Bacterial Plate Counts Research Articles

ASSESSMENT OF PAINT-PIGMENT DEGRADING MICROORGANISMS FROM PAINT INDUSTRIES EFFLUENT-CONTAMINATED SITES IN ABA, SOUTH-EAST NIGERIA

The potential of microorganisms isolated from paint industry effluent-polluted soils to degrade the constituents of the effluents and contaminant removal efficiency after the remediation period were evaluated. Samples were collected from six different paint industries in Aba. Samples from the various paint industries reveals sample F showed the highest bacterial plate counts of 6.21±0.20 x 104cfu/g while samples B and D had the least bacterial counts of 1.30±0.20 x 104cfu/g and 1.30±0.31 x 104cfu/g respectively. The fungal counts of the polluted soils indicates sample D as having the highest count of 2.30±0.24 x 104cfu/g while sample A had the least count of 1.00±0.30 x104cfu/g. Total Paint Utilizing Fungal (TPUF) counts ranges from 1.23±0.02 x 104cfu/g to 3.45±0.10 x 104cfu/g and showed a significant difference between sample C and the control (p<0.05). The bacterial isolates from the contaminated soil include Pseudomonas spp., Escherichia coli, Citrobacter spp., Klebsiella spp., Proteus spp., Enterobacter spp., Providencia rettgeri, Shigella flexneri, Staphylococcus spp., Streptococcus spp. and Enterococcus spp. while the fungal isolates include Saccharomyces cerevisiae, Rhodotorula species, Rhizopus species, Aspergillus niger, Aspergillus flavus and Penicillum notatum. The counts of actual paint utilizing fungi were relatively lower than the heterotrophic groups. The paint degrading microorganisms isolated from the paint industry effluent polluted soils include paint effluent bacterial degraders: Pseudomonas spp. (33.3%), Escherichia coli(33.3%),, Citrobacter spp. (25%), Klebsiella spp.(25%), Proteus spp.(25%), Enterobacter spp.(12.5%). and Providencia rettgeri(12.5%), While the fungal paint effluent degraders include Geotrichum candidum(33.3%), Pichia kudriavzevii(33.3%), Fusarium sp.(25%), Aspergillus niger(25%), Penicillium funiculosum(25%) and Mucor sp.(12.5%). Geotrichum candidum strain GAD1 (MN63874.1) and Pichia kudriavzevii strain (MK811096.1) showed highest degrees of growth indicated by high turbidities during the screening test. This study showed that majority of the Indigenous organisms has the ability to utilize paints as their sole carbon source hence are the major bioremediating agents in the sampling areas.

Detection of antibiotic-producing bacteria from soil samples in parts of Wudil local government area Kano State, Nigeria

The emergence of new diseases and multiple-antibiotic resistance pathogens that resist the action of clinically used antibiotics have increased the needs of discovering new antibiotics. However, soil served as the most important target for most researchers in their attempt of discovering new antibiotics that have pharmaceutical values. The aim of this study was to detect the antibiotic-producing bacteria from soil samples in parts of Wudil local government of Kano state using standard procedure. The color ranged from light brown to dark brown. The texture ranged from clay loam, clay and loam. Sample C was found to have the highest mean pH and electrical conductivity of 7.46 and 108.90μs/cm, respectively, while sample B had the least mean values of 6.95 and 73.53μs/cm, respectively. The highest mean bulk devsity and particle density was sample B with 1.60g/cm3 and 1.87g/cm3, respectively, while sample C had the least mean values of 1.48g/cm3 and 1.71g/cm3, respectively. Sample C was found to have the highest mean moisture content and porosity with 17.93% and 15.57%, respectively, while sample A had the least mean values with 10.61% and 10.74%, respectively. Four bacterial spp. were identified based on morphological description, gram’s reaction and biochemical tests, and were identified as Bacillus spp., Micrococcus spp., Pseudosomonas spp. and Proteus spp. Sample C was found to have the highest mean total bacterial viable plate count of 19.9×105cfu/g, while sample B had the least mean value of 4.1×105cfu/g. Bacillus spp. was found to have the highest percentage occurrence with 21(47%), while Proteus spp. had the least value with 4(9%). E. coli had the highest and least susceptibility to metabolites produced by Bacillus spp. and Pseudosomonas spp. with inhibition zones of (17 and 0mm) respectively. While the control (Erythromycin 250mg/ml) produced the inhibition zones of (27 and 21mm) against E. coli and S. aureus respectively. It was concluded that soil samples in Lajawa, Kausani, Wudil Sabon gari and Wudil Tsohon gari of Wudil local government contained the bacteria with potentiality of producing antimicrobial substance and recommended the use of molecular techniques for further characterization.

