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(1) Background: The surveillance of SARS-CoV-2 RNA in urban wastewaters allows one to monitor the presence of the virus in a population, including asymptomatic and symptomatic individuals, capturing the real circulation of this pathogen. The aim of this study was to evaluate the performance of different pre-analytical and analytical methods for identifying the presence of SARS-CoV-2 in untreated municipal wastewaters samples by conducting an inter-laboratory proficiency test. (2) Methods: three methods of concentration, namely, (A) Dextran and PEG-6000 two-phase separation, (B) PEG-8000 precipitation without a chloroform purification step and (C) PEG-8000 precipitation with a chloroform purification step were combined with three different protocols of RNA extraction by using commercial kits and were tested by using two primers/probe sets in three different master mixes. (3) Results: PEG-8000 precipitation without chloroform treatment showed the best performance in the SARS-CoV-2 recovery; no major differences were observed among the protocol of RNA extraction and the one-step real-time RT-PCR master mix kits. The highest analytic sensitivity was observed by using primers/probe sets targeting the N1/N3 fragments of SARS-CoV-2. (4) Conclusions: PEG-8000 precipitation in combination with real-time RT-PCR targeting the N gene (two fragments) was the best performing workflow for the detection of SARS-CoV-2 RNA in municipal wastewaters.

During the first period of the SARS-CoV-2 pandemic, the lack of specific therapeutic treatments led to the provisional use of a number of drugs, with a continuous review of health protocols when new scientific evidence emerged. The management of this emergency sanitary situation could not take care of the possible indirect adverse effects on the environment, such as the release of a large amount of pharmaceuticals from wastewater treatment plants. The massive use of drugs, which were never used so widely until then, implied new risks for the aquatic environment. In this study, a suspect screening approach using Liquid Chromatography-High Resolution Mass Spectrometry techniques, allowed us to survey the presence of pharmaceuticals used for COVID-19 treatment in three WWTPs of Lombardy region, where the first European cluster of SARS-CoV-2 cases was detected. Starting from a list of sixty-three suspect compounds used against COVID-19 (including some metabolites and transformation products), six compounds were fully identified and monitored together with other target analytes, mainly pharmaceuticals of common use. A monthly monitoring campaign was conducted in a WWTP from April to December 2020 and the temporal trends of some anti-COVID-19 drugs were positively correlated with those of COVID-19 cases and deaths. The comparison of the average emission loads among the three WWTPs evidenced that the highest loads of hydroxychloroquine, azithromycin and ciprofloxacin were measured in the WWTP which received the sewages from a hospital specializing in the treatment of COVID-19 patients. The monitoring of the receiving water bodies evidenced the presence of eight compounds of high ecological concern, whose risk was assessed in terms of toxicity and the possibility of inducing antibiotic and viral resistance. The results clearly showed that the enhanced, but not completely justified, use of ciprofloxacin and azithromycin represented a risk for antibiotic resistance in the aquatic ecosystems.

The use of computational structural models that include geometrical non-linearity in many application cases may require high reliability in prediction of displacements. Nevertheless, large differences up to 60% on maximum total displacement have been found among results of static large-deformation analyses performed by means of the major commercial software packages in a simple benchmark study with linear material properties. In order to investigate the causes of such disagreement, the present work compares different finite element formulations including well-established stress update schemes. The various formulations are tested, and results are compared in three test cases. Rodriguez stress update algorithms have shown the best performance among methods reported in literature. Finally, the cause of the large differences found in the predictions of commercial codes is identified. It is linked to the energetic inconsistency of some stress update methods in the simulation of extension/compression loading conditions. Such inaccuracy is reproduced analytically by formulating and integrating the corresponding inconsistent constitutive equations. The identified problem is very important for designers, as it affects almost all the static simulations, which are the most common type of large-deformation analyses and usually involve extension/compression loading.

