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1.
BJOG ; 127(2): 159-169, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31376240

RESUMO

OBJECTIVES: To resolve the controversy regarding the presence of a microbiota in the placenta. DESIGN: Classical and molecular microbiological study. SETTING: All samples were collected during caesarean section. POPULATION: A total of 28 human placentas and six murine placentas. METHODS: All 28 human placentas were checked for 16S rRNA gene amplification products. Three locations from four selected human placentas and three 'environmental controls' for each placenta were placed in seven culture media. The four selected human placentas were further analysed using Gram stain, immunohistochemistry for bacteria, electron microscopy, and TaqMan RT-qPCR. Six placentas from three SPF mice were cut into four pieces each, and further analysed for 16S rRNA gene amplification. MAIN OUTCOME MEASURES: Microbiological and molecular evidence of bacteria. RESULTS: None of the placental cultures used for the full analysis, or their environmental cultures, was positive for bacterial growth. None of the other methods showed any evidence of bacteria. Immunohistochemistry showed negligible bacterial counts. None of the murine placentas showed evidence of 16S rRNA gene amplification. CONCLUSIONS: Our results support that the fetal environment in the womb is sterile. Based on the immunohistochemistry and the limit of detection of the other methods used, if a placental microbiome exists, it is of extreme low biomass, and thus its effect on clinical phenotypes is probably minor, if it exists at all. TWEETABLE ABSTRACT: Using several microbiological and molecular methods in parallel, we found no compelling evidence of bacteria in human and mouse placentas.


Assuntos
Líquido Amniótico/microbiologia , Microbioma Gastrointestinal/fisiologia , Microbiota/genética , Placenta/microbiologia , RNA Ribossômico 16S/fisiologia , Líquido Amniótico/imunologia , Animais , Feminino , Microbioma Gastrointestinal/imunologia , Humanos , Imuno-Histoquímica , Metagenômica , Camundongos , Placenta/imunologia , Gravidez , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
2.
BJOG ; 127(2): 147-158, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31587490

RESUMO

The female reproductive tract represents a continuum between the vagina and the upper genital tract. New evidence from cultivation-independent studies suggests that the female upper genital tract is not sterile; however, the significance of this for reproductive health and disease remains to be elucidated fully. Further, diagnosis and treatment of infectious reproductive tract pathologies using cultivation-independent technologies represents a largely unchartered area of modern medical science. The challenge now is to design well-controlled experiments to account for the ease of contamination known to confound molecular-based studies of low-biomass niches, including the uterus and placenta. This will support robust assessment of the potential function of microorganisms, microbial metabolites, and cell-free bacterial DNA on reproductive function in health and disease. TWEETABLE ABSTRACT: Molecular microbial studies of low-biomass niches require stringent experimental controls to reveal causal relations in reproductive health and disease.


Assuntos
Biomassa , Microbioma Gastrointestinal/fisiologia , Placenta/microbiologia , Saúde Reprodutiva , Infecções do Sistema Genital/microbiologia , Vagina/microbiologia , Feminino , Microbioma Gastrointestinal/genética , Microbioma Gastrointestinal/imunologia , Humanos , Metagenômica , Placenta/imunologia , Gravidez , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Vagina/imunologia
3.
BMC Infect Dis ; 19(1): 1001, 2019 Nov 27.
Artigo em Inglês | MEDLINE | ID: mdl-31775645

RESUMO

BACKGROUND: Q fever has been associated with perinatal complications. We conducted a prospective follow-up study to assess both the incidence of adverse pregnancy outcomes (APOs) associated with Coxiella burnetii infection and the contribution of Q fever to APOs. METHODS: Between May 1 and October 31, 2013, within the regional perinatal health care centre of Saint Pierre, Reunion island, we investigated unexplained miscarriages, stillbirths, preterm births or small-for-gestational age children. Seropositivity for C. burnetii antibodies was defined using indirect immunofluorescence for a phase 2 IgG titre ≥1:64. Acute Q fever was defined for a high phase 2 IgG titre ≥1:256 (compatible with recent or active infection) or the detection of C. burnetii genome in miscarriage products and placentas. Incidence rate ratios (IRR) for Q fever related APOs (taken as a composite outcome or individually) were assessed using Poisson regression models for dichotomous outcomes controlling major confounders. RESULTS: Over a 6-month period, 179 pregnant women suspected or diagnosed with an APO were investigated for Q fever, of whom 118 met the definition for an APO. Of these, 19 were seropositive and 10 presented a profile indicative of an acute infection. For three women with an acute Q fever, the chronology between the onset of infection, the APO (2 miscarriages, 1 preterm birth) and the seroconversion suggested causality in the pathogenesis. The cumulative incidence of Q fever related APOs was estimated between 2.2‰ and 5.2‰, whether causality was required or not. Both C. burnetii exposure and acute Q fever were independently associated with APOs (IRR 1.55, 95% CI 1.31-1.84; IRR 1.47, 95% CI 1.15-1.89, respectively). CONCLUSIONS: In the endemic context of Reunion island, acute Q fever may lead to APOs. To limit the burden of Q fever on reproduction, pregnant women should be kept away from farms and avoid direct contact with ruminants.


