scholarly journals Cryopreservation of Anopheles stephensi embryos

2022 ◽  
Vol 12 (1) ◽  
Author(s):  
Eric R. James ◽  
Yingda Wen ◽  
James Overby ◽  
Kristen Pluchino ◽  
Shane McTighe ◽  
...  

AbstractThe ability to cryopreserve mosquitoes would revolutionize work on these vectors of major human infectious diseases by conserving stocks, new isolates, lab-bred strains, and transgenic lines that currently require continuous life cycle maintenance. Efforts over several decades to develop a method for cryopreservation have, until now, been fruitless: we describe here a method for the cryopreservation of Anopheles stephensi embryos yielding hatch rates of ~ 25%, stable for > 5 years. Hatched larvae developed into fertile, fecund adults and blood-fed females, produced fully viable second generation eggs, that could be infected with Plasmodium falciparum at high intensities. The key components of the cryopreservation method are: embryos at 15–30 min post oviposition, two incubation steps in 100% deuterated methanol at − 7 °C and − 14.5 °C, and rapid cooling. Eggs are recovered by rapid warming with concomitant dilution of cryoprotectant. Eggs of genetically modified A. stephensi and of A. gambiae were also successfully cryopreserved. This enabling methodology will allow long-term conservation of mosquitoes as well as acceleration of genetic studies and facilitation of mass storage of anopheline mosquitoes for release programs.

PeerJ ◽  
2020 ◽  
Vol 8 ◽  
pp. e10174
Author(s):  
Hitomi Takahashi ◽  
Yutaka Kodama

Genetic modifications such as mutation and transformation are powerful tools to study the function of genes and proteins in the model liverwort Marchantia polymorpha, but maintaining the resulting germplasm requires a practical, reliable method. Cryopreservation methods allow researchers to maintain mutant and transgenic lines of M. polymorpha. To date, two methods have been developed for cryopreservation of M. polymorpha gemmae: in the first method, unencapsulated gemmae are stored in liquid nitrogen at −­196 °C, and in the second method, encapsulated gemmae are stored in liquid nitrogen at −­196 °C or a deep freezer at −80 °C. In the present study, we developed a simple method named CRUNC (cr yopreservation of un en c apsulated gemmae), which can be used to store unencapsulated, dried gemmae of wild-type and transgenic M. polymorpha lines in liquid nitrogen and in freezers at −80 °C and −20 °C. Using the CRUNC method, we observed a high recovery rate (as high as 100%) and successful long-term (5 months) storage of the gemmae. Therefore, the CRUNC method is practical for maintaining valuable M. polymorpha germplasm.


2020 ◽  
Vol 287 (1931) ◽  
pp. 20201093 ◽  
Author(s):  
K. L. Miazgowicz ◽  
M. S. Shocket ◽  
S. J. Ryan ◽  
O. C. Villena ◽  
R. J. Hall ◽  
...  

Models predicting disease transmission are vital tools for long-term planning of malaria reduction efforts, particularly for mitigating impacts of climate change. We compared temperature-dependent malaria transmission models when mosquito life-history traits were estimated from a truncated portion of the lifespan (a common practice) versus traits measured across the full lifespan. We conducted an experiment on adult female Anopheles stephensi , the Asian urban malaria mosquito, to generate daily per capita values for mortality, egg production and biting rate at six constant temperatures. Both temperature and age significantly affected trait values. Further, we found quantitative and qualitative differences between temperature–trait relationships estimated from truncated data versus observed lifetime values. Incorporating these temperature–trait relationships into an expression governing the thermal suitability of transmission, relative R 0 ( T ), resulted in minor differences in the breadth of suitable temperatures for Plasmodium falciparum transmission between the two models constructed from only An. stephensi trait data. However, we found a substantial increase in thermal niche breadth compared with a previously published model consisting of trait data from multiple Anopheles mosquito species. Overall, this work highlights the importance of considering how mosquito trait values vary with mosquito age and mosquito species when generating temperature-based suitability predictions of transmission.


2012 ◽  
Vol 56 (5) ◽  
pp. 2546-2552 ◽  
Author(s):  
Long Cui ◽  
Zenglei Wang ◽  
Hongying Jiang ◽  
Daniel Parker ◽  
Haiyan Wang ◽  
...  

ABSTRACTThe recent emergence of artemisinin (ART) resistance inPlasmodium falciparumin western Cambodia, manifested as delayed parasite clearance, is a big threat to the long-term efficacy of this family of antimalarial drugs. Among the multiple candidate genes associated with ART resistance inP. falciparum, the sarcoplasmic/endoplasmic reticulum Ca2+-ATPase PfATP6 has been postulated as a specific target of ARTs. ThePfATP6gene harbors multiple single-nucleotide polymorphisms in field parasite populations, and S769N has been associated with decreased sensitivity to artemether in parasite populations from French Guiana. In this study, we used an allelic exchange strategy to engineer parasite lines carrying the S769N mutations inP. falciparumstrain 3D7 and evaluated whether introduction of this mutation modulated parasite sensitivity to ART derivatives. Using three transgenic lines carrying the 769N mutation and two transgenic lines carrying the wild-type 769S as controls, we found that S769N did not affectPfATP6gene expression. We compared the sensitivities of these parasite lines to three ART derivatives, artemether, artesunate, and dihydroartemisinin, in 18 biological experiments and detected no significant effect of the S769N mutation on parasite response to these ART derivatives. This study provides further evidence for the lack of association of PfATP6 with ART resistance.