Wastewater-based epidemiology has been proposed to monitor the diffusion and trend of SARS-CoV-2 pandemic. In the present study, raw and treated samples from three wastewater treatment plants, and two river samples characterized the Milano Metropolitan Area, Italy, were surveyed for SARS-CoV-2 RNA positivity to real time PCR and infectiveness. Moreover, whole genome sequencing and phylogenetic analysis of isolated strains was performed. Raw wastewater samples resulted positive to PCR amplification, while treated water samples were always negative (four and two samples, respectively, sampled in two dates). Moreover, the rate of positivity in raw wastewater samples decreased after eight days, in congruence with the epidemiological trend estimated for the interested provinces. Virus infectiveness was always not significant, indicating the effectiveness of wastewater treatments, or the natural decay of viral vitality, which implied the absence of significant risk of infection from wastewaters. Samples from receiving rivers (two sites, sampled in the same dates as wastewaters) showed in some cases a positivity to PCR amplification, probably due to non-treated discharges, or the combined sewage overflows. Nevertheless, also for rivers vitality was negligible, indicating the absence of sanitary risks. Phylogenetic analysis of genome indicated that the isolated virus belongs to the most spread strain present in Europe and similar to another strain found in Lombardy.

The aim of the present research is to characterize both experimentally and numerically journal bearings with low radial clearances for rotors in small-scale applications (e.g., microgas turbines); their diameter is in the order of ten millimetres, leading to very small dimensional clearances when the typical relative ones (order of 1/1000) are employed; investigating this particular class of journal bearings under static and dynamic loading conditions represents something unexplored. To this goal, a suitable test rig was designed and the performance of its bearings was investigated under steady load. For the sake of comparison, numerical simulations of the lubrication were also performed by means of a simplified model. The original test rig adopted is a commercial rotor kit (RK), but substantial modifications were carried out in order to allow significant measurements. Indeed, the relative radial clearance of RK4 RK bearings is about 2/100, while it is around 1/1000 in industrial bearings. Therefore, the same original RK bearings are employed in this new test rig, but a new shaft was designed to reduce their original clearance. The new custom shaft allows to study bearing behaviour for different clearances, since it is equipped with interchangeable journals. Experimental data obtained by this test rig are then compared with further results of more sophisticated simulations. They were carried out by means of an in-house developed finite element (FEM) code, suitable for thermoelasto-hydrodynamic (TEHD) analysis of journal bearings both in static and dynamic conditions. In this paper, bearing static performances are studied to assess the reliability of the experimental journal location predictions by comparing them with the ones coming from already validated numerical codes. Such comparisons are presented both for large and small clearance bearings of original and modified RKs, respectively. Good agreement is found only for the modified RK equipped with small clearance bearings (relative radial clearance 8/1000), as expected. In comparison with two-dimensional lubrication analysis, three-dimensional simulation improves prediction of journal location and correlation with experimental results.

The lagoons of the Po River delta are potentially exposed to complex mixtures of contaminants, nevertheless, there is a substantial lack of information about the biological effects of these contaminants in the Po delta lagoons. These environments are highly dynamic and the interactions between chemical and environmental stressors could prevent the proper identification of biological effects and their causes. In this study, we aimed to disentangle such interactions focusing on Manila clams, previously exposed to six lagoons of the Po delta, adopting three complementary tools: a) the detailed description via modelling techniques of lagoon dynamics for salinity and water temperature; b) the response sensitivity of a number of target genes (ahr, cyp4, ρ-gst, σ-gst, hsp22, hsp70, hsp90, ikb, dbh, ach, cat, Mn-sod, Cu/Zn-sod, cyp-a, flp, grx, TrxP) investigated in clam digestive glands by Real Time PCR; and c) the relevance of DNA adducts determined in clams as markers of exposure to genotoxic chemicals. The lagoons showed specific dynamics, and two of them (Marinetta and Canarin) could induce osmotic stress. A group of genes (ahr, cyp4, Mn-sod, σ-gst, hsp-22, cyp-a, TrxP) seemed to be associated with overall lagoon characteristics as may be described by salinity and its variations. Lagoon modelling and a second group of genes (hsp70, hsp90, cat, ikb, ach, grx, Cu/Zn-sod) also suggested that moderate increases of river discharge may imply worse exposure conditions. Oxidative stress seemed to be associated with such events but it was slightly evident also under normal exposure conditions. DNA adduct formation was mainly associated with overwhelmed antioxidant defences (e.g. low Cu/Zn-sod) or seemingly with their lack of response in due time. In Po delta lagoons, Manila clam can be affected by chemical and environmental factors which can contribute to induce oxidative stress, DNA adduct formation and, ultimately, to affect clam condition and health.