Assuntos
Coxiella burnetii/genética , Coxiella burnetii/imunologia , Complicações Infecciosas na Gravidez/epidemiologia , Resultado da Gravidez/epidemiologia , Febre Q/epidemiologia , Adolescente , Adulto , Anticorpos Antibacterianos/imunologia , Coxiella burnetii/isolamento & purificação , DNA Bacteriano/genética , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Seguimentos , Humanos , Incidência , Placenta/microbiologia , Gravidez , Estudos Prospectivos , Reação em Cadeia da Polimerase em Tempo Real , Estudos Retrospectivos , Reunião/epidemiologia , Adulto Jovem
4.
Int J Mol Sci ; 20(14)2019 Jul 11.
Artigo em Inglês | MEDLINE | ID: mdl-31373310

RESUMO

Bacteria colonize most of the human body, and the female genital tract is not an exception. While the existence of a vaginal microbiota has been well established, the upper genital tract has been considered a sterile environment, with a general assumption that bacterial presence is associated with adverse clinical manifestation. However, recent metagenomic studies identified specific patterns of microbiota colonizing the uterus, fallopian tubes, ovaries, and placenta. These results need confirmation and further investigations since the data are only scarce. Bacterial colonization of these sites appears different from the vaginal one, despite evidence that vaginal bacteria could ascend to the upper genital tract through the cervix. Are these bacteria only commensal or do they play a role in the physiology of the female upper genital tract? Which are the genera that may have a negative and a positive impact on the female reproductive function? The aim of this review is to critically present all available data on upper genital tract microbiota and discuss its role in human reproduction, ranging from the technical aspects of these types of analyses to the description of specific bacterial genera. Although still very limited, research focusing on genital colonization of bacteria other than the vaginal milieu might bring novel insights into physiopathology of human reproduction.


Assuntos
Tubas Uterinas/microbiologia , Lactobacillus/isolamento & purificação , Ovário/microbiologia , Placenta/microbiologia , Útero/microbiologia , Bacteroidetes/genética , Bacteroidetes/isolamento & purificação , Feminino , Humanos , Lactobacillus/genética , Microbiota/fisiologia , Gravidez , Proteobactérias/genética , Proteobactérias/isolamento & purificação , RNA Ribossômico 16S/genética
5.
Nature ; 572(7769): 329-334, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31367035

RESUMO

We sought to determine whether pre-eclampsia, spontaneous preterm birth or the delivery of infants who are small for gestational age were associated with the presence of bacterial DNA in the human placenta. Here we show that there was no evidence for the presence of bacteria in the large majority of placental samples, from both complicated and uncomplicated pregnancies. Almost all signals were related either to the acquisition of bacteria during labour and delivery, or to contamination of laboratory reagents with bacterial DNA. The exception was Streptococcus agalactiae (group B Streptococcus), for which non-contaminant signals were detected in approximately 5% of samples collected before the onset of labour. We conclude that bacterial infection of the placenta is not a common cause of adverse pregnancy outcome and that the human placenta does not have a microbiome, but it does represent a potential site of perinatal acquisition of S. agalactiae, a major cause of neonatal sepsis.


