Effects of ivermectin (MK-933) on the reproductive rate of Aedes aegypti (Diptera: Culicidae)

Aedes aegypti (L.) females were blood fed a single time on rabbits previously injected subcutaneously with ivermectin at 10 or 50 times the labeled dose recommended for cattle (0.2 mg [AI]/kg body weight). Although adult mosquito survival was unchanged by the low dose, fecundity and hatch were depressed slightly. Females fed on rabbits at the high dose exhibited reduced survival and egg production compared with females fed on control rabbits. Eggs from these females also were less likely to hatch, and subsequent larval survival was lower than controls. The effect of ivermectin on these reproductive parameters diminished as the time between drug administration and blood meal increased.     

Distribution and pharmacokinetics of precocene-II in the tick Ornithodoros parkeri (Acari: Argasidae)

Injected [3H]precocene-II spread from the hemocoel of Ornithodoros parkeri (Cooley) ticks to gut, salivary glands, synganglion, ovary, and the remainder of the body (carcass). Analyses at 2, 24, and 48 h after injection of precocene revealed that radioactivity remained at low levels in the synganglia and ovaries of fed-mated and fed-virgin females. Radioactivity increased gradually during 48 h after injection in salivary glands of fed-mated and fed-virgin ticks. It also increased gradually in the gut of fed virgins but not in fed-mated females. The carcasses contained the majority of total radioactivity at 2, 24, and 48 h after injection. Most of the radioactivity was found in ether-soluble compounds and only minimal amounts in water-soluble metabolites. Analysis of tick excreta and washings indicate that female O. parkeri excrete injected precocene very slowly; only 6% of total radioactivity was recovered in these samples during the 48 h after injection. Thin-layer chromatographic analyses of ether extracts of salivary glands and carcasses from fed-mated ticks 24 h after injection showed that the majority of the injected [3H]precocene-II was present in unchanged form.     

Effect of ivermectin on the volume of blood ingested by two species of ticks (Acari: Ixodidae) feeding on cattle

Females of the lone star tick, Amblyomma americanum (L.), and the American dog tick, Dermacentor variabilis (Say), were fed on ivermectin (Ivomec)-treated and untreated bovines to determine the effect of the acaricide on volume of blood ingested and to compare the weight differences between the treatment and control groups at various time intervals after attachment. Adult females from each genus were collected from Bos tarus hosts and subjected to hematin assays on three collection dates to estimate the volume of blood ingested. Before feeding, lone star ticks contained an average of 2.0 microliters of blood and had an average weight of 5.2 mg. Unengorged American dog tick females had an average blood volume of 3.3 microliters and a mean weight of 5.8 mg. Ticks of both species reacted to ivermectin by expressing lower mean weights, and they consumed smaller quantities of blood. Lone star tick females were significantly affected in terms of amount of blood consumed and body weight changes when compared with control ticks. After feeding on treated cattle, lone star tick females contained smaller quantities of blood than pretreatment females, but there were no significant differences observed until day 12 between the control and the treated groups. American dog tick females on treated hosts had measurable quantities of blood that were significantly different among the experimental groups.     

Attempt to control ticks (Acari: Ixodidae) on deer on an isolated island using ivermectin-treated corn

