Laboratory assessment of acaricidal activity of Cymbopogon winterianus,Vitex negundo and Withania somnifera extracts against deltamethrin resistant Hyalomma anatolicum

Larval packet test was used for detection of resistance levels against cypermethrin and deltamethrin, the most commonly used synthetic pyrethroids, in the multi-host tick Hyalomma anatolicum collected from district Moga, Punjab (India). Results indicated the presence of level I resistance against deltamethrin (RF = 2.81), whereas the tick isolate was susceptible to cypermethrin (RF = 0.2). The aqueous and ethanolic extracts of leaves of Cymbopogon winterianus, Vitex negundo and Withania somnifera along with roots of Vitex negundo were assessed for their acaricidal activity against the larvae of deltamethrin resistant H. anatolicum. The efficacy was assessed by measuring per cent larval mortality and determination of LC₅₀ values. The various ethanolic extracts produced a concentration dependent increase in larval tick mortality, whereas the aqueous extracts exhibited a much lower mortality. The highest mortality (93.7 ± 0.66 %) was observed at the 5.0 % concentration of ethanolic extract of leaves of C. winterianus and the lowest LC₅₀ value (0.011 %) was recorded for ethanolic extracts of leaves of V. negundo. The results indicated that these plant extracts have potential to be developed as herbal acaricides.     

Tick survey and detection of Crimean-Congo hemorrhagic fever virus in tick species from a non-endemic area, South Marmara region, Turkey

Crimean-Congo hemorrhagic fever (CCHF) is an increasing health concern in Turkey since 2002. There were also some recent human cases from the South Marmara region of Turkey; thus, a tick survey was performed, and possible vector tick species for the CCHF virus were determined in the region. A total of 740 adult ticks were collected from infested livestock from five locations: Çanakkale-Biga, Bursa-Orhaneli, Bursa-Keles, Bal?kesir and Bilecik. Total of 11 tick species from the genera Hyalomma, Rhipicephalus, Dermacentor, Ixodes and Haemaphysalis were identified. Rhipicephalus ticks were dominant in the region; the most frequently observed tick species was R. turanicus, (53.1 %), and only 15.4 % of the identified ticks were H. marginatum. The occurrence of H. rufipes infestation in the region fort he first time. A total of 73 pools of adult ticks were tested with both an antigen-detecting ELISA and RT real-time PCR (RT rt PCR). The presence of the CCHF virus was demonstrated in 9 (12.3 %) of the tested tick pools. Although seven of the tick pools were positive for the CCHF virus with both of the methods, one pool was positive only with RT rt PCR and the other pool was only positive with the ELISA. Positive results were obtained from ticks collected from cattle, sheep and goats from two locations, Bursa-Orhaneli and Bilecik. The CCHF virus was detected in R. turanicus (n = 3), R. bursa (n = 2), H. marginatum (n = 2) and D. marginatus (n = 2) ticks. The results of this study confirm the presence of the CCHF virus and present preliminary data on the vector tick species in the southern Marmara region of Turkey.     

In vitro efficacy of <Emphasis Type="Italic">Metarhizium anisopliae</Emphasis> sensu lato against unfed <Emphasis Type="Italic">Amblyomma parvum</Emphasis> (Acari: Ixodidae)

Amblyomma parvum Aragão (Acari: Ixodidae) is a tick species found with wide distribution in the Neotropical region. Even though it is a wildlife-related tick, it is also a frequent parasite of domestic animals, is aggressive to human beings and may harbor pathogenic microorganisms. Therefore, it is a target species for control on domestic animals, particularly those at the rural–wildlife interface. Herein, the efficacy of two isolates (E9 and IBCB 425) of an entomopathogenic fungus, Metarhizium anisopliae sensu lato, already evaluated for ticks that parasitize domestic animals, was tested against unfed A. parvum adults. Both isolates displayed high acaricidal efficacy after immersion in fungal conidial suspensions for 5 min. Isolate E9 killed all ticks by the 7th day post-treatment, and isolate IBCB 425 did so by the 11th day. Tick mortality of 80 and 90% was achieved as early as the 3rd and 4th days, respectively, with both treatments. Thus, if a commercial M. anisopliae s.l. acaricide against domestic animal ticks is developed, it would also be effective against A. parvum.     

