DOI: 10.5061/dryad.bg79cnpfm
title: Influence of endosymbionts on the reproductive fitness of the tick Ornithodoros moubata tested through elimination of the microbiota using antibiotic treatments

description: Over the past ten years, many studies demonstrated the crucial role of the
 tick microbiome in tick biology. The soft tick Ornithodoros moubata is a
 hematophagous ectoparasite of Suidae particularly known to transmit the
 African swine fever virus. Its bacterial microbiota is characterized by a
 high prevalence of Francisella-like and Rickettsia endosymbionts. The
 present study aims to better understand the potential influence of the
 microbiota on the reproductive fitness of O. moubata. A total of 132 adult
 female ticks were treated using gentamycin or rifampicin added to the
 blood meal. Half of the ticks also received a supplementation with B
 vitamins to address the nutritional role of endosymbionts. Over two
 periods of 50 days, several traits related to reproductive fitness were
 monitored to investigate the importance of Francisella and
 Rickettsia for those traits. It appeared that most of the considered
 reproductive parameters were not affected. However, antibiotic treatments
 induced an increase in the tick survival indicating a potential fitness
 cost of harboring endosymbionts for ticks during the reproduction period.
 Similarly, 366 first-stage nymphs of Ornithodoros moubata were exposed to
 the same treatments for molecular quantification of both endosymbionts.
 Results from qPCR suggested that treatments produced a bacteriostatic
 effect on endosymbionts without fully eliminating Francisella or
 Rickettsia.
description: This dataset was collected over
 two different experiments (see manuscript
 https://doi.org/10.1101/2023.05.09.539061 for more details). 
 The subfile
 "ReproductionTraits" contains the data issued from monitoring of
 life history traits in arthropods treated or not with antibiotics and/or B
 vitamins : Antibiotics
 and B vitamins were added in a blood meal of adult female ticks
 <em>Ornithodoros moubata</em>. Six different treatment
 conditions were tested: No antibiotic with or without B vitamins;
 Gentamycin with or without B vitamins; Rifampicin with or without B
 vitamins. Concentrations were: gentamycin 10
 µg.mL<sup>-1</sup>, rifampicin 10
 µg.mL<sup>-1</sup>, and B vitamins: a mix of biotin 1
 µg.mL<sup>-1</sup>, folic acid 30
 µg.mL<sup>-1</sup>, riboflavin 20
 µg.mL<sup>-1</sup>. Antibiotics and B vitamins were replaced
 by the same volumes of Milli-Q water for negative controls (No antibiotics
 / No vitamins). After
 blood feeding with treatments and being mated with males, the females were
 monitored during two periods of 50 days. We registered survival and traits
 associated with reproduction: laying success, laying date, number of eggs
 laid (standardized with the female tick weight as big ticks tend to lay
 more eggs), egg mass, date of hatching, hatching rate, date of molting
 into first nymphal stage, and molting rate. The data obtained are made available
 here as well as the R script used for statistical analyses. 
   The subfile "qPCR" contains
 the data issued from qPCR performed on arthropods treated or not with
 antibiotics and/or B vitamins : First nymph ticks <em>Ornithodoros
 moubata</em> received a bloodmeal treated with antibiotics and/or B
 vitamins with the same protocol as presented above. Nymphs were then
 sampled at day 0 (before blood meal), day 1, day 3, day 5, day 10, day 15
 and day 30 after blood meal and crushed for DNA extraction. Quantitative
 PCRs were then performed on the extracted DNA to amplify target genes:
 <em>RpoB</em> gene marker of the presence of
 <em>Francisella</em>; <em>GltA</em> gene marker of
 the presence of <em>Rickettsia</em>;
 <em>Actin</em> gene of <em>Ornithodoros
 moubata</em> to assess the success of the extraction and qPCR, and
 to standardize for the tick size. Samples with a qPCR cycle threshold (Ct)
 beyond 38 were considered as negative. For positive samples, Cts were
 converted into concentrations from the standard curves. Ratio of the
 concentration of the gene of interest (GltA or RpoB) divided by the
 concentration of the actin gene was calculated to standardize for the tick
 size. The data obtained
 and conversion performed are made available here as well as the R script
 used for statistical analyses.
description: Data files are in .csv format with
 semicolon as separators, they can be visualized using any software capable
 of handling .csv files (Excel, LibreOffice Calc, TextPad etc…).
