Seasonal dynamics and hosts of Amblyomma triste (Acari: Ixodidae) in Argentina

The seasonal dynamics and host usage of Amblyomma triste in Argentina were analyzed. Adults of A. triste were present from early winter to mid-summer, with the peak of abundance from late winter to mid-spring (August to October). Larvae and nymphs were found from December to June, with the peak of abundance in summer. There were no differences among the biological parameters (pre-moult period of larvae and nymphs, pre-oviposition period of females, and minimum incubation period of eggs) of engorged ticks exposed to different photoperiod regimens at the laboratory, but the periods for each biological parameter obtained from ticks exposed in the field were significantly longer than those from the laboratory. Field results fit better with the data of seasonal distribution of each stage. Morphogenetic diapause was not detected, but complementary studies should test the presence of behavioral diapause. Rodents of the subfamily Sigmodontinae (Akodon azarae, Oligoryzomys flavescens, Oligoryzomys nigripes, Oxymycterus rufus and Scapteromys aquaticus) are the principal hosts for immature stages of A. triste, the caviid Cavia aperea could be another potential host for these stages, and birds are exceptional hosts for larvae and nymphs. Regarding hosts of adults in Argentina, domestic and wild large-sized mammals belonging to different orders (cattle, dog, horse, Blastocerus dichotomus and Hydrochoerus hydrochaeris) serve as hosts for adults of this tick species. In conclusion, A. triste has a life cycle of 1 year with adults feeding on large endemic and introduced mammals and immature stages using sigmodontine and caviid rodents as hosts.     

Larval Gulf Coast ticks (Amblyomma maculatum) [Acari: Ixodidae] as host for Hepatozoon americanum [Apicomplexa: Adeleorina].

Laboratory-reared larval Gulf Coast ticks (GCTs) (Amblyomma maculatum) were exposed experimentally and found to acquire Hepatozoon americanum infection while feeding on parasitemic dogs. These ticks supported gamogonic and sporogonic development of the apicomplexan, and oocysts from newly molted nymphs were infectious for a dog. Other nymphs from this cohort that were allowed to feed on a blood-parasite naive sheep molted normally; the resulting adult ticks contained oocysts that were infectious for another dog. Merogonic development of H. americanum in the dogs and the resulting lesions/disease appeared similar, irrespective of whether infectious oocysts were derived from nymphal or adult ticks that acquired infection as larvae. In the system previously known, nymphal ticks acquire infection and adults harbor infective oocysts, which vertebrate hosts ingest. Given that larval A. maculatum can acquire infection and nymphs can harbor viable oocysts as demonstrated by this study, the potential variety of vertebrate hosts that can alternate with GCTs in maintaining an endemic cycle is considerably expanded.     

Hyalomma anatolicum anatolicum (Koch, 1844) as a possible vector for transmission of Trypanosoma theileri,Laveran, 1902 in cattle

Amastigotes, sphaeromastigotes, epimastigotes and trypomastigotes of Trypanosoma theileri, Laveran, 1902 have been observed in the nymphs and adults of Hyalomma anatolicum anatolicum (Koch, 1844). These ticks were obtained from engorged larvae and nymphs collected from a cross-bred bull with detectable parasitaemia. Infection with T. theileri was established inoculation of Trypanosomatid flagellates collected from ticks and by feeding of the infected ticks on the ears of these calves.     

Ecological aspects of four species of ticks found on cattle in Salta, northwest Argentina

The seasonal activity of Amblyomma cajennense, Amblyomma neumanni, Amblyomma parvum and Boophilus microplus on cattle was studied in a ranch located in the northern part of the phytogeographical district of the Chaco Serrano where the habitat had suffered minimal human disturbance. The female ticks on one side of 10-25 cows were counted at intervals of 20-50 days from 29 October 1985 to 5 November 1986. The percentage of cattle parasitized with larvae and nymphs of Amblyomma was recorded, along with the proportion corresponding to the different species. Collection of ticks from the ground cover vegetation in areas with and without forest was performed using the dragging technique. B. microplus was found all year round; peaks of abundance were observed in May (55 females per cow/2) and November 1986 (71 females per cow/2). The peak in November was unexpected and possibly due to a higher stocking rate around watering places and poor nutrition at the end of the dry season. A. cajennense appears to produce a generation a year. Larvae were more abundant from June to August, nymphs from August to November and females from November (34 per cow/2) to January (15 per cow/2). A. neumanni showed a cycle corresponding to a generation every 3 years regulated via diapause that involves, at least, all the summer months. The peak of abundance for larvae, nymphs and females occurred in late autumn-early winter. The highest number of females (44 per cow/2) was recorded in June.(ABSTRACT TRUNCATED AT 250 WORDS)     

