BackgroundThe development of standardized in vitro feeding methods for ixodid ticks has been hampered by their complex feeding behaviour and the long duration of their blood meal. In this study, we aimed to optimize several parameters for the in vitro feeding of adult Dermacentor reticulatus.MethodsTicks were fed on heparinized bovine blood collected at a slaughterhouse, using a modified silicone membrane feeding assay. Effects on tick feeding and fecundity of different blood meal treatments (freezing, irradiation, addition of antibiotics), ambient conditions (increased CO2 concentration) and phagostimulant use (addition of 2 g/l and 4 g/l glucose to the blood meal) were systematically evaluated.ResultsAlthough fungal growth occurred more frequent in feeding units of ticks fed on defrosted blood, the attachment rate, engorgement mass and fecundity of females fed on defrosted blood did not significantly differ from that of ticks fed on fresh blood. A reduction in the fecundity of female D. reticulatus ticks was observed when ticks were fed with gamma-irradiated blood or untreated blood compared to blood treated with gentamycin. Both the engorgement mass and fecundity increased when ticks were fed at a 5% CO2 level. A non-significant increase in the engorgement mass and engorgement rate of D. reticulatus was observed when blood was supplemented with 4 g glucose per litre compared to 2 g/l.ConclusionAn artificial feeding method was adapted for the feeding of adult D. reticulatus ticks. Of all parameters tested, only the artificial feeding at 5% CO2 levels resulted in a significant increase (P < 0.05) in the engorgement mass and fecundity of female D. reticulatus ticks. The supplementation of blood with antibiotics resulted in a significantly higher tick fecundity in comparison to ticks fed with untreated or irradiated blood.
The long feeding duration of ixodid ticks and need for regular blood changes turns the artificial feeding of ticks into a tedious process. To reduce the number of blood changes, a semi-automated system (SAS) for the artificial feeding of hard ticks was developed and evaluated. It consisted of a glass feeding reservoir that can accommodate six tick feeding chambers. A peristaltic pump was used to pump blood through the feeding reservoir, which was changed once daily. Groups of Dermacentor reticulatus and Ixodes ricinus adults were fed simultaneously in both the SAS and a conventional in vitro feeding system. In the conventional system, feeding chambers were hung inside a glass beaker filled with blood that was replaced twice daily. Dermacentor reticulatus adults fed in the SAS obtained significantly higher engorgement weights. Although engorgement rates between both systems were comparable, significantly more SAS-fed females laid fertile egg batches. The egg batch weight of SAS-fed females was also significantly higher. In contrast, the engorgement rate and fecundity of SAS-fed I. ricinus were significantly reduced in comparison to ticks fed in the conventional system. This reduction was likely to be caused by fungal infestation, which could spread between feeding chambers in the SAS. Although the SAS reduced the workload compared to the conventional feeding system and showed promising results for the in vitro feeding of D. reticulatus adults, measures to prevent fungal infestations in the SAS should be considered in future studies.
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