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The microbiota plays an important role in numerous physiological processes, pathogenesis, development, and metabolism in different animal species. In humans, several studies have demonstrated an association between the vaginal microbiota and fertility rates, and even success in assisted reproduction techniques. In the context of cattle reproduction, although few studies have addressed the microbiota in a healthy state (which is not associated with diseases that affect the reproductive tract of cows), changes in its composition also seem to influence fertility. This review aims to explain the importance of the reproductive microbiota in female bovines and what is available in the literature regarding its possible role in increasing fertility. What are the challenges involved in this process? Future perspectives on its use and manipulation as a selection or intervention tool. Will it be possible to one day extrapolate the findings to reality and apply them in the field? In short, understanding the role of the reproductive microbiota of female bovines can signal the prospect of increasing production, whether of milk or meat, from the same number of animals, as it can optimize reproductive efficiency and perhaps become an allied tool for the economic profitability and sustainability of livestock farming.
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El parto prematuro (PP) es la principal causa de morbilidad/mortalidad perinatal en el mundo. La infección intrauterina es el origen más frecuente del PP espontáneo (PPE) en un hospital público de Chile. Existe evidencia de que la infección bacteriana ascendente (IBA) produce la infección/inflamación intraamniótica, el PPE y los resultados adversos maternos y perinatales. Esta revisión narrativa incluye revisiones sistemáticas y estudios de cohorte o de caso-control sobre la microbiota y el perfil inmunológico existente en el tracto genital inferior (TGI) de la embarazada propensa a PPE por IBA. Existe consenso en que en la microbiota del TGI de esta gestante hay colonización vaginal con baja abundancia de lactobacilos y/o disminución de su calidad, por diferencias raciales y/o geográficas o genéticas y una desregulación de los mecanismos inmunológicos del TGI. Estas respuestas se presentan con mayor intensidad en pacientes con factores de riesgo del huésped, como diabetes, obesidad, estrés, ansiedad y depresión, originando infecciones recurrentes del TGI, responsables del PPE y de los resultados perinatales. El conocimiento del comportamiento de la microbiota y del sistema inmunitario en estos casos permitirá tener terapias eficaces para prevenir el PPE y la morbilidad/mortalidad neonatal por IBA.
Preterm delivery (PD) is the leading cause of perinatal morbidity/mortality in the world. Intrauterine infection is the most frequent origin of spontaneous PD (SPD) in a public hospital in Chile. There is evidence that vaginal ascending bacterial infection (ABI) causes intra-amniotic infection/inflammation, SPD, and adverse maternal and perinatal outcomes. This narrative review includes systematic reviews and cohort or case-control studies on the microbiota and immunological profile existing in the lower genital tract (LGT) of pregnant women prone to SPD due to ABI. There is consensus that in the LGT microbiota of this pregnant woman there is vaginal colonization with low abundance of Lactobacilli and/or decreased quality, due to racial and/or geographic, or genetic differences and dysregulation of immunological mechanisms of the LGT. These responses occur with greater intensity in patients with host risk factors, such as diabetes, obesity, stress, anxiety, and depression, causing recurrent LGT infections responsible for SPD and perinatal outcomes. Knowledge of the behavior of the microbiota and the immune system in these cases will allow effective therapies to prevent SPD and neonatal morbidity/mortality due to ABI.
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Humanos , Feminino , Gravidez , Complicações Infecciosas na Gravidez , Vagina/imunologia , Vagina/microbiologia , Trabalho de Parto PrematuroRESUMO
Background/Purpose: Polycystic ovary syndrome (PCOS) is an endocrine-metabolic disease most common in patients of childbearing age. This pathology is associated with clinical, metabolic, and reproductive complications. We evaluated the diversity of the vaginal microbiota (VM), the vaginal inflammatory reaction (VIR), the proinflammatory state, and the activation of polymorphonuclear neutrophils (PMN) with the production of neutrophil extracellular traps (NETs). Methods: Thirty-three patients who attended a consultation at the Hospital UTPL-Santa Inés, Loja, Ecuador, from May to August 2023 who were diagnosed with PCOS participated in this study. Blood samples, vaginal discharge, and a survey were obtained. Results: A high number of patients, 23/33 (69.7%), presented altered microbiota in clinical variables associated with PCOS phenotypes A and B, sexual partners (>2), and oligomenorrhoea. A significant statistical association was only observed for sexually transmitted infections at sampling (p = 0.023) and insulin (p = 0.002). All eight cases studied with VIR had PMN/NETotic activity. A high frequency of proinflammatory states was observed in all vaginal microbiota states. Conclusions: These results suggest that the PCOS could trigger a proinflammatory state in the vaginal epithelium independently of the state of the vaginal microbiota. Furthermore, the presence of NETs observed in the cases studied could decrease fertility in these PCOS patients.
