Borreliae in Ixodes ricinus ticks feeding on humans

A total of 298 Ixodes ricinus (L.) ticks (Acari: Ixodidae) feeding on humans in the Czech Republic were tested for borreliae (Borrelia burgdorferi sensu lato) by darkfield microscopy between 1997 and 2003. A majority (68%) of the supplied I. ricinus ticks were nymphs, 25% were females and 7% were larvae. Overall, 20% of 74 examined females and 9% of 203 examined nymphs (but none of 21 examined larvae) were infected with borreliae. The proportion of ticks with a high infection load (>100 spirochetes) was 4% in females and 2% in nymphal I. ricinus. During the year, the highest numbers and proportions of infected nymphal and female ticks were taken from humans in June. Detection of borreliae in the ticks feeding on humans might be helpful in the prophylaxis of Lyme borreliosis.     

Longitudinal surveillance of the tick Ixodes ricinus for borreliae

Host-seeking Ixodes ricinus (L.) (Acari: Ixodidae) were monitored for borreliae (Borrelia burgdorferi s.l.) using dark-field microscopy in South Moravia (Czech Republic) each May from 1991 to 2001 (150 nymphs, 100 females and 100 males each year). This survey revealed a mean annual percentage of infected ticks of 16.8% (range, 11.7-24.2) in nymphs, 24.9% (range, 16.5-33.6) in females and 26.1% (range, 17.1-37.3) in males. Annual incidence of Lyme borreliosis in humans of the area in the same period (range, 8.7-41.7 per 100,000) correlated significantly with the frequency (number of ticks per flag per hour) of nymphs infected with >50 borreliae or all nymphal ticks, but not with the frequency of females, infected females or the infection rate (% of ticks infected) of either nymphal or female ticks. A prediction of the annual incidence of Lyme borreliosis, based on the frequency of heavily infected or all nymphal I. ricinus ticks, is feasible. The infection rate in I. ricinus correlated significantly with the North Atlantic Oscillation winter index of the last year (in nymphs) or of the year before last (in adults).     

An electron microscopic study of Borrelia in the body of the female ixodid tick Ixodes persulcatus

Borrelia burgdorferi s. lato in naturally infected females of tick Ixodes persulcatus were examined by transmission electron microscopy. The Borreliae were found in midgut and ovary. Location and ultrastructure of bacteria indicate extracellular migration through the midgut epithelium as a preferential way. In gonad, the borreliae intracellular situate in ovarian epithelium and oocytes before and at the beginning of vitellogenesis. The demonstration of numerous spirochetes in the oocytes provides the support for transovarial transmission of the agent. Two morphological types of borreliae were observed.     

Amplification of tick-borne encephalitis virus infection during co-feeding of ticks

Abstract. Following engorgement of Rhipicephalus appendiculatus larvae on guinea-pigs infected with tick-borne encephalitis (TBE) virus, none of the engorged larvae or emergent nymphs contained detectable infectious virus. However, one of twelve pools, each containing three of the unfed nymphs, was positive when screened by polymerase chain reaction (PCR), indicating a low prevalence of TBE virus infection in the unfed nymphs. After engorgement of the nymphs on four uninfected guinea-pigs, 19/24 (79%) fed nymphs from one guinea-pig and 4/25 (16%) fed nymphs from a second guinea-pig were infected; all the ticks examined from the other two guinea-pigs were uninfected. The results suggest that TBE virus was transmitted from a low proportion of infected nymphs (infected as larvae) to uninfected nymphs as they fed together on an uninfected guinea-pig. Such amplification of the initial infection, at the population level, could play an important role in maintaining TBE virus infections in nature, particularly if there is a low level of vertical transmission from one tick generation to the next.     

