40 CFR 503.25 - Operational standards-pathogens and vector attraction reduction.

Code of Federal Regulations, 2010 CFR

2010-07-01

... vector attraction reduction. 503.25 Section 503.25 Protection of Environment ENVIRONMENTAL PROTECTION... § 503.25 Operational standards—pathogens and vector attraction reduction. (a) Pathogens—sewage sludge... active sewage sludge unit, unless the vector attraction reduction requirement in § 503.33(b)(11) is met...

40 CFR 503.25 - Operational standards-pathogens and vector attraction reduction.

Code of Federal Regulations, 2011 CFR

2011-07-01

... vector attraction reduction. 503.25 Section 503.25 Protection of Environment ENVIRONMENTAL PROTECTION... § 503.25 Operational standards—pathogens and vector attraction reduction. (a) Pathogens—sewage sludge... active sewage sludge unit, unless the vector attraction reduction requirement in § 503.33(b)(11) is met...

Development of blood transfusion product pathogen reduction treatments: a review of methods, current applications and demands.

PubMed

Salunkhe, Vishal; van der Meer, Pieter F; de Korte, Dirk; Seghatchian, Jerard; Gutiérrez, Laura

2015-02-01

Transfusion-transmitted infections (TTI) have been greatly reduced in numbers due to the strict donor selection and screening procedures, i.e. the availability of technologies to test donors for endemic infections, and routine vigilance of regulatory authorities in every step of the blood supply chain (collection, processing and storage). However, safety improvement is still a matter of concern because infection zero-risk in transfusion medicine is non-existent. Alternatives are required to assure the safety of the transfusion product and to provide a substitution to systematic blood screening tests, especially in less-developed countries or at the war-field. Furthermore, the increasing mobility of the population due to traveling poses a new challenge in the endemic screening tests routinely used, because non-endemic pathogens might emerge in a specific population. Pathogen reduction treatments sum a plethora of active approaches to eliminate or reduce potential threatening pathogen load from blood transfusion products. Despite the success of pathogen reduction treatments applied to plasma products, there is still a long way to develop and deploy pathogen reduction treatments to cellular transfusion products (such as platelets, RBCs or even to whole blood) and there is divergence on its acceptance worldwide. While the use of pathogen reduction treatments in platelets is performed routinely in a fair number of European blood banks, most of these treatments are not (or just) licensed in the USA or elsewhere in the world. The development of pathogen reduction treatments for RBC and whole blood is still in its infancy and under clinical trials. In this review, we discuss the available and emerging pathogen reduction treatments and their advantages and disadvantages. Furthermore, we highlight the importance of characterizing standard transfusion products with current and emerging approaches (OMICS) and clinical outcome, and integrating this information on a database, thinking on the benefits it might bring in the future toward personalized transfusion therapies. Copyright © 2014 Elsevier Ltd. All rights reserved.

Inactivation of foodborne pathogens in ground beef by cooking with highly controlled radio frequency energy.

PubMed

Schlisselberg, Dov B; Kler, Edna; Kalily, Emmanuel; Kisluk, Guy; Karniel, Ohad; Yaron, Sima

2013-01-01

The consumer demand for fresh tasting, high quality, low salt, preservative-free meals which require minimal preparation time magnifies the safety concern and emphasizes the need to use innovative technologies for food processing. A modern technique to uniformly heat and cook foods is based on a combination of convection and controlled radio frequency (RF) energy. However any advantage conferred on meat cooked by this method would be lost if application of the technology results in decreased safety. Our main goal was to study the inactivation efficacy of this method of cooking against pathogens in ground meat in comparison to standard convection cooking. Meat balls were artificially inoculated with GFP expressing Escherichia coli, Salmonella Typhimurium and Listeria monocytogenes as well as spores of Bacillus cereus and Bacillus thuringiensis and cooked by convection heating (220°C, 40 min), by using energy generated from frequencies in the RF bandwidth (RF cooking, 7.5 min) or by combined heating (5.5 min), until the center temperature of each sample reached 73°C. The mean reductions in total indigenous bacteria obtained by RF and convection were 2.8 and 2.5 log CFU/g, respectively. Cooking of meat balls with convection reduced the E. coli population (8 log CFU/g) by 5.5 log CFU/g, whilst treatment with RF reduced E. coli population to undetectable levels. The mean reductions of S. Typhimurium obtained by RF and convection were 5.7 and 6.5 log CFU/g, respectively. The combined treatment reduced the Salmonella population to undetectable levels. In contrast, L. monocytogenes was poorly affected by RF cooking. The mean reduction of L. monocytogenes obtained by RF energy was 0.4 log CFU/g, while convection cooking resulted in undetectable levels. Interestingly, the combined treatment also resulted with undetectable levels of Listeria although time of cooking was reduced by 86%. One-step cooking had negligible effects on the Bacillus spores and therefore a 2-step treatment of RF or convection was applied. This 2-step treatment proved to be efficient with 4.5 log CFU/g reduction for both RF and convection. In conclusion, here we show that combination of RF with convection cooking resulted in similar or even better effects on selected foodborne pathogens compared to convection only, while the time required for safe cooking is cut down by up to 86%. The equal or better results in the levels of all investigated pathogens using RF with convection compared with convection only suggest that this technology looks promising and safe for ground beef cooking. Copyright © 2012 Elsevier B.V. All rights reserved.

Two pathogen reduction technologies--methylene blue plus light and shortwave ultraviolet light--effectively inactivate hepatitis C virus in blood products.

PubMed

Steinmann, Eike; Gravemann, Ute; Friesland, Martina; Doerrbecker, Juliane; Müller, Thomas H; Pietschmann, Thomas; Seltsam, Axel

2013-05-01

Contamination of blood products with hepatitis C virus (HCV) can cause infections resulting in acute and chronic liver diseases. Pathogen reduction methods such as photodynamic treatment with methylene blue (MB) plus visible light as well as irradiation with shortwave ultraviolet (UVC) light were developed to inactivate viruses and other pathogens in plasma and platelet concentrates (PCs), respectively. So far, their inactivation capacities for HCV have only been tested in inactivation studies using model viruses for HCV. Recently, a HCV infection system for the propagation of infectious HCV in cell culture was developed. Inactivation studies were performed with cell culture-derived HCV and bovine viral diarrhea virus (BVDV), a model for HCV. Plasma units or PCs were spiked with high titers of cell culture-grown viruses. After treatment of the blood units with MB plus light (Theraflex MB-Plasma system, MacoPharma) or UVC (Theraflex UV-Platelets system, MacoPharma), residual viral infectivity was assessed using sensitive cell culture systems. HCV was sensitive to inactivation by both pathogen reduction procedures. HCV in plasma was efficiently inactivated by MB plus light below the detection limit already by 1/12 of the full light dose. HCV in PCs was inactivated by UVC irradiation with a reduction factor of more than 5 log. BVDV was less sensitive to the two pathogen reduction methods. Functional assays with human HCV offer an efficient tool to directly assess the inactivation capacity of pathogen reduction procedures. Pathogen reduction technologies such as MB plus light treatment and UVC irradiation have the potential to significantly reduce transfusion-transmitted HCV infections. © 2012 American Association of Blood Banks.

Pathogen reduction co-benefits of nutrient best management practices

PubMed Central

Wainger, Lisa A.; Barber, Mary C.

2016-01-01

Background Many of the practices currently underway to reduce nitrogen, phosphorus, and sediment loads entering the Chesapeake Bay have also been observed to support reduction of disease-causing pathogen loadings. We quantify how implementation of these practices, proposed to meet the nutrient and sediment caps prescribed by the Total Maximum Daily Load (TMDL), could reduce pathogen loadings and provide public health co-benefits within the Chesapeake Bay system. Methods We used published data on the pathogen reduction potential of management practices and baseline fecal coliform loadings estimated as part of prior modeling to estimate the reduction in pathogen loadings to the mainstem Potomac River and Chesapeake Bay attributable to practices implemented as part of the TMDL. We then compare the estimates with the baseline loadings of fecal coliform loadings to estimate the total pathogen reduction potential of the TMDL. Results We estimate that the TMDL practices have the potential to decrease disease-causing pathogen loads from all point and non-point sources to the mainstem Potomac River and the entire Chesapeake Bay watershed by 19% and 27%, respectively. These numbers are likely to be underestimates due to data limitations that forced us to omit some practices from analysis. Discussion Based on known impairments and disease incidence rates, we conclude that efforts to reduce nutrients may create substantial health co-benefits by improving the safety of water-contact recreation and seafood consumption. PMID:27904807

Pathogen reduction co-benefits of nutrient best management practices.

PubMed

Richkus, Jennifer; Wainger, Lisa A; Barber, Mary C

2016-01-01

Many of the practices currently underway to reduce nitrogen, phosphorus, and sediment loads entering the Chesapeake Bay have also been observed to support reduction of disease-causing pathogen loadings. We quantify how implementation of these practices, proposed to meet the nutrient and sediment caps prescribed by the Total Maximum Daily Load (TMDL), could reduce pathogen loadings and provide public health co-benefits within the Chesapeake Bay system. We used published data on the pathogen reduction potential of management practices and baseline fecal coliform loadings estimated as part of prior modeling to estimate the reduction in pathogen loadings to the mainstem Potomac River and Chesapeake Bay attributable to practices implemented as part of the TMDL. We then compare the estimates with the baseline loadings of fecal coliform loadings to estimate the total pathogen reduction potential of the TMDL. We estimate that the TMDL practices have the potential to decrease disease-causing pathogen loads from all point and non-point sources to the mainstem Potomac River and the entire Chesapeake Bay watershed by 19% and 27%, respectively. These numbers are likely to be underestimates due to data limitations that forced us to omit some practices from analysis. Based on known impairments and disease incidence rates, we conclude that efforts to reduce nutrients may create substantial health co-benefits by improving the safety of water-contact recreation and seafood consumption.

Efficacy of chemical treatments in eliminating Salmonella and Escherichia coli O157:H7 on scarified and polished alfalfa seeds.

PubMed

Holliday, S L; Scouten, A J; Beuchat, L R

2001-10-01

Alfalfa seeds are sometimes subjected to a scarification treatment to enhance water uptake, which results in more rapid and uniform germination during sprout production. It has been hypothesized that this mechanical abrasion treatment diminishes the efficacy of chemical treatments used to kill or remove pathogenic bacteria from seeds. A study was done to compare the effectiveness of chlorine (20,000 ppm), H2O, (8%), Ca(OH)2 (1%), Ca(OH)2 (1%) plus Tween 80 (1%), and Ca(OH)2 (1%) plus Span 20 (1%) treatments in killing Salmonella and Escherichia coli O157:H7 inoculated onto control, scarified, and polished alfalfa seeds obtained from two suppliers. The influence of the presence of organic material in the inoculum carrier on the efficacy of sanitizers was investigated. Overall, treatment with 1% Ca(OH)2 was the most effective in reducing populations of the pathogens. Reduction in populations of pathogens on seeds obtained from supplier I indicate that chemical treatments are less efficacious in eliminating the pathogens on scarified seeds compared to control seeds. However, the effectiveness of chemical treatment in removing Salmonella and E. coli O157:H7 from seeds obtained from supplier 2 was not markedly affected by scarification or polishing. The presence of organic material in the inoculum carrier did not have a marked influence on the efficacy of chemicals in reducing populations of test pathogens. Additional lots of control, scarified, and polished alfalfa seeds of additional varieties need to be tested before conclusions can be drawn concerning the impact of mechanical abrasion on the efficacy of chemical treatment in removing or killing Salmonella and E. coli O157:H7.

76 FR 30176 - Expedited Review for New Animal Drug Applications for Human Pathogen Reduction Claims; Withdrawal...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-05-24

... DEPARTMENT OF HEALTH AND HUMAN SERVICES Food and Drug Administration [Docket No. FDA-2001-D-0066] (Formerly Docket No. 2001D-0107) Expedited Review for New Animal Drug Applications for Human Pathogen... Review for New Animal Drug Applications for Human Pathogen Reduction Claims.'' The guidance predates the...

Control of foodborne pathogens on fresh-cut fruit by a novel strain of Pseudomonas graminis.

PubMed

Alegre, Isabel; Viñas, Inmaculada; Usall, Josep; Teixidó, Neus; Figge, Marian J; Abadias, Maribel

2013-06-01

The consumption of fresh-cut fruit has substantially risen over the last few years, leading to an increase in the number of outbreaks associated with fruit. Moreover, consumers are currently demanding wholesome, fresh-like, safe foods without added chemicals. As a response, the aim of this study was to determine if the naturally occurring microorganisms on fruit are "competitive with" or "antagonistic to" potentially encountered pathogens. Of the 97 and 107 isolates tested by co-inoculation with Escherichia coli O157:H7, Salmonella and Listeria innocua on fresh-cut apple and peach, respectively, and stored at 20 °C, seven showed a strong antagonistic capacity (more than 1-log unit reduction). One of the isolates, CPA-7, achieved the best reduction values (from 2.8 to 5.9-log units) and was the only isolate able to inhibit E. coli O157:H7 at refrigeration temperatures on both fruits. Therefore, CPA-7 was selected for further assays. Dose-response assays showed that CPA-7 should be present in at least the same amount as the pathogen to adequately reduce the numbers of the pathogen. From the results obtained in in vitro assays, competition seemed to be CPA-7's mode of action against E. coli O157:H7. The CPA-7 strain was identified as Pseudomonas graminis. Thus, the results support the potential use of CPA-7 as a bioprotective agent against foodborne pathogens in minimally processed fruit. Copyright © 2013 Elsevier Ltd. All rights reserved.

Riboflavin-ultraviolet light pathogen reduction treatment does not impact the immunogenicity of murine red blood cells.

PubMed

Tormey, Christopher A; Santhanakrishnan, Manjula; Smith, Nicole H; Liu, Jingchun; Marschner, Susanne; Goodrich, Raymond P; Hendrickson, Jeanne E

2016-04-01

Ultraviolet (UV) illumination/pathogen reduction effectively inactivates white blood cells (WBCs) in whole blood. Given that cotransfused WBCs may impact recipient immune responses, we hypothesized that pathogen reduction of whole blood may alter responses to RBC antigens. Transgenic mice expressing a model (HOD) antigen, authentic human (hGPA or KEL) antigens, or natural fluorescence (uGFP) on their RBCs were utilized as blood donors. Recipients were transfused with fresh whole blood to which riboflavin had been added or fresh whole blood treated by UV illumination/pathogen reduction treatment after the addition of riboflavin. Posttransfusion RBC recovery, survival, and alloimmunization were measured by flow cytometry. UV illumination/pathogen reduction treatment did not alter RBC antigen expression, and recipients of treated syngeneic RBCs had persistently negative direct antiglobulin tests. Greater than 75% of treated and untreated syngeneic RBCs were recovered 24 hours posttransfusion in all experiments, although alterations in the long-term posttransfusion survival of treated RBCs were observed. Treated and untreated KEL RBCs induced similar recipient alloimmune responses, with all recipients making anti-KEL glycoprotein immunoglobulins (p > 0.05). Alloimmune responses to treated HOD or hGPA RBCs were no different from untreated RBCs (p > 0.05). Pathogen inactivation treatment of fresh whole murine blood with riboflavin and UV illumination does not impact the rate or magnitude of RBC alloimmunization to three distinct RBC antigens. Further, UV illumination/pathogen reduction appears safe from an immunohematologic standpoint, with no immunogenic neoantigens detected on treated murine RBCs. Future studies with fresh and stored human RBCs are warranted to confirm these findings. © 2015 AABB.

A novel microfluidic mixer-based approach for determining inactivation kinetics of Escherichia coli O157:H7 in chlorine solutions.