 R Scripts require R version 4.2.3
 (2023-03-15) to be launched and can be used easily with RStudio.
description: # Influence of endosymbionts on the reproductive fitness of the tick
 Ornithodoros moubata tested through elimination of the microbiota using
 antibiotic treatments: *** This README document presents the two datasets
 made available here: "qPCR" and "ReproductionTraits"
 ## qPCR - summary The 7 datasheets made available here result from two
 campaigns of data collection and have been adjusted to be used into the R
 script script\_analysis\_of\_qPCR\_results.R available in the same
 archive. This script can be launched using R version 4.2.3 and R studio
 and by installing the packages associated with the library used in the
 script (see ### packages and library ###). All datasheets can be extracted
 in the same folder from the archive qPCR.zip and used directly in the
 script after adding the path of the folder in the script. Data are issued
 from qPCR performed on soft ticks Ornithodoros moubata (first nymphs or
 adults) treated or not with antibiotics and/or B vitamins to eliminate
 their microbiota. Quantitative PCRs were performed to amplify target
 genes: RpoB gene marker of the presence of Francisella; GltA gene marker
 of the presence of Rickettsia; Actin gene of Ornithodoros moubata to
 assess the success of the extraction and qPCR, and to standardize for the
 tick size. ## qPCR - Description of the Data and file structure Data were
 collected over two series of qPCR performed by two different experimenters
 due to material constraints. Indeed, shortage of laboratory supply during
 COVID19 pandemics led to first analyze a small part of the samples (first
 experimenter): Nymphs from day0, day1, day3 and day10 from the treatment
 groups (control, gentamycin, and rifampicin) which were not treated with
 vitamins. All other nymph groups and adults were analyzed later by a
 second experimenter. For qPCR cycle threshold, a threshold &gt;38 was
 considered negative. If the threshold was negative for actin, the qPCR
 result for the sample was considered a false negative and was launched a
 second time. The file names indicate which individual are concerned
 (nymphs or adults) and which genes are studied. Here is the detail of how
 the information were collected/registered/transformed for each variable:
 groupe\_antibiotique: Antibiotic treatment group of the tick (Rifampicin
 R, Gentamycin G or Control T). groupe\_vitamine: Vitamin supplementation
 group of the tick (Vitamin Vit or NoVitamin NoVit). gene: Gene that was
 targeted by qPCR (GltA or RpoB) jour and jour\_num: For nymphs, day after
 bloodmeal at which the tick was collected for DNA extraction. groupe\_Day:
 Compilation of antibiotic group and day group in the same variable.
 num\_tique: For adults, individual number associated with the tick.
 experimenter: Experimenter which performed the qPCR (experimenter1 or
 experimenter2) Ct\_GltA and Ct\_RpoB: cycle threshold at which the qPCR
 turned positive for GltA or RpoB gene Ct\_Actin: cycle threshold at which
 the qPCR turned positive for actin gene Ratio\_Ct\_GltA\_Ct\_Actin and
 Ratio\_Ct\_RpoB\_Ct\_Actin: Ratio of GltA or RpoB threshold / Actin
 threshold to normalize for the tick size. Concentration\_GltA and
 Concentration\_RpoB: Conversion of GltA or RpoB threshold into GltA or
 RpoB concentration using standard curves performed in the same qPCR.
 Concentration\_Actin: Conversion of actin threshold into actin
 concentration using standard curves performed in the same qPCR.
