Célia Seassau
email: celia.seassau@toulouse.inra.fr

Grégory Dechamp-Guillaume

Emmanuelle Mestries

Philippe Debaeke

Effect of nitrogen and water management on sunflower premature ripening caused by Phoma macdonaldii

Understand how cropping management system influences sunflower premature ripening to better explain the erratic occurrence of the disease in S.W France

Main headways

➢ Sunflower premature ripening syndrome (PR) is primarily caused by P. macdonaldii collar infection ➢ The disease severity is mainly influenced by crop management ➢ Cultivar susceptibility to the pathogen, high N-fertilization and rainfed management all favor P. macdonaldii infection and enhance PR

-Scientific context

➢ Collar girdling canker caused by P. macdonaldii is the primary cause of PR ➢ Sunflower premature death is characterized by sudden plant wilting during mid-to late summer followed by plant death a few weeks before normal maturity ➢ Collar infection at flower bud stage is the best way to reproduce PR plants in the field ➢ Recent investigations pointed out an impact of Nfertilization and water shortage on the disease severity.

-Results

What is the effect of sunflower crop management on the severity of attacks and PR induced by P. macdonaldii ?

➢ Cultivar susceptibility was the main factor influencing PR plants, cv. Heliasol RM being more sensitive than cv. Melody ➢ High N-fertilization seriously affected disease progress on both cultivars ➢ Water stress amplified nitrogen effect and stimulated disease progress

-Conclusion

  , Midi-Pyrénées government and Promosol association who supported this study. Also thanks to INRA technical staff : D. Raffaillac, M. Labarrère and C. Quinquiry and other people who participated to disease scoring.

➢

  Appropriate cultural practices could reduce sunflower PR ➢ Promising cultivars should be screened at high N supplies and under water-limited conditions post flowering, in resistance tests during breeding programmes 2 -Methods (field study) at the plant collar Cotton and aluminum avoid the disc desiccation 9 th International Plant Disease Conference, Tours, France, December 2009


PhD Marije Nouwen
email: marije.nouwen@kuleuven.bemathildeduflos

PhD Mathilde Duflos
email: mathilde.duflos@univ-tours.fr

Using the intergenerational solidarity framework to understand the grandparentgrandchild relationship: A scoping review

Keywords: TikTok, Grandparenting, Grandchildren, COVID-19, Intergenerational solidarity, Family display

published or not. The documents may come     Copyright Displaying intergenerational solidarity on TikTok during the COVID-19 pandemic: understanding the implications in the grandparent-grandchild relationship

Displaying intergenerational solidarity on TikTok during the COVID-19 pandemic:

Understanding the implications in the grandparent-grandchild relationship.

The habits of families are affected during the COVID-19 pandemic, with limitations to socialization or visits. Households are separated on a worldwide scale, due to barrier measures like keeping physical distance and self-isolation. Since the beginning of the pandemic, older persons have been identified as a group particularly exposed to health risks [START_REF] Settersten | Understanding the effects of Covid-19 through a life course lens[END_REF] especially those with pre-existing health conditions [START_REF] Ayalon | There is nothing new under the sun: ageism and intergenerational tension in the age of the COVID-19 outbreak[END_REF]. Sanitary measures were often associated with the imperative to protect them. Vervaecke and Meisner (2021) noted that the risks for older adults were often over-simplified, leading to an increase of paternalistic actions to protect older people from the virus. Younger generations were attributed an important role in keeping older generations safe. For families, COVID-19 health risks often resulted in different generations keeping physical distance which led to adaptations in the relationship pattern and in the enactment of intergenerational solidarity [START_REF] Gilligan | Multigenerational social support in the face of the COVID-19 pandemic[END_REF].

To avoid the transmission of the virus, grandchildren had limited face-to-face contact with their grandparents. Many families that lived in separate households turned to technology to maintain contact during the pandemic [START_REF] Gilligan | Multigenerational social support in the face of the COVID-19 pandemic[END_REF]. Also, older adults between 65 and 74 years old were significantly more likely to use technologies and social media to maintain their connections while being physically separated from their friends and family [START_REF] Cirilli | Elderly People, Covid-19 and Technologies: a Qualitative Research[END_REF]. Most grandparents stated the importance of using technologies during the pandemic to reduce feelings of loneliness, even though they prefer physical meetings [START_REF] Watson | Enacting intimacy and sociality at a distance in the COVID-19 crisis: the sociomaterialities of home-based communication technologies[END_REF]. The uptake and use of social media to maintain family connections emphasizes the role of such platforms for intergenerational solidarity between grandparents and grandchildren. Some platforms provide support for interpersonal communication, like sending text messages or video calling, while other platforms put on display how grandparents and grandchildren experience the pandemic. In this paper, we consider the latter and focus on TikTok, a social media platform that features short videos, mostly for entertainment. During the pandemic, the number of TikTok users (Statista, 2020a) and scholarly attention has increased. Moreover, media reported on TikTok as a platform for grandparents and grandchildren to stay connected [START_REF] Dodgson | TikTokers are dancing with their grandparents for content, and it could help curb the pandemic of loneliness among older people[END_REF][START_REF] Page | Grandparents are dancing with their grandkids on TikTok. People can't get enough[END_REF]. TikTok thus promises to display different ways in which intergenerational solidarity unfolds between grandparents and grandchildren. The Intergenerational Solidarity Framework, which is concerned with the social cohesion and belonging across generations [START_REF] Bengston | Intergenerational Solidarity: Strengthening Economic and Social Ties[END_REF], and the concept of 'family display' [START_REF] Finch | Displaying families[END_REF] theoretically inform our study. This paper presents a qualitative content analysis of the display of different dimensions of intergenerational solidarity between grandparents and grandchildren during the COVID-19 pandemic in 2020, on TikTok.

The intergenerational solidarity framework

The widespread use of social media during the last decades has mediated family relationships [START_REF] Cabalquinto | Home on the move: negotiating differential domesticity in family life at a distance[END_REF][START_REF] Carvalho | Family functioning and information and communication technologies: How do they relate? A literature review[END_REF], with the potential to enrich grandparent-grandchild relationships [START_REF] Bangerter | Turning points in long distance grandparentgrand-child relationships[END_REF]. Mobile social media enhanced family solidarity among family members who live in separate households [START_REF] Hänninen | Refamilisation in the broadband societythe effects of ICTs on family solidarity in Finland[END_REF]. Frequent mobile phone calls and text messages increased the feeling of clo-seness while members were physically separated [START_REF] Christensen | Connected presence" in distributed family life[END_REF], also during the pandemic (McDarby et al., 2021). In sum, social media play an important role in maintaining feelings of belonging in families and displaying intergenerational solidarity.

The Intergenerational Solidarity Framework (ISF; [START_REF] Bengtson | Intergenerational Solidarity in Aging Families: An Example of Formal Theory Construction[END_REF] stipulates six dimensions of intergenerational solidarity (i.e., affectual solidarity, associational solidarity, consensual solidarity, functional solidarity, normative solidarity, structural solidarity) that are presented in Table 1. These six dimensions stem from interpersonal relationship theories [START_REF] Heider | The Psychology of Interpersonal Relations[END_REF] and from studies in sociology on normative group commitment and group solidarity [START_REF] Tönnies | Community and Society[END_REF]. The studies with the ISF have gradually moved their interest from functional dependence, from the oldest generation on younger generations, to a focus on intimacy as a cohesive force in families [START_REF] Hammarström | The construct of intergenerational solidarity in a lineage perspective: A discussion on underlying theoretical assumptions[END_REF] sometimes emphasizing how emotional and social support between generations extends to the use of communication technology [START_REF] Azevedo | Intergenerational solidarity or intergenerational gap? How elderly people experience ICT within their family context[END_REF].

Table 1. Six components of the Intergenerational Solidarity Framework [START_REF] Bengtson | Intergenerational Solidarity in Aging Families: An Example of Formal Theory Construction[END_REF].

Component Definition Examples

Affectual solidarity

Feelings of closeness and reciprocity in emotions between members of a family Saying "I love you", hugging or making jokes

Associational solidarity

Different types and patterns of contact and interactions within a family

Visiting each other, dancing together or sharing a meal

Consensual solidarity

Normative solidarity

Expectations toward filial and parental duties, and to the sense of obligation related to a person's role within the family and compliance with family norms and values

Respecting filial piety and family values

Structural solidarity

Opportunities for intergenerational contact facilitated by geographical proximity Living in the same house, travelling to visit each other or general health

The ISF proposes that different feelings and behaviors between two generations, including grandparents and grandchildren, define their relationship [START_REF] Silverstein | Intergenerational solidarity and the grandparent role[END_REF] among various cultures and at all ages [START_REF] Duflos | Using the intergenerational solidarity framework to understand the grandparent -grandchild relationship : a scoping review[END_REF]. Moreover, Gilligan et al.

