Bed bugs from a laboratory colony (Cincinnati strain, Sierra Research Laboratories Inc., established 2007) reared on commercially obtained rabbit blood (Hemostat Laboratories, Dixon, CA) using a membrane feeding system (Hemotek Ltd. Blackburn, UK) were examined as negative controls. Also included was a set of positive controls consisting of bed bugs fed blood from wild-type C57BL/6 laboratory mice. This blood was freshly extracted from necropsied mice and immediately administered using the membrane feeding system (Hemotek Ltd.). Both groups of control bed bugs were maintained in an incubator at 28 ± 1 °C and 60–70% relative humidity on a 12:12-h photoperiod.
Membrane feeding system
Manufactured by Hemotek
Sourced in United Kingdom, Sweden, France
The Membrane Feeding System is a laboratory equipment designed to provide a controlled environment for the feeding and study of insects and other small organisms. The core function of this system is to facilitate the delivery of a liquid diet or other substances through a membrane, allowing for the observation and analysis of the feeding behavior and physiological responses of the target organisms.
Automatically generated - may contain errors
Lab products found in correlation
Human blood, by Hemotek (2 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (1 mentions)
Pellet pestle, by Merck Group (1 mentions)
C6 36 cells, by American Type Culture Collection (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Prism 9, by GraphPad (1 mentions)
Nanoject 2, by Drummond (1 mentions)
Sheep blood, by Hemotek (1 mentions)
Defibrinated sheep blood, by Solarbio (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Parafilm, by Merck Group (1 mentions)
Smz745, by Nikon (1 mentions)
Parafilm, by Amcor (1 mentions)
72 protocols using membrane feeding system
1
Bed Bug Control Assay Protocol
2
Chikungunya Virus Infection in Aedes albopictus
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Laboratory colonies of A. albopictus were established from field collections in 2011 in Phu Hoa, Ben Cat District, Binh Duong Province, Vietnam. All the experiments were performed within 19 generations of laboratory colonization. The insectary conditions for mosquito maintenance were 28°C, 70% relative humidity, and a 12-h light and 12-h dark cycle. Adults were maintained with permanent access to 10% sucrose solution. Adult females were offered commercial rabbit blood (BCL) twice a week through a membrane feeding system (Hemotek Ltd.). 6–8 days old female mosquitoes were fed with 105 PFU of CHIKV diluted in preached human blood (iCareB platform, Institut Pasteur). Mosquitoes were offered the infectious or control blood meal for 30 min through a membrane feeding system (Hemotek Ltd) set at 37°C with a piece of desalted pig intestine as the membrane. Following the blood meal, fully engorged females were selected and incubated for 2 and 8 days at 28°C, 70% relative humidity and under a 12 h light: 12 h dark cycle with permanent access to 10% sucrose. Viral titers in individual mosquitoes were determined by plaque assay. Also, total RNA was extracted with TriZol reagent and the composition of the 3’UTR in the population was analyzed as described above.
3
Rearing and Maintaining Aedes aegypti Mosquitoes
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Ae. aegypti mosquitoes Liverpool strain were reared at 28 ± 1°C, 80% humidity with a 12h:12h light:dark cycle. Eggs were hatched in tap water, and larvae were fed with fish food powder (Tetramin) every other day. Adults were allowed constant access to 10% (w/v) sucrose in water, and females were fed on human blood (Sanquin Blood Supply Foundation) through a membrane feeding system (Hemotek Ltd.). For injection experiments, females were separated, offered a blood meal, and allowed to lay eggs 3 to 4 d later by providing a moist surface and placing the mosquitoes in the dark. Embryos were collected at the indicated time points after a 30-min egg laying period. For Figure 7 A,B, Ae. aegypti Jane mosquitoes were used as described previously (Halbach et al. 2020 (link)).
4
Mosquito Infection and Diapause Study
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Prior to offering them an infectious meal, 5–7-day-old female mosquitoes were deprived of sucrose solution for 16 h. The infectious blood meal was a mixture of fresh virus suspension and mouse blood at a ratio of 1:1. While feeding, the viral blood meal was constantly warmed to 37°C using a Hemotek membrane feeding system. After 30 min exposure to the blood meal, mosquitoes were CO2 anesthetized and only fully engorge females (over 500 individuals per cage) were transferred immediately to cages. The LD females were daily given 10% sucrose in a chamber at 28°C and 80% RH under LD photoperiods. In contrast, the females under the diapause-inducing SD photoperiod were kept in the same environment. Besides, uninfected control cages under LD and SD conditions were fed with the uninfected blood meal consisting of 1:1 mouse blood and Dulbecco's modified Eagle's medium (Gibco?; Invitrogen, Beijing, China), which were treated in parallel to the infectious blood-fed cages.