Space exploration is demanding longer lasting human missions and water resupply from Earth will become increasingly unrealistic. In a near future, the spacecraft water monitoring systems will require technological advances to promptly identify and counteract contingent events of waterborne microbial contamination, posing health risks to astronauts with lowered immune responsiveness. The search for bio-analytical approaches, alternative to those applied on Earth by cultivation-dependent methods, is pushed by the compelling need to limit waste disposal and avoid microbial regrowth from analytical carryovers. Prospective technologies will be selected only if first validated in a flight-like environment, by following basic principles, advantages, and limitations beyond their current applications on Earth. Starting from the water monitoring activities applied on the International Space Station, we provide a critical overview of the nucleic acid amplification-based approaches (i.e., loop-mediated isothermal amplification, quantitative PCR, and high-throughput sequencing) and early-warning methods for total microbial load assessments (i.e., ATP-metry, flow cytometry), already used at a high readiness level aboard crewed space vehicles. Our findings suggest that the forthcoming space applications of mature technologies will be necessarily bounded by a compromise between analytical performances (e.g., speed to results, identification depth, reproducibility, multiparametricity) and detrimental technical requirements (e.g., reagent usage, waste production, operator skills, crew time). As space exploration progresses toward extended missions to Moon and Mars, miniaturized systems that also minimize crew involvement in their end-to-end operation are likely applicable on the long-term and suitable for the in-flight water and microbiological research.

The aim of the present work is to design a test rig suited to investigate the dynamic interaction between rotor and hydrodynamic journal bearings in micro gas turbines (microGT), i.e. with reference to small bearings (diameter in the order of ten millimeters). Particularly, the device is capable of measuring the journal location. Therefore, the journal motion due to rotor vibrations can be displayed, in order to assess performance as well as stiffness and damping of the bearings. The new test rig is based on Bently Nevada Rotor Kit (RK), but substantial modifications are carried out. Indeed, the relative radial clearance of the original RK bearings is about 2/100, while it is in the order of 1/1000 in industrial bearings. Therefore, the same RK bearings are employed in the new test rig, but a new shaft has been designed in order to reduce the original clearance. The new shaft enables us to study the bearing behaviour for different clearances, as it is equipped with interchangeable journals. The experimental data yielded by the new test rig are compared with numerical results. These are obtained by means of a suitable finite element (FEM) code developed by our research group. It allows the Thermo Elasto-HydroDynamic (TEHD) analysis of the bearing in static and dynamic conditions. In the present paper, bearing static performances are analysed in order to assess the reliability of the journal location predictions by comparing numerical and experimental results. Such comparisons are presented for both large and small clearance bearings of original and modified RK, respectively. Good agreement is found only for the modified RK equipped with small clearance bearings (relative radial clearance equal to 8/1000). Nevertheless, rotor alignment is quite difficult with small clearance bearings and a completely new test rig is designed for future experiments.