(2020) argue for the relevance of using this model to investigate grandparenting during the pandemic as it provides a reading grid to understand the effect of the pandemic on intergenerational cohesion. Although positive connotations are often implied with regards to intergenerational solidarity, ambiguity [START_REF] Hammarström | The construct of intergenerational solidarity in a lineage perspective: A discussion on underlying theoretical assumptions[END_REF] or ambivalence [START_REF] Lüscher | Intergenerational ambivalence: Beyond solidarity and conflict[END_REF]) also form part of intergenerational relationships. Related to technology use, [START_REF] Ivan | Family conflicts and technology use: The voices of grandmothers[END_REF] reported on grandmothers that expressed concern about their grandchildren's exposure to potentially harmful online content as well as pressure to use new platforms. Also, grandparents express concerns about regular but shallow contact with their grandchildren via chat applications or feelings of exclusion [START_REF] Azevedo | Intergenerational solidarity or intergenerational gap? How elderly people experience ICT within their family context[END_REF]. The

The use of information and communication technology (ICT) and social network services (SNS) to interact between generations has previously been studied under the associational solidarity dimension only [START_REF] Peng | Use of communication technology to maintain intergenerational contact: toward an understanding of 'digital solidarity[END_REF], or specifically with geographically separated extended families in the form of written chat messages on platforms like WhatsApp or Facebook [START_REF] Taipale | Intergenerational solidarity[END_REF]. Grandparents have been found to appreciate platforms like Skype when their grandchildren moved away [START_REF] Taipale | Intergenerational solidarity and ICT usage: Empirical insights from Finnish and slovenian families[END_REF]. Sharing affectual solidarity was only possible when grandparents and grandchildren used the same platforms [START_REF] Taipale | Intergenerational solidarity[END_REF] and agreed on appropriate social conduct (e.g. refraining from embarrassing comments on SNS) [START_REF] Taipale | Intergenerational solidarity and ICT usage: Empirical insights from Finnish and slovenian families[END_REF]. Functional solidarity was reported to be more complex when grandparents and grandchildren where in geographically separate places, compared to physical meetings [START_REF] Taipale | Intergenerational solidarity[END_REF]. Structural solidarity between grandparents and grandchildren over digital devices was reported to become more complex due to physical deterioration (e.g. poor vision or finger control) [START_REF] Taipale | Intergenerational solidarity and ICT usage: Empirical insights from Finnish and slovenian families[END_REF]. Studies that investigated intergenerational solidarity between grandparents and grandchildren using ICT and SNS are scarce. However, there are indications that each dimension of intergenerational solidarity can be observed on SNS. For instance, expressing love (i.e. affectual solidarity) can occur during a visit as well as in an Instagram post. Considering that many family interactions moved online during the pandemic, there is a greater urgency to study intergenerational solidarity on SNS. Although most studies on the grandparent-grandchild relationship under the ISF were quantitative studies [START_REF] Duflos | Using the intergenerational solidarity framework to understand the grandparent -grandchild relationship : a scoping review[END_REF], this paper presents a qualitative content analysis to understand how the different dimensions unfold on TikTok during the COVID-19 pandemic.

Displaying family on TikTok

The social media platform TikTok ("Dou Yin" in China) was launched internationally in 2017 and allows for creative video making [START_REF] Bresnick | Intensified Play: Cinematic study of TikTok mobile app[END_REF]. Like other social media, the content production on TikTok can be viewed as an iterative process between the physical world that people inhabit and the conversations that happen online (Alder, 2020). TikTok features videos are different from those shared on other video platforms, mainly in size and popularity [START_REF] Chen | A study on the characteristics of douyin short videos and implications for edge caching[END_REF]. Whereas Instagram or Facebook users consume content of people or organizations that they follow, TikTok users land immediately on the 'For You'page where videos are presented based on the platform's algorithms that estimate which videos are most likely to retain the user's attention [START_REF] Schellewald | Communicative Forms on TikTok: Perspectives From Digital Ethnography[END_REF]. The platform features of TikTok allow users to imitate and replicate existing content [START_REF] Zulli | Extending the Internet meme: Conceptualizing technological mimesis and imitation publics on the TikTok platform[END_REF]) like sounds, movements and challenges (e.g. dance, fitness or magic trick challenges) which users often personalize [START_REF] Klug | It took me almost 30 minutes to practice this[END_REF]. Prior research on TikTok users has mainly focused on youth, who compose the largest user group of the platform (Statista, 2020b). TikTok influences younger users' social and cultural lives. Pre-adolescents are more motivated to use TikTok to socialize than adolescents, as adolescents mainly pursue fame, self-expression and social recognition on TikTok [START_REF] Bucknell Bossen | Uses and gratifications sought by pre-adolescent and adolescent TikTok consumers[END_REF]. During the pandemic, when people were invited to stay home to comply with lockdown policies, teenage bedrooms [START_REF] Kennedy | If the rise of the TikTok dance and e-girl aesthetic has taught us anything, it's that teenage girls rule the internet right now': TikTok celebrity, girls and the Coronavirus crisis[END_REF] and other private spaces were often portrayed publicly.

The blurring boundaries between private and public display of private space is equally important for the grandparent-grandchild relationship on TikTok. In private conversations, over Skype for instance, the possibility to see family members contributed to a meaningful relationship between grandchildren and grandparents during the pandemic [START_REF] Watson | Enacting intimacy and sociality at a distance in the COVID-19 crisis: the sociomaterialities of home-based communication technologies[END_REF]. The need to 'display family' [START_REF] Finch | Displaying families[END_REF] to others increases when family practices change, like the impossibility to celebrate birthdays in each other's presence. The desired audience can be both internal and external to the family. Visual display of grandparentgrandchild relationships has been studied on social media platforms, like Instagram and Facebook [START_REF] Barnwell | Captured and captioned: Representing family life on Instagram[END_REF][START_REF] Shannon | #Family: Exploring the display offamily and family leisure on Facebook and Instagram[END_REF], chat applications (Cabal-quinto, 2020) or Skype [START_REF] Share | Displaying and performing: Polish transnational families in Ireland skyping grandparents in Poland[END_REF]. SNS expand the target audience of family relationships on display from family members to peers, and even strangers. The public aspect of TikTok possibly influences what grandparents and grandchildren share about their relationship, with the (unconscious) intention to conform with normative family representations [START_REF] Barnwell | Captured and captioned: Representing family life on Instagram[END_REF][START_REF] Shannon | #Family: Exploring the display offamily and family leisure on Facebook and Instagram[END_REF].

Our study builds on previous work concerned with grandparents and grandchildren on SNS. We empirically scrutinize the mainstream user group (i.e. youth) of TikTok while adding another generation to the empirical scopein their respective roles as grandchildren and grandparents. Studying two generations on TikTok advances previous work in two ways.

On the one hand, our work aims to empirically understand the growing but relatively unknown user group of older adults, in their role of grandparent, on the platform. Adults over 50 years old represented 11% of total users on TikTok, a number that has increased since March 2020 (Statista, 2020b). We ground our decision to investigate the grandparentgrandchild relationship, specifically, based on previous work that suggests that in interpersonal contacts between two generations, individual preferences and practices are negotiated towards the habits of the other [START_REF] Fernández-Ardèvol | I do it my way": Idioms of practice and digital media ideologies of adolescents and older adults[END_REF]. Also, younger generations have been found to empathize with retirement-aged SNS users through hypothetical grandparent-grandchild relationships [START_REF] Harley | YouTube and intergenerational communication: The case of Geriatric1927[END_REF].

On the other hand, our study broadens the empirical attention on young people from peer interactions to family relationships. The research goals expand the concept of 'displaying family' [START_REF] Finch | Displaying families[END_REF] on TikTok where families simultaneously maintain feelings of belonging to each other, and a space where they can be recognized by others as family members (Merla & Papanikolaou, 2021). The ISF [START_REF] Bengtson | Intergenerational Solidarity in Aging Families: An Example of Formal Theory Construction[END_REF] endorses the goal of our study as it equally highlights the efforts of grandparents and grandchildren to maintain their relationship.

Our research paper is guided by the following research question: How do grandparents and grandchildren display their relationship on TikTok during the COVID-19 pandemic?

Considering the restrictions that this pandemic entails, we will focus equally on the use of materials and physical arrangements used by grandparents and grandchildren to enforce barrier measures when they are together.

Methods

To understand how grandparents and grandchildren display intergenerational solidarity, we conducted a qualitative media analysis [START_REF] Altheide | Qualitative Media Analysis[END_REF] of TikTok videos. Individual users were not informed about their selection for our study as this would entail a disproportionate distribution of information for publicly accessible video content. We took measures to protect the privacy of participants by excluding usernames and personal identifiable information in the data file. All videos were deleted from the researchers' personal devices after analysis and stored on a network drive managed by the first author's university. Prior to the study, the research protocol received approval from the social and societal ethics committee at the faculty of the first author.

Data collection

To search and download videos, we created a research account in January 2021 to avoid influencing TikTok's algorithms based on previous search queries [START_REF] Zulli | Extending the Internet meme: Conceptualizing technological mimesis and imitation publics on the TikTok platform[END_REF].

To compound the data corpus, we followed a common practice in online research by combining hashtags for data selection [START_REF] Jackson | Hijacking #MYNYPD: Social Media Dissent and Networked Counterpublics[END_REF]. We identified and combined five relevant hashtags (#grandchild, #grandparent, followed by #quaran-tine, #lockdown, or #corona) to answer the research question. Selected videos were pub-lished when the pandemic hit globally, between March and December 2020. We only included videos in languages we mastered (i.e. Dutch, English, French, German, Spanish). The criterion for including videos in the data corpus was that at least one grandchild or grandparent should be visible or, that users referred to their role as grand-parent or grandchild. For each search query, we estimated to select around one hour of video material (approximately up to 70 videos). The selection process is described in Figure 1. We first downloaded the videos from TikTok. Then, we added general information about the video in the data corpus (i.e. serial number, search query letter code, search date, upload date, amount of comments, amount of likes, amount of shares, hashtags, caption, duration in seconds, URL).