Borrelia burgdorferi sensu lato and Rickettsia spp. infections in hard ticks (Ixodes ricinus) in the region of Hanover (Germany)

In a total of 605 Ixodes (I.) ricinus ticks collected in the spring-months March, April and May 2005, quantitative real time PCR (qPCR) revealed 26.6% Borrelia (B.) burgdorferi sensu lato (sl)-positive ticks, i. e. divided by sex and stage into 31.9% positive adults (34.8% females and 29.0% males) and 18.5% positive nymphs. Mono-infections with genospecies from the B. burgdorferi sl-complex were found in over two thirds of the positive individuals, whereas almost one third showed double- or even triple-infections. Genospecies-specific conventional PCR determined B. afzelii as the most frequent genospecies followed by B. garinii, B. spielmanii, B. valaisiana and B. burgdorferi sensu stricto (ss). Rickettsia spp. were found in 34.2% of the collected ticks, divided into 37.6% adults (42.5% females and 32.8% males) and 29.0% nymphs. Co-infections of Rickettsia-positive ticks with B. burgdorferi sl spirochaetes were present in 10.1% of the ticks. Thereby, adult ticks exhibited a co-infection rate of 13.4% (15.5% females and 11.3% males) and nymphs of 5.0%. Independently of the above mentioned study, 3939 Ixodes ticks, sent in between 2006 and 2010 for B. burgdorferi sl-diagnostic, were examined by qPCR exclusively for B. burgdorferi sl. The resulting B. burgdorferi sl prevalence was 23.1% and 24.4% in 2006 and 2007, respectively, followed by a continuous decrease to 12.8% in 2010. To analyse whether this observed decrease in infection frequency is due to sampling bias, in a current study randomly sampled ticks collected from defined sites equally distributed over the city of Hanover are investigated in a statistically relevant sample size.     

Laboratory and field observations on anti-tick properties of the plant Gynandropsis gynandra (L.) Brig.

A shrubby plant, abundant in east Kenya, Gynandropsis gynandra (L.) Brig., was shown to exhibit repellent and acaricidal properties to larvae, nymphs and adult Rhipicephalus appendiculatus and Amblyomma variegatum ticks. All stages of ticks avoided the leaves of the plant and a high percentage of the ticks which were continuously exposed to its leaves died; surviving ticks were weak and inactive but regained activity when exposed to fresh air. The effectiveness of the plant's leaves as a repellent and acaricide was most pronounced on nymphs and least pronounced on adults. Field investigations indicated that ticks were not found up to 2-5 m from the plant in areas where the plant was predominant. The potential of using the plant for tick control within an integrated tick management system in the resource-poor farming context in Africa was highlighted.     

Experiments on transmission of an unidentified Theileria sp. to small ruminants with Haemaphysalis qinghaiensis and Hyalomma anatolicum anatolicum