The presence of SARS-CoV-2 in raw wastewaters has been demonstrated in many countries affected by this pandemic. Nevertheless, virus presence and infectivity in treated wastewaters, but also in the receiving water bodies are still poorly investigated. In this study, raw and treated samples from three wastewater treatment plants, and three river samples within the Milano Metropolitan Area, Italy, were surveyed for SARS-CoV-2 RNA detection by means of real time RT-PCR and infectivity test on culture cells. SARS-CoV-2 RNA was detected in raw, but not in treated wastewaters (four and two samples, respectively, sampled in two dates). The isolated virus genome was sequenced, and belonged to the strain most spread in Europe and similar to another found in the same region. RNA presence in raw wastewater samples decreased after eight days, probably following the epidemiological trend estimated for the area. Virus infectivity was always null, indicating the natural decay of viral pathogenicity in time from emission. Samples from receiving rivers (three sites, sampled in the same dates as wastewaters) showed in some cases a positivity to real time RT-PCR, probably due to non-treated, or inefficiently treated discharges, or to the combined sewage overflows. Nevertheless, also for rivers infectivity was null. Risks for public health should be limited, although a precautionary approach to risk assessment is here advocated, giving the preliminary nature of the presented data., Graphical abstract Unlabelled Image, Highlights • SARS-CoV-2 RNA presence and infectivity in wastewaters and receptors was assessed. • Viral RNA was detectable in the inflow but not in the outflow wastewaters. • Viral RNA was present in receptors due to sewage overflows or inefficient treatment. • SARS-CoV-2 infectivity was null both in wastewaters and receptors. • A precautionary approach in the assessment of contagious risk is advocated.

Sediment toxicity plays a fundamental role in the health of inland fish communities; however, the assessment of the hazard potential of contaminated sediments is not a common objective in environmental diagnostics or remediation. This study examined the potential of transcriptional endpoints investigated in zebrafish (Danio rerio) exposed to riverbed sediments in ecotoxicity testing. Embryo-larval 10-day tests were conducted on sediment samples collected from five sites (one upstream and four downstream of the city of Milan) along a polluted tributary of the Po River, the Lambro River. Sediment chemistry showed a progressive downstream deterioration in river quality, so that the final sampling site showed up to eight times higher concentrations of, for example, triclosan, galaxolide, PAH, PCB, BPA, Ni, and Pb, compared with the uppermost site. The embryo/larval tests showed widespread toxicity although the middle river sections evidenced worse effects, as evidenced by delayed embryo development, hatching rate, larval survival, and growth. At the mRNA transcript level, the genes encoding biotransformation enzymes (cyp1a, gst, ugt) showed increasing upregulations after exposure to sediment from further downstream sites. The genes involved in antioxidant responses (sod, gpx) suggested that more critical conditions may be present at downstream sites, but even upstream of Milan there seemed to be some level of oxidative stress. Indirect evidences of potential apoptotic activity (bcl2/bax < 1) in turn suggested the possibility of genotoxic effects. The genes encoding for estrogen receptors (er alpha, er beta 1, er beta 2) showed exposure to (xeno)estrogens with a progressive increase after exposure to sediments from downstream sites, paralleled by a corresponding downregulation of the ar gene, likely related to antiandrogenic compounds. Multiple levels of thyroid disruption were also evident particularly in downstream zebrafish, as for thyroid growth (nkx2.1), hormone synthesis and transport (tg, ttr, d2), and signal transduction (tr alpha, tr beta). The inhibition of the igf2 gene reasonably reflected larval growth inhibitions. Although none of the sediment chemicals could singly explain fish responses, principal component analysis suggested a good correlation between gene transcripts and the overall trend of contamination. Thus, the combined impacts from known and unknown covarying chemicals were proposed as the most probable explanation of fish responses. In summary, transcriptional endpoints applied to zebrafish embryo/larval test can provide sensitive, comprehensive, and timeliness information which may greatly enable the assessment of the hazard potential of sediments to fish, complementing morphological endpoints and being potentially predictive of longer studies.