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Cervical cancer ranks among the most prevalent cancers globally with high-risk human papillomaviruses implicated in nearly 99% of cases. However, hidden players such as changes in the microbiota are now being examined as potential markers in the progression of this disease. Researchers suggest that changes in the vaginal microbiota might correlate with cervical cancer. This review provides a comprehensive look at the microbiota changes linked with the advancement of cervical cancer. It also scrutinizes the databases from past studies on the microbiota during healthy and cancerous stages, drawing connections between prior findings concerning the role of the microbiota in the progression of cervical cancer. Preliminary findings identify Fusobacterium spp., Peptostreptococcus spp., Campylobacter spp., and Haemophilus spp., as potential biomarkers for cervical cancer progression. Alloscardovia spp., Eubacterium spp., and Mycoplasma spp. were identified as potential biomarkers for HPVs (+), while Methylobacterium spp. may be indicative of HPV (-). However, the study's limitations, including potential biases and methodological constraints, underscore the need for further research to validate these findings and delve deeper into the microbiota's role in HPV development. Despite these limitations, the review provides valuable insights into microbiota trends during cervical cancer progression, offering direction for future research. The review summarizes key findings from previous studies on microbiota during healthy and cancerous stages, as well as other conditions such as CIN, SIL, HPV (+), and HPV (-), indicating a promising area for further investigation. The consistent presence of HPV across all reported cervical abnormalities, along with the identification of distinct bacterial genera between cancerous and control samples, suggests a potential link that merits further exploration. In conclusion, a more profound understanding of the microbial landscape could elucidate the pathogenesis of cervical diseases and inform future strategies for diagnosis, prevention, and treatment.
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AIMS: The aim was to complement the evaluation of functional and safety properties of beneficial vaginal lactic acid bacteria to select the most adequate strains as potential probiotics for their inclusion in a vaginal probiotic formula. METHODS AND RESULTS: Twenty-four beneficial vaginal lactic acid bacteria strains previously isolated from healthy women were characterized by applying phenotypical and genetic techniques. The biofilm formation and exopolysaccharide production by the different strains showed a strong influence of the growth medium on the expression of these properties. The evaluated strains evidenced different antibiotic susceptibility patterns by phenotypic tests, while the resistance genes were not always correlated with the phenotypic profiles. None of the evaluated strains showed lecithinase, gelatinase, or ß-hemolytic activity. Based on the results obtained, Lactobacillus gasseri CRL 1320 and Lacticaseibacillus rhamnosus CRL 1332 were selected to advance in the design of the products. CONCLUSIONS: A wide variability in the properties expressed by beneficial vaginal lactic acid bacteria strains were shown, evidencing a strain-specific and independent behaviour of the species/metabolic group where classified. The selection criteria applied provided a valuable tool for the selection of the best vaginal lactobacilli strains for their inclusion in formulas for vaginal application. SIGNIFICANCE AND IMPACT OF THE STUDY: Since probiotics are an important tool for improving human health, the designed formula are an excellent alternative to reconstitute the vaginal microbiota and prevent/treat urogenital tract infections in women. The results obtained contribute to deepen the characterization of homologous beneficial vaginal lactic acid bacteria and advance in the design of vaginal probiotic products.
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Lactobacillales , Probióticos , Feminino , Humanos , Lactobacillales/genética , Vagina/microbiologia , Antibacterianos , Higiene , Fosfolipases , GelatinasesRESUMO
OBJECTIVE: To compare the diagnosis of bacterial vaginosis (BV) by Nugent scoring criteria (Nugent-BV) and the diagnosis of BV and/or aerobic vaginitis (AV) using Donders criteria (Donders-BV/AV) for identifying Molecular-BV detected by bacterial 16s rRNA profiling. METHODS: We enrolled 512 women of reproductive age in Brazil with data available on Nugent and Donders microscopic analysis and 16S rRNA sequencing. We constructed receiver operating characteristic (ROC) curves of Nugent-BV and Donders-BV/AV and calculated their area under the curves (AUCs) and 95% confidence interval (CI) for matching Molecular-BV. RESULTS: A total of 155 (28.7%) participants were positive for Nugent-BV. Donders-BV and -AV were detected in 90 (17.6%) and 75 (14.6%) participants, respectively, while 28 (5.5%) had concurrent Donders-BV and -AV. Molecular-BV was identified in 139 (27.1%) participants. Analysis of ROC curves showed that diagnosis of Nugent-BV more accurately aligned with presence of Molecular-BV (AUC: 0.88, 95% CI: 0.84-0.91) when compared to Donders-AV/BV (AUC: 0.84; CI: 0.80-0.87) (P = 0.005). CONCLUSION: The use of Nugent-BV is more representative of Molecular-BV than Donders-AV/BV.