Experiments on the transmission of an unidentified Theileria sp. infective for small ruminants by Haemaphysalis qinghaiensis and Hyalomma anatolicum anatolicum were carried out. Three Theileria-free batches of adult, larvae, and nymphs of laboratory reared H. qinghaiensis and Hy. a. anatolicum ticks were infected by feeding them on sheep infected with Theileria sp. The Theileria sp. was originally isolated from adult ticks of H. qinghaiensis, by inoculation of blood stabilates or tick transmission. H. qinghaiensis has been shown to be capable of transmitting the Theileria sp. infective for small ruminants transstadially to sheep and goats. The nymphs developed from the larvae engorged on the sheep infected with the parasite transmitted the pathogen to splenectomized sheep with prepatent periods of 30, 31 days, respectively; but the subsequent adult ticks of H. qinghaiensis derived from the nymphs did not transmit the pathogen to sheep. However, adults developed from the nymphs engorged on the sheep infected with the parasite transmitted the pathogen to sheep with prepatent periods of 24-27 days. The larvae, nymphs and adult ticks derived from female H. qinghaiensis ticks engorged on infected sheep were not able to transmit the parasite transovarially. The same experiments were done with Hy. a. anatolicum, but examination for presence of piroplasma of Theileria sp. from all animals were negative, demonstrating that Hy. a. anatolicum could not transmit the organism to sheep or goats.     

Rhipicephalus sanguineus (Ixodida, Ixodidae) as intermediate host of a canine neglected filarial species with dermal microfilariae

The life cycles of filarioids of dogs presenting dermal microfilariae have been little studied. Following the recent retrieval of dermal microfilariae identified as Cercopithifilaria sp. in a dog from Sicily (Italy), this study was designed to assess the role of the brown dog tick Rhipicephalus sanguineus as an intermediate host of this filarial species. An experimental tick infestation was performed on an infected dog using 300 nymphs of R. sanguineus. Engorged nymphs were collected and examined by both microscopic dissection and molecular analysis at five time points (i.e., the same day of tick detachment and 10, 20, 30 and 50 days post-detachment) to detect the presence and developmental stage of filariae in the ticks. A total of 270 engorged nymphs were collected from the dog and developing filarioid larvae detected in 10 (5%) out of 200 ticks dissected. Infective third-stage larvae were observed in 4 (2%) of the all dissected ticks, 30 days post-detachment. Twelve (6.6%) out of 181 samples molecularly tested were positive for Cercopithifilaria sp. This study demonstrates that nymphs of R. sanguineus feeding on a dog naturally infected by Cercopithifilaria sp. can ingest microfilariae, which develop up to the third infective stage thus suggesting that this tick species might act as an intermediate host of this little known canine filarioid.     

Saturation deficit and deer density affect questing activity and local abundance of Ixodes ricinus (Acari, Ixodidae) in Italy

The wood tick Ixodes ricinus, one of the most common arthropod-borne disease vectors, is of increasing relevance for human and animal health in Europe. The aim of this study was to determine the relative contribution of several abiotic and biotic factors potentially affecting questing activity and local abundance of I. ricinus in Italy, considering the scale at which these factors interact with the host-seeking ticks. Within EDEN, a large-scale EU collaborative project on eco-epidemiology of vector-borne diseases, we collected questing ticks for three consecutive years using a standard protocol at eleven sites in the Italian Alps and Apennines. A total of 25 447 I. ricinus were collected. All sites showed the same annual pattern of tick activity (bimodal for nymphs and unimodal for larvae and adults), although the abundance of nymphs was statistically different between sites and years. A Generalized Linear Mixed Model and a Linear Mixed Model fitted to data for nymphs, showed that while the principal variables affecting the local abundance of questing ticks were saturation deficit (an index combining temperature and relative humidity) and red deer density, the most important variable affecting questing nymph activity was saturation deficit. As for the timing of seasonal emergence, we confirmed that the threshold temperature at this latitude for larvae is 10°C (mean maximum) while that for nymphs is 8°C.     

The natural hosts of larvae and nymphs of Amblyomma tigrinum Koch, 1844 (Acari: Ixodidae)

