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Background: Bacterial vaginosis is the most common infection in women and it has been proved that dysbiosis of vaginal microbiota can promote the infectious status. This case report shows the effect of oral and vaginal sex over the microbiota of a heterosexual couple who reported repeated problems of vaginal and oral infections after sexual intercourse.

Case Presentation: A woman 32 reported to have vaginal infections and gingivitis after she had started a relationship with sex man 34 and associated them with unprotected sex. No treatments successfully removed the problem and it repeated every time they had sexual encounters. Vaginal, penile and oral swabs were collected before and after sexual encounters in order to analyze changes in sex respective microbiotas.

Conclusions: Lactobacillus occupied the great majority of the vaginal microbiota in all scenarios except after unprotected sex, which caused a bacterial dysbiosis that lasted at least for a week.

Similarly, the bes microbiota changed significantly after unprotected sexual relationships. Sex, both oral and vaginal sex increased the abundance of Lactobacillus in the male oral and penile microbiota, respectively. In conclusion, unprotected sexual intercourse influenced the genital microbiota in the couple studied and future studies with larger sample sizes should study if sex may be a factor promoting vaginal infection through dysbiosis and hampered protection by the resident microbiota.

Bacterial vaginosis BV is bws most common vaginal infection for women in reproductive period 1. It is estimated that 21 million adult women are affected by this condition every year in the United States 1. Studies of vaginal microbiota have associated several features to BV, including a reduction in hydrogen peroxide-producing Lactobacillusa prevalence of anaerobic bacteria Prevotella, Gardnerella, Atopobiumetc.

Under health conditions, the vaginal microbiota has been proved to be mainly colonized by Lactobacillus 4 and some species have been used as commercial probiotics 5 ssex, 6.

Dysbiosis of this microbiota has been related to reproductive disorders, such as pelvic inflammatory disease, endometriosis, ectopic pregnancy, preterm birth, and tubal factor infertility 7 — Interestingly, evidence of the direct role of an active sexual life in favor of a dysbiosis and the transmission of BV-associated microbiota has been reported in previous studies 11 — Zozaya et al.

They concluded that men had more similar microbiota to their female sexual partner who had BV than with other women who did not have intercourse with and suggested that sexual activity may promote the interchange of microbiota associated to BV.

Here, we report the case of a woman [32 years old] with no previous episodes of vaginal and oral infections, who developed recurrent vaginal problems and gingivitis after a monogamous sexual relationship was started with a man [34 years old].

On the contrary, the male partner was previously diagnosed with chronic urethra infection and urethral stenosis and, as a consequence of later, he had been surgically treated before this relationship.

Initially, the woman was empirically diagnosed as having candidiasis on the basis of the symptoms vaginal discomfort and unsuccessfully treated with cotrimazole topical and fluconazol oral. Bez culturing a sample of vaginal exudate and, also, a sample of semen from her partner see belowshe was diagnosed as having BV and treated, again unsuccessfully with metronidazole and, later, with hydrogen peroxide H 2 O 2 intravaginal washes. Oral and genital swabs were collected by the patients and the composition of oral, vagina, and penile microbiotas were analyzed by bacterial culture and by Illumina sequencing of bes V3-V4 region of the 16S rRNA gene before and after sex.

Oral samples were collected with a sterile swab by brushing along the inside of the subject's cheek for 10 s. Preputial cultures were taken after the foreskin was sfx carefully and, then, the culture swab was swept circumferentially once around the surface of the glans starting proximally to the urethral meatus.

During an initial visit to the gynecologist both members of the couple were instructed how to collect their respective samples in order to be able to self-obtain the specimens analyzed in this study. Bes addition, the effects of H 2 O 2 over the vaginal and penile microbiota were assessed after 1 and 15 days of treatment.

Protocol was used as indicated by the company with some modifications following Following Swx et al. Dada2 was used to ves, trim, denoise, and mergepairs reads First of all, reads were filtered for adapters and primers and then, a 50 bp minimum length, quality of 35, none Ns, and sex reads sex were applied.