We report an attempt by an offshore island community to control the vector tick of Lyme disease by providing ivermectin-treated corn to an isolated herd of free-ranging white-tailed deer, Odocoileus virginianus Zimmerman. Medicated corn was supplied in troughs within the island village and from automatic feeders at remote sites during 5 consecutive fall and spring adult tick questing seasons. Acaricide consumption was monitored by assaying its presence in fresh deer pellets and its concentration in deer sera. Its effectiveness was evaluated by recording the number of adult ticks collected from the hides of deer, the number of females becoming sufficiently engorged to oviposit, and the success of subsequent oviposition and eclosion. Entomologic risk was monitored by collecting immature ticks from hosts and adult ticks from vegetation. Estimates based on a subsequent deer reduction program indicated that up to twice as many deer had been present during the project as originally presumed. For this and other reasons related to deer behavior, target levels of serum ivermectin were achieved in a minority of deer. Nevertheless, > 90% control of female tick infestation, subsequent oviposition, and larval eclosion was obtained in those 8 of 16 sampled deer with serum ivermectin levels of > or = 15 ng/ml. In addition, the ratio of females to males, the numbers of females engorging > 10 mg body weight, and the numbers of those eventually hatching, were all significantly less among ticks from island deer in comparison with ticks from untreated deer. No consistent changes in the numbers of ticks found on immature-stage hosts or removed from vegetation were noted within 3 yr of the cessation of treatment.     

Genetic characteristics of Borrelia coriaceae isolates from the soft tick Ornithodoros coriaceus (Acari: Argasidae)

Two Borrelia isolates (CA434 and CA435) cultured from the soft tick Ornithodoros coriaceus were analyzed by contour-clamped homogeneous electric field gel electrophoresis of unrestricted and ApaI-restricted DNA, standard electrophoresis of BamHI- and HindIII-restricted DNA, Southern hybridization, restriction fragment length polymorphism and sequencing of the 16S rRNA gene, and amplification of the 5S-23S intergenic spacer region. These isolates were compared with Borrelia coriaceae type strain Co53, B. burgdorferi sensu stricto strain CA4, and the relapsing-fever spirochete B. parkeri (undesignated). The 16S rRNA region of CA434 and CA435 differed from that of B. coriaceae type strain Co53 by the presence of 1 base (C) at position 367 (GenBank accession no. U42286). The linear plasmid profile of CA434 was similar to that of Co53, and the ApaI, BamHI, and HindIII restriction fingerprints of the total cellular DNA of CA434 and Co53 were similar. In contrast, CA435 differed somewhat from CA434 and Co53, which demonstrates that B. coriaceae is genetically diverse. Southern hybridization showed that the DNAs of CA434 and CA435 hybridized strongly with the digoxigenin-labeled DNA of Co53. Low homology was found between the DNA of Co53 and that of B. parkeri. The 16S rRNA sequence of B. parkeri was identical to previously published results for B. parkeri strain M3001 (GenBank accession number U42296). CA434 and CA435 represent only the second and third isolates of B. coriaceae obtained from any source since its initial isolation from an O. coriaceus tick in 1985. All three B. coriaceae isolates were derived from adult ticks collected from the same locality in northwestern California. Difficulties encountered in detecting B. coriaceae in, and isolating this spirochete from, the tissues of O. coriaceus are discussed. The lack of concordance between different detection or isolation methods suggests that reliance upon a single technique may grossly underestimate the true prevalence of spirochetal infection in wild-caught O. coriaceus ticks.     

Rickettsial infection in ticks (Acari: Ixodidae) collected on birds in southern Brazil

The aim of the study was to evaluate rickettsial infection in ticks from wild birds of the Semidecidual and Atlantic Rainforest remnants of three municipalities of the State of Paraná, southern Brazil. Overall, 53 larvae and nymphs collected from birds were checked for the presence of Rickettsia DNA by molecular tests. Five tick species were tested: Amblyomma aureolatum (Pallas), Amblyomma calcaratum Neumann, Amblyomma longirostre (Koch), Amblyomma ovale Koch, and Amblyomma parkeri Fonseca and Arag?o. A. longirostre ticks were infected with the spotted fever group agents Rickettsia amblyommii strain AL (32.3% infection rate) and Rickettsia parkeri strain NOD (5.9% infection rate). A new rickettsial genotype was detected in the tick A. parkeri (50% infection rate), which had never been reported to be infected by rickettsiae. Through phylogenetic analysis, this new genotype, here designated as strain ApPR, grouped in a cluster composed by different strains of Rickettsia africae, Rickettsia sibirica, and R. parkeri. We consider strain ApPR to be a new genotype of R. parkeri. This study reports for the first time rickettsial infection in ticks from birds in southern Brazil. The role of migrating birds in the dispersal of these rickettsial strains should be considered in ecological studies of spotted fever group agents in Brazil.     

