Presence Of Bacterial DNA Research Articles

Using Next-Generation Sequencing to Understand Infection Prevention in Surgical Treatment of Upper Extremity Fractures-A Prospective Cohort Study.

Postoperative fracture site infection can lead to notable patient morbidity, increase cost of care, and further contribute to healthcare disparities globally. Dogma suggests surgical blades as a vehicle for introducing bacteria into the surgical site; however, there is a paucity of literature to support this claim. This study uses advanced DNA sequencing to detect bacterial DNA on surgical blades used in upper extremity fracture surgeries. This was a prospective study, conducted at a high-volume level 1 trauma center. All acute, closed upper extremity fractures requiring surgical stabilization were consecutively enrolled in a prospective fashion. The primary end point was the presence of bacterial DNA on the surgical blade using next-generation sequencing (NGS). At the time of surgery, two blades were sterilely opened. One blade served as the control while the other was used for the initial skin incision. Two negative control blades were opened directly into a sterile container. Two positive control blades were used for skin incision through known infections. All samples were sent for NGS analysis. Forty patients were enrolled in this study. The median age was 33.5 years, and 30% were female; the median body mass index was 26.52. Humerus fractures were the most common injury (N = 17, 42.5%), followed by clavicle fractures (13, 32.5%) and radius/ulna fractures (10, 25.0%). NGS analysis revealed no contamination of test blades used for skin incision. Three control blades tested positive for bacterial DNA. Negative control blades tested negative for bacterial DNA (0/2); the positive control blades resulted positive for bacterial DNA contamination (2/2). Surgical blades used for skin incision in the upper extremity are not contaminated with bacterial DNA as analyzed by NGS. This finding challenges previous surgical dogma regarding surgical blade contamination and supports that the same surgical blade can safely be used for deeper dissection. Level II study: IRB approval-IRB#848938.

Bacterial DNA and osteoarthritis in dogs with patellar luxation and cranial cruciate ligament rupture.

The association between bacterial DNA in stifle joints, including those with cranial cruciate ligament rupture (CCLR) and medial patellar luxation (MPL), and osteoarthritis in dogs remains elusive. This study investigated the potential association between the detection of bacterial DNA and osteoarthritis in dogs using a broad-range polymerase chain reaction technique targeting the 16S ribosomal RNA gene. Synovial fluid (35 samples) and knee tissue samples (32 samples) were obtained from 35 dogs diagnosed with CCLR (n = 20; 11 males and nine females) or MPL (n = 15; five males and 10 females) who underwent a surgical operation between October 2014 and April 2015. Dogs with CCLR had a higher average osteoarthritis score than those with MPL (2.0 ± 0.9 vs. 0.5 ± 0.9; p = 0.005). Bacterial DNA was detected in the stifle joints of 60.71% of dogs with MPL. Pelomonas spp. (25.00%), Halomonas spp. (17.86%), and 5 other species (17.86%) were the most frequently identified bacteria. Bacterial DNA was detected in 41.03% of dogs with CCLR. Pelomonas spp. (15.38%), Sphingomonas spp. (10.26%), Halomonas spp. (5.13%), and 4 other species (10.26%) were the most frequently identified bacteria. No significant difference was observed in the prevalence of bacterial DNA obtained from tissue samples (46.88%) or joint fluid samples (51.43%). The presence of bacterial DNA was not associated with the type of knee injury (MPL or CCLR; p = 1.000). There was a higher prevalence of bacterial DNA in samples from dogs with moderate-to-severe osteoarthritis (94.44%) than in those with minimal osteoarthritis (41.18%), and a significant association between the presence of bacterial DNA and moderate-to-severe osteoarthritis was identified (p < 0.01). Dogs with moderate-to-severe osteoarthritis were more likely to have bacterial DNA in their stifle joints than those with no or minimal osteoarthritis. These findings provide valuable insight into the potential role of bacterial DNA in joint tissue or joint fluid and the development of osteoarthritis in dogs.

