulted in prolonged activation of the tsetse immune system delayed intrauterine larval development resulting in the production of fewer progeny over the fly's lifetime. In contrast, parasitism with a second line that failed to activate the immune system did not impose a fecundity cost. Coinfections favored the establishment of the immunogenic parasites in the midgut. We show that a decrease in the synthesis of Glossina Milk gland protein (GmmMgp), a major female accessory gland protein associated with larvagenesis, likely contributed to the reproductive lag observed in infected flies. Mathematical analysis of our empirical results indicated that infection with the immunogenic trypanosomes reduced tsetse fecundity by 30% relative to infections with the non-immunogenic strain. We estimate that a moderate infection prevalence of about 26% with immunogenic parasites has the potential to reduce tsetse populations. Potential repercussions for vector population growth, parasite-host coevolution, and disease prevalence are discussed.     

Improving the cost-effectiveness of visual devices for the control of riverine tsetse flies, the major vectors of human African trypanosomiasis

Control of the Riverine (Palpalis) group of tsetse flies is normally achieved with stationary artificial devices such as traps or insecticide-treated targets. The efficiency of biconical traps (the standard control device), 1×1 m black targets and small 25×25 cm targets with flanking nets was compared using electrocuting sampling methods. The work was done on Glossina tachinoides and G. palpalis gambiensis (Burkina Faso), G. fuscipes quanzensis (Democratic Republic of Congo), G. f. martinii (Tanzania) and G. f. fuscipes (Kenya). The killing effectiveness (measured as the catch per m(2) of cloth) for small targets plus flanking nets is 5.5-15X greater than for 1 m(2) targets and 8.6-37.5X greater than for biconical traps. This has important implications for the costs of control of the Riverine group of tsetse vectors of sleeping sickness.     

Parturition hormone in the tsetse Glossina morsitans: activity in reproductive tissues from other species and response of tsetse to identified neuropeptides and other neuroactive compounds

Parturition hormone (PH) activity is present not only in the uterus of the tsetse Glossina morsitans but also in the oviducts of Bombyx mori and Schistocerca gregaria, as well as the ejaculatory duct of S. gregaria males. Activity thus appears to be present in the reproductive ducts of diverse insect taxa. To determine whether any of the common insect neuropeptides are capable of mimicking the effect of PH, 35 identified neuropeptides and analogs were evaluated for PH activity. Modest PH activity was observed for only high doses of proctolin and a pyrokinin analog, thus suggesting that PH is unlikely to be closely related to any of the identified neuropeptides tested. While proctolin was highly effective in stimulating contractions of the S. gregaria oviduct, the extract from the tsetse uterus elicited only a weak response in this bioassay. PH activity was, however, effectively mimicked with an injection of 8 bromo-cyclic GMP, thus suggesting a potential role for this cyclic nucleotide in mediating the PH response. Pregnant females were responsive to PH, other neuropeptides and cyclic nucleotides only when females were neck-ligated. In intact females, the brain can presumably override the stimulation provided by the active compounds.