Assuntos
Parto Obstétrico , Complicações do Trabalho de Parto/microbiologia , Placenta/microbiologia , Complicações Infecciosas na Gravidez/microbiologia , Sepse/congênito , Sepse/microbiologia , Streptococcus agalactiae/isolamento & purificação , Streptococcus agalactiae/patogenicidade , Biópsia , Estudos de Coortes , Contaminação por DNA , DNA Bacteriano/análise , DNA Bacteriano/genética , DNA Bacteriano/isolamento & purificação , Feminino , Humanos , Recém-Nascido , Masculino , Metagenômica , Gravidez , Resultado da Gravidez , RNA Ribossômico 16S/análise , RNA Ribossômico 16S/genética , Reprodutibilidade dos Testes , Análise de Sequência de DNA
6.
Infect Immun ; 87(10)2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31331955

RESUMO

Porphyromonas gingivalis is considered a keystone pathogen that contributes to the initiation and progression of periodontitis in humans. P. gingivalis has also been detected in human placentas associated with adverse pregnancy outcomes. The spread of P. gingivalis from the oral cavity to the reproductive tract thus represents a potential mechanism whereby periodontitis can lead to adverse pregnancy outcomes. In a murine model of pregnancy and oral infection with P. gingivalis, C57BL/6J mice developed low fetal weight, whereas C57BL/6NCrl mice did not. Although C57BL/6NCrl mice harbor segmented filamentous bacteria that drive a Th17 response, fetal weight was independent of frequency of Th17 or Th1 in either substrain. Low fetal weight was instead correlated with increasing amounts of P. gingivalis DNA in the placentas of the C57BL/6J dams. In contrast, fetal weight in C57BL/6NCrl mice was independent of P. gingivalis in the placenta. Differences in genetics or microbiome that influence the ability of P. gingivalis to colonize the placenta may drive differential fetal weight outcomes between C57BL/6J and C57BL/6NCrl mice and, potentially, between diverse human populations.


Assuntos
Infecções por Bacteroidaceae/microbiologia , Peso Fetal , Periodontite/microbiologia , Porphyromonas gingivalis/patogenicidade , Complicações Infecciosas na Gravidez/microbiologia , Células Th17/microbiologia , Animais , Infecções por Bacteroidaceae/imunologia , Infecções por Bacteroidaceae/patologia , Modelos Animais de Doenças , Feminino , Feto , Expressão Gênica , Interferon gama/genética , Interferon gama/imunologia , Interleucina-17/genética , Interleucina-17/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Boca/imunologia , Boca/microbiologia , Periodontite/imunologia , Periodontite/patologia , Placenta/imunologia , Placenta/microbiologia , Porphyromonas gingivalis/imunologia , Gravidez , Complicações Infecciosas na Gravidez/imunologia , Complicações Infecciosas na Gravidez/patologia , Especificidade da Espécie , Células Th17/imunologia
7.
BMC Vet Res ; 15(1): 257, 2019 Jul 24.
Artigo em Inglês | MEDLINE | ID: mdl-31340816

RESUMO

BACKGROUND: Infectious abortion in ruminants is a problem in animal husbandry worldwide. It is important to obtain a diagnosis, to make sure that proper control measures can be instituted, but most abortion cases remain without an etiologic diagnosis. This report describes the presence of Arcobacter species and several neglected opportunistic abortifacient agents in ruminant abortion cases showing or not co-infections among at least one of the major recognized protozoal, fungal, bacterial and viral abortifacient agents. RESULTS: A total of 67 fetuses (55 cattle and 12 goats) and just one placenta (cattle) were considered. Among the most common abortive agents, Neospora caninum (19,4%), followed by Chlamydophila abortus (4,5%), Listeria monocytogenes 1/2a (2,98%), Bovine Viral Diarrhea Virus type 1b (2,98%), Bovine herpesvirus 4 (2,98%), and Aspergillus spp. (2,98%) were detected. The isolated neglected opportunistic bacteria include Escherichia coli, Acinetobacter lwoffii, Staphylococcus spp., Streptococcus spp., Streptococcus uberis, Streptococcus suis, Trueperella pyogenes, Mannheimia haemolytica, Bacillus cereus and Nocardia spp. Other bacterial species, not associated with abortion by literature, but described as causes of diseases occurring sporadically both in humans and animals, were also detected. Three Arcobacter strains, namely two A. skirrowii and one A. cryaerophilus, were isolated from 3 bovine aborted fetuses, and A. butzleri was isolated from the placenta. CONCLUSIONS: A not negligible isolation of Arcobacter species and other neglected abortifacient agents has to be mentioned, with prevalences that seem to be emerging and replacing or co-placing the major infectious players in bovine and caprine reproductive failure due to abortion disease, even if further studies investigating the aetiological power and transmission routes are needed in order to define the role of these microrganisms in ruminant abortion.


