In vitro feeding of instars of the ixodid tick Amblyomma variegatum on skin membranes and its application to the transmission of Theileria mutans and Cowdria ruminantium

Abstract

An in vitro feeding method using rabbit or cattle skin membranes, applied successfully to all stages (larvae, nymphae and adults) of the ioxodid tick, Amblyomma variegatum, is described. The feeding apparatus consisted of a blood container with a membrane placed on top of a tick containment unit. A carbon dioxide atmosphere of between 5 and 10% and a temperature of 37 degrees C were used as stimulants for the attachment of the ticks. High CO2 concentrations in the atmosphere improved the feeding success of all instars. The effect of anticoagulation methods for the bloodmeal was investigated, and heparinized blood was found to be the most suitable for tick feeding. When the bloodmeal was replaced by tissue culture medium for feeding nymphs the subsequent moulting success was reduced. Adult ticks of both sexes remained attached for up to 16 days, until completion of their bloodmeals. All stages of the tick fed on whole blood in the artificial feeding system and all reached engorged weights less than those achieved by control ticks fed on experimental animals. A large proportion of ticks, fed artificially on whole blood, moulted or laid eggs successfully. The method was successfully applied for the transmission of Theileria mutans and Cowdria ruminantium to cattle.

Establishment of Boophilus microplus Infected with Babesia bigemina by Using in vitro Tube Feeding Technique.

“…The tube feeding technique need pre-fed ticks, so at least one cattle was required to supply the pre-fed female ticks. Using the membrane system, that is another in vitro feeding system [5,6,9,[12][13][14][15][16], the more numbers of experimental cattle could be saved. The second advantage of the technique is that the relationship between tick and protozoa can be studied quantitavely.…”

“…To save the numbers of experimental animals and study the relationship between ticks and tick-borne protozoa more directly, many in vitro feeding techniques have been reported [9,12,[14][15][16] since the first method had attempted [1]. B. microplus can also feed host blood through both membranes [5,6,13] and capillary tubes [17].…”

Laboratory artificial infection of hard ticks: a tool for the analysis of tick-borne pathogen transmission

“…Indeed, rabbits are classically used to feed ticks in laboratory: immature stages of Rhipicephalus evertsi evertsi (Londt and Van der Bijl 1977), all life stages of I. scapularis, I. pacificus, A. americanum, Dermacentor occidentalis, D. variabilis, Haemaphysalis leporispalustris and R. sanguineus (Troughton and Levin 2007), R. appendiculatus (Bailey 1960), A. variegatum (Voigt et al 1993), D. andersoni (Howarth and Hokama 1983), A. hebraeum (Heyne et al 1987), I. ricinus (Bonnet et al 2007). In these cases, the typical way to engorge ticks is to use feeding bags or capsules glued to clean-shaven skin on the back of the animal (Figure 1).…”

“…Since then, several membranes from different animal origins have been used with variable success to engorge ticks, including pieces of cattle skin for R. microplus (Kemp et al 1975) and A. variegatum FIGURE 2: View of attached I. ricinus nymphs on a rabbit skin used in the membrane feeding system. (Voigt et al 1993;Young et al 1996), calf mesentery and modified Baudruche membranes for R. microplus (Kemp et al 1975;Waladde et al 1979) and R. appendiculatus (Waladde et al 1991;Young et al 1996), rabbit skin for A. variegatum (Voigt et al 1993;Young et al 1996), D. andersoni (Howarth and Hokama 1983), R. appendiculatus (Musyoki et al 2004) and I. ricinus (Bonnet et al 2007) (Figure 2), mouse skin for D. andersoni (Howarth and Hokama 1983;Paine et al 1983) and I. scapularis (Burkot et al 2001), and gerbil skin for I. ricinus (Bonnet et al 2007). Membranes of non-animal origin made from silicone have also been used with success, particularly for feeding the different instars of A. hebraeum (Kuhnert et al 1995), I. ricinus females (Krober and Guerin 2007b), A. cajennense adults (de Moura et al 1997), and recently H. anatolicum anatolicum and H. dromedarii (Tajeri and Razmi 2011).…”

“…Membranes of non-animal origin made from silicone have also been used with success, particularly for feeding the different instars of A. hebraeum (Kuhnert et al 1995), I. ricinus females (Krober and Guerin 2007b), A. cajennense adults (de Moura et al 1997), and recently H. anatolicum anatolicum and H. dromedarii (Tajeri and Razmi 2011). However, without the addition of specific stimuli, the use of such membranes has proved ineffective for ticks such as A. variegatum (Voigt et al 1993). This is related to the fact that one of the greatest difficulties is to encourage the attachment of unfed ticks (see above).…”