Detecting Resistance to Organophosphates and Carbamates in the Cattle Tick Boophilus Microplus, with a Propoxur-based Biochemical Test

Rapid and sensitive detection of resistance to insecticides in arthropods is needed. In the cattle tick, Boophilus microplus, resistance to a variety of acaricides is widespread. The most commonly used assay for resistance, the larval packet test, takes at least two, but generally six weeks for a one-host tick like B. microplus to complete and may take up to three months to complete for three-host ticks. Here we describe a test for resistance to organophosphate acaricides that can be used on larvae and adult ticks which takes less than 24 hours. The test measures the difference in acetylcholinesterase (AChE) activity in homogenates of ticks in the presence and absence of propoxur, a carbamate acaricide. We found clear discrimination of organophosphate-susceptible and organophosphate-resistant adults with 100 M propoxur. AChE from susceptible ticks had almost no activity at this concentration of propoxur whereas AChE from resistant ticks had 67% of its potential activity. AChE from heterozygote ticks could also be distinguished from AChE from homozygous-susceptible and homozygous-resistant ticks. This is the first biochemical test for resistance to an acaricide. Rapid, sensitive tests like ours will allow resistance to organophosphates to be detected soon after it develops in the field, thus, the spread of resistance might be slowed and the useful life of acaricides extended.     

Prevalence of hard tick infestations in cattle of Mizoram,India

The present study was conducted to determine the prevalence of hard tick infestations in cattle of Mizoram from April 2017 to March 2018. The prevalence of hard tick infestations was studied in relation to sex and age of animals and seasonal changes in a year. Cattle of selected places were examined carefully for the presence of ticks and in positive cases ticks were collected manually and identified on the basis of morphological characters. A total of 713 cattle were examined and out of which 452 (63.39%) cattle were found to be infested with hard ticks and the prevalent tick species was Rhipicephalus (Boophilus) microplus. A significantly (p < 0.01) higher infestation was observed in female cattle (66.43%) than males (48.83%). Age-wise highest prevalence of tick infestations was found in 1–5 years (73.41%) age group followed by >5 years age group (61.92%) and < 1 year (42.65%) age group, respectively. Seasonally, the prevalence of hard tick infestation was found highest (p < 0.01) in monsoon (77.91%) and lowest in winter (43.16%). The present study provides data on tick infestation in cattle of Mizoram, India.     

Development of neonicotinoid resistance in the cotton aphid Aphis gossypii (Hemiptera: Aphididae) in Japan

A neonicotinoid-resistant Aphis gossypii Glover population was first detected from cucumber and sweet pepper crops in Miyazaki Prefecture, Japan, in April 2012. In this study, we determined the insecticide susceptibility of five field-collected populations of A. gossypii to seven neonicotinoids. Insecticide susceptibility was determined by performing bioassays and using the seedling-treatment method. Insecticide susceptibility of a susceptible population of A. gossypii, originally collected from a cucumber field in 2008, was also determined. High mortality (96.4–100 %) was observed in the susceptible population for the seven insecticides. In contrast, the mortality rate of the five field-collected populations collected in 2012 was low for five of the seven neonicotinoid insecticides tested: imidacloprid (26.7–65.5 %), dinotefuran (0–27.3 %), clothianidin (20.0–35.7 %), thiamethoxam (7.1–42.3 %), and nitenpyram (6.7–32.1 %). Mortality was 86.2–100 % for acetamiprid and 90.2–100 % for thiacloprid. The resistance ratio in comparison with the susceptible population was the highest for clothianidin (687-fold), lowest for thiacloprid (17-fold), and ranged from 43- to 253-fold for the other five neonicotinoids. Thus, the frequency of insecticide use needs to be reduced and other insecticides need to be employed in order to prevent the development of resistance to these two insecticides in the future.     

