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For the past 30 years, the number of people infected with causative agents of ehrlichiosis, Rocky Mountain spotted fever, and spotted fever group rickettiosis (SFGR) has increased in Oklahoma. However, there is a lack of data on pathogen prevalence within urban environments. To assess the prevalence of tick-borne pathogens in different environments, 434 Amblyomma americanum (lone star) ticks were collected from the environment in two parks in Edmond, Oklahoma. The presence of Ehrlichia spp. and spotted fever group (SFG) Rickettsia spp. was determined using quantitative real-time polymerase chain reaction (qPCR). 33.6% (146/434) of the A. americanum ticks were positive for Rickettsia amblyommatis and 15.2% (66/434) were positive for Ehrlichia chaffeensis. No ticks were positive for other SFG Rickettsiae (R. rickettsii, R. parkeri) or other Ehrlichiae (E. ewingii, and Panola Mountain Ehrlichia). These studies provide increased understanding of the potential risk for encountering tick-borne pathogens in urban environments.

For the past 30 years, the number of people infected with causative agents of ehrlichiosis, Rocky Mountain spotted fever, and spotted fever group rickettiosis (SFGR) has increased in Oklahoma. However, there is a lack of data on pathogen prevalence within urban environments. To assess the prevalence of tick-borne pathogens in different environments, 434 Amblyomma americanum (lone star) ticks were collected from the environment in two parks in Edmond, Oklahoma. The presence of Ehrlichia spp. and spotted fever group (SFG) Rickettsia spp. was determined using quantitative real-time polymerase chain reaction (qPCR). 33.6% (146/434) of the A. americanum ticks were positive for Rickettsia amblyommatis and 15.2% (66/434) were positive for Ehrlichia chaffeensis. No ticks were positive for other SFG Rickettsiae (R. rickettsii, R. parkeri) or other Ehrlichiae (E. ewingii, and Panola Mountain Ehrlichia). These studies provide increased understanding of the potential risk for encountering tick-borne pathogens in urban environments.

Infections from antibiotic resistant Staphylococcus aureus and Pseudomonas aeruginosa are a serious threat because reduced antibiotic efficacy complicates treatment decisions and prolongs the disease state in many patients. To expand the arsenal of treatments against antimicrobial resistant (AMR) pathogens, 600-Da branched polyethylenimine (BPEI) can overcome antibiotic resistance mechanisms and potentiate β-lactam antibiotics against Gram-positive bacteria. BPEI binds cell wall teichoic acids and disables resistance factors from penicillin binding proteins PBP2a and PBP4. The present study describes a new mechanism of action for BPEI potentiation of antibiotics generally regarded as agents effective against Gram-positive pathogens but not Gram-negative bacteria. 600-Da BPEI is able to reduce the barriers to drug influx and facilitate the uptake of a non-β-lactam co-drug, erythromycin, that targets the intracellular machinery. Also, BPEI can suppress production of the cytokine interleukin IL-8 by human epithelial keratinocytes. This enables BPEI to function as a broad-spectrum antibiotic potentiator which expands the opportunities to improve drug design, antibiotic development, and therapeutic approaches against pathogenic bacteria, especially for wound care.

In recent years, human tick-borne disease occurrence has risen in Oklahoma, U.S.A., but year-round data on tick presence in frequently used recreational areas is not widely available. In this study, ticks were collected monthly for one year at residential and non-residential sites in a suburban area of Oklahoma County, OK, U.S.A. At each trapping site, dry ice traps were used in both woodland and grassland areas and fabric tick drags were used in grassland areas. Four species were collected from each park: Amblyomma americanum, Amblyomma maculatum, Dermacentor variabilis, and Ixodes scapularis. Prior to this study, A. americanum was the only species with an established population in Oklahoma County. Consistent with this, A. americanum was collected in all months of the year and accounted for over 90% of ticks collected at each site. Based on our tick survey, we report that A. maculatum, D. variabilis, and I. scapularis, which were each collected in numbers greater than six within a single sampling occasion, are now each confirmed as established populations in Oklahoma County.