A multigenerational test with Chironomus riparius was performed to assess long-term effects on life-traits of exposure to selected perfluoroalkyl compounds: perfluorooctane sulfonate (PFOS), perfluorooctanoic acid (PFOA), and perfluorobutane sulfonate (PFBS). These persistent contaminants are widespread in aquatic ecosystems at low concentrations, possibly exerting long-term toxicity. Larvae of C. riparius of a native population were exposed for 10 generations to 10 μg/L nominal concentrations of PFOS, PFOA, and PFBS, comparable with the maximum values found in European rivers. All treatments showed reduced growth at most/several generations. No effects on survival, development, and reproduction were found. A final tolerance-induction test was performed exposing the pre-exposed experimental cohorts to 100 µg/L PFOS and 150 µg/L PFOA for a whole life cycle. Factorial analysis of variance revealed no difference between treatments (i.e., PFOS vs PFOA), indicating no induced tolerance. Instead, organisms pre-exposed to PFBS were the most stressed, followed by those pre-exposed to PFOA and PFOS, with earlier emergence and reduced adult weight. The results may be related to general stress and genetic erosion induced by long-term laboratory culture, but also to long-term toxicant exposure. However, no effects at the population level (population growth rate) were proved, and thus a toxicity risk in real ecosystems at the tested concentrations seems unlikely. Environ Toxicol Chem 2019;00:1-12. © 2019 SETAC.

Scleractinian corals (i.e. hard corals) play a fundamental role in building and maintaining coral reefs, one of the most diverse ecosystems on Earth. Nevertheless, their phylogenies remain largely unresolved and little is known about dispersal and survival of their planktonic larval phase. The small subunit ribosomal RNA (SSU rRNA) is a commonly used gene for DNA barcoding in several metazoans, and small variable regions of SSU rRNA are widely adopted as barcode marker to investigate marine plankton community structure worldwide. Here, we provide a large sequence data set of the complete SSU rRNA gene from 298 specimens, representing all known extant reef coral families and a total of 106 genera. The secondary structure was extremely conserved within the order with few exceptions due to insertions or deletions occurring in the variable regions. Remarkable differences in SSU rRNA length and base composition were detected between and within acroporids (Acropora, Montipora, Isopora and Alveopora) compared to other corals. The V4 and V9 regions seem to be promising barcode loci because variation at commonly used barcode primer binding sites was extremely low, while their levels of divergence allowed families and genera to be distinguished. A time-calibrated phylogeny of Scleractinia is provided, and mutation rate heterogeneity is demonstrated across main lineages. The use of this data set as a valuable reference for investigating aspects of ecology, biology, molecular taxonomy and evolution of scleractinian corals is discussed.

a b s t r a c t A multigeneration toxicity test on Chironomus riparius was performed with the aim of investigating the evolutionary consequences of exposure to perfluoralkyl substances (perfluorooctane sulfonic acid, PFOS; perfluorooctanoic acid, PFOA; perfluorobutane sulfonate, PFBS). Six-hundred larvae were bred per treatment and per generation until emergence and egg deposition under a nominal concentration of 10 g/L of contaminants. Newborn larvae were used to start the next generation. Evolution of genetic variability was evaluated along a total of 10 consecutive generations based on 5 microsatellite loci. Analysis of life-history traits (survival, sex ratio and reproduction) was also carried out. Rapid genetic variability reduction was observed in all treatments, including controls, across generations due to the test conditions. Nevertheless, an increased mutation rate determined a stronger conservation of genetic variability in PFOS and, at minor extent, in PFBS exposed populations compared to controls. No significant effects were induced by exposure to PFOA. Direct mutagenicity or induced stress conditions may be at the base of increased mutation rate, indicating the potential risk of mutational load caused by exposure to PFOS and PFBS. The test provided the opportunity to evaluate the use of approximate Bayesian computation (ABC) and coalescent approaches in evolutionary ecotoxicology. A weak performance was evidenced for ABC, either in terms of bias or dispersion of effective population sizes and of estimates of mutation rate. On the contrary, coalescent simulations proved the sensitivity of traditional genetic endpoints (i.e. heterozygosity and number of alleles) to the alteration of mutation rate, but not to erosion of genetic effective size. © 2014 Elsevier B.V. All rights reserved.