Detection of genetic variability in various isolates of cattle tick, <Emphasis Type="Italic">Boophilus microplus</Emphasis> from Tamil Nadu,India using PCR-RAPD analysis

The genomic DNA from ten isolates of the cattle tick, Boophilus microplus collected in and around Chennai, India, was analyzed by random amplified polymorphic DNA (RAPD) using PCR. Selected five random primers were used for the study of genetic variability among different isolates of B. microplus. A high degree of genetic polymorphism with a different pattern of RAPD profiles for each tick isolate was detected with all these random primers. This variability was also confirmed by similarity coefficient values and dendrogram which were performed using mean RAPD profiles for all the primers between various isolates of ticks. The findings suggest the existence of a complex genotypic diversity of the tick B. microplus in an endemic region such as Chennai.     

The effects of acaricide treatment of sheep on red grouse Lagopus lagopus scotica tick burdens and productivity in a multi‐host system

Ixodes ricinus (Ixodida: Ixodidae) ticks are of economic and pathogenic importance across Europe. Within the uplands of the U.K., management to reduce ticks is undertaken to benefit red grouse Lagopus lagopus scotica (Galliformes: Phasianidae). Management strategies focus on the acaricide treatment of domestic sheep Ovis aries (Artiodactyla: Bovidae), but the effectiveness of this is less certain in the presence of wild hosts, particularly red deer Cervus elaphus (Artiodactyla: Cervidae) and mountain hare Lepus timidus (Lagomorpha: Leporidae). This study examines the effects of sheep management on grouse tick burdens and productivity using sites with a range of wild host densities. Sites at which applications of acaricide were more frequent had lower tick burdens; this relationship was similar on sites with a range of deer densities. However, no direct link was detected between acaricide treatment interval and grouse productivity. Sites with higher deer densities had higher grouse tick burdens and lower productivity [mean ± standard error (SE) young : adult ratio: 1.2 ± 0.2] compared with sites with lower deer densities (mean ± SE young : adult ratio: 1.8 ± 0.1). Sites with higher grouse brood sizes and higher proportions of hens with broods were also those with higher mountain hare abundance indices. This study highlights the importance of the frequent treatment of sheep with acaricide to reduce tick burdens on grouse, even in the presence of wild hosts.     

Sheep tick Ixodes ricinus management on Welsh hill farms of designated conservation importance: implications for nationally declining birds

Impacts of sheep ticks Ixodes ricinus on livestock, gamebirds and wildlife are of concern across Europe. The present study describes livestock and tick management by 36 farmers from three upland sites of conservation importance in North Wales, where farmers consider that ticks have increased during the last 25 years. Sheep, average densities of 2.0 animals per ha were treated with pour‐on acaricides in spring, again in July, and also when removed from the moor in autumn. Given acaricide efficacy rates, sheep were susceptible to tick bites for half the period on the moor. Sheep from 17 farms were examined for ticks. Infestations were similar between farms and in relation to the acaricide used, averaging 9.3 ticks per sheep, although they were lower where the interval between successive acaricide treatments was shorter. Repeated sampling of sheep and red grouse chicks showed no annual difference in tick burdens on grouse chicks, which averaged 6.2 ticks per chick, although there were three‐fold fewer ticks on sheep in 2018 than in previous years. Tick bite rates on sheep and grouse were higher than elsewhere in the U.K. Most farmers interviewed would aim to improve their tick management using longer‐lasting acaricides and treating sheep more frequently, although they would need advice and financial help, which is currently unavailable via Government funded agri‐environment schemes.     