A self-powered biosensing system based on triboelectric nanogenerator for rapid bacterial DNA detection

A self-powered biosensing system based on triboelectric nanogenerator (TENG) for bacterial DNA detection is invented and a vertical contact-separation TENG was designed to provide a steady power supply to a detection bioreactor. A carboxyl-functionalized capture probe was modified on etched indium tin oxide (ITO) glass, with which target bacterial DNA could be recognized. Meanwhile, multi-walled carbon-nanotubes (CNT)-modified signal probe was designed to amplify the detection signal due to its excellent electrical conductivity. The self-powered biosensing system could measure the variation in output voltage and be highly sensible with a low detection limit at 0.084 pM of target DNA. In addition, a mathematical model based on the Freundlich isotherm was built to simulate the DNA detection process, and it was feasible to simulate the calibration equation and detection range. A portable self-powered biosensing device was made by integrating miniaturized TENG and bioreactor in a 3D-printed mold, with which the presence of bacterial DNA could be instantly indicated by LED (light-emitting diode). This study provided a useful platform for quick bacterial DNA detection and paved the way for further development of pocket diagnosis with TENG-based biosensing systems.

The potential of Rhipicephalus microplus as a vector of Ehrlichia ruminantium in West Africa

Heartwater, or cowdriosis, is a virulent tick-borne rickettsial disease of ruminants caused by Ehrlichia ruminantium, biologically transmitted by Amblyomma species (A. variegatum in West Africa). In West Africa, this bacterium was recently reported to naturally infect the invasive cattle tick, Rhipicephalus microplus (Rm) through trans-ovarian transmission from replete adult females to offspring. A ‘sheep-tick-sheep’ cycle was set up to determine whether feeding the progeny of these ticks on naïve sheep could lead to infection, and to compare clinical outcomes resulting from this transmission with those observed following infection by the natural A. variegatum (Av) vector. Using local strains of ticks (KIMINI-Rm and KIMINI-Av) and of E. ruminantium (BK242), we recorded, using the PCR technique, the presence of bacterial DNA in ticks (larvae for Av and females for Rm) engorged on sheep inoculated by BK242-infected blood. The bacterial DNA was also detected in the next stages of the lifecycle of R. microplus (eggs and larvae), and in sheep infested either by those R. microplus larvae or by A. variegatum nymphs moulted from larvae engorged on blood-inoculated sheep. Bacterial infection in these sheep was demonstrated by detecting antibodies to E. ruminantium using the MAP1-B ELISA and by isolation of the bacterium on cell culture from blood. The sequences of PCS20 gene detected in ticks and sheep were identical to that of the BK242 strain. Our results confirm that R. microplus can acquire and transmit E. ruminantium to the next stage. However, this transmission resulted in a mild subclinical disease whereas severe clinical disease was observed in sheep infested by A. variegatum infected nymphs, suggesting differences in the tick/bacteria relationship. Future studies will focus on replicating these findings with ticks of different isolates and life stages to determine if R. microplus is playing a role in the epidemiology of heartwater in West Africa. Additionally, studies will investigate whether sheep that are seropositive due to infestation by E. ruminantium-infected R. microplus are subsequently protected against heartwater. Such data will add to our understanding of the possible impact of R. microplus in areas where it has become recently established.

Human circulating bacteria and dysbiosis in non-infectious diseases.

