Female Saliva Research Articles

Disposable and flexible screen-printed-carbon electrode modified with Au/poly-beta-cyclodextrin as electrochemical platform for estriol detection

In this work, a screen-printed carbon electrode in polyethylene (SPPE) modified with Au particles and poly-beta-cyclodextrin polymeric film (SPPE/Au/p-beta-CD) was develop as an electrochemical platform to detect estriol (E3) in environmental and biological samples. The stability, reproducibility and sensitivity of the electrode were confirmed via physicochemical and electrochemical characterization. Electrochemical tests demonstrated an increase in the SPPE electroactive area when 70% graphite ink was applied, and transmission electron microscopy revealed an increase in surface complexity compared with that of the unmodified electrode. The electrochemical impedance spectroscopy (EIS) to SPPE/Au/p-beta-CD data indicated a decrease and increase in the Rct (0.096 kΩ cm2) and constant phase element (CPE) (28.34 µF cm−2) values, respectively. The analytical curve was plotted using concentrations ranging from 2.0 to 48.5 µmol/L and the best results were obtained at pH 4.0; a limit of detection (LD) of 1.88 µmol/L was obtained. During a study of interferents, the platform exhibited high selectivity for E3. The SPPE/Au/p-beta-CD exhibited recovery percentages of E3 between 100.32–110.17% in river water, and female urine, saliva, and sweat. The SPPE/Au/p-beta-CD as electrochemical platform proved to be a reproducible, selective, and low-cost sensor for the detection of E3 in different complex matrices. The modification of SPPE with Au and the beta-CD polymer made the electrode more sensitive and the method used is viable for real application.

Novel biomarkers in the saliva of healthy young males and females in a randomized crossover study on sedentary time: An exploratory analysis.

Several known biomarkers have been used to understand the physiological responses of humans to various short and long-term interventions such as exercise or dietary interventions. However, little exploratory work has been conducted to identify novel biomarkers in human saliva that could enable non-invasive physiological research to understand acute responses to interventions such as reducing sedentary time. The purpose of this study was to identify novel biomarkers in the saliva (cytokines, growth factors and vascular factors) that respond to prolonged (4 hours) and interrupted sitting (4 hours of sitting interrupted by 3 minutes of walking at 60% of maximal heart rate every 27 minutes) in young, healthy males and females. We also sought to determine whether responsive biomarkers would differ by sex. Participants (n = 24, 21.2 ± 2.2 years, 50% female) completed a prolonged sitting (PS) session and an interrupted sitting (IS) session in random order. Individual saliva samples were pooled into a male sample and a female sample to identify responsive biomarkers using a human cytokine antibody membrane array (42 targets). Several novel biomarkers were responsive in both sexes (e.g., IL-8, Angiogenin, VEGF, and EGF), in females only (e.g., TNF-α and IL-13), and in males only (e.g., IL-3, RANTES, and IL-12p40/p70). Importantly, several biomarkers appear to be responsive to the 4-hour prolonged and interrupted sitting sessions (e.g., TNF-α, IL-8, IL-3, RANTES, EGF, Angiogenin, and VEGF). This work highlights new directions for researchers aiming to investigate the effect of short-term or acute interventions on different physiological pathways using non-invasive methods. Our work clearly indicates that human saliva samples can provide a wealth of insight into physiological responses, and that a number of biomarkers can be used to understand changes induced by acute interventions such as interrupting prolonged sitting.

Changes in saliva protein profile throughout Rhipicephalus microplus blood feeding

