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Flea allergy dermatitis in dogs diagnosed by intradermal skin tests.

Thirty-three dogs were categorised according to their clinical signs of flea allergy dermatitis and reactivity to feeding fleas (Ctenocephalides felis felis). A soluble extract of whole fleas (FS), fractions of this extract separated by chromatography, and a commercially available flea antigen extract were used in intradermal skin tests (IDST) to establish the presence or absence of type I and type IV hypersensitivity. The reactions were measured and the results were analysed using three grading systems commonly reported in the literature. The results of the IDST for the groups of dogs varied according to the grading system used. FS, the most effective of the antigen preparations, identified 94 per cent of dogs which reacted to feeding fleas when a result was considered positive if the mean diameter of the wheal at the antigen injection site exceeded the diameter of the wheal at the negative control site by five mm at 15 and/or 30 minutes after injection.

Animals↗

Relationships between parasite abundance and the taxonomic distance among a parasite's host species: an example with fleas parasitic on small mammals.

Opportunistic parasite species, capable of exploiting several different host species, do not achieve the same abundance on all these hosts. Parasites achieve maximum abundance on their principal host species, and lower abundances on their auxiliary host species. Taxonomic relatedness between the principal and auxiliary host species may determine what abundance a parasite can achieve on its auxiliary hosts, as relatedness should reflect similarities among host species in ecological, physiological and/or immunological characters. We tested this hypothesis with fleas (Siphonaptera) parasitic on small Holarctic mammals. We determined whether the abundance of a flea in its auxiliary hosts decreases with increasing taxonomic distance of these hosts from the principal host. Using data on 106 flea species from 23 regions, for a total of 194 flea-locality combinations, we found consistent support for this relationship, both within and across regions, and even after controlling for the potentially confounding effect of flea phylogeny. These results are most likely explained by a decrease in the efficiency of the parasite's evasive mechanisms against the host's behavioural and immune defences with increasing taxonomic distance from the principal host. Our findings suggest that host switching over evolutionary time may be severely constrained by the coupling of parasite success with the relatedness between new hosts and the original host.

Animals↗

Disposition of 3H-selamectin and 3H-ivermectin in the brain of the cat flea Ctenocephalides felis felis using micro-image analysis.

In addition to intrinsic potency and metabolic stability, the disposition of an antiparasitic drug within the target parasite plays a major role in determining drug activity. A novel technique that allows the disposition of radiolabelled drugs to be visualised within the body of the cat flea (Ctenocephalides felis felis) is described. The concentrations of two macrocyclic lactones, (3)H-selamectin and (3)H-ivermectin, within the supra- and sub-oesophageal ganglia of the flea brain following in vitro feeding of fleas on different doses of drug solubilised in calf blood have been measured. Drug disposition was visualised in cryostat sections of fleas using a micro-image analysis (MIA). A relationship between the concentration of radioactivity in the ganglia and the dose of drug in the blood meal was obtained. The concentration of selamectin in the ganglia was significantly higher than ivermectin at all doses investigated. The enhanced concentration of selamectin, at a site rich in glutamate-gated chloride channels may, in part, explain the higher potency of selamectin against fleas compared to ivermectin.

Animals↗

Identification, cloning, and characterization of a major cat flea salivary allergen (Cte f 1).

An 18 kDa protein isolated from saliva of the cat flea, Ctenocephalides felis, elicits a positive intradermal skin test (IDST) in 100 and 80% of experimental and clinical flea allergic dogs, respectively. Using solid-phase enzyme-linked immuno assay (ELISA), this protein detected IgE in 100 and 80% of experimental and clinical flea allergic dogs, respectively. A cDNA (pFSI) encoding a full-length Cte f 1 protein was isolated from a C. felis salivary gland cDNA library, using a combination of PCR and hybridization screening. This cDNA is 658 bp in length, and contains an open reading frame of 528 bp. The open reading frame encodes a protein of 176 amino acids, consisting of an 18 amino acid signal sequence and a 158 amino acid mature protein. The calculated molecular weight and pI of the mature protein are 18106 Da and 9.3, respectively. The protein, named Cte f 1, is the first novel major allergen described for canine flea allergy. Recombinant Cte f 1 (rCte f 1) was expressed in Escherichia coli, Pichia pastoris and baculovirus infected Trichoplusia ni cells. Approximately, 90% of the rCte f 1 expressed in E. coli accumulated in insoluble inclusion bodies, which could be refolded to a soluble mixture of disulfide isomers with partial IgE binding activity. Small quantities of an apparently correctly refolded form of rCte f 1, which had IgE binding activity equal to the native antigen, was isolated from the soluble fraction of E. coli cells. However, P. pastoris and baculovirus infected insect cells expressed and secreted a fully processed, correctly refolded and fully active form of rCte f 1. Mass spectrometry analysis of the active forms of rCte f 1confirmed that eight intact disulfide bonds were present, matching the number observed in the native allergen. The relative ability of rCte f 1 to bind IgE in the serum of flea allergic animals, produced in these three expression systems, matched that of the native allergen. Competition ELISA demonstrated that approximately 90% of the specific IgE binding to native Cte f 1 could be blocked by the different forms of rCte f 1.