PubMed

Zhang, Boce; Luo, Yaguang; Zhou, Bin; Wang, Qin; Millner, Patricia D

2015-08-01

Determination of the minimum free chlorine concentration needed to prevent pathogen survival/cross-contamination during produce washing is essential for the development of science-based food safety regulations and practices. Although the trend of chlorine concentration-contact time on pathogen inactivation is generally understood, specific information on chlorine and the kinetics of pathogen inactivation at less than 1.00 s is urgently needed by the produce processing industry. However, conventional approaches to obtain this critical data have been unable to adequately measure very rapid responses. This paper reports our development, fabrication, and test of a novel microfluidic device, and its application to obtain the necessary data on pathogen inactivation by free chlorine in produce wash solution in times as short as 0.10 s. A novel microfluidic mixer with the capability to accurately determine the reaction time and control the chlorine concentration was designed with three inlets for bacterial, chlorine and dechlorinating solutions, and one outlet for effluent collection. The master mold was fabricated on a silicon wafer with microchannels via photopolymerization. Polydimethylsiloxane replicas with patterned microchannels were prototyped via soft lithography. The replicas were further assembled into the micromixer on glass via O2 plasma treatment, and the inlets were connected to a syringe pump for solution delivery. To determine the kinetics of free chlorine on pathogen inactivation, chlorine solutions of varying concentrations were first pumped into the micromixer, together with the addition of bacterial suspension of Escherichia coli O157:H7 through a separate inlet. This was followed by injection of dechlorinating solution to stop the chlorine-pathogen reaction. The effluent was collected and the surviving bacteria cells were enumerated using a modified 'Most Probable Number' method. Free chlorine concentration was determined using a standard colorimetric method. The contact time was experimentally set by adjusting the solution flow rate, and was estimated by computational fluid dynamics modeling. Results showed that 1) pathogen inactivation was significantly affected by free chlorine concentration (P < 0.0001) and subsecond reaction time (P < 0.0001) and their interactions (P < 0.0001); and 2) the current industry practice of using 1.0 mg/L free chlorine will require more than 1.00 s total contact to achieve a 5-log10 reduction in an E. coli O157:H7 population, whereas a 10.0 mg/L free chlorine solution will achieve 5-log10 reduction in as little as 0.25 s. Information obtained from this study will provide critical insight on kinetics of bacterial inactivation for a broad range of sanitizers and produce wash operational conditions, thus facilitating the development and implementation of science-based food safety regulations and practices for improving food safety. Published by Elsevier Ltd.

Pathogen inactivation techniques.

PubMed

Pelletier, J P R; Transue, S; Snyder, E L

2006-01-01

The desire to rid the blood supply of pathogens of all types has led to the development of many technologies aimed at the same goal--eradication of the pathogen(s) without harming the blood cells or generating toxic chemical agents. This is a very ambitious goal, and one that has yet to be achieved. One approach is to shun the 'one size fits all' concept and to target pathogen-reduction agents at the Individual component types. This permits the development of technologies that might be compatible with, for example, plasma products but that would be cytocidal and thus incompatible with platelet concentrates or red blood cell units. The technologies to be discussed include solvent detergent and methylene blue treatments--designed to inactivate plasma components and derivatives; psoralens (S-59--amotosalen) designed to pathogen-reduce units of platelets; and two products aimed at red blood cells, S-303 (a Frale--frangible anchor-linker effector compound) and Inactine (a binary ethyleneimine). A final pathogen-reduction material that might actually allow one material to inactivate all three blood components--riboflavin (vitamin B2)--is also under development. The sites of action of the amotosalen (S-59), the S-303 Frale, Inactine, and riboflavin are all localized in the nucleic acid part of the pathogen. Solvent detergent materials act by dissolving the plasma envelope, thus compromising the integrity of the pathogen membrane and rendering it non-infectious. By disrupting the pathogen's ability to replicate or survive, its infectivity is removed. The degree to which bacteria and viruses are affected by a particular pathogen-reducing technology relates to its Gram-positive or Gram-negative status, to the sporulation characteristics for bacteria, and the presence of lipid or protein envelopes for viruses. Concerns related to photoproducts and other breakdown products of these technologies remain, and the toxicology of pathogen-reduction treatments is a major ongoing area of investigation. Clearly, regulatory agencies have a major role to play in the evaluation of these new technologies. This chapter will cover the several types of pathogen-reduction systems, mechanisms of action, the inactivation efficacy for specific types of pathogens, toxicology of the various systems and the published research and clinical trial data supporting their potential usefulness. Due to the nature of the field, pathogen reduction is a work in progress and this review should be considered as a snapshot in time rather than a clear picture of what the future will bring.

75 FR 30844 - General Mills, Inc.; Withdrawal of Food Additive Petition

Federal Register 2010, 2011, 2012, 2013, 2014

2010-06-02

... for the reduction of pathogens and other microorganisms in aqueous sugar solutions and potable water... reduction of pathogens and other microorganisms in aqueous sugar solutions and potable water intended for...

Inactivation of selected bacterial pathogens in dairy cattle manure by mesophilic anaerobic digestion (balloon type digester).

PubMed

Manyi-Loh, Christy E; Mamphweli, Sampson N; Meyer, Edson L; Okoh, Anthony I; Makaka, Golden; Simon, Michael

2014-07-14

Anaerobic digestion of animal manure in biogas digesters has shown promise as a technology in reducing the microbial load to safe and recommended levels. We sought to treat dairy manure obtained from the Fort Hare Dairy Farm by investigating the survival rates of bacterial pathogens, through a total viable plate count method, before, during and after mesophilic anaerobic digestion. Different microbiological media were inoculated with different serial dilutions of manure samples that were withdrawn from the biogas digester at 3, 7 and 14 day intervals to determine the viable cells. Data obtained indicated that the pathogens of public health importance were 90%-99% reduced in the order: Campylobacter sp. (18 days) < Escherichia coli sp. (62 days) < Salmonella sp. (133 days) from a viable count of 10.1 × 103, 3.6 × 105, 7.4 × 103 to concentrations below the detection limit (DL = 102 cfu/g manure), respectively. This disparity in survival rates may be influenced by the inherent characteristics of these bacteria, available nutrients as well as the stages of the anaerobic digestion process. In addition, the highest p-value i.e., 0.957 for E. coli showed the statistical significance of its model and the strongest correlation between its reductions with days of digestion. In conclusion, the results demonstrated that the specific bacterial pathogens in manure can be considerably reduced through anaerobic digestion after 133 days.

Proving the antimicrobial spectrum of an amphoteric surfactant-sol-gel coating: a food-borne pathogen study.

PubMed

Copello, G J; Teves, S; Degrossi, J; D'Aquino, M; Desimone, M F; Díaz, L E

2008-09-01

An antimicrobial coating was evaluated in this work for its antimicrobial efficacy against common food-borne pathogens. Dodecyl-di(aminoethyl)-glycine, an organic disinfectant, was immobilized in a silicon oxide matrix to generate thin films over surfaces by means of the sol-gel process. Tetraethoxysilane was used as the polymeric precursor. No alteration of optical transparency on the covered surfaces was observed. Topographic images obtained with atomic force microscopy showed a homogeneous film with no additional roughness added by the polymer to the surface. The attenuated total reflectance-Fourier transform infrared spectral data showed the presence of dodecyl-di(aminoethyl)-glycine in the silicon oxide network after a normal cleaning procedure. The antimicrobial efficacy test was performed by exposing coated slides to suspensions of common food-borne pathogens: Escherichia coli, Staphyloccocus aureus, E. coli O157:H7, Salmonella typhi, S. cholerasuiss, Listeria innocua and L. monocytogenes. The coating activity was not only bacteriostatic but also bactericidal. The percent reduction of viable microorganism exposure over 24 h to the coated surface ranged between 99.5%, for the more resistant gram-positive bacteria, and over 99.999%, for most gram-negative bacteria. The silicon matrix itself did not account for any reduction of viable microbial, even more an increase was observed.

Opportunities for mitigating pathogen contamination during on-farm food production.

PubMed

Doyle, Michael P; Erickson, Marilyn C

2012-01-16

Fruits, vegetables, and meat are susceptible to contamination by foodborne pathogens at many points from production through preparation in the home. This review will largely highlight approaches and progress made in the last five years to address strategies to reduce pathogen contamination in animal production but will also touch on the emerging field of preharvest produce food safety. Mitigation strategies can be divided into those that address pathogen reduction in the environment and those that target reduction/elimination of pathogen contamination in animals or plants. The former strategy has been encompassed in studies evaluating sanitation treatments of facilities as well as in numerous epidemiologic risk assessment studies (both on-farm assessments and computer simulation models) that identify management practices that impact pathogen prevalence in animals. Interventions to significantly reduce pathogen exposure via feed or water are dependent on their role as a significant contributor to pathogen contamination in the animal production system. In addition, inconsistent results obtained with interventions of dietary additives or formulation modifications (grain versus forage; inclusion of distiller's grains) on pathogen prevalence in animals have been attributed to a range of factors including target organism, grain type, level of inclusion, the animal's health or stress level, and ability to survive the gastric acidic conditions. Recent attempts to microencapsulate organic acids or bacteriophage within feed have met with only marginal improvements in reducing pathogen carriage in animals but this approach may have greater potential with other antimicrobial additives (i.e., essential oils). Bacteriophage therapy, in general, can significantly reduce pathogen carriage in animals but based on its transient nature and the potential for development of phage-resistant subpopulations, this approach should be administered to animals just prior to slaughter and preferably to animals that are suspected "super-shedders". Other promising on-farm intervention approaches have included breeding for pathogen resistance, vaccines, and dietary bacteriocins. To optimize interventions on a cost basis, studies have also determined that application of dietary interventions at specific time points in the animal's production cycle is a useful strategy to reduce pathogen carriage (e.g., probiotics to fertilized eggs and acidified feed to fattening swine). In conclusion, applicable management and intervention strategies may vary depending on the type of food under production; however, it is important to consider from a holistic view how any new intervention strategies will affect the overall production system in order to maintain a successful, efficient food production environment. Copyright © 2011 Elsevier B.V. All rights reserved.

40 CFR 503.15 - Operational standards-pathogens and vector attraction reduction.

Code of Federal Regulations, 2014 CFR

2014-07-01

... met when bulk sewage sludge is applied to a lawn or a home garden. (3) The Class A pathogen... home garden. (3) One of the vector attraction reduction requirements in § 503.33 (b)(1) through (b)(8...

40 CFR 503.15 - Operational standards-pathogens and vector attraction reduction.

Code of Federal Regulations, 2013 CFR

2013-07-01

... met when bulk sewage sludge is applied to a lawn or a home garden. (3) The Class A pathogen... home garden. (3) One of the vector attraction reduction requirements in § 503.33 (b)(1) through (b)(8...

40 CFR 503.15 - Operational standards-pathogens and vector attraction reduction.

Code of Federal Regulations, 2012 CFR

2012-07-01

... met when bulk sewage sludge is applied to a lawn or a home garden. (3) The Class A pathogen... home garden. (3) One of the vector attraction reduction requirements in § 503.33 (b)(1) through (b)(8...

Inactivation of viruses and bacteria on strawberries using a levulinic acid plus sodium dodecyl sulfate based sanitizer, taking sensorial and chemical food safety aspects into account.

PubMed

Zhou, Zijin; Zuber, Sophie; Cantergiani, Frédérique; Butot, Sophie; Li, Dan; Stroheker, Thomas; Devlieghere, Frank; Lima, Anthony; Piantini, Umberto; Uyttendaele, Mieke

2017-09-18

The efficacy of levulinic acid (LVA) in combination with sodium dodecyl sulfate (SDS) in removal of foodborne viruses, enteric bacterial pathogens and their surrogates on fresh strawberries was investigated. Inoculated strawberries were treated with potable water, sodium hypochlorite solution (50ppm), 0.5% LVA plus 0.5% SDS solution, and 5% LVA plus 2% SDS solution respectively for 2min, followed by spray-rinsing with potable water. Water washing removed at least 1.0-log of the tested viral and bacterial strains from the strawberries' surfaces. The 50ppm chlorine wash induced 3.4, 1.5 and 2.1-log reductions for hepatitis A virus (HAV), murine norovirus-1 (MNV-1) and MS2 bacteriophage, respectively. In comparison, the tested bacterial strains showed uniform reductions around 1.6-log CFU/ml. The 0.5% LVA plus 0.5% SDS wash induced 2.7, 1.4 and 2.4-log reductions for HAV, MNV-1 and MS2, which were comparable with the reductions induced by chlorine (P>0.05). For bacteria, over 2.0-log reductions were obtained for Enterococcus faecium, Listeria monocytogenes and Salmonella, while Escherichia coli O157:H7 and Escherichia coli P1 showed reductions of 1.9 and 1.8-log CFU/ml. Higher concentration of LVA plus SDS showed no significantly higher reductions (P>0.05). Sensory tests of washed strawberries and chemical residue analysis of LVA on strawberries after washing were also performed. In conclusion, this study demonstrates good performance of 0.5% LVA plus 0.5% SDS to reduce the levels of enteric pathogens if present on strawberries without altering taste and introducing chemical safety issues. Copyright © 2017 Elsevier B.V. All rights reserved.

Risk-based enteric pathogen reduction targets for non-potable ...

EPA Pesticide Factsheets

This paper presents risk-based enteric pathogen log reduction targets for non-potable and potable uses of a variety of alternative source waters (i.e., locally-collected greywater, roof runoff, and stormwater). A probabilistic Quantitative Microbial Risk Assessment (QMRA) was used to derive the pathogen log10 reduction targets (LRTs) that corresponded with an infection risk of either 10−4 per person per year (ppy) or 10−2 ppy. The QMRA accounted for variation in pathogen concentration and sporadic pathogen occurrence (when data were available) in source waters for reference pathogens in the genera Rotavirus, Mastadenovirus (human adenoviruses), Norovirus, Campylobacter, Salmonella, Giardia and Cryptosporidium. Non-potable uses included indoor use (for toilet flushing and clothes washing) with occasional accidental ingestion of treated non-potable water (or cross-connection with potable water), and unrestricted irrigation for outdoor use. Various exposure scenarios captured the uncertainty from key inputs, i.e., the pathogen concentration in source water; the volume of water ingested; and for the indoor use, the frequency of and the fraction of the population exposed to accidental ingestion. Both potable and non-potable uses required pathogen treatment for the selected waters and the LRT was generally greater for potable use than non-potable indoor use and unrestricted irrigation. The difference in treatment requirements among source waters was driven by the

Use of the Blom Tracheotomy Tube with Suction Inner Cannula to Decontaminate Microorganisms from the Subglottic Space. A Proof of Concept.

PubMed

Rabach, Lesley; Siegel, Mark D; Puchalski, Jonathan T; Towle, Dana; Follert, Michelle; Johnson, Kelsey M; Rademaker, Alfred W; Leder, Steven B

2015-06-01

Preventing pulmonary complications during mechanical ventilation via tracheotomy is a high priority. To investigate if the Blom tracheotomy tube with suction-above-the-cuff inner cannula reduced the quantity of normal flora and pathogens in supra- versus subglottic spaces. We enrolled 20 consecutive medical ICU adults requiring tracheostomy for mechanical ventilation in this proof-of-concept, prospective, single-center study. All participants received a Blom tracheotomy tube with suction-above-the-cuff inner cannula to decontaminate microorganisms from the supra- and subglottic spaces. Supra- and subglottic sputum samples were obtained for microbiologic analysis while an endotracheal tube was in place before tracheotomy and once per week for up to 4 weeks of mechanical ventilation after tracheotomy. Demographics, duration of endotracheal tube intubation, and duration of mechanical ventilation post-tracheotomy were recorded. There was a significant reduction for supraglottic (2.86 ± 1.11 [mean ± SD]) versus subglottic suction samples (2.48 ± 1.07) (paired t test, P = 0.048; Wilcoxon test, P = 0.045) when all data pairs for normal flora and pathogens were combined across times. There was a significant reduction of normal flora pooled across times in 19 data pairs for supraglottic (3.00 ± 1.05) versus subglottic suction samples (2.00 ± 0.94) (paired t test, P = 0.0004; Wilcoxon test, P = 0.0007). There was no significant reduction of pathogens pooled across times in 25 data pairs for supraglottic (2.76 ± 1.16) versus subglottic suction samples (2.84 ± 1.03) (paired t test, P = 0.75; Wilcoxon test, P = 0.83). Proof-of-concept was confirmed. The Blom tracheotomy tube with disposable suction-above-the-cuff inner cannula decontaminated microorganisms from the subglottic space when normal flora and pathogens were combined. Future research should investigate if decreased quantity of normal flora and pathogens in the subglottic space reduces the incidence of ventilator-associated pulmonary complications in critically ill patients requiring ongoing mechanical ventilation via tracheotomy.