Seasonal patterns of abundance and host relationships of the Australian paralysis tick,Ixodes holocyclus Neumann (Acarina: Ixodidae), in southeastern Queensland

Estimates of seasonal abundance of larvae, nymphs and females of the paralysis tick, Ixodes holocyclus, were obtained by collecting ticks that engorged on small mammals and birds trapped in two localities in southeastern Queensland: Brisbane (wet sclerophyll forest) and Tamborine Mountain (cleared rain forest). The long-tailed short-nosed bandicoot, Isoodon macrourus, was the most common mammal trapped but small numbers of other marsupials, rodents and ground frequenting birds were also captured. Small numbers of five other tick species were also collected. In both habitats there was clearly one dominant generation of I. holocyclus;7er year, although the presence of all stages at most times of the year indicated overlapping of smaller cohorts. Females were most abundant in spring and early summer, larvae in summer-autumn, and nymphs in autumn-winter. I. holocyclus was abundant on I. macrourus and rare on most other mammals and birds captured. At the peak of abundance of each instar, each bandicoot dropped from 500 to 2000 engorged larvae, 100 to 200 engorged nymphs, and four to six engorged females. Life tables were compiled for the tick in both habitats and these indicate that there was relatively high survival from engorged larva to engorged nymph and thence to engorged female and that most mortality occurred between detaching of the engorged female and the detaching of the engorged larva. The tick was more abundant on bandicoots from cleared rain forest and rain forest edge, than on those from sclerophyll woodland. The survival of engorged larvae and nymphs of I. holocyclus and the larval productivity of engorged females were examined in a warm moist climate where the tick was abundant (Tamborine Mountain) and in a hotter dryer climate where the tick was rare or absent (Amberley). In both localities engorged larvae and nymphs survived to the next instar in all seasons of the year. On most occasions engorged females produced eggs which hatched. Mature bandicoots from tick infested areas showed little or no resistance to infestation with larvae or nymphs of I. holocyclus, whereas other small mammals from the same area showed an appreciable degree of resistance to the immature stages of the tick. Feeding larvae and nymphs exposed to normal light-dark cycles in the laboratory detached during the afternoon and early evening. This behaviour and host resistance are discussed in relation to the daily activity cycles of host species, their habitat preferences, and their role as hosts for I. holocyclus.     

Experimental infection of the tick Amblyomma cajennense, Cayenne tick, with Rickettsia rickettsii, the agent of Rocky Mountain spotted fever

In the laboratory, Amblyomma cajennense (Acari: Ixodidae) (Fabricius) larvae, nymphs and adults were exposed to Rickettsia rickettsii by feeding on needle-inoculated animals, and thereafter reared on uninfected guinea pigs or rabbits. Regardless of the tick stage that acquired the infection, subsequent tick stages were shown to be infected (confirming transstadial and transovarial transmissions) and were able to transmit R. rickettsii to uninfected animals, as demonstrated by serological and molecular analyses. However, the larval, nymphal and adult stages of A. cajennense were shown to be partially refractory to R. rickettsii infection, as in all cases, only part of the ticks became infected by this agent, after being exposed to rickettsemic animals. In addition, less than 50% of the infected engorged females transmitted rickettsiae transovarially, and when they did so, only part of the offspring became infected, indicating that vertical transmission alone is not enough to maintain R. rickettsii in A. cajennense for multiple generations. Finally, the R. rickettsii-infected tick groups had lower reproductive performance than the uninfected control group. Our results indicate that A. cajennense have a low efficiency to maintain R. rickettsii for successive generations, as R. rickettsii-infection rates should decline drastically throughout the successive tick generations.     

The exploratory activity of the Borrelia-infected taiga tick Ixodes persulcatus

The infection with Borrelia burgdorferi s. l. increases the questing activity of adults and nymphs of the taiga tick Ixodes persulcatus. High temperature inhibits the moving and questing activities of the infected ticks in a greater extent than that in the non-infected ones. Than more borreliae per a specimen of tick are present, the more tick's activity is affected.     

