An alternative to conventional insect marking procedures: detection of a protein mark on pink bollworm by ELISA*

Rabbit immunoglobulin G (R-IgG) was used successfully as an external mark for thrips. Females of both Thrips tabaci Lindeman and Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae) were marked with 1 mg ml − 1 R-IgG solution with 1% 'Tween 20' by the contact exposure method. Determining the retention of the mark was by running the rinsing solution of individual thrips in an enzyme-linked immunosorbent assay (ELISA). The sandwich ELISA method was used with an additional biotin-avidin step. The threshold for a positive marking score was defined as three times the mean optical density readings of the negative control thrips. Under laboratory conditions, on bean pods, all marked thrips scored positive up to 6 days after marking (DAM). When marked thrips were kept in the laboratory on marigold flowers for 2 days, they all scored positive. When marked and unmarked thrips were placed together on these flowers, the mark was transferred to 10 -20% of the unmarked thrips and they became positive. Under field conditions, on sticky traps covered with water-base glue, 100, 80, and 20% of the marked T. tabaci scored positive by the 3rd, 6th, and 9th DAM, respectively. Under the same conditions 100, 90, and 10% of the marked F. occidentalis scored positive by the 3rd, 6th, and 9th DAM, respectively. The retention of the R-IgG decreased significantly under conditions of wetness and high humidity. After 6 days on chive plants kept at 80 -100% r.h., all marked thrips scored negative while on plants kept at 40 -60% r.h., 85% of the marked thrips scored positive. Rabbit IgG can be used as an external marker for thrips. The suitability of this marking method for dispersal studies of these important pests needs to be evaluated.

Section: Discussionsupporting

confidence: 77%

An immuno‐marking technique for thrips

Jasrotia

Ben‐Yakir²

2006

“…The darker gray bars at day 7 indicate the mean OD of the leaf tissue that Hippodamia convergens were exposed to during the fourth experiment (see the text for further details). A variety of methods (e.g., hand collection, sweepnet, sticky card, and fan trap) has been used to capture protein-marked insects for MRR-type studies with no apparent negative effect on the precision of the marking procedure (Hagler & Miller, 2002;Blackmer et al, 2004Blackmer et al, , 2006. by external contact from marked to unmarked individuals.…”

Section: Discussionmentioning

confidence: 99%

A protein‐based approach to mark arthropods for mark‐capture type research

Hagler

Jones

2010

Self Cite

A series of studies was conducted to test methods for marking a wide variety of arthropods with inexpensive proteins for mark-capture dispersal research. The markers tested included egg albumin protein in chicken egg whites and casein protein in bovine milk. The first study qualified the effectiveness of the two marks on more than 50 arthropod species inhabiting cotton via two application procedures. The application methods included: (1) a topical plus residue protein application, and (2) a residue-only protein application. Both protein marks, regardless of the method of application, were readily retained on the arthropod assemblage over the duration of the study. The second study determined how rapidly insects acquire chicken egg albumin protein after contact exposure to cotton tissue sprayed with an egg whites solution. Under laboratory conditions, the vast majority of adult Hippodamia convergens Guérin-Méneville (Coleoptera: Coccinellidae) and Lygus hesperus Knight (Heteroptera: Miridae) acquired the mark after 5 min, and immature Trichoplusia ni (Hübner) (Lepidoptera: Noctuidae) acquired the marker after 40 min. The third study determined how rapidly H. convergens and L. hesperus acquire bovine casein protein after contact exposure to either alfalfa, Medicago sativa L. (Fabaceae), or lesquerella, Lesquerella fendleri (Watson) (Brassicaceae), plants sprayed with a bovine milk solution. These insects rapidly acquired the casein mark from the plant residue under field conditions. A final study determined how long H. convergens retain casein protein after 24-h exposure to alfalfa and lesquerella plants containing a 7-day-old residue of bovine milk. Approximately 95% of the H. convergens maintained the casein mark for 2 days after removal from each type of plant.

“…However, the relative contribution of internal vs. external labeling was not evaluated in that study. In a different study with P. gossypiella , an external label of rabbit IgG (5 mg ml −1 ) was transferred from externally marked pupae to adults (Hagler & Miller, 2002). The egg labeling experiment in the current study revealed that rabbit or chicken IgG label is well‐retained throughout the egg stage, thus allowing either label to be utilized in future studies on egg predation, as described by Mansfield et al.…”

Section: Discussionmentioning

confidence: 99%

Retention of immunolabels by Diorhabda carinulata, a biological control agent of saltcedar

Williams¹,

Hagler²,

Tonkel³

2011

Self Cite

Marking biological control agents facilitates studies of dispersal and predation. This study examines the feasibility of marking the various life stages of a weed biological control agent, Diorhabda carinulata (Desbrochers) (Coleoptera: Chrysomelidae), by submersion in rabbit or chicken immunoglobulin G (IgG) protein solutions. We determined whether externally applied IgGs are effective labels of the various lifestages, whether IgGs can be retained between D. carinulata lifestages, and to what extent abiotic factors associated with field conditions mediate label retention. The presence of the labels on the various lifestages of the beetles was detected by IgG‐specific enzyme‐linked immunosorbent assays. Duration of each immunolabel was measured on eggs and larvae in laboratory studies and on adults in laboratory and field studies. For adults, both labels showed high (>80%) retention for ca. 14 days after marking under field and laboratory conditions. Temperature and type of label (rabbit or chicken) had only a minimal effect on marker retention. Externally marked eggs exhibited high (100%) retention for both proteins over the entire duration of the egg stage. Interestingly, some larvae emerging from externally labeled eggs contained both external and internal IgG marks. To our knowledge, this is the first case of an IgG being transferred from the egg to larva of an insect. Age of eggs at the time of label application affected the intensity of the external label on neonates. For instance, larvae that emerged from eggs that were >1 day old when labeled exhibited stronger label retention than larvae that emerged from eggs that were 1 day old when labeled. For larvae, retention of rabbit IgG was greater than retention of chicken IgG. Label retention declined as larvae aged; larvae >3 days old retained significantly less label than did neonate larvae. Both IgG labels were retained from the first to second instar, but at a very low rate of <10%. Overall, our study demonstrates that protein‐marking technology has potential for use in studies of dispersal and predator–prey associations for D. carinulata.