The hosts of larvae and nymphs of Amblyomma tigrinum, a tick whose adults feed on wild and domestic Canidae in South America, are uncertain. A 17 months survey was carried out trapping wild vertebrates in north-western Córdoba, Argentina, to evaluate their parasitism with A. tigrinum subadults. Larvae and nymphs of this tick species were identified conventionally and by comparison of 16S rDNA sequences with GenBank deposited sequences. A total of 207 small and medium-sized rodents and 182 birds were captured and examined for ticks. Most ticks on birds were from ground forest feeding birds (BB) with a minimal contribution of birds feeding in open pastures. All ticks from rodents were obtained from representatives of the families Cricetidae (SR) and Caviidae (MR). Percent of larvae infestation was higher (P<0.01, Chi-square distribution) in BB (55.2%) and SR (46.4%) than in MR (17.4%) and the same trend was found for number of larvae on these hosts (test of Kruskal-Wallis). Caviidae (only representative Galea musteloides) rodents were extremely prone to be infested with nymphs of A. tigrinum (94.2%) followed by BB (50.6%) and SR (3.6%) (P<0.01) and the same tendency was found for number of nymphs (P<0.01). The index of aggregation for nymphs on MR was the lowest (0.409) followed by nymphs on BB (0.706) which may be a consequence of higher and homogenous exposure of G. musteloides to host-seeking nymphs. Several BB are food source for both larvae and nymphs of A. tigrinum while for rodents larvae were common only on SR (mainly on the Sigmodontinae Akodon dolores and Graomys sp.) and nymphs feed almost exclusively on MR. Therefore, both birds and rodents are of importance for the survival strategy of A. tigrinum subadults. The plasticity of A. tigrinum to colonize areas with different climates plus the capacity of their subadults to feed on hosts widely distributed indicates that this tick has the potential to become a widespread parasite but this does not seem to be the actual situation. Several proposals are presented to further understand its ecology.     

Percutaneous infection of nymphal Dermacentor andersoni with Anaplasma marginale

Newly replete nymphal Dermacentor andersoni (principals) were percutaneously exposed to Anaplasma marginale by injection of either intact or lysed infected bovine erythrocytes. Control nymphs were fed on calves with anaplasmosis. The subsequently molted adults were examined for infection by light microscopy, and companion ticks were tested for infectivity by allowing them to feed on susceptible calves. When they fed as adults, both control ticks and percutaneously inoculated principals transmitted A marginale to susceptible calves. Prepatent periods in calves varied according to the method by which nymphs were infected. Colonies of A marginale were found in all ticks that acquired infection by feeding, but colonies were not observed in any ticks exposed percutaneously. The possible developmental cycle of A marginale in artificially infected ticks is discussed.     

INFECTIVITY FOR CATTLE OF ANAPLASMA MARGINALE EXTRACTED FROM BOOPHILUS MICROPLUS TICKS EXPOSED TO CERTAIN TEMPERATURES

SUMMARY: Boophilus microplus ticks collected from calves with patent Anaplasma marginale infections were incubated at either 4 to 5°C, 14°C, 22°C, 27°C or 37°C for up to 14 days. Extracts prepared either from larvae, nymphs, immature females, adult males or mixtures of both sexes were infective for 14 of the 16 splenectomised calves inoculated. Extracts either from nymphs or from adult ticks deriving from nymphs moulted in vitro were infective for 11 of 12 nonsplenectomised calves. Possible application of the findings to producing a vaccine strain of A. marginale is discussed.     

Transmission of Ehrlichia canis to dogs by ticks (Rhipicephalus sanguineus).

Two strains of Rhipicephalus sanguineus acquired Ehrlichia canis by feeding as either larvae or nymphs on acutely infected dogs and, in subsequent instars, transmitted the agent to normal dogs. Three strains of R sanguineus transmitted E canis as adults after their larval and nymphal stages fed on infected dogs. More than 400 adult female ticks were fed on infected dogs as larvae or nymphs or both, but none transmitted E canis transovarially.     

Feeding predilection of Gulf Coast tick, Amblyomma maculatum Koch, nymphs on cattle

Gulf Coast tick nymphs successfully attached and fed on cattle after being freely released. Six Hereford heifers were each infested with approximately 2000 Gulf Coast tick nymphs, three with a strain originating from Refugio Co., TX, and three with ticks from Osage Co., KS by free release on the head and legs to simulate field acquisition of questing nymphs. Two re-infestations were conducted, the first at 7 days and the second at 28 days. Nymph dispersal was estimated by daily inspection of 22 body areas and removal of engorging ticks from the third to the fifth days post-infestation. Total recovery of engorging Texas nymphs was 3.0, 10.2, and 0% and Kansas nymphs was 21.5, 3.3, and 0% for infestations one, two and three, respectively. Immunological resistance to tick infestation expressed as cellular hypersensitivity was evident against Kansas nymphs in the second infestation and against both tick strains in the third infestation. Ticks removed from the withers, midline, and tail-head areas accounted for 68% of the total nymphs recovered in the first two infestations. Within these areas, nymphs were observed to aggregate in small spots where the hair was less dense or naturally parted and the remainder were found scattered in dense hair.     

