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. 2012 Nov 15:5:263.
doi: 10.1186/1756-3305-5-263.

Semi-artificial mouse skin membrane feeding technique for adult tick, Haemaphysalis longicornis

Takeshi Hatta  1 , Takeharu Miyoshi, Makoto Matsubayashi, Md Khyrul Islam, M Abdul Alim, Anisuzzaman, Kayoko Yamaji, Kozo Fujisaki, Naotoshi Tsuji
Affiliations

Semi-artificial mouse skin membrane feeding technique for adult tick, Haemaphysalis longicornis

Takeshi Hatta et al. Parasit Vectors. .

Abstract

Background: An in vitro artificial feeding technique for hard ticks is quite useful for studying the tick-pathogen interactions. Here, we report a novel semi-artificial feeding technique for the adult parthenogenetic tick, Haemaphysalis longicornis, using mouse skin membrane.

Findings: Skin with attached adult ticks was removed from the mouse body at 4 to 5 days post-infestation for the construction of the feeding system. This system supplied with rabbit blood was kept in >95% relative humidity at 30°C during the feeding, and ticks were fully engorged (artificially engorged, AE) within 12 to 48 h. For comparison, ticks were fed to engorgement solely on rabbit or mouse for 5 days as controls (naturally engorged on rabbit, NEr, or mouse, NEm). Blood digestion-related gene expression in the midgut and reproductive fitness were compared. Body weight, egg mass weight, egg conversion ratio, and hatchability of eggs did not show any significant differences. We analyzed transcription profiles of selected genes assayed by quantitative RT-PCR and revealed similar patterns of expression between NEr and AE but some differences between NEm and AE or NEm and NEr.

Conclusions: Our results demonstrate that this semi-artificial feeding technique mimics natural feeding processes of ticks and can be utilized as a standardized method to inoculate pathogens, especially Babesia protozoa, into H. longicornis and possibly other tick species as well.

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Figures

Figure 1
Figure 1
Capitulum of H. longicornis. Scanning electron micrographs showing the mouthparts of H. longicornis from the dorsal aspect. Bar = 100 μm. h, hypostome; ch, chelicera; p, palps; bc, basis capituli; sc, scutum; l, leg.
Figure 2
Figure 2
Mouse-skin membrane-based semi-artificial feeding device. (A) Tick bite lesion of mouse skin used as a membrane in semi-artificial feeding technique. h, hypostome; ch, chelicera; ce, cement; en, endodermis; ep, epidermis; hy, hypodermis; fl, feeding lesion. (HE, Bar: 200 μm) (B) Lateral view of the semi-artificial feeding system with a feeding tick. (C) Schematic diagram of the system used in this study. (D-F) Chronological changes (0, 3, 5 h) of ticks feeding on the semi-artificial feeding system.
Figure 3
Figure 3
Phenotypic comparison of ticks fed on different feeding systems. (A) Engorged body weight of NEr (n = 6), AE (n = 4), and NEm (n = 6) ticks. The bars represent mean values and the error bars indicate the standard deviation. (B) Total egg mass weight measured at 10 DPO. (C) Egg conversion ratio (total egg weight/engorged body weight). (D) Comparison of hatchability of eggs (larval counts/total number of larvae and non-hatching eggs) derived from engorged ticks of different feeding systems. The results shown are from a single experiment and are representative of three independent experiments.
Figure 4
Figure 4
Comparison of transcriptional patterns of midgut-associated genes. Quantitative RT-PCR analysis of gene expression is presented relative to the internal standard β-actin and error bars represent the standard deviation. The results shown are from a single experiment and are representative of three independent experiments.
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