Data analysis

We transcribed all videos, with descriptions of the different components of the video (e.g. music, spoken words, written text, filter), and included analytic memos to textually interpret the moving images [START_REF] Saldaña | The coding manual for qualitative researchers[END_REF]. The transcripts and memos amounted to approximately 71,000 words or 142 A4-pages. Then, we proceeded with three coding rounds of the six dimensions of the ISF, following an adaptive theory approach that builds on principles of grounded theory while relying on prior theory to order the analysis [START_REF] Layder | Sociological Practice. Linking Theory and Social Research[END_REF]. The first coding round unfolded over two stages. In the first stage, we coded the videos of one search query individually. Then, we checked for differences in interpretations in thorough discussions, which resulted in a refined set of concrete coding guidelines for the second stage. We then divided the coding dimensions for the remaining videos and iteratively engaged in discussions when doubts about interpretations arose. At the end of the first coding round, each video had a main code that related to one of the six dimensions of the ISF that structured the subsequent two coding rounds. In the second coding round, we looked for subthemes for each of the six dimen-sions inductively. Author two coded affectual, consensual, and structural solidarity, and author one coded associational, functional, and normative solidarity. Both authors reg-ularly discussed the relations between and categorizations of the different sub themes. In the third coding round, both authors revisited the sub themes in the assigned dimensions to look for patterns in and between the different sub themes and dimensions. The final coding book can be found in Table 2. In a final step, the list of codes was reviewed by means of upload date, number of comments, likes and shares, duration of the video, use of music, use of filters, identified cultural background and the person sharing the video. The goal of this final step was to explore the features of the platform, and its role in the performance of the grandparentgrandchild relationship on TikTok. Table 3 summarizes the description of the data corpus.

Table 3. Description of the videos related to TikTok features (left column) and summary of user characteristics (right column).

TikTok platform Users

Duration

Affectual Solidarity

Affectual solidarity was the most present dimension (n = 128). 30 videos coded under the subtheme "affectionate gestures", mostly shared by parents (n = 15), showed grandparents and grandchildren cuddling, hugging, and kissing each other. Affectionate gestures were specifically adapted to respect barrier measures in six videos of grandparents or grandchildren dressed up in body suits (Figure 2) and in seven videos where they used a plastic wall to protect each other (Figure 3). Subscripts accompanying these videos highlighted the longevity of the time they were not able to hug (e.g. 'finally' or 'haven't hugged my grandparents in five months <3'), suggesting that hugging is an important gesture for their relationship. 30 videos were about "celebration and gifts", as families celebrated birthdays (n = 18), Christmas (n = 4), Easter (n = 2), grandmother's day (n = 2), or wedding or wedding anniversaries (n = 2). In most ofthe videos (n = 23), they made adaptations to respect physical distance, by remaining outside or celebrating over video calls.

16 videos showed "fun and humor" in the grandparent-grandchild relationship, like when a granddaughter told her grandparents that they would be "viral" on TikTok, and they answered that they refused to be "corona viral". Moreover, other videos displayed pranks, excerpts from video calls where families displayed their struggles with technology or did antics to make a baby laugh. This subtheme also includes the video that received the most comments (n = 89,300), likes (n = 6,800,000) and shares (n = 501,900) in the corpus. This video featured a grandmother who yelled at her husband because he supposedly harassed his granddaughter. However, the granddaughter indicated in the written text on the video that she instigated it all.

The subtheme "art and cards" (n = 6) presents grandparents or grandchildren creating art pieces for each other or writing cards to their grandparents in order to continue to express their feelings for each other when they were unable to be together.

Another aspect of affectual solidarity was expressed through "grief" as an adult granddaughter filmed herself crying after her grandparents died of COVID-19. The subtheme "memories" (n = 30) puts forward the importance of family pictures. For instance, when grandchildren re-enacting pictures from their childhood with their grandparents (n = 4) and compilations of pictures or videos (n = 25) with subscripts that expresses affection or missing out (e.g., "I can't take the pain of not seeing my grandson"). In the subtheme "missing each other) (n = 5), we see what grandparents and grandchildren do when they miss each other, like a toddler trying to open the door to go to the grandparents' house or a grandfather using a mirror to reflect the sun and send signals to his grandson. In other videos, grandchildren filmed themselves having strong emotions while talking to the camera or using text to accompany their emotions; e.g.:

I just realized that every single day that passes, it's one day closer to me losing my grandparents. I feel that their time might be coming. I'm not ready to lose them but eventually that day will come, and I'm going to have to say goodbye to the people who helped shape me into the person I am today…

Associational Solidarity

Associational solidarity was the second most present dimension (n = 91), with eight subthemes that explain the type and frequency of contact between grandparents and grandchildren during the COVID-19 pandemic. The first subtheme was "TikTok challenges"

(n = 34) and displayed grandparents and grandchildren participating in these challenges. Most of these videos were uploaded by grandchildren (25 of 34). The challenge with the highest number of likes in our data set included the song Laxed (siren beat) by Jawsh 685 (n = 7), an instrumental song that is featured in 1.6 million TikTok videos. The choreography is slow and mainly involves upper body movements, which made the challenge accessible to grandparents seated in a wheeled walker, or in a wheelchair (see Figure 4). Six of the seven videos were danced outside and were posted between the end of April 2020 and the beginning of July 2020. Only two videos coded under 'TikTok challenge' included a challenge without dancing (i.e. culinary challenge, game challenge). In the subscript of challenge videos, grandchildren expressed their love for their grandparents or motivated TikTok users to like their videos. The grandchildren also addressed other TikTok users with affectual messages, to display warm feelings towards their grandparents.

The second subtheme under associational solidarity was "keeping entertained" (n =

33)

and included videos that showed how grandparents and grandchildren spent time together during quarantine. 15 videos were shared by grandchildren, six by parents, ten by grandparents and two by unknown persons. The most common activity involved dancing. Ten videos displayed children playing, dancing or singing. Four videos did not include grandchildren but featured grandparents dancing together. These videos were usually uploaded by a grandchild but often recorded by the grandparents themselves. The captions refer to how grandparents kept entertained during quarantine for instance "dancing through quarantine" or "what happens when grandparents get quarantined!" Videos that stand out showed how grandparents took on rather extreme measures to entertain their grandchildren, like shooting a nerf gun at their grandchild.

Door visits (n = 9) and garden visits (n = 7) are two other sub-themes of associational solidarity that show how grandparents and grandchildren visit each other without entering the house, for instance by standing at the front door, with a window in between, or in a garden.

The content of the videos often refers to grandchildren missing their grandparents. During garden visits, food and beverages were often shared, for instance with grandparents sitting inside and grandchildren sitting outside. Two videos, uploaded by the parents, showed toddlers and focused on greeting each other.

The remaining subthemes include "beauty salon" (n = 4) with videos of doing each other's hair and make-up, "phone and video calls" (n = 3) with videos that highlight the act of calling over phone or video, "baking together" with two videos where grandchildren and grandparents bake a cake or bread together, and a final video coded under "accidental meeting" where a granddaughter and grandparents bump into each other at the supermarket.

Consensual Solidarity

Consensual solidarity was the least present dimension (n = 3). These videos were shared by adolescent or adult grandchildren. Two videos were about differences in political views. One video showed how a granddaughter cut her grandfather's Trump Card in pieces before handing it back to him. This video received the second most comments in our data corpus. Another video showed a granddaughter inciting a grandmother to dance with her on a song with lyrics that promoted the use of marijuana. When the grandmother heard the lyrics, she stopped dancing and walked away. In the last video, a grandson filmed himself while coming out to his grandmother by phone and she expressed him her unconditional love:

It makes absolutely no difference to me. You are my grandson, in spite of everything.

I love you as you are and no matter what… I will never change the way I feel about you. I'm okay with it. Just be yourself! Just don't worry about anything.

Functional Solidarity

Functional solidarity was identified in 49 videos. Most of these videos included grandchildren or grandparents who provided instrumental support to each other (n = 25). In Another subtheme under functional solidarity is related to food. When grandchildren brought food, that was usually homemade, to their grandparents (n = 9), videos were about adolescent or emerging adult grandchildren. In the videos about grandparents who prepared and brought food to their grandchildren (n = 7), three were about adolescent grandchildren who, in turn, expressed love towards their grandparents. The food was handed over in the car or at the doorstep. Three videos, uploaded by grandparents, showed them feeding a baby grandchild. Another video, uploaded by a parent, includes a grandmother who brings homemade cookies to her toddler grandchild.

The last subtheme of functional solidarity includes gifting or bringing by non-essential items (n = 9; all uploaded by grandchildren). In eight videos, grandchildren offered different items to their grandparents (e.g., hand sanitizer, sweets, cards, clothes). The reactions of grandparents ranged from being ecstatic and grateful to bursting into tears and being disappointed that the grandchild could not come inside. Most of the subscripts for the videos coded under functional solidarity emphasized that this type of intergenerational solidarity required little effort from the giver, while having a big impact on the well-being of the receiver.

Normative Solidarity

Normative solidarity was identified in 38 videos, all of which put forward different expectations about how grandparents and grandchildren should behave towards each other.

Most videos were themed around meeting baby grandchildren for the first time (n = 21), hinting towards the importance that new grandparents and parents give to the addition of a new generation to the family. 11 of these videos were uploaded by parents and six were uploaded by grandparents. In one of these videos, a grandmother emphasized that she was tested negative to COVID-19 before holding her grandchild. Similarly, other videos that showed grandparents meeting their grandchildren inside (n = 6) emphasized that grandparents had to wait for a long time and grandparents portrayed strong emotional reactions, like crying when holding the baby. Other videos showed how grandparents met their grandchild through windows (n = 5), in outdoor spaces (n = 6), or at the airport. Two videos stressed that the baby was named after the grandfather or great grandfather. One video showed that the grandmother prepared clothing for her newborn grandchild. Not all videos emphasized that grandparents waited or altered this moment due to COVID-measures, although many captions stated that grandparents were only able to visit a newborn after some months (e.g., "After being quarantined for months, Nana & Papa get to meet their granddaughter for the first time." To some grandparents, that situation was heartbreaking and it seemed that holding grandchildren was more important than the possibility to see them (e.g., "Please stay home during quarantine. I missed the birth of my only grandchild. I've never held her & have to stay 6' ft back to see her"). Like the subtheme "affectionate gestures" under affectual solidarity, the longevity and importance of physical touch becomes explicit

In addition to meeting baby grandchildren for the first time, five videos included pregnancy announcements to the soon-to-be (great-)grandparents (n = 5). Pregnancy announcements happened during a video call, at the front door, in the living room or by posting a series of pictures (e.g., the echo, the parents). The announcement was usually accompanied by a material token, like a mug that said "Just when grandparents think their work is finished someone calls them great" or a positive pregnancy test.