BackgroundWhen feeding on a vertebrate host, ticks secrete saliva, which is a complex mixture of proteins, lipids, and other molecules. Tick saliva assists the vector in modulating host hemostasis, immunity, and tissue repair mechanisms. While helping the vector to feed, its saliva modifies the site where pathogens are inoculated and often facilitates the infection process. The objective of this study is to uncover the variation in protein composition of Rhipicephalus microplus saliva during blood feeding.MethodsTicks were fed on calves, and adult females were collected, weighed, and divided in nine weight groups, representing the slow and rapid feeding phases of blood feeding. Tick saliva was collected, and mass spectrometry analyses were used to identify differentially secreted proteins. Bioinformatic tools were employed to predict the structural and functional features of the salivary proteins. Reciprocal best hit analyses were used to identify conserved families of salivary proteins secreted by other tick species.ResultsChanges in the protein secretion profiles of R. microplus adult female saliva during the blood feeding were observed, characterizing the phenomenon known as “sialome switching.” This observation validates the idea that the switch in protein expression may serve as a mechanism for evading host responses against tick feeding. Cattle tick saliva is predominantly rich in heme-binding proteins, secreted conserved proteins, lipocalins, and protease inhibitors, many of which are conserved and present in the saliva of other tick species. Additionally, another remarkable observation was the identification of host-derived proteins as a component of tick saliva.ConclusionsOverall, this study brings new insights to understanding the dynamics of the proteomic profile of tick saliva, which is an important component of tick feeding biology. The results presented here, along with the disclosed sequences, contribute to our understanding of tick feeding biology and might aid in the identification of new targets for the development of novel anti-tick methods.Graphical

Salivary protein homology between humans and dogs: Mass spectrometry-based proteomics analysis

ObjectiveThis benchmark study aimed to investigate sex-related differences based on the identification and characterization of the salivary proteome of healthy male and female dogs using mass spectrometry (MS) technique and a homology-driven approach to analyze salivary proteins in both human and dog species utilizing protein sequence alignment technique. MethodsUnstimulated whole saliva was collected from 10 healthy Beagles. After processing the samples and determining the total protein content, in-solution protein digestion was performed involving denaturation, reduction of disulfide bonds, alkylation, and removal of interfering compounds. Samples were analyzed using LC-ESI-MS/MS. ResultsLC-ESI-MS/MS analysis identified 327 and 341 unique proteins in male and female dog saliva, respectively, of which 318 (97.25 %) in male dogs and 326 (95.60 %) in female dogs were characterized. Abundant shared proteins included albumin, BPI fold-containing family A member 2, and VWFD domain-containing protein. A notable uncharacterized protein, VWFD domain-containing protein, was among the most abundant in both sexes. Comparative analysis of 69 abundant shared proteins indicated an upregulation of CES5A, EFHD, GC, IGHM, LOC100653049, KRT10, LCP1, PGD, TPI1 in male dogs, while LOC100855593 was upregulated in female dogs. In total, 84 % (n = 229/274) and 86 % (n = 235/275) salivary proteins identified in male and female dogs, respectively, were homologous to human proteins, with an overall homology of 86 % (n = 364/423), including 15 with 100 % homology. ConclusionThe study revealed clear differences in the salivary proteomics profile of healthy male and female dogs. However, most of the salivary proteins in both male and female dogs showed homology with human salivary proteins. Clinical relevanceThe identification of unique salivary proteome profiles in male and female dogs, coupled with substantial homology to human proteins, provides promising biomarkers for health assessment, highlighting its clinical significance for diagnostics and therapeutic exploration not only in veterinary and human dentistry, but across mammalian species.

Performance evaluation of fully automated cobas® 6800 CMV PCR for the detection and quantification of cytomegalovirus DNA in neonatal urine and saliva, and adult urine, saliva, and vaginal secretion.

Laboratory testing for cytomegalovirus (CMV) in bodily fluids is essential to manage congenital and prenatal CMV infection. The rapid and fully automated cobas® CMV PCR is approved only for the testing of plasma in transplant patients. To evaluate the performance of the cobas® CMV to detect and quantify CMV DNA in neonatal and adult female urine, saliva, and vaginal secretion, the limit of detection (LoD), limit of quantification (LoQ), imprecision, linearity, PCR efficiency, bias, analytical specificity, cross-reactivity, and cross-contamination of the cobas® CMV for urine, saliva, and vaginal secretion was determined. The performance of the assay was evaluated prospectively with two laboratory-developed PCR assays using neonatal and adult urine, saliva swabs, and vaginal swabs. The LoD and LoQ were 31 and 100 IU/mL, respectively, for urine, and 81 and 100 IU/mL, respectively, for vaginal secretion. The LoD and LoQ for saliva were the same (200 IU/mL). The cobas® CMV was precise (coefficient of variation ≤10%), linear (R2 ≥ 0.995), and efficient (1.07 and 1.09) between 100 and 250,000 IU/mL for the sample types. The bias and analytical specificity was <±0.30 log10 IU/mL and 100%, respectively. Cross-reactivity with non-CMV pathogens was not detected. Cross-contamination rate was 0.28%. The diagnostic accuracy, sensitivity, and specificity of the cobas® CMV for neonatal urine and saliva were ≥95.0%, ≥93.3%, and ≥90.4%, respectively. The overall percent agreement for adult urine, saliva, and vaginal secretion was 86.6%, 94.5%, and 89.4%, respectively. Taken together, the cobas® CMV demonstrated acceptable analytical and diagnostic performance, and is suitable for routine diagnostic laboratory investigation of CMV infection in neonates and adults.