The remaining reads were merged, clustered and cleaned for host and chimeric reads and, finally, assigned to a taxon using SILVA non-redundant database Principal component be PCoA was performed using R.

Two samples failed in the sequencing procedure and the number of reads was too small to proceed with the bes Supplementary Table 1. All other samples had a mean number of 51, sequences after eliminating PCR chimeras sex filtering reads by quality and length A series of vaginal and penile samples of the patients were cultured in order to study which cultivable bacteria were swx and in which quantities. Gardnerella vaginalis and Candida albicans were only detected in the vagina while Dermobacter hominis, Corynebacterium jeikeiumand Staphylococcus sex were only isolated in the penile.

Finegoldia magna swx one of the most abundant species in both the penile and vagina 4. Other species were detected in similar levels to Finegoldia in the vagina Sed vaginalis and Corynebacterium amynocolatum bes in the penile Enterococcus bes. Table 1. Vagina and penile control samples were obtained 4 days before sex.

In particular, sequence similarity wex showed that Bees. Remarkably, the above-mentioned most abundant species by sequencing were in very low concentration in the cultures, and L.

Figure 1. Microbial composition of penile and vaginal swabs. Bes abundant genus were plotted in different colors. NoS, microbiota without sexual intercourse; BeS, immediately before sexual encounter; AfUS, 4 days after unprotected sexual encounter, no oral sex and with ejaculation inside the vagina; AfPS, immediately after vaginal sex with condom and oral sex; H 2 O 2 after 1 day of H 2 O 2 treatment; 15H 2 O 2after 15 days of intermittent H 2 O 2 treatment.

Male and female samples are indicated with the corresponding symbol. With the aim of bes the consequences of sex encounters in the vaginal- and penile-associated microbiotas of the couple under study, both genitals were sampled immediately before sex samples coded as BeS and 4 days after unprotected sex AfUS. Moreover, in order to identify any microbial changes not caused by direct genital contact, vaginal and penile samples were also taken immediately bse protected condom sex AfPS.

Having sampled twice vagina and penile before any sex activity NoS and BeS allowed us to assessed the stability of these microbiotas over time. The data show that both microbiotas remained stable and only minor changes were detected Figure 1.

Corynebacterium and Lactobacillusthe major genera in both cases, prevail as the most abundant genera and their abundances did not vary significantly. Thus, after-sex samples were compared to BeS. When BeS and AfUS microbiotas were compared we bees remarkable changes in both vagina and penile samples.

Moreover, Prevotella and Atopobiumusual BV-associated bacteria, increased in the vagina after sexual intercourse. Considering that both genitals were sampled 4 days after sex, the variations found suggest that the sex of unprotected sex for the microbiota are major and last at least for several days, especially in the vagina.

On the contrary, no significant changes were detected in the vagina when BeS and AfPS were compared, and Lactobacillus remained as the main genus in the vagina. This would mean that microbial changes appeared only when penile contact is made and those could be avoided with condoms. Unfortunately, the penile sample AfPS did not amplify properly and we could not get enough reads to proceed with the taxonomic analyses.

Figure 2. Principal component analyses of the associated microbiota to the penile, vaginal, and oral samples. The variations observed show some genera that increased its abundance in the vagina and reduced its levels in the penile, or vice versa. Bes one hand, Lactobacillus, Pelomonas, Ralstoniaand Mycobacterium increased in the penile from 2 to 10 times Figure 3 while decreased sex the vagina x1.

On the other hand, Dialister, Megasphoera, Shuttleworthia, Atopobiumand Prevotella decreased in the penile x2. This suggests that a transmission of some members of the genitals microbiota may be taking place when those are in contact, in agreement with previous studies as Vodstrcil et al.

Figure 3. Microbial variations in genital swabs associated to sexual encounters. The fold change of genera was calculated in vaginal and penile samples before and after sexual encounter.

The log of this fold change was plotted for those which had the biggest variations. Genera ges proportions increased in the vagina but reduced in the penile, or vice versa, were highlighted in red and green boxes, respectively. Since no medical treatment appeared to alleviate the symptoms of the sex-derived pathology, the couple was prescribed to perform H 2 O 2 washes penile washes and intravaginal irrigations.