Normative solidarity was also reflected by grandchildren stating that they were the "favorite grandchild" (n = 6) when they received or expected special treatments. In three videos, grandchildren played with the audio to emphasize that they were their grandparents' favorite grandchild. Other videos showed how grandparents' expectations did not match the grandchildren's needs, for instance when grandparents did not appreciate the special gestures of their grandchildren or when a grandchild received less attention than other grandchildren.

All videos hinted towards how being a grandchild or grandparent is often performed in relation to other grandchildren, rather than a simple one-on-one relationship.

The last subtheme under normative solidarity includes videos where grandchildren paid special attention to protect their grandparents from getting infected by the COVID-19 virus (n = 4). All videos featured and were uploaded by adolescents. These videos showed a granddaughter running after her grandfather to check if he was carrying his face mask and hand sanitizer before driving off, grandparents trying out thermometers, a celebrity convincing TikTokers respect the measures in order to protect all grandparents, and a grandchild who wear a gas mask when visiting her grandmother inside. These examples show additional barrier measures that could be observed in other videos like wearing body suits (see affectual solidarity) or keeping physical distance.

Structural Solidarity

Structural solidarity was highlighted in 20 videos. These videos were mostly shared by grandchildren (75%), followed by parents (15%), and only one of them was shared by a grandparent. Another video was shared by the partner of a grandchild who filmed the grandparent-grandchild interaction. Structural solidarity was expressed through focusing on the health of a grandparent and how families adapted their interactions (n = 10), on traveling to visit each other (n = 7), like this granddaughter who commented her video: "So I haven't seen my grandparents in months due to the coronavirus. So now that Boris relaxed the rules, we traveled an hour and a half to see them… They had no idea! They were so excited!"

Other videos of this dimension focused on the living arrangement, either when grandchildren had to move from a shared household to protect their grandparents (n = 1) or when grandchildren spent the lockdown with their grandparents to take care of them (n = 2) by, for instance, keeping them company: "My grandma told me mom I'm her 'ray of sunshine' during this dark time. My heart is broken, make them viral". This comment combines the grandchild's emotional involvement during lockdown with a call for other TikTok users to comment, like, or share the video.

COVID-19 protective measures

Among the 328 videos of the present corpus, 72 could not be associated with adaptations related to COVID-19 due to their specificity (e.g., slideshow of memories prior to the pandemic, phone call), 127 videos showed grandparent-grandchild interactions without any kind of protective measures and the last 129 videos showed grandparent and/or grandchildren who used one or more protections to limit the transmission of the virus. Barrier

Discussion

This qualitative content analysis aimed to study the display of the different dimensions of intergenerational solidarity among grandparents and grandchildren on TikTok during the COVID-19 pandemic. Although most of the videos (over 60%) were posted by grandchildren, our analysis also yields insights into grandparents' use of TikTok (about 20% of the videos). The analysis identified videos for each dimension of the ISF, with some dimensions more populated by grandparent-shared content than others. In this discussion, we frame our findings in previous work on ISF as well as the implications for family display in the context of the pandemic. During this period, families were challenged to renegotiate their relationship and responsibilities towards each other, and family norms and values became scrutinized [START_REF] Ellerich-Groppe | Young for old-Old for young? Ethical per-spectives on intergenerational solidarity and responsibility in public discourses on COVID-19[END_REF]. Moreover, family relationships tend to become more openended in light of such events, which intensifies the need to display relationships that matter [START_REF] Finch | Displaying families[END_REF], like the grandparent-grandchild relationship.

Spending longer periods without touching each other is a common concern and frustration expressed in our data corpus, in line with the difficulties expressed by transnational families in previous studies [START_REF] Cabalquinto | Home on the move: negotiating differential domesticity in family life at a distance[END_REF]. Notwithstanding, some videos featured creative and COVID-compliant ways to maintain physical contact when the urgency to hug became increasingly unbearable after weeks and months in lockdown. These videos displayed hugging (i.e. affectual solidarity) as actions that confirm their sense of belonging as grandparents and grandchildren. Moreover, the urgency to hold newborns was also made explicit in videos displaying normative solidarity. In some videos, grandparents did not wait to hold their newborn grandchild even when infants were susceptible to be infected [START_REF] Mehta | SARS-CoV-2 (COVID-19): What Do We Know About Children? A Systematic Review[END_REF], especially in familial clusters [START_REF] Posfay-Barbe | COVID-19 in Children and the Dynamics of Infection in Families[END_REF]. In other videos, grandparents displayed their struggles with withdrawing from holding their newborn grandchild. Motivations to not postpone physical touch during the first meeting with a grandchild or to display it on TikTok might be the special circumstances in which the grandchild was born. When grandparents cannot engage themselves in activities they consider of value, it becomes especially meaningful to display 'grandparenting' during key life moments (Ducu, 2020).

Our findings revealed that families were having a good time together despite the lockdown, which is at the heart of grandparenting [START_REF] Neugarten | The Changing American Grandparent[END_REF]. This came forward in our data corpus in the dimensions of affectual and associational solidarity. Our results suggest that families celebrated special occasions in offline and online settings. Indeed, families were found to organize online celebrations during the pandemic, that would not have been organized online in pre-pandemic times [START_REF] Watson | Enacting intimacy and sociality at a distance in the COVID-19 crisis: the sociomaterialities of home-based communication technologies[END_REF]. Celebrating special occasions, then, is important to display family to other family members. Organizing celebrations to include as many people as possible confirms that the people present are part of the family [START_REF] Finch | Displaying families[END_REF], which becomes especially important when sanitary circumstances disrupt their festive habits. Also, the results of this study showed families' willingness to do dance challenges together. Associational solidarity was expanded by trends on TikTok, just like other social media have been found to enrich interactions between remote family members [START_REF] Taipale | Intergenerational solidarity and ICT usage: Empirical insights from Finnish and slovenian families[END_REF]. From our data it is not possible to compare the prevalence of dance challenges performed by grandparents and grandchildren with pre-COVID times.

However, the finding suggests that young TikTok users can appreciate intergenerational content and that this content is meaningful to all users of the platform [START_REF] Harley | YouTube and intergenerational communication: The case of Geriatric1927[END_REF].

Consensual solidarity was scarcely present in our analysis and suggests that TikTok is not a place to share different views or opinions explicitly in relation to the grandparentgrandchild relationship. Across the different dimensions, however, there were some indications about different conceptions of the health risks of grandparents mostly in line with the general discourse to protect elderly people from the virus (Meisner, 2021). In the associational solidarity dimension, this was most obvious when grandchildren refused to enter their grandparents' house, even when invited to come inside. Moreover, a small number of videos explicitly featured the efforts of grandchildren to protect their grandparents by different means under normative solidarity. Cooking and delivering meals to each other also seemed important and appeared as a privileged way to support each other during the pandemic. Both grandparents and grandchildren took part in this type of functional solidarity, while in the literature, grandparents, and especially grandmothers, are more likely to feed their grandchildren than the opposite [START_REF] Maijala | Grandparental love: a challenge or richness?[END_REF].

The results of our study pointed out that grandchildren were more likely than grandparents to share videos in which they took actions in protecting their grandparents, maybe because they need approval from their peers on SNS [START_REF] Cipolletta | How can adolescents benefit from the use of social networks? The iGeneration on Instagram[END_REF] or because they want to use social media to display good 'grand-childrening' (Barnwell et al., 2021, p. 15) in accordance with common expectations [START_REF] Gorea | Becoming your "authentic" self: How social media influences youth's visual transitions[END_REF]. Indeed, during the pandemic positive feelings towards grandparents (i.e. affectual solidarity) increased when college-aged grandchildren had more frequent contact with their grandparents (i.e.

associational solidarity), even when an increase in contact was partly motivated by the perceived responsibility of grandchildren to do so (McDarby et al., 2021). Grandchildren displayed their grandparents both as 'badass' (for instance when displaying them as funloving dancers (i.e. associational solidarity)) and in need of care, in line with findings from a study on Instagram [START_REF] Barnwell | Captured and captioned: Representing family life on Instagram[END_REF]. Grandparents, on the other hand, seemed less sensitive to feelings of increased responsibility towards their grandchildren during the pandemic. Moreover, grandparents might use TikTok as a platform to be the voice of their generation [START_REF] Brewer | Tell it like it really is": A case of online content creation and sharing among older adult bloggers[END_REF] about their role as grandparents during the pandemic, as a counter-discourse of ageist reports. Grandparents and grandchildren, thus, connect a different set of personal experiences to a more generalized understanding of family relationships [START_REF] Finch | Displaying families[END_REF] which leads to contrasting displays of grandparent-grandchild relationships on TikTok during the COVID-19 pandemic.