Transstadial and attempted transovarial transmission of Anaplasma marginale by Dermacentor variabilis

Transstadial and transovarial transmission of Anaplasma marginale by Dermacentor variabilis were attempted with with ticks exposed to the organism once by feeding as larvae or nymphs, and twice by feeding as larvae and nymphs. Typical colonies of A marginale were in gut tissues of adults that were infected as larvae, larvae and nymphs, and as nymphs; repeated exposure of ticks did not appear to result in an increase in the number of colonies in the gut of subsequently molted adults nor did it affect severity of the clinical disease that developed in cattle they fed on. In contrast, colonies of A marginale were not found in the midgut epithelium of unfed nymphs exposed as larvae, even though companion nymphs transmitted the parasite, causing severe clinical anaplasmosis in susceptible calves. The organism was not transmitted transovarially by F1 larvae or nymphs from the groups exposed as parent larvae, nymphs, larvae and nymphs, and as adults. Some of the calves fed on by F1 progeny had a few erythrocytic marginale bodies that looked suspiciously like A marginale, as well as postchallenge exposure prepatent periods that were longer than other calves in the transovarial transmission study. Sera from these calves were tested for antibody to A marginale, using a highly sensitive immunoblot technique. Antibodies were not detected in any of the sera.     

Studies on the critical water mass, rehydration capability and potential, acute chill tolerance and supercooling point of Argas (Persicargas) walkerae (Acari: Argasidae)

The critical water mass, defined as the water mass remaining in a dehydrated tick in the non-ambulatory state, differed only slightly between light and heavy mass groups of Argas walkerae and averaged 23.6% and 23.2%, respectively, in males and 28.4% and 28.0%, respectively, in females. All ticks survived dehydration to 50%, 75% or 100% of their critical water mass, and 95% of them rehydrated during their subsequent incubation at 95% relative humidity (RH) and 28 degrees C for 14 days and regained their ambulatory status. Unfed adults were able to balance water loss frequently over a period of several months. When ticks were repeatedly dehydrated at 0% RH for 14 days, females and males suffered 50% mortality after 16 and 19 cycles of de- and rehydration, respectively, over a period of 278 days and 337 days, respectively. Water itself was not attractive to either dehydrated or non-dehydrated ticks and drinking was not observed. After submergence in water for 3 days, most of the dehydrated adult ticks gained mass. Judged by 50% mortality, larvae tolerated short-term extreme chilling to -24 degrees C, nymphs 1 to -22 degrees C, nymphs II to -20 degrees C, females and males to -19 degrees C. None survived tissue freezing. At a chilling rate of 0.3 degrees C/min, mean supercooling points (SCP) ranged from -25.9 degrees C in eggs to -16.5 degrees C in unfed females. The SCP of all other stages was significantly higher than that of eggs. Mean SCPs of unfed adult ticks dehydrated to 50% or 75% of their critical water mass were significantly lower than that of fully hydrated ticks. The SCPs of ticks acclimated by several weeks exposure to 0 degrees C or to 38 degrees C were significantly lower than those of adult ticks kept constantly at 28 degrees C.     

