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Mode of action of lufenuron on larval cat fleas (Siphonaptera: Pulicidae).

Adult cat fleas, Ctenocephalides felis (Bouché), were fed suboptimal in vitro concentrations of lufenuron in blood to allow hatching of flea larvae for cytological study. At concentrations of 0.125, 0.25, and 0.5 ppm, larval hatch was 64, 15, and 4%, respectively. Larvae hatching from eggs laid by adults fed lufenuron at concentrations of 0.025, 0.08, or 0.125 ppm did not differ significantly from the control. However, many larvae from the 0.08-ppm group and higher concentrations died during the 1st instar. Examination of these larvae revealed that they were dying from desiccation caused by bleeding from microscopic lesions in the cuticle or the inability to complete the molt to the next instar. Electron micrographs showed that lufenuron often disrupted formation of the endocuticle resulting in the deposition of an amorphous mass of randomly oriented chitin microfibrils. Other larvae formed normal endocuticle but were unable to digest the old endocuticle or produce new procuticle after apolysis. Failure of larvae to digest old cuticle or form new cuticle was caused by degeneration of the epidermal cells needed for the synthesis of molting fluid and chitin.

Animals↗

Chorion formation and ultrastructure of the egg of the cat flea (Siphonaptera: Pulicidae).

Oocyte development in adult female cat fleas, Ctenocephalides felis (Bouché), was studied by light and electron microscopy to determine the formation and ultrastructural morphology of the eggshell. As oocytes develop, somatic follicle cells from the lining of the ovariole migrate around the oocytes. The follicle cells produce electron-dense granules that form the vitelline membrane around the developing oocyte. Subsequently, electron-lucent granules containing an electron-dense core (precursors of the chorion) are produced from the rough endoplasmic reticulum that appear as dilated and clear linear clefts in the cytoplasm of the follicle cells. Exocytosis and coalescence of the granules around the oocyte as the follicle cells disintegrate give rise to the chorion. The chorion was found to consist of 4 distinct layers. The external surface of the egg shell consists of a particulate layer approximately 760 nm thick, composed of an electron-lucent layer of widely dispersed granules. Embedded in this layer are electron-dense spheres that project above the surface of this granular layer. Beneath this outermost layer is a band of electron-dense material, consisting of densely packed granules and is half as thick as the outer particulate layer. The 3rd layer consists of relatively thick, weakly laminated chorion, with a felt-like appearance due to a meshwork of microfibrils. Projections of this network of microfibrils form pillars that attach this layer to a thin relatively compact 4th or basal layer. The pillars and the air-filled cavities lying between the 3rd and 4th chorionic layers constitute the chorionic meshwork known as the palisades or trabecular layer that form the major respiratory organ of the eggshell. The trabecular layer is connected to the external environment by means of the lateral and anterior aeroplyes. The vitelline membrane lies between the chorion and oocyte and is a granular, uniform, moderately electron-dense layer measuring approximately 260 nm thick. The micropyle at the posterior of the flea egg consists of a rosette of 50-80 apertures and possesses an internal electron-dense plug between the chorion and the vitelline membrane. An aeropyle at the anterior end of the egg consists of a rosette of 40-50 apertures. An inconspicuous aeropyle appears as a cluster of hexagonal or polygonal-shaped plaques on the lateral surface of the chorion. Each plaque contains 3-8 pores.

Animals↗

Emergence, survival, and fecundity of adult cat fleas (Siphonaptera: Pulicidae) exposed as pupae to juvenile hormone mimics.

Cat flea, Ctenocephalides felis felis (Bouché), adults exposed to sprays of methoprene, pyriproxyfen, or fenoxycarb as cocooned pupae emerged approximately 1 d earlier than adults from water-treated control pupae. Mortality of adult fleas, after exposure to juvenile hormone mimics as pupae, was increased over that of controls. Females had higher mortality than males within the first 48 h of feeding. Fecundity of females exposed as pupae to juvenile hormone mimics was not different from that of controls. Early emergence of preemerged adults from treated cocoons is discussed along with reasons for higher female susceptibility to juvenile hormone mimics.