Influence of acaricide resistance on cattle-fever tick (Boophilus spp.) infestations in semi-arid thornshrublands: a simulation approach

Cattle-fever tick (Boophilus microplus and B. annulatus) populations that develop acaricide resistance become more difficult to control or eradicate. We used a simulation model to assess the direct and indirect effects of interactions among season, habitat type, grazing strategy, and acaricide resistance on the ability to eradicate Boophilus infestations in semi-arid thornshrublands of Texas, USA. Season of infestation appeared to have the strongest effect, with infestations begun on 27 September (autumn) tending to die out sooner than those begun on 1 March (spring) and to remain undetected. Habitat type had the next strongest effect, with infestations surviving much longer as canopy cover increased from uncanopied buffelgrass (Cenchrus ciliaris) habitats to mesquite (Prosopis glandulosa)-canopied grass habitats. Acaricide resistance had a moderate effect; as expected, highly resistant tick populations survived longer than those with no acaricide resistance. The importance of grazing strategy varied with changes in habitat type: as canopy cover increased, infestation duration increased faster under continuous grazing than under rotational grazing strategies. Importance of grazing strategy also varied with acaricide resistance: detected tick populations with no and slight acaricide resistance subjected to acaricide treatments tended to survive longer under rotational grazing than continuous grazing, due to reduced contact with a treated host. Populations with moderate and high resistance behaved more like untreated populations, tending to survive longer under continuous, rather than rotational, grazing, because they experienced less mortality on a treated host. Assuming acaricide treatments at 2-week intervals and maintenance of cattle in infested pastures, results indicate that, for each habitat type, infesting ticks have a threshold of acaricide resistance below which one can eradicate them faster with continuous grazing than with rotational grazing. As canopy cover increases, this threshold appears to shift from high resistance (in grass) to slight resistance (in mesquite).     

Antimicrobial Activities of Bacteriocins E 50–52 and B 602 Against Antibiotic-Resistant Strains Involved in Nosocomial Infections

The antimicrobial spectra of previously published bacteriocin E 50–52 (39 a.a.; 3,932 Da; pI = 8.5) and bacteriocin B 602 (29 a.a.; 3,864 Da; pI = 7.2) were determined. Named peptides were related to class IIa (pediocin-like) bacteriocins. Minimal inhibitory concentrations (MICs) of bacteriocins have been determined for bacterial isolates that were causative agents of nosocomial infections collected from Russian hospitals in 2003–2007, namely methicillin-resistant Staphylococcus aureus (MRSA) (n = 10); Acinetobacter baumannii (n = 11); Citrobacter freundii (n = 8); Escherichia coli (n = 9); Klebsiella pneumoniae (n = 10); Proteus spp. (n = 6); and Pseudomonas aeruginosa (n = 10). The majority of these tested isolates have been shown to be multidrug resistant and carry genetic determinants of antimicrobial resistance that were detected using polymerase chain reaction (PCR). The MICs of bacteriocin B 602 ranged from ≤0.025–1.56 μg/ml, and for bacteriocin E 50–52 from 0.05 to 6.25 μg/ml for all of 64 bacterial clinical isolates tested. Interestingly, the bacteriocins studied demonstrate activity on both Gram-positive and Gram-negative bacteria. Bacteriocins E 50–52 and B 602 show good activity against nosocomial bacterial agents resistant to many classes of modern antibacterials used in clinical practice. These bacteriocins should be examined as an alternative in treating infections caused by such agents.     

Abundance estimation of <Emphasis Type="Italic">Ixodes</Emphasis> ticks (Acari: Ixodidae) on roe deer (<Emphasis Type="Italic">Capreolus capreolus</Emphasis>)