The effect of the bont tick (Amblyomma hebraeum) on the weight gain of Africander steers

The effects of the bont tick Amblyomma hebraeum on the productivity of cattle need to be quantified in order to design economically optimal control programs. Liveweight gains (LWGs) of three groups of Africander steers, maintained in the same pasture and exposed to zero, medium or high numbers of larvae, nymphs and adults of the tick, were measured. Larvae and nymphs had no significant effect on LWG but adults had a large, statistically significant effect (P less than 0.05). The tick-free group had an average LWG of 20 kg more than the heavily infested group after 3.5 months. No mortality was recorded from ticks or tick-borne diseases during the experiment. There was no relationship between the number of engorging adult female ticks counted and loss of LWG of individual animals. The latter suggests that exposure to ticks as well as engorgement by female ticks causes losses. The loss per adult female that completed engorgement was estimated, by relating the LWGs of individual animals to their tick infestations, to be 4 +/- 2 g per adult female tick. An alternative estimate, made by comparing the average LWG and tick infestations of each treatment group, was equal to 10 +/- 4 g. This latter estimate includes the effect of challenge (and rejection) as well as tick feeding and so was accepted as giving the best estimate. Screw-worm fly (Chrysomya bezziana) struck an average of 7.5% of the cattle infested with ticks in any week but the effects on the productivity of the cattle were minimized by immediate treatment. There was a significant, positive correlation between the incidence of screw-worm fly strike and the numbers of adult ticks counted on the experimental animals. The results provide data for calculating losses caused by A. hebraeum in different parts of its geographical range.     

The effects of bovine tick resistance on the susceptibility of Hyalomma anatolicum anatolicum to infection with Theileria annulata (Ankara)

Two Bos taurus calves were made resistant to tick infestation by exposing them to approximately 500 rabbit-reared nymphs of Hyalomma anatolicum anatolicum twice at a 2-week interval. These two calves, together with a tick-susceptible control calf, were inoculated with a stabilate of Theileria annulata (Ankara). Patent infection resulted in all three calves. Seven-hundred and fifty gerbil-reared nymphs were then applied on each of these calves as well as another tick-susceptible calf that was Theileria free. This infestation was carried out on Day 8 post-inoculation. Ticks that dropped on Day 13 post-inoculation were examined to note the development of T. annulata in them and the histological changes that occurred in the gut and salivary glands. During the second phase of feeding, the gut epithelia of the ticks from the tick-resistant calves were less active. There were no notable differences in the characteristics of the developmental stages of T. annulata between the ticks from the tick-resistant calves and those from the susceptible calf. However, ticks from one calf that acquired a higher level of tick resistance were significantly less susceptible to infection by T. annulata. Bovine tick resistance therefore compromises the vector capacity of H. a. anatolicum and this may be of epidemiological significance in the endemic areas of tropical theileriosis.     

Infectivity rate and transmission potential of Hyalomma anatolicum anatolicum ticks for Babesia equi infection

The infectivity rate of Babesia equi in the salivary glands of Hyalomma anatolicum anatolicum was assessed. The hungry nymphs were fed on a donkey experimentally infected with B. equi. The engorged dropped-off nymphs were collected at different levels of parasitaemia and kept in BOD incubator. After ecdysis, the hungry adults were prefed on rabbits for different time intervals, thereafter the salivary glands were dissected out and acini were examined after methyl green pyronin (MGP) staining. A total of 134 male and 139 female ticks were dissected out. Average infected acini per tick were found to be significantly higher (p<0.05) in male as compared to the female ticks. Further, maximum infected acini in both male and female ticks were found at 24h of prefeeding on rabbits and overall infected acini per tick increased with rise in parasitaemia. The release of infected ticks on susceptible donkeys resulted in development of clinical babesiosis.     

Resistance of restrained Bos taurus dairy bull calves to the bont tick Amblyomma hebraeum (Acarina: Ixodidae)

Three infestations of restrained Jersey and Friesland bull calves with adult Amblyomma hebraeum did not affect the rate of engorgement of female ticks and resulted in an insignificant decline in the mean engorgement weight of female ticks recovered from the Friesland group. Repeated infestations significantly affected the egg-laying capacity of engorged female ticks recovered from both groups of hosts. The Friesland calves manifested an enhanced resistance to the larvae and nymphs of Amblyomma hebraeum resulting in prolonged attachment, and a significant decline in engorgement rate of these instars compared with the Jersey group. Skin reaction tests using crude tick extracts yielded antigen-specific hypersensitivity reactions which were not related to resistance to A. hebraeum. Histological changes at the attachment site associated with tick rejection are discussed in relation to host resistance mechanisms.