Construction of multifunctional micro-patterned PALNMA/PDADMAC/PEGDA hydrogel and intelligently responsive antibacterial coating HA/BBR on Mg alloy surface for orthopedic application

In recent years, magnesium alloys (MgA) have been reckoned as the most promising material of biomedical importance on account of its excellent degradable properties and mechanical properties mimicking natural bone tissues. However, MgA are prone to rapid corrosion under physiological conditions, causing toxicity around the neighboring tissues. In addition, they are susceptible to bacterial colonization, a detrimental factor for medical causes. In this study, antibacterial material coated hydrogel-based micro-patterns were developed on MgA to achieve long-term antibacterial, antifouling, osteogenic, and cell-compatible properties. First, the Mg(OH)2 nanosheet coating was prepared on the surface of MgA as a physical barrier to prevent the corrosion of MgA. Then the hydrogel micropatterns of poly(alendronate sodium methacrylate)/poly(dimethyldiallylammonium chloride)/poly(ethylene glycol) diacrylate (PALNMA/PDADMAC/PEGDA) of different sizes were constructed on the surface of the Mg(OH)2 coating using the photomask method. Finally, an intelligently responsive antibacterial material hyaluronic acid/berberine (HA/BBR) was coated on MgA-Mg(OH)2-PALNMA/PDADMAC/PEGDA patterns via layer-by-layer self-assembly. The excellent antifouling performance of the samples is attributed to the topological structure of the pattern. Interestingly, as the pattern size of PALNMA/PDADMAC/PEGDA decreases, the antibacterial, antifouling, and cell compatibility properties of the samples gradually improve. UV–Vis spectra and bacterial plate count indicate that HA/BBR coating provide a pH and hyaluronidase (HAase) dual-responsive surface to kill the attached bacteria quickly. Finally, the in vitro experiments demonstrate excellent blood compatibility, cell compatibility and osteogenic properties of the modified MgA samples. Therefore, the intelligent multifunctional assembly of MgA presented here has a promising future in the field of metal implant materials.

Prophylactic Knockdown of the miR-183/96/182 Cluster Ameliorates Pseudomonas aeruginosa-Induced Keratitis.

PurposePreviously, we demonstrated that miR-183/96/182 cluster (miR-183C) knockout mice exhibit decreased severity of Pseudomonas aeruginosa (PA)-induced keratitis. This study tests the hypothesis that prophylactic knockdown of miR-183C ameliorates PA keratitis indicative of a therapeutic potential.MethodsEight-week-old miR-183C wild-type and C57BL/6J inbred mice were used. Locked nucleic acid–modified anti-miR-183C or negative control oligoribonucleotides with scrambled sequences (NC ORNs) were injected subconjunctivally 1 day before and then topically applied once daily for 5 days post-infection (dpi) (strain 19660). Corneal disease was graded at 1, 3, and 5 dpi. Corneas were harvested for RT-PCR, ELISA, immunofluorescence (IF), myeloperoxidase and plate count assays, and flow cytometry. Corneal nerve density was evaluated in flatmounted corneas by IF staining with anti-β-III tubulin antibody.ResultsAnti-miR-183C downregulated miR-183C in the cornea. It resulted in an increase in IL-1β at 1 dpi, which was decreased at 5 dpi; fewer polymorphonuclear leukocytes (PMNs) at 5 dpi; lower viable bacterial plate count at both 1 and 5 dpi; increased percentages of MHCII+ macrophages (Mϕ) and dendritic cells (DCs), consistent with enhanced activation/maturation; and decreased severity of PA keratitis. Anti-miR-183C treatment in the cornea of naïve mice resulted in a transient reduction of corneal nerve density, which was fully recovered one week after the last anti-miR application. miR-183C targets repulsive axon-guidance receptor molecule Neuropilin 1, which may mediate the effect of anti-miR-183C on corneal nerve regression.ConclusionsProphylactic miR-183C knockdown is protective against PA keratitis through its regulation of innate immunity, corneal innervation, and neuroimmune interactions.