Removal of pathogenic bacteria from sewage-treated effluent and biosolids for agricultural purposes

NASA Astrophysics Data System (ADS)

Al-Gheethi, A. A.; Efaq, A. N.; Bala, J. D.; Norli, I.; Abdel-Monem, M. O.; Ab. Kadir, M. O.

2018-05-01

The reuse of treated sewage for irrigation is considered as an important alternative water source in the new water management strategy of the countries that face a severe deficiency of water resources such as the Middle East countries. The organic material and fertilizing elements contained in biosolids are essential for maintaining soil fertility. However, both treated sewage and biosolids contain a large diversity of pathogens that would be transmitted to the environment and infect human directly or indirectly. Therefore, those pathogens should be reduced from the treated sewage and biosolids before the reuse in the agriculture. This paper reviews the considerations for reuse of treated sewage and biosolids in agriculture and further treatments used for reduction of pathogenic bacteria. The treatment methods used for the reduction of pathogens in these wastes have reviewed. It appeared that the main concern associated with the reduction of pathogenic bacteria lies in their ability to regrow in the treated sewage and biosolids. Therefore, the effective treatment method is that it has the potential to destruct pathogens cells and remove the nutrients to prevent the regrowth or recontamination from the surrounded environment. The removal of nutrients might be applicable in the sewage but not in the biosolids due to high nutrient contents. However, the reduction of health risk in the biosolids might be carried out by regulating the biosolid utilization and selecting the plant species grown in the fertilized soil with biosolids.

Escherichia albertii Inactivation following l-Lactic Acid Exposure or Cooking in Ground Beef.

PubMed

Jones-Ibarra, Amie M; Wall, Kayley R; Vuia-Riser, Jennifer; Kerth, Chris R; Castillo, Alejandro; Taylor, T Matthew

2016-09-01

Escherichia albertii is an emerging foodborne pathogen recovered from young children and adults exhibiting symptoms of gastroenteritis via pathogenesis factors including attaching and effacing lesions, cytolethal distending toxin, and Shiga toxin variants. Study objectives were to determine E. albertii survival following (i) exposure to lactic acid as a function of solution pH and incubation period and (ii) cooking ground beef patties to different endpoint temperatures. E. albertii was incubated in phosphate buffer containing 3.0% l-lactic acid adjusted to pH 3.0, 4.0, 5.0, or 7.0; survivors were determined every 30 min for 150 min. Ground beef patties (80% lean) were cooked to temperature endpoints simulating undercooking (62°C), the minimum temperature for safe cooking (71.1°C), and cooking to well done (76°C). Maximal pathogen reduction was observed after a 30-min exposure to pH 3.0 l-lactic acid. Reductions of 3.9, 4.4, and 4.9 log CFU/g were obtained following cooking ground beef patties to 62, 71.1, and 76°C, respectively, but the reductions did not differ as a function of the endpoint cooking temperature (P ≥ 0.05). E. albertii may be controlled on beef through the proper application of antimicrobial interventions and cooking.

Pathogen reduction in minimally managed composting of bovine manure.

PubMed

Millner, Patricia; Ingram, David; Mulbry, Walter; Arikan, Osman A

2014-11-01

Spread of manure pathogens is of considerable concern due to use of manure for land application. In this study, the effects of four static pile treatment options for bovine manure on die-off of a generic Escherichia coli, E. coli O157:H7 surrogate, Salmonella Senftenberg, Salm. Typhimurium, and Listeria monocytogenes were evaluated. Bovine manure spiked with these bacteria were placed in cassettes at the top, middle, and bottom sections of four static pile treatments that reflect minimal changes in pile construction with and without straw. Temperatures were monitored continuously during the 28 day self-heating period. E. coli and salmonellae were reduced from 8 to 9 log10 CFU g(-1) to undetectable levels (<1.77 log10 MPN g(-1)) at 25-30 cm depths within 7 days in all pile sections except for the manure-only pile in which 3-4 logs of reduction were obtained. No L. monocytogenes initially present at 6.62 log10 CFU g(-1) were recovered from straw-amended piles after 14 days, in contrast with manure-only treatment in which this pathogen was recovered even at 28 days. Decline of target bacterial populations corresponded to exposure to temperatures above 45°C for more than 3 days and amendments of manure with straw to increase thermophilic zones. Use of straw to increase aeration, self-heating capacity, and heat retention in manure piles provides producers a minimal management option for composting that enhances pathogen die-off and thereby reduces risk of environmental spread when manure is applied to land. Published by Elsevier Ltd.

Significance of the Bacteriophage Treatment Schedule in Reducing Salmonella Colonization of Poultry

PubMed Central

Bardina, Carlota; Spricigo, Denis A.; Cortés, Pilar

2012-01-01

Salmonella remains the major cause of food-borne diseases worldwide, with chickens known to be the main reservoir for this zoonotic pathogen. Among the many approaches to reducing Salmonella colonization of broilers, bacteriophage offers several advantages. In this study, three bacteriophages (UAB_Phi20, UAB_Phi78, and UAB_Phi87) obtained from our collection that exhibited a broad host range against Salmonella enterica serovar Enteritidis and Salmonella enterica serovar Typhimurium were characterized with respect to morphology, genome size, and restriction patterns. A cocktail composed of the three bacteriophages was more effective in promoting the lysis of S. Enteritidis and S. Typhimurium cultures than any of the three bacteriophages alone. In addition, the cocktail was able to lyse the Salmonella enterica serovars Virchow, Hadar, and Infantis. The effectiveness of the bacteriophage cocktail in reducing the concentration of S. Typhimurium was tested in two animal models using different treatment schedules. In the mouse model, 50% survival was obtained when the cocktail was administered simultaneously with bacterial infection and again at 6, 24, and 30 h postinfection. Likewise, in the White Leghorn chicken specific-pathogen-free (SPF) model, the best results, defined as a reduction of Salmonella concentration in the chicken cecum, were obtained when the bacteriophage cocktail was administered 1 day before or just after bacterial infection and then again on different days postinfection. Our results show that frequent treatment of the chickens with bacteriophage, and especially prior to colonization of the intestinal tract by Salmonella, is required to achieve effective bacterial reduction over time. PMID:22773654

Inactivation of Selected Bacterial Pathogens in Dairy Cattle Manure by Mesophilic Anaerobic Digestion (Balloon Type Digester)

PubMed Central

Manyi-Loh, Christy E.; Mamphweli, Sampson N.; Meyer, Edson L.; Okoh, Anthony I.; Makaka, Golden; Simon, Michael

2014-01-01

Anaerobic digestion of animal manure in biogas digesters has shown promise as a technology in reducing the microbial load to safe and recommended levels. We sought to treat dairy manure obtained from the Fort Hare Dairy Farm by investigating the survival rates of bacterial pathogens, through a total viable plate count method, before, during and after mesophilic anaerobic digestion. Different microbiological media were inoculated with different serial dilutions of manure samples that were withdrawn from the biogas digester at 3, 7 and 14 day intervals to determine the viable cells. Data obtained indicated that the pathogens of public health importance were 90%–99% reduced in the order: Campylobacter sp. (18 days) < Escherichia coli sp. (62 days) < Salmonella sp. (133 days) from a viable count of 10.1 × 103, 3.6 × 105, 7.4 × 103 to concentrations below the detection limit (DL = 102 cfu/g manure), respectively. This disparity in survival rates may be influenced by the inherent characteristics of these bacteria, available nutrients as well as the stages of the anaerobic digestion process. In addition, the highest p-value i.e., 0.957 for E. coli showed the statistical significance of its model and the strongest correlation between its reductions with days of digestion. In conclusion, the results demonstrated that the specific bacterial pathogens in manure can be considerably reduced through anaerobic digestion after 133 days. PMID:25026086

Selection of potential biological control of Exserohilum turcicum with epiphytic microorganisms from maize.

PubMed

Sartori, Melina; Nesci, Andrea; Formento, Ángela; Etcheverry, Miriam

2015-01-01

The aims of this study were to select microbial isolates from phyllosphere of maize and to examine their antagonistic activity against Exserohilum turcicum. Selection was performed through the ability of isolates to compete with the pathogen using an index of dominance and to affect growth parameters of E. turcicum. Most of the epiphytic populations obtained for the screening were bacteria. These isolates were found in the order of 6 log CFU/g of leaf fresh weight. According to similar morphological characteristics and staining, 44 out of 111 isolates obtained were selected for testing antagonistic effects. At water potential, ψ, -1.38MPa and -4.19MPa, three Bacillus isolates showed dominance at a distance (5/0) and a significant reduction of growth rate of the pathogen. Three Bacillus isolates only decreased the growth rate of E. turcicum at -1.38MPa. At -4.19MPa the growth rate decreased with three isolates of Pantoea and three Bacillus. In this study a negative and significant correlation was observed between the growth rate of E. turcicum and the dominance index in the interaction of the pathogen with some bacteria. These results show that with decreasing growth rate of the pathogen the dominance index of the interaction increases. Eleven potential biocontrol agents against E. turcicum were selected. Copyright © 2014 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

The potential of pathogens as biological control of parthenium weed (Parthenium hysterophorus L.) in Ethiopia.

PubMed

Taye, T; Gossmann, M; Einhorn, G; Büttner, C; Metz, R; Abate, D

2002-01-01

P. hsyterophorus is an exotic invasive annual weed now causing severe infestation in Ethiopia. Studies on diagnosis, incidence and distribution of pathogens associated with parthenium weed in Ethiopia were carried out from 1998-2002. Several fungal isolates were obtained from seed and other parts of parthenium plants. Among them were putative pathogenic fungal species of the genus Helminthosporium, Phoma, Curvularia, Chaetomium, Alternaria, and Fusarium. However, pathogenecity test of the isolates obtained showed no or non-specific symptoms. It was concluded that these pathogens could be opportunistic with insignificant potential for biological control of parthenium. Two most important diseases associated with parthenium were a rust disease, caused by Puccinia abrupta var. partheniicola, and a phyllody disease, caused by a phytoplasma of fababean phyllody (PBP) phytoplasma group. The rust was commonly found in cool mid altitude (1500-2500 m) areas while phyllody was observed in low to mid altitude regions (900-2500 m) of Ethiopia, with a disease incidence up to 100% and 75%, respectively, in some locations. Study of the individual effects of the rust and phyllody diseases under field conditions showed a reduction on weed morphological parameters (plant height, leaf area, and dry matter yield). Parthenium seed production was reduced by 42% and 85% due to rust and phyllody, respectively. Phyllody and rust diseases of parthenium showed significant potential for classical biological control of parthenium after further confirmation of insect vectors that transmit phyllody and host range of phyllody disease to the related economic plants in Ethiopia.

Assessing the infection risk of enteropathogens from consumption of raw vegetables washed with contaminated water in Kathmandu Valley, Nepal.

PubMed

Shrestha, S; Haramoto, E; Shindo, J

2017-11-01

To assess diarrhoeal risks from enteropathogenic Escherichia coli, Giardia and Cryptosporidium from consuming raw spinach, cabbage, carrots and tomatoes in Kathmandu Valley, Nepal. The annual infection risk was quantified using the probabilistic Quantitative Microbial Risk Assessment approach, which considered 12 vegetable washing combinations. A new model was used to estimate dose of pathogens per exposure comprising parameters such as pathogen concentration in vegetable wash water before selling and eating, vegetable consumption rate, remaining pathogen ratio after washing, remaining water on vegetables after washing and water treatment removal efficiency. When all washing combinations were considered, high infection risks above the acceptable level of -4 log 10 infection per person per year were obtained, whereas the risk was reduced when other sources excluding river water were used. Assuming use of water treated with ceramic filters by all consumers, a 0-2 log 10 reduction in the estimated risks was obtained, which was insufficient to achieve the required risk level. High risk of diarrhoea prevails among raw vegetable consumers in the valley. It is needed to protect vegetable washing water sources and establish advanced water treatment methods to achieve the required level of public health risk. © 2017 The Society for Applied Microbiology.

Microbial minimalism: genome reduction in bacterial pathogens.

PubMed

Moran, Nancy A

2002-03-08

When bacterial lineages make the transition from free-living or facultatively parasitic life cycles to permanent associations with hosts, they undergo a major loss of genes and DNA. Complete genome sequences are providing an understanding of how extreme genome reduction affects evolutionary directions and metabolic capabilities of obligate pathogens and symbionts.

Improvement of resistance to rust through recurrent selection in pearl millet

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tapsoba, H.; Wilson, J.P.; Hanna, W.W.

Two pearl millet [Pennisetum glaucum (L.) R.Br. = P. typhoides (Burm.) Staff & Hubb., P. americanum (L.) K. Schum.] bulk populations, Tift No. 2 and Tift No. 5, served as base populations for four cycles of recurrent selection against susceptibility to Puccinia substriata Ell. & Barth, var. indica Ramachar & Cumm. A bulk inoculum of the pathogen was used. The objectives were to evaluate the progress achieved regarding overall resistance to the pathogen in the field and resistance to different races of the pathogen, and also to evaluate changes in unselected traits. During selection, the frequency of rust resistant plantsmore » continuously increased from about 30% in each base population to more than 85% by the third cycle of selection in both populations. An average increase of about 21 and 18% per cycle was obtained in Tift No. 2 and Tift No. 5, respectively. A continuous increase of the frequency of plants resistant to some races of the pathogen was also obtained. In Tift No. 5, 80% of the plants were resistant to eight races by the third cycle of selection. The accumulation of resistance observed in the seedlings was manifested in the field, both in 1993 and 1994, by a reduction of the final rust severity from the base population to the fourth selection cycle of both populations. This improvement in resistance to the rust pathogen was accompanied by an increase in the frequency of plants resistant to Pyricularia grisea (Cooke) Sacc. only in Tift No. 2. Despite the improvement in the selected character, genetic variability for agronomic traits such as plant height, number of culms/plant, flowering date, and panicle length was successfully maintained within each population. 20 refs., 1 fig., 7 tabs.« less

Bacterial contamination of stethoscope chest pieces and the effect of daily cleaning.

PubMed

Fujita, H; Hansen, B; Hanel, R

2013-01-01

Stethoscopes are a potential source of nosocomial infection for hospitalized humans, a phenomenon not previously studied in companion animals. To determine if daily cleaning of stethoscope chest pieces reduces bacterial contamination between cleanings. Client-owned dogs and cats. Prospective observational study. In phase 1, bacterial cultures were obtained from the chest pieces of 10 participant stethoscopes once weekly for 3 weeks. In phase 2, stethoscopes were cleaned daily and 2 culture samples were obtained once weekly, immediately before and after cleaning with 70% isopropyl alcohol, for 3 weeks. Daily cleaning eliminated bacteria immediately after each cleaning (P = .004), but did not reduce the rate of positive cultures obtained before cleaning in phase 2. Cultures were positive for 20/30 (67%) samples during phase 1 and 18/30 (60%) obtained before daily cleaning during phase 2. Recovered organisms included normal skin flora, agents of opportunistic infections, and potential pathogens. The only genus that was repeatedly recovered from the same stethoscope for 2 or more consecutive weeks was Bacillus sp. Daily cleaning was highly effective at removing bacteria, but provided no reduction in precleaning contamination. Cleaning stethoscopes after use on dogs or cats infected with pathogenic bacteria and before use on immunocompromised animals should be considered. Copyright © 2013 by the American College of Veterinary Internal Medicine.

Estimating the Effectiveness of Imidacloprid When Used to Suppress Transmission of Tomato spotted wilt orthotospovirus in Commercial Agriculture.