Blood microorganisms were once thought to indicate infection. Blood in healthy people appears to be devoid of growing bacteria; nonetheless, intracellular dormant forms of bacteria have been reported previously. With breakthroughs in sequencing and bioinformatics, the presence of bacterial DNA in healthy human blood initiated the controversy of human blood microbiota (HBM). Recently, bacteria-specific DNA and culturable bacteria were found in healthy human blood. Researchers wanted to study the phenomena of a “healthy blood microbiota” by providing a thorough description of bacterially produced nucleic acids using many complementing molecular and traditional microbiological approaches. Because blood is a relatively limited and particular environment, culturability and plate count issues can be overcome using enhanced cultured procedures. However, more evidence is required to confirm that healthy human blood contains normal microbiota. Cavities, mouth and intestinal microbiota, trauma, surgery, and animal/insect bites can introduce bacteria into human blood. All these factors strengthen the concept of transient blood bacteria too. The presence of blood bacteria may be caused by temporary immunological clearance and absorption by dendritic or M cells. This review provides an extensive and comprehensive analysis that suggests that healthy blood bacteria may not be typical microbiota but transient circulatory microorganisms. In this study, we look at how contaminants (Escherichia, Shigella, Pseudomonads, etc.) from the skin, laboratory environments, and reagents can affect the interpretation of blood-derived microbial information and the relationship between the circulating bacteria and non-communicable diseases. Circulating transient bacteria may play a role in the pathogenesis of non-infectious diseases such as diabetes and CVD. Contamination-free hematological studies can aid in understanding the disease mechanisms, therapy, and biomarkers.

Plasma cfDNA predictors of established bacteraemic infection.

Introduction. Increased plasma cell-free DNA (cfDNA) has been reported for various diseases in which cell death and tissue/organ damage contribute to pathogenesis, including sepsis. Gap Statement. While several studies report a rise in plasma cfDNA in bacteraemia and sepsis, the main source of cfDNA has not been identified. Aim. In this study, we wanted to determine which of nuclear, mitochondrial or bacterial cfDNA is the major contributor to raised plasma cfDNA in hospital subjects with bloodstream infections and could therefore serve as a predictor of bacteraemic disease severity. Methodology. The total plasma concentration of double-stranded cfDNA was determined using a fluorometric assay. The presence of bacterial DNA was identified by PCR and DNA sequencing. The copy numbers of human genes, nuclear β globin and mitochondrial MTATP8, were determined by droplet digital PCR. The presence, size and concentration of apoptotic DNA from human cells were established using lab-on-a-chip technology. Results. We observed a significant difference in total plasma cfDNA from a median of 75 ng ml−1 in hospitalised subjects without bacteraemia to a median of 370 ng ml−1 (P=0.0003) in bacteraemic subjects. The copy numbers of nuclear DNA in bacteraemic also differed between a median of 1.6 copies µl−1 and 7.3 copies µl−1 (P=0.0004), respectively. In contrast, increased mitochondrial cfDNA was not specific for bacteraemic subjects, as shown by median values of 58 copies µl−1 in bacteraemic subjects, 55 copies µl−1 in other hospitalised subjects and 5.4 copies µl−1 in healthy controls. Apoptotic nucleosomal cfDNA was detected only in a subpopulation of bacteraemic subjects with documented comorbidities, consistent with elevated plasma C-reactive protein (CRP) levels in these subjects. No bacterial cfDNA was reliably detected by PCR in plasma of bacteraemic subjects over the course of infection with several bacterial pathogens. Conclusions. Our data revealed distinctive plasma cfDNA signatures in different groups of hospital subjects. The total cfDNA was significantly increased in hospital subjects with laboratory-confirmed bloodstream infections comprising nuclear and apoptotic, but not mitochondrial or bacterial cfDNAs. The apoptotic cfDNA, potentially derived from blood cells, predicted established bacteraemia. These findings deserve further investigation in different hospital settings, where cfDNA measurement could provide simple and quantifiable parameters for monitoring a disease progression.

Biofilms in Water Hoses of a Meat Processing Environment Harbor Complex Microbial Communities.