2016 ◽  
Vol 2016 (14) ◽  
pp. 830-854
Author(s):  
Tzahi Y Cath ◽  
Ryan W Holloway ◽  
Leslie Miller-Robbie ◽  
Mehul Patel ◽  
Jennifer R Stokes ◽  
...  

2019 ◽  
Vol 43 (6) ◽  
pp. 347-354 ◽  
Author(s):  
Daniela Popp ◽  
Romanus Diekmann ◽  
Lutz Binder ◽  
Abdul R. Asif ◽  
Sara Y. Nussbeck

Abstract Various information technology (IT) infrastructures for biobanking, networks of biobanks and biomaterial management are described in the literature. As pre-analytical variables play a major role in the downstream interpretation of clinical as well as research results, their documentation is essential. A description for mainly automated documentation of the complete life-cycle of each biospecimen is lacking so far. Here, the example taken is from the University Medical Center Göttingen (UMG), where the workflow of liquid biomaterials is standardized between the central laboratory and the central biobank. The workflow of liquid biomaterials from sample withdrawal to long-term storage in a biobank was analyzed. Essential data such as time and temperature for processing and freezing can be automatically collected. The proposed solution involves only one major interface between the main IT systems of the laboratory and the biobank. It is key to talk to all the involved stakeholders to ensure a functional and accepted solution. Although IT components differ widely between clinics, the proposed way of documenting the complete life-cycle of each biospecimen can be transferred to other university medical centers. The complete documentation of the life-cycle of each biospecimen ensures a good interpretability of downstream routine as well as research results.


Genes ◽  
2021 ◽  
Vol 12 (1) ◽  
pp. 119
Author(s):  
Chioma Oringanje ◽  
Lillian R. Delacruz ◽  
Yunan Han ◽  
Shirley Luckhart ◽  
Michael A. Riehle

Mitochondrial integrity and homeostasis in the midgut are key factors controlling mosquito fitness and anti-pathogen resistance. Targeting genes that regulate mitochondrial dynamics represents a potential strategy for limiting mosquito-borne diseases. AMP-activated protein kinase (AMPK) is a key cellular energy sensor found in nearly all eukaryotic cells. When activated, AMPK inhibits anabolic pathways that consume ATP and activates catabolic processes that synthesize ATP. In this study, we overexpressed a truncated and constitutively active α-subunit of AMPK under the control of the midgut-specific carboxypeptidase promotor in the midgut of female Anopheles stephensi. As expected, AMPK overexpression in homozygous transgenic mosquitoes was associated with changes in nutrient storage and metabolism, decreasing glycogen levels at 24 h post-blood feeding when transgene expression was maximal, and concurrently increasing circulating trehalose at the same time point. When transgenic lines were challenged with Plasmodium falciparum, we observed a significant decrease in the prevalence and intensity of infection relative to wild type controls. Surprisingly, we did not observe a significant difference in the survival of adult mosquitoes fed either sugar only or both sugar and bloodmeals throughout adult life. This may be due to the limited period that the transgene was activated before homeostasis was restored. However, we did observe a significant decrease in egg production, suggesting that manipulation of AMPK activity in the mosquito midgut resulted in the re-allocation of resources away from egg production. In summary, this work identifies midgut AMPK activity as an important regulator of metabolism, reproduction, and innate immunity in An. stephensi, a highly invasive and important malaria vector species.


2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Yifat Ofir-Birin ◽  
Hila Ben Ami Pilo ◽  
Abel Cruz Camacho ◽  
Ariel Rudik ◽  
Anna Rivkin ◽  
...  

AbstractPathogens are thought to use host molecular cues to control when to initiate life-cycle transitions, but these signals are mostly unknown, particularly for the parasitic disease malaria caused by Plasmodium falciparum. The chemokine CXCL10 is present at high levels in fatal cases of cerebral malaria patients, but is reduced in patients who survive and do not have complications. Here we show a Pf ‘decision-sensing-system’ controlled by CXCL10 concentration. High CXCL10 expression prompts P. falciparum to initiate a survival strategy via growth acceleration. Remarkably, P. falciparum inhibits CXCL10 synthesis in monocytes by disrupting the association of host ribosomes with CXCL10 transcripts. The underlying inhibition cascade involves RNA cargo delivery into monocytes that triggers RIG-I, which leads to HUR1 binding to an AU-rich domain of the CXCL10 3’UTR. These data indicate that when the parasite can no longer keep CXCL10 at low levels, it can exploit the chemokine as a cue to shift tactics and escape.


Materials ◽  
2021 ◽  
Vol 14 (13) ◽  
pp. 3549
Author(s):  
Tulane Rodrigues da Silva ◽  
Afonso Rangel Garcez de Azevedo ◽  
Daiane Cecchin ◽  
Markssuel Teixeira Marvila ◽  
Mugahed Amran ◽  
...  

The urbanization process contributes to the growth of solid waste generation and causes an increase in environmental impacts and failures in the management of solid waste. The number of dumps is a concern due to the limited implementation and safe disposal of this waste. The interest in sustainable techniques has been growing in relation to waste management, which is largely absorbed by the civil construction sector. This work aimed to review plastic waste, especially polyethylene terephthalate (PET), that can be incorporated with construction materials, such as concrete, mortars, asphalt mixtures, and paving. The use of life-cycle assessment (LCA) is related, as a tool that allows the sustainability of products and processes to be enhanced in the long term. After analyzing the recent literature, it was identified that studies related to plastic wastes in construction materials concentrate sustainability around the alternative destination of waste. Since the plastic waste from different production chains are obtained, it was possible to affirm the need for a broader assessment, such as the LCA, providing greater quantification of data making the alternative processes and products more sustainable. The study contributes to enhance sustainability in alternative building materials through LCA.


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