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Microbiota , Vaginite , Vaginose Bacteriana , Feminino , Humanos , Microbiota/genética , RNA Ribossômico 16S/genética , Vagina/microbiologia , Vaginite/diagnóstico , Vaginose Bacteriana/diagnóstico , Vaginose Bacteriana/microbiologiaRESUMO
The vaginal microbiota plays vital protection in women. This probiotic activity is caused not only by individual Lactobacillus species but also by its multi-microbial interaction. However, the probiotic activity promoted by multi-microbial consortia is still unknown. The aim of this study was the individual and collective analysis on the prevalence of five vaginal lactobacilli (Lactobacillus iners, Lactobacillus crispatus, Lactobacillus gasseri, Lactobacillus jensenii, and Lactobacillus acidophilus) among healthy women and women with bacterial vaginosis (BV) or aerobic vaginitis (AV). PCR assays were realized on 436 vaginal samples from a previous study. Chi-square, univariable, and multivariable logistic regression analyses with the Benjamini-Hochberg adjustment evaluated associations between these lactobacilli and vaginal microbiota. Multi-microbial clustering model was also realized through Ward's Minimum Variance Clustering Method with Euclidean squared distance for hierarchical clustering to determine the probiotic relationship between lactobacilli and vaginal dysbiosis. Concerning the individual effect, L. acidophilus, L. jensenii, and L. crispatus showed the highest normalized importance values against vaginal dysbiosis (100%, 79.3%, and 74.8%, respectively). However, only L. acidophilus and L. jensenii exhibited statistical values (p = 0.035 and p = 0.050, respectively). L. acidophilus showed a significant prevalence on healthy microbiota against both dysbioses (BV, p = 0.041; and AV, p = 0.045). L. jensenii only demonstrated significant protection against AV (p = 0.012). Finally, our results evidenced a strong multi-microbial consortium by L. iners, L. jensenii, L. gasseri, and L. acidophilus against AV (p = 0.020) and BV (p = 0.009), lacking protection in the absence of L. gasseri and L. acidophilus.
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Vaginose Bacteriana , Vulvovaginite , Análise por Conglomerados , Disbiose , Equador , Feminino , Humanos , Lactobacillus , Lactobacillus acidophilus , Consórcios Microbianos , Vaginose Bacteriana/epidemiologia , Vaginose Bacteriana/microbiologia , Vaginose Bacteriana/prevenção & controleRESUMO
Cervical ectopy is a benign condition of the lower genital tract that is frequently detected in women of reproductive age. Although cervical ectopy is regarded as a physiological condition, some women experience symptoms such as leucorrhoea, persistent bleeding and recurrent vaginal infections that require medical intervention. Cervical ectopy has not been linked to cervical cancer, but it is thought to facilitate the acquisition of sexually transmitted diseases (STDs), like Human Papillomavirus (HPV) infection, as it provides a favorable microenvironment for virus infection and dissemination. We and others have described the presence of oncogenic HPV types in women with symptomatic cervical ectopy. The relevance of this finding and the impact of symptomatic cervical ectopy on the cervicovaginal microenvironment (vaginal microbiota, immune and inflammatory responses) are currently unknown. To shed some light into the interplay between HPV, the vaginal microbiota and mucosal immune and inflammatory responses in the context of this condition, we enrolled 156 women with symptomatic cervical ectopy and determined the presence of HPV using a type-specific multiplex genotyping assay. Overall, HPV was detected in 54.48% women, oncogenic HPV types were found in more than 90% of HPV-positive cases. The most prevalent HPV types were HPV16 (29.4%), HPV31 (21.17%) and HPV18 (15.29%). Next, we evaluated the vaginal microbial composition and diversity by 16S rDNA sequencing, and quantified levels of cytokines and chemokines by flow cytometry using bead-based multiplex assays in a sub-cohort of 63 women. IL-21 and CXCL9 were significantly upregulated in HPV-positive women (p=0.0002 and p=0.013, respectively). Women with symptomatic cervical ectopy and HPV infection had increased diversity (p<0.001), and their vaginal microbiota was enriched in bacterial vaginosis-associated anaerobes (Sneathia, Shuttleworthia, Prevotella, and Atopobium) and depleted in Lactobacillus spp. Furthermore, the vaginal microbiota of women with symptomatic cervical ectopy and HPV infection correlated with vaginal inflammation (IL-1ß, rho=0.56, p=0.0004) and increased mucosal homeostatic response (IL-22, rho=0.60, p=0.0001). Taken together, our results suggest that HPV infection and dysbiotic vaginal communities could favor a vaginal microenvironment that might delay the recovery of the cervical epithelium in women with symptomatic cervical ectopy and favor STDs acquisition.