A new scleractinian coral species, Pachyseris inattesa sp. n., is described from the Red Sea. Despite a superficial resemblance with some species in the agariciid genus Leptoseris with which it has been previously confused, P. inattesa sp. n. has micro-morphological characters typical of the genus Pachyseris. This genus, once part of the Agariciidae, is comprised of five extant species and is widely distributed throughout the tropical Indo-Pacific. It is currently incertae sedis as a result of recent molecular analysis and appears to be closely related to the Euphylliidae. A molecular phylogenetic reconstruction including P. inattesa sp.n., the genus type species P. rugosa, and P. speciosa, all present in the Red Sea, was performed using the mitochondrial intergenic spacer between COI and 16S-rRNA. The results confirm that P. inattesa sp. n. is a monophyletic lineage closely related to the other Pachyseris species examined.

Purpose – The purpose of this paper was to conceive a fast method to verify design and performance of elliptical pocket journal bearings. Design/methodology/approach – The computer-aided verification of pocket journal bearings is performed by means of a suitable finite element analysis method. Findings – The results of sample analyses indicate that the machining tolerances are very influential on elliptical pocket bearing performances, and they must be included among the input data. Originality/value – Although elliptical pocket bearings are widely used in turbomachineries, the influence of their design on performance has not been specifically investigated. A lot of works about tribological models are available, but few of them focus on their application to bearing design at the industrial level.

Composite sediment samples were collected from 18 deposition areas located in seven lagoons of the Po Delta. Samples were analysed for contents of legacy (DDT, PCB) and priority organic pollutants (AP, PBDE), and the sediments showed, with some exceptions, low/moderate levels of contamination. It seemed evident that the Po River has a clear role in controlling the input and distribution of chemicals to the lagoons, generally resulting in an overall north-south pattern of increasing contamination, as well as some contaminant contents exceeding quality guidelines or threshold values (NP, PBDE, PCB). Interestingly, more negative δ13C values were observed in the lagoons exposed to freshwater inflows, with higher concentrations of DDTs in their sediments. Regarding potential toxic effects, different sex ratios were observed in Manila clams (Ruditapes philippinarum) previously exposed for 3 months to six lagoons of the Po Delta, and these ratios ranged from a marked male to female prevalence. Notably, the male clam percentage was inversely correlated with sedimentary PCB and partially correlated with PBDE, bioaccumulated PCB, PBDE and to a lesser extent APs and DDTs. A delay in gonad development was also present in the clams most exposed to sedimentary APs and in general to inland waters. For the first time, this study evidenced endocrine disruption in Manila clams exposed to the Po Delta lagoons that was apparently the result of exposure to major contaminant classes. These ecotoxicological results need to be further investigated, in light of the many other endocrine disrupters recently identified in Po lagoons.

A comprehensive analytical screening has been applied for the first time to surficial sediment samples collected from seven lagoons of the Po River Delta. This screening has been used to gain further insights into the exposure conditions potentially present in the Po lagoons. Despite a certain degree of variability was observable within the lagoons, this study confirmed the general tendency that the northernmost Caleri lagoon is less contaminated than the other lagoons of the delta, which conversely are more exposed to river waters and particularly to the Po River. This screening identified 23 suspect and 8 target contaminants in the investigated lagoons. The target contaminants were quantified by means of authentic analytical standards, and showed concentration values ranging from 0.01 ng/g for irbesartan, to 80 ng/g for DEET, respectively. The surveyed molecules included pharmaceuticals, industrial chemicals, personal care products, pesticides as well as illicit substances mainly originating from inland waters and the anthropic activities of their (sub)basins. Notably, many of these chemicals, such as flame retardants (TPhP, TCEP, TCPP), phthalates (DnBP, DiBP, DnOP), synthetic hormones (norethisterone, trenbolone) and several parabens (methyl-, ethyl-, butylparaben) are endocrine disruptors and have been detected in all Po Delta lagoons. This study showed that the lagoons of the Po Delta are exposed to a multitude of chemicals substances whose multiple interactions are very likely. As suggested by other studies conducted in this area, the disrupting activities of chemicals capable to interfere with the endocrine system of aquatic organisms should be thoroughly investigated.