Animals↗

Identification of two cDNAs encoding synaptic vesicle protein 2 (SV2)-like proteins from epithelial tissues in the cat flea, Ctenocephalides felis.

Two distinct cDNAs that appear to encode proteins in the synaptic vesicle-2 (SV2) family were identified as expressed sequence tags from a Ctenocephalides felis hindgut and Malpighian tubule (HMT) cDNA library. To date, SV2 proteins have been described only in vertebrates, and have been detected only in synaptic vesicles in neuronal and endocrine tissues, where they are thought to regulate synaptic vesicle exocytosis. The cDNAs for the C. felis SV2-like proteins SVLP-1 and SVLP-2 encode predicted full-length proteins of 530 and 726 amino acids, respectively. Of characterized proteins, the SVLP protein sequences were most similar to rat SV2B. Northern blot analysis revealed that both mRNAs were up-regulated in larval stages that feed and in adults after feeding, and were expressed primarily or exclusively in the HMT tissues in adult fleas. These results suggest that the flea SVLP-1 and SVLP-2 gene products may have roles that are specific for the HMT tissues, and may differ in function from vertebrate SV2 proteins.

Amino Acid Sequence↗

Amitraz: a tick and flea repellent and tick detachment drug.

A topical formulation of amitraz (Mitaban Liquid Concentrate, The Upjohn Company, Kalamazoo, Michigan, U.S.A.) was evaluated as a tick repellent and detachment agent, and flea repellent. The diluted liquid concentrate (250 p.p.m. active drug) was topically applied as a single treatment to dogs; the concentration was identical to the rate recommended for treatment of demodicosis and scabies. Brown dog tick (Rhipicephalus sanguineus) and American dog tick (Dermacentor variabilis) populations were eliminated and repelled. Repellent activity (92-95%) was observed against R. sanguineus for 2 weeks post-treatment; the treatment was moderately active (63%) during the third week, and at 4 weeks post-treatment the drug was inactive. Established R. sanguineus populations were also treated, and the diluted liquid concentrate had 100% tick-detachment efficacy. Repellent activity (99%) was also detected against D. variabilis; the activity was monitored for only 7 days. The ectoparasiticide had low to moderate flea (Ctenocephalides felis) repellent activity (42%) for 4 days post-treatment; thereafter the treatment was ineffective. Side-effects were not observed in any of the dogs treated with amitraz, or the placebo.

Administration, Topical↗

Ctenophthalmus harputus, a new Spalax flea from Turkey.

Ctenophthalmus harputus sp. n. is described from the mole rat Spalax ehrenbergi Nehring collected near Elaziğ city in eastern Turkey. This new flea species is classified in the Ctenophthalmus (Palaeoctenophthalmus) fissurus Wagner group. A key is given to the six species of Spalax fleas (Siphonaptera: Hystrichopsyllidae, Ctenophthalmus spp.) recorded from Turkey.

Animals↗

Rickettsia felis: a new species of pathogenic rickettsia isolated from cat fleas.

A flea-borne rickettsia, previously referred to as ELB, has been implicated as a cause of human illness. Using sequence data obtained from a fragment of the citrate synthase gene, we compared ELB, Rickettsia australis, R. rickettsii, and R. akari with the louse-borne R. prowazekii. We tallied 24 base pair differences between ELB and R. prowazekii and 25 between R. rickettsii and R. prowazekii; there were 30 base pair differences between R. australis and R. prowazekii and 29 between R. akari and R. prowazekii. We observed 32 differences between Rickettsia typhi and ELB. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblot analyses of ELB, with typing sera against R. typhi indicate that ELB surface antigens are more closely related to the flea-borne R. typhi than to the mite-borne R. akari. On the basis of the results of citrate synthase gene sequence comparisons, as well as previous comparisons with 16S rRNA and 17-kDa-protein gene segments, we found that ELB is sufficiently genetically distinct from other rickettsiae to be designated a new species, Rickettsia felis.

Animals↗

Prevalence of Rickettsia felis in the fleas Ctenocephalides felis felis and Ctenocephalides canis from two Indian villages in Sao Paulo Municipality, Brazil.