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Alphapapillomavirus , Microbiota , Infecções por Papillomavirus , Feminino , Humanos , Imunidade nas Mucosas , Masculino , Microbiota/genética , Papillomaviridae/genéticaRESUMO
Introduction. Two high-oncogenic-risk human papilomavirus (hrHPV) genotypes - HPV16 and HPV18 - cause most of the cases of cervical cancer worldwide. Bacterial vaginosis is associated with increased hrHPV persistence, although the mechanism underlying this association remains unclear. Gardnerella spp. are detected in nearly all cases of bacterial vaginosis and are the major source of cervicovaginal sialidases. The NanH1 gene is present in virtually all Gardnerella sialidase-producing strains and has been proposed as a potential marker for persistent hrHPV infection.Hypothesis. Gardnerella spp. load and the NanH1 gene are associated with hrHPV persistence.Aim. To compare the cervicovaginal load of Gardnerella spp. and the frequency of the NanH1 gene between women with persistent HPV16 and/or HPV18 infection and those who cleared the infection after 11 months.Methodology. Among a population of 1638 HPV screened, we detected 104 with positive HPV16 and/or HPV18 results. Samples were obtained at two time points (baseline and at a median of 11 months at follow-up) and tested using the Linear Array HPV Genotyping kit (Roche Molecular Systems, Pleasanton, CA, USA). Based on their HPV16/HPV18 status at enrolment and follow-up, participants were assigned to 'persistence' or 'clearance' groups. We used cervicovaginal fluid samples obtained upon enrolment to determine the load of the 23 s rRNA gene of Gardnerella spp. and the presence of the NanH1 gene using real-time polymerase chain reaction (PCR). We compared Gardnerella spp. loads and NanH1 frequency between the groups by, respectively, Mann-Whitney and chi-squared tests, with a P-value <0.05 considered to be significant.Results. Of the 104 participants who were positive for HPV16/HPV18, 73 (70.2â%) persisted with at least 1 of the baseline genotypes at follow-up, while 31 (29.8â%) cleared the infection in this time frame. Participants in the persistence group had significantly higher loads of Gardnerella spp. [5.8E+02 (0-3.0E+05) copies µl-1] than those in the clearance group [9.9E+01 (0-7.7E+04) copies µl-1] (P=0.03). The baseline frequency of NanH1 was higher in the persistence' (n=46, 63.0â%) than in the clearance (n=14, 45.2â%) group, although this was not statistically significant (P=0.09).Conclusion. These findings reinforce the negative effect of vaginal microbiota for the clearance of hrHPV and indicate a possible association between sialidase-producing species with hrHPV persistence.
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Infecções por Papillomavirus , Neoplasias do Colo do Útero , Vaginose Bacteriana , Feminino , Gardnerella/genética , Papillomavirus Humano 16/genética , Papillomavirus Humano 18/genética , Humanos , NeuraminidaseRESUMO
BACKGROUND: The vaginal microenvironment, regulated by an immune system, can be protected or altered by many factors, including contraceptive methods. OBJECTIVE: The objective of this study is to evaluate the impact of contraceptive methods on the basic vaginal states (BVSs) and to identify culturable vaginal Lactobacillus species. METHODS: This is a prospective, consecutive, longitudinal, and descriptive study. The vaginal contents of 208 women were sampled prior to initiating contraception and six months later. The BVSs were established using the balance of vaginal content (BAVACO) methodology that evaluates microbiota and vaginal inflammatory reaction (VIR). Lactobacillus species were characterized by biochemical tests and mass spectrometry. The following contraceptive methods were evaluated: combined oral contraceptive pill (COCP), condom (CON) and rhythm method (RHYT). McNemar's test was used. RESULTS: Of the 208 women, 171 attended both examinations. In the COCP group (n=127), 83 vaginal contents maintained a normal microbiota, 1 sample became dysbiotic, and 37/43 dysbiotic microbiota samples reverted to normal (p<0.0001). A conversion to BVS with VIR was detected in the CON group (n=31) (p=0.001). The RHYT group (n=13) maintained its initial BVSs. The predominant Lactobacillus species found were L. crispatus and L. gasseri, with a trend toward a positive association between L. crispatus and COCP (OR=2.82; p=0.058). CONCLUSION: Hormone administration corrected the dysbiosis and preserved a normal BVS. The CON increased the VIR. The protection of the microbiota observed in the rhythm method probably responds to a systemic hormonal influence. The trend toward a positive association between COCP and L. crispatus, with its protective properties, evidenced an effective hormonal relationship.