Safe and hygienic water distribution is essential for maintaining product quality and safety. It is known that biofilms alter the appearance and microbial quality of water along the distribution chain. Yet, biofilms in water hoses throughout the food processing environment have not been investigated in detail. Here, microbial communities from water hoses and other environmental sites in contact with water, in addition to the source water itself, were studied in the meat processing environment. Biofilms were present in all water hoses as determined by the presence of bacterial DNA and biofilm matrix components (carbohydrates, extracellular DNA, and proteins). The microbial community of the biofilms was dominated by Proteobacteria, represented mainly by Comamonadaceae and Pseudoxanthomonas. Moreover, genera that are associated with an intracellular lifestyle (e.g., Neochlamydia and Legionella) were present. Overall, the microbial community of biofilms was less diverse than the water microbial community, while those from the different sample sites were distinct from each other. Indeed, only a few phyla were shared between the water hose biofilm and the source water or associated environmental samples. This study provides first insights towards understanding the microbiota of water hose biofilms in the food processing environment.

Effect of delayed entry of blood culture bottles in BACTEC automated blood culture system in the context of laboratory consolidation

Delayed entry of blood culture bottles is frequent in consolidated laboratories. A retrospective study evaluated time from insertion to detection and total detection time as a function of preincubation time, and we prospectively looked for false negative results. 69,604 blood culture bottles were reviewed for preincubation time, incubation time and total detection time. Positive cultures for specific bacterial subtypes were reviewed to assess the effect of preincubation time on likelihood of detection. 492 negative blood cultures were prospectively tested by 16S RNA PCR and Staphylococcus-specific PCR for the presence of bacterial DNA. Mean preincubation time for samples collected within the city-limits was 3.94 h versus 9.49–18.89 h for other client sites. Higher preincubation times were partially mitigated by a lower incubation time, with an overall increase in total detection time. A lower odds ratio of recovery of Staphylococcus spp was identified, but not confirmed by terminal subcultures and molecular assays. Prolonged preincubation of blood cultures affects total detection time despite a reduction in incubation time. Successful centralization of microbiological services may depend upon optimization of courier routes for inoculated blood culture bottles. Our data supports consideration for an increase in suggested maximum preincubation times.

Investigation of Transovarial Transmission of Bartonella henselae in Rhipicephalus sanguineus sensu lato Ticks Using Artificial Feeding.

Bartonella henselae is a slow-growing, Gram-negative bacterium that causes cat scratch disease in humans. A transstadial transmission of the bacteria from larvae to nymphs of Rhipicephalus sanguineus sensu lato (s.l.) ticks, suspected to be a potential vector of the bacteria, has been previously demonstrated. The present study aims to investigate transovarial transmission of B. henselae from R. sanguineus s.l. adults to their instars. Adult ticks (25 males and 25 females) were fed through an artificial feeding system on B. henselae-infected goat blood for 14 days, and 300 larvae derived from the experimentally B. henselae-infected females were fed on noninfected goat blood for 7 days. Nested PCR and culture were used to detect and isolate B. henselae in ticks and blood samples. Bartonella henselae DNA was detected in midguts, salivary glands, and carcasses of the semi-engorged adults and pooled tick feces (during feeding and post-feeding periods). After the oviposition period, B. henselae DNA was detected in salivary glands of females (33.3%), but not in pooled eggs or larvae derived from the infected females. However, B. henselae DNA was detected by nested PCR from the blood sample during larval feeding, while no viable B. henselae was isolated by culture. According to our findings, following infected blood meal, B. henselae could remain in the tick midguts, move to other tissues including salivary glands, and then be shed through tick feces with limited persistency. The presence of bacterial DNA in the blood during larval feeding shows the possibility of transovarial transmission of B. henselae in R. sanguineus s.l. ticks.