Microbiological Analysis of Liquid Lipstick Products Used by Makeup Artists

AbstractLipstick is a non-sterile pharmaceutical product that is used for make-up on the lips. Liquid lipstick used by makeup artists may have a high level of microbial contamination because they are applied to different clients in turn. This study aims to determine the microbiological quality of liquid lipstick used by makeup artists based on the result of the Bacterial Total Plate Count (TPC), Yeast and Mold Count (YMC), and the presence of pathogenic microbes. Three samples of liquid lipstick were taken from three different makeup artists and one sample was a new liquid lipstick product. The Bacterial TPC test was carried out on TSA media and the YMC test was carried out on SDA with the spread plate technique. The presence of microbial pathogens P. aeruginosa, S. aureus, and C. albicans were carried out on selective media. The results of bacterial TPC was 9.0 × 103 to 3.0 × 104 CFU/mL, while the YMC was 1.1 × 103 to 3.1 × 103 CFU/mL. The pathogenic microbes detected were S. aureus in 2 samples and C. albicans in 1 sample. The bacterial TPC and YMC values exceed the contamination limit based on BPOM Regulation No. 12/2019.

IMPACT OF ULTRAVIOLET RADIATION ON BACTERIAL GROWTH OF KN95 MASK

Pandemic SARS CoV-2 is currently spreading around the world. Preventive measures to be implemented include using Personal Protective Equipment (PPE) especially mask which lead to a lack of mask supply. Strategy is proposed by decontaminating it so that it can be used repeatedly. This research was conducted to compare the effectiveness of UV radiation disinfection in different duration on the KN95 mask. This research was conducted by decontaminating of KN95 mask using UV radiation with radiation time of 15, 30, and 45 minutes. The mask was swab before and after the radiation. Examination for the growth of pathogen bacterial colonies and total plate count method was done to determine the effectivity. It was found the number of germs before and after has decreased significantly radiation at 30 (P = 0.036) and 45 minutes (P = 0.037). It also can be effective in decontaminating pathogen bacterial as the number colony of Staphylococcus aureus has decreased significantly in 30 minutes (P=0.034) and 45 minutes exposure (P= 0.037). The results of this study indicate that UV radiation for 30 and 45 minutes can be effective in decontaminating pathogen bacterial

Rapid Methodologies for Assessing Pseudomonas syringae pv. actinidiae Colonization and Effector-Mediated Hypersensitive Response in Kiwifruit.

The infection of Pseudomonas syringae pv. actinidiae in kiwifruit is currently assessed by numerous methodologies, each with their own limitations. Most studies are based on either a laborious method of growth quantification of the pathogen or qualitative assessments by visual scoring following stem or cutting inoculation. Additionally, when assessing for resistance against specific pathogen effectors, confounding interactions between multiple genes in the pathogen can make mapping resistance phenotypes nearly impossible. Here, we present robust alternative methods to quantify pathogen load based on rapid bacterial DNA quantification by PCR, the use of Pseudomonas fluorescens, and a transient reporter eclipse assay for assessing resistance conferred by isolated bacterial avirulence genes. These assays compare well with bacterial plate counts to assess bacterial colonization as a result of plant resistance activation. The DNA-based quantification, when coupled with the P. fluorescens and reporter eclipse assays to independently identify bacterial avirulence genes, is rapid, highly reproducible, and scalable for high-throughput screens of multiple cultivars or genotypes. Application of these methodologies will allow rapid and high-throughput identification of resistant cultivars and the bacterial avirulence genes they recognize, facilitating resistance gene discovery for plant breeding programs.[Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Unraveling the riverine antibiotic resistome: The downstream fate of anthropogenic inputs