Evaluation of Interleukin-1β and Tumor Necrosis Factor Alpha Concentration in a Sample of Endodontic Patients in Hillah City, Iraq

Abstract Background: Endodontic illness, one of the most common chronic infectious diseases in the world, is brought on by an infected and inflamed dental pulp, which results in bone destruction around the tooth root, excruciating pain, and tooth loss. Objectives: This study aimed to determine levels of pro-inflammatory (interleukin-1β [IL-1β] and tumor necrosis factor alpha [TNF-α]) by enzyme-linked immune sorbent assay in the saliva of patients with endodontic infection against the gender and age groups. Materials and Methods: From November 2021 to March 2022, 60 patients (n = 60) were referred to specialized dentistry centers and private clinics in Hillah City, Iraq, with 32 (53.33%) females and 28 (46.67%) males, and 20 (n = 20) healthy adults serving as study controls, all of whom were between the ages of 20 and 50. Results: There were no major differences between girls and males in both groups, according to the findings of this study. In comparison to the control group, however, patients showed higher significant differences in measuring (IL-1β) and (TNF-α) concentrations (180.8), (119.44) (P ≤ 0.05), respectively. Furthermore, the amounts of IL-1β and TNF-α in female saliva were substantially greater than in male saliva. Conclusion: Pro-inflammatory IL-1β and TNF-α were shown to be higher in patients with primary endodontic disease than in normal subjects.

Gustatory polymorphism mediates a new adaptive courtship strategy.

Human-imposed selection can lead to adaptive changes in sensory traits. However, rapid evolution of the sensory system can interfere with other behaviours, and animals must overcome such sensory conflicts. In response to intense selection by insecticide baits that contain glucose, German cockroaches evolved glucose-aversion (GA), which confers behavioural resistance against baits. During courtship the male offers the female a nuptial gift that contains maltose, which expediates copulation. However, the female's saliva rapidly hydrolyses maltose into glucose, which causes GA females to dismount the courting male, thus reducing their mating success. Comparative analysis revealed two adaptive traits in GA males. They produce more maltotriose, which is more resilient to salivary glucosidases, and they initiate copulation faster than wild-type males, before GA females interrupt their nuptial feeding and dismount the male. Recombinant lines of the two strains showed that the two emergent traits of GA males were not genetically associated with the GA trait. Results suggest that the two courtship traits emerged in response to the altered sexual behaviour of GA females and independently of the male's GA trait. Although rapid adaptive evolution generates sexual mismatches that lower fitness, compensatory behavioural evolution can correct these sensory discrepancies.

Changes in Total Protein Level in Human Female Saliva during Ovulatory Functions and Metabolic Disorders

Saliva has been studied extensively as a potential diagnostic tool over the last decade due to its ease and non-invasive accessibility along with its abundance of biomarkers, such as genetic material and proteins.The activity of protein in saliva increased during ovulation. When we monitored salivary protein activity in 40 different women volunteers during various stages of reproduction like (prepubertal, parous, non-parous, menopausal and in the metabolic disorder state diabetic condition it has been observed that highly significant (p&lt;0.001) increase in parous ovulatory &amp; non parous ovulatory whereas, highly significant (p&lt;0.01)increase was observed in non-parous postovulatory and a highly significant (p&lt;0.001) decrease was observed in menopause and diabetic in comparison to prepubertal. A highly significant (p&lt;0.001) decrease was observed in menopause in comparison to parous preovulatory, ovulatory, post ovulatory &amp; non-parous ovulatory and postovulatory. A highly significant (p&lt;0.01) decrease was observed in diabetic in comparison to menopausal human female subjects. The result revealed that the total protein was considered as testing the saliva instead of blood isa non-invasive loom and it can be used as a biomarker for ovulation detection.