Despite the importance of roe deer as a host for Ixodes ticks in central Europe, estimates of total tick burden on roe deer are not available to date. We aimed at providing (1) estimates of life stage and sex specific (larvae, nymphs, males and females, hereafter referred to as tick life stages) total Ixodes burden and (2) equations which can be used to predict the total life stage burden by counting the life stage on a selected body area. Within a period of 1½ years, we conducted whole body counts of ticks from 80 hunter-killed roe deer originating from a beech dominated forest area in central Germany. Averaged over the entire study period (winter 2007–summer 2009), the mean tick burden per roe deer was 64.5 (SE ± 10.6). Nymphs were the most numerous tick life stage per roe deer (23.9 ± 3.2), followed by females (21.4 ± 3.5), larvae (10.8 ± 4.2) and males (8.4 ± 1.5). The individual tick burden was highly aggregated (k = 0.46); levels of aggregation were highest in larvae (k = 0.08), followed by males (k = 0.40), females (k = 0.49) and nymphs (k = 0.71). To predict total life stage specific burdens based on counts on selected body parts, we provide linear equations. For estimating larvae abundance on the entire roe deer, counts can be restricted to the front legs. Tick counts restricted to the head are sufficient to estimate total nymph burden and counts on the neck are appropriate for estimating adult ticks (females and males). In order to estimate the combined tick burden, tick counts on the head can be used for extrapolation. The presented linear models are highly significant and explain 84.1, 77.3, 90.5, 91.3, and 65.3% (adjusted R ²) of the observed variance, respectively. Thus, these models offer a robust basis for rapid tick abundance assessment. This can be useful for studies aiming at estimating effects of abiotic and biotic factors on tick abundance, modelling tick population dynamics, modelling tick-borne pathogen transmission dynamics or assessing the efficacy of acaricides.     

Problems of isolating Borrelia burgdorferi from ticks collected in United Kingdom foci of Lyme disease

Abstract. Many isolates of Borrelia burgdorferi have been obtained from ticks and vertebrate tissues collected in North America and continental Europe but only one established culture of United Kingdom Borrelia burgdorferi has been recorded. In this paper we report the isolation of B.burgdorferi from one of 108 tick pools representing 733 ticks and eighty-four tissue samples from twenty-six rodents collected in the U.K., and the subsequent failure to establish the isolate (from ticks collected in Fordingbridge) in culture. In contrast, using identical techniques and culture medium, B.burgdorferi was isolated from one of seven tick pools collected in Switzerland, and from a single pool of ticks collected in Slovakia, and both isolates were successfully passaged. Analysis of questing I.ricinus collected from Fordingbridge by direct immunofluorescence showed 6/32 (19%) of adults and 8/108 (7%) of nymphs were positive for B. burgdorferi , although only one nymph contained ≥ 1000 spirochaetes. To examine further the problem of isolating U.K. B.burgdorferi , twelve Ixodes ricinus tick samples from Fordingbridge, a recognized focus of Lyme disease, were subjected to isolation and culturing techniques, and the procedures monitored by use of the polymerase chain reaction (PCR). Whereas 11/12 samples were PCR positive after 2 weeks in culture, only one was PCR positive after 4 weeks. Motile spirochaetes were not visible by dark-field microscopy in any of the cultures. The results indicate that the standard BSK II medium routinely used to isolate and culture B. burgdorferi does not readily support the replication of the Borrelia species endemic to the U.K.     

Prevalence of hard tick infestations in cattle of West Bengal,India

The present study was conducted to determine the prevalence of hard tick infestations in cattle of West Bengal from July 2015 to June 2016. The prevalence of hard tick infestations was studied in relation to sex and age of animals and seasonal changes in a year. Cattle of selected places were examined carefully for the presence of ticks and in positive cases ticks were collected manually and identified on the basis of morphological characters. A total of 310 cattle were examined and out of which, 130 (41.93%) cattle were found to be infested with hard ticks and the prevalent species were Rhipicephalus (Boophilus) sp., Hyalomma sp. and Haemaphysalis sp. of ticks. A significantly (p < 0.01) higher infestation was observed in female cattle (43.30%) than males (35.71%). Age-wise highest prevalence of tick infestations was found in <1 year (65%) age group followed by >3 years age group (36.8%) and 1–3 years (35.63%) age group, respectively. Seasonally, the prevalence of hard ticks was highest (p < 0.01) in monsoon (59.25%) and lowest in winter (27.09%). The study revealed that the prevalence of Rhipicephalus (Boophilus) sp. (32.25%) was significantly (p < 0.01) higher compared to Hyalomma (12.58%) and Haemaphysalis sp. (3.22%). The observations of the present study would provide a basis for evolving effective control strategy for the management of ticks in bovines of West Bengal.     