River networks are one of the main routes by which the public could be exposed to environmental sources of antibiotic resistance, that may be introduced e.g. via treated wastewater. In this study, we applied a comprehensive integrated analysis encompassing mass-flow concepts, chemistry, bacterial plate counts, resistance gene quantification and shotgun metagenomics to track the fate of the resistome (collective antibiotic resistance genes (ARGs) in a microbial community) of treated wastewater in two Swiss rivers at the kilometer scale. The levels of certain ARGs and the class 1 integron integrase gene (intI1) commonly associated with anthropogenic sources of ARGs decreased quickly over short distances (2-2.5 km) downstream of wastewater discharge points. Mass-flow analysis based on conservative tracers suggested this decrease was attributable mainly to dilution but ARG loadings frequently also decreased (e.g., 55.0-98.5 % for ermB and tetW) over the longest studied distances (6.8 and 13.7 km downstream). Metagenomic analysis confirmed that ARG of wastewater-origin did not persist in rivers after 5 ~ 6.8 km downstream distance. sul1 and intI1 levels and loadings were more variable and even increased sharply at 5 ~ 6.8 km downstream distance on one occasion. While input from agriculture and in-situ positive selection pressure for organisms carrying ARGs cannot be excluded, in-system growth of biomass is a more probable explanation. The potential for direct human exposure to the resistome of wastewater-origin thus appeared to typically abate rapidly in the studied rivers. However, the riverine aquatic resistome was also dynamic, as evidenced by the increase of certain gene markers downstream, without obvious sources of anthropogenic contamination. This study provides new insight into drivers of riverine resistomes and pinpoints key monitoring targets indicative of where human sources and exposures are likely to be most acute.

Assessment of biochemical profile among patients of Microbiological Quality Assessment of Bakery Products Available in Lahore, Pakistan

Bakery items are a vital element of a well-balanced diet, and supermarket shelves now stock a wide range of them. Bakery items are the most popular ready-to-eat foods (REF).Objective:To assess the microbial quality of bakery foods (Bread, Cake and patties) and check the total plate count (TPC), bacterial and yeast count Methods:The analysis of the microbial quality of bakery products was determined for three different bakery products (bread, cake and patties) at ten different locations collected from a renowned bakery in Lahore, Pakistan. The samples were assessed under complete hygienic conditions by standardized plate count (TPC) and MPN method. Micro-organisms (Coliform, Fecal Coliform and Yeast) were lately confirmed by their morphological characteristics and biochemical tests Results: Among the three commodities, bread was found to be highly contaminated by micro-organisms and the total plate count recorded was 4.7 x 105cfu g-1collected from sector I9 which is not consumable. Whereas Patties showed the number of TPC as 1.6 x 105 cfu g-1also collected from sector I9. Cake was revealed as least infected bakery product i.e., 5.7 x 104 cfu g-1. The above results showed that sector I9 is highly contaminated and a threat for health. Bacterial plate count (Coliform and Fecal Coliform count) was highly measured in bread as 460 MPN g-1 taken from sector I5 where as it remains under the permissible limit determined by WHO, 1994 in remaining sectors Conclusions:In conclusion the bakery food is contaminated by the use of un-hygienic water, miss handled during the process after baking i.e., slicing, serving and packaging. To avoid a possible epidemic of food borne illness, competent authorities should conduct a stricter and more stringent inspection of REFs sold to clients in bakeries.

In situ Fabrication of Nano ZnO/BCM Biocomposite Based on MA Modified Bacterial Cellulose Membrane for Antibacterial and Wound Healing.

BackgroundDeveloping an ideal wound dressing that meets the multiple demands of safe and practical, good biocompatibility, superior mechanical property and excellent antibacterial activity is highly desirable for wound healing. Bacterial cellulose (BC) is one of such promising class of biopolymers since it can control wound exudates and can provide moist environment to a wound resulting in better wound healing. However, the lack of antibacterial activity has limited its application.Methods and ResultsWe prepared a flexible dressing based on a bacterial cellulose membrane and then modified it by chemical crosslinking to prepare in situ synthesis of nZnO/BCM via a facile and eco-friendly approach. Scanning electron microscopy (SEM) results indicated that nZnO/BCM membranes were characterized by an ideal porous structure (pore size: 30~ 90 μm), forming a unique string-beaded morphology. The average water vapor transmission of nZnO/BCM was 2856.60 g/m2/day, which improved the moist environment of nZnO/BCM. ATR-FITR further confirmed the stepwise deposition of nano-zinc oxide. Tensile testing indicated that our nanocomposites were flexible, comfortable and resilient. Bacterial suspension assay and plate counting methods demonstrated that 5wt. % nZnO/BCM possessed excellent antibacterial activity against S.aureus and E. coli, while MTT assay demonstrated that they had no measurable cytotoxicity toward mammalian cells. Moreover, skin irritation test and histocompatibility examination supported that 5wt. % nZnO/BCM had no stimulation to skin and had acceptable biocompatibility with little infiltration of the inflammatory cells. Finally, by using a bacteria-infected (S. aureus and E. coli) murine wound model, we found that nZnO/BCM could prevent in vivo bacterial infections and promote wound healing via accelerating the re-epithelialization and wound contraction, and these membranes had no obvious toxicity toward normal tissues.ConclusionTherefore, the constructed nZnO/BCM has great potential for biomedical applications as an efficient antibacterial wound dressing.

