Three-Dimensional Culture of Rhipicephalus (Boophilus) microplus BmVIII-SCC Cells on Multiple Synthetic Scaffold Systems and in Rotating Bioreactors

Tick cell culture facilitates research on the biology of ticks and their role as vectors of pathogens that affect humans, domestic animals, and wildlife. Because two-dimensional cell culture doesn’t promote the development of multicellular tissue-like composites, we hypothesized that culturing tick cells in a three-dimensional (3-D) configuration would form spheroids or tissue-like organoids. In this study, the cell line BmVIII-SCC obtained from the cattle fever tick, Rhipicephalus (Boophilus) microplus (Canestrini, 1888), was cultured in different synthetic scaffold systems. Growth of the tick cells on macrogelatinous beads in rotating continuous culture system bioreactors enabled cellular attachment, organization, and development into spheroid-like aggregates, with evidence of tight cellular junctions between adjacent cells and secretion of an extracellular matrix. At least three cell morphologies were identified within the aggregates: fibroblast-like cells, small endothelial-like cells, and larger cells exhibiting multiple cytoplasmic endosomes and granular vesicles. These observations suggest that BmVIII-SCC cells adapted to 3-D culture retain pluripotency. Additional studies involving genomic analyses are needed to determine if BmVIII-SCC cells in 3-D culture mimic tick organs. Applications of 3-D culture to cattle fever tick research are discussed.

Biosurveillance and Research Needs Involving Area-Wide Systematic Active Sampling to Enhance Integrated Cattle Fever Tick (Ixodida: Ixodidae) Eradication

The one-host cattle fever tick, Rhipicephalus (Boophilus) annulatus (Say), and southern cattle fever tick, Rhipicephalus (Boophilus) microplus (Canestrini), are important ectoparasitic pests of cattle, Bos taurus L., mostly for transmitting the causal agents of bovine babesiosis. Bovine babesiosis inflicted substantial cattle production losses in the United States before the vectors were eliminated by 1943, with the exception of a Permanent Quarantine Zone in South Texas, a buffer along the Mexico border where the invasive ixodids remain. As suitable hosts, infested white-tailed deer and nilgai antelope populations disperse R. annulatus and R. microplus, which increases the risk for emergence of bovine babesiosis in the United States. A R. microplus incursion first detected in 2016 on the South Texas coastal plain wildlife corridor involved infestations on cattle, nilgai antelope, white-tailed deer, and vegetation. Efforts at passive sampling of Rhipicephalus (Boophilus) spp. on hosts are concentrated in the Permanent Quarantine Zone. Hence, a knowledge gap exists on the full extent of the recent incursions. Area-wide, systematic, active sampling and supportive research, involving the Permanent Quarantine Zone, Temporary Quarantine Zone, most of the coastal plain, and other parts of Texas outside of the quarantine zones, are needed to bridge the knowledge gap. Herein, we provide research perspectives and rationale to develop and implement systematic active sampling that will provide an increasingly accurate assessment of Rhipicephalus (Boophilus) spp. distribution in Texas. We suggest that this is essential to advance integrated vector-borne animal disease eradication approaches for keeping cattle free of bovine babesiosis.

Effects of white-tailed deer habitat use preferences on southern cattle fever tick eradication: simulating impact on “pasture vacation” strategies