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Anticoncepcionais Orais Combinados , Lactobacillus , Anticoncepção , Anticoncepcionais Orais Combinados/metabolismo , Feminino , Humanos , Lactobacillus/metabolismo , Estudos Prospectivos , VaginaRESUMO
Resumen Introducción: Las infecciones de transmisión sexual (ITS) han incrementado su incidencia universalmente en la última década, incluido Chile. Una de las poblaciones afectadas es la privada de libertad. Objetivo: Evaluar la prevalencia ITS en mujeres del Centro de Detención Preventiva de Arica y Parinacota (Chile) y su asociación con factores biodemográficos. Metodología: En 127 mujeres se realizó un análisis bivariado de los resultados de serología para VHB, VHC, VIH1-2 y VDRL, y un estudio de flujo vaginal convencional microbiológico incluyendo Trichomonas vaginalis, Neisseria gonorrhoeae y Chlamydia trachomatis. Resultados: El 33,1% de las reclusas tuvo al menos una ITS; aquellas menores de 34 años, las consumidoras de drogas y con más de una pareja sexual tuvieron mayor riesgo. Las ITS prevalentes fueron infección por VIH (15,7%) y sífilis (7,9%) asociadas al consumo de drogas y relaciones sexuales antes de 14 años. Trichomonas vaginalis (12,9%) se encontró en mujeres jóvenes con más de una pareja sexual. El 53,2% tuvo un cultivo de flujo vaginal positivo, principalmente con Gardenella vaginalis (32,3%), asociada al mayor número de parejas sexuales y menor tiempo de estadía en reclusión. Candida albicans (11,3%) tuvo mayor prevalencia en mujeres entre 19 y 24 años no heterosexuales. Chlamydia trachomatis, VHB, VHC y N. gonorrhoeae tuvieron prevalencias menores. Conclusión: Existe una alta frecuencia de infección por VIH, sífilis y T vaginalis, predominio de G. vaginalis en aproximadamente un tercio de las mujeres estudiadas y en sobre la mitad de los casos estudiados se comprobó una disbiosis vaginal.
Abstract Background: Sexually transmitted infections (STIs) have increased their incidence worldwide in the last decade, as well as in Chile. One of the affected populations is the deprived of liberty. Aim: To evaluate the STI prevalence in women from the Arica y Parinacota Preventive Detention Center (Chile) and its association with biodemographic factors. Methods: 127 women were studied who underwent a bivariate analysis of the serology results for HBV, HCV, HIV1-2 and VDRL, and a study of conventional microbiological vaginal discharge including Trichomonas vaginalis, Neisseria gonorrhoeae and Chlamydia trachomatis. Results: 33.1% of the inmates had at least one STI, where, women under 34 years old, drug use and more than one sexual partner were at greater risk. The most prevalent STI were HIV infection (15.7%) and syphilis (7.9%) associated with drug use and sexual intercourse before the age of14. Trichomonas vaginalis (12.9%) was identified in young women with more than one sexual partner. 53.2% had a positive culture, mainly with Gardenella vaginalis (32.3%) associated with an increase in sexual partners and a shorter stay in prison. Candida albicans (11.3%) had a higher prevalence in non-heterosexual women between 19 and 24 years old. Chlamydia trachomatis, HBV, HCV and N. gonorrhoeae had lower prevalences. Conclusion: There is a high frequency of HIV infection, syphilis and T. vaginalis, predominance of G. vaginalis in approximately a third of the women studied and about half of the cases studied had vaginal dysbiosis.
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Cervical cancer is an important health concern worldwide and is one of the leading causes of death in Mexican women. Previous studies have shown changes in the female genital tract microbe community related to human papillomavirus (HPV) infection and cervical cancer; yet, this link remains unexplored in many human populations. This study evaluated the vaginal bacterial community among Mexican women with precancerous squamous intraepithelial lesions (SIL). We sequenced the V3 region of the 16S rRNA gene in cervical samples from 228 Mexican women, including 121 participants with SIL, most of which were HPV positive, and 107 healthy women without HPV infection or SIL. The presence of SIL was associated with changes in composition (beta diversity) and with a higher species richness (Chao1). A comparison of HPV-positive women with and without SIL showed that microbiota changes occurred even in the absence of SIL. Multivariate association with linear models (MaAsLin) analysis yielded independent associations between HPV infection and an increase in the relative abundance of Brachybacterium conglomeratum and Brevibacterium aureum as well as a decrease in two Lactobacillus iners operational taxonomic units (OTUs). We also identified a positive independent association between HPV-16, the most common HPV subtype linked to SIL, and Brachybacterium conglomeratum. Our work indicates that HPV infection leading to SIL is primarily associated with shifts in vaginal microbiota composition, some of which may be specific to this human population. IMPORTANCE Human papillomavirus (HPV) plays a critical role in cervical carcinogenesis but is not sufficient for cervical cancer development, indicating the involvement of other factors. The vaginal microbiota is an important factor in controlling infections caused by HPV, and, depending on its composition, it can modulate the microenvironment in vaginal mucosa against viral infections. Ethnic and sociodemographic factors influence differences in vaginal microbiome composition, which underlies the dysbiotic patterns linked to HPV infection and cervical cancer across different populations of women. Here, we provide evidence for associations between vaginal microbiota patterns and HPV infection linked to ethnic and sociodemographic factors. To our knowledge, this is the first report of the species Brevibacterium aureum and Brachybacterium conglomeratum linked to HPV infection or squamous intraepithelial lesions (SIL).