Quantification, Variability Assessment of Bacterial Pollution and Public Health Hazards Linked to Users of Automated Teller Machines in Ekpoma, Edo State-Nigeria

Raising number of mortals has used automated teller machines (ATMs) over the years, but little is known about their microbial colonization status. Based on this premise, we examined eight out of the nine commercial bank’s ATM in Ekpoma environs for quantification, variability assessment of the bacterial cross contamination and the likely health hazards linked to the users of such cash dispensing machines. A total number of sixteen (16) samples were acquired from eight different commercial banks ATM keypads and screens within the study area, using sterile swab sticks immersed in sterile normal saline. Samples were subsequently transferred to the laboratory section of the Department of Microbiology, Faculty of Life Sciences, Ambrose Alli University, Ekpoma for analyses using standard microbiological procedures for isolation, quantification and identification between the period of September to October 2019. Results showed that the total aerobic bacteria count, Staphylococcus aureus and coliforms counts on both the ATM keypads and Screens ranged from 9.0 × 103±2.65 cfu/m2 to 1.92 × 104±3.61 cfu/m2, 3.5 × 103±1.73 cfu/m2 to 9.8 × 103±4.58 cfu/m2 and 4.8 × 103±1.00 cfu/m2 to 1.08 × 104±2.00 cfu/m2 respectively. One-way Anova depicted a no significant difference (P ˃ 0.05) in the total aerobic bacterial plate count, Escherichia coli and Staphylococcus aureus counts in the various banks’ ATM keypads and screens in respectively of the facilities placement and the number of users. The bacteria implicated in this study were Escherichia coli, Bacillus spp, Klebsiella pnuemoniae, Staphylococcus aureus, Pseudomonas aeruginosa, Proteus spp and Coagulase negative Staphylococcus spp. The findings from this study reveals that high volume of bacterial pollution were detected on ATM hardware user interface which invariably necessitated the need for adequate personal hygiene by both the users and custodians of such machines with a view to reducing the likely hood of spreading contagious agents.

Wound dressing from polyvinyl alcohol/chitosan electrospun fiber membrane loaded with OH-CATH30 nanoparticles

Novel nanomaterials have been developed for antimicrobial and wound healing applications. Here, we report the preparation of a polyvinyl alcohol/chitosan (PVA/CS) nanofiber with carboxymethyl chitosan nanoparticles (CMCS-OH30 NPs) encapsulating the antibacterial peptide OH-CATH30 (OH-30). The PVA/CS nanofibers containing OH-30 NPs (NP-30-NFs) obtained via electrospinning could achieve a secondary embedded OH-30. The effect of NP-30-NFs on the release of OH-30 was investigated through high-performance liquid chromatography. The antibacterial activities of NP-30-NFs against Escherichia coli and Staphylococcus aureus were studied by bacterial plate counting. NP-30-NFs containing different concentrations of NPs were applied to mouse skin wounds to determine their effectiveness in promoting wound healing. Results showed that NP-30-NFs exhibited antibacterial properties and promoted skin wound healing.