Host specificity of Laelaps (Acari: Laelapidae) in central Brazil.

A study of associations between small mammals and ectoparasites in two adjacent nature reserves near Brasilia, Federal District, Brazil, revealed that ectoparasitic mites of the genus Laelaps Koch are host specific. Data on the prevalence and intensity of mite infestations were combined with measures of the reproductive activity of mite populations to estimate the association of each host-mite pair. Twelve morphologically distinct species of Laelaps occurred in monoxenous and oligoxenous associations with the cricetid rodent genera Oryzomys, Nectomys, Rhipidomys, and Calomys. Nine were referred to species recognized at the time of the study; three were described as new species in a recent publication. When mites initially assigned as the same species infested more than one host species, an examination of the morphological variation within and among mite specimens supported the hypothesis that mites infesting different hosts were reproductively isolated populations. Dispersal pathways apparently were restricted, occurring primarily between conspecific individuals. In one instance, two morphologically similar groups of Laelaps paulistanensis Fonseca were associated with two closely related rodent species, Oryzomys fornesi Massoia and O. nigripes (Olfers). Although the data presented here indicate that these two groups of mites are conspecific, further study is warranted.     

Polymyxin E-1 (colistin sulphate) (neuro-)intoxication in young ostriches ( Struthio camelus spp.)

Colistin (polymyxin E) is a cyclic polypeptide with a potent bactericidal action against most gramnegative bacilli. When used parenterally, polymyxins should be given with great care as they have a very small safety range, and easily induce neurotoxicity and nephrotoxicity. A dose of 39.5mg/kg body weight colistin sulphate injected subcutaneously induced rapid (within 1 to 3 h) mortality in young ostriches. Clinical signs of apathy, lethargy and hypotonia indicative of neurotoxicity of the compound were observed. At postmortem, vascular congestion of brain vessels was seen while, on histology, severe acute oedema was present in the epicardium and the intestinal serosa. Congestion of villi, swelling and vacuolization of the plexus ofAuerbach, as well as intermuscular and perivascular oedema of the heart, were also observed. In view of our observations in ostriches and in other species studied, a dose of >5mg/kg body weight polymyxin E is not considered safe for parenteral administration in ostriches.     

Suitability of white-tailed deer as hosts for cattle fever ticks (Acari: Ixodidae)

White-tailed deer (Odocoileus virginianus L.) were evaluated as an alternate host for the cattle fever tick, Boophilus annulatus (Say). The total of adult female ticks recovered and associated reproductive parameters were compared between deer and bovine hosts. Significantly fewer ticks were recovered from deer than from cattle; this was attributed to the grooming behavior of the deer. The mean weight of ticks taken from deer (243.5 mg) was significantly less than that of ticks recovered from cattle (344.5 mg) with about 15% of the ticks from deer prematurely detached. Egg mass weight from the ticks recovered from deer (129.4 mg) was significantly less than that of ticks from cattle (172.1 mg). A significant difference was observed between the percentage hatch of the eggs of ticks from deer (63.1%) and ticks from cattle (76.7%). The index of reproduction was about 20 times greater for the ticks from the cattle than for the ticks from the deer. When the larval progeny of the ticks recovered from deer were reared on cattle, significantly fewer adult females were recovered than when larvae of ticks recovered from cattle were reared on cattle. This indicates a possible reduction in viability. All other parameters were comparable between the two groups. Thus, deer are biologically suitable hosts for B. annulatus, but significantly fewer ticks complete engorgement and those that complete engorgement have reduced fecundity.     