Limitations and Future Directions

The current study is the first one to qualitatively study grandparent-grandchild intergenerational solidarity on TikTok. Our results expand previous work on young TikTok users across generations and highlight both differences and commonalities between grandparents and grandchildren on TikTok. Comparative studies not only combat ageist stereotypes regarding digital practices of older people, but they also provide novel insights on young people beyond peer-to-peer interactions. Similar efforts on social media platforms are meaningful to study both in the scope of ISF and family display beyond the COVID-19 pandemic. Notwithstanding our intention to include both grandparents and grandchildren in our study, the display of intergenerational solidarity between grandparents and grandchildren on TikTok is dominated by the grand-children. Even though they are likely to post idealized content [START_REF] Barnwell | Captured and captioned: Representing family life on Instagram[END_REF] our results emphasize experiences and intentions of the grandchildren. Also, the current analysis is restrained by the language barrier, which may restrict the cultural diversity of the videos included in the data corpus.

Each video is a brief overview of what TikTok users want to show to their community.

It does not allow us to understand the context of the video, the intentions of the video creators or the shared meaning-making processes between the family members who interact with each other on TikTok. Further studies could use interviews to focus on grandparents and grandchildren, and inquire into motivations, doubts and experiences related to how they display family relationships on TikTok. Of special interest are studies that focus both on those who make and post videos, and those who are featured in videos to understand both intentions and effects. Additional analysis of the user comments on TikTok would foster possibilities to understand the impact of the representation of grandparents and grandchildren (or older people and youth in general) on this platform.

Conclusion

This paper offers an account of the display of intergenerational solidarity between grandparents and grandchildren on TikTok during the COVID-19 pandemic. The results suggest that grandparents and grandchildren value qualities of physical touch and family celebrations (i.e. affectual solidarity), and big life events (i.e. normative solidarity) in their relationship, as they put these on display to other users on TikTok. We also found that grandparents and grandchildren participate in platform-initiated activities, like dance challenges. Although grandparents and grandchildren refrain from consensual solidarity on TikTok, the results in other ISF categories provide clues to differences in the public display of their respective roles. Grandchildren were found to emphasize their role in protecting their grandparents when remaining outdoors (i.e. associational solidarity), taking extra sanitary measures (i.e. normative solidarity), or preparing meals and care packages for their grandparents (i.e. functional solidarity). Such differences emphasize the different experiences and display needs of grandparents and grand-children during the COVID-19 pandemic on TikTok.

Figure 1 .

 1 Figure 1. Flow diagram of the selection process.

  the different ways in which grandparents and grandchildren displayed intergenerational solidarity on TikTok during the COVID-19 pandemic. The data corpus yielded most entries for affectual solidarity, illuminating how grandparents and grandchildren express their affection for each other. Associational solidarity was the second most prominent dimension, suggesting that grandparents and grandchildren found several ways to share common activities during the pandemic. The third largest category was functional solidarity, with videos showing how grandparents and grandchildren helped each other in a difficult time. The following sections present an overview of all six dimensions of the Intergenerational Solidarity Framework (ISF).

Figure 2 .

 2 Figure 2. Grandparents and grandchildren hug, while grandchildren are dressed in full body suits and wearing face masks.

Figure 3 .

 3 Figure 3. Grandfather and granddaughter hug through a plastic wall.

Figure 4 .

 4 Figure 4. Grandparents and grandchildren stand in separate rows to perform the Laxed Siren Beat dance challenge which only include arm movements so grandparents can sit.

  14 videos, grandparents (n = 1) and grandchildren (n = 13) did chores for each other. Chores included doing groceries (mainly grandchildren shopping for grandparents), grandchildren painting a shed in their grandparents' garden (n = 2) or gardening, a grandfather installing a bookshelf, a granddaughter cleaning up or a grandchild shaving a goat. Another type of instrumental support was technical support (n = 4) provided by grandchildren to their grandparents. These videos included checking the Wi-Fi connection, dropping off a tablet streaming the Sunday mass live, or helping out grandparents during video calls (n = 2). Other videos displayed emotional support (n = 4) to cheer up grandparents or grandchildren (e.g., grandparents cheering a baby who started crawling, grandchildren helping their grandparents to engage in an activity in order to prevent boredom such as setting up a Netflix or TikTok account or lending a pet for a sleepover). The last example of instrumental support is displayed through one video of a grandson offering physical support to his grandparents over distance by showing them different physical exercises during a video call from his bedroom.

  measures used in these videos were physical distance (n = 78), wearing a mask (n = 35), interacting through a window (n = 27), wearing gloves (n = 7), and wearing full-body suits (n = 6). Interestingly, on videos shared by grandchildren, 46 were not eligible to report the use of barrier measures but on the other 141 videos, 65.96% showed the use of barrier measures in the interaction with grandparents and 34.04% did not show any kind of barrier measures. On the opposite, on the 70 videos shared by grandparents, if 19 were not eligible to report the use of barrier measures, on the other 51 videos, 84.31% did not show any use of protection during interactions with grandchildren and only 15.68% showed the use of barrier measures (wearing a mask = 4; physical distance = 3; interacting through a window = 2).

Table 2 .

 2 Coding scheme following the ISF.

	Affectual	Associational	Consensual	Functional	Normative	Structural
	Affectionate	TikTok	Politics	Instrumental	First grandchild	Health
	gestures	Challenge	Sexuality	support	Pregnancy	Travelling
	Celebration	Keeping		Food from GP	announcement	Household
	and gift	entertained		to GC	Favorite
	Fun and humor	Door visit		Food from GC	grandchild
	Art and cards	Garden visit		to GP	Protecting GP
	Grief	Beauty salon		Non-essential
	Memories	Phone and video		items
	Missing each	calls
	other	Baking together
		Accidental
		meeting


Nicolas Puchot

Marie-Thérèse Lecoq

Romuald Carinci

Jean Bernard Duchemin

Mathilde Gendrin

Catherine Bourgouin
email: catherinebourgouincabourg@pasteur.fr

Marie-The ´rèse Lecoq

Joao Pinto

Thomas Walker

Establishment of a colony of Anopheles darlingi from French Guiana for vector competence studies on malaria transmission

Keywords: Anopheles darlingi, colony, rearing, Plasmodium, susceptibility, French Guiana

ou non, émanant des établissements d'enseignement et de recherche français ou étrangers, des laboratoires publics ou privés.

Introduction

The establishment of mosquito colonies has been instrumental for a better understanding of many of their biological features including development, insecticide resistance, and pathogen transmission. It is also important to test novel insecticides and novel technologies for population suppression or replacement such as the sterile insect technique, the RIDL approach (Release of Insects carrying Dominant Lethals), the use of symbiont bacteria such as Asaia, or the production of transgenic mosquitoes.

Among mosquito species known as vectors of human pathogens, Anopheles species involved in malaria parasite transmission are the more numerous and diverse worldwide compared to mosquito species involved in arbovirus transmission [START_REF] Sinka | A global map of dominant malaria vectors[END_REF]. In addition, Anopheles species have specific reproductive characteristics, notably their eurygamous behavior, i.e., their propensity to mate in swarms and the female refractoriness to multiple mating. Those characteristics have been challenging for the successful establishment of colonies of many Anopheles species. To overcome these limitations, one strategy is the forced mating technique, which relies on the manual adjustment of male and female copulatory structures [START_REF] Baker | Induced copulation in Anopheles mosquitoes[END_REF][START_REF] Ow Yang | Maintenance of a laboratory colony of Anopheles maclalatus (Theobald) by artificial mating[END_REF][START_REF] Baker | Mating problems as related to the establishment and maintenance of laboratory colonies of mosquitos[END_REF]. Although tricky and highly demanding in human resources, this technique was successful in maintaining some species as laboratory colonies for many years. The methodology has notably been used to colonize and maintain Anopheles dirus and is still used to create and maintain novel Anopheles colonies [START_REF] Amir | Colonization of Anopheles cracens: a malaria vector of emerging importance[END_REF]. Interestingly, some species such as Anopheles albitarsis were reported to change to stenogamous free-mating behavior after several generations of colony maintenance by forced copulation [START_REF] Horosko | Establishment of a free-mating colony of Anopheles albitarsis from Brazil[END_REF].

Anopheles darlingi [also called Nyssorhynchus darlingi [START_REF] Marinotti | What is in a name? Anopheles darlingi versus Nyssorhynchus darlingi[END_REF]] is a major vector of both Plasmodium falciparum and Plasmodium vivax in South and Central America including French Guiana [START_REF] Conn | Chapter 5 -Ecology of Anopheles darlingi, the primary malaria vector in the americas and current nongenetic methods of vector control[END_REF][START_REF] Sinka | The dominant Anopheles vectors of human malaria in the americas: occurrence data, distribution maps and bionomic precis[END_REF]. As most anophelines, An. darlingi has a eurygamous behavior that precludes its readiness to mate in captivity. In contrast to many Anopheles species belonging to the Cellia and Anopheles subgenera, it is not clear whether Anopheles belonging to the Nyssorhynchus subgenus exhibit a swarming behavior for mating [START_REF] Yuval | Mating systems of blood-feeding flies[END_REF]. Indeed, Lounibos et al. reported that young F1 virgin An. darlingi females would mate within 2 to 36 h after being released in a village where no mosquito swarm was evidenced [START_REF] Lounibos | Prompt mating of released Anopheles darlingi in western Amazonian Brazil[END_REF]. In 61cm-high cages, Villarreal-Treviño et al. observed pseudo-swarms of An. darlingi occurring 5-40 cm above the cage floor [START_REF] Villarreal-Trevino | Establishment of a free-mating, longstanding and highly productive laboratory colony of Anopheles darlingi from the Peruvian amazon[END_REF]. This may be due to space constraints yet corroborates field observations that males fly at low altitude. As An. darlingi is a major malaria vector in South America and particularly in the Amazon basin, several laboratories tried to colonize this mosquito species for vector competence studies and additionally for human Plasmodium sporozoite production for vaccine development studies. The oldest published records of the establishment of a free-mating An. darlingi colony go back to 1947 with An. darlingi from the Co-operative Republic of Guyana (at that time British Guiana), Colombia, and Brazil [START_REF] Giglioli | Laboratory colony of Anopheles darlingi[END_REF][START_REF] Bates | The laboratory colonization of Anopheles darlingi[END_REF][START_REF] Freire | Rearing and some data on the biology of A. darlingi[END_REF]. Since then, two reports of Brazilian (Araraquara, Sao Paulo State) An. darlingi colonies have been published in 1970 and 1988 [START_REF] Correâ | Informe sobre uma colonia de anopheles darlingi[END_REF][START_REF] Buralli | Maintenance of Anopheles darlingi root, 1926 colony, in the laboratory[END_REF] and followed almost 40 years later with the description of three novel free-mating An. darlingi colonies, two from Peru and one from the Rondonia State (Amazon basin-Brazil) [START_REF] Villarreal-Trevino | Establishment of a free-mating, longstanding and highly productive laboratory colony of Anopheles darlingi from the Peruvian amazon[END_REF][START_REF] Moreno | Infection of laboratory-colonized Anopheles darlingi mosquitoes by Plasmodium vivax[END_REF][START_REF] Araujo | Brazil's first free-mating laboratory colony of Nyssorhynchus darlingi[END_REF].