Comparative evaluation of periodontal health status of pre and postmenopausal females.

Menopause in females is a corporeal stage that gives rise to a number of adaptive changes both at systemic and oral levels, prototypically occurring in the late fourth or fifth decade of life. Though physiological aging affects the oral tissue, the hormonal changes due to menopause also act as a major contributing factor in deteriorating the health of oral tissues. Thus, the main aim of our study was to compare the overall periodontal status and alkaline phosphatase levels in the saliva of females in their pre and postmenopausal ages. This study was conducted on 200 female subjects coming to the Department of Periodontology at Rajendra Institute of Medical Science for oral prophylaxis. The subjects were arbitrarily selected in the age group ranging from 15 to 70 years and were further divided based on the inclusion criteria. Group A included 100 subjects with age ranging from 15 to 45 years, and group B, 100 subjects with age 54 to 70 years. Signs of periodontitis including clinical attachment loss, furcation involvement, and probing depth and salivary alkaline levels were obtained, evaluated, compared, and analyzed. In group A 65% of patients had grade 0 and 28% had grade 1 of clinical attachment level. Similarly in group B, 44% of the total had grade 0 of clinical attachment loss, 38% had grade 1, and 18% were of grade 2. On evaluating grades of furcation involvement, around 45% of the total patients in group A were grade 1 (incipient, pocket formation), while in group B, 51% of the total patients were grade 1. At least 46% in group A and only 20% in group B had no signs of furcation involvement. Salivary alkaline phosphatase levels in pre and postmenopausal patients showed a significant difference between the two groups was obtained. The study concluded a major difference in the periodontal health of pre and postmenopausal women with many influencing factors apart from menopause itself.

Sex-biased proteomic response to tomato spotted wilt virus infection of the salivary glands of Frankliniella occidentalis, the western flower thrips

Successful transmission of tomato spotted wilt virus (TSWV) by Frankliniella occidentalis requires robust infection of the salivary glands (SGs) and virus delivery to plants during salivation. Feeding behavior and transmission efficiency are sexually-dimorphic traits of this thrips vector species. Proteins secreted from male and female SG tissues, and the effect of TSWV infection on the thrips SG proteome are unknown. To begin to discern thrips factors that facilitate virus infection of SGs and transmission by F. occidentalis, we used gel- and label-free quantitative and qualitative proteomics to address two hypotheses: (i) TSWV infection modifies the composition and/or abundance of SG-expressed proteins in adults; and (ii) TSWV has a differential effect on the male and female SG proteome and secreted saliva. Our study revealed a sex-biased SG proteome for F. occidentalis, and TSWV infection modulated the SG proteome in a sex-dependent manner as evident by the number, differential abundance, identities and generalized roles of the proteins. Male SGs exhibited a larger proteomic response to the virus than female SGs. Intracellular processes modulated by TSWV in males indicated perturbation of SG cytoskeletal networks and cell-cell interactions, i.e., basement membrane (BM) and extracellular matrix (ECM) proteins, and subcellular processes consistent with a metabolic slow-down under infection. Several differentially-abundant proteins in infected male SGs play critical roles in viral life cycles of other host-virus pathosystems. In females, TSWV modulated processes consistent with tissue integrity and active translational and transcriptional regulation. A core set of proteins known for their roles in plant cell-wall degradation and protein metabolism were identified in saliva of both sexes, regardless of virus infection status. Saliva proteins secreted by TSWV-infected adults indicated energy generation, consumption and protein turnover, with an enrichment of cytoskeletal/BM/ECM proteins and tricarboxylic acid cycle proteins in male and female saliva, respectively. The nonstructural TSWV protein NSs - a multifunctional viral effector protein reported to target plant defenses against TSWV and thrips - was identified in female saliva. This study represents the first description of the SG proteome and secretome of a thysanopteran and provides many candidate proteins to further unravel the complex interplay between the virus, insect vector, and plant host.