Alkylation↗

Efficacy of selamectin against adult flea infestations (Ctenocephalides felis felis and Ctenocephalides canis) on dogs and cats.

Selamectin was evaluated in eight controlled studies (4 in dogs, 4 in cats) to determine the efficacy of a single topical unit dose providing the recommended minimum dosage of 6mgkg(-1) against Ctenocephalides felis felis and Ctenocephalides canis fleas on dogs and against C. felis on cats. In addition, the effect of bathing on the efficacy of selamectin against C. felis was evaluated. Identical studies were performed in Beagles and domestic shorthaired cats. For each study, animals were allocated randomly to treatments of 8-12 animals each. All studies (dog studies A, B, C, and D and cat studies A, B, C, and D) evaluated the efficacy of selamectin without bathing. In addition, study C in both dogs and cats evaluated efficacy with a shampoo bath at 24h after dosing, and study D evaluated the efficacy of selamectin with water soaking at 2h after dosing or with a shampoo bath at 2-6h after dosing. Dog study B evaluated efficacy against C. canis, whereas all other studies used C. felis. In each study, selamectin was administered on day 0 as a topical dose that was applied directly to the skin in a single spot at the base of the neck in front of the scapulae. Dogs and cats were infested with approximately 100 viable unfed C. felis or C. canis on days 4, 11, 18, and 27. On days 7, 14, 21, and 30, approximately 72h after infestation, a comb count of the number of viable fleas present on each animal was made. For C. felis and C. canis for dogs and cats, compared with controls, selamectin achieved significant reductions in geometric mean adult flea comb counts of > or =98.9% on days 7, 14, and 21 in all eight studies. On day 30, the reduction for C. felis remained at or above 98.0%. This included the dogs and cats that were soaked with water or bathed with shampoo at 2, 6, or 24h after treatment. There were no significant (P>0.05) differences between the flea counts from selamectin-treated animals in these studies, regardless of bathing status. On day 30, a significant reduction of 91.8% was achieved against C. canis on dogs. Thus, these studies demonstrated that a single topical unit dose of selamectin was highly effective against adult fleas on dogs and cats for at least 27 days.

Administration, Topical↗

Cloning and characterization of five cDNAs encoding peritrophin-A domains from the cat flea, Ctenocephalides felis.

Five cDNAs encoding peritrophin-A domains were identified as expressed sequence tags (ESTs) from flea hindgut and Malpighian tubule (HMT) cDNA libraries. The full-length cDNAs for each were subsequently isolated and sequenced. Three of the encoded proteins were similar to published peritrophin sequences, and thus were called "peritrophin-like", or PL1, PL2, and PL3. The other two sequences had similarity to both mucin and peritrophin proteins, and were called "mucin/peritrophin-like", or MPL1 and MPL2. The predicted protein sequences encoded by these cDNAs all contained a signal sequence and one or more peritrophin-A domains, which have been shown in other proteins to bind chitin. Aside from the peritrophin-A domains, the sequences shared little or no similarity to each other or to other proteins in the GenBank non-redundant database. The predicted protein sequences were variable in size, ranging in length from 81 to 453 amino acids. The two MPL proteins contained putative N-linked and O-linked glycosylation sites, including a region of seven nearly perfect tandem repeats in the MPL1 protein sequence. Northern blot analysis of different flea lifestages and fed adult timepoints showed distinct mRNA expression patterns for each gene, although all five transcripts were primarily or exclusively detected in the HMT tissues in adults. The PL1 protein was detected by immuno-blot in soluble and insoluble protein extracts from unfed and fed adult fleas. The PL1 protein from the insoluble fractions appeared to be approximately 1 kDa larger than the PL1 protein from the soluble protein fractions. Immunohistochemistry performed on flea thin sections revealed that the PL1 protein was detected in the Malpighian tubules, hindgut, rectum, and trachea. Unpurified native PL1 protein from both soluble and insoluble protein fractions was tested for chitin-binding activity but did not bind to chitin under the conditions tested. These results show that the flea peritrophin-like proteins may have biological functions that are distinct from the peritrophic matrix and from the binding of chitin.