 Ratio\_des\_concentrations: Ratio of GltA or RpoB concentration / Actin
 concentration to normalize for the tick size. ConversionLog: Log
 conversion of the ratio of concentration for statistical analyses:
 =log(1+ratio) ## ReproductionTraits - summary The 15 datasheets made
 available here result from two campaigns of data collection and have been
 adjusted to be used into the R script
 script\_analysis\_of\_tick\_reproduction.R available in the same archive.
 This script can be launched using R version 4.2.3 and R studio and by
 installing the packages associated with the library used in the script
 (see ### packages and library ###). All datasheets can be extracted in the
 same folder from the archive ReproductionTraits.zip and used directly in
 the script after adding the path of the folder in the script. Data are
 issued from the monitoring of life history traits of soft ticks
 Ornithodoros moubata (female adults) treated or not with antibiotics
 and/or B vitamins to eliminate their microbiota. After blood feeding with
 treatments, female ticks were monitored during 2 periods of 50 days. We
 registered survival and traits associated with reproduction: laying
 success, laying date, number of eggs laid (standardized with the female
 tick weight as big ticks tend to lay more eggs), egg mass, date of
 hatching, hatching rate, date of molting into first nymphal stage, and
 molting rate. ## ReproductionTraits - Description of the Data and file
 structure Data were collected over two successive periods of 50 days
 following blood meal of the ticks. The file names indicate which period X
 is concerned: first 50-day period (1), second 50-day period (2) or both
 (1and2). For instance, Bloodmeal\_1\_complete contains all the data for
 the first period of 50 days. Here is the detail of how the information
 were collected/registered/transformed for each variable: “num\_tique“:
 Individual number associated with each tick (from F01 to F132) for
 randomization “groupe\_antibiotique“: Antibiotic treatment group of the
 tick (Rifampicin R, Gentamycin G or Control T). This information is added
 in the dataset after the collection of the data is over.
 “groupe\_vitamines“: Vitamin supplementation group of the tick (Vitamin
 Vit or NoVitamin NoVit) . This information is added in the dataset after
 the collection of the data is over. “groupe\_abvit“: Antibiotic treatment
 group and Vitamin supplementation group in the same variable. This
 information is added in the dataset after the collection of the data is
 over. “poids\_avant\_gorgement“: Weight of the tick the day before
 bloodmeal in mg. “poids\_apres\_gorgement“: Weight of the tick just after
 bloodmeal in mg. “poids\_apres\_ponte”: Weight at the end of the 50-day
 period. “poids\_apres\_gorgement\_deux”: Weight of the tick just after
 second bloodmeal after the first 50-day period in mg.
 “poids\_fin\_de\_manip”: Weight of the tick at the end of the full
 experiment. “date\_de\_la\_mort“: Date at which the tick died. (Empty if
 it didn’t die) “succes\_gorgement“: If the tick fed properly during
 bloodmeal, i.e. gained at least 50mg of weight (1 if the tick fed
 succesfully, 0 if it did not) “survie”: Survival (1 if the tick survived,
 0 if it died during the 50-day period) “succes\_ponte”: Success of laying
 eggs (1 if the tick laid eggs successfully during the 50-day period, 0 if
 it did not) “date\_de\_ponte“: Dates at which the tick laid eggs. The
 first date corresponds to the first time the tick laid eggs. The following
 dates correspond to all the dates which for eggs were laid by the tick.
 “jours\_avant\_ponte“: Number of days between the date of the bloodmeal
 and the date of laying eggs. In days. “nombre\_d\_oeufs“: Number of eggs
 laid in a patch of eggs laid the same day. “poids\_oeufs“: Weight of a
 patch of eggs laid the same day. “poids\_total\_oeufs“: Weight of all eggs
 laid by a same tick. “nombre\_total\_d\_oeufs\_avant\_exclusion“: Total
 number of eggs that were laid by the tick during the 50-day period. Some
 eggs were collected for qPCR analyses and were then excluded from the rest
 of the study (hatching rate etc…). This is the number of eggs before
 exclusion. “nombre\_total\_d\_oeufs\_apres\_exclusion“: Total number of
 eggs that were laid by the tick during the 50-day period. Some eggs were
 collected for qPCR analyses and were then excluded from the rest of the
 study (hatching rate etc…). This is the number of eggs after exclusion.