Presence of bacterial DNA in thrombotic material of patients with myocardial infarction

Infectious agents have been suggested to be involved in etiopathogenesis of Acute Coronary Syndrome (ACS). However, the relationship between bacterial infection and acute myocardial infarction (AMI) has not yet been completely clarified. The objective of this study is to detect bacterial DNA in thrombotic material of patients with ACS with ST-segment elevation (STEMI) treated with Primary Percutaneous Coronary Intervention (PPCI). We studied 109 consecutive patients with STEMI, who underwent thrombus aspiration and arterial peripheral blood sampling. Testing for bacterial DNA was performed by probe-based real-time Polymerase Chain Reaction (PCR). 12 probes and primers were used for the detection of Aggregatibacteractinomycetemcomitans, Chlamydiapneumoniae, viridans group streptococci, Porphyromonasgingivalis, Fusobacteriumnucleatum, Tannarellaforsythia, Treponemadenticola, Helycobacterpylori, Mycoplasmapneumoniae, Staphylococusaureus, Prevotella intermedia and Streptococcusmutans. Thus, DNA of four species of bacteria was detected in 10 of the 109 patients studied. The most frequent species was viridans group streptococci (6 patients, 5.5%), followed by Staphylococusaureus (2 patients, 1.8%). Moreover, a patient had DNA of Porphyromonasgingivalis (0.9%); and another patient had DNA of Prevotella intermedia (0.9%). Bacterial DNA was not detected in peripheral blood of any of our patients. In conclusion, DNA of four species of endodontic and periodontal bacteria was detected in thrombotic material of 10 STEMI patients. Bacterial DNA was not detected in the peripheral blood of patients with bacterial DNA in their thrombotic material. Bacteria could be latently present in plaques and might play a role in plaque instability and thrombus formation leading to ACS.

Molecular investigation of Rickettsia spp. and Francisella tularensis in ticks from three provinces of Turkey.

Ticks are obligate hematophagous ectoparasites as well as mechanical and biological vectors of a wide variety of microbial pathogens. To date, 19 tick-borne diseases have been reported from Turkey. In this study, ticks collected from Aydın, İzmir and Şanlıurfa provinces of Turkey were identified using morphological and molecular methods. After the presence of bacterial DNA was checked, Rickettsia spp. and Francisella tularensis were investigated in bacterial DNA-positive tick specimens by PCR. Furthermore, amplicons belonging to tick specimens and positive bacterial samples were sequenced and processed for BLAST, alignment and phylogenetic analysis. As a result, seven tick species were identified: Rhipicephalus sanguineus, Rh. bursa, Rh. turanicus, Hyalomma marginatum, Hy. aegyptium, Hy. anatolicum and Haemaphysalis erinacei. Fifty-five tick specimens tested positive for bacterial DNA and among them, rickettsial DNA was found in five ticks (infection rate = 9.1%) belonging to Hy. marginatum, Hy. aegyptium, Rh. bursa and Rh. turanicus. Of the five Rickettsia-positive ticks, three contained Rickettsia aeschlimannii, one Ri. massiliae and one an unidentified Rickettsia sp. No Francisella tularensis DNA was detected. Sequence analysis of the ompB gene indicated two novel single nucleotide polymorphisms (SNP) in two different Ri. aeschlimannii strains and two novel SNPs as well as a novel insertion (GACGGT) were found in Rickettsia sp. This study indicated the presence of polymorphic Rickettsia species in ticks from Turkey.

Modulation of Placental Gene Expression in Small-for-Gestational-Age Infants.

Small-for-gestational-age (SGA) infants are fetuses that have not reached their genetically programmed growth potential. Low birth weight predisposes these infants to an increased risk of developing cardiovascular, metabolic and neurodevelopmental conditions in later life. However, our understanding of how this pathology occurs is currently incomplete. Previous research has focused on understanding the transcriptome, epigenome and bacterial signatures separately. However, we hypothesise that interactions between moderators of gene expression are critical to understanding fetal growth restriction. Through a review of the current literature, we identify that there is evidence of modulated expression/methylation of the placental genome and the presence of bacterial DNA in the placental tissue of SGA infants. We also identify that despite limited evidence of the interactions between the above results, there are promising suggestions of a relationship between bacterial signatures and placental function. This review aims to summarise the current literature concerning fetal growth from multiple avenues and propose a novel relationship between the placental transcriptome, methylome and bacterial signature that, if characterised, may be able to improve our current understanding of the placental response to stress and the aetiology of growth restriction.