Host-dependent differences in feeding and reproduction of Ixodes dammini (Acari: Ixodidae)

The frequencies with which adult Ixodes dammini Spielman, Clifford, Piesman and Corwin feed upon white-tailed deer (Odocoileus virginianus), raccoons (Procyon lotor), opossums (Didelphis virginiana), and domestic cats were compared in a wooded site in northeastern United States. By combining estimates of the quantity of ticks feeding on each host species with host densities, we derived the relative contribution made by each kind of host to the feeding of the tick population. To compare engorgement success and reproductive efficacy, we weighed ticks found on these hosts and compared tick weights with the quantity of eggs produced. Deer harbored the most female ticks per individual and, although deer were less abundant than raccoons, they harbored 94.6% of the feeding tick population. Raccoons, cats, and opossums, respectively, provided blood meals to 3.6, 1.6, and 0.2% of feeding adult ticks. Ticks engorged more successfully on cats and raccoons than on deer, and egg production correlated linearly with weight of female ticks collected from both cats and deer. Although these ticks engorged better on other hosts, the vast majority of eggs resulted from ticks that had fed on deer. We conclude that, in this site, the abundance of deer largely determines the abundance of I. dammini, the vector of the agent of Lyme disease.     

Dynamics of the antibody response in cattle infested by Hypoderma (Diptera: Oestridae) after a treatment with injectable and pour-on ivermectin

The dynamics of the anti-Hypoderma antibody response after a treatment with injectable and pour-on ivermectin against first instars of Hypoderma sp. were determined in naturally infested cows. Cattle were randomly assigned to three groups: group 1, which served as an untreated control; group 2, treated with pour-on ivermectin at a dose of 500 microg/kg of body weight, and group 3, which received 200 microg/kg of body weight of ivermectin by subcutaneous injection. Both ivermectins showed total efficacy against L-1 of Hypoderma sp. In untreated control animals, grubs became detectable in the 2 mo after treatment. Indirect enzyme-linked immunosorbent assay (ELISA) showed a significant increase in group 3 antibody levels 1 mo after treatment, whereas in group 2 the increase was not significant. Following the peak of anitibody activity in group 3, antibody levels declined at a faster rate than in those treated topically. In both treated groups, cattle remained serologically positive for 4 mo after ivermectin treatment. Thus, serological surveillance programs for cattle grubs conducted during this posttreatment period will not be influenced by treatment with ivermectin. The persistence of antibody levels after larval destruction makes the indirect ELISA unsuitable for the prediction of the efficacy of treatment.     

Tick (Acari) infestations of bats in New Mexico

A total of 278 bats belonging to 16 species was examined for ticks from various sites in New Mexico from 1994 to 1998. Seven species of bats were parasitized by ticks: larvae of Ornithodoros kelleyi Cooley & Kohls, Ornithodoros rossi Kohls, Sonenshine & Clifford (Argasidae), or both. Both species of ticks are reported from New Mexico for the first time. Infestation prevalences for parasitized bats ranged from 2 to 25% on different host species for O. kelleyi and from 7 to 25% for O. rossi. The pallid bat, Antrozous pallidus, and the big brown bat. Eptesicus fuscus, were parasitized by both tick species. No distinct host specificity was noted for either tick species.     

Deer density and the abundance of Ixodes scapularis (Acari: Ixodidae)

The abundance of Ixodes scapularis Say (Ixodes dammini Spielman, Clifford, Piesman & Corwin), the vector tick of the Lyme disease spirochete and other human pathogens, is related to the presence of its primary reproductive stage host, white-tailed deer (Odocoileus virginianus Zimmerman). However, this relationship has not been quantified in terms that would guide wildlife management in areas in which the public is, or is likely to become, exposed to infected ticks. In this study, deer density and tick abundance were measured in an emergent area for Lyme disease at three spatial scales using estimation methods appropriate for each. Simple linear regression was used to relate (1) the number of ticks found on deer at tagging stations in southern Maine to harvest-derived estimates of the density of deer within the towns in which they were killed, (2) tick densities estimated from fall flagging counts to deer densities estimated from pellet group counts made within multiple transects distributed through 5.2-km2 study sites, and (3) tick counts to pellet group counts within the individual transects. At the broadest scale, ticks on deer decreased with elevation and distance from the coast and increased with deer density, although deer and tick presence were only weakly related. Among the 5.2-km2 study sites and within individual transects, tick abundance related more strongly to deer pellet group counts. Few ticks were collected at deer densities <7/km2.     