Here, we describe the establishment of a free-mating colony of An. darlingi from a different South American country, French Guiana. By mixing our expertise in Anopheles mosquitoes with published results and improvements along the most recent established Anopheles pseudopunctipennis and An. darlingi free-mating colonies [START_REF] Villarreal-Trevino | Establishment of a free-mating, longstanding and highly productive laboratory colony of Anopheles darlingi from the Peruvian amazon[END_REF][START_REF] Moreno | Infection of laboratory-colonized Anopheles darlingi mosquitoes by Plasmodium vivax[END_REF][START_REF] Lardeux | Laboratory colonization of Anopheles pseudopunctipennis (Diptera: Culicidae) without forced mating[END_REF][START_REF] Villarreal | Colonization of Anopheles pseudopunctipennis from Mexico[END_REF], this novel An. darlingi colony was produced in less than 8 months and has currently reached its 21st generation.

Materials and methods

Producing the F1 generation from wild Anopheles darlingi Wild An. darlingi were captured in Val'ranch (Tonate Macouria village; 05.01616°N 052.52845W), a location where this species is abundant [START_REF] Vezenegho | Anopheles darlingi (Diptera: Culicidae) dynamics in relation to meteorological data in a cattle farm located in the coastal region of French Guiana: Advantage of mosquito magnet trap[END_REF], using two Mosquito Magnet ® MM300 Executive traps arranged 50 m apart. The capture was performed over a single night with two catching sessions from 6 p.m. to 8 p.m., and from 8 p.m. to 7 a.m. in September 2020. Female mosquitoes were transferred from the traps into 30-cm 3 plastic cages (Bugdorm ® ) covered with damp towels to maintain high humidity. They were transported to the laboratory (Institut Pasteur de la Guyane, Cayenne, French Guiana) in a foam box to buffer ambient temperature. There, they were fed on anesthetized mice. On the fourth day after feeding, one wing was removed from gravid females to induce oviposition, and the females were transferred to individual Eppendorf tubes containing wet filter paper as an additional forced-egg laying method [START_REF] Nepomichene | Efficient method for establishing F1 progeny from wild populations of Anopheles mosquitoes[END_REF]. Once the eggs were laid, the females were removed before air freight transfer of tubes with egg clutches to Institut Pasteur (Paris, France). Initial An. darlingi identification relies on adult morphology characteristics and is further confirmed by COI DNA barcoding on adults from the 21st generation, using the CO1 universal primers designed for invertebrates by Folmer et al. [START_REF] Folmer | DNA Primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates[END_REF] (Supplementary Data).

Insectary parameters

Following the publications on An. pseudopunctipennis [START_REF] Lardeux | Laboratory colonization of Anopheles pseudopunctipennis (Diptera: Culicidae) without forced mating[END_REF] and An. darlingi [START_REF] Villarreal-Trevino | Establishment of a free-mating, longstanding and highly productive laboratory colony of Anopheles darlingi from the Peruvian amazon[END_REF][START_REF] Moreno | Infection of laboratory-colonized Anopheles darlingi mosquitoes by Plasmodium vivax[END_REF], the light periodicity was set up at 12:12 with dawn and dusk spanning each 1 h. To stimulate copulation, blue light flashes (LED stroboscopic light) were provided for 45 min just after full darkness at a rate of 140 flashes per minute, each flash lasting ∼0.2 s. The temperature in French Guiana cycles daily between 22 and 32°C when An. darlingi can be found (Supplementary Figure 1, https://donneespubliques. meteofrance.fr/?fond=produit&id_produit=90&id_rubrique= 32-; access date 23 August 2019). Considering these natural temperature conditions and expecting an adapted behavior keeping mosquitoes away from high temperatures in the field, the insectary temperature was set at 26°C during the daytime and down to 23°C during nighttime. A constant relative humidity was kept at approximately 75%, which is close to the average humidity observed in the field during An. darlingi peaks (81%, Supplementary Figure 1). A summary of those experimental conditions is presented in Figure 1.

Mosquito rearing

Larvae were reared in white square plastic pans filled to 2 cm height with demineralized water supplemented with a mixture of salts (200× solution: NaCl 0.2%; CaSO 4-2H 2 O 0.3%; NaHCO 3 1%) and covered with a mosquito net. L1 to L2 larvae were fed with Tetra ® TetraMin Baby and in later stages with cat food. When the first pupae appeared, a small container filled with a sterile 10% sucrose solution in mineral water (Volvic ® ) and holding a dental cotton roll (ROEKO Parotisroll Coltene-10 cm height) was placed in the center of the rearing tray to feed emerging adults. Adults were collected with an in-house made electric aspirator and transferred to mating cages (47.5 × 47.5 × 47.5 cm; BugDorm-4F4545-https://shop.bugdorm.com). Because of the size of the cage, two sucrose solution containers were placed at different heights in the cage. They were replaced every other day.

At each generation, mosquitoes were given 5 to 7 days for free mating (with light-flash induction, as described above) before being fed on anesthetized animals, either rabbit or mice (see details in Results).

Females were allowed to lay eggs in a black plastic tray filled to 1.5 cm height with a 2:1 mix of demineralized water and filtersterilized water (22-µm filters) from the larval containers as a possible attractive cue to stimulate females to deposit their eggs. Eggs were collected by filtration through an in-house made strainer equipped with Nitex (125-µm fine mesh-Dutscher #987452) and then transferred for hatching in a rearing pan.

Experimental infections with Plasmodium berghei and Plasmodium falciparum

Five-day-old An. darlingi females were infected with P. berghei (GFP ANKA@HSP70 clone) or P. falciparum NF54, concurrently with same-age An. stephensi females (Sda500 strain). The mosquito infections were performed by the local Anopheles infection platform (CEPIA, Institut Pasteur). Before being exposed to an infected blood meal, females were starved from sugar overnight with access to water only in the form of a damp cotton ball, which was removed 4 h before blood feeding. Mosquito infection with P. berghei was performed by placing two anesthetized infected mice on top of the cage for 15 min. Only fully fed females were maintained for 10 days at 21°C for oocyst development. Mosquito infection with P. falciparum was performed by standard membrane feeding assay (SMFA) using glass feeders containing in vitro produced gametocytes as described previously [START_REF] Mitri | Density-dependent impact of the human malaria parasite Plasmodium falciparum gametocyte sex ratio on mosquito infection rates[END_REF]. Fully fed females were transferred to small cages and maintained for 7 days (oocyst detection) at 26°C.

Oocysts were detected on dissected midguts stained with 0.1% mercuro-bromo fluorescein (Fluka) in 1× phosphate buffered saline.

Animals-Ethics statement

This study was conducted in strict accordance with the recommendations from the Guide for the Care and Use of Laboratory Animals of the European Union (European Directive 2010/63/UE) and the French Government. The

Results

Establishing the first six generations of free-mating An. darlingi

Among 120 caught wild An. darlingi females, 54 took a blood meal on mice, and 34 produced eggs. From these 34 wild An. darlingi females, we obtained ~2,000 F1 eggs that led to 705 adults with a 1:1 sex ratio. Seven days after the first F1 adults emerged, mosquitoes were fed on anesthetized animals for nine consecutive days followed by four additional feedings spaced roughly every 3 days (Figure 2). To avoid mosquitoes drowning in the water of the egg-laying container, this was provided to females after the fifth blood meal only. The container was inspected every day for egg laying and the first eggs were laid after the sixth blood meal. Only 20 eggs were collected on that day. Although eggs were collected almost every day since the sixth blood meal, the size of each egg clutch rarely exceeded 50 eggs. Nevertheless, combining the frequency of blood feedings and egg collection over nearly 3 weeks allowed us to collect ~1,050 F2 eggs that produced 505 adults. The same procedure and blood feeding scheme (Figure 2) allowed us to reach an F6 population of nearly 3,300 adults without any trouble (Figure 3).

Bionomic features Egg production and development

The wild-caught females produced a mean of 59 eggs per blood-fed female (Table 1). This number dropped between an estimated 3 to 13 eggs per female during the establishment of the colony despite providing over nine blood meals to the females (Table 1 and Supplementary Data S1). This low number likely represents the low number of fertilized females and not the intrinsic fecundity of fertilized females. This could be confirmed by additional investigation as spermatheca dissection and single female egg collection.