A deeper insight into the sialome of male and female Ochlerotatus triseriatus mosquitoes

Over the last 20 years, advancements in sequencing technologies have highlighted the unique composition of the salivary glands of blood-feeding arthropods. Further biochemical and structural data demonstrated that salivary proteins can disrupt host hemostasis, inflammation and immunity, which favors pathogen transmission. Previously, a Sanger-based sialome of adult Ochlerotatus triseriatus female salivary glands was published based on 731 expressed sequence tag (ESTs). Here, we revisited O. triseriatus salivary gland contents using an Illumina-based sequencing approach of both male and female tissues. In the current data set, we report 10,317 DNA coding sequences classified into several functional classes. The translated transcripts also served as a reference database for proteomic analysis of O. triseriatus female saliva, in which unique peptides from 101 proteins were found. Finally, comparison of male and female libraries allowed for the identification of female-enriched transcripts that are potentially related to blood acquisition and virus transmission.

A proteomics informed by transcriptomics insight into the proteome of Ornithodoros erraticus adult tick saliva

BackgroundThe argasid tick Ornithodoros erraticus is the main vector of tick-borne human relapsing fever (TBRF) and African swine fever (ASF) in the Mediterranean Basin. The prevention and control of these diseases would greatly benefit from the elimination of O. erraticus populations, and anti-tick vaccines are envisaged as an effective and sustainable alternative to chemical acaricide usage for tick control. Ornithodoros erraticus saliva contains bioactive proteins that play essential functions in tick feeding and host defence modulation, which may contribute to host infection by tick-borne pathogens. Hence, these proteins could be candidate antigen targets for the development of vaccines aimed at the control and prevention of O. erraticus infestations and the diseases this tick transmits. The objective of the present work was to obtain and characterise the proteome of the saliva of O. erraticus adult ticks as a means to identify and select novel salivary antigen targets.MethodsA proteomics informed by transcriptomics (PIT) approach was applied to analyse samples of female and male saliva separately using the previously obtained O. erraticus sialotranscriptome as a reference database and two different mass spectrometry techniques, namely liquid chromatography–tandem mass spectrometry (LC–MS/MS) in data-dependent acquisition mode and sequential window acquisition of all theoretical fragment ion spectra MS (SWATH-MS).ResultsUp to 264 and 263 proteins were identified by LC–MS/MS in the saliva of O. erraticus female and male ticks, respectively, totalling 387 non-redundant proteins. Of these, 224 were further quantified by SWATH-MS in the saliva of both male and female ticks. Quantified proteins were classified into 23 functional categories and their abundance compared between sexes. Heme/iron-binding proteins, protease inhibitors, proteases, lipocalins and immune-related proteins were the categories most abundantly expressed in females, while glycolytic enzymes, protease inhibitors and lipocalins were the most abundantly expressed in males. Ninety-seven proteins were differentially expressed between the sexes, of which 37 and 60 were overexpressed in females and males, respectively.ConclusionsThe PIT approach demonstrated its usefulness for proteomics studies of O. erraticus, a non-model organism without genomic sequences available, allowing the publication of the first comprehensive proteome of the saliva of O. erraticus reported to date. These findings confirm important quantitative differences between sexes in the O. erraticus saliva proteome, unveil novel salivary proteins and functions at the tick–host feeding interface and improve our understanding of the physiology of feeding in O. erraticus ticks. The integration of O. erraticus sialoproteomic and sialotranscriptomic data will drive a more rational selection of salivary candidates as antigen targets for the development of vaccines aimed at the control of O. erraticus infestations and the diseases it transmits.Graphical

Proteomics informed by transcriptomics for a qualitative and quantitative analysis of the sialoproteome of adult Ornithodoros moubata ticks