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Bactérias/classificação , Microbiota/genética , Displasia do Colo do Útero/patologia , Neoplasias do Colo do Útero/patologia , Vagina/microbiologia , Actinobacteria/genética , Actinobacteria/isolamento & purificação , Adulto , Alphapapillomavirus , Bactérias/genética , Bactérias/isolamento & purificação , Brevibacterium/genética , Brevibacterium/isolamento & purificação , Disbiose/microbiologia , Células Epiteliais/patologia , Feminino , Humanos , Lactobacillus/genética , Lactobacillus/isolamento & purificação , México , Infecções por Papillomavirus/patologia , RNA Ribossômico 16S/genética , Determinantes Sociais da Saúde , Fatores Sociodemográficos , Neoplasias do Colo do Útero/microbiologia , Neoplasias do Colo do Útero/virologia , Displasia do Colo do Útero/microbiologia , Displasia do Colo do Útero/virologiaRESUMO
The profitability of commercial pig farms largely depends on the reproductive performance of gilts and sows. The aim of this study was to identify differences in the composition and diversity of vaginal microbiota between gilts (G) and pregnant (P) sows, both artificially inseminated (AI) and natural mating (NM). Samples were collected by scraping the vaginal mucosa of G (n = 10) and P (NM, n = 10 and AI, n = 7) sows. Samples were analysed by culture-dependent techniques and 16S-rRNA gene High-Throughput-Sequencing. The profiles of the cultured microbiota showed two distinctive clusters, one of them grouped four samples of P sows from the AI group. The vaginal microbiota from P had lower richness than G sows (Mann-Whitney/Kruskal-Wallis test, p < 0.01), but all vaginal samples had a similar diversity. The PERMANOVA analyses revealed significant differences (p < 0.01) between the microbial communities' structures from G and P sows. The bacteria phyla with the highest relative abundances were Proteobacteria (33.1%), followed by Firmicutes (32%), Cyanobacteria (13.3%) and Actinobacteria (13.2%). The relative abundance for phyla, families and genera was estimated and Proteobacteria was significantly higher (p = 0.038) in P than in G sows; Firmicutes was significantly lower in AI than G and NM sows. A "core microbiota" included Lactobacillus, Bacillus, Enterococcus, Acinetobacter and Pseudomonas. The results presented highlight the differences in the bacterial composition between G and P sows, as well as the changes in the microbial populations associated with the breeding method.
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Inseminação Artificial , Reprodução , Animais , Bactérias/genética , Feminino , Inseminação Artificial/veterinária , Gravidez , RNA Ribossômico 16S/genética , Sus scrofa , Suínos , VaginaRESUMO
INTRODUCTION: Sialidase activity in the cervicovaginal fluid (CVF) is associated with microscopic findings of bacterial vaginosis (BV). Sequencing of bacterial 16S rRNA gene in vaginal samples has revealed that the majority of microscopic BV cases fit into vaginal community-state type IV (CST IV), which was recently named "molecular-BV." Bacterial vaginosis-associated bacterial species, such as Gardnerella spp., may act as sources of CVF sialidases. These hydrolases lead to impairment of local immunity and enable bacterial adhesion to epithelial and biofilm formation. However, the impact of CVL sialidase on microbiota components and diversity remains unknown. OBJECTIVE: To assess if CVF sialidase activity is associated with changes in bacterial components of CST IV. METHODS: One hundred forty women were cross-sectionally enrolled. The presence of molecular-BV (CST IV) was assessed by V3-V4 16S rRNA sequencing (Illumina). Fluorometric assays were performed using 2-(4-methylumbelliferyl)-α-D-N-acetylneuraminic acid (MUAN) for measuring sialidase activity in CVF samples. Linear discriminant analysis effect size (LEfSe) was performed to identify the differently enriched bacterial taxa in molecular-BV according to the status of CVF sialidase activity. RESULTS: Forty-four participants (31.4%) had molecular-BV, of which 30 (68.2%) had sialidase activity at detectable levels. A total of 24 bacterial taxa were enriched in the presence of sialidase activity, while just two taxa were enriched in sialidase-negative samples. CONCLUSION: Sialidase activity in molecular-BV is associated with changes in bacterial components of the local microbiome. This association should be further investigated, since it may result in diminished local defenses against pathogens.