Assessment of bacterial pathogens on edible macroalgae in coastal waters

In the Northeast USA, the aquaculture of macroalgae is a rapidly growing industry. Within this region, there are no established regulations for farm siting or methods of pathogen detection on macroalgae cultivated or harvested for human consumption. Bacterial pathogens from natural and anthropogenic sources may persist in coastal waters and can potentially contaminate macroalgae. During the winter growing season, sugar kelp Saccharina latissima and adjacent water were sampled from three sites of kelp aquaculture located in adjacent bays of ME, USA. Membrane filtration onto selective media detected Escherichia coli, Vibrio parahaemolyticus, and Vibrio alginolyticus in kelp and water samples at all sites, however plate counts were very low. The foodborne pathogens Salmonella enterica ser. Typhimurium, V. parahaemolyticus, and enterohemorrhagic E. coli O157:H7 were detected on enriched kelp samples from 83%, 78%, and 56% of sampling events, respectively, using molecular methods. Even with low bacterial levels, this frequency of detection confirms the risk of foodborne pathogens present on kelp and recommends the development of best management practices to control microbial growth during kelp harvest and processing. Bacterial plate counts from kelp samples often varied from those of water, indicating the importance of sampling the kelp directly, and that the association between bacterial pathogens on kelp and in the surrounding water should be further investigated. This study provides the first food safety assessment of sea vegetables in this region with the goal of providing data to enable the expansion of its industry.

Infection Risk Reduction Program on Pathogens in High School and Collegiate Athletic Training Rooms.

Background:Athletic training rooms have a high prevalence of bacteria, including multidrug-resistant organisms, increasing the risk for both local and systematic infections in athletes. There are limited data outlining formal protocols or standardized programs to reduce bacterial and viral burden in training rooms as a means of decreasing infection rate at the collegiate and high school levels.Hypothesis:Adaptation of a hygiene protocol would lead to a reduction in bacterial and viral pathogen counts in athletic training rooms.Study Design:Cohort study.Level of Evidence:Level 3.Methods:Two high school and 2 collegiate athletic training rooms were studied over the course of the 2017-2018 academic year. A 3-phase protocol, including introduction of disinfectant products followed by student-athlete and athletic trainer education, was implemented at the 4 schools. Multiple surfaces in the athletic training rooms were swabbed at 4 time points throughout the investigation. Bacterial and viral burden from swabs were analyzed for overall bacterial aerobic plate count (APC), bacterial adenosine triphosphate activity, influenza viral load, and multidrug-resistant organisms such as methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococcus (VRE).Results:Overall bacterial load, as measured by APC, was reduced by 94.7% (95% CI, 72.6-99.0; P = 0.003) over the course of the investigation after protocol implementation. MRSA and VRE were found on 24% of surfaces prior to intervention and were reduced to 0% by the end of the study. Influenza was initially detected on 25% of surfaces, with no detection after intervention. No cases of athletic training room–acquired infections were reported during the study period.Conclusion:A uniform infection control protocol was effective in reducing bacterial and viral burden, including multidrug-resistant organisms, when implemented in the athletic training rooms of 2 high schools and 2 colleges.Clinical Relevance:A standardized infection control protocol can be utilized in athletic training rooms to reduce bacterial and viral burden.

Amperometric genosensor for culture independent bacterial count

Bacterial plate count for general assessment of water quality requires lengthy bacterial culturing. We report here a new DNA induced current genosensor for culture independent total bacteria determination. Since the amount of bacterial DNA is correlated to the number of bacteria, the genosensor measures the amount of bacterial DNA to determine bacterial count. The approach relies on bacteria lysis to release DNA which can react with molybdate to form redox molybdophosphate and measured electrochemically. Analysis of E. coli and S. aureus demonstrated that the DNA generated current is highly correlated with the level of bacteria lysis which was confirmed by spectrometric measurement. Culture independent measurement of S. aureus bacterial load suggests limit of detection is 21.9 CFU/mL, with linear range from 3 × 102 to 3 × 107 CFU/mL and correlation coefficient of 0.992. For E. coli analysis, the detection limit is 25.1 CFU/mL with the same linear range. The use of electrochemical microbial DNA quantitation for culture independent bacterial count is a new approach, the genosensor measurement is rapid (within 1 h) and has potential use for analysis of broad-spectrum bacteria for various applications.