Trypanosoma cruzi is a vector-borne protozoan parasite that infects seven million individuals in Central and South America and is the etiologic agent of Chagas disease. There are increasing reports of endemic transmission within the southern US. Trypanosoma cruzi occurs in wild raccoons and dogs in Oklahoma, but its endemicity in the state is poorly studied. We suspected Mexican free-tailed bats ( Tadarida brasiliensis) contributed to the endemicity of T. cruzi in Oklahoma due to their annual migration from Central America to their North American maternity roosts. During the summer of 2017, we sampled 361 Mexican free-tailed bats for T. cruzi at three maternity roosts in Oklahoma. We collected wing tissues, extracted T. cruzi DNA, amplified target DNA by PCR using the primers TCZ1/TCZ2, and observed amplification by gel electrophoresis. One juvenile Mexican free-tailed bat was positive for T. cruzi resulting in a 0.27% prevalence in the 361 sampled bats. Our finding of a wild bat naturally infected with T. cruzi in Oklahoma provided insight on the endemicity of T. cruzi in underrepresented endemic areas. The positive sample was sequenced, confirmed as T. cruzi, and uploaded to GenBank (no. MG869732). Future research will focus on monitoring T. cruzi prevalence in wild bats and insect vectors to better understand the enzootic emergence of this neglected tropical parasite.

This is the first investigation to determine if bats from western Oklahoma have antibodies against Pseudogymnoascus destructans (Pd), the causative agent of White-nose syndrome in bats. The bats examined came from caves near the western most locality in the United States where a bat suspected of having Pd had been reported. A total of fifty-one serum samples were obtained from cave myotis (Myotis velifer), big brown (Eptesicus fuscus) and Townsends big-eared (Corynorhinus townsendii) bats from western Oklahoma between January and February of 2011 and 2012. Nobuto strips containing blood samples from South African bats (Rhinolophus denti, Neoromicia capensis, Hipposideros commersoni, Nycteris thebaica) were used as negative controls. None of the African bats examined appeared to have antibodies against Pd. Thirteen M. velifer (41% of the Oklahoman bats) tested appeared to have antibodies against some strains of Pseudogymnoascus but it is unclear if they have antibodies specifically against Pd.

Misconceptions, or alternative conceptions, are incorrect understandings that students have incorporated into their prior knowledge. The goal of this study was the identification of misconceptions in microbiology held by undergraduate students upon entry into an introductory, general microbiology course. This work was the first step in developing a microbiology concept inventory based on the American Society for Microbiology’s Recommended Curriculum Guidelines for Undergraduate Microbiology. Responses to true/false (T/F) questions accompanied by written explanations by undergraduate students at a diverse set of institutions were used to reveal misconceptions for fundamental microbiology concepts. These data were analyzed to identify the most difficult core concepts, misalignment between explanations and answer choices, and the most common misconceptions for each core concept. From across the core concepts, nineteen misconception themes found in at least 5% of the coded answers for a given question were identified. The top five misconceptions, with coded responses ranging from 19% to 43% of the explanations, are described, along with suggested classroom interventions. Identification of student misconceptions in microbiology provides a foundation upon which to understand students’ prior knowledge and to design appropriate tools for improving instruction in microbiology.

If we are to teach effectively, tools are needed to measure student learning. A widely used method for quickly measuring student understanding of core concepts in a discipline is the concept inventory (CI). Using the American Society for Microbiology Curriculum Guidelines (ASMCG) for microbiology, faculty from 11 academic institutions created and validated a new microbiology concept inventory (MCI). The MCI was developed in three phases. In phase one, learning outcomes and fundamental statements from the ASMCG were used to create T/F questions coupled with open responses. In phase two, the 743 responses to MCI 1.0 were examined to find the most common misconceptions, which were used to create distractors for multiple-choice questions. MCI 2.0 was then administered to 1,043 students. The responses of these students were used to create MCI 3.0, a 23-question CI that measures students’ understanding of all 27 fundamental statements. MCI 3.0 was found to be reliable, with a Cronbach’s alpha score of 0.705 and Ferguson’s delta of 0.97. Test item analysis demonstrated good validity and discriminatory power as judged by item difficulty, item discrimination, and point-biserial correlation coefficient. Comparison of pre- and posttest scores showed that microbiology students at 10 institutions showed an increase in understanding of concepts after instruction, except for questions probing metabolism (average normalized learning gain was 0.15). The MCI will enable quantitative analysis of student learning gains in understanding microbiology, help to identify misconceptions, and point toward areas where efforts should be made to develop teaching approaches to overcome them.