5
Mosquito Feeding on Rabbit Blood
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Boxes of 60 one-week-old female adults were fed for 15 min through a pig intestine membrane covering the base of a feeder (Hemotek membrane feeding system, UK) containing 1.4 mL of rabbit erythrocytes supplemented with 10 mM adenosine triphosphate (ATP) as a phagostimulant and 0.7 mL of viral stock to obtain a final titer of 106.5 and 107.5 ffu/mL. Only fully engorged mosquitoes were kept and maintained in containers placed in climatic chambers at 28°C ± 0·1°C until processing at different days post-infection (dpi). Mosquitoes were fed with 10% sucrose solution.
6
Evaluating Irradiation Effects on Mosquito Reproduction
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For each dose, 20 irradiated females and 20 virgin non-irradiated males (both 3 days post-emergence) were selected at random and placed together in cages (30x30x30 cm) with continuous access to 10% sucrose solution. At 4 days post-emergence, females were offered a sheep's blood meal once a day for three consecutive days, using a Hemotek membrane feeding system. After this time each female was placed individually in a 50 ml plastic centrifuge tube containing 10 ml of deionized water and a strip of filter paper (9 x 4 cm) as an oviposition substrate. The tube was sealed with a ventilated mesh lid. The total number of eggs laid by each female (fecundity) and the number of eggs that hatched out of the total number of eggs produced per female (fertility) were determined over a single gonotrophic cycle. To determine the effects of irradiation on males, the experiment was repeated using 20 irradiated males and 20 virgin females that had not been irradiated. Controls consisted of groups of 20 males and 20 females that had not been irradiated. Three replicates (cages) were performed for each treatment.
7
Ae. aegypti Mosquito Superinfection Assay
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Two rounds of experimental infection assays were conducted, using the same protocol for mosquito superinfection. Thirty-six hours before infection, 6–7 days old inseminated Ae. aegypti females from each of the three populations (Barra, Deodoro, and Porto) were separated in 18 cylindrical plastic cages (70 mosquitoes/cage) for blood feeding. Sugar supply was removed 36-h before mosquitoes were challenged with the infective blood meal to increase female’s avidity. The oral infection procedures were performed through a membrane feeding system (Hemotek, Great Harwood, UK), adapted with a pig-gut covering, which gives access to the defibrinated rabbit blood. The infective blood meals consisted of 1 ml of supernatant of infected cell culture, 2 ml of washed rabbit erythrocytes, and 0.5 mM of ATP as phagostimulant. The same procedure and membrane feeding apparatus were used to feed control mosquitoes, but they received a noninfectious blood meal, with 1 ml of cell culture medium replacing the viral supernatant.
8
Mosquito Infection with Zika Virus
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Seven-day-old female mosquitoes (n>500) were starved for 12 h prior to the infectious blood meal. The blood meal consisted of 1:1 mouse blood and virus suspension. Oral infections with ZIKV were performed with virus at a titer of 3 × 105 plaque-forming units/mL, verified by titration in a standard plaque assay.27 (link) The mosquitoes were fed with an infectious blood meal that was warmed to 37 °C using a Hemotek membrane feeding system housed in a feeding chamber. After 30 min, the mosquitoes were cold anesthetized, and fully engorged females were transferred to 300 mL ca. plastic cups, which were maintained at 29±1 °C, 75±5% relative humidity, a 14 h/10 h light:dark photoperiod and 8% sugar water.
9
Rearing and Propagation of An. coluzzii Mosquitoes
10
Rearing and Propagation of An. coluzzii Mosquitoes
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An. coluzzii adults were reared at 27°C, 75% humidity under a 12 h light/12 h dark photoperiod and supplied with 10% sucrose water in the Vanderbilt University Insectary (Fox et al., 2001 (link); Suh et al., 2016 (link)). For stock propagation, 5- to 7-day-old mated females were blood-fed for 30–45 min using a membrane feeding system (Hemotek, Lancaster, UK) filled with defibrinated sheep blood purchased from Hemostat Laboratories (Dixon, CA, USA).
Mosquito larvae were reared in distilled water at 27°C under the standard 12 h light/12 h dark cycle, with approximately 300 larvae per rearing pan in 1 L H2O. The larval food was made from 0.12 g/mL Kaytee Koi’s Choice premium fish food (Chilton, WI, US) plus 0.06 g/mL yeast in distilled water and subsequently incubated at 4°C overnight for fermentation. For first and second instar larvae, 0.08 mL larval food was added into the water every 24 h. For third and fourth instar larvae, 1 mL larval food was added.
Mosquito larvae were reared in distilled water at 27°C under the standard 12 h light/12 h dark cycle, with approximately 300 larvae per rearing pan in 1 L H2O. The larval food was made from 0.12 g/mL Kaytee Koi’s Choice premium fish food (Chilton, WI, US) plus 0.06 g/mL yeast in distilled water and subsequently incubated at 4°C overnight for fermentation. For first and second instar larvae, 0.08 mL larval food was added into the water every 24 h. For third and fourth instar larvae, 1 mL larval food was added.
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