Background Rhipicephalus (Boophilus) annulatus and Rhipicephalus (Boophilus) microplus (southern cattle fever tick; SCFT), collectively known as cattle-fever ticks (CFTs), are vectors of protozoal parasites (Babesia bigemina and Babesia bovis) that cause bovine babesiosis (also known as cattle fever). One traditional strategy for CFT eradication involves the implementation of a “pasture vacation,” which involves removing cattle (Bos taurus) from an infested pasture for an extended period of time. However, vacated pastures are often inhabited by wildlife hosts, such as white-tailed deer (WTD; Odocoileus virginianus), which can serve as alternate hosts for questing CFTs. We hypothesized that the distribution of host-seeking larvae among habitat types post-pasture vacation would reflect habitat use patterns of WTD, and in turn, affect the subsequent rate of pasture infestation by CFT. Methods We adapted a spatially explicit, individual-based model to simulate interactions among SCFT, cattle, and WTD as a tool to investigate the potential effects of WTD habitat use preferences on the efficacy of a pasture vacation. We parameterized the model to represent conditions typical of rangelands in south Texas, USA, simulated a 1-year pasture vacation under different assumptions regarding WTD habitat use preferences, and summarized effects on efficacy through (1) time post-vacation to reach 100% of pre-vacation densities of host-seeking larvae, and (2) the ecological conditions that resulted in the lowest host-seeking larval densities following pasture vacation. Results Larval densities at the landscape scale varied seasonally in a similar manner over the entire simulation period, regardless of WTD habitat use preferences. Following the removal of cattle, larval densities declined sharply to < 100 larvae/ha. Following the return of cattle, larval densities increased to > 60% of pre-vacation densities ≈ 21 weeks post-vacation, and reached pre-vacation levels in less than a year. Trends in larval densities in different habitat types paralleled those at the landscape scale over the entire simulation period, but differed quantitatively from one another during the pasture vacation. Relative larval densities (highest to lowest) shifted from (1) wood/shrub, (2) grass, (3) mixed-brush during the pre-vacation period to (1) mixed-brush, (2) wood/shrub, (3) grass or (1) wood/shrub, (2) mixed-brush, (3) grass during the post-vacation period, depending on WTD habitat use preferences. Conclusions By monitoring WTD-driven shifts in distributions of SCFT host-seeking larvae among habitat types during simulated pasture vacation experiments, we were able to identify potential SCFT refugia from which recrudescence of infestations could originate. Such information could inform timely applications of acaricides to specific refugia habitats immediately prior to the termination of pasture vacations.

Enhanced biosurveillance of high-consequence invasive pests: southern cattle fever ticks, Rhipicephalus (Boophilus) microplus, on livestock and wildlife

Background Some tick species are invasive and of high consequence to public and veterinary health. Socioeconomic development of rural parts of the USA was enabled partly through the eradication by 1943 of cattle fever ticks (CFT, Rhipicephalus (Boophilus) annulatus and R. (B.) microplus). The southern cattle fever ticks (SCFT, R. (B.) microplus) remain a real and present threat to the USA animal agriculture because they are established in Mexico. Livestock-wildlife interactions in the Permanent Quarantine Zone (PQZ) established by the century-old Cattle Fever Tick Eradication Programme (CFTEP) in south Texas endanger its operations. Methods We describe a spatially-explicit, individual-based model that simulates interactions between cattle, white-tailed deer (WTD, Odocoileus virginianus), and nilgai (Boselaphus tragocamelus) to assess the risk for SCFT infestations across the pathogenic landscape in the PQZ and beyond. We also investigate the potential role of nilgai in sustaining SCFT populations by simulating various hypothetical infestation and eradication scenarios. Results All infestation scenarios resulted in a phase transition from a relatively small proportion of the ranch infested to almost the entire ranch infested coinciding with the typical period of autumn increases in off-host tick larvae. Results of eradication scenarios suggest that elimination of all on-host ticks on cattle, WTD, or nilgai would have virtually no effect on the proportion of the ranch infested or on the proportions of different tick habitat types infested; the entire ranch would remain infested. If all on-host ticks were eliminated on cattle and WTD, WTD and nilgai, or cattle and nilgai, the proportions of the ranch infested occasionally would drop to 0.6, 0.6 and 0.2, respectively. Differences in proportions of the ranch infested from year to year were due to primarily to differences in winter weather conditions, whereas infestation differences among tick habitat types were due primarily to habitat use preferences of hosts. Conclusions Infestations in nilgai augment SCFT refugia enabled by WTD and promote pest persistence across the landscape and cattle parasitism. Our study documented the utility of enhanced biosurveillance using simulation tools to mitigate risk and enhance operations of area-wide tick management programmes like the CFTEP through integrated tactics for SCFT suppression.