Host control ofCoxiella burnetiiinfections is believed to be mediated primarily by activated monocytes/macrophages. The activation of macrophages by cytokines leads to the production of reactive oxygen intermediates (ROI) and reactive nitrogen intermediates (RNI) that have potent antimicrobial activities. The contributions of ROI and RNI to the inhibition ofC. burnetiireplication were examined in vitro by the use of murine macrophage-like cell lines and primary mouse macrophages. A gamma interferon (IFN-γ) treatment of infected cell lines and primary macrophages resulted in an increased production of nitric oxide (NO) and hydrogen peroxide (H2O2) and a significant inhibition ofC. burnetiireplication. The inhibition of replication was reversed in the murine cell line J774.16 upon the addition of either the inducible nitric oxide synthase (iNOS) inhibitor NG-monomethyl-l-arginine (NGMMLA) or the H2O2scavenger catalase. IFN-γ-treated primary macrophages from iNOS−/−and p47phox−/−mice significantly inhibited replication but were less efficient at controlling infection than IFN-γ-treated wild-type macrophages. To investigate the contributions of ROI and RNI to resistance to infection, we performed in vivo studies, using C57BL/6 wild-type mice and knockout mice lacking iNOS or p47phox. Both iNOS−/−and p47phox−/−mice were attenuated in the ability to controlC. burnetiiinfection compared to wild-type mice. Together, these results strongly support a role for both RNI and ROI in the host control ofC. burnetiiinfection.

Background Chronic wounds such as diabetic foot ulcers, pressure ulcers, and venous leg ulcers contribute to a considerable amount of mortality in the U.S. annually. The inability of these wounds to heal has now been associated with the presence of microbial biofilms. The aim of this study was to determine if products secreted by S. aureus biofilms play an active role in chronic wounds by promoting inflammation, which is a hallmark of chronic wounds. Methods In vitro experiments were conducted to examine changes in gene expression profiles and inflammatory response of human epithelial keratinocytes (HEKa) exposed to products secreted by S. aureus grown in biofilms or products secreted by S. aureus grown planktonically. Results After only two hours of exposure, gene expression microarray data showed marked differences in inflammatory, apoptotic, and nitric oxide responses between HEKa cells exposed to S. aureus biofilm conditioned media (BCM) and HEKa cells exposed to S. aureus planktonic conditioned media (PCM). As early as 4 hours post exposure, ELISA results showed significant increases in IL-6, IL-8, TNFα, and CXCL2 production by HEKa cells exposed to BCM compared to HEKa cells exposed to PCM or controls. Nitric oxide assay data also showed significant increases in nitric oxide production by HEKa cells treated with BCM compared to HEKa cells treated with PCM, or controls. Conclusions Taken together, these results support and extend previous findings that indicate products secreted by S. aureus biofilms directly contribute to the chronic inflammation associated with chronic wounds.

Ground beef patties were packaged in air with: nylon/polyethylene, Saran/polyester/polyethylene, or Saran film overwrap plus a styrofoam tray. Samples were irradiated at 2 kGy by either gamma rays or electron beam, and evaluated for 7 flavor, 3 mouthfeel, and 7 taste attributes by a trained sensory panel. The only difference observed between irradiated and unirradiated samples was that the latter had a more pronounced beef/brothy flavor than irradiated patties. No differences were detected according to packaging material used. Comparing the two sources of irradiation, patties irradiated by gamma rays had more intense cardboardy and soured flavors, and salty and sour tastes than patties irradiated by electron beam.