Assuntos
Feto Abortado/microbiologia , Feto Abortado/parasitologia , Feto Abortado/virologia , Arcobacter/isolamento & purificação , Doenças dos Bovinos/epidemiologia , Doenças das Cabras/epidemiologia , Infecções Oportunistas/veterinária , Aborto Animal/epidemiologia , Aborto Animal/microbiologia , Aborto Animal/parasitologia , Aborto Animal/virologia , Animais , Arcobacter/classificação , Infecções Bacterianas/epidemiologia , Infecções Bacterianas/veterinária , Bovinos , Doenças dos Bovinos/microbiologia , Doenças dos Bovinos/parasitologia , Doenças dos Bovinos/virologia , Feminino , Doenças das Cabras/microbiologia , Doenças das Cabras/parasitologia , Doenças das Cabras/virologia , Cabras , Itália/epidemiologia , Micoses/epidemiologia , Micoses/veterinária , Doenças Parasitárias em Animais/epidemiologia , Placenta/microbiologia , Gravidez , Viroses/epidemiologia , Viroses/veterinária
8.
Emerg Infect Dis ; 25(8): 1604-1607, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31310204

RESUMO

Whether Mycobacterium leprae transmits from placenta to fetus remains unknown. We describe the case of a pregnant woman with untreated histoid leproma. Although her newborn was healthy, laboratory examination revealed intact M. leprae present in the placenta, suggesting that the placental barrier might prevent vertical dissemination of M. leprae.


Assuntos
Hanseníase/diagnóstico , Hanseníase/microbiologia , Mycobacterium leprae/isolamento & purificação , Placenta/microbiologia , Complicações Infecciosas na Gravidez/diagnóstico , Complicações Infecciosas na Gravidez/microbiologia , Antituberculosos/uso terapêutico , Biópsia , China/epidemiologia , Feminino , Humanos , Hanseníase/epidemiologia , Gravidez , Complicações Infecciosas na Gravidez/epidemiologia , Resultado da Gravidez , Pele/patologia , Resultado do Tratamento , Ultrassonografia
9.
Zhonghua Yu Fang Yi Xue Za Zhi ; 53(6): 633-637, 2019 Jun 06.
Artigo em Chinês | MEDLINE | ID: mdl-31177764

RESUMO

The role of microbes in regulating the metabolism and immune function of the body has been highlighted. The interactions of host-microbe and microbial-microbe have existed before the birth. However, the existence of placental microbiota and their regulatory effects on pregnancy are still controversial. It was found that placental microbes might be implanted into the placenta via urogenital-placental, gastrointestinal-placental, and oral-placental routes. Microbiome may play a role in promoting the outcome of benign pregnancy, and the placental microbiota in pathological pregnancy may be associated to adverse pregnancy outcome. Placenta and amniotic fluid microbes have been linked to abortion, chorioamnionitis, premature rupture of membranes, premature delivery and stillbirth in Cultivation-dependent and Cultivation-independent studies. Obesity, gestational diabetes mellitus, probiotics and antibiotics during pregnancy may be the main factors affecting placenta microbiology.


Assuntos
Corioamnionite , Microbiota , Placenta , Nascimento Prematuro , Líquido Amniótico , Feminino , Humanos , Placenta/microbiologia , Gravidez
10.
J Vet Diagn Invest ; 31(4): 634-639, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31179891

RESUMO

A cluster of 4 bovine abortions caused by Coxiella burnetii occurred in a dairy herd in Uruguay during a 2-mo period. Case 1 consisted of a placenta from an aborted cow; cases 2-4 were fetuses and their placentas. Grossly, the placenta from one aborted cow had moderate, diffuse reddening of the cotyledons and loss of translucency of the intercotyledonary areas. No gross lesions were observed in the other 3 placentas. Microscopically, 2 of 4 placentas had fibrinonecrotizing placentitis with abundant intratrophoblastic gram-negative coccobacilli. C. burnetii was identified intralesionally by immunohistochemistry (IHC) in all 4 placentas, and by PCR and DNA sequencing in 3 placentas analyzed by these techniques. One fetus had mild neutrophilic alveolitis with multinucleate syncytial cells; no gross or microscopic lesions were observed in the other 2 fetuses examined. The lungs of the 3 fetuses were negative for C. burnetii by IHC. Tests performed to investigate other possible causes of abortions in the 4 cases were negative. C. burnetii causes Q fever in humans and coxiellosis in animals. Clusters of abortions in cattle by C. burnetii have not been reported previously, to our knowledge; this bacterium has been considered an opportunistic pathogen associated only with sporadic abortion in cattle. We present herein a cluster of 4 bovine abortions caused by C. burnetii in a dairy farm during a period of 2 mo and a review of the literature on C. burnetii infection in cattle.