PubMed

Chappell, Thomas M; Kennedy, George G

2018-06-21

Imidacloprid is widely used to manage tomato spotted wilt disease (TSW) in tobacco, tomato, and pepper, caused by Tomato spotted wilt orthotospovirus (TSWV) and spread by the tobacco thrips, Frankliniella fusca Hinds (Thysanoptera: Thripidae). Imidacloprid suppresses transmission of TSWV by reducing probing and feeding by adult thrips on treated plants, thereby reducing the probability of transmission by infectious thrips. Because imidacloprid does not reduce probing and feeding on treated plants to zero, the reduction in transmission probability per viruliferous thrips can be offset by an increase in the number of viruliferous thrips challenging treated plants. A composite of these effects which we call 'pathogen pressure' experienced by plants is a function of thrips population size, the proportion of those thrips that are viruliferous, and the probability that viruliferous thrips successfully inoculate plants. To better understand the relationship between imidacloprid's effect on virus transmission, pathogen pressure, and TSW incidence in tobacco, we modeled TSW incidence as a function of the two most important variables affecting components of pathogen pressure, temperature, and precipitation, and the dependence of imidacloprid's effect on pathogen pressure. A model incorporating imidacloprid's effect as a reduction in pathogen pressure was found to be more descriptive than models incorporating the effect as a reduction in TSW incidence. Results reveal maximum proportional reduction in TSW incidence resulting from imidacloprid use is associated with minimal potential TSW incidence. As pathogen pressure increases, potential TSW incidence approaches 100%, and the benefits of imidacloprid use are highest at intermediate levels of pathogen pressure.

Risk-based enteric pathogen reduction targets for non-potable and direct potable use of roof runoff, stormwater, and greywater

EPA Science Inventory

This paper presents risk-based enteric pathogen log reduction targets for non-potable and potable uses of a variety of alternative source waters (i.e., locally-collected greywater, roof runoff, and stormwater). A probabilistic Quantitative Microbial Risk Assessment (QMRA) was use...

Pathogen inactivation in liquid dairy manure during anaerobic and aerobic digestions

NASA Astrophysics Data System (ADS)

Biswas, S.; Pandey, P.; Castillo, A. R.; Vaddella, V. K.

2014-12-01

Controlling manure-borne pathogens such as E. coli O157:H7, Salmonella spp. and Listeria monocytogenes are crucial for protecting surface and ground water as well as mitigating risks to human health. In California dairy farms, flushing of dairy manure (mainly animal feces and urine) from freestall barns and subsequent liquid-solid manure separation is a common practice for handling animal waste. The liquid manure fraction is generally pumped into the settling ponds and it goes into aerobic and/or anaerobic lagoons for extended period of time. Considering the importance of controlling pathogens in animal waste, the objective of the study was to understand the effects of anaerobic and aerobic digestions on the survival of three human pathogens in animal waste. The pathogen inactivation was assessed at four temperatures (30, 35, 42, and 50 °C), and the relationships between temperature and pathogen decay were estimated. Results showed a steady decrease of E. coli levels in aerobic and anaerobic digestion processes over the time; however, the decay rates varied with pathogens. The effect of temperature on Salmonella spp. and Listeria monocytogenes survival was different than the E. coli survival. In thermophilic temperatures (42 and 50 °C), decay rate was considerable greater compared to the mesophilic temperatures (30 and 35°C). The E. coli log reductions at 50 °C were 2.1 in both aerobic and anaerobic digestions after 13 days of incubation. The Salmonella spp. log reductions at 50 °C were 5.5 in aerobic digestion, and 5.9 in anaerobic digestion. The Listeria monocytogenes log reductions at 50 °C were 5.0 in aerobic digestion, and 5.6 in anaerobic digestion. The log reduction of E. coli, Salmonella spp., and Listeria monocytogens at 30 °C in aerobic environment were 0.1, 4.7, and 5.6, respectively. In anaerobic environment, the corresponding reductions were 0.4, 4.3, and 5.6, respectively. We anticipate that the outcomes of the study will help improving the existing animal waste management processes to control manure-borne pathogens.

Survival of Spoilage and Pathogenic Microorganisms on Cardboard and Plastic Packaging Materials

PubMed Central

Siroli, Lorenzo; Patrignani, Francesca; Serrazanetti, Diana I.; Chiavari, Cristiana; Benevelli, Marzia; Grazia, Luigi; Lanciotti, Rosalba

2017-01-01

The aim of this work was to study the interaction of corrugated and plastic materials with pathogenic and spoiling microorganisms frequently associated to fresh produce. The effect of the two packaging materials on the survival during the storage of microorganisms belonging to the species Escherichia coli, Listeria monocytogenes, Salmonella enteritidis, Saccharomyces cerevisiae, Lactobacillus plantarum, Pseudomonas fluorescens, and Aspergillus flavus was studied through traditional plate counting and scanning electron microscopy (SEM). The results obtained showed that cardboard materials, if correctly stored, reduced the potential of packaging to cross-contaminate food due to a faster viability loss by spoilage and pathogenic microorganisms compared to the plastic ones. In fact, the cell loads of the pathogenic species considered decreased over time independently on the inoculation level and packaging material used. However, the superficial viability losses were significantly faster in cardboard compared to plastic materials. The same behavior was observed for the spoilage microorganisms considered. The SEM microphotographs indicate that the reduction of superficial contamination on cardboard surfaces was due to the entrapping of the microbial cells within the fibers and the pores of this material. In addition, SEM data showed that the entrapped cells were subjected to more or less rapid lyses, depending on the species, due to the absence of water and nutrients, with the exception of molds. The latter spoilers were able to proliferate inside the cardboard fibers only when the absorption of water was not prevented during the storage. In conclusion, the findings of this work showed the reduction of cross-contamination potential of corrugated compared to plastic packaging materials used in fruit and vegetable sector. However, the findings outlined the importance of hygiene and low humidity during cardboard storage to prevent the mold growth on packaging. PMID:29312271

Changes in the fungal autoflora of Apollo astronauts.

PubMed

Taylor, G R; Henney, M R; Ellis, W L

1973-11-01

Specimens were repeatedly obtained for mycological examination from the skin, throat, urine, and feces of the six astronauts who conducted the Apollo 14 and Apollo 15 lunar exploration missions. Analysis of preflight data demonstrates that the process of severely restricting opportunities from colonization for 3 weeks before flight resulted in a 50% reduction in the number of isolated species. Postflight data indicate that exposure to the space flight environment for up to 2 weeks resulted in an even greater reduction with a relative increase in the potential pathogen Candida albicans. No incidences of microbial shock were observed when crewmembers were quarantined for 16 days after completion of the space flight. Intercrew transfer of particular species could not be demonstrated because most species were not consistently recovered.

Ability of Nonpathogenic Fusarium oxysporum Strain Fo47 To Induce Resistance against Pythium ultimum Infection in Cucumber

PubMed Central

Benhamou, Nicole; Garand, Chantal; Goulet, Alain

2002-01-01

The influence exerted by nonpathogenic Fusarium oxysporum strain Fo47 in triggering cucumber protection against infection by Pythium ultimum was investigated ultrastructurally. Macroscopic and microscopic observations of the pathogen colony in dual cultures revealed that reduction of Pythium growth was associated with marked disorders, including generalized disorganization of the host cytoplasm, retraction of the plasmalemma, and complete loss of the protoplasm. Cytochemical labeling of cellulose with an exoglucanase-gold complex showed that the cellulose component of the host cell walls was structurally preserved at a time when the host cytoplasm had undergone complete disorganization. A similar antagonistic process was observed at the root cell surface. Most striking and interesting was the finding that mycoparasitism, as evidenced by the frequent occurrence of Fo47 hyphae within nearly empty cells of the pathogen, occurred not only at the root surface but also within the invaded root tissues. The specific labeling pattern obtained with the exoglucanase-gold complex confirmed that Fo47 successfully penetrated cells of the pathogen, both in the rhizosphere and inside the root tissues. Pythium cells that could evade the first defensive line in the rhizosphere could penetrate the root epidermis, but their growth was restricted to the outermost tissues. Positive correlations between Fo47 treatment and induced resistance to infection by P. ultimum in cucumber were confirmed by (i) the reduction of pathogen viability; (ii) the elaboration of newly formed barriers, a phenomenon which was not seen in Fo47-free plants, where the pathogen proliferated in all root tissues within a few days; and (iii) the occlusion of intercellular spaces with a dense material likely enriched in phenolics. Taken together, our observations provide the first convincing evidence that Fo47 exerts a direct inhibitory effect on P. ultimum through a combination of antibiosis and mycoparasitism, in addition to being a strong inducer of plant defense reactions. PMID:12147506

Assessment of bacterial and archaeal community structure in Swine wastewater treatment processes.

PubMed

Da Silva, Marcio Luis Busi; Cantão, Mauricio Egídio; Mezzari, Melissa Paola; Ma, Jie; Nossa, Carlos Wolfgang

2015-07-01

Microbial communities from two field-scale swine wastewater treatment plants (WWTPs) were assessed by pyrosequencing analyses of bacterial and archaeal 16S ribosomal DNA (rDNA) fragments. Effluent samples from secondary (anaerobic covered lagoons and upflow anaerobic sludge blanket [UASB]) and tertiary treatment systems (open-pond natural attenuation lagoon and air-sparged nitrification-denitrification tank followed by alkaline phosphorus precipitation process) were analyzed. A total of 56,807 and 48,859 high-quality reads were obtained from bacterial and archaeal libraries, respectively. Dominant bacterial communities were associated with the phylum Firmicutes, Bacteroidetes, Proteobacteria, or Actinobacteria. Bacteria and archaea diversity were highest in UASB effluent sample. Escherichia, Lactobacillus, Bacteroides, and/or Prevotella were used as indicators of putative pathogen reduction throughout the WWTPs. Satisfactory pathogen reduction was observed after the open-pond natural attenuation lagoon but not after the air-sparged nitrification/denitrification followed by alkaline phosphorus precipitation treatment processes. Among the archaeal communities, 80% of the reads was related to hydrogeno-trophic methanogens Methanospirillum. Enrichment of hydrogenotrophic methanogens detected in effluent samples from the anaerobic covered lagoons and UASB suggested that CO2 reduction with H2 was the dominant methanogenic pathway in these systems. Overall, the results served to improve our current understanding of major microbial communities' changes downgradient from the pen and throughout swine WWTP as a result of different treatment processes.

The RpfCG two-component system negatively regulates the colonization of sugar cane stalks by Xanthomonas albilineans.

PubMed

Rott, Philippe; Fleites, Laura A; Mensi, Imène; Sheppard, Lauren; Daugrois, Jean-Heinrich; Dow, J Maxwell; Gabriel, Dean W

2013-06-01

The genome of Xanthomonas albilineans, the causal agent of sugar cane leaf scald, carries a gene cluster encoding a predicted quorum sensing system that is highly related to the diffusible signalling factor (DSF) systems of the plant pathogens Xylella fastidiosa and Xanthomonas campestris. In these latter pathogens, a cluster of regulation of pathogenicity factors (rpf) genes encodes the DSF system and is involved in control of various cellular processes. Mutation of Xanthomonas albilineans rpfF, encoding a predicted DSF synthase, in Florida strain XaFL07-1 resulted in a small reduction of disease severity (DS). Single-knockout mutations of rpfC and rpfG (encoding a predicted DSF sensor and regulator, respectively) had no effect on DS or swimming motility of the pathogen. However, capacity of the pathogen to cause disease was slightly reduced and swimming motility was severely affected when rpfG and rpfC were both deleted. Similar results were obtained when the entire rpfGCF region was deleted. Surprisingly, when the pathogen was mutated in rpfG or rpfC (single or double mutations) it was able to colonize sugar cane spatially more efficiently than the wild-type. Mutation in rpfF alone did not affect the degree of spatial invasion. We conclude that the DSF signal contributes to symptom expression but not to invasion of sugar cane stalks by Xanthomonas albilineans strain XaFL07-1, which is mainly controlled by the RpfCG two-component system.

TREATMENT OF MUNICIPAL SLUDGE FOR PATHOGEN REDUCTION

EPA Science Inventory

This presentation reviews the pathogenic microorganisms that may be found in municipal sewage sludge and the commonly employed Class A and B processes for controlling pathogens. It notes how extensively they are used and discusses issues and concerns with their application. The...

The effect of tar spot pathogen on host plant carbon balance and its possible consequences on a tundra ecosystem.

PubMed

Masumoto, Shota; Uchida, Masaki; Tojo, Motoaki; Herrero, Maria Luz; Mori, Akira S; Imura, Satoshi

2018-03-01

In Arctic tundra, plant pathogens have substantial effects on the growth and survival of hosts, and impacts on the carbon balance at the scale of ecological systems. To understand these effects on carbon dynamics across different scales including plant organ, individual, population and ecosystem, we focused on two primary factors: host productivity reduction and carbon consumption by the pathogen. We measured the effect of the pathogen on photosynthetic and respiratory activity in the host. We also measured respiration and the amount of carbon in the pathogen. We constructed a model based on these two factors, and calculated pathogenic effects on the carbon balance at different organismal and ecological scales. We found that carbon was reduced in infected leaves by 118% compared with healthy leaves; the major factor causing this loss was pathogenic carbon consumption. The carbon balance at the population and ecosystem levels decreased by 35% and 20%, respectively, at an infection rate of 30%. This case study provides the first evidence that a host plant can lose more carbon through pathogenic carbon consumption than through a reduction in productivity. Such a pathogenic effect could greatly change ecosystem carbon cycling without decreasing annual productivity.

The effectiveness of riboflavin and ultraviolet light pathogen reduction technology in eliminating Trypanosoma cruzi from leukoreduced whole blood.

PubMed

Jimenez-Marco, Teresa; Cancino-Faure, Beatriz; Girona-Llobera, Enrique; Alcover, M Magdalena; Riera, Cristina; Fisa, Roser

2017-06-01

The parasitic Chagas disease is caused by the protozoan Trypanosoma cruzi, which is mainly transmitted by insect vectors. Other infection routes, both in endemic and in nonendemic areas, include organ and marrow transplantation, congenital transmission, and blood transfusion. Asymptomatic chronic chagasic individuals may have a low and transient parasitemia in peripheral blood and, consequently, they can unknowingly transmit the disease via blood transfusion. Riboflavin and ultraviolet (UV) light pathogen reduction is a method to reduce pathogen transfusion transmission risk based on damage to the pathogen nucleic acids. In this study, we tested the effectiveness of this technology for the elimination of T. cruzi parasites in artificially contaminated whole blood units (WBUs) and thus for decreasing the risk of T. cruzi transfusion transmission. The contaminated WBUs were leukoreduced by filtration and treated with riboflavin and UV light. The level of pathogen reduction was quantified by a real-time polymerase chain reaction (qPCR) and a real-time reverse transcription-polymerase chain reaction (RT-qPCR) as a viability assay. The RNA (cDNA) quantification of the parasites showed a more than 99% reduction of viable T. cruzi parasites after leukoreduction and a complete reduction (100%) after the riboflavin and UV light treatment. Riboflavin and UV light treatment and leukoreduction used in conjunction appears to eliminate significant amounts of viable T. cruzi in whole blood. Both strategies could complement other blood bank measures already implemented to prevent the transmission of T. cruzi via blood transfusion. © 2017 AABB.

Whole-leaf wash improves chlorine efficacy for microbial reduction and prevents pathogen cross-contamination during fresh-cut lettuce processing.

PubMed

Nou, Xiangwu; Luo, Yaguang

2010-06-01

Currently, most fresh-cut processing facilities in the United States use chlorinated water or other sanitizer solutions for microbial reduction after lettuce is cut. Freshly cut lettuce releases significant amounts of organic matter that negatively impacts the effectiveness of chlorine or other sanitizers for microbial reduction. The objective of this study is to evaluate whether a sanitizer wash before cutting improves microbial reduction efficacy compared to a traditional postcutting sanitizer wash. Romaine lettuce leaves were quantitatively inoculated with E. coli O157:H7 strains and washed in chlorinated water before or after cutting, and E. coli O157:H7 cells that survived the washing process were enumerated to determine the effectiveness of microbial reduction for the 2 cutting and washing sequences. Whole-leaf washing in chlorinated water improved pathogen reduction by approximately 1 log unit over traditional cut-leaf sanitization. Similar improvement in the reduction of background microflora was also observed. Inoculated "Lollo Rossa" red lettuce leaves were mixed with noninoculated Green-Leaf lettuce leaves to evaluate pathogen cross-contamination during processing. High level (96.7% subsamples, average MPN 0.6 log CFU/g) of cross-contamination of noninoculated green leaves by inoculated red leaves was observed when mixed lettuce leaves were cut prior to washing in chlorinated water. In contrast, cross-contamination of noninoculated green leaves was significantly reduced (3.3% of subsamples, average MPN

Pathogen reduction of blood components.