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Microbiota , Vaginose Bacteriana , Feminino , Humanos , Lactobacillus/genética , Neuraminidase/genética , RNA Ribossômico 16S/genética , Vagina/microbiologia , Vaginose Bacteriana/microbiologiaRESUMO
Pregnancy outcomes and women's health are directly affected by vaginal microbiota. This microbiota consists of a dynamic ecosystem of various microbes in different ratios, which in healthy conditions protect the vaginal epithelium from infections. However, cases of vaginal infection are regularly diagnosed in women of reproductive age, contributing to more severe outcomes. Therefore, our main goal was to determine the prevalence of bacterial vaginosis (BV), aerobic vaginitis (AV), and vulvovaginal candidiasis (VVC) among Ecuadorian pregnant and non-pregnant women. A cross-sectional study was conducted among 217 women between 13 and 40 years old seeking primary healthcare in Carlos Andrade Marin Hospital (HCAM), Gynecological-Obstetric Hospital Isidro Ayora (HGOIA) and Center for Teaching Health Cipriana Dueñas during October 2018 to February 2019. The classical characterization of the vaginal microbiota was performed through microscopy by the Nugent criteria to evaluate the presence of BV, healthy and intermediate microbiota, by the criteria of Donders to determine the presence of AV and by the Marot-Leblond criteria to diagnose VVC. DNA extraction from vaginal samples and Polymerase Chain Reaction (PCR) analysis was performed to characterize the presence of Gardnerella spp., Mobiluncus mulieris, Escherichia coli, Enterococcus spp., and Lactobacillus spp. Finally, quantification of the lactobacilli was performed by quantitative real-time PCR (qPCR) for samples from women with normal vaginal microbiota and women with AV. Our results showed 52% of women with healthy microbiota, 7% with intermediate microbiota, and 41% with vaginal dysbiosis, comprising 27% with AV, 8% with BV and 4% with VVC and 2% with co-infections or co-dysbiosis. Additionally, a higher amount of lactobacilli were found in pregnant women when compared to non-pregnant women, while AV cases were characterized by a significant drop of Lactobacillus spp., more precisely, between 1E3 and 1E5 colony forming units (CFU)/ml. Finally, women with normal vaginal microbiota showed an average load of lactobacilli between 1E6 and 1E7 CFU/ml. This pilot study showed no statistically significant differences between pregnant and non-pregnant women, pointing to the possibility to use lactobacilli quantification for the prevention of future vaginal infections.
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Lactobacillus , Microbiota , Adolescente , Adulto , Estudos Transversais , Feminino , Humanos , Lactobacillus/genética , Projetos Piloto , Gravidez , Reação em Cadeia da Polimerase em Tempo Real , Vagina , Adulto JovemRESUMO
The characterization of vaginal microbiota will help to understand some of the reproductive problems and mechanisms to improve cattle reproduction. The objective of this study was to characterize the vaginal microbiota of cyclical Holstein cows with different parturition orders using 16S rDNA sequencing. Animals were submitted to an estrus synchronization protocol with the use of intravaginal progesterone (P4) implants and were treated or not with ceftiofur hydrochloride. DNA samples were extracted from vaginal swabs on day 0 and 10 of the synchronization, and sequenced with the Illumina MiSeq platform with an average coverage rate of 10.000 reads per samples using a Single-End library for fragments of 300 bp. The main bacterial phyla found in the vaginal tract of Holstein cows, were Firmicutes (37.61%), Tenericutes (29.45%), Proteobacteria (17.47%) and Bacteriodetes (13.73%), followed by Actinobacteria (0.82%) and Spirochaetae (0.45%). The use of intravaginal P4 devices has increased the relative abundance of the genera Family XIII AD3011 and Family XIII unclassified (p < .049). We have also observed an effect of the number of calving on the vaginal microbiota composition, showing that multiparous cows have a greater bacterial diversity than primiparous animals (p < .05). The use of ceftiofur hydrochloride was effective to reduce the vaginal bacteria proliferation. This study describes for the first time the vaginal microbiota of cows synchronized with intravaginal progesterone devices, different from the traditional methods such as microbiological culture and biochemical tests. We have identified a large number of microorganisms commonly found in the gastrointestinal tract of cows, colonizing the vaginal microbiota.