Improved antifilarial activity of ivermectin in chitosan–alginate nanoparticles against human lymphatic filarial parasite, Brugia malayi

The current antifilarial treatments are not up to the mark partly due to deep location of filarial parasites in the human lymphatic system. We report here on the improvement in the antifilarial activity of ivermectin (IVM) using chitosan–alginate nanoparticles prepared by modified complex coacervation method. The nanoparticles were spherical having 155 nm size and 4.56 and 75.67 % loading and entrapment efficiency respectively for IVM. The delivery system maintained the sustained release and significantly augmented the microfilaricidal (MIF) activity at a single low dose (200 μg/kg body weight, subcutaneously) in contrast to much higher dose of free ivermectin (400 μg/kg body weight, subcutaneously) against human lymphatic filariid, Brugia malayi in rodent host, Mastomys coucha. To substantiate increase in MIF activity, pharmacokinetics study was designed on Wistar rats which revealed a greater peak plasma concentration (45.3?±?1.79 ng/ml), area under the concentration curve (298?±?38.7 ng d/ml) and extended mean residence time (23.4?±?8.56 days)of IVM in chitosan–alginate nanoparticles. Administration of 25 mg/kg of diethylcarbamazine following nanoparticle therapy significantly improved the MIF and macrofilaricidal action of encapsulated drug and was considered superior in this study.     

Evaluation of the Toxicological Profile of the Leaves and Young Twigs of Caesalpinia Bonduc (Linn) Roxb

Acute and sub-acute toxicological effects of ethanolic extract of the leaves and young twigs of Caesalpinia bonduc were carried out on albino rats. Single extract doses from 2000 to 5000 mg/kg body weight were administered orally and monitored for 14 days in acute study, while extract doses from 200 to 1600 mg/kg body weight were orally administered daily for 28 days in sub-acute study and recovery was assessed 14 days after dosing. Biochemical, haematological and histopathological examinations were carried out. There was no mortality in the experimental animals in all acute treatment doses. However, there were significant alterations in the biomarkers and induced cellular damage to the liver in all acute treatment doses. In the sub-acute toxicity treatment, the assessed biomarkers were unaffected at extract dose of 200 mg/kg body weight compared to control, while significant changes were observed in rats administered with extract doses of 400 mg/kg body weight and above. No significant difference was observed between the tested groups and the recovery groups in the sub-acute toxicity study. In conclusion, the ethanolic extract of C. bonduc could be toxic to selected organs of the rat body in acute and sub-acute treatments.     

Mechanical transmission of Venezuelan equine encephalomyelitis virus by hematophagous mites (Acari).

Studies were undertaken to determine whether two species of hematophagous mites could transmit Venezuelan equine encephalomyelitis (VEE) virus to suckling mice. The chicken mite, Dermanyssus gallinae (De Geer), mechanically transmitted this virus to previously uninfected mice up to 16 h after taking a blood meal from a viremic animal. In contrast, Laelaps kochi Oudemans failed to transmit virus to mice in any feeding trials. Virus did not replicate in either species of mite but was detected in 20% of D. gallinae and in 12% of L. kochi specimens tested 48 h after a viremic blood meal. In mites inoculated intracoelomically with virus, recovery rates declined over time, but virus was detected up to 7 d later in 5% of D. gallinae and up to 3 d later in 10% of L. kochi. Biological transmission of VEE virus was not demonstrated in either species of mite.     