Amino Acid Sequence↗

Myxomatosis: some observations on breeding the European rabbit flea Spilopsyllus cuniculi (Dale) in an animal house.

Rabbit fleas for use in Myxomatosis investigations have been successfully bred on rabbits in an animal house. The timing of emergence appeared to be governed by a biological timing control interacting with different forms of disturbance. Yield was found to be related to litter size, the time the doe and her kittens were removed from the nest, the number of fleas put onto a doe before littering and the mean ambient temperature to which the doe was exposed in the week pre-partum. The survival rate of fleas in storage was affected by temperature, the degree of crowding, moisture content of the containers, whether fleas were fed or unfed and the source of fleas in terms of emergence times.

Animals↗

Establishment of the cat flea (Ctenocephalides felis felis) on the ferret (Mustela putorius furo) and its control with imidacloprid.

As the ferret, Mustela putorius furo L. (Carnivora: Mustelidae), is becoming increasingly popular as a pet animal and as it is susceptible to the cat-flea, Ctenocephalides felis felis Bouché (Siphonaptera: Pulicidae), an experimental model was established for evaluating insecticidal treatments on this host. A high establishment rate (76.7-91.8%) was recorded when 60 unfed adult C. felis were placed on ferrets. This provided an adequate infestation for chemotherapeutic evaluation without causing undue discomfort to the host. Twelve ferrets were allocated to two groups matched for sex and individual ability to sustain a flea population. One group was treated topically with an imidacloprid spot-on formulation at a dose rate of 10 mg/kg body-weight on Day 0. All ferrets were infested with C. felis on Days -1, 7, 14, 21 and 28, and flea counts were performed 8 and 24 h post-treatment and one day after each subsequent infestation. Fleas were removed at all but the 8 h count (when they were returned to their host). Flea burdens were reduced by 95.3% (P < 0.001) within 8 h of treatment and 100% efficacy was recorded at 24 h. At 1, 2, 3 and 4 weeks post-treatment, protection against re-infestation was 92.9% (P < 0.001), 55.7% (P < 0.02), 18.3% (NS) and 7.4% (NS), respectively. Thus, at this dose rate, imidacloprid gave excellent efficacy against a resident C. felis population and provided a high level of residual activity for at least one week after treatment.

Administration, Topical↗

Palatability and toxicity of fipronil as a systemic insecticide in a bromadiolone rodenticide bait for rat and flea control.

Vector control in plague-infested areas requires a simultaneous killing of rodents and their fleas. We investigated the efficacy of a combination of a systemic insecticide, fipronil, in a rodenticide bait formulation under laboratory conditions. Four different concentrations of fipronil (0.05%, 0.005%, 0.0005% with acetone as a solvent, and 0.05% with propylene glycol as a solvent) and two controls (solvents only) were combined with the rodenticide bait (crushed organically grown wheat with 0.005% bromadiolone). Each concentration was offered together with an untreated non-poisonous challenge bait to 10 singly caged Rattus rattus L., each with 100 rat fleas Xenopsylla cheopis Rothschild (Siphonaptera: Pulicidae) in the nest. Treated bait consumption was relatively low and an unsatisfactory rat mortality of around 50% only was obtained in all tests. The palatability of the bait, however, was not affected by the fipronil concentration. Even at the lowest fipronil concentration, average flea mortality was still above 95%, and doses of more than I mg fipronil per kg rat body weight gave a nearly complete kill of fleas. Fipronil can be highly effective as a systemic insecticide to for flea control, provided that a more attractive bait base for roof rats is used.