PubMed

Solheim, Bjarte G

2008-08-01

Thanks to many blood safety interventions introduced in developed countries the risk of transfusion transmitted infections has become exceedingly small in these countries. However, emerging pathogens still represent a serious challenge, as demonstrated by West Nile virus in the US and more recently by Chikungunya virus in the Indian Ocean. In addition bacterial contamination, particularly in platelets, and protozoa transmitted by blood components still represent sizeable risks in developed countries. In developing countries the risk of all transfusion transmitted infections is still high due to insufficient funding and organisation of the health service. Pathogen reduction of pooled plasma products has virtually eliminated the risk of transfusion transmitted infections, without compromising the quality of the products significantly. Pathogen reduction of blood components has been much more challenging. Solvent detergent treatment which has been so successfully applied for plasma products dissolves cell membranes, and can, therefore, only be applied for plasma and not for cellular blood components. Targeting of nucleic acids has been another method for pathogen inactivation of plasma and the only approach possible for cellular blood products. As documented in more than 15 year's track record, solvent detergent treatment of pooled plasma can yield high quality plasma. The increased risk for contamination by unknown viruses due to pooling is out weighed by elimination of TRALI, significant reduction in allergic reactions and standardisation of the product. Recently, a promising method for solvent detergent treatment of single donor plasma units has been published. Methylene blue light treatment of single donor plasma units has a similar long track record as pooled solvent detergent treated plasma; but the method is less well documented and affects coagulation factor activity more. Psoralen light treated plasma has only recently been introduced (CE marked in Europe, but not licensed by the FDA), while the method of Riboflavin light treatment of plasma still is under development. In addition to pathogen reduction the methods, however, result in some reduction of coagulation factor activity. For platelets only Psoralen and Riboflavin light treatment have been implemented. Both are CE marked products in Europe but only approved for clinical trials in the USA. The methods affect platelet activity, but result in clinically acceptable platelets with only slightly reduced CCI and increased demand for platelet transfusions. Pathogen reduction of red blood cells with FRALE (S-303) or INACTINE (PEN110) has so far resulted in the formation of antibodies against neo-epitopes on red blood cells. A promising method for Riboflavin treatment of red blood cells is under development. This manuscript reviews the current experience and discusses future trends.

Changes in the Fungal Autoflora of Apollo Astronauts

PubMed Central

Taylor, Gerald R.; Henney, Mary R.; Ellis, Walter L.

1973-01-01

Specimens were repeatedly obtained for mycological examination from the skin, throat, urine, and feces of the six astronauts who conducted the Apollo 14 and Apollo 15 lunar exploration missions. Analysis of preflight data demonstrates that the process of severely restricting opportunities from colonization for 3 weeks before flight resulted in a 50% reduction in the number of isolated species. Postflight data indicate that exposure to the space flight environment for up to 2 weeks resulted in an even greater reduction with a relative increase in the potential pathogen Candida albicans. No incidences of microbial shock were observed when crewmembers were quarantined for 16 days after completion of the space flight. Intercrew transfer of particular species could not be demonstrated because most species were not consistently recovered. PMID:4762399

Siderophore-mediated iron acquisition mechanisms in Vibrio vulnificus biotype 2.

PubMed Central

Biosca, E G; Fouz, B; Alcaide, E; Amaro, C

1996-01-01

Vibrio vulnificus biotype 2 is a primary pathogen for eels and, as has recently been suggested, an opportunistic pathogen for humans. In this study we have investigated the ability of V. vulnificus biotype 2 to obtain iron by siderophore-mediated mechanisms and evaluated the importance of free iron in vibriosis. The virulence degree for eels was dependent on iron availability from host fluids, as was revealed by a reduction in the 50% lethal dose for iron-overloaded eels. This biotype produced both phenolate- and hydroxamate-type siderophores of an unknown nature and two new outer membrane proteins of around 84 and 72 kDa in response to iron starvation. No alterations in lipopolysaccharide patterns were detected in response to iron stress. Finally, our data suggest that V. vulnificus biotype 2 uses the hydroxamate-type siderophore for removal of iron from transferrin rather than relying on a receptor for this iron-binding protein. PMID:8975620

Viable-but-Nonculturable Listeria monocytogenes and Salmonella enterica Serovar Thompson Induced by Chlorine Stress Remain Infectious.

PubMed

Highmore, Callum J; Warner, Jennifer C; Rothwell, Steve D; Wilks, Sandra A; Keevil, C William

2018-04-17

The microbiological safety of fresh produce is monitored almost exclusively by culture-based detection methods. However, bacterial food-borne pathogens are known to enter a viable-but-nonculturable (VBNC) state in response to environmental stresses such as chlorine, which is commonly used for fresh produce decontamination. Here, complete VBNC induction of green fluorescent protein-tagged Listeria monocytogenes and Salmonella enterica serovar Thompson was achieved by exposure to 12 and 3 ppm chlorine, respectively. The pathogens were subjected to chlorine washing following incubation on spinach leaves. Culture data revealed that total viable L. monocytogenes and Salmonella Thompson populations became VBNC by 50 and 100 ppm chlorine, respectively, while enumeration by direct viable counting found that chlorine caused a <1-log reduction in viability. The pathogenicity of chlorine-induced VBNC L. monocytogenes and Salmonella Thompson was assessed by using Caenorhabditis elegans Ingestion of VBNC pathogens by C. elegans resulted in a significant life span reduction ( P = 0.0064 and P < 0.0001), and no significant difference between the life span reductions caused by the VBNC and culturable L. monocytogenes treatments was observed. L. monocytogenes was visualized beyond the nematode intestinal lumen, indicating resuscitation and cell invasion. These data emphasize the risk that VBNC food-borne pathogens could pose to public health should they continue to go undetected. IMPORTANCE Many bacteria are known to enter a viable-but-nonculturable (VBNC) state in response to environmental stresses. VBNC cells cannot be detected by standard laboratory culture techniques, presenting a problem for the food industry, which uses these techniques to detect pathogen contaminants. This study found that chlorine, a sanitizer commonly used for fresh produce, induces a VBNC state in the food-borne pathogens Listeria monocytogenes and Salmonella enterica It was also found that chlorine is ineffective at killing total populations of the pathogens. A life span reduction was observed in Caenorhabditis elegans that ingested these VBNC pathogens, with VBNC L. monocytogenes as infectious as its culturable counterpart. These data show that VBNC food-borne pathogens can both be generated and avoid detection by industrial practices while potentially retaining the ability to cause disease. Copyright © 2018 Highmore et al.

Pathogen reduction in minimally managed composting of bovine manure

USDA-ARS?s Scientific Manuscript database

Persistence of pathogenic bacteria such as E. coli O157:H7, Salmonella spp., and Listeria monocytogenes in bovine feces and contaminated soils is an important risk factor in perpetuating the initial infection as well as re-infection of cattle and dissemination of pathogens throughout agricultural la...

FROM CONCEPT TO EQUIVALENCY: GETTING YOUR INNOVATIVE SLUDGE DISINFECTION PROCESS CLASSIFIED AS A OR B IS NOW LESS OF A MYSTERY

EPA Science Inventory

The Pathogen Equivalency Committee has updated the criteria it uses to make recommendations of equivalency on innovative or alternative sludge pathogen reduction processes. To assist new applicants through the equivalency recommendation process the pathogen equivalency committee ...

Pasteurization of milk: the heat inactivation kinetics of milk-borne dairy pathogens under commercial-type conditions of turbulent flow.

PubMed

Pearce, L E; Smythe, B W; Crawford, R A; Oakley, E; Hathaway, S C; Shepherd, J M

2012-01-01

This is the first study to report kinetic data on the survival of a range of significant milk-borne pathogens under commercial-type pasteurization conditions. The most heat-resistant strain of each of the milk-borne pathogens Staphylococcus aureus, Yersinia enterocolitica, pathogenic Escherichia coli, Cronobacter sakazakii (formerly known as Enterobacter sakazakii), Listeria monocytogenes, and Salmonella was selected to obtain the worst-case scenario in heat inactivation trials using a pilot-plant-scale pasteurizer. Initially, approximately 30 of each species were screened using a submerged coil unit. Then, UHT milk was inoculated with the most heat-resistant pathogens at ~10(7)/mL and heat treated in a pilot-plant-scale pasteurizer under commercial-type conditions of turbulent flow for 15s over a temperature range from 56 to 66°C and at 72°C. Survivors were enumerated on nonselective media chosen for the highest efficiency of plating of heat-damaged bacteria of each of the chosen strains. The mean log(10) reductions and temperatures of inactivation of the 6 pathogens during a 15-s treatment were Staph. aureus >6.7 at 66.5°C, Y. enterocolitica >6.8 at 62.5°C, pathogenic E. coli >6.8 at 65°C, C. sakazakii >6.7 at 67.5°C, L. monocytogenes >6.9 at 65.5°C, and Salmonella ser. Typhimurium >6.9 at 61.5°C. The kinetic data from these experiments will be used by the New Zealand Ministry of Agriculture and Forestry to populate the quantitative risk assessment model being developed to investigate the risks to New Zealand consumers from pasteurized, compared with nonpasteurized, milk and milk products. Copyright © 2012 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Comparison of antifungal activities of various essential oils on the Phytophthora drechsleri, the causal agent of fruit decay

PubMed Central

Mohammadi, Ali; Hashemi, Maryam; Hosseini, Seyed Masoud

2015-01-01

Background and Objectives: The efficacy of Mentha piperita L, Zataria multiflora Boiss and Thymus vulgaris L essential oils (EOs) was evaluated for controlling the growth of Phytophthora drechsleri, the causative agent of damage to many crops that is consumed directly by humans. Materials and Methods: The EOs used in this study was purchased from Magnolia Co, Iran. The pour plate method in petri dishes containing Potato Dextrose Agar (PDA) was used to evaluate the antifungal properties of EOs. The minimal inhibitory concentrations (MIC), minimum fungicidal concentration (MFC) as well as mycelial growth inhibition (MGI) were measured. The IC50 value (the concentration inhibited 50% of the mycelium growth) was calculated by probit analysis. Results and Conclusion: The fungal growth was significantly reduced by increasing concentrations of tested EOs. The complete reduction was obtained with Shirazi thyme at all concentrations, whereas the complete reduction for peppermint and thyme was observed at 0.4% and 0.8% (v/v) concentrations, respectively. Meanwhile, the minimum inhibition was observed when 0.1% peppermint (MGI values of 9.37%) was used. The IC50, MIC and MFC values of Shirazi thyme was 0.053, 0.1% and 0.2%, respectively. Similarly, MIC and MFC values of peppermint and thyme were recorded 0.4% and 0.8%, respectively. The results obtained from this study may contribute to the development of new antifungal agents to protect the crops from this pathogenic fungus and many agricultural plant pathogens causing drastic crop losses. PMID:26644871

Pathogen Treatment Guidance and Monitoring Approaches fro ...

EPA Pesticide Factsheets

On-site non-potable water reuse is increasingly used to augment water supplies, but traditional fecal indicator approaches for defining and monitoring exposure risks are limited when applied to these decentralized options. This session emphasizes risk-based modeling to define pathogen log-reduction requirements coupled with alternative targets for monitoring enabled by genomic sequencing (i.e., the microbiome of reuse systems). 1. Discuss risk-based modeling to define pathogen log-reduction requirements 2. Review alternative targets for monitoring 3. Gain an understanding of how new tools can help improve successful development of sustainable on-site non-potable water reuse Presented at the Water Wastewater Equipment Treatment & Transport Show.

A transient expression assay for the in planta efficacy screening of an antimicrobial peptide against grapevine bacterial pathogens.

PubMed

Visser, M; Stephan, D; Jaynes, J M; Burger, J T

2012-06-01

Natural and synthetic antimicrobial peptides (AMPs) are of increasing interest as potential resistance conferring elements in plants against pathogen infection. The efficacy of AMPs against pathogens is prescreened by in vitro assays, and promising AMP candidates are introduced as transgenes into plants. As in vitro and in planta environments differ, a prescreening procedure of the AMP efficacy in the plant environment is desired. Here, we report the efficacy of the purified synthetic peptide D4E1 against the grapevine-infecting bacterial pathogens Agrobacterium vitis and Xylophilus ampelinus in vitro and describe for the first time an in planta prescreening procedure based on transiently expressed D4E1. The antimicrobial effect of D4E1 against Ag. vitis and X. ampelinus was shown by a reduction in colony-forming units in vitro in a traditional plate-based assay and by a reduction in bacterial titres in planta as measured by quantitative real-time PCR (qPCR) in grapevine leaves transiently expressing D4E1. A statistically significant reduction in titre was shown for X. ampelinus, but for Ag. vitis, a significant reduction in titre was only observed in a subset of plants. The titres of both grapevine-infecting bacterial pathogens were reduced in an in vitro assay and for X. ampelinus in an in planta assay by D4E1 application. This widens the applicability of D4E1 as a potential resistance-enhancing element to additional pathogens and in a novel plant species. D4E1 is a promising candidate to confer enhanced resistance against the two tested grapevine bacterial pathogens, and the applied transient expression system proved to be a valuable tool for prescreening of D4E1 efficacy in an in planta environment. The described prescreening procedure can be used for other AMPs and might be adapted to other plant species and pathogens before the expensive and tedious development of stably transgenic lines is started. © 2012 The Authors. Letters in Applied Microbiology © 2012 The Society for Applied Microbiology.

Development of a process-based model to predict pathogen budgets for the Sydney drinking water catchment.

PubMed

Ferguson, Christobel M; Croke, Barry F W; Beatson, Peter J; Ashbolt, Nicholas J; Deere, Daniel A

2007-06-01

In drinking water catchments, reduction of pathogen loads delivered to reservoirs is an important priority for the management of raw source water quality. To assist with the evaluation of management options, a process-based mathematical model (pathogen catchment budgets - PCB) is developed to predict Cryptosporidium, Giardia and E. coli loads generated within and exported from drinking water catchments. The model quantifies the key processes affecting the generation and transport of microorganisms from humans and animals using land use and flow data, and catchment specific information including point sources such as sewage treatment plants and on-site systems. The resultant pathogen catchment budgets (PCB) can be used to prioritize the implementation of control measures for the reduction of pathogen risks to drinking water. The model is applied in the Wingecarribee catchment and used to rank those sub-catchments that would contribute the highest pathogen loads in dry weather, and in intermediate and large wet weather events. A sensitivity analysis of the model identifies that pathogen excretion rates from animals and humans, and manure mobilization rates are significant factors determining the output of the model and thus warrant further investigation.

QPCR Analysis of Total and Viable Enterococci in Waste Water Treatment: Comparison with Culture in Predicting Pathogen Reductions

EPA Science Inventory

BEACH Act amendment to Clean Water Act requires EPA to establish more expeditious methods for the timely detection of pathogens and pathogen indicators in coastal waters New methods should demonstrate utility for and be compatible with all CWA 304(a) criteria needs including:...

Biocontrol of Botrytis cinerea and Calonectria gracilis by eucalypts growth promoters Bacillus spp.

PubMed

Paz, Isabel Cristina Padula; Santin, Rita de Cássia Madail; Guimarães, Alexandre Martins; Rosa, Osmar Paulo Pereira da; Quecine, Maria Carolina; Silva, Michele de Cássia Pereira E; Azevedo, João Lúcio; Matsumura, Aida Terezinha Santos

2018-05-17

The clonal Eucalyptus plants are commonly obtained by vegetative propagation under a protected environment. This system improves the Botrytis cinerea and Calonectria spp infection on the young eucalypts plantings, resulting gray mold and cutting rot respectively. Currently, the unique available control method is based on chemicals. As alternative, novel methods to manage plant diseases, endophytic microorganisms could be an interesting alternative. Thus, we aimed to evaluate endophytic Bacillus isolated from eucalypts as a biocontrol agent against Botrytis cinerea and Calonectria gracilis, important fungal pathogens in the greenhouse, using clonal plantlets of E. urograndis. Eight endophytic strains of Bacillus, previously described as eucalyptus growth promoters, were evaluated in vitro and in vivo against Botrytis cinerea and Calonectria gracilis. The diffusible metabolites assay showed the potential of endophytic Bacillus to decrease the growth of both pathogens. Differences in the susceptibility of the pathogens to bacterial volatile metabolites were observed, B. cinerea showed more susceptible than Calonectria gracilis. In vivo assays, Bacillus amyloliquefaciens EUCB 10 demonstrated better overall reductions in these diseases. Based on the results obtained from the in vitro and in vivo analyses, we suggest that the endophytic B. amyloliquefaciens strain EUCB 10 constitutes a promising biocontrol agent against B. cinerea and Calonectria gracilis. Furthermore, this is the first reporting of B. amyloliquefaciens previously describe as plant growth promoter and also as potential control agent of B. cinerea and Calonectria gracilis to eucalyptus. Copyright © 2018 Elsevier Ltd. All rights reserved.