Eggs laid on the same day after a blood meal hatched over 2 to 7 days leading to larval stages overlapping from L1 to L3. The hatching rate was relatively stable and rarely exceeded 50% (Table 1 and Figure 4), suggesting that unfertilized blood-fed females can lay unfertilized eggs or that the number of spermatozoids transferred to the female during free mating is not sufficient to fertilize all the oocytes developing in a single blood-fed female.

Larval and pupal development

The time between egg laying and first adult emergence was between 14 and 16 days under our insectary settings. The mortality rate of aquatic stages of An. darlingi was estimated as the ratio between the total number of adults and the total number of young larvae (L1 to L2) at each generation. As shown Schematic representation of the production of the first six generations of An. darlingi. The duration of egg hatching, larval stage, and adult stage are color coded. The red line is a schematic representation of the blood feeding strategy that was used over the first six generations by alternate feeding on rabbit (red) and mice (brown). Some additional blood feedings on mice were only provided to the first generation (blue). Between F2 and F6, the number of feedings varied based on the availability of the animals and the egg production and are detailed in Figure 3. At each generation, the time required for the females to lay the first egg batch (green) diminished across generations.

in Figure 5, the aquatic stage mortality rate was generally lower than 25%, except in F4 where it reached 42%. The proportion of females remained stable between 48% and 56% from F1 to F4, indicating that the sex ratio was not distorted from the expected 1:1 ratio (see Table 1).

Monitoring the effect of the blue light flashes and shift in night temperature on free mating

To test whether the blue light flashes and the shift in night temperature were mandatory to maintain the colony as free mating after the first five generations, part of the F5 adults, thereafter called F5', were moved to a similar insectary without any light flashes and with constant temperature (26°C) producing the F6', while the remaining F5 adults were maintained with blue light flashes and low night temperature when producing the F6 generation. The F7 and F7' generations were obtained from F6 and F6', respectively. Conversely, the next two generations from the F7' parents were submitted to light flashes and maintained at a constant temperature (26°C).

As presented in Table 2, we did not detect any significant differences in the egg production between F and F' at each generation, whether or not normalizing data to the number of blood feedings each generation received (Supplementary Data S2; Wilcoxon test on eggs/female-p = 1; on egg/female/blood feed-p = 1). This suggests that past the fifth generation, neither light flashes nor a shift in night temperature is required for maintaining this free-mating An. darlingi colony. Some F13 eggs were sent to Institut Pasteur de la Guyane (Cayenne, French Guiana) to rear a duplicate colony. In both laboratories, generation F21 is currently ongoing.

An. darlingi permissiveness to P. berghei and P. falciparum infection

We then tested whether this An. darlingi colony could be a valuable tool for vector competence studies. To this aim, An. darlingi and An. stephensi females were first infected in parallel The calculation of eggs/female for the F6 and F7 generations used a 1:1 sex ratio to derive the number of females from the count of total adult mosquitoes. ND, Not Determined. Yield in eggs and adults from F1 to F6. The graph represents the number of eggs obtained at each generation (blue line with squares and left scale) and the subsequent number of adults (red curve with circles and right scale). The number of blood feedings provided at each generation is indicated below.

with P. berghei. None of the 22 blood-fed An. darlingi females developed any oocyst, whereas 75% of the blood-fed An. stephensi females (n = 16) were infected with a mean oocyst load of 85.5 oocysts per infected female (range 27-100; Figure 6A). This is the first report revealing the possible refractoriness of An. darlingi to a rodent malaria parasite. Next, An. darlingi and An. stephensi females were infected with the same batch of in vitro produced P. falciparum gametocytes. Across three independent infection experiments, An. darlingi was found susceptible to P. falciparum NF54 (Figure 6B). However, both infection prevalence and oocyst load were lower than the ones of An. stephensi.

Discussion

Here, we report on the successful establishment of a freemating An. darlingi colony starting from wild-caught females in French Guiana. An. darlingi has a wide geographic distribution from Central America to many countries in South America including the diverse regions of the Amazon basin where this species was identified as a dominant malaria vector [in [START_REF] Sinka | A global map of dominant malaria vectors[END_REF]]. Based on various genetic analyses An. darlingi is composed of clearly isolated populations or clades [START_REF] Conn | Chapter 5 -Ecology of Anopheles darlingi, the primary malaria vector in the americas and current nongenetic methods of vector control[END_REF][START_REF] Prado | Behavior and abundance of anopheles darlingi in communities living in the Colombian Amazon riverside[END_REF]. The present colony likely represents a novel genetic resource of An. darlingi that will offer additional values for a better characterization of the genetic Larval survival success to adulthood across the first six generations of An. darlingi. The survival success is visualized from the mortality rate of larvae that did not reach adulthood, estimated from F1 to F6. Egg eclosion rate at each generation. The number of L1 larvae hatching from each egg batch was determined by daily counting. L1 to L2 larvae hatching over the weekend were recorded on Mondays.

diversity and vectorial competence of this major South American malaria vector.

The colony was established from the progeny of 34 wildcaught females. This limited number of founders provided a starting pool of roughly 2,000 eggs. This number of founders is quite low compared to the numbers used for the most recently established An. darlingi colonies, which were initiated from 135 up to 878 wild-caught females [START_REF] Villarreal-Trevino | Establishment of a free-mating, longstanding and highly productive laboratory colony of Anopheles darlingi from the Peruvian amazon[END_REF][START_REF] Moreno | Infection of laboratory-colonized Anopheles darlingi mosquitoes by Plasmodium vivax[END_REF][START_REF] Araujo | Brazil's first free-mating laboratory colony of Nyssorhynchus darlingi[END_REF].

To establish this colony, we developed a pragmatic approach by pooling information from our preliminary experiments and from published records on An. darlingi colonization since 1947. These data indicated that female insemination was a key

B A FIGURE 6
Infection of An. darlingi with P. berghei and P. falciparum. (A) Five-day-old An. darlingi females from the F8 generation, and An. stephensi (Sda 500) were starved overnight from sugar and fed on the same mice harboring P. berghei gametocytes. (B) Five-day old An. darlingi females and An. stephensi (Sda 500) were starved overnight from sugar and fed on P. falciparum NF54 gametocytes. Three experimental infections were performed using F9, F10, and F13 An. darlingi females, corresponding to Experiments 1, 2, and 3, respectively. The horizontal bar on the graph represents the median of developing oocysts per mosquito midgut. Detailed data on prevalence and infection intensity are indicated below. Mean oocyst counts and data range derive from infected midguts only. parameter and that forced mating did not allow fertilization, even though this technique was easy to perform with An. darlingi ( [START_REF] Villarreal-Trevino | Establishment of a free-mating, longstanding and highly productive laboratory colony of Anopheles darlingi from the Peruvian amazon[END_REF][START_REF] Moreno | Infection of laboratory-colonized Anopheles darlingi mosquitoes by Plasmodium vivax[END_REF][START_REF] Lardeux | Laboratory colonization of Anopheles pseudopunctipennis (Diptera: Culicidae) without forced mating[END_REF], data not shown). We combined two main strategies. The first one capitalized on recent data obtained with An. pseudopunctipennis [START_REF] Lardeux | Laboratory colonization of Anopheles pseudopunctipennis (Diptera: Culicidae) without forced mating[END_REF][START_REF] Villarreal | Colonization of Anopheles pseudopunctipennis from Mexico[END_REF] and An. darlingi [START_REF] Villarreal-Trevino | Establishment of a free-mating, longstanding and highly productive laboratory colony of Anopheles darlingi from the Peruvian amazon[END_REF] that suggest that mosquito copulation can be induced by providing light flashes just after dusk and decreasing night temperature, as happens in wild mosquito populations after dusk. The second strategy was to provide repeated blood meals to female mosquitoes up to 14 per generation over the first six generations. Providing light flashes after dusk as a copulation inducer was used for the recently established An. darlingi colonies from Peru and Brazil [START_REF] Villarreal-Trevino | Establishment of a free-mating, longstanding and highly productive laboratory colony of Anopheles darlingi from the Peruvian amazon[END_REF][START_REF] Moreno | Infection of laboratory-colonized Anopheles darlingi mosquitoes by Plasmodium vivax[END_REF][START_REF] Araujo | Brazil's first free-mating laboratory colony of Nyssorhynchus darlingi[END_REF] while decreased night temperature was mentioned in only two reports [START_REF] Villarreal-Trevino | Establishment of a free-mating, longstanding and highly productive laboratory colony of Anopheles darlingi from the Peruvian amazon[END_REF][START_REF] Araujo | Brazil's first free-mating laboratory colony of Nyssorhynchus darlingi[END_REF].

Our data reported in Table 2 show that neither light flashes nor night temperature shift is important past the sixth generation. While we did not assess their role in the first six generations, we observed sexual activity as soon as light flashes started for the F1. In 1947 Bates observed "sexually excited males and mating" after exposure to lights of different colors [START_REF] Bates | The laboratory colonization of Anopheles darlingi[END_REF]. He reported that mating occurred in the presence of a dim blue light when a limited number of mosquitoes (328 adults) were kept in a small cage, while mosquitoes did not mate without artificial light. In our insectary conditions, even if copulation was induced after dusk using light flashes, copulation was also observed in the daytime under dim light past the 10th generation. Similarly, Villarreal-Treviño et al. reported a very strong impact of the light-temperature combination to favor insemination at the 2nd generation while light stimulation was not required anymore after the 10th generation [START_REF] Villarreal-Trevino | Establishment of a free-mating, longstanding and highly productive laboratory colony of Anopheles darlingi from the Peruvian amazon[END_REF]. Because of limited space, we chose middle-size adult cages (47.5 × 47.5 × 47.5 cm) despite data suggesting that larger cages (60 × 60 × 60 cm) or even bigger (1.2 m 2 × 3 m high) do better [START_REF] Villarreal-Trevino | Establishment of a free-mating, longstanding and highly productive laboratory colony of Anopheles darlingi from the Peruvian amazon[END_REF][START_REF] Giglioli | Laboratory colony of Anopheles darlingi[END_REF][START_REF] Bates | The laboratory colonization of Anopheles darlingi[END_REF][START_REF] Freire | Rearing and some data on the biology of A. darlingi[END_REF]. As recommended for Anopheles funestus, another difficult mosquito species to rear, and tested for An. darlingi [START_REF] Villarreal-Trevino | Establishment of a free-mating, longstanding and highly productive laboratory colony of Anopheles darlingi from the Peruvian amazon[END_REF], we used dark containers as egg-laying devices.