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Bovinos , Sincronização do Estro/métodos , Microbiota/efeitos dos fármacos , Progesterona/farmacologia , Vagina/microbiologia , Administração Intravaginal , Animais , Feminino , Inseminação Artificial/veterinária , Progesterona/administração & dosagem , Progestinas/administração & dosagem , Progestinas/farmacologia , ReproduçãoRESUMO
Vaginal drug delivery represents an attractive alternative to achieve local and systemic effects due to the high contact surface exposed, the mucoadhesion of the epithelium, and the high innervation that facilitates the absorption of drugs into the bloodstream. However, despite the confinement of the vaginal cavity, it is an organ with a highly variable microenvironment. Mechanical alterations such as coitus, or chemical changes such as pH and viscosity, modify the release of drugs. In addition, changes in vaginal microbiota can influence the entire vaginal microenvironment, thus determining the disposition of drugs in the vaginal cavity and decreasing their therapeutic efficacy. Therefore, the influence of microorganisms on vaginal homeostasis can change the pre-established scenario for the application of drugs. This review aims to provide an explanation of normal vaginal microbiota, the factors that modify it, its involvement in the administration of drugs, and new proposals for the design of novel pharmaceutical dosage forms. Finally, challenges and opportunities directed toward the conception of new effective formulations are discussed.
RESUMO
In the context of HIV sexual transmission at the genital mucosa, initial interactions between the virus and the mucosal immunity determine the outcome of the exposure. Hence, these interactions have been deeply explored in attempts to undercover potential targets for developing preventative strategies. The knowledge gained has led to propose a hypothetical model for mucosal HIV transmission. Subsequent research studies on this topic further revealed new mechanisms and identified new host-HIV interactions. This review aims at integrating these findings to inform better and update the current model of HIV transmission. At the earliest stage of virus exposure, the epithelial integrity and the presence of antiviral factors are critical in preventing viral entry to the submucosa. However, the virus has been shown to enter to the submucosa in the presence of physical abrasion or via epithelial transmigration using paracellular passage or transcytosis mechanisms. The efficiency of these processes is greater with cell-associated viral inoculums and can be influenced by the presence of viral and immune factors, and by the structure of the exposed epithelium. Once the virus reaches the submucosa, dendritic cells and fibroblasts, as recently described, have been shown in vitro of being capable of facilitating the transfer of viral particles to susceptible cells, leading to viral dissemination, most likely in a trans-infection manner. The presence of activated CD4+ T cells in submucosa increases the probability of infection, where the predominant microbiota could be implicated through the modulation of an inflammatory microenvironment. Other factors such as genital fluids and hormones could also play an essential role in HIV transmission. Here, we review the most recent evidence described for mucosal HIV-transmission contributing with the understanding of this phenomenon.
Assuntos
Infecções por HIV/transmissão , Mucosa/virologia , Animais , Genitália/imunologia , Infecções por HIV/imunologia , Humanos , Imunidade nas Mucosas , Mucosa/imunologiaRESUMO
BACKGROUND: The vaginal microbiome is influenced by a wide variety of factors, including contraceptive methods. AIM: To evaluate the effect of contraceptive methods on vaginal microbiota and to compare MALDI-TOF MS and 16S rDNA sequencing for lactobacilli identification. PATIENTS AND METHODS: One hundred and one (101) women consulting for birth control were included in a prospective study. Their vaginal content was sampled and analyzed once before they started using the contraceptive method of their choice, and twice after the initiation of contraception, at three months (94/101 women attended) and at six months (89/101 women attended). The relative frequencies of yeasts and trichomonas were analyzed. MALDI-TOF MS and 16S rDNA sequence analysis were applied for the identification of lactobacilli in their vaginal microbiota. The following contraceptive methods were assessed: Combined Oral Contraceptive Pill (COCP), Condom (CON) and The Rhythm Method (RHYT). McNemar's statistical test was applied. RESULTS: A statistically significant association between COCP and normal microbiota was observed after three months (p< 0.01) and after six months (p< 0.0001), when the vaginal microbiota was modified. At six months, inflammatory reaction was detected in 3/7 women in the CON group, while 6/7 patients using RHYT showed the same state. Yeast colonization increased with the COCP. Identification of lactobacilli by MALDI-TOF MS analysis compared to 16S rDNA sequencing yielded 92.9% concordant results. Lactobacillus gasseri and L. crispatus were the predominant species. CONCLUSION: The pattern of vaginal states was significantly modified. Hormone administration apparently corrected the alterations and retained a normal vaginal state. MALDI-TOF MS has the potential of being an accurate tool for the identification of vaginal lactobacilli species L. murinus was for the first time isolated from the vagina.