Prevalence of Coxiella burnetii (Legionellales: Coxiellaceae) Infection Among Wildlife Species and the Tick Hyalomma lusitanicum (Acari: Ixodidae) in a Meso-Mediterranean Ecosystem.

Q fever is a worldwide zoonosis caused by Coxiella burnetii (Derrick) Philip. It is a major cause of abortion among sheep and may be responsible for reproductive losses in red deer in Spain. Airborne transmission is the most widespread; however, some studies suggested that ticks may play a role, but little is known about their actual involvement in the C. burnetii cycle. The aim of this study was to determine the role that Hyalomma lusitanicum (Koch) tick plays in the maintenance of this agent among wildlife in the meso-Mediterranean areas. We processed by PCR 53 swabs from wild rabbits, 21 liver samples from red deer, and 236 ticks collected at different stages. Coxiella burnetii DNA was detected in 43.40% of wild rabbits and 38.09% of red deer, supporting the hypothesis that these animals are quite likely to serve as a reservoir in the field. We also found a high prevalence of C. burnetii in ticks (55.66%). It is worth noting that 50.45% of positive ticks were collected from negative hosts, suggesting that the pathogen probably was acquired at a previous tick stage. Our results suggest transstadial transmission, and the presence of bacterial DNA in the offspring of positive female ticks is the first evidence of the transovarial transmission of C. burnetii by H. lusitanicum. Thus, this tick species seems to play an important role as a bridge of infection in the wildlife cycle, although further studies are needed to confirm vector competence.

Impact of Bacterial Translocation on Sarcopenia in Patients with Decompensated Cirrhosis.

Advanced liver disease is associated with a persistent inflammatory state, derived from abnormal bacterial translocation from the gut, which may contribute to the development of sarcopenia in cirrhosis. We aim to document the association of chronic inflammation and bacterial translocation with the presence of sarcopenia in cirrhosis. We prospectively followed cirrhotic patients aged 18–70 years with medically refractory ascites at a single tertiary care center in Toronto, Canada. The baseline data included patient demographic variables, the presence of bacterial DNA in serum/ascitic fluid, systemic inflammatory response syndrome (SIRS) status, and nutritional assessment. Thirty-one patients were enrolled, 18 (58.1%) were sarcopenic, 9 (29%) had bacterial DNA in serum and ascites fluid. The mean MELD score was 11.5 ± 4.0 (6–23). Sarcopenic and non-sarcopenic patients did not differ significantly in their baseline MELD scores, caloric intake, resting energy expenditure, the incidence of bacterial translocation, or SIRS. While sarcopenia was not linked to increased hospital admissions or death, it was strongly associated with increased episodes of acute kidney injury (3 vs. 0, p = 0.05). This pilot study did not demonstrate an association between sarcopenia and SIRS or bacterial translocation. These results should be confirmed in future larger studies, encompassing a greater number of chronic inflammation events and quantifying levels of bacterial DNA.

Diversity profiling of xenic cultures of Dientamoeba fragilis following systematic antibiotic treatment and prospects for genome sequencing.