Impact of a Quality Improvement Infection Risk Reduction Program on Pathogen Presence in High School and Collegiate Athletic Training Room Facilities

Objectives:Athletic training rooms have a high prevalence of bacteria, including multidrug resistant organisms, increasing the risk for both local and systematic infections in athletes. There is limited data outlining formal protocols or standardized programs to reduce the bacterial and viral burden in training rooms as a means of decreasing infection rate at the collegiate and high school levels. The purpose of this study is to implement established hygiene principles to high school and collegiate athletic training rooms to reduce bacterial and viral burden using a quality improvement infection control program. The authors hypothesize that implementation of such a program will reduce bacterial and viral counts in athletic training rooms over the course of one academic year.Methods:Two high school and two college athletic training rooms were studied over the course of one academic year. A three-phase protocol, including introduction of disinfectant products, implementation of posters and checklists, and student-athlete and athletic trainer education was implemented at the four schools. Multiple surfaces in athletic training rooms were swabbed at four time points throughout the investigation, corresponding with each phase of protocol implementation (Image 1). Bacterial and viral burden from swabs were analyzed for overall bacterial aerobic plate count (APC), influenza viral load and the presence of multidrug resistant organisms such methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin resistant enterococcus (VRE).Results:Overall bacterial load, as measured by APC, was reduced by 94.7% (95% CI [72.6, 99.0], p = 0.003) over the course of the investigation following protocol implementation. Image 2 demonstrates the downward trend in bacterial load averaged across all surfaces sampled over the course of the study. MRSA and VRE were found on 24% of surfaces prior to intervention and were reduced to 0% by the end of the study. Influenza was initially detected on 25% of surfaces with no detection following intervention. No cases of athletic training room acquired infections were reported during the study period.Conclusion:A uniform infection control protocol implemented in the athletic training room of four secondary schools was effective in reducing bacterial and viral burden, including multidrug resistant organisms, over the course of one academic year. Future investigations will focus on expanding this protocol to a larger number of schools with broader demographics.

Musca domestica Cecropin (Mdc) Alleviates Salmonella typhimurium-Induced Colonic Mucosal Barrier Impairment: Associating With Inflammatory and Oxidative Stress Response, Tight Junction as Well as Intestinal Flora.

Salmonella typhimurium, a Gram-negative food-borne pathogen, induces impairment in intestinal mucosal barrier function frequently. The injury is related to many factors such as inflammation, oxidative stress, tight junctions and flora changes in the host intestine. Musca domestica cecropin (Mdc), a novel antimicrobial peptide containing 40 amino acids, has potential antibacterial, anti-inflammatory, and immunological functions. It remains unclear exactly whether and how Mdc reduces colonic mucosal barrier damage caused by S. typhimurium. Twenty four 6-week-old male mice were divided into four groups: normal group, control group (S. typhimurium-challenged), Mdc group, and ceftriaxone sodium group (Cs group). HE staining and transmission electron microscopy (TEM) were performed to observe the morphology of the colon tissues. Bacterial load of S. typhimurium in colon, liver and spleen were determined by bacterial plate counting. Inflammatory factors were detected by enzyme linked immunosorbent assay (ELISA). Oxidative stress levels in the colon tissues were also analyzed. Immunofluorescence analysis, RT-PCR, and Western blot were carried out to examine the levels of tight junction and inflammatory proteins. The intestinal microbiota composition was assessed via 16s rDNA sequencing. We successfully built and evaluated an S. typhimurium-infection model in mice. Morphology and microcosmic change of the colon tissues confirmed the protective qualities of Mdc. Mdc could inhibit colonic inflammation and oxidative stress. Tight junctions were improved significantly after Mdc administration. Interestingly, Mdc ameliorated intestinal flora imbalance, which may be related to the improvement of tight junction. Our results shed a new light on protective effects and mechanism of the antimicrobial peptide Mdc on colonic mucosal barrier damage caused by S. typhimurium infection. Mdc is expected to be an important candidate for S. typhimurium infection treatment.

Microbiomes in Ground Water and Alternative Irrigation Water, and Spinach Microbiomes Impacted by Irrigation with Different Types of Water