Effect of Commercial Cyanobacteria Products on the Growth and Antagonistic Ability of Some Bioagents under Laboratory Conditions

PubMed Central

El-Mougy, Nehal S.; Abdel-Kader, Mokhtar M.

2013-01-01

Evaluation of the efficacy of blue-green algal compounds against the growth of either pathogenic or antagonistic microorganisms as well as their effect on the antagonistic ability of bioagents was studied under in vitro conditions. The present study was undertaken to explore the inhibitory effect of commercial algal compounds, Weed-Max and Oligo-Mix, against some soil-borne pathogens. In growth medium supplemented with these algal compounds, the linear growth of pathogenic fungi decreased by increasing tested concentrations of the two algal compounds. Complete reduction in pathogenic fungal growth was observed at 2% of both Weed-Max and Oligo-Mix. Gradual significant reduction in the pathogenic fungal growth was caused by the two bioagents and by increasing the concentrations of algal compounds Weed-Max and Oligo-Mix. The present work showed that commercial algal compounds, Weed-Max and Oligo-Mix, have potential for the suppression of soil-borne fungi and enhance the antagonistic ability of fungal, bacterial, and yeast bio-agents. PMID:24307948

Antimicrobial activity of lactic acid bacteria against Listeria monocytogenes on frankfurters formulated with and without lactate/diacetate.

PubMed

Koo, Ok-Kyung; Eggleton, Mallory; O'Bryan, Corliss A; Crandall, Philip G; Ricke, Steven C

2012-12-01

Contamination by Listeria monocytogenes has been a constant public health threat for the ready-to-eat (RTE) meat industry due to the potential for high mortalities from listeriosis. Lactic acid bacteria (LAB) have shown protective action against various pathogenic bacteria. The aim of this study was to evaluate the antilisterial activity of a combination of three LAB strains (Lactiguard®) on L. monocytogenes. The combination of the LAB was inhibitory to L. monocytogenes inoculated onto frankfurters not containing lactate/diacetate after 8weeks of refrigerated storage (0.6 log reduction compared to L. monocytogenes only control), and when a cell free extract (CFS) of the LAB was added with LAB even more inhibition was obtained (1.2 log reduction compared with L. monocytogenes only). In frankfurters containing lactate/diacetate the LAB and the LAB plus CFS were more effective in reducing growth of L. monocytogenes after 8 weeks of refrigerated storage (2 and 3.3 log reductions respectively). Copyright © 2012 Elsevier Ltd. All rights reserved.

Effect of in-house chicken litter composting on ammonia and nitrous oxide emissions and pathogen reduction

USDA-ARS?s Scientific Manuscript database

Inhouse composting is a management practice to reduce pathogen in poultry litter. In between flocks, growers windrow the litter inside the broiler houses. This results in high temperatures that can reduce some pathogens in the litter. However, this practice is likely to increase emissions of NH3 and...

A comprehensive proteomics study on platelet concentrates: Platelet proteome, storage time and Mirasol pathogen reduction technology.

PubMed

Salunkhe, Vishal; De Cuyper, Iris M; Papadopoulos, Petros; van der Meer, Pieter F; Daal, Brunette B; Villa-Fajardo, María; de Korte, Dirk; van den Berg, Timo K; Gutiérrez, Laura

2018-03-19

Platelet concentrates (PCs) represent a blood transfusion product with a major concern for safety as their storage temperature (20-24°C) allows bacterial growth, and their maximum storage time period (less than a week) precludes complete microbiological testing. Pathogen inactivation technologies (PITs) provide an additional layer of safety to the blood transfusion products from known and unknown pathogens such as bacteria, viruses, and parasites. In this context, PITs, such as Mirasol Pathogen Reduction Technology (PRT), have been developed and are implemented in many countries. However, several studies have shown in vitro that Mirasol PRT induces a certain level of platelet shape change, hyperactivation, basal degranulation, and increased oxidative damage during storage. It has been suggested that Mirasol PRT might accelerate what has been described as the platelet storage lesion (PSL), but supportive molecular signatures have not been obtained. We aimed at dissecting the influence of both variables, that is, Mirasol PRT and storage time, at the proteome level. We present comprehensive proteomics data analysis of Control PCs and PCs treated with Mirasol PRT at storage days 1, 2, 6, and 8. Our workflow was set to perform proteomics analysis using a gel-free and label-free quantification (LFQ) approach. Semi-quantification was based on LFQ signal intensities of identified proteins using MaxQuant/Perseus software platform. Data are available via ProteomeXchange with identifier PXD008119. We identified marginal differences between Mirasol PRT and Control PCs during storage. However, those significant changes at the proteome level were specifically related to the functional aspects previously described to affect platelets upon Mirasol PRT. In addition, the effect of Mirasol PRT on the platelet proteome appeared not to be exclusively due to an accelerated or enhanced PSL. In summary, semi-quantitative proteomics allows to discern between proteome changes due to Mirasol PRT or PSL, and proves to be a methodology suitable to phenotype platelets in an unbiased manner, in various physiological contexts.

Food decontamination using nanomaterials

USDA-ARS?s Scientific Manuscript database

The research indicates that nanomaterials including nanoemulsions are promising decontamination media for the reduction of food contaminating pathogens. The inhibitory effect of nanoparticles for pathogens could be due to deactivate cellular enzymes and DNA; disrupting of membrane permeability; and/...

Food Safety Impacts from Post-Harvest Processing Procedures of Molluscan Shellfish.

PubMed

Baker, George L

2016-04-18

Post-harvest Processing (PHP) methods are viable food processing methods employed to reduce human pathogens in molluscan shellfish that would normally be consumed raw, such as raw oysters on the half-shell. Efficacy of human pathogen reduction associated with PHP varies with respect to time, temperature, salinity, pressure, and process exposure. Regulatory requirements and PHP molluscan shellfish quality implications are major considerations for PHP usage. Food safety impacts associated with PHP of molluscan shellfish vary in their efficacy and may have synergistic outcomes when combined. Further research for many PHP methods are necessary and emerging PHP methods that result in minimal quality loss and effective human pathogen reduction should be explored.

Antimicrobial activity of broccoli (Brassica oleracea var. italica) cultivar Avenger against pathogenic bacteria, phytopathogenic filamentous fungi and yeast.

PubMed

Pacheco-Cano, R D; Salcedo-Hernández, R; López-Meza, J E; Bideshi, D K; Barboza-Corona, J E

2018-01-01

The objective of this study was to show whether the edible part of broccoli has antibacterial and antifungal activity against micro-organism of importance in human health and vegetable spoilage, and to test if this effect was partially due to antimicrobial peptides (AMPs). Crude extracts were obtained from florets and stems of broccoli cultivar Avenger and the inhibitory effect was demonstrated against pathogenic bacteria (Bacillus cereus, Staphylococcus xylosus, Staphylococcus aureus, Shigella flexneri, Shigella sonnei, Proteus vulgaris), phytopathogenic fungi (Colletotrichum gloeosporioides, Asperigillus niger) and yeasts (Candida albicans and Rhodotorula sp.). It was shown that samples treated with proteolytic enzymes had a reduction of approximately 60% in antibacterial activity against Staph. xylosus, suggesting that proteinaceous compounds might play a role in the inhibitory effect. Antimicrobial components in crude extracts were thermoresistant and the highest activity was observed under acidic conditions. It was shown that antifungal activity of broccoli's crude extracts might not be attributed to chitinases. Organic broccoli cultivar Avenger has antimicrobial activity against pathogenic bacteria, yeast and phytophatogenic fungi. Data suggest that this effect is partially due to AMPs. Broccoli's crude extracts have activity not only against pathogenic bacteria but also against phytophatogenic fungi of importance in agriculture. We suggest for first time that the inhibitory effect is probably due to AMPs. © 2017 The Society for Applied Microbiology.

MANAGING MICROBIAL CONTAMINATION IN URBAN WATERSHEDS

EPA Science Inventory

This paper presents different approaches for controlling pathogen contamination in urban watersheds for contamination resulting from point and diffuse sources. Point sources of pathogens can be treated by a disinfection technology of known effectiveness, and a desired reduction ...

MANAGING MICROBIAL CONTAMINATION IN URBAN WATERSHEDS

EPA Science Inventory

This paper presents different approaches for controlling pathogen contamination in urban watersheds for contamination resulting from point and diffuses sources. Point sources of pathogens can be treated by a disinfection technology of known effectiveness, and a desired reduction ...

Reduced MHC Alloimmunization and Partial Tolerance Protection With Pathogen Reduction Of Whole Blood

PubMed Central

Jackman, Rachael P.; Muench, Marcus O.; Inglis, Heather; Heitman, John W.; Marschner, Susanne; Goodrich, Raymond P.; Norris, Philip J.

2017-01-01

BACKGROUND Allogeneic blood transfusion can result in an immune response against major histocompatibility complex (MHC) antigens, potentially complicating future transfusions or transplants. We have previously shown that pathogen reduction of platelet-rich plasma (PRP) with riboflavin and UV light (UV+R) can prevent alloimmunization in mice. A similar pathogen reduction treatment is currently under development for the treatment of whole blood using riboflavin and a higher dose of UV light. We sought to determine the effectiveness of this treatment in prevention of alloimmunization. STUDY DESIGN AND METHODS BALB/c mice were transfused with untreated or UV+R treated allogeneic C57Bl/6 whole blood with or without leukoreduction. Mice were evaluated for donor specific antibodies and ex vivo splenocyte cytokine responses, as well as for changes in the frequency of regulatory T (Treg) cells. RESULTS UV+R treatment blocked cytokine priming and reduced anti-MHC alloantibody responses to transfused whole blood. Leukoreduction reduced alloantibody levels in both the untreated and UV+R groups. Mice transfused with UV+R treated whole blood had reduced alloantibody and cytokine responses when subsequently transfused with untreated blood from the same donor type. This reduction in responses was not associated with increased Treg cells. CONCLUSIONS Pathogen reduction of whole blood with UV+R significantly reduces, but does not eliminate the alloimmune response. Exposure to UV+R treated whole blood transfusion does appear to induce tolerance to alloantigens resulting in reduced anti-MHC alloantibody and cytokine responses to subsequent exposures to the same alloantigens. This tolerance does not appear to be driven by an increase in Treg cells. PMID:27859333

Centrifuge separation effect on bacterial indicator reduction in dairy manure.

PubMed

Liu, Zong; Carroll, Zachary S; Long, Sharon C; Roa-Espinosa, Aicardo; Runge, Troy

2017-04-15

Centrifugation is a commonly applied separation method for manure processing on large farms to separate solids and nutrients. Pathogen reduction is also an important consideration for managing manure. Appropriate treatment reduces risks from pathogen exposure when manure is used as soil amendments or the processed liquid stream is recycled to flush the barn. This study investigated the effects of centrifugation and polymer addition on bacterial indicator removal from the liquid fraction of manure slurries. Farm samples were taken from a manure centrifuge processing system. There were negligible changes of quantified pathogen indicator concentrations in the low-solids centrate compared to the influent slurry. To study if possible improvements could be made to the system, lab scale experiments were performed investigating a range of g-forces and flocculating polymer addition. The results demonstrated that polymer addition had a negligible effect on the indicator bacteria levels when centrifuged at high g forces. However, the higher g force centrifugation was capable of reducing bacterial indicator levels up to two-log 10 in the liquid stream of the manure, although at speeds higher than typical centrifuge operations currently used for manure processing applications. This study suggests manure centrifuge equipment could be redesigned to provide pathogen reduction to meet emerging issues, such as zoonotic pathogen control. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

Evaluation of radio-frequency heating in controlling Salmonella enterica in raw shelled almonds.

PubMed

Jeong, Seul-Gi; Baik, Oon-Doo; Kang, Dong-Hyun

2017-08-02

This study was conducted to investigate the efficacy of radio-frequency (RF) heating to reduce Salmonella enterica serovars Enteritidis, Typhimurium, and Senftenberg in raw shelled almonds compared to conventional convective heating, and the effect of RF heating on quality by measuring changes in the color and degree of lipid oxidation. Agar-grown cells of three pathogens were inoculated onto the surface or inside of raw shelled almonds using surface inoculation or the vacuum perfusion method, respectively, and subjected to RF or conventional heating. RF heating for 40s achieved 3.7-, 6.0-, and 5.6-log reductions in surface-inoculated S. Enteritidis, S. Typhimurium, and S. Senftenberg, respectively, whereas the reduction of these pathogens following convective heating for 600s was 1.7, 2.5, and 3.7 log, respectively. RF heating reduced internally inoculated pathogens to below the detection limit (0.7 logCFU/g) after 30s. However, conventional convective heating did not attain comparable reductions even at the end of treatment (600s). Color values, peroxide values, and acid values of RF-treated (40-s treatment) almonds were not significantly (P>0.05) different from those of nontreated samples. These results suggest that RF heating can be applied to control internalized pathogens as well as surface-adhering pathogens in raw almonds without affecting product quality. Copyright © 2017. Published by Elsevier B.V.

Risk-Based Treatment Targets for Onsite Non-Potable Water ...

EPA Pesticide Factsheets

This presentation presents risk-based enteric pathogen log reduction targets for non-potable and potable uses of a variety of alternative source waters (i.e., municipal wastewater, locally-collected greywater, rainwater, and stormwater). A probabilistic, forward Quantitative Microbial Risk Assessment (QMRA) was used to derive the pathogen log10 reduction targets (LRTs) that corresponded with an infection risk of either 10-4 per person per year (ppy) or 10-2 ppy. The QMRA accounted for variation in pathogen concentration and sporadic pathogen occurrence (when data were available) in source waters for reference pathogens Rotavirus, Adenovirus, Norovirus, Campylobacter spp., Salmonella spp., Giardia spp., and Cryptosporidium spp.. Non-potable uses included indoor use (for toilet flushing and clothes washing) with accidental ingestion of treated non-potable water (or cross connection with potable water), and unrestricted irrigation for outdoor use. Various exposure scenarios captured the uncertainty from key inputs, i.e., the pathogen concentration in source water; the volume of water ingested; and for the indoor use, the frequency of and the fraction of the population exposed to accidental ingestion. Both potable and non-potable uses required pathogen treatment for the selected waters and the LRT was generally greater for potable use than nonpotable indoor use and unrestricted irrigation. The difference in treatment requirements among source waters was driven by th

Comparative analysis of solar pasteurization versus solar disinfection for the treatment of harvested rainwater.

PubMed

Strauss, André; Dobrowsky, Penelope Heather; Ndlovu, Thando; Reyneke, Brandon; Khan, Wesaal

2016-12-09

Numerous pathogens and opportunistic pathogens have been detected in harvested rainwater. Developing countries, in particular, require time- and cost-effective treatment strategies to improve the quality of this water source. The primary aim of the current study was thus to compare solar pasteurization (SOPAS; 70 to 79 °C; 80 to 89 °C; and ≥90 °C) to solar disinfection (SODIS; 6 and 8 hrs) for their efficiency in reducing the level of microbial contamination in harvested rainwater. The chemical quality (anions and cations) of the SOPAS and SODIS treated and untreated rainwater samples were also monitored. While the anion concentrations in all the samples were within drinking water guidelines, the concentrations of lead (Pb) and nickel (Ni) exceeded the guidelines in all the SOPAS samples. Additionally, the iron (Fe) concentrations in both the SODIS 6 and 8 hr samples were above the drinking water guidelines. A >99% reduction in Escherichia coli and heterotrophic bacteria counts was then obtained in the SOPAS and SODIS samples. Ethidium monoazide bromide quantitative polymerase chain reaction (EMA-qPCR) analysis revealed a 94.70% reduction in viable Legionella copy numbers in the SOPAS samples, while SODIS after 6 and 8 hrs yielded a 50.60% and 75.22% decrease, respectively. Similarly, a 99.61% reduction in viable Pseudomonas copy numbers was observed after SOPAS treatment, while SODIS after 6 and 8 hrs yielded a 47.27% and 58.31% decrease, respectively. While both the SOPAS and SODIS systems reduced the indicator counts to below the detection limit, EMA-qPCR analysis indicated that SOPAS treatment yielded a 2- and 3-log reduction in viable Legionella and Pseudomonas copy numbers, respectively. Additionally, SODIS after 8 hrs yielded a 2-log and 1-log reduction in Legionella and Pseudomonas copy numbers, respectively and could be considered as an alternative, cost-effective treatment method for harvested rainwater.