The presence of bacterial DNA in Dientamoeba fragilis DNA extracts from culture poses a substantial challenge to sequencing the D. fragilis genome. However, elimination of bacteria from D. fragilis cultures has proven difficult in the past, presumably due to its dependence on some unknown prokaryote/s. This study explored options for removal of bacteria from D. fragilis cultures and for the generation of genome sequence data from D. fragilis. DNA was extracted from human faecal samples and xenic D. fragilis cultures. Extracts were subjected to 16S ribosomal DNA bacterial diversity profiling. Xenic D. fragilis cultures were then subject to antibiotic treatment regimens that systematically removed bacterial species depending on their membrane structure (Gram-positive or Gram-negative) and aerobic requirements. The impact of these treatments on cultures was assessed by 16S amplicon sequencing. Prior to antibiotic treatment, the cultures were dominated by Gram-negative bacteria. Addition of meropenem to cultures eliminated anaerobic Gram-negative bacteria, but it also led to protozoan death after 5 days incubation. The seeding of meropenem resistant Klebsiella pneumoniae strain KPC-2 into cultures before treatment by meropenem prevented death of D. fragilis cells beyond this 5 day period, suggesting that one or more species of Gram-negative bacteria may be an essential nutritional requirement for D. fragilis. Gram-positive cells were completely eliminated using vancomycin without affecting trophozoite growth. Finally, this study shows that genome sequencing of D. fragilis is feasible following bacterial elimination from cultures as the result of the major advances occurring in bioinformatics. We provide evidence on this fact by successfully sequencing the D. fragilis 28S large ribosomal DNA subunit gene using culture-derived DNA.

Investigation of the skin microbiome

The traditional way of finding out which bacteria are present in the skin is to take a sample using a swab. One limitation of this is that swab samples are taken from the skin surface only, whereas bacteria may be present in the deeper layers of the skin as well. Researchers from Sweden, Ireland and the UK looked at the difference between the results of swabs and actual biopsies (tissue samples) of the skin. One swab sample and one 2 mm punch biopsy from the same site were taken from each of 16 patients who were having lesions on their trunk or limbs excised (surgically removed) anyway. The swab used was moistened first, and the biopsy was taken down to the fat layer. A technique called 16S rRNA gene sequencing was used on the biopsy samples to search for the presence of bacteria. This is a very sensitive technique that can pick up the presence of bacterial DNA even if bacteria cannot be cultured (grown). The biopsies showed a significant increase in the groups of bacteria called Clostridiales and Bacteroidetes. Clostridiales do not need oxygen to live, and so might be expected to be found in the deeper layers of the skin. Other bacteria such as the common Staphylococcus aureus, on the other hand, were more abundant in the swab samples. Knowing more accurately which bacteria live in the skin is important because these bacteria can trigger the immune system responses that are important in skin diseases like atopic dermatitis, psoriasis and hidradenitis suppurativa.

Human placenta has no microbiome but can contain potential pathogens.

We sought to determine whether preeclampsia, delivery of a small for gestational age infant or spontaneous preterm birth 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 acquisition of bacteria during labour and delivery or contamination of laboratory reagents with bacterial DNA. The exception was Streptococcus agalactiae (Group B Streptococcus), where non-contaminant signals were detected in ~5% of samples collected prior to 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.

A Survey of Bacterial Pathogens Detected in Feces and Wool in Small Ruminants (Pilot Study)

Sheep feces can carry a high concentration of pathogenic and non-pathogenic bacteria, which potentially may contaminate wool as well as the shearers or wool manipulators through direct contact. A pilot study was carried out to determine the presence of bacterial DNA in feces and the degree of bacterial contamination in wool in two species of ruminants. Fourteen 2-month old lambs and 14 kids (7 male and 7 female), uncastrated, no twins, with their mothers, were randomly selected at weaning from a free flock grazing on naturalized pasture of Los Rios region, Chile. Fecal and wool samples were taken once and analyzed for genomic DNA of Salmonella typhimurium containing the virulence plasmid spv, Eschrichia coli serotype O157, Clostridium perfringens type C containing α toxin and Mycobacterium avium sp paratuberculosis containing the IS900 insertion element. The results showed that lamb and kids feces had higher contents of bacterial DNA for E. coli O157 and SalmT than lamb wool, although only one lamb showed these two bacteria on its wool. The bacterial species influenced the DNA expression for 16S in both, feces (P=0.05) and wool (P=0.0006) and for E. coli O157 and SalmT only in feces (P<0.0001). The sex was associated with E. coli detection in lambs feces (P<0.0007) and in kids feces (P<0.05). The values obtained for MAP IS900 and Cpa DNA contents, considering both species and sex, were undetectable. In conclusion, lamb and kids feces should potentially contaminate wool especially by Eschrichia coli O157 and Salmonella typhimurium, representing a potential health risk and public health concern, especially for shearers and wool handlers.