Assuntos
Aborto Animal/microbiologia , Doenças dos Bovinos/microbiologia , Coxiella burnetii/isolamento & purificação , Complicações Infecciosas na Gravidez/veterinária , Febre Q/veterinária , Aborto Animal/epidemiologia , Animais , Bovinos , Coxiella burnetii/genética , Feminino , Feto/microbiologia , Feto/patologia , Humanos , Imuno-Histoquímica , Placenta/microbiologia , Placenta/patologia , Reação em Cadeia da Polimerase/veterinária , Gravidez , Complicações Infecciosas na Gravidez/epidemiologia , Complicações Infecciosas na Gravidez/microbiologia , Febre Q/complicações , Febre Q/epidemiologia , Febre Q/microbiologia , Uruguai/epidemiologia
11.
Am J Obstet Gynecol ; 221(2): 146.e1-146.e23, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31055031

RESUMO

BACKGROUND: Numerous reports have documented bacteria in the placental membranes and basal plate decidua in the absence of immunopathology using histologic techniques. Similarly, independent metagenomic characterizations have identified an altered taxonomic makeup in association with spontaneous preterm birth. Here we sought to corroborate these findings by localizing presumptive intact bacteria using molecular histology within the placental microanatomy. OBJECTIVE: Here we examined for microbes in term and preterm gestations using a signal-amplified 16S universal in situ hybridization probe set for bacterial rRNA, alongside traditional histologic methods of Warthin-Starry and Gram stains, as well as clinical culture methodologies. We further sought to differentiate accompanying 16S gene sequencing taxonomic profiles from germ-free (gnotobiotic) mouse and extraction and amplicon contamination controls. STUDY DESIGN: Placentas were collected from a total of 53 subjects, composed of term labored (n = 4) and unlabored cesarean deliveries (n = 22) and preterm vaginal (n = 18) and cesarean deliveries (n = 8); a placenta from a single subject with clinical and histologic evident choriomanionitis was employed as a positive control (n = 1). The preterm cohort included spontaneous preterm birth with (n = 6) and without (n = 10) preterm premature rupture of membranes, as well as medically indicated preterm births (n = 10). Placental microbes were visualized using an in situ hybridization probe set designed against highly conserved regions of the bacterial 16S ribosome, which produces an amplified stable signal using branched DNA probes. Extracted bacterial nucleic acids from these same samples were subjected to 16S rRNA metagenomic sequencing (Illumina, V4) for course taxonomic analysis, alongside environmental and kit contaminant controls. A subset of unlabored, cesarean-delivered term pregnancies were also assessed with clinical culture for readily cultivatable pathogenic microbes. RESULTS: Molecular in situ hybridization of bacterial rRNA enabled visualization and localization of low-abundance microbes after systematic high-power scanning. Despite the absence of clinical or histologic chorioamnionitis in 52 of 53 subjects, instances of 16S rRNA signal were confidently observed in 13 of 16 spontaneous preterm birth placentas, which was not significantly different from term unlabored cesarean specimens (18 of 22; P > .05). 16S rRNA signal was largely localized to the villous parenchyma and/or syncytiotrophoblast, and less commonly the chorion and the maternal intervillous space. In all term and unlabored cesarean deliveries, visualization of evident placental microbes by in situ hybridization occurred in the absence of clinical or histologic detection using conventional clinical cultivation, hematoxylin-eosin, and Gram staining. In 1 subject, appreciable villous bacteria localized to an infarction, where 16S microbial detection was confirmed by Warthin-Starry stain. In all instances, parallel sample principle coordinate analysis using Bray-Cutis distances of 16S rRNA gene sequencing data demonstrated consistent taxonomic distinction from all negative or potential contamination controls (P = .024, PERMANOVA). Classification from contaminant filtered data identified a distinct taxonomic makeup among term and preterm cohorts when compared with contaminant controls (false discovery rate <0.05). CONCLUSION: Presumptively intact placental microbes are visualized as low-abundance, low-biomass and sparse populations within the placenta regardless of gestational age and mode of delivery. Their taxonomic makeup is distinct from contamination controls. These findings further support several previously published findings, including our own, which have used metagenomics to characterize low-abundance and low-biomass microbial communities in the placenta.