Immune responses of interferon gamma (IFN-gamma) knock out mice to repeated Haemaphysalis longicornis (Acari: Ixodidae) nymph infestations

To investigate the immunological mechanisms of acquired resistance to tick infestation, interferon gamma (IFN-gamma) deficient mice (IFN-gamma mice) were used to assess interleukin-4 (IL-4) and antibody production levels against tick salivary gland antigen on three successive infestations with Haemoaphysalis longicornis Neumann nymphs. The engorged body weight of the ticks decreased during the second and third infestations. Similar observations were noted in IFN-gamma+/+ mice. However, the engorged body weight of the ticks from IFN-gamma +/+ mice were considerably lower than those from IFN-gamma-/- mice. A marked increase in antibody production during the second and third infestations was observed indicating that IFN-gamma-/- mice could acquire immunological resistance against H. longicornis nymphs. Moreover, IL-4 levels were higher during the first and third infestations but decreased during the second infestation. IL-4 levels were significantly higher in IFN-gamma-/- mice than in IFN-gamma+/+ mice. We have shown here that the statistically significant high IL-4 levels observed in IFN-gamma-/- mice may be a result of type 2 helper cell (Th2) polarization. However, the apparently higher IL-4 levels during the first and third infestations and the notable decline during the second infestation suggest that other cytokines or factors in the host immune system may play a part in regulating IL-4 levels.     

Endotoxin hepatotoxicity augmented by ethanol

To determine whether alcohol increases endotoxin hepatotoxicity, we administered ethanol (4.8 g/kg body wt in 4 ml of water) to rats through a gastric tube, then immediately injected endotoxin (2, 2.5, or 3 mg/kg body wt). In the rats pretreated with ethanol, the injection of 2 mg/kg body wt of endotoxin induced a slight rise of serum transaminase. However, when 2.5 mg/kg body wt of endotoxin was given, there were no significant histopathological or biochemical differences between the rats pretreated with ethanol and those pretreated with water. Moreover, there was no significant difference in mortality rates between the rats pretreated with ethanol and the controls when 3 mg/kg body wt (LD50) of endotoxin was injected. These results suggest that acute administration of alcohol enhances endotoxin hepatotoxicity when the dose of endotoxin is small, but that the effect of alcohol is masked when larger doses of endotoxin are given.     

Quantitative studies of host immunoglobulin G in the hemolymph of ticks (Acari)

A radioimmunoassay was used to measure the concentration of host immunoglobulin G (IgG) in the hemolymph of female hard and soft ticks. Hyalomma excavatum Koch, Rhipicephalus sanguineus Latreille, Ornithodoros tholozani (Laboulbene and Megnin), and O. moubata (Murry) were fed on rabbits immunized with ovalbumin; Argas persicus (Oken) was fed on chickens immunized with cytochrome 'C.' At 24 h after feeding, the concentration of antiovalbumin IgG in the hemolymph was 7 micrograms/ml for H. excavatum, 5 micrograms/ml for R. sanguineus, and 0.15 micrograms/ml for O. moubata; the percentage of intact IgG molecules was 30, 44, and 100%, respectively. Host IgG was not detected in the hemolymph of O. tholozani and A. persicus. There was no increase in the concentrations of host IgG in the hemolymph of the soft ticks during the first week following the blood meal. The potential contribution to the resistance of hosts against ticks by host antibodies that cross into the tick hemocoel is discussed.     

Studies on the Chemoprophylaxis of subperiodicBrugia malayi infection in the leaf monkey (Presbytis melalophos) with diethylcarbamazine citrate

The chemoprophylactic use of diethylcarbamazine citrate at total oral doses of 15–180 mg/kg body weight was tested against subperiodicBrugia malayi infection in the leaf monkey (Presbytis melalophos). A total dose of 45 mg/kg body weight given over 9 days killed all developing over 7 days killed all fourth stage larvae. The minimum effective dose that prevents infection would be 5 mg/kg body weight daily for 7 days every month.