Stage-specific population dynamics of cottony cushion scale,Icerya purchasi (Hemiptera: Monophlebidae), in citrus orchards in Jeju,Korea

The cottony cushion scale, Icerya purchasi Maskell (Hemiptera: Monophlebidae), is a polyphagous, cosmopolitan and destructive pest of citrus. This study was conducted to obtain the stage-specific phenology of I. purchasi for seasonal management strategies in the field. Movement of crawlers (hatched nymphs) in egg sacs of overwintered females started in late May, peaked in early to mid-June, and was completed by late June. Generally, the 1st generation occurred from late May until late September and the overlapping second generation occurred from early September. The 1st nymphs in the 1st generation peaked in mid June. The 2nd nymphs showed peak activity in late July. The 3rd nymphs showed peak population in early September. The 1st generation adults peaked in mid September. In the 2nd generation, the 1st nymphs peaked in early October, the 2nd nymphs showed peak activity in late October, and the 3rd nymphs reached a plateau after mid October. The 2nd generation adults occurred from late October. Consequently, two life cycles were competed in the Jeju area. The average fertilities of I. purchasi were 623 and 247 crawlers per female in overwintered and summer generations, respectively. An average of 20.7% of all citrus orchards was infested with I. purchasi, with a mean of 3.9% infested trees in Jeju. These results should be useful in establishing management strategies for I. purchasi in citrus orchards.     

Life tables and population parameters of the green leafhopper,Nephotettix cincticeps (Uhler), in the southwestern district of Japan with special reference to the first generation on foxtail grass

The three year (1988–1990) life tables of Nephotettix cincticeps were constructed, and the daily survival rate and longevity of female adults were estimated by Hokyo and Kiritani's (1967) method for the overwintering and the first-generations on the foxtail grass in Okayama, southwestern Japan. The life tables and the population parameter values estimated were compared with those in the other generations on rice. The FARMCOP suction sampler was employed to survey the population density. The durations of pre-ovarial maturation of female adults of the 1st generation on foxtail grass and rice seedling were similar. Longevity of adults of the overwintering and the first generations which emerged on the wild host was longer than that of the other generations (2nd and 3rd generations) on the rice plants. Fecundity of females decreased successively as the generation proceeded and it became lowest in the final 3rd generation. Only about 3.5 percent of first-instar nymphs of the 1st generation emerged as adults in the fallow field. The survival rate of nymphs on foxtail grass was always lower in comparison with that on rice plants. However, the survival rates of nymphs on foxtail grass and rice seedling were not significantly different from each other under laboratory conditions. In the fields, senescence of foxtail grass occurred in the midst of nymphal period of the 1st generation. The survival rate of nymphs on foxtail grass decreased with the increasing in the nymphal density. Abundance of spiders during the 1st generation was higher than that in the early stage of rice plants.     

Growth of Haemaphysalis (Kaiseriana) longicornis on cattle.

Increases were recorded of lengths of larvae, nymphs and females of the tick, Haemaphysalis (Kaiseriana) longicornis during feeding on cattle, so that categories could be established for estimating the ages of ticks in natural populations. Most larvae and nymphs engorged in 4 days and females in 5 or 6 days. Four age categories were defined for larvae and 3 for nymphs. The growth of females was variable but 4 stages of physiological development were defined based on changes in their lengths. The lengths were determined of each instar 24 h before detachment to enable estimation of the numbers maturing each day on cattle in the field.     

Entomopathogenic fungi associated with Ixodes ricinus ticks

The objective of this study was to demonstrate the occurrence of entomopathogenic fungi on Ixodes ricinus ticks in relation to the tick stage, engorgement and season. Ticks were collected from the vegetation, from small rodents and from deer. All entomopathogenic fungi found belonged to the Hyphomycetes. Paecilomyces farinosus and Verticillium lecanii were the predominant species. Other species, found only on engorged females were: Beauveria bassiana, B. brongniartii, P. fumosoroseus and V. aranearum. Eight out of 1833 ticks collected from the vegetation and three out of 269 engorged nymphs were infected with fungi. Thirty-three out of 149 engorged females were infected, whereas males and engorged larvae were not infected. Throughout the season, a significantly higher proportion of ticks collected in autumn were infected. Entomopathogenic fungi may have a significant impact on the size of the I. ricinus population, since females were the most frequently infected stage.     