Assuntos
Placenta/microbiologia , RNA Ribossômico 16S , Adulto , Código de Barras de DNA Taxonômico , Feminino , Humanos , Hibridização In Situ , Metagenômica , Microbiota , Gravidez , Nascimento Prematuro , RNA Bacteriano/análise , Análise de Sequência de RNA , Nascimento a Termo
12.
Clin Microbiol Rev ; 32(3)2019 06 19.
Artigo em Inglês | MEDLINE | ID: mdl-31043389

RESUMO

Prenatal exposure to various stressors can influence both early and later life childhood health. Microbial infection of the intrauterine environment, specifically within the placenta, has been associated with deleterious birth outcomes, such as preterm birth, as well as adverse neurological outcomes later in life. The relationships among microorganisms in the placenta, placental function, and fetal development are not well understood. Microorganisms have been associated with perinatal inflammatory responses that have the potential for disrupting fetal brain development. Microbial presence has also been associated with epigenetic modifications in the placenta, as well other tissues. Here we review research detailing the presence of microorganisms in the placenta and associations among such microorganisms, placental DNA methylation, perinatal inflammation, and neurodevelopmental outcomes.


Assuntos
Transtornos do Neurodesenvolvimento/etiologia , Doenças Placentárias/microbiologia , Placenta/microbiologia , Metilação de DNA , Feminino , Humanos , Inflamação , Doenças Placentárias/patologia , Gravidez
13.
Reprod Biol ; 19(2): 113-118, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31023521

RESUMO

Contrary to the traditional assumption of a sterile uterus, the number of studies characterizing microbial entities in the healthy upper reproductive tract (endometrial cavity, including follicular fluid and placenta) have been on the increase. Substantial data has been accumulated correlating microbial composition with fertility outcome. In this context, the presence of certain taxa was associated to an improved reproductive success. A summarization for the evidence of these molecular mechanisms through which bacteria may affect developmental processes during pregnancy is presented and discussed with special focus placed upon the immunological aspects.


Assuntos
Líquido Amniótico/microbiologia , Bactérias/isolamento & purificação , Microbiota , Placenta/microbiologia , Útero/microbiologia , Bactérias/classificação , Feminino , Humanos , Gravidez
15.
Parasitol Int ; 70: 98-101, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30822533

RESUMO

Neospora caninum, the causative agent of neosporosis, is recognised as a significant trigger of abortion and productivity losses in cattle worldwide. Current information regarding to the prevalence of N. caninum in Thailand is limited due to the limitations of detection methods and the difficulty of recovering of viable parasite. Vertical transmission is the main route of N. caninum infection in cattle. Therefore, detection of N. caninum DNA in placental tissue could be a possible means of laboratory diagnosis of neosporosis in live animals, particularly in the context of transplacental transmission. The aim of this study was to investigate the prevalence of transplacentally transmitted N. caninum infection in female beef cattle in the northern Thai province of Phayao by detection of N. caninum DNA in bovine placenta by PCR. A total of 96 bovine placentas were collected from 7 districts of Phayao. Our result indicated that overall PCR prevalence of N. caninum in cattle in this area was 36.5% varying from 16.7-50.0% between districts. The districts with the highest prevalence of infection were Muang (50.0%) and Mae Chai (44.7%). The proportion of N. caninum infection was quite high suggesting that newborn calves were at risk of congenital infection. This study provides a current snapshot of the status of bovine neosporosis in Phayao which could lead to the development of effective strategies for prevention and control this disease.


Assuntos
Doenças dos Bovinos/epidemiologia , Bovinos/microbiologia , Coccidiose/veterinária , Transmissão Vertical de Doença Infecciosa/veterinária , Placenta/microbiologia , Animais , Doenças dos Bovinos/congênito , Doenças dos Bovinos/microbiologia , Coccidiose/congênito , Coccidiose/epidemiologia , DNA Fúngico/genética , Feminino , Neospora/genética , Neospora/isolamento & purificação , Reação em Cadeia da Polimerase , Gravidez , Prevalência , Tailândia/epidemiologia
16.
Am J Obstet Gynecol ; 220(3): 267.e1-267.e39, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30832984