We evaluated the presence of Rickettsia infection among fleas collected on domestic dogs in two Guarani Indian communities in the suburban area of São Paulo Municipality, Brazil. A total of 114 Ctenocephalides felis felis and 47 Ctenocephalides canis were collected from 40 dogs. A total of 41 C. felis felis (36.0%) and 9 C. canis (19.1%) fleas yielded expected bands by PCR, which were all shown by DNA sequencing to be indentical to the corresponding sequence of a fragment of the Rickettsia felis gltA gene deposited in GenBank. The overall prevalence of R. felis was 31.0% (49/161).

Animals↗

Molecular cloning and overexpression of fleA gene encoding a fucose-specific lectin of Aspergillus oryzae.

A protein from the cell lysate of Aspergillus oryzae was purified by column chromatography immobilized with a ferrichrysin (Fcy), which is one of the siderophores of A. oryzae. It is produced only in an iron-deficient culture and its molecular weight is estimated as 35,000 by SDS-PAGE. Two internal amino acid sequences of the protein obtained by lysylendopeptidase digestion were analyzed. Molecular cloning shows that it encodes 310 putative amino acid residues separated by 4 introns and is designated as fleA. It shows approximately 26% similarity with the gene encoding a fucose-specific lectin of Aleuria aurantia (AAL). The gene was overexpressed under control of the melO promoter in a submerged culture of A. oryzae. The fleA gene product showed hemagglutination activity against rabbit erythrocytes. A hemagglutination inhibition assay of monosaccharides showed that this lectin specifically binds to L-fucose and weakly reacts with mannose and N-acetyl-neuraminic acid.

Amino Acid Sequence↗

Transferable residues from dog fur and plasma cholinesterase inhibition in dogs treated with a flea control dip containing chlorpyrifos.

We studied chlorpyrifos, an insecticide present in a commercial dip for treating ectoparasites in dogs, to estimate the amount of transferable residues that children could obtain from their treated pets. Although the chlorpyrifos dip is no longer supported by the manufacturer, the methodology described herein can help determine transferable residues from other flea control insecticide formulations. Twelve dogs of different breeds and weights were dipped using the recommended guidelines with a commercial, nonprescription chlorpyrifos flea dip for 4 consecutive treatments at 3-week intervals (nonshampoo protocol) and another 12 dogs were dipped with shampooing between dips (shampoo protocol). The samples collected at 4 hr and 7, 14, and 21 days after treatment in the nonshampoo protocol averaged 971, 157, 70, and 26 microg chlorpyrifos, respectively; in the shampoo protocol the samples averaged 459, 49, 15, and 10 microg, respectively. The highest single sample was about 7,000 microg collected at 4 hr. The pretreatment specific activities in the plasma of the dogs were about 75 nmol/min/mg protein for butyrylcholinesterase (BChE), and 9 nmol/min/mg protein for acetylcholinesterase (AChE). BChE was inhibited 50-75% throughout the study, and AChE was inhibited 11-18% in the nonshampoo protocol; inhibition was not as great in the shampoo protocol. There was no correlation (p<or= 0.05) between length of hair and residues measured that would explain the residue differences among dogs. Transferable residues had largely dissipated during the three weeks after treatment, with the largest decrease occurring during the first week. Greater plasma ChE inhibition was observed at 7 days than at 4 hr, probably reflecting the bioactivation of chlorpyrifos to chlorpyrifos-oxon. Plasma cholinesterase activity did not return to control levels during the 3-week period. The differences between the shampoo and nonshampoo protocols were explained by differences in the techniques of the dip applicators.

Acetylcholinesterase↗

Structural modeling of snow flea antifreeze protein.