The role of medium-sized mammals as reservoirs of Borrelia burgdorferi in southern New York

The ability of raccoons (Procyon lotor), striped skunks (Mephitis mephitis) and opossums (Didelphis virginiana) to serve as reservoirs of Borrelia burgdorferi, the spirochetal agent of Lyme disease, was compared with that of white-footed mice (Peromyscus leucopus). Twenty-eight (28) medium-sized mammals and 34 white-footed mice were captured in Westchester County, New York (USA) in summer 1986. Animals were caged over pans of water for 1 to 2 days to recover engorged tick larvae (Ixodes dammini) that detached from the hosts after feeding. With the exception of mice, numbers of engorged tick larvae recovered exceeded those counted during initial examinations of the hosts by 30% (opossums) to nearly 90% (raccoons). Newly-molted nymphal ticks derived from the engorged larvae were examined for the presence of spirochetes by darkfield microscopy. Percentage infection was 5% (n = 22) for ticks from skunks and 14% (n = 191) for ticks from raccoons. None of 24 nymphs from larvae that fed on opossums survived long enough for spirochete examination. By comparison, 40% (n = 72) of nymphs from larvae which fed on white-footed mice were infected. Of the individual hosts from which molted nymphs had fed as larvae, 67% of mice, 33% of skunks, and 55% of raccoons produced spirochete-positive ticks.     

Comparative activity of three repellents against the ticks Rhipicephalus sanguineus and Argas persicus

The residual repellent activity of N,N-diethylphenyl-acetamide (DEPA), N,N-diethyl-3-methyl benzamide (DEET) and dimethylphthalate (DMP) against the hard tick Rhipicephalus sanguineus (Latreille) and the soft tick Argas persicus (Oken) from topical application on rabbits and hens respectively in the laboratory have been studied. It was found that a 25% concentration of DEET gave strong repellency (greater than 90%) against larvae and nymphs of R. sanguineus for 15 and 7 days respectively and against A. persicus larvae and nymphs greater than 90% for 11 and 5 days respectively. Strong repellency (greater than 90%) against adults of the hard and soft ticks was observed for 9 and 7 days respectively with 40% DEET. DEPA at 25% concentration showed strong repellency (greater than 90%) against larvae and nymphs of R. sanguineus and A. persicus for 11 and 5 days respectively. Adults of hard and soft ticks were strongly repelled (greater than 90%) for 9 and 5 days respectively by 40% DEPA. DMP was found to be the least effective against all stages of both species of tick.     

Microbiome changes through ontogeny of a tick pathogen vector

Blacklegged ticks (Ixodes scapularis) are one of the most important pathogen vectors in the United States, responsible for transmitting Lyme disease and other tick‐borne diseases. The structure of a host's microbial community has the potential to affect the ecology and evolution of the host. We employed high‐throughput sequencing of the 16S rRNA gene V3‐V4 hypervariable regions in the first study to investigate the tick microbiome across all developmental stages (larvae, nymphs, adults). In addition to field‐collected life stages, newly hatched laboratory‐reared larvae were studied to determine the baseline microbial community structure and to assess transovarial transmission. We also targeted midguts and salivary glands due to their importance in pathogen maintenance and transmission. Over 100 000 sequences were produced per life stage replicate. Rickettsia was the most abundant bacterial genus across all sample types matching mostly the Ixodes rickettsial endosymbionts, and its proportion decreased as developmental stage progressed, with the exception of adult females that harboured a mean relative abundance of 97.9%. Laboratory‐reared larvae displayed the lowest bacterial diversity, containing almost exclusively Rickettsia. Many of the remaining bacteria included genera associated with soil, water and plants, suggesting environmental acquisition while off‐host. Female organs exhibited significantly different β‐diversity than the whole tick from which they were derived. Our results demonstrate clear differences in both α‐ and β‐diversity among tick developmental stages and between tick organs and the tick as a whole. Furthermore, field‐acquired bacteria appear to be very important to the overall internal bacterial community of this tick species, with influence from the host bloodmeal appearing limited.     