4-Hydroxycoumarins↗

Abiotic factors influencing the ecology of wild rabbit fleas in north-eastern Spain.

During 1992, the population dynamics of rabbit fleas were compared at two sites in north-eastern Spain. The sites differed mainly in terms of annual rainfall and soil type. All flea species showed seasonal cycles of abundance, although peaks in numbers occurred at different times, reflecting their specific adaptations for coping with climatic variables. Adult Spilopsyllus cuniculi (Dale) (Siphonaptera: Pulicidae) were found largely parasitizing rabbits in spring and adult Caenopsylla laptevi (Beaucournu etal.) (Siphonaptera: Ceratophyllidae) in the autumn. In contrast, monthly flea indices of Xenopsylla cunicularis (Smit) (Siphonaptera: Pulicidae) and Echidnophaga iberica (Ribeiro et al.) (Siphonaptera: Pulicidae) peaked in summer. Spilopsyllus cuniculi was present at both sites, but was less common on the drier site, where monthly mean temperature and annual rainfall approached the flea's physiological limits. By contrast, E. iberica, X. cunicularis and C. laptevi, known to be better adapted for dryness, showed the opposite patterns of abundance. Nevertheless, even these arid-adapted species took advantage of the milder and wetter spring (X. cunicularis and E. iberica) or autumn (C. laptevi) for breeding and larval development. Although environmental temperature, rainfall and soil texture will influence the microclimate of the burrows where the flea larvae develop, burrow humidity seems to be more dependent on soil characteristics and past rainfall rather than the humidity of the external air.

Animals↗

Annual cycles of four flea species in the central Negev desert.

Bionomics of fleas (Siphonaptera) parasitizing rodent hosts, mostly the gerbil Gerbillus dasyurus (Wagner) and the jird Meriones crassus Sundevall (Gerbillidae), were investigated in the central Negev desert of Israel. Populations were sampled weekly (by Sherman trapping of hosts) from August 2000 to July 2001. Among 1055 fleas of nine species captured, four species predominated (94%). Two species of Pulicidae, Xenopsylla dipodilli Smit and X. ramesis (Rothschild), reproduced perennially, whereas adults of Nosopsyllus iranus theodori Smit (Ceratophyllidae) and Stenoponia tripectinata medialis Jordan (Hystrichopsyllidae) occurred only in cool months (October-March). During their main activity season on the most infested host species (estimated from > 300 trap-nights/month), prevalence of these four flea species reached 40-70%, 20-30%, 100% and 50-70%, respectively, with infestation intensities of 2-2.7, 7-12, 2-3.5 and 2.5-7 fleas per infested rodent, respectively. Xenopsylla dipodilli oviposition peaked during autumn with parous rate > 80% by September-October. During December-April, the majority of X. dipodilli females were immature and/or nulliparous (defined as having mature ovaries but no follicular relics). In contrast, X. ramesis had two reproductive peaks, in mid-spring and autumn, evidenced by the influx of immature females in late spring and summer (30-40%) and in winter (20-30%) after maximal parous rates: 80-100% in March-April and 95-100% in October-November. Nosopsyllus iranus theodori and Stenoponia tripectinata medialis adults occurred only during cool months. At the beginning of activity, during October-November, the sex ratio of N. i. theodori was strongly biased to females (86%) that were immature and/or nulliparous. In winter, adult females were 52-65% parous and 10-32% immature. In March, as the adult population of N. i. theodori declined, 78% of females were parous and 12% immature. Seasonal activity of S. t. medialis (November-March) was shorter than for the other three species; females were predominantly nulliparous in November (80%), after which the proportion of parous females increased gradually to 84% in February. Two females of S. t. medialis collected in March were mature but nulliparous, suggesting that this species of flea might 'oversummer' (as pupae or teneral adults) in the cocooned stage.

Animals↗

Classic flea-borne transmission does not drive plague epizootics in prairie dogs.