Treatment with non-selective beta-blockers affects the systemic inflammatory response to bacterial DNA in patients with cirrhosis.

Background & AimsThe use of non‐selective beta‐blockers has been associated with lower rates of infection and reduced infection‐associated morbidity in patients with cirrhosis. However, it is unknown if these drugs modify the systemic inflammatory response to circulating bacterial DNA.MethodsSixty‐three patients with cirrhosis were included during an episode of decompensation by ascites. Thirty of those patients were on beta‐blockers. Blood samples were obtained after each patient had been in the supine position for at least 30 minutes in a quiet atmosphere. Bacterial DNA, serum cytokines, nitric oxide, and LPS were determined. Phagocytic and oxidative burst activities were determined in polymorphonuclear cells from the patients.ResultsThe detection rate of bacterial DNA in the blood was the same (33%) for patients not treated and treated with non‐selective beta‐blockers. Patients naive to non‐selective beta‐blockers showed significantly higher serum levels of IL6, IFN‐gamma and IL10 in response to the presence of bacterial DNA. Patients treated with non‐selective beta‐blockers showed higher basal inflammatory activity that did not change with the presence of bacterial DNA. Monocytes and granulocytes from patients treated with non‐selective beta‐blockers showed a significantly increased phagocytic capacity in the presence of bacterial DNA.ConclusionsIn patients with cirrhosis, chronic treatment with beta‐blockers is associated with a higher unstimulated production of serum cytokines and an increased phagocytic activity in the presence of bacterial DNA.

Prevotella strains and lactamic resistance gene distribution in different oral environments of children with pulp necrosis.

To determine the presence of Prevotella species, the cfxA/cfxA2, blaZ and blaTEM genes associated with resistance to lactamic agents in different oral niches of children with pulp necrosis. Children with pulp necrosis in primary teeth had samples of saliva, supragingival, pulp chamber and root canal biofilms collected and tested for Prevotella species (P.intermedia, P.nigrescens, P.tannerae) and for beta-lactam resistance genes (cfxA/cfxA2, blaZ and blaTEM). The presence of bacterial DNA was examined through PCR, with a specific primer directed to the 16S rRNA gene. Specific primers were used to detect the Prevotella species and beta-lactam resistance genes. The chi-square test was used to analyse associations between the presence of bacteria and clinical variables. The Cochran's Q test was used to assess whether the proportion of gene detection is the same between different sites. Thirty-two teeth were sampled from 27 patients with a mean age of 5.5years (±1.76). The total detection rate of Prevotella strains was 29.1%, 25%, 21.8% and 32.29% in saliva, supragingival, pulp chamber and root canal samples, respectively. P.nigrescens was the most commonly detected species in all oral niches. The previous use of antibiotics was associated with detection of P.nigrescens in saliva (P=0.03). Pain was associated with the presence of P.nigrescens (P=0.04) and P.tannerae (P= 0.01) in pulp chamber biofilm. blaTEM was detected in the four oral niches, being more frequent (23.8%) in supragingival biofilm (Cochran's Q test, P=0.04). The presence of P.intermedia in SB and PC was associated with the detection of blaTEM in saliva (P=0.04). The cfxA/cfxA2 and blaZ genes were not detected in any of the four oral niches. The oral cavity of children with pulp necrosis had a variable distribution of Prevotella strains in different niches. Saliva, supragingival biofilm, pulp chamber and root canals of primary teeth with necrotic pulps can harbour resistance genes to beta-lactams agents.