Interventions for the control of diarrhoeal diseases among young children: improving water supplies and excreta disposal facilities*

PubMed Central

Esrey, S. A.; Feachem, R. G.; Hughes, J. M.

1985-01-01

A theoretical model is proposed that relates the level of ingestion of diarrhoea-causing pathogens to the frequency of diarrhoea in the community. The implications of this model are that, in poor communities with inadequate water supply and excreta disposal, reducing the level of enteric pathogen ingestion by a given amount will have a greater impact on diarrhoea mortality rates than on morbidity rates, a greater impact on the incidence rate of severe diarrhoea than on that of mild diarrhoea, and a greater impact on diarrhoea caused by pathogens having high infectious doses than on diarrhoea caused by pathogens of a low infectious dose. The impact of water supply and sanitation on diarrhoea, related infections, nutritional status, and mortality is analysed by reviewing 67 studies from 28 countries. The median reductions in diarrhoea morbidity rates are 22% from all studies and 27% from a few better-designed studies. All studies of the impact on total mortality rates show a median reduction of 21%, while the few better-designed studies give a median reduction of 30%. Improvements in water quality have less of an impact than improvements in water availability or excreta disposal. PMID:3878742

Endogenous System Microbes as Treatment Process ...

EPA Pesticide Factsheets

Monitoring the efficacy of treatment strategies to remove pathogens in decentralized systems remains a challenge. Evaluating log reduction targets by measuring pathogen levels is hampered by their sporadic and low occurrence rates. Fecal indicator bacteria are used in centralized systems to indicate the presence of fecal pathogens, but are ineffective decentralized treatment process indicators as they generally occur at levels too low to assess log reduction targets. System challenge testing by spiking with high loads of fecal indicator organisms, like MS2 coliphage, has limitations, especially for large systems. Microbes that are endogenous to the decentralized system, occur in high abundances and mimic removal rates of bacterial, viral and/or parasitic protozoan pathogens during treatment could serve as alternative treatment process indicators to verify log reduction targets. To identify abundant microbes in wastewater, the bacterial and viral communities were examined using deep sequencing. Building infrastructure-associated bacteria, like Zoogloea, were observed as dominant members of the bacterial community in graywater. In blackwater, bacteriophage of the order Caudovirales constituted the majority of contiguous sequences from the viral community. This study identifies candidate treatment process indicators in decentralized systems that could be used to verify log removal during treatment. The association of the presence of treatment process indic

Tinidazole inhibitory and cidal activity against anaerobic periodontal pathogens.

PubMed

Alou, L; Giménez, M J; Manso, F; Sevillano, D; Torrico, M; González, N; Granizo, J J; Bascones, A; Prieto, J; Maestre, J R; Aguilar, L

2009-05-01

The in vitro activity of tinidazole against anaerobic periodontal pathogens (25 Prevotella buccae, 18 Prevotella denticola, 10 Prevotella intermedia, 6 Prevotella melaninogenica, 5 Prevotella oralis, 10 Fusobacterium nucleatum and 8 Veillonella spp.) was determined by agar dilution. MIC(90) values (minimum inhibitory concentration for 90% of the organisms) were 8 microg/mL for Veillonella spp., 4 microg/mL for P. intermedia, 2 microg/mL for P. buccae, 1 microg/mL for Fusobacterium spp. and 0.5 microg/mL for other Prevotella spp. Cidal activity was studied by killing curves with tinidazole and amoxicillin (alone and in combination) at concentrations similar to those achieved in crevicular fluid (41.2 microg/mL tinidazole and 14.05 microg/mL amoxicillin) against an inoculum of ca. 10(7)colony-forming units/mL of four bacterial groups, each one composed of four different strains of the following periodontal isolates: Prevotella spp., Fusobacterium spp. and Veillonella spp. (anaerobes) and one amoxicillin-susceptible Streptococcus spp. (facultative) in a proportion of 1:1:1:1. When only beta-lactamase-negative Prevotella or Fusobacterium strains were tested, significantly higher reductions were found with amoxicillin (>4 log reduction at 48 h) versus controls. The presence of beta-lactamase-positive Prevotella spp. or F. nucleatum strains rendered amoxicillin inactive (no reductions at 48 h), with no differences from controls. Amoxicillin+tinidazole produced >3 log reduction at 24h and >4 log reduction at 48 h regardless of the presence or not of beta-lactamase-positive strains. The presence in crevicular fluid of beta-lactamases produced by beta-lactamase-positive periodontal pathogens may have ecological and therapeutic consequences since it may protect beta-lactamase-negative periodontal pathogens from amoxicillin treatment. In vitro, tinidazole offered high antianaerobic activity against beta-lactamase-positive and -negative periodontal pathogens, avoiding amoxicillin inactivation.

Irradiation and additive combinations on the pathogen reduction and quality of poultry meat.

PubMed

Ahn, Dong U; Kim, Il Suk; Lee, Eun Joo

2013-02-01

Reduction of foodborne illnesses and deaths by improving the safety of poultry products is one of the priority areas in the United States, and developing and implementing effective food processing technologies can be very effective to accomplish that goal. Irradiation is an effective processing technology for eliminating pathogens in poultry meat. Addition of antimicrobial agents during processing can be another approach to control pathogens in poultry products. However, the adoption of irradiation technology by the meat industry is limited because of quality and health concerns about irradiated meat products. Irradiation produces a characteristic aroma as well as alters meat flavor and color that significantly affect consumer acceptance. The generation of a pink color in cooked poultry and off-odor in poultry by irradiation is a critical issue because consumers associate the presence of a pink color in cooked poultry breast meat as contaminated or undercooked, and off-odor in raw meat and off-flavor in cooked meat with undesirable chemical reactions. As a result, the meat industry has difficulties in using irradiation to achieve its food safety benefits. Antimicrobials such as sodium lactate, sodium diacetate, and potassium benzoate are extensively used to extend the shelf-life and ensure the safety of meat products. However, the use of these antimicrobial agents alone cannot guarantee the safety of poultry products. It is known that some of the herbs, spices, and antimicrobials commonly used in meat processing can have synergistic effects with irradiation in controlling pathogens in meat. Also, the addition of spices or herbs in irradiated meat improves the quality of irradiated poultry by reducing lipid oxidation and production of off-odor volatiles or masking off-flavor. Therefore, combinations of irradiation with these additives can accomplish better pathogen reduction in meat products than using them alone even at lower levels of antimicrobials/herbs and irradiation doses. Effects of irradiation and additive combinations on the pathogen reduction and quality of poultry meat will be discussed in detail.

Health Technology Assessment of pathogen reduction technologies applied to plasma for clinical use

PubMed Central

Cicchetti, Americo; Berrino, Alexandra; Casini, Marina; Codella, Paola; Facco, Giuseppina; Fiore, Alessandra; Marano, Giuseppe; Marchetti, Marco; Midolo, Emanuela; Minacori, Roberta; Refolo, Pietro; Romano, Federica; Ruggeri, Matteo; Sacchini, Dario; Spagnolo, Antonio G.; Urbina, Irene; Vaglio, Stefania; Grazzini, Giuliano; Liumbruno, Giancarlo M.

2016-01-01

Although existing clinical evidence shows that the transfusion of blood components is becoming increasingly safe, the risk of transmission of known and unknown pathogens, new pathogens or re-emerging pathogens still persists. Pathogen reduction technologies may offer a new approach to increase blood safety. The study is the output of collaboration between the Italian National Blood Centre and the Post-Graduate School of Health Economics and Management, Catholic University of the Sacred Heart, Rome, Italy. A large, multidisciplinary team was created and divided into six groups, each of which addressed one or more HTA domains. Plasma treated with amotosalen + UV light, riboflavin + UV light, methylene blue or a solvent/detergent process was compared to fresh-frozen plasma with regards to current use, technical features, effectiveness, safety, economic and organisational impact, and ethical, social and legal implications. The available evidence is not sufficient to state which of the techniques compared is superior in terms of efficacy, safety and cost-effectiveness. Evidence on efficacy is only available for the solvent/detergent method, which proved to be non-inferior to untreated fresh-frozen plasma in the treatment of a wide range of congenital and acquired bleeding disorders. With regards to safety, the solvent/detergent technique apparently has the most favourable risk-benefit profile. Further research is needed to provide a comprehensive overview of the cost-effectiveness profile of the different pathogen-reduction techniques. The wide heterogeneity of results and the lack of comparative evidence are reasons why more comparative studies need to be performed. PMID:27403740

40 CFR 503.30 - Scope.

Code of Federal Regulations, 2011 CFR

2011-07-01

... DISPOSAL OF SEWAGE SLUDGE Pathogens and Vector Attraction Reduction § 503.30 Scope. (a) This subpart... land, forest, or a reclamation site. (d) This subpart contains alternative vector attraction reduction...

40 CFR 503.30 - Scope.

Code of Federal Regulations, 2010 CFR

2010-07-01

... DISPOSAL OF SEWAGE SLUDGE Pathogens and Vector Attraction Reduction § 503.30 Scope. (a) This subpart... land, forest, or a reclamation site. (d) This subpart contains alternative vector attraction reduction...

Essential oils against foodborne pathogens and spoilage bacteria in minced meat.

PubMed

Barbosa, Lidiane Nunes; Rall, Vera Lucia Mores; Fernandes, Ana Angélica Henrique; Ushimaru, Priscila Ikeda; da Silva Probst, Isabella; Fernandes, Ary

2009-01-01

The antimicrobial activity of essential oils of oregano, thyme, basil, marjoram, lemongrass, ginger, and clove was investigated in vitro by agar dilution method and minimal inhibitory concentration (MIC) determination against Gram-positive (Staphylococcus aureus and Listeria monocytogenes) and Gram-negative strains (Escherichia coli and Salmonella Enteritidis). MIC(90%) values were tested against bacterial strains inoculated experimentally in irradiated minced meat and against natural microbiota (aerobic or facultative, mesophilic, and psychrotrophic bacteria) found in minced meat samples. MIC(90%) values ranged from 0.05%v/v (lemongrass oil) to 0.46%v/v (marjoram oil) to Gram-positive bacteria and from 0.10%v/v (clove oil) to 0.56%v/v (ginger oil) to Gram-negative strains. However, the MIC(90%) assessed on minced meat inoculated experimentally with foodborne pathogen strains and against natural microbiota of meat did not show the same effectiveness, and 1.3 and 1.0 were the highest log CFU/g reduction values obtained against tested microorganisms.

Essential Oils Against Foodborne Pathogens and Spoilage Bacteria in Minced Meat

PubMed Central

Barbosa, Lidiane Nunes; Rall, Vera Lucia Mores; Fernandes, Ana Angélica Henrique; Ushimaru, Priscila Ikeda; da Silva Probst, Isabella

2009-01-01

Abstract The antimicrobial activity of essential oils of oregano, thyme, basil, marjoram, lemongrass, ginger, and clove was investigated in vitro by agar dilution method and minimal inhibitory concentration (MIC) determination against Gram-positive (Staphylococcus aureus and Listeria monocytogenes) and Gram-negative strains (Escherichia coli and Salmonella Enteritidis). MIC90% values were tested against bacterial strains inoculated experimentally in irradiated minced meat and against natural microbiota (aerobic or facultative, mesophilic, and psychrotrophic bacteria) found in minced meat samples. MIC90% values ranged from 0.05%v/v (lemongrass oil) to 0.46%v/v (marjoram oil) to Gram-positive bacteria and from 0.10%v/v (clove oil) to 0.56%v/v (ginger oil) to Gram-negative strains. However, the MIC90% assessed on minced meat inoculated experimentally with foodborne pathogen strains and against natural microbiota of meat did not show the same effectiveness, and 1.3 and 1.0 were the highest log CFU/g reduction values obtained against tested microorganisms. PMID:19580445

Efficacy of microorganisms selected from compost to control soil-borne pathogens.

PubMed

Pugliese, M; Gullino, M L; Garibaldi, A

2010-01-01

Suppression of soil-borne plant pathogens with compost has been widely studied. Compost has been found to be suppressive against several soil-borne pathogens in various cropping systems. However, an increase of some diseases due to compost usage has also been observed, since compost is a product that varies considerably in chemical, physical and biotic composition, and, consequently, also in ability to suppress soil borne diseases. New opportunities in disease management can be obtained by the selection of antagonists from suppressive composts. The objective of the present work was to isolate microorganisms from a suppressive compost and to test them for their activity against soil-borne pathogens. A compost from green wastes, organic domestic wastes and urban sludge's that showed a good suppressive activity in previous trials was used as source of microorganisms. Serial diluted suspensions of compost samples were plated on five different media: selective for Fusarium sp., selective for Trichoderma sp., selective for oomycetes, potato dextrose agar (PDA) for isolation of fungi, lysogeny broth (LB) for isolation of bacteria. In total, 101 colonies were isolated from plates and tested under laboratory conditions on tomato seedlings growing on perlite medium in Petri plates infected with Fusarium oxysporum f.sp. radicis-lycopersici and compared to a commercial antagonist (Streptomyces griserovidis, Mycostop, Bioplanet). Among them, 28 showed a significant disease reduction and were assessed under greenhouse condition on three pathosystems: Fusarium oxysporum f.sp. basilica/basil, Phytophthora nicotianae/tomato and Rhizoctonia solani/bean. Fusarium spp. selected from compost generally showed a good disease control against Fusarium wilts, while only bacteria significantly controlled P. nicotianae on tomato under greenhouse conditions. None of the microorganisms was able to control the three soil-borne pathogens together, in particular Rhizoctonia solani. Results confirmed the good suppressive activity of the compost under study against soil-borne pathogens. The selection of antagonists from compost is a promising strategy for the development of new biological control agents against soil-borne pathogens.

Alternative Treatment Technologies – Working With the Pathogen Equivalency Committee

EPA Science Inventory

Under current Federal regulations (40 CFR 503), municipal sludge must be treated prior to land application. The regulations identify two classes of treatment with respect to pathogen reduction: Class B (three alternatives) which provides a minimum acceptable level of treatment;...

Hydrothermal carbonization of livestock mortality for the reduction of pathogens and microbially-derived DNA

USDA-ARS?s Scientific Manuscript database

Hydrothermal carbonization (HTC), utilizing high temperature and pressure, has the potential to treat agricultural waste and inactivate pathogens, antibiotic resistance genes (ARG), and contaminants of emerging concern (CEC) in an environmentally and economically friendly manner. Livestock mortality...

Evaluation of near-infrared pasteurization in controlling Escherichia coli O157:H7, Salmonella enterica serovar typhimurium, and Listeria monocytogenes in ready-to-eat sliced ham.

PubMed

Ha, Jae-Won; Ryu, Sang-Ryeol; Kang, Dong-Hyun

2012-09-01

This study was conducted to investigate the efficacy of near-infrared (NIR) heating to reduce Salmonella enterica serovar Typhimurium, Escherichia coli O157:H7, and Listeria monocytogenes in ready-to-eat (RTE) sliced ham compared to conventional convective heating, and the effect of NIR heating on quality was determined by measuring the color and texture change. A cocktail of three pathogens was inoculated on the exposed or protected surfaces of ham slices, followed by NIR or conventional heating at 1.8 kW. NIR heating for 50 s achieved 4.1-, 4.19-, and 3.38-log reductions in surface-inoculated S. Typhimurium, E. coli O157:H7, and L. monocytogenes, respectively, whereas convective heating needed 180 s to attain comparable reductions for each pathogen. There were no statistically significant (P > 0.05) differences in reduction between surface- and internally inoculated pathogens at the end of NIR treatment (50 s). However, when treated with conventional convective heating, significant (P < 0.05) differences were observed at the final stages of the treatment (150 and 180 s). Color values and texture parameters of NIR-treated (50-s treatment) ham slices were not significantly (P > 0.05) different from those of nontreated samples. These results suggest that NIR heating can be applied to control internalized pathogens as well as surface-adhering pathogens in RTE sliced meats without affecting product quality.