RESUMO

BACKGROUND: The human placenta has been traditionally viewed as sterile, and microbial invasion of this organ has been associated with adverse pregnancy outcomes. Yet, recent studies that utilized sequencing techniques reported that the human placenta at term contains a unique microbiota. These conclusions are largely based on the results derived from the sequencing of placental samples. However, such an approach carries the risk of capturing background-contaminating DNA (from DNA extraction kits, polymerase chain reaction reagents, and laboratory environments) when low microbial biomass samples are studied. OBJECTIVE: To determine whether the human placenta delivered at term in patients without labor who undergo cesarean delivery harbors a resident microbiota ("the assemblage of microorganisms present in a defined niche or environment"). STUDY DESIGN: This cross-sectional study included placentas from 29 women who had a cesarean delivery without labor at term. The study also included technical controls to account for potential background-contaminating DNA, inclusive in DNA extraction kits, polymerase chain reaction reagents, and laboratory environments. Bacterial profiles of placental tissues and background technical controls were characterized and compared with the use of bacterial culture, quantitative real-time polymerase chain reaction, 16S ribosomal RNA gene sequencing, and metagenomic surveys. RESULTS: (1) Twenty-eight of 29 placental tissues had a negative culture for microorganisms. The microorganisms retrieved by culture from the remaining sample were likely contaminants because corresponding 16S ribosomal RNA genes were not detected in the same sample. (2) Quantitative real-time polymerase chain reaction did not indicate greater abundances of bacterial 16S ribosomal RNA genes in placental tissues than in technical controls. Therefore, there was no evidence of the presence of microorganisms above background contamination from reagents in the placentas. (3) 16S ribosomal RNA gene sequencing did not reveal consistent differences in the composition or structure of bacterial profiles between placental samples and background technical controls. (4) Most of the bacterial sequences obtained from metagenomic surveys of placental tissues were from cyanobacteria, aquatic bacteria, or plant pathogens, which are microbes unlikely to populate the human placenta. Coprobacillus, which constituted 30.5% of the bacterial sequences obtained through metagenomic sequencing of placental samples, was not identified in any of the 16S ribosomal RNA gene surveys of these samples. These observations cast doubt as to whether this organism is really present in the placenta of patients at term not in labor. CONCLUSION: With the use of multiple modes of microbiologic inquiry, a resident microbiota could not be identified in human placentas delivered at term from women without labor. A consistently significant difference in the abundance and/or presence of a microbiota between placental tissue and background technical controls could not be found. All cultures of placental tissue, except 1, did not yield bacteria. Incorporating technical controls for potential sources of background-contaminating DNA for studies of low microbial biomass samples, such as the placenta, is necessary to derive reliable conclusions.


Assuntos
Microbiota , Placenta/microbiologia , Adulto , Cesárea , Estudos Transversais , Contaminação por DNA , DNA Bacteriano/análise , Feminino , Marcadores Genéticos , Humanos , Metagenômica , Microbiota/genética , Gravidez , RNA Ribossômico 16S/genética , Reação em Cadeia da Polimerase em Tempo Real , Análise de Sequência de DNA , Nascimento a Termo
17.
BMJ Open ; 9(1): e023417, 2019 01 15.
Artigo em Inglês | MEDLINE | ID: mdl-30782707

RESUMO

INTRODUCTION: Preterm birth (PTB) results from heterogeneous influences and is a major contributor to neonatal mortality and morbidity that continues to have adverse effects on infants beyond the neonatal period. This protocol describes the procedures to determine molecular signatures predictive of PTB through high-frequency sampling during pregnancy, at delivery and the postpartum period. METHODS AND ANALYSIS: Four hundred first trimester pregnant women from either Myanmar or Thailand of either Karen or Burman ethnicity, with a viable, singleton pregnancy will be enrolled in this non-interventional, prospective pregnancy birth cohort study and will be followed through to the postpartum period. Fortnightly finger prick capillary blood sampling will allow the monitoring of genome-wide transcript abundance in whole blood. Collection of stool samples and vaginal swabs each trimester, at delivery and postpartum will allow monitoring of intestinal and vaginal microbial composition. In a nested case-control analysis, perturbations of transcript abundance in capillary blood as well as longitudinal changes of the gut, vaginal and oral microbiome will be compared between mothers giving birth to preterm and matched cases giving birth to term neonates. Placenta tissue of preterm and term neonates will be used to determine bacterial colonisation as well as for the establishment of coding and non-coding RNA profiles. In addition, RNA profiles of circulating, non-coding RNA in cord blood serum will be compared with those of maternal peripheral blood serum at time of delivery. ETHICS AND DISSEMINATION: This research protocol that aims to detect perturbations in molecular trajectories preceding adverse pregnancy outcomes was approved by the ethics committee of the Faculty of Tropical Medicine, Mahidol University in Bangkok, Thailand (Ethics Reference: TMEC 15-062), the Oxford Tropical Research Ethics Committee (Ethics Reference: OxTREC: 33-15) and the local Tak Province Community Ethics Advisory Board. The results of this cooperative project will be disseminated in multiple publications staggered over time in international peer-reviewed scientific journals. TRIAL REGISTRATION NUMBER: NCT02797327; Pre-results.