Effect of forest clearing on the abundance of Ixodes ricinus ticks and the prevalence of Borrelia burgdorferi s.l

Questing Ixodes ricinus L. (Acari: Ixodidae) ticks were collected on a forest trail that had been completely cleared of shrubs and ground vegetation in winter 2002 and on a nearby control uncleared forest transect in South Moravia (Czech Republic). Samples were collected each May in 2003, 2004 and 2005. Nymphal ticks were 3.4 times, 1.9 times and 1.2 times less frequent on cleared forest than on uncleared forest trails in the three respective years, whereas adult tick abundance was 27.2 times, 4.0 times and 2.2 times lower, respectively. The ticks were examined for borreliae by dark-field microscopy: prevalence of nymphal ticks infected with Borrelia burgdorferi sensu lato (12.6% to 20.0%) did not differ significantly between the cleared and uncleared trail during the 3 years. In conclusion, the habitat modification appeared to result in a decreased abundance of I. ricinus as well as a reduced frequency of infected ticks (and thus indirectly a lower potential risk of Lyme borreliosis), which lasted, however, for only 2 years. Eight cultures of borreliae isolated from the ticks were all identified as the 'ornithophilic' genomic species Borrelia garinii, possibly indicating a greater role of forest birds than that of forest rodents as the hosts of immature I. ricinus in the tick (and borrelial) colonization of the cleared part of the forest.     

The efficiency of patch sampling for determination of relative tick burdens in comparison with total tick counts

Quanttative data on host tick burdens ar fundamental for the initiation of control strategies and effective management of wildlife populations, but the methods of live sampling employed for domestic animals are unsuitable for sampling wild animals. Despite advances in the use of destructive methods (the scrub and digestion techniques) to obtain measures of the total tick burden on wildlife, these methods are too involved for many field workers, who often need only measures of relative tick burden. Recently, patch sampling methods have been introduced whereby only certain predilection sites are sampled, the presumption being that the number of ticks collected gives an indiccation of the relative degree of infestation. We examined the validity of patch sampling as a measure of relative tick burden by comparing adult ticks collected from the ears, head, neck, foreleg and perianal region of impala (aepyceros melampus) with total tick burdens of the same animals derived from the digestion technique. Adult ticks from patch sampling were positively and significantly correlated with total adults and total ticks (larvae, nymphs, and adults) on impala, with ticks patch sampled from the neck showing the highest correlation with the total tick burden. Comparison of relative tick loads from patch sampling with absolute tick loads from digestion for three classes of impala (females, bachelor males and territorial males) gave qualitatively similar results. We conclude that, when measures of relative tick load are sufficient and destructive sampling is not feasible, patch sampling can provide reliable information on relative tick burdens that are positively correlated with the total tick burden.     

Prevalence of Theileria in the tick Hyalomma detritum detritum in the Doukkala region,Morocco

Abstract. The overall prevalence of Theileria species, mainly, if not exclusively, T.annulata, in 901 Hyalomma detritum detritum collected from cattle in the Doukkala region of Morocco over a period of 2 years was 21.5%. The quantity of infection (number of sporoblasts per infected tick) followed the negative binomial distribution with between one and 250 sporoblasts per infected tick. Infected ticks were found in eight of fourteen areas examined whilst T.annulata was present in all fourteen. There were significant differences in both the prevalence and the quantity of infection between ticks collected from different farms, and between nymphs collected in the autumn from these farms, and moulted in the laboratory, and adults collected in the following summer. The prevalence, but not the quantity, of infection was higher in female than in male ticks. No correlations were established between infection of engorged nymphs and the breed, sex and Theileria piroplasm parasitaemia of the host animal. However, calves infected a greater proportion of nymphs than adult cattle and the heavier the infestation of nymphs on an animal, up to a plateau, the higher the prevalence of infection in those nymphs. There were no differences in infection between ticks moulted at 24^oC and 37^oC, after the engorged nymphs had been stored at 18^oC to simulate over-wintering.     