We lack a clear understanding of the enzootic maintenance of the bacterium (Yersinia pestis) that causes plague and the sporadic epizootics that occur in its natural rodent hosts. A key to elucidating these epidemiological dynamics is determining the dominant transmission routes of plague. Plague can be acquired from the bites of infectious fleas (which is generally considered to occur via a blocked flea vector), inhalation of infectious respiratory droplets, or contact with a short-term infectious reservoir. We present results from a plague modeling approach that includes transmission from all three sources of infection simultaneously and uses sensitivity analysis to determine their relative importance. Our model is completely parameterized by using data from the literature and our own field studies of plague in the black-tailed prairie dog (Cynomys ludovicianus). Results of the model are qualitatively and quantitatively consistent with independent data from our field sites. Although infectious fleas might be an important source of infection and transmission via blocked fleas is a dominant paradigm in the literature, our model clearly predicts that this form of transmission cannot drive epizootics in prairie dogs. Rather, a short-term reservoir is required for epizootic dynamics. Several short-term reservoirs have the potential to affect the prairie dog system. Our model predictions of the residence time of the short-term reservoir suggest that other small mammals, infectious prairie dog carcasses, fleas that transmit plague without blockage of the digestive tract, or some combination of these three are the most likely of the candidate infectious reservoirs.

Animals↗

Blood consumption by the cat flea, Ctenocephalides felis (Siphonaptera: Pulicidae).

The volume of blood consumed by actively reproducing female cat fleas, Ctenocephalides felis (Bouché), while feeding in confinement feeding chambers on cats for 3 h, was investigated using two radionuclide blood tags (51Cr-erythrocyte and 125I-albumin) and the gravimetric method. Female fleas consumed an average of 0.110 ml (+/- 0.026 ml) of blood per 100 females in 3 h as determined using the dual radionuclide system. The single 51Cr-erythrocyte tag overestimated blood consumption by 11.3% and the single 125I-albumin tag underestimated blood consumption by 6.4%. The gravimetric method underestimated blood consumption by 72.2% compared with the dual radionuclide value. Investigations of blood consumption of nonconfined female fleas were conducted with cats housed in metabolic cages and restricted from grooming. These investigations were conducted using the single 51Cr-erythrocyte tag, and the data obtained were corrected to compensate for using a single erythrocyte tag. Female cat fleas consumed an average of 13.6 microliters (+/- 2.7 microliters) of blood per day, which was equivalent to 15.15 times their body weight. In an additional study, direct exposure to 40 kHz ultrasound did not reduce blood consumption by the fleas.

Animals↗

Acquisition of the cat scratch disease agent Bartonella henselae by cat fleas (Siphonaptera:Pulicidae).

We assayed the ability of cat fleas to become infected with Bartonella henselae, using an artificial feeding device. Fleas fed a concentration of 1 x 10(5) cfu/ml in blood were examined using immunofluorescent antibody assay and polymerase chain reaction. Bacteria were present in the gut at 3 h, and persisted up to 9 d after infection. Qualitatively, the density of B. henselae was greater in the flea gut at 9 d, indicating that replication was occurring in the gut. B. henselae also was detected in the feces of infected fleas 9 d after infection, and produced viable colonies upon inoculation onto heart infusion agar/rabbit blood plates. Our results indicate that fleas can maintain infection with B. henselae, and may play a role in the transmission of this bacterium from infected cats to humans.

Animals↗

Toxicity and histopathology of the growth regulator pyriproxyfen to adults and eggs of the cat flea (Siphonaptera:Pulicidae).

Adult cat fleas, Ctenocephalides felis (Bouché), exposed continuously to pyriproxyfen died within 8-10 d. Microscopic examination of 7-d-old adults indicated death was caused by histopathological damage to fat body, Malpighian tubules, midgut epithelia, salivary gland cells, and other internal tissues. Fleas were killed by pyriproxyfen regardless of whether they were held as unfed adults on treated filter paper or as feeding adults on treated dog hair. In addition to these toxic effects on adults, pyriproxyfen also induced formation of large autophagic vacuoles in maturing oocytes leading to partial reabsorption of yolk, degeneration of the nucleus, and lysis of the follicular epithelium. Consequently, most of the eggs laid by treated fleas lacked a nucleus, had a poorly formed chorion, and were ruptured during ovulation. These laboratory results suggest that continuous exposure of fleas to pyriproxyfen on a host animal could prevent deposition of viable eggs and eventually kill adults, thereby controlling all stages of flea development.

Animals↗

Factors affecting the seasonal abundance of ground squirrel and wood rat fleas (Siphonaptera) in San Diego County, California.