BackgroundThe argasid tick Ornithodoros moubata is the main vector in mainland Africa of African swine fever virus and the spirochete Borrelia duttoni, which causes human relapsing fever. The elimination of populations of O. moubata would contribute to the prevention and control of these two serious diseases. Anti-tick vaccines are an eco-friendly and sustainable means of eliminating tick populations. Tick saliva forms part of the tick-host interface, and knowledge of its composition is key to the identification and selection of vaccine candidate antigens. The aim of the present work is to increase the body of data on the composition of the saliva proteome of adult O. moubata ticks, particularly of females, since in-depth knowledge of the O. moubata sialome will allow the identification and selection of novel salivary antigens as targets for tick vaccines.MethodsWe analysed samples of female and male saliva using two different mass spectrometry (MS) approaches: data-dependent acquisition liquid chromatography-tandem MS (LC–MS/MS) and sequential window acquisition of all theoretical fragment ion spectra–MS (SWATH-MS). To maximise the number of proteins identified, a proteomics informed by transcriptomics analysis was applied using the O. moubata salivary transcriptomic dataset previously obtained by RNA-Seq.ResultsSWATH-MS proved to be superior to LC–MS/MS for the study of female saliva, since it identified 61.2% more proteins than the latter, the reproducibility of results was enhanced with its use, and it provided a quantitative picture of salivary components. In total, we identified 299 non-redundant proteins in the saliva of O. moubata, and quantified the expression of 165 of these in both male and female saliva, among which 13 were significantly overexpressed in females and 40 in males. These results indicate important quantitative differences in the saliva proteome between the sexes.ConclusionsThis work expands our knowledge of the O. moubata sialome, particularly that of females, by increasing the number of identified novel salivary proteins, which have different functions at the tick–host feeding interface. This new knowledge taken together with information on the O. moubata sialotranscriptome will allow a more rational selection of salivary candidates as antigen targets for tick vaccine development.Graphical

Salivary spectrum evaluation with Raman’s Spectrometer

Background: Biofluids possess a lot of hidden informations. Along with serum, saliva beholds great potential which needs to be discovered. Certain physical properties of saliva need to be studied closely and thus can be put into use for various diagnostic purposes. Aims and Objectives: To find difference between male and female saliva through spectrometry. Design: 20 systemically sound subjects were selected and explained about the study. The subjects should not be consuming any medication or tobacco products. Materials and Methodology: Subjects were asked to collect unstimulated saliva in the disposable vials. The samples were then centrifuged with the help of speed vacuum concentrator. Then with the help of Gilson’s pipette some centrifuge sample were taken and normal saline was added and was left to dry. With the help of Raman’s Spectrometer 585nm the spectra was recorded and each sample were assessed 5 times. And then raw data was processed. Result and Conclusion: PCA (Principal Component Analysis), LDA (Linear Discriminant Analysis) and Average Spectra are derived. The LDA accuracy of our study was above 70% which is good for future research purposes as anything above 60% can be considered as important breakthrough.

Vector competence of Aedes aegypti from Havana, Cuba, for dengue virus type 1, chikungunya, and Zika viruses.

BackgroundLike many countries from the Americas, Cuba is threatened by Aedes aegypti-associated arboviruses such as dengue (DENV), Zika (ZIKV), and chikungunya (CHIKV) viruses. Curiously, when CHIKV was actively circulating in the region in 2013–2014, no autochthonous transmission of this virus was detected in Havana, Cuba, despite the importation of chikungunya cases into this city. To investigate if the transmission ability of local mosquito populations could explain this epidemiological scenario, we evaluated for the first time the vector competence of two Ae. aegypti populations (Pasteur and Párraga) collected from Havana for dengue virus type 1 (DENV-1), CHIKV, and ZIKV.Methodology/Principal findingsMosquito populations were fed separately using blood containing ZIKV, DENV-1, or CHIKV. Infection, dissemination, and transmission rates, were estimated at 3 (exclusively for CHIKV), 7, and 14 days post exposure (dpe) for each Ae. aegypti population-virus combination. Both mosquito populations were susceptible to DENV-1 and ZIKV, with viral infection and dissemination rates ranging from 24–97% and 6–67% respectively. In addition, CHIKV disseminated in both populations and was subsequently transmitted. Transmission rates were low (<30%) regardless of the mosquito population/virus combination and no ZIKV was detected in saliva of females from the Pasteur population at any dpe.Conclusions/SignificanceOur study demonstrated the ability of Ae. aegypti from Cuba to transmit DENV, ZIKV, and CHIKV. These results, along with the widespread distribution and high abundance of this species in the urban settings throughout the island, highlight the importance of Ae. aegypti control and arbovirus surveillance to prevent future outbreaks.