The glycine-rich antifreeze protein recently discovered in snow fleas exhibits strong freezing point depression activity without significantly changing the melting point of its solution (thermal hysteresis). BLAST searches did not detect any protein with significant similarity in current databases. Based on its circular dichroism spectrum, discontinuities in its tripeptide repeat pattern, and intramolecular disulfide bonding, a detailed theoretical model is proposed for the 6.5-kDa isoform. In the model, the 81-residue protein is organized into a bundle of six short polyproline type II helices connected (with one exception) by proline-containing turns. This structure forms two sheets of three parallel helices, oriented antiparallel to each other. The central helices are particularly rich in glycines that facilitate backbone carbonyl-amide hydrogen bonding to four neighboring helices. The modeled structure has similarities to polyglycine II proposed by Crick and Rich in 1955 and is a close match to the polyproline type II antiparallel sheet structure determined by Traub in 1969 for (Pro-Gly-Gly)(n). Whereas the latter two structures are formed by intermolecular interactions, the snow flea antifreeze is stabilized by intramolecular interactions between the helices facilitated by the regularly spaced turns and disulfide bonds. Like several other antifreeze proteins, this modeled protein is amphipathic with a putative hydrophobic ice-binding face.

Amino Acid Sequence↗

[Notes on fleas (Siphonaptera) of the fox Cerdocyon thous (Canidae) from an endemic area of visceral leishmaniasis in Jacobina, Bahia, Brazil].

From July to September of 1998, 152 fleas were collected from 18 specimens of the fox Cerdocyon thous captured in the visceral leishmaniasis endemic area of Jacobina, State of Bahia, Brazil. The fleas were identified as: 136 Rhopalopsyllus lutzi lutzi, 11 Pulex irritans, 2 Ctenocephalides canis, 1 Ctenocephalides felis felis and 2 Xenopsylla cheopis.

Animals↗

Potential resistance of crape myrtle cultivars to flea beetle (Coleoptera: Chrysomelidae) and Japanese beetle (Coleoptera: Scarabaeidae) damage.

Field and laboratory studies were conducted to identify potential resistance among crape myrtles, Lagerstroemia spp., to Japanese beetle, Popillia japonica Newman and to flea beetles, Altica spp. Damage ratings revealed variation among cultivars in susceptibility to beetle feeding. Cultivars with Lagerstroemia fauriei Koehne in their parentage exhibited the least amount of damage in choice and no-choice experiments, with few exceptions. The data indicate that both beetle species cause more feeding damage on certain cultivars of Lagerstroemia indica L., such as 'Country Red', 'Twilight', and 'Carolina Beauty' than interspecific cultivars with L. fauriei in their parentage, such as 'Natchez', 'Tonto', and 'Muskogee'. When comparing the effect of parentage on all of the major pests of crape myrtle, L. faurei confers resistance to all pests except crape myrtle aphid. No correlation was found between leaf toughness, leaf color, and leaf nutrients in estimating flea beetle cultivar preference. With this information, growers can more effectively target scouting measures to the most susceptible cultivars. and breeders can select plants that will require the fewest chemical inputs.

Animals↗

Pupation site selection of cat fleas (Siphonaptera: Pulicidae) in various carpet types and its influence on insecticide efficacy.

Pupation sites of cat flea, Ctenocephalides felis (Bouché), larvae were determined in three styles of nylon and one style of wool carpet. Nylon saxony carpet had 59.3% of pupae at the top of the pile and 40% at the base of the pile. In nylon contract carpet, 55.2% of pupae were found at the top, 42.6% in the middle, and only 2.2% at the base of the pile. Nylon loop carpet contained 59.2% of pupae at the base, 25.5% in the middle, and 15.3% in the top of the pile. Wool loop carpet had 92.4% at the base and 3.8% both in the middle and top of the pile. Bioassays comparing the control of pupae manually placed at the base of carpets to that in carpets with natural pupation showed that control of pupae in the latter was 39-68% higher. Pupal control after natural pupation was greatest in nylon saxony and nylon contract carpets and lowest in nylon loop and wool loop carpets. Additional studies demonstrated that vacuuming provided the same level of pupal control on nylon saxony carpet as a spray application of permethrin to the carpet surface. Therefore, pupae that survived chemical and mechanical control treatments in nylon saxony carpet probably pupated away from the surface of the pile. Application of permethrin to the base of nylon saxony carpet did not significantly increase control. Future bioassays with cat flea pupae in carpet should be performed after natural pupation and consider carpet make and style.

Animals↗

Toxicity, persistence, and efficacy of spinosad, chlorfenapyr, and thiamethoxam on eggplant when applied against the eggplant flea beetle (Coleoptera: Chrysomelidae).