Abiotic and biotic factors affecting the seasonal abundance of 3 species of sylvatic fleas on their hosts were examined at 7 sites in San Diego County, California. Indices for Oropsylla (Diamanus) montana (Baker) were usually highest on Spermophilus beecheyi nudipes (Huey) when < 18.4 degrees C (October-December), whereas those for Hoplopsyllus anomalus (Baker) were highest when > 18.4 degrees C (July-September). O. montana was affected most by ambient temperature (inversely for the coastal site [134 m] and directly for most mountain sites [> 1,183 m]). O. montana was affected most (directly) by relative humidity at the inland valley site, which concurs with this flea being most abundant in other areas during periods yielding higher relative humidties and moderate ambient temperatures. H. anomalus was influenced most (directly) by ambient temperature at lower sites (< 1,183 m) and by host activity at higher ones (> 1,456 m). Usually nonrandom distribution findings, when indices for each squirrel flea were higher on some hosts, indicate that flea numbers are correspondingly higher in some nests and burrows because of more favorable microconditions. Data also indicated that plague may persist at higher sites in southern California or in other areas where climatic conditions permit increased and prolonged interactions between more abundant O. montana and its host during spring through summer. Whereas, plague may have less chance of persisting at lower sites because these interactions are decreased and shortened during the warmer months. Squirrels may become infected with plague following hibernation and again when reoccupying colonial burrows. The flea Orchopeas sexdentatus was affected most (inversely) by relative humidity at a coastal site, with higher indices occurring on fewer Neotoma lepida Thomas and on more abundant Neotoma fuscipes macrotis Thomas during colder months. Such increased flea/Neotoma fuscipes Baird activity at lower sites may favor plague amplification during the winter which may involve ground squirrels and other rodents in the spring.

Animals↗

Use of orally administered chitin inhibitor (lufenuron) to control flea vectors of plague on ground squirrels in California.

The efficacy of orally administered lufenuron, a chitin inhibitor, to control fleas on California ground squirrels, Spermophilus beecheyi (Richardson), was evaluated during a 2-yr study in Santa Barbara County, CA. Results demonstrated that use of a host-targeted feed cube containing lufenuron was effective in significantly reducing the burden of Oropsylla montana (Baker) and Hoplopsyllus anomalus (Baker) fleas on ground squirrels. A flea index that indicated a mean number of fleas per squirrel of 10.0 decreased to 1.3 after 2 treatments in season 1, and to 0.7 and 0.2 after the 3rd and 4th treatments, respectively, in season 2. A cost comparison of this new method compared with a traditional reactive, emergency, insecticide-based plague control program demonstrated a cost reduction of approximately 90%. The results of this study indicated that a lufenuron feed cube was an effective, cost-saving, and proactive technique for controlling fleas on California ground squirrels, and thus reducing the risk of disease transmission in plague endemic regions.

Administration, Oral↗

Seasonal patterns of rodents, fleas and plague status in the Western Usambara Mountains, Tanzania.

Field and commensal rodents were live-trapped at three villages in an active focus of plague (Yersinia pseudotuberculosis pestis) in Lushoto District, Western Usambara Mountains, Tanga Region, Tanzania, from December 1983 to November 1984. Their flea ectoparasites were collected, identified and counted. The rodent carcasses were serologically examined for specific plague antibodies and antigens, and bacteriologically examined for bipolar staining bacilli. A total of 1758 traps were set during the 12-month period and 924 animals were caught. From these, 1037 fleas were collected. Rattus rattus (L.), Praomys natalensis (Smith) and Lophuromys flavopunctatus Thomas comprised the largest proportions of the rodent population, while Dinopsyllus lypusus Jordan & Rothschild, Ctenophthalmus calceatus Waterston and Xenopsylla brasiliensis (Baker) were the dominant flea species. Rodents were most abundantly trapped during December and January. Flea indices were highest from December to May. Human plague was most active from November to March. Rodents contained plague antibodies every month except May and July, with a peak in September. Plague antigens and bipolar bacilli were detected in rodent organs during January-April. From the product of abundance and infection rate, the most prevalent rodent hosts of plague appeared to be R. rattus, Otomys angoniensis Wroughton, P. natalensis and Pelomys fallax (Peters). Continuous integrated control of rodents and fleas was recommended, reinforced by quarantine and maintenance of a surveillance service for clinical detection, diagnosis and treatment of patients in the plague endemic area.

Animals↗