Irrigation water, particularly if applied overhead, could be an important source of bacterial contamination to fresh produce. The colonization, survival, and proliferation of exogenous bacterial pathogens can be strongly influenced by the produce microbiota. In this study, spinach grown in an organic field was irrigated with ground water and potential alternative irrigation water including reclaimed wastewater, and urban runoff water, over a period of 2 weeks. Water and spinach samples were collected before and after irrigation for bacterial plate count, qPCR, and community profiling using 16S rDNA high-throughput sequencing analyses. The average bacterial population densities on spinach (6.50 ± 0.04 log CFU/g, 7.40 ± 0.10 log 16S copies/g) were significantly higher than those in irrigation water (3.61 ± 0.12 log CFU/ml, 4.94 ± 0.13 log 16S copies/ml). The composition and relative abundance of spinach microbiomes varied with different types of irrigation waters; however, the most abundant microbial taxa on spinach were not significantly affected by the irrigation with different types of water. Shigella, Salmonella, Listeria, Campylobacter spp., and pathogenic Escherichia coli were not detected in this study. This study provides information on the microbial ecology of diverse bacterial communities on spinach surface after irrigation by different types of water, which can benefit future studies on the interaction of microbes on produce, and the prevention of foodborne pathogens and plant disease.

NLRP12 promotes host resistance against Pseudomonas aeruginosa keratitis inflammatory responses through the negative regulation of NF-κB signaling.

To investigate the role of NLRP12 in regulating Pseudomonas aeruginosa (P. aeruginosa) keratitis. Real-Time-PCR and Western blot were performed to measure the NLRP12 level in corneas and bone marrow-derived macrophages (BMDMs) of C57BL/6 (B6) mice. B6 mice received a subconjunctival injection of lentivirus expressing active NLRP12 (NLRP12-lentivirus) or Ctl-lentivirus (as control), followed by infection of P. aeruginosa. The clinical score, slit lamp and bacterial plate count of mice were evaluated. In addition, myeloperoxidase (MPO) was detected to assess the infiltration of polymorphonuclear neutrophil (PMN). Cytokine levels were measured by Real Time-PCR and ELISA. Meanwhile, the bacterial burden was also evaluated. The activation of NF-κB signaling was determined by pIκBα/IκBα levels based on Western blot and NF-κB-dependent Luciferase activity on the basis of Luciferase assays using 293T cells. NLRP12 mRNA and protein levels were decreased in B6 corneas and BMDMs after P. aeruginosa infection. The over-expression of NLRP12 in B6 corneas significantly ameliorated the severity of corneal disease, bacterial burden, PMN infiltration and pro-inflammatory cytokine expression. In vitro analysis demonstrated that the up-regulation of NLRP12 suppressed pro-inflammatory cytokine production and enhanced bacterial clearance in RAW264.7 cells. The protein levels of pIκBα and IκBα were significantly decreased after NLRP12-lentivirus treatment compared with that of Ctl-lentivirus. NF-κB-dependent Luciferase activity was potently inhibited by NLRP12 infected with P. aeruginosa or cotransfected with the downstream signaling molecules including IKKα and IKKβ in 293T cells. NLRP12 decreases the severity of P. aeruginosa keratitis, reduces corneal inflammation and bacterial burden through the down-regulation of the NF-κB signaling pathway.

Comparative study on the petroleum crude oil degradation potential of microbes from petroleum-contaminated soil and non-contaminated soil

Comparative study on the petroleum crude oil degradation potential of microbes from petroleum-contaminated soil and non-contaminated soil was performed in the present paper. Three samples were employed to conduct the evaluation, of which two are from the petroleum-contaminated soil of Yanchang oil field, and one is from non-contaminated soil of Northwest University Campus, Xi’an. The bacterial plate counts in sample No. 1 of the petroleum-contaminated soil were 6.0 × 102 and the bacterial plate counts in sample No. 2 of the petroleum-contaminated soil were 4.0 × 103, while the bacterial plate counts in sample No. 3 of non-petroleum-contaminated were 6.0 × 104. Petroleum-degrading microbes were isolated from Bushnell Haas mineral salt medium and 16 bacterial strains and two fungal strains from PCS sample No. 1, five bacterial strains and one fungal strain from PCS sample No. 2, and 17 bacterial strains and one fungal strain from garden soil, respectively. For primary screening of potential petroleum-degrading microbes, 2, 6-dichlorophenol indophenol test was carried out and nine bacterial strains from PCS No. 1 and four bacterial strains from garden soil showed the potential petroleum-degrading activity. The petroleum degradation percentage of mixture of 13 bacteria was 47.57%, and it was higher than that of four fungi and the combined mixture (13 bacteria and four fungi). Study showed that the microbes isolated from non-contaminated soil had the good petroleum degradation potential.