Assessing the role of two populations of Aedes japonicus japonicus for Zika virus transmission under a constant and a fluctuating temperature regime

BackgroundSince the huge epidemic of Zika virus (ZIKV) in Brazil in 2015, questions were raised to understand which mosquito species could transmit the virus. Aedes aegypti has been described as the main vector. However, other Aedes species (e.g. Ae. albopictus and Ae. japonicus) proven to be competent for other flaviviruses (e.g. West Nile, dengue and yellow fever), have been described as potential vectors for ZIKV under laboratory conditions. One of these, the Asian bush mosquito, Ae. japonicus, is widely distributed with high abundances in central-western Europe. In the present study, infection, dissemination and transmission rates of ZIKV (Dak84 strain) in two populations of Ae. japonicus from Switzerland (Zürich) and France (Steinbach, Haut-Rhin) were investigated under constant (27 °C) and fluctuating (14–27 °C, mean 23 °C) temperature regimes.ResultsThe two populations were each able to transmit ZIKV under both temperature regimes. Infectious virus particles were detected in the saliva of females from both populations, regardless of the incubation temperature regime, from 7 days post-exposure to infectious rabbit blood. The highest amount of plaque forming units (PFU) (400/ml) were recorded 14 days post-oral infection in the Swiss population incubated at a constant temperature. No difference in terms of infection, dissemination and transmission rate were found between mosquito populations. Temperature had no effect on infection rate but the fluctuating temperature regime resulted in higher dissemination rates compared to constant temperature, regardless of the population. Finally, transmission efficiency ranged between 7–23% and 7–10% for the constant temperature and 0–10% and 3–27% under fluctuating temperatures for the Swiss and the French populations, respectively.ConclusionsTo the best of our knowledge, this is the first study confirming vector competence for ZIKV of Ae. japonicus originating from Switzerland and France at realistic summer temperatures under laboratory conditions. Considering the continuous spread of this species in the northern part of Europe and its adaptation at cooler temperatures, preventative control measures should be adopted to prevent possible ZIKV epidemics.

Gender-associated differences in oral microbiota and salivary biochemical parameters in response to feeding.

Saliva plays a key role in food absorption and digestion mainly due to both its enzymes and microbiota. The main objective of this study was to compare the oral microbiota and salivary parameters between men and women in response to feeding. To answer this question, we set up a pilot study on 10 male and 10 female subjects to examine the role of saliva in glycaemia physiology. Biological parameters and the microbiotal composition of saliva were analyzed in fasted and fed states. The results show that the level of blood glucose was not different between men and women in the fasted state (88.00mg/dL ± 6.38 vs 87.00mg/dL ±8.07, p= 0.9149) or in the fed state (102.44mg/dL ± 14.03 vs 116.9mg/dL ± 25, p= 0.1362). Free fatty acids (FFA 0.15mmol/L ± 0.15 vs 0.07mmol/L ± 0.07, p = 0,0078), cholesterol (0.53mmol/L ± 0.30 vs 0.15mmol/L ± 0.14, p< 0.0001), and total saliva proteins (13.2g/L ± 4.31 vs 9.02g/L ± 6.98, p= 0.0168) were decreased after feeding, as well as the saliva lipase (27.89U/L ± 25.7 vs 12.28U/L ± 4.85, p= 0.0126). A very significant increase in the relative abundance of Streptococcaceae (24.56 ± 9.32 vs 13.53 ± 7.47, p= 0.00055) and a decrease in Prevotellaceae (34.45 ± 9.30 vs 17.43 ± 9.03, p=0.00055) were observed in the fed condition. When investigating gender-related differences in the fasted state, men showed higher levels of cholesterol (0.71mmol/L ± 0.26 vs 0.40mmol/L ± 0.27, p= 0.0329), FFA (0.25mmol/L ± 0.18 vs 0.08mmol/L ± 0.06, p= 0.0049), and triglycerides (0.24mmol/L ±0.15 vs 0.09mmol/L ± 0.04, p= 0.006) than women. Finally, differences could be observed in saliva microbiota between men and women in the fasted condition but even more in the fed condition, where Porphyromonas and Capnocytophaga were overrepresented in the male salivary samples compared with female saliva. Thus, biological parameters and microbiota in saliva could be the signatures of the feeding conditions and sex gender status.