Following published data, mosquitoes were given their first blood meal on day 6 or day 7 post-emergence to ensure that the copulation stimuli were efficient enough to produce a reasonable number of fertilized females at each generation. Mosquitoes were then fed almost every day on anesthetized rabbits or mice. Feeding mosquitoes on more than one type of animal blood was reported to favor female fecundity and we previously used this strategy to establish an An. arabiensis colony from Madagascar (Puchot & Bourgouin, unpublished).

As presented in Tables 1 and2 and Supplementary Figures S2 andS3, female fecundity (eggs/female) seems rather low at each generation. These numbers are in the same range as the ones obtained by Villarreal-Trevino et al. [START_REF] Villarreal-Trevino | Establishment of a free-mating, longstanding and highly productive laboratory colony of Anopheles darlingi from the Peruvian amazon[END_REF], using the same calculation method taking into account the entire female population rather than individual counts per fertilized female [START_REF] Moreno | Infection of laboratory-colonized Anopheles darlingi mosquitoes by Plasmodium vivax[END_REF]. It is noticeable that the egg-to-female ratio increased in the eighth generation while decreasing the number of blood meals provided. The number of blood meals has been reduced to six or seven from the 9th generation, and to three from the 13th generation. Meanwhile, we stopped counting the eggs as we had enough to proceed to the next generation.

Regarding the development time from egg to adult, we obtained very similar results to published data in the range of 14 to 16 days [START_REF] Villarreal-Trevino | Establishment of a free-mating, longstanding and highly productive laboratory colony of Anopheles darlingi from the Peruvian amazon[END_REF][START_REF] Giglioli | Laboratory colony of Anopheles darlingi[END_REF]. Noticeable was the asynchronous larval development from the same egg batch. Whether this was due to genetically different embryonic or larval development time or to the quantity and the type of the food provided to the larvae will need further investigation.

Importantly, we demonstrated that our free-mating An. darlingi colony is susceptible to in vitro produced gametocytes from P. falciparum NF54 originating from Africa. By comparison with concomitant An. stephensi control infections, the prevalence of infection was, however, slightly lower than the one obtained with F1 An. darlingi from Belize infected with the same P. falciparum strain [START_REF] Grieco | Comparative susceptibility of three species of Anopheles from Belize, central America, to Plasmodium falciparum (NF-54)[END_REF]. This difference may reflect an overall different genetic background between Belize and French Guiana An. darlingi populations or the selection of genotypes along the establishment of the colony from a limited number of founders. Interestingly, among the other species of the Nyssorhynchus subgenus, Anopheles albimanus is moderately refractory to P. falciparum NF54, whereas it is more permissive to P. falciparum 7G8, originating from Brazil [START_REF] Molina-Cruz | Plasmodium evasion of mosquito immunity and global malaria transmission: The lock-and-key theory[END_REF]. By contrast Anopheles aquasalis is refractory to both P. falciparum strains unless its immune system was affected by silencing the mosquito complement factor LRIM1 (Leucin-Rich-repeat-containing IMmune gene 1) or antibiotic treatment to remove their microbiota [START_REF] Orfano | Plasmodium yoelii nigeriensis (N67) is a robust animal model to study malaria transmission by south American anopheline mosquitoes[END_REF]. When challenged with P. berghei, our colony showed full refractoriness, as observed for An. aquasalis [START_REF] Orfano | Plasmodium yoelii nigeriensis (N67) is a robust animal model to study malaria transmission by south American anopheline mosquitoes[END_REF], suggesting that this parasite species is not a suitable model to study An. darlingi-Plasmodium interactions. Further investigation would be necessary to determine whether P. berghei is developing more slowly in An. darlingi, as previously shown in An. albimanus [START_REF] Frischknecht | Using green fluorescent malaria parasites to screen for permissive vector mosquitoes[END_REF], and whether Plasmodium yoelii is a suitable rodent malaria model in An. darlingi, as demonstrated in An. albimanus [START_REF] Orfano | Plasmodium yoelii nigeriensis (N67) is a robust animal model to study malaria transmission by south American anopheline mosquitoes[END_REF].

In conclusion, this novel An. darlingi colony established from French Guiana mosquitoes offers an invaluable tool for a better understanding of An. darlingi genetic diversity, physiology, and vector competence towards South America malaria parasites including P. falciparum and P. vivax.

Data availability statement

The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.

FIGURE 1

 1 FIGURE 1 Insectary parameters. The graph represents temperature (green), relative humidity (blue), and light intensity (red) recorded via a Hobo 1019 recorder (MX1104). Temperature and humidity parameters are provided by a dedicated automated system. Light (LED) is controlled by an electronic device. Blue light flashes are provided by a LED stroboscopic light, just after dusk (red asterisk).

FIGURE 2

 2 FIGURE 2

FIGURE 3

 3 FIGURE 3

FIGURE 5

 5 FIGURE 5

FIGURE 4

 4 FIGURE 4

TABLE 1

 1 This table summarizes the raw data obtained across 7 free mating generations of An. darlingi, starting with 34 wild caught gravid females producing 2000 F1 eggs that led to the production of 705 F1 adults.

	Parental	Number	Number	Numbers	Numbers	Number of	egg eclo-	Larvae to adults	Eggs/	Eggs/female/
	Generation	of eggs	of larvae	of females	of adults	blood meals	sion rate	mortality (%)	female	blood meal
							(%)
	F0			34	34	1			58.8	58.8
	F1	2000	800	344	705	14	40	11.88	3.0	0.2
	F2	1045	530	247	505	11	51	4.72	11.0	1.0
	F3	2705	1425	590	1099	9	53	22.88	5.6	0.6
	F4	3300	1780	578	1040	10	54	41.57	11.7	1.2
	F5	6750	3029	ND	2605	9	45	14.00	5.4	0.6
	F6	7390	3862	ND	3351	7	52	13.23	5.9	0.8
	F7	9650	ND	ND	4097	7	ND	ND	8.7	1.2

TABLE 2

 2 Effect of the blue light flashes and shift in night temperature on free mating of An. darlingi.At generations 5-8, the number of eggs/female is not affected by light flashes and temperature shifts (Wilcoxon test F vs F'-p=1).To keep similar numbers of adults between the F and F' generations we mixed F and F' parents in three instances (F6', F7' and F8). ND, Not Determined.

	Parental	Generation	Number of	Number of	Number	Number of	Eggs/	Eggs/female/	Insectary
	generation		adult parents	blood feedings	of eggs	adults	female	blood meal	conditions
	F5 parents	F5	2006	9	5400	2806	5.4	0.6	flash light and
									23°C night
		F5'	600	8	1990	545	6.6	0.8	no flash light, 26°C
									constant
	F6 Parents	F6	2000	7	5900	2669	5.9	0.8	flash light and 23°C
									night
	545 F6' + 806	F6'	1351	6	3750	1428	5.6	0.9	no flash light, 26°C
	F6								constant
	F7 Parents	F7	2019	7	8750	1760	8.7	1.2	flash light and 23°C
									night
	1428 F7' + 500	F7'	2078	7	7255	2073	7.5	1.1	flash light, 26°C
	F7								constant
	1760 F8 + 142	F8	1902	6	> 10,000	ND	10.5	1.8	flash light and 23°C
	F8'								night
	F8' Parents	F8'	1932	6	> 10,000	ND	10.4	1.7	flash light, 26°C
									constant

Frontiers in Tropical Diseases frontiersin.org

Puchot et al. 10.3389/fitd.2022.949300 Frontiers in Tropical Diseases frontiersin.org

Acknowledgments

We thank members of the CEPIA platform for infecting our mosquitoes (An. darlingi and An. stephensi) with P. berghei and P. falciparum. Thanks to current and previous members of the Vectopole, especially Pascal Gaborit for technical help and to Anavaj Sakuntabhai for his constant support.

Funding

This work was supported by funds from the LabEx IBEID (The Laboratory of Excellence (LabEx) Integrative Biology of Emerging Infectious Diseases-grant no. ANR-10-LABX-62-IBEID) to CB and MG, FEDER-Controle to JD and RC, ANR JCJC MosMi to MG (grant no. ANR-18-CE15-0007), and a Calmette-Yersin Fellowship from Pasteur Network to RC. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Author contributions

Conceived the project and funding: CB, JD, and MG. Designed the experiments: NP and CB. Performed the experiments: NP, M-TL, and RC. Analyzed the data: NP and CB. Drafted the paper: CB and NP. Reviewed and edited the manuscript: NP, JD, MG, and CB. All authors contributed to the article and approved the submitted version.

Conflict of interest

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

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Supplementary material

The Supplementary Material for this article can be found online at: https://www.frontiersin.org/articles/10.3389/ fitd.2022.949300/full#supplementary-material