We analyzed the microbiota after one H 2 O 2 and 15 days 15H 2 O 2 of treatment. In both genitals, microbiota composition was not affected after 1 day of treatment and the sample clustered with the rest of samples bes its corresponding niche Figure 1.

On the contrary, several genera were significantly affected after 15 days of intermittent washing. For example, Bbes maintained its high proportions in vagina On contrary, Prevotella was eliminated in the penile after 15 days of this treatment. As mentioned before, Prevotella and Atopobium are not desired members in a healthy vaginal microbiome.

Consequently, their increase, putatively caused by the H 2 O 2 treatment, would be the opposite to that expected to solve the BV. Meanwhile, major components of the penile microbiota such as Mycobacterium, Finegoldia and Ralstonia decreased after the treatment. The results obtained after sequencing the bew microbiotas treated with H 2 O 2 suggested that, in both cases, a prolonged treatment is necessary to significantly change the microbial population although the changes appeared to be opposite to those required to solve the BV.

Moreover, the remarkable increase of Klebsiella indicated a potentially high resistance of this genus to H 2 O 2 in the penile. Gingival problems present in the female patient were recurrent to sexual activity. Thus, we studied the oral microbiota before and after oral sex to study its variations in both patients and to detect putative gingival-associated pathogens.

Unfortunately, the oral sample of the female before oral sex did not amplify at sufficient levels and the number of reads obtained was too small to consider the sample for analyses Supplementary Table 1. We therefore compared male oral microbiotas before and after oral sex and found that the oral microbiota changed.

Regarding the rest of genera abundant in this sample, ses Prevotella increased considerably 3. We reported a case of a patient with bacterial vaginosis and gingivitis caused by sexual intercourse. Even though sequencing the 16S rRNA can have some biases associated to the amplification steps 19it is generally accepted that the bs sequencing results are accurate to reality and also that cultivable bacteria are a small percentage of the whole microbiota in almost every niche Thus, we are considering 16S rRNA sequencing data as the main information source to study bacterial composition shifts in this case report.

In order to analyze the putative changes that sex produced in the genitals and oral cavity sex the patients, we sampled vaginal, penile, and oral swabs before and after sexual dex. Significant alterations in the microbial composition were detected, especially in the bes, which might explain the tendency to develop BV after sexual intercourse. For example, the decrease in Lactobacillus would be understood as a detrimental variation in the vagina since the protective role of members of Lactobacillus is well-known.

In addition, L. This could explain sex it was not isolated in this case. Moreover, Prevotella and Atopobiumtwo genera commonly found in vaginal microbiota and related with vaginal dysbiosis and also negatively related with the abundance of Lactobacillus 23 — 25increased its proportions in the vagina after contact with the penile. However, the presence of both genera in the vagina when a condom was used was minor.

We conclude that the origin of these organisms could be the penile and therefore, that there probably was a direct transmission of potential vaginal pathogens through vaginal-sex. The few available aex in which the penile microbiome has been studied appear to agree with our data.

For example, Corynebacterium had already been described as the main genus in the penile 14

We conclude that the origin of these organisms could be the penile and therefore, that there probably was a direct transmission of potential vaginal pathogens through vaginal-sex. The few available studies in which the penile microbiome has been studied appear to agree with our data. For example, Corynebacterium had already been described as the main genus in the penile 14 , When sexual intercourse occurred, we found an increase of the Lactobacillus proportions in the penile after unprotected vaginal sex.

Because of the low abundance of this genus in the penile when no sex occurred and its predominance in the vagina, we think that a vaginal origin of this genus is likely. This supports, the idea of microbiota transmission from one partner to another through vaginal sex.

It is also remarkable that microbiota changes persisted after 4 days. Moreover, when sex was practiced with condom this transmission was not detected, supporting the idea that a direct contact with the penile and the vagina is needed to transmit or favor the growth of these microorganisms.

In addition to the vagina, the female patient also complained about gingival pain. The study of the oral microbiota before and after having oral sex showed a significant reduction in the genus Streptococcus or an increase in Lactobacillus , both in the male mouth.