Ground pork patties were inoculated separately with 10(9) CFU/g each of three strains of Listeria monocytogenes obtained from the National Animal Disease Center (NADC). Inoculated patties were packaged under vacuum and treated at 414 megapascals (60,000 lb/in2) for up to 60 min by high hydrostatic pressure (HHP). Survivors were determined by surface plating onto modified Oxford agar and trypticase soy agar with yeast extract, as well as by the most probable number method using Listeria enrichment broth. Average D values ranged from 1.89 to 4.17 min, depending on the strain, with the most virulent strain (reported by the NADC) having the highest D value. We tested the usefulness of applying a mild heat treatment at 50 degrees C, simultaneously with HHP, to lower these values. Average D values ranged from 0.37 to 0.63 min, depending on the strain. Thus, a 10-log10 reduction could be achieved even in the most pressure-resistant strain of L. monocytogenes by a 6-min application of heat and HHP. Shelf life studies were also conducted, with spoilage levels reached after 5 days of storage at 4 degrees C for controls versus 28 days for treated samples. Sensory evaluation of uninoculated grilled patties showed that panelists could not distinguish between those treated by heat and HHP and untreated controls (P0.05). Thus, treatment by HHP in combination with mild heating can be used successfully to produce safer, longer-lasting fresh pork without affecting quality.

Coxiella burnetii is an obligate intracellular bacterium. The inability to cultivate this organism on axenic medium has made calculation of infectious units challenging and prevents the use of conventional antibiotic susceptibility assays. A rapid and reliable real-time PCR assay was developed to quantify C. burnetii cells from J774.16 mouse macrophage cells and was applied to antibiotic susceptibility testing of C. burnetii Nine Mile, phase I. For calculation of bacterial replication, real-time PCR performed equally as well as immunofluorescent-antibody (IFA) assay when J774.16 cells were infected with 10-fold serial dilutions of C. burnetii and was significantly ( P < 0.05) more repeatable than IFA when 2-fold dilutions were used. Newly infected murine macrophage-like J774.16 cells were treated with 8 μg of chloramphenicol per ml, 4 μg of tetracycline per ml, 4 μg of rifampin per ml, 4 μg of ampicillin per ml, or 1 μg of ciprofloxacin per ml. After 6 days of treatment, tetracycline, rifampin, and ampicillin significantly ( P < 0.01) inhibited the replication of C. burnetii , while chloramphenicol and ciprofloxacin did not. In general, these results are consistent with those from prior reports on the efficacy of these antibiotics against C. burnetii Nine Mile, phase I, and indicate that a real-time PCR-based assay is an appropriate alternative to the present methodology for evaluation of the antibiotic susceptibilities of C. burnetii .

Irradiation of ground beef patties inoculated with the organism Escherichia coli O157:H7 was performed either by gamma rays from a cobalt 60 source or by electron beam generated by a linear accelerator. Patties were packaged in one of the following materials: nylon/polyethylene bags, Saran/polyester/polyethylene bags (PM2), or Saran overwrap with a Styrofoam tray inside. Bags were sealed in air or under vacuum and were irradiated at either 5 or -15 degrees C. Average D10 values (dose required to inactivate 90% of a microbial population) ranged from 0.27 to 0.63 kGy, depending on the conditions. Overall, higher D10 values (P0.0001) were obtained upon irradiation at -15 degrees C as compared with 5 degrees C. Cells inoculated in samples packaged in PM2 had the highest D10 values, but only if irradiated by electron beam at -15 degrees C (P0.001). Since PM2 had the lowest oxygen permeability rate and since the temperature was too low for radicals to migrate easily, these conditions may have minimized the effect of oxygen- and water-derived radicals on microbial survival. Irradiation by gamma rays resulted in higher D10 values (P0.047) than irradiation by electron beam, with the highest values being observed at -15 degrees C. Differences may be attributed to dose rate (1.0 kGy/h for gamma, 17 kGy/min for electron beam) since it is possible that, at low dose rates, microbial enzymes may have more time to repair damage to the cell due to irradiation, resulting in higher D10 values.