Assuntos
Microbiota/fisiologia , Nascimento Prematuro/sangue , Feminino , Humanos , Placenta/microbiologia , Gravidez , Nascimento Prematuro/fisiopatologia , Estudos Prospectivos , Ultrassonografia Pré-Natal , Vagina/microbiologia
19.
Clin Perinatol ; 46(1): 29-38, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30771817

RESUMO

Necrotizing enterocolitis is a major cause of mortality and morbidity in the preterm infant population. The gut microbiome is of particular interest in research surrounding necrotizing enterocolitis, because variations in the intestinal microbiota seem to correlate with the risk of inflammation and disease. Recent advances in non-culture-based genomic sequencing have also allowed for more intricate analyses of the intestinal microbiome. Its evolution seems to be influenced by intrauterine and extrauterine factors, ranging from antenatal antibiotic exposure to type of enteral feeds. Ultimately, these alterations in the gut microbiome have the potential to result in devastating diseases like necrotizing enterocolitis.


Assuntos
Antibacterianos/uso terapêutico , Cesárea , Disbiose/microbiologia , Nutrição Enteral , Enterocolite Necrosante/microbiologia , Microbioma Gastrointestinal , Fórmulas Infantis , Leite Humano , Parto Obstétrico , Feminino , Humanos , Recém-Nascido , Recém-Nascido Prematuro , Placenta/microbiologia , Gravidez , Efeitos Tardios da Exposição Pré-Natal
20.
Arq. bras. med. vet. zootec. (Online) ; 71(1): 86-92, jan.-fev. 2019. tab, graf
Artigo em Português | LILACS, VETINDEX | ID: biblio-989387

RESUMO

A imuno-histoquímica (IHQ) é considerada uma ferramenta rápida e precisa para a identificação de protozoários, como Toxoplasma gondii, em tecidos fetais e placentários. Neste estudo foi avaliada a imunodetecção de Toxoplasma gondii em tecido placentário de cabras naturalmente infectadas. Foram coletadas e analisadas 80 amostras de placentas de cabras procedentes de único rebanho com sorologia positiva para T. gondii na técnica de ELISA. Na histopatologia, 27/80 amostras apresentaram lesões sugestivas de infecção por protozoários. Após a avaliação histopatológica, procedeu-se à realização da técnica de imuno-histoquímica, obtendo-se 85,2% (23/27) de amostras com marcação positiva. A imunodetecção ocorreu no epitélio de revestimento das vilosidades coriônicas e foi classificada de acordo com o grau de intensidade da imunomarcação. Também foi evidenciada imunomarcação no interior dos vasos sanguíneos fetais em 8,69% (2/23) das amostras. Este estudo demonstrou que a técnica de IHQ se comportou como uma ferramenta valiosa no diagnóstico da infeção por T. gondii em tecido placentário de cabras naturalmente infectadas e complementou, de forma decisiva, o diagnóstico, além de agregar maior valor aos resultados obtidos nas análises histopatológica e sorológica.(AU)


Immunohistochemistry (IHC) is considered to be a rapid and accurate tool for the identification of protozoa such as Toxoplasma gondii in fetal and placental tissues. In this study, we evaluated the immunodetection of Toxoplasma gondii in placental tissue from naturally infected goats. A total of 80 samples of goat placentas from a single herd with positive ELISA serology for T. gondii were collected and analyzed. In the histopathology, 27/80 samples presented lesions suggestive of protozoal infection. After the histopathological evaluation, the immunohistochemistry technique was performed, obtaining 85.2% (23/27) of samples with positive marking. Immunodetection occurred in the lining epithelium of the chorionic villi and was classified according to the degree of intensity of the immunostaining. Immunostaining within the fetal blood vessels was also evidenced in 8.69% (2/23) of the samples. This study demonstrated that the IHQ technique behaved as a valuable tool in the diagnosis of T. gondii infection in placental tissue of naturally infected goats completing the diagnosis in a decisive way and adding greater value to the results obtained in the histopathological and serological analysis.(AU)


Assuntos
Animais , Placenta/microbiologia , Toxoplasma/imunologia , Ruminantes/microbiologia
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