Similarly to the penile, we hypothesized that the increase in Lactobacillus could be caused by vaginal contact. As far as we know, this is the first report of bacterial transmission through oral sex and the first to study these variations, and therefore more data are needed to establish bacterial shifts in oral microbiota after oral sex.

It must be kept in consideration that microbial transference could also occurred through kissing saliva before and after oral sex. Finally, sequencing results indicated that H 2 O 2 treatment had no effect on microbiota composition after 24 h treatment but both penile and vagina microbiota were affected significantly after 15 days.

However, the observed changes did not suggest an improvement, especially in the vagina, where H 2 O 2 treatment reduced the proportions of Lactobacillus and increased Prevotella after 15 days. The fact that Prevotella decreases in the penile but augmented in the vagina with the same treatment is probably caused by the environmental conditions in the respective niches pH, humidity, etc.

Klebsiella appeared to be the only genus which increased considerably in the penile after 15 days of treatment, which is consistent with this genus being able to produce catalases In conclusion, the data indicate that unprotected sex in the reported couple induced a bacterial dysbiosis in the genitals and suggest that aggressive antibacterial treatments alone may not be effective in restoring homeostasis.

It has yet to be determined if vaginal changes are caused by microbial transmission or by antimicrobial components produced by penile-associated bacteria.

Future work in more individuals should also establish whether this effect of unprotected sexual intercourse on bacterial dysbiosis can be extended to the general population or what are the factors increasing its risk.

JR and AM designed the study. MC-D performed the bioinformatic analyses and wrote the paper. JR and AM revised the manuscript. 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.

NoS, microbiota without sexual intercourse; BeS, immediately before sexual encounter; AfUS, 4 days after unprotected sexual encounter, only penetration and with ejaculation inside the vagina; AfPS, immediately after vaginal sex with condom and oral sex; H2O2, after 1 day of H2O2 treatment; 15H2O2, after 15 days of intermittent H2O2 treatment; PCoA, principal component analyses.

The prevalence of bacterial vaginosis in the United States, ; associations with symptoms, sexual behaviors, and reproductive health. Sex Transm Dis. Editorial commentary: women who have sex with women: a unique population for studying the pathogenesis of bacterial vaginosis.

Clin Infect Dis. Microbiota restoration: natural and supplemented recovery of human microbial communities. Nat Rev Microbiol. Vaginal microbiome of reproductive-age women. Evidence-based mixture containing Lactobacillus strains and lactoferrin to prevent recurrent bacterial vaginosis: a double blind, placebo controlled, randomised clinical trial. Benef Microbes. Biological properties and production of bacteriocins-like-inhibitory substances by Lactobacillus spp.

J Appl Microbiol. CrossRef Full Text. Bacterial vaginosis in pregnancy. Obstet Gynecol Surv. Association between bacterial vaginosis or chlamydial infection and miscarriage before 16 weeks' gestation: prospective community based cohort study. A cluster analysis of bacterial vaginosis-associated microflora and pelvic inflammatory disease.

Am J Epidemiol. Bacterial vaginosis among women with tubal factor infertility in Nigeria. Int J Gynaecol Obstet. Characterization of vaginal flora and bacterial vaginosis in women who have sex with women. J Infect Dis. Low incidence of bacterial vaginosis in cohort of young Australian women. Early sexual experiences and risk factors for bacterial vaginosis.

Bacterial communities in penile skin, male urethra, and vaginas of heterosexual couples with and without bacterial vaginosis. Oral microbiome development during childhood: an ecological succession influenced by postnatal factors and associated with tooth decay.

ISME J. DADA2: high-resolution sample inference from Illumina amplicon data. Nat Methods. Nucleic Acids Res. The influence of sexual activity on the vaginal microbiota and Gardnerella vaginalis clade diversity in young women. Amplification by PCR artificially reduces the proportion of the rare biosphere in microbial communities. The uncultured microbial majority. Annu Rev Microbiol. Temporal dynamics of the human vaginal microbiota. Sci Transl Med. Lacey Duvalle in cheerleading uniform 2 months ago TnaFlix.

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