Detection of Rickettsia monacensis from Ixodes nipponensis collected from rodents in Gyeonggi and Gangwon Provinces, Republic of Korea

A total of 1,305 ticks were collected from wild rodents captured monthly, except July and August, during 2008 at three US-ROK operated military training sites and three US military installations in Gyeonggi and Gangwon Provinces, the Republic of Korea (ROK). Ixodes nipponensis was the most frequently collected tick (n = 1,299, 99.5 %), followed by Ixodes pomerantzevi (n = 6, 0.5 %). The ticks were pooled (1–15/sample) and tested by nested polymerase chain reaction (nPCR) for spotted fever group (SFG) rickettsiae with primer sets targeting the outer membrane protein B (ompB), citrate synthase (gltA), and 17-kDa antigen gene loci. A total of 115/197 (58.4 %) pools were positive by nPCR for the outer membrane protein ompB. Nucleotide sequence analysis of 105/115 (91.3 %) ompB targeted nPCR positive products showed a high degree of similarity to Rickettsia monacensis (99.3–100 %, n = 87) and R. japonica (99.5–100 %, n = 18). From the 87 positive samples demonstrating a high degree of similarity to R. monacensis, 15 were selected and analyzed by nPCR for gltA and the 17-kDa genes. A total of 12/15 pooled samples were positive for by nPCR for gltA, with amplicons demonstrating a high degree of similarity to R. monacensis (99.3–99.7 %). A total of 13/15 pooled samples were positive by nPCR for the 17-kDa gene, with amplicons demonstrating a high degree of similarity to R. monacensis (99.4–100 %). These findings demonstrate that R. monacensis is distributed throughout Gyeonggi and Gangwon Provinces in the ROK. Furthermore, data suggest a relative high prevalence of R. monacensis in the tick, I. nipponensis.     

Influence of tea tree oil (Melaleuca alternifolia) on the cattle tick Rhipicephalus microplus

The aim of this study was to verify the influence of tea tree oil (TTO) (Melaleuca alternifolia) tested in its pure and nanostructured (TTO nanoparticles) forms on the reproduction of female Rhipicephalus microplus. For our purpose, female ticks were collected from naturally infected animals and treated in vitro with TTO (1, 5, and 10 %) and TTO nanoparticles (0.075, 0.375, and 0.75 %). In order to validate the tests, they were performed in triplicate using positive (amitraz) and negative (untreated) controls. It was possible to observe that pure TTO (5 and 10 %) and TTO nanoparticles (0.375 and 0.75 %) showed 100 % reproductive inhibition on female ticks. Additionally, pure TTO (1 %) also showed an acaricide effect (70 %), similarly to the positive control (78.3 %). This is the first study demonstrating the activity of pure TTO and TTO nanoparticles on female ticks. Therefore, based on these results, we were able to show that both forms and all concentrations of M. alternifolia affected tick reproduction by inhibiting egg laying and hatching. We were also able to show that TTO nanoparticles potentiated the inhibitor effect of pure TTO on the reproduction of R. microplus.     

Investigating the loss of recruitment potential in red grouse (Lagopus lagopus scoticus): the relative importance of hen mortality,food supply,tick infestation and louping-ill

Ticks and their pathogens cause significant disease and economic loss in many animal populations. Despite this, experiments that test the impact of ticks and tick-borne diseases on wild animal populations are rare. Here, we report on an experiment assessing the effect of ticks on red grouse productivity and chick growth in relation to other causes of poor recruitment at two sites in the Scottish uplands during 2005. Treated hens received two leg bands impregnated with the acaricide permethrin, while controls hens were untreated. Chicks were captured at c.2 weeks of age and fitted with a metal patagial tag, and chicks from treated hens also received a permethrin-impregnated strip. Mean tick burdens in treated chicks were close to zero compared with a mean of around 12 in the control group. Although treatment reduced tick infestations, it did not increase brood size. Growth rates in chicks from control and treated hens were similar during the first 10 days and comparable with chicks fed an ad-lib invertebrate-based diet. These results suggest that in this case, neither ticks (and the tick transmitted louping-ill virus) nor food shortages was the main cause of chick mortality. However, mortality in the adult hens was around 35 %, and predation accounted for 62 % of these losses before broods fledged. Our results indicate that on our study sites, predation may have a more important impact on grouse population dynamics than ticks and tick-borne disease. We suggest that it may be more cost effective to determine the causes of poor grouse population performance before implementing popular but expensive tick control measures such as the culling of alternative hosts and running acaracide treated sheep ‘tick-mop’ flocks.     