 “nombre\_total\_d\_oeufs\_avant\_exclusion\_div\_par\_poids“: Total number
 of eggs that were laid by the tick during the 50-day period divided by the
 weight of the tick mother (“poids\_apres\_gorgement“). Some eggs were
 collected for qPCR analyses and were then excluded from the rest of the
 study (hatching rate etc…). This is the number of eggs before exclusion
 divided by the tick weight. “poids\_moyen\_oeuf“: Average weight of 1 egg
 for a tick (“poids\_total\_oeufs“ divided by
 “nombre\_total\_d\_oeufs\_avant\_exclusion“). “date\_eclosion”: Date at
 which the eggs hatched. They can be several dates for a patch of eggs laid
 the same day if they didn’t hatch the same day.
 “temps\_incubation\_oeufs“: Number of days between the date of hatching
 and the date of laying eggs. In days. “nombre\_de\_larves\_ecloses“:
 Number of eggs from a patch of eggs that hatched that day.
 “oeufs\_non\_eclos“: Number of eggs from a patch of eggs that did not
 hatch at all. “nb\_total\_de\_larves“: Total number of larvae that hatched
 successfully by tick. “taux\_eclosion“: Hatching rate
 (“nombre\_de\_larves\_ecloses“ divided by
 “nombre\_total\_d\_oeufs\_apres\_exclusion“) “date\_de\_mue“: Date at
 which the larvae molted. They can be several dates for a patch of larvae
 hatched the same day if they didn’t molt the same day. “nb\_de\_mues“:
 Number of larvae that molted that day for each patch of larvae.
 “nb\_total\_de\_mues“: Total number of larvae that molted succesfully for
 a tick. “mortalite\_larves“: Number of larvae which died.
 “temps\_pour\_mue“: Number of days between the date of hatching and the
 date of molting. In days. “Survie“: Percentage of ticks that survived
 during the 50-day period, for a treatment condition. “erreur\_survie“: 95%
 confidence error on “Survie“ “ponte“: Percentage of ticks that laid at
 least one egg during the 50-day period, for a treatment condition.
 “erreur\_ponte“: 95% confidence error on “ponte“ “erreur\_survie\_old“:
 95% confidence error for “Survie“ to add to graph without going under 0%
 “erreur\_ponte\_old“: 95% confidence error on “ponte“ to add to graph
 without going under 0% “erreur\_survie\_plus“: 95% confidence error for
 “Survie“ to add to graph without going over 100% “erreur\_ponte\_plus“:
 95% confidence error on “ponte“ to add to graph without going over 100%
 Empty cells: Empty cells were not filled with "null," or
 "n/a," due to interference with the scripts. Empty cells occur
 either after an adult tick died before laying eggs, a group of eggs
 didn't hatch, or a group of larvae didn't molt, which is taken
 into account by the following variables: “survie”, “taux\_eclosion“, and
 “nb\_total\_de\_mues“ In these cases, all the columns which are no longer
 relevant contain empty cells (example: for a tick which died before laying
 eggs, dates at which the tick laid eggs “date\_de\_ponte” is an empty
 cell). ## Sharing/access Information Authors: Florian Taraveau
 (florian.taraveau@cirad.fr or f.taraveau@gmail.com) with the help of
 Helene Jourdan (helene.jourdan@cirad.fr) Affiliation of the authors: UMR
 ASTRE, CIRAD, INRAE, Campus de Baillarguet, F-34398 Montpellier, France
 Feel free to contact the authors for any questions on this dataset. DOI of
 the preprint which used these data:
 https://doi.org/10.1101/2023.05.09.539061