Rhipicephalus microplus serpins interfere with host immune responses by specifically modulating mast cells and lymphocytes

Rhipicephalus microplus ticks feed on a bovine host for three weeks. At the attachment site, inflammatory and immune responses are triggered resulting in the recruitment of cells and production of a set of immunological mediators. To oppose the host’s immune responses, ticks inoculate bioactive salivary molecules capable of interfering with these defense mechanisms. Serpins are among the most frequent molecules present in tick saliva and have been shown to negatively affect the host’s anti-tick immunity. R. microplus has at least eighteen full-length serpins (RmS) and eleven are transcribed during blood feeding. Among them, RmS-3, RmS-6, and RmS-17 are present in the saliva of engorged females. Here, the effect of these serpins on the immune responses was evaluated in cells involved in innate/inflammatory (mast cells and macrophages) and adaptive (T cells) immunity. RmS-3 modulated mast cells due to its inhibitory activity on peritoneal rat chymase and on vascular permeability in acute inflammation. In addition, both RmS-6 and RmS-17 inhibited vascular permeability. Of the three serpins studied, neither affected activation nor inflammatory cytokine production by murine macrophages. On the other hand, RmS-3 and RmS-17 presented an inhibitory effect on the metabolic activity of lymphocytes, with the latter being the most potent, while RmS-6 had no effect on it. This activity was associated with a decrease in lymphocyte proliferation, but not with induction of cell death. The present study highlights the powerful modulatory role of tick salivary serpins in the host’s immune system and inspire the discovery of targets for the treatment of inflammatory/immune disorders.

XIST and RPS4Y1 long non-coding RNA transcriptome as sex biomarkers in different body fluids

Background and objectivesSex determination of an individual based on biological fluid evidence is a critical issue in forensic science. RNA has proved valuable in the identification of body fluids and the estimation of stain age. However, it still could not provide sufficient information about their donor. This study aimed to evaluate the potential use of the long non-coding XIST and RPS4Y1 markers in the identification of sex from different body fluids.MethodsSaliva, semen, and peripheral and menstrual blood samples were obtained from apparently healthy volunteers. The expression of XIST and RPS4Y1 was assessed in these samples using real-time RT-PCR.ResultsXIST was detected in female saliva and peripheral and menstrual blood, while RPS4Y1 was detected in male blood and semen.ConclusionXIST and RPS4Y1 could be sex differentiation biomarkers in various body fluids.

Immunomodulatory Effects of Amblyomma variegatum Saliva on Bovine Cells: Characterization of Cellular Responses and Identification of Molecular Determinants.

The tropical bont tick, Amblyomma variegatum, is a tick species of veterinary importance and is considered as one of major pest of ruminants in Africa and in the Caribbean. It causes direct skin lesions, transmits heartwater, and reactivates bovine dermatophilosis. Tick saliva is reported to affect overall host responses through immunomodulatory and anti-inflammatory molecules, among other bioactive molecules. The general objective of this study was to better understand the role of saliva in interaction between the Amblyomma tick and the host using cellular biology approaches and proteomics, and to discuss its impact on disease transmission and/or activation. We first focused on the immuno-modulating effects of semi-fed A. variegatum female saliva on bovine peripheral blood mononuclear cells (PBMC) and monocyte-derived macrophages in vitro. We analyzed its immuno-suppressive properties by measuring the effect of saliva on PBMC proliferation, and observed a significant decrease in ConA-stimulated PBMC lymphoproliferation. We then studied the effect of saliva on bovine macrophages using flow cytometry to analyze the expression of MHC-II and co-stimulation molecules (CD40, CD80, and CD86) and by measuring the production of nitric oxide (NO) and pro- or anti-inflammatory cytokines. We observed a significant decrease in the expression of MHC-II, CD40, and CD80 molecules, associated with decreased levels of IL-12-p40 and TNF-α and increased level of IL-10, which could explain the saliva-induced modulation of NO. To elucidate these immunomodulatory effects, crude saliva proteins were analyzed using proteomics with an Orbitrap Elite mass spectrometer. Among the 336 proteins identified in A. variegatum saliva, we evidenced bioactive molecules exhibiting anti-inflammatory, immuno-modulatory, and anti-oxidant properties (e.g., serpins, phospholipases A2, heme lipoprotein). We also characterized an intriguing ubiquitination complex that could be involved in saliva-induced immune modulation of the host. We propose a model for the interaction between A. variegatum saliva and host immune cells that could have an effect during tick feeding by favoring pathogen dissemination or activation by reducing the efficiency of host immune response to the corresponding tick-borne diseases.