A laboratory bioassay was developed for determining the toxicity of spinosad, chlorfenapyr, and thiamethoxam against the eggplant flea beetle, Epitrix fuscula Crotch, on eggplant foliage. Four days after initial exposure, LC50 values were 1.99, 2.50, and 0.88 ppm for spinosad, chlorfenapyr, and thiamethoxam, respectively. By dividing the recommended field rate in ppm by the LC50 value, a field toxicity ratio was determined and ranged from 13.5 for spinosad to 73.9 for thiamethoxam. The high ratios suggest that field rates for all three insecticides could likely be reduced. This was supported by field studies in 2000 in which reduced rates of spinosad and thiamethoxam significantly reduced flea beetle numbers on eggplant. Mortality produced by thiamethoxam occurred more quickly than that for the other tested materials as shown with LT50 values of 1.8, 3.0, and 3.6 and days for thiamethoxam, chlorfenapyr, and spinosad, respectively. Persistence studies indicated that while all three of the tested compounds initially produced high levels of mortality, chlorfenapyr and thiamethoxam produced 50% or greater mortality after 6 d. Our data suggest that future management strategies for E. fuscula on eggplant can be successfully altered to meet the changing needs of the producer. Spinosad was recently registered, is effective against the E. fuscula, and offers a viable alternative to carbamate and pyrethroid insecticides. Thiamethoxam and chlorfenapyr offer high levels of toxicity to E. fuscula and upon registration will offer additional effective tools for management.

Animals↗

Effects of multiple mating on female reproductive output in the cat flea (Siphonaptera: Pulicidae).

Multiple mating behavior of female cat fleas, Ctenocephalides felis (Bouché), was confirmed in this study, and its effects on fecundity and fertility were investigated as well. The number of fertile eggs produced by mated females was close to nil within 7 d after removal of males, but it was resumed when females were exposed to males again on day 7. Multiple-mated females displayed significantly higher fecundity (400.3 eggs per female) and fertility (182.8 viable eggs per female) than single-mated females (61.7 and 19.0, respectively) in the 24-d period, suggesting that multiple mating by females is an advantageous strategy for cat fleas. The duration of first mating averaged 63.1 min. The high ratio (55.56%) and short duration (34.0 min) of impotent mating suggested that cryptic female choice may be involved during copulation.

Animals↗

The flea genus Sigmactenus (Siphonaptera: Leptopsyllidae): a new species from timor and new material from New Guinea and the Philippines.

Both sexes of Sigmactenus timorensis n. sp. are described from Rattus tanezumi and Rattus exulans collected in West Timor, Indonesia. The true host presumably is a native murine rat such as the extant, endemic Rattus timorensis or 1 of several extinct, endemic Timorese rats. Analyses of new collections of Sigmactenus cavifrons and Sigmactenus toxopeusi from New Guinea demonstrate that these fleas show considerable morphological variation. We propose that they represent a single species with the name S. toxopeusi having priority. New collections of Sigmactenus werneri from the Philippines expand the known hosts and geographical distribution of this flea.

Animals↗

New Wolbachia endosymbionts from Nearctic and Neotropical fleas (Siphonaptera).

Several species of fleas (Siphonaptera), ectoparasites of mammals and birds, have recently been shown to harbor species of Wolbachia. Here, we extend this data set to 20 more species of Siphonaptera (Rhopalopsyllidae, Stephanocircidae, Pulicidae, Ceratophyllidae, Ctenophthalmidae, Ischnopsyllidae, Leptopsyllidae, and Malacopsyllidae) from sylvatic populations throughout the Nearctic and Neotropical regions. Using polymerase chain reaction, we targeted the Wolbachia 16S ribosomal DNA (rDNA) gene. Applying maximum parsimony- and maximum likelihood-based algorithms, as well as statistical parsimony, we conducted a phylogenetic analysis of Wolbachia 16S rDNA to evaluate its position within the known Wolbachia spp. The analysis recovered the siphonapteran Wolbachia 16S rDNA sequences as a monophyletic group and shows multiple haplotype connections between the Neotropical and Nearctic Wolbachia strains of fleas.

Animals↗