Major surface protein 1a effects tick infection and transmission of Anaplasma marginale.

Anaplasma marginale, an ehrlichial pathogen of cattle and wild ruminants, is transmitted biologically by ticks. A developmental cycle of A. marginale occurs in a tick that begins in gut cells followed by infection of salivary glands, which are the site of transmission to cattle. Geographic isolates of A. marginale vary in their ability to be transmitted by ticks. In these experiments we studied transmission of two recent field isolates of A. marginale, an Oklahoma isolate from Wetumka, OK, and a Florida isolate from Okeechobee, FL, by two populations of Dermacentor variabilis males obtained from the same regions. The Florida and Oklahoma tick populations transmitted the Oklahoma isolate, while both tick populations failed to transmit the Florida isolate. Gut and salivary gland infections of A. marginale, as determined by quantitative PCR and microscopy, were detected in ticks exposed to the Oklahoma isolate, while these tissues were not infected in ticks exposed to the Florida isolate. An adhesion-recovery assay was used to study adhesion of the A. marginale major surface protein (MSP) 1a to gut cells from both tick populations and cultured tick cells. We demonstrated that recombinant Escherichia coli expressing Oklahoma MSP1a adhered to cultured and native D. variabilis gut cells, while recombinant E. coli expressing the Florida MSP1a were not adherent to either tick cell population. The MSP1a of the Florida isolate of A. marginale, therefore, was unable to mediate attachment to tick gut cells, thus inhibiting salivary gland infection and transmission to cattle. This is the first report of MSP1a being responsible for effecting infection and transmission of A. marginale by Dermacentor spp. ticks. The mechanism of tick infection and transmission of A. marginale is important in formulating control strategies and development of improved vaccines for anaplasmosis.     

Major surface protein 1a effects tick infection and transmission of Anaplasma marginale

Anaplasma marginale, an ehrlichial pathogen of cattle and wild ruminants, is transmitted biologically by ticks. A developmental cycle of A. marginale occurs in a tick that begins in gut cells followed by infection of salivary glands, which are the site of transmission to cattle. Geographic isolates of A. marginale vary in their ability to be transmitted by ticks. In these experiments we studied transmission of two recent field isolates of A. marginale, an Oklahoma isolate from Wetumka, OK, and a Florida isolate from Okeechobee, FL, by two populations of Dermacentor variabilis males obtained from the same regions. The Florida and Oklahoma tick populations transmitted the Oklahoma isolate, while both tick populations failed to transmit the Florida isolate. Gut and salivary gland infections of A. marginale, as determined by quantitative PCR and microscopy, were detected in ticks exposed to the Oklahoma isolate, while these tissues were not infected in ticks exposed to the Florida isolate. An adhesion-recovery assay was used to study adhesion of the A. marginale major surface protein (MSP) 1a to gut cells from both tick populations and cultured tick cells. We demonstrated that recombinant Escherichia coli expressing Oklahoma MSP1a adhered to cultured and native D. variabilis gut cells, while recombinant E. coli expressing the Florida MSP1a were not adherent to either tick cell population. The MSP1a of the Florida isolate of A. marginale, therefore, was unable to mediate attachment to tick gut cells, thus inhibiting salivary gland infection and transmission to cattle. This is the first report of MSP1a being responsible for effecting infection and transmission of A. marginale by Dermacentor spp. ticks. The mechanism of tick infection and transmission of A. marginale is important in formulating control strategies and development of improved vaccines for anaplasmosis.