In this study, we characterized the gene expression responses of the Padanian barbel (Barbus plebejus), a native benthivorous cyprinid with a very compromised presence within the fish community of the River Po. Barbel juveniles were exposed in the laboratory to two river sediments reflecting an upstream/downstream gradient of increasing contamination and collected from one of the most anthropized tributaries of the River Po. After 7 months of exposure, hepatic transcriptional changes that were diagnostic of sediment exposure were assessed. We investigated a set of 24 genes involved in xenobiotic biotransformation (cyp1a, gst?, ugt), antioxidant defense (gpx, sod, cat, hsp70), trace metal exposure (mt-I, mt-II), DNA repair (xpa, xpc), apoptosis (bax, casp3), growth (igf2), and steroid (er?, er?1, er?2, ar, vtg) and thyroid (dio1, dio2, tr?, tr?, nis) hormone signaling pathways. In a consistent overall picture, the results showed that long-term sediment exposure mainly increased the levels of mRNAs encoding proteins involved in xenobiotic metabolism, oxidative stress defense, repair of DNA damage and activation of the apoptotic process. Transcript up-regulation of three receptor genes (er?2, ar, tr?), likely representing compensatory responses to antagonistic/toxic effects, was also observed, confirming the exposure to disruptors of the reproductive and thyroidal axes. In contrast to expectations, a few genes showed no response (e.g., casp3) or even downregulation (vtg), further suggesting that the timing of exposure/assessment, potential compensatory effects or post-transcriptional modifications interact to modify the gene expression profiles, particularly during exposure to mixtures of contaminants.

Background: In 2015 a new device for the collection of mediastinal fluid from patients with deep sternal wound infection (DSWI) in the presence of negative-pressure wound therapy (NPWT) became available. The present study was designed to evaluate whether changing sample collection devices increased micro-organism detection in patients undergoing NPWT. Methods: During 2013-2014, 207 samples were collected and cultured from NPWT patients (n = 23) to demonstrate the presence of DSWI using reticulated polyurethane sponge culture, a swab, and blood culture. In 2015, a new collection device was introduced for specimen collection. A total of 357 samples (n = 17) were collected using the ESwab(TM) (Copan, Murrieta, CA) for deep and superficial wound sample collection. In addition, blood culture devices were used for collecting mediastinal fluid aspirated directly from the wound and biologic fluid obtained from the NPWT device. Fisher exact test was performed to test the rate of independence rate of micro-organism identification using the NPWT sponge device and taking blood culture results as a reference for micro-organism identification. Results: After the introduction of the new collection device in our hospital, an overall increase in the detection of micro-organisms (46.7%) was reported. During 2013-2014 our traditional microbiologic collection method did not detect a pathogen in 30.4% of patients. During 2015, the new sample collection approach, direct from the NPWT device, improved micro-organism detection by 10.4% and reduced DSWIs with undetected pathogens to 17.6% (p < 0.01). Conclusions: As a result of proficiency gained in the last year, the most representative specimen in wound infection was represented by mediastinal fluid collected directly from the wound and the NPWT device. Given the correlation between the blood culture of micro-organisms detected using the ESwab device from the wound, mediastinal drainage, and drainage from the NPWT device, we can assume that the NPWT device may replace the other biologic sampling devices.