Tick burden on European roe deer (<Emphasis Type="Italic">Capreolus capreolus</Emphasis>)

In our study we assessed the tick burden on roe deer (Capreolus capreolus L.) in relation to age, physical condition, sex, deer density and season. The main objective was to find predictive parameters for tick burden. In September 2007, May, July, and September 2008, and in May and July 2009 we collected ticks on 142 culled roe deer from nine forest departments in Southern Hesse, Germany. To correlate tick burden and deer density we estimated deer density using line transect sampling that accounts for different detectability in March 2008 and 2009, respectively. We collected more than 8,600 ticks from roe deer heads and necks, 92.6% of which were Ixodes spp., 7.4% Dermacentor spp. Among Ixodes, 3.3% were larvae, 50.5% nymphs, 34.8% females and 11.4% males, with significant seasonal deviation. Total tick infestation was high, with considerable individual variation (from 0 to 270 ticks/deer). Adult tick burden was positively correlated with roe deer body indices (body mass, age, hind foot length). Significantly more nymphs were found on deer from forest departments with high roe deer density indices, indicating a positive correlation with deer abundance. Overall, tick burden was highly variable. Seasonality and large scale spatial characteristics appeared to be the most important factors affecting tick burden on roe deer.     

Instar development and disengagement rate of engorged female winter ticks,Dermacentor albipictus (Acari: Ixodidae), following single- and trickle-exposure of moose (Alces alces)

Seven hand-reared moose (Alces alces) calves and one yearling were infested with 30 000 larvae each of the winter tick,Dermacentor albipictus, either by single- or trickle-exposure (1000/day). They were examined weekly for instar changes from September/October until late May. By 2 and 3 weeks post-exposure, most larvae on single- and trickle-infested moose, respectively, had fed and molted to nymphs. Thereafter, tick development was similar between both infestation techniques. Nymphs dominated the tick population from October to mid-February, and adults from mid-February to May. The peak of host disengagement by engorged females was late March in both years. Weights of engorged females from calf moose declined over time during the disengagement period.     

Histological examination ofBorrelia burgdorferi infections in unfedIxodes ricinus nymphs

Borrelia burgdorferi, the agent of Lyme borreliosis, is vectored in Europe byIxodes ricinus. In unfed ticks, the spirochaete resides primarily in the midgut, but a low percentage (5.5%; 4/73) of naturally infected ticks may present a systemic infection involving organs such as the salivary glands and central ganglion (Burgdorferet al., 1983). In this study, we examined 79 unfed nymphs collected in two sites in Switzerland (Neuchatel and Aarberg), and 35 unfed nymphs which were fed as uninfected larvae on 3 infectedApodemus sylvaticus mice. Dieterle silver staining was used to visualize the spirochaetes in the ticks. Of the unfed field-collected nymphs, 21/79 (27%) were infected of which 2/21 (10%) had systemic infections. Taking account of the site of collection, we observed that 0/12 ticks from Neuchatel were systemically infected whereas 2/9 (22%) from Aarberg had a disseminated infection. Out of the 35 unfed nymphs examined after an infectious blood meal on rodents, 14 (40%) were infected and 2 (14%) had a disseminated infection. A total of 4/35 (11%) unfed infected nymphs presented a systemic infection which represents a higher percentage than previously described. The presence of spirochaetes in salivary glands of systemically infected ticks before the initiation of feeding may reduce the time delay generally recorded for the tick-borne transmission ofB. burgdorferi.