Landscape Ecology of Rhipicephalus (Boophilus) microplus (Ixodida: Ixodidae) Outbreaks in the South Texas Coastal Plain Wildlife Corridor Including Man-Made Barriers

Landscape features and the ecology of suitable hosts influence the phenology of invasive tick species. The southern cattle fever tick, Rhipicephalus (Boophilus) microplus (Canestrini) (Ixodida: Ixodidae), vectors causal agents of babesiosis in cattle and it infests exotic, feral nilgai, Bosephalus tragocamelus Pallas, and indigenous white-tailed deer, Odocoilus virginianus (Zimmerman), on the South Texas coastal plain wildlife corridor. The corridor extends from the Mexico border to cattle ranches extending north from inside Willacy Co. Outbreaks of R. microplus infesting cattle and nondomesticated ungulate hosts since 2014 in the wildlife corridor have focused attention on host infestation management and, by extension, dispersal. However, there is a knowledge gap on the ecology of R. microplus outbreaks in the South Texas coastal plain wildlife corridor. Ixodid distribution on the wildlife corridor is strongly influenced by habitat salinity. Saline habitats, which constitute ≈25% of the wildlife corridor, harbor few ixodids because of occasional salt toxicity from hypersaline wind tides and infrequent storm surges, and from efficient egg predation by mud flat fiddler crabs, Uca rapax (Smith). Rhipicephalus microplus infestations on nilgai were more prevalent in part of the corridor with mixed low salinity and saline areas than in an area that is more extensively saline. The different levels of R. microplus infestation suggest that man-made barriers have created isolated areas where the ecology of R. microplus outbreaks involve infested nilgai. The possible utility of man-made barriers for R. microplus eradication in the lower part of the South Texas coastal plain wildlife corridor is discussed.

Lethal Effects of a Commercial Diatomaceous Earth Dust Product on Amblyomma americanum (Ixodida: Ixodidae) Larvae and Nymphs

With increasing development of resistance to conventional synthetic acaricides in economically and medically important ixodid species, interest in finding alternative control tactics has intensified. Laboratory bioassays were conducted, using the lone star tick, Amblyomma americanum (L.), as a model species, to assess the efficacy of a diatomaceous earth-based product, Deadzone, in comparison with a silica gel-based product, CimeXa. CimeXa is already known to be highly lethal against A. americanum larvae and nymphs. The two dust treatments were 100% effective against larvae and nymphs within 24 h after contact occurred by immersion in dry dusts and after crawling across a surface treated with the dry dusts. Contact by crawling on a dried aqueous film of the dusts, even at a concentration of 10%, was not as effective as exposure to the dusts in dry powder form. As has been demonstrated with CimeXa, it is likely that Deadzone will be capable of providing prophylactic protection of cattle from economically important one-host ixodids, such as the southern cattle fever tick, Rhipicephalus microplus (Canestrini), which vectors the causal agents of babesiosis. Diatomaceous earth can be stored indefinitely, will remain efficacious for as long as sufficient quantities remain on the substrate, it is a natural (organic) substance, and it might be amenable for limited use in environmentally protected habitats.

Lethal Effects of a Silica Gel + Thyme Oil (EcoVia) Dust and Aqueous Suspensions on Amblyomma americanum (Ixodida: Ixodidae) Larvae and Nymphs

Ixodids suck blood from host animals and transmit pathogens that cause important medical and veterinary diseases. As synthetic conventional acaricide resistance becomes increasingly common, alternative tactics are likely to enhance control efforts. Laboratory bioassays assessed the lethality of CimeXa, a commercial silica gel desiccant dust, and EcoVia, a commercial product containing silica gel + thyme oil, against lone star tick, Amblyomma americanum (L.), larvae and nymphs. Both life stages were completely killed by CimeXa by 24 h, and EcoVia achieved total larval mortality within 1 h when they were initially immersed in the dusts and when they crawled across treated substrate. Larvae were killed faster than nymphs. Temporary immersion of A. americanum larvae and nymphs in aqueous suspensions of the dusts were less effective than exposure to the dusts applied dry. Larval and nymphal mortality associated with crawling on dried aqueous suspensions of the products was also not as strong as when the immature life stages crawled across substrate treated with dry dusts, and EcoVia was not more efficient than CimeXa. CimeXa and EcoVia can likely be used to protect cattle in a prophylactic context because silica gel does not degrade, and EcoVia might be effective at eliminating ticks that are feeding on cattle as well. We suggest that the two dust products be assessed for efficacy against other ixodid species, such as the southern cattle fever tick, Rhipicephalus microplus (Canestrini), which transmits the causal agents of babesiosis to cattle.