Evaluation of Near-Infrared Pasteurization in Controlling Escherichia coli O157:H7, Salmonella enterica Serovar Typhimurium, and Listeria monocytogenes in Ready-To-Eat Sliced Ham

PubMed Central

Ha, Jae-Won; Ryu, Sang-Ryeol

2012-01-01

This study was conducted to investigate the efficacy of near-infrared (NIR) heating to reduce Salmonella enterica serovar Typhimurium, Escherichia coli O157:H7, and Listeria monocytogenes in ready-to-eat (RTE) sliced ham compared to conventional convective heating, and the effect of NIR heating on quality was determined by measuring the color and texture change. A cocktail of three pathogens was inoculated on the exposed or protected surfaces of ham slices, followed by NIR or conventional heating at 1.8 kW. NIR heating for 50 s achieved 4.1-, 4.19-, and 3.38-log reductions in surface-inoculated S. Typhimurium, E. coli O157:H7, and L. monocytogenes, respectively, whereas convective heating needed 180 s to attain comparable reductions for each pathogen. There were no statistically significant (P > 0.05) differences in reduction between surface- and internally inoculated pathogens at the end of NIR treatment (50 s). However, when treated with conventional convective heating, significant (P < 0.05) differences were observed at the final stages of the treatment (150 and 180 s). Color values and texture parameters of NIR-treated (50-s treatment) ham slices were not significantly (P > 0.05) different from those of nontreated samples. These results suggest that NIR heating can be applied to control internalized pathogens as well as surface-adhering pathogens in RTE sliced meats without affecting product quality. PMID:22773635

Inactivation of Staphylococcus aureus and native microflora in human milk by high pressure processing

NASA Astrophysics Data System (ADS)

Windyga, Bożena; Rutkowska, Małgorzata; Sokołowska, Barbara; Skąpska, Sylwia; Wesołowska, Aleksandra; Wilińska, Maria; Fonberg-Broczek, Monika; Rzoska, Sylwester J.

2015-04-01

The storage of unpreserved food, including breast milk, is associated with the growth of microorganisms, including pathogenic bacteria. It is therefore necessary to use suitable processes to eliminate pathogenic microorganisms and reduce the total microbial count in order to ensure product safety for consumers. In the present study, samples of milk obtained from volunteers donating to the human milk bank were artificially contaminated with Staphylococcus aureus ATCC 6538. This bacteria was the model microorganism of choice, being relatively resistant to high pressure as well as posing the most serious risk to infant health. The results obtained show that high pressure processing can reduce the count of S. aureus by about 5 log units at 4°C and about 8 log units at 50°C, and totally eliminate Enterobacteriaceae after 5 min of treatment, and result in a total microbial count reduction after 10 min treatment at 500 MPa at 20°C and 50°C. This suggests the possibility of this technology being applied to ensure the adequate safety and quality of human breast milk in human milk banks. This paper was presented at the LIIth European High Pressure Research Group (EHPRG 52) Meeting in Lyon (France), 7-12 September 2014.

Pathogen reduction requirements for direct potable reuse in Antarctica: evaluating human health risks in small communities.

PubMed

Barker, S Fiona; Packer, Michael; Scales, Peter J; Gray, Stephen; Snape, Ian; Hamilton, Andrew J

2013-09-01

Small, remote communities often have limited access to energy and water. Direct potable reuse of treated wastewater has recently gained attention as a potential solution for water-stressed regions, but requires further evaluation specific to small communities. The required pathogen reduction needed for safe implementation of direct potable reuse of treated sewage is an important consideration but these are typically quantified for larger communities and cities. A quantitative microbial risk assessment (QMRA) was conducted, using norovirus, giardia and Campylobacter as reference pathogens, to determine the level of treatment required to meet the tolerable annual disease burden of 10(-6) DALYs per person per year, using Davis Station in Antarctica as an example of a small remote community. Two scenarios were compared: published municipal sewage pathogen loads and estimated pathogen loads during a gastroenteritis outbreak. For the municipal sewage scenario, estimated required log10 reductions were 6.9, 8.0 and 7.4 for norovirus, giardia and Campylobacter respectively, while for the outbreak scenario the values were 12.1, 10.4 and 12.3 (95th percentiles). Pathogen concentrations are higher under outbreak conditions as a function of the relatively greater degree of contact between community members in a small population, compared with interactions in a large city, resulting in a higher proportion of the population being at risk of infection and illness. While the estimates of outbreak conditions may overestimate sewage concentration to some degree, the results suggest that additional treatment barriers would be required to achieve regulatory compliance for safe drinking water in small communities. Copyright © 2013 Elsevier B.V. All rights reserved.

Evidence of immunocompetence reduction induced by cadmium exposure in honey bees (Apis mellifera).

PubMed

Polykretis, P; Delfino, G; Petrocelli, I; Cervo, R; Tanteri, G; Montori, G; Perito, B; Branca, J J V; Morucci, G; Gulisano, M

2016-11-01

In the last decades a dramatic loss of Apis mellifera hives has been reported in both Europe and USA. Research in this field is oriented towards identifying a synergy of contributing factors, i.e. pathogens, pesticides, habitat loss and pollution to the weakening of the hive. Cadmium (Cd) is a hazardous anthropogenic pollutant whose effects are proving to be increasingly lethal. Among the multiple damages related to Cd contamination, some studies report that it causes immunosuppression in various animal species. The aim of this paper is to determine whether contamination by Cd, may have a similar effect on the honey bees' immunocompetence. Our results, obtained by immune challenge experiments and confirmed by structural and ultrastructural observations show that such metal causes a reduction in immunocompetence in 3 days Cd exposed bees. As further evidence of honey bee response to Cd treatment, Energy Dispersive X-ray Spectroscopy (X-EDS) has revealed the presence of zinc (Zn) in peculiar electron-dense granules in fat body cells. Zn is a characteristic component of metallothioneins (MTs), which are usually synthesized as anti-oxidant and scavenger tools against Cd contamination. Our findings suggest that honey bee colonies may have a weakened immune system in Cd polluted areas, resulting in a decreased ability in dealing with pathogens. Copyright © 2016 Elsevier Ltd. All rights reserved.

Co-infections determine patterns of mortality in a population exposed to parasite infection.

PubMed

Woolhouse, Mark E J; Thumbi, Samuel M; Jennings, Amy; Chase-Topping, Margo; Callaby, Rebecca; Kiara, Henry; Oosthuizen, Marinda C; Mbole-Kariuki, Mary N; Conradie, Ilana; Handel, Ian G; Poole, E Jane; Njiiri, Evalyne; Collins, Nicola E; Murray, Gemma; Tapio, Miika; Auguet, Olga Tosas; Weir, Willie; Morrison, W Ivan; Kruuk, Loeske E B; Bronsvoort, B Mark de C; Hanotte, Olivier; Coetzer, Koos; Toye, Philip G

2015-03-01

Many individual hosts are infected with multiple parasite species, and this may increase or decrease the pathogenicity of the infections. This phenomenon is termed heterologous reactivity and is potentially an important determinant of both patterns of morbidity and mortality and of the impact of disease control measures at the population level. Using infections with Theileria parva (a tick-borne protozoan, related to Plasmodium) in indigenous African cattle [where it causes East Coast fever (ECF)] as a model system, we obtain the first quantitative estimate of the effects of heterologous reactivity for any parasitic disease. In individual calves, concurrent co-infection with less pathogenic species of Theileria resulted in an 89% reduction in mortality associated with T. parva infection. Across our study population, this corresponds to a net reduction in mortality due to ECF of greater than 40%. Using a mathematical model, we demonstrate that this degree of heterologous protection provides a unifying explanation for apparently disparate epidemiological patterns: variable disease-induced mortality rates, age-mortality profiles, weak correlations between the incidence of infection and disease (known as endemic stability), and poor efficacy of interventions that reduce exposure to multiple parasite species. These findings can be generalized to many other infectious diseases, including human malaria, and illustrate how co-infections can play a key role in determining population-level patterns of morbidity and mortality due to parasite infections.

Endogenous System Microbes as Treatment Process Indicators for Decentralized Non-potable Water Reuse

EPA Science Inventory

Monitoring the efficacy of treatment strategies to remove pathogens in decentralized systems remains a challenge. Evaluating log reduction targets by measuring pathogen levels is hampered by their sporadic and low occurrence rates. Fecal indicator bacteria are used in centraliz...

Inhibition of Escherichia coli O157:H7 and Salmonella enterica on spinach and identification of antimicrobial substances produced by a commercial Lactic Acid Bacteria food safety intervention.

PubMed

Cálix-Lara, Thelma F; Rajendran, Mahitha; Talcott, Stephen T; Smith, Stephen B; Miller, Rhonda K; Castillo, Alejandro; Sturino, Joseph M; Taylor, T Matthew

2014-04-01

The microbiological safety of fresh produce is of concern for the U.S. food supply. Members of the Lactic Acid Bacteria (LAB) have been reported to antagonize pathogens by competing for nutrients and by secretion of substances with antimicrobial activity, including organic acids, peroxides, and antimicrobial polypeptides. The objectives of this research were to: (i) determine the capacity of a commercial LAB food antimicrobial to inhibit Escherichia coli O157:H7 and Salmonella enterica on spinach leaf surfaces, and (ii) identify antimicrobial substances produced in vitro by the LAB comprising the food antimicrobial. Pathogens were inoculated on freshly harvested spinach, followed by application of the LAB antimicrobial. Treated spinach was aerobically incubated up to 12 days at 7 °C and surviving pathogens enumerated via selective/differential plating. l-Lactic acid and a bacteriocin-like inhibitory substance (BLIS) were detected and quantified from cell-free fermentates obtained from LAB-inoculated liquid microbiological medium. Application of 8.0 log10 CFU/g LAB produced significant (p < 0.05) reductions in E. coli O157:H7 and Salmonella populations on spinach of 1.6 and 1.9 log10 CFU/g, respectively. It was concluded the LAB antimicrobial inhibited foodborne pathogens on spinach during refrigerated storage, likely the result of the production of metabolites with antimicrobial activity. Copyright © 2013 Elsevier Ltd. All rights reserved.

Effect of water coagulation by seeds of Moringa oleifera on bacterial concentrations.

PubMed

Madsen, M; Schlundt, J; Omer, E F

1987-06-01

The effects of a Sudanese water purification method traditionally used in Sudan to treat turbid waters were studied with respect to turbidity reduction and removal of faecal indicator bacteria as well as selected enteric bacterial pathogens. Water treatment was performed at 30 degrees C with Moringa oleifera seed material as a coagulant, and the technique employed corresponded closely to that used to clarify turbid water in Sudanese villages. A turbidity reduction of 80.0-99.5% paralleled by a primary bacterial reduction of 1-4 log units (90.00-99.99%) was obtained within the first 1 to 2 h of treatment, the bacteria being concentrated in the coagulated sediment. During the 24 h observation period a secondary bacterial increase due to regrowth in the supernatant water was consistently observed for Salmonella typhimurium and Shigella sonnei, in some cases for Escherichia coli, but not for Vibrio cholerae, Streptococcus faecalis and Clostridium perfringens. The potential of the method when compared with some alternative for the improvement of rural drinking water supplies is discussed.

Quo vadis? – Monitoring Campylobacter in Germany

PubMed Central

Stingl, K.; Knüver, M.-T.; Vogt, P.; Buhler, C.; Krüger, N.-J.; Alt, K.; Tenhagen, B.-A.; Hartung, M.; Schroeter, A.; Ellerbroek, L.; Appel, B.; Käsbohrer, A.

2012-01-01

Campylobacter is a poorly recognized foodborne pathogen, leading the statistics of bacterially caused human diarrhoea in Europe during the last years. In this review, we present qualitative and quantitative German data obtained in the framework of specific monitoring programs and from routine surveillance. These also comprise recent data on antimicrobial resistances of food isolates. Due to the considerable reduction of in vitro growth capabilities of stressed bacteria, there is a clear discrepancy between the detection limit of Campylobacter by cultivation and its infection potential. Moreover, antimicrobial resistances of Campylobacter isolates established during fattening of livestock are alarming, since they constitute an additional threat to human health. The European Food Safety Authority (EFSA) discusses the establishment of a quantitative limit for Campylobacter contamination of broiler carcasses in order to achieve an appropriate level of protection for consumers. Currently, a considerable amount of German broiler carcasses would not comply with this future criterion. We recommend Campylobacter reduction strategies to be focussed on the prevention of fecal contamination during slaughter. Decontamination is only a sparse option, since the reduction efficiency is low and its success depends on the initial contamination concentration. PMID:24611125

Simulation of enteric pathogen concentrations in locally-collected greywater and wastewater for microbial risk assessments

EPA Science Inventory

As decentralized water reuse continues to gain popularity, risk-based treatment guidance is increasingly sought for the protection of public health. However, efforts to evaluate pathogen risks and log-reduction requirements have been hindered by an incomplete understanding of pat...

Pathogen reduction through additive-free short-wave UV light irradiation retains the optimal efficacy of human platelet lysate for the expansion of human bone marrow mesenchymal stem cells.

PubMed

Viau, Sabrina; Chabrand, Lucie; Eap, Sandy; Lorant, Judith; Rouger, Karl; Goudaliez, Francis; Sumian, Chryslain; Delorme, Bruno

2017-01-01

We recently developed and characterized a standardized and clinical grade human Platelet Lysate (hPL) that constitutes an advantageous substitute for fetal bovine serum (FBS) for human mesenchymal stem cell (hMSC) expansion required in cell therapy procedures, avoiding xenogenic risks (virological and immunological) and ethical issues. Because of the progressive use of pathogen-reduced (PR) labile blood components, and the requirement of ensuring the viral safety of raw materials for cell therapy products, we evaluated the impact of the novel procedure known as THERAFLEX UV-Platelets for pathogen reduction on hPL quality (growth factors content) and efficacy (as a medium supplement for hMSC expansion). This technology is based on short-wave ultraviolet light (UV-C) that induces non-reversible damages in DNA and RNA of pathogens while preserving protein structures and functions, and has the main advantage of not needing the addition of any photosensitizing additives (that might secondarily interfere with hMSCs). We applied the THERAFLEX UV-Platelets procedure on fresh platelet concentrates (PCs) suspended in platelet additive solution and prepared hPL from these treated PCs. We compared the quality and efficacy of PR-hPL with the corresponding non-PR ones. We found no impact on the content of five cytokines tested (EGF, bFGF, PDGF-AB, VEGF and IGF-1) but a significant decrease in TGF-ß1 (-21%, n = 11, p<0.01). We performed large-scale culture of hMSCs from bone marrow (BM) during three passages and showed that hPL or PR-hPL at 8% triggered comparable BM-hMSC proliferation as FBS at 10% plus bFGF. Moreover, after proliferation of hMSCs in an hPL- or PR-hPL-containing medium, their profile of membrane marker expression, their clonogenic potential and immunosuppressive properties were maintained, in comparison with BM-hMSCs cultured under FBS conditions. The potential to differentiate towards the adipogenic and osteogenic lineages of hMSCs cultured in parallel in the three conditions also remained identical. We demonstrated the feasibility of using UV-C-treated platelets to subsequently obtain pathogen-reduced hPL, while preserving its optimal quality and efficacy for hMSC expansion in cell therapy applications.

Survival of Salmonella and Escherichia coli O157:H7 on strawberries, basil, and other leafy greens during storage.

PubMed

Delbeke, Stefanie; Ceuppens, Siele; Jacxsens, Liesbeth; Uyttendaele, Mieke

2015-04-01

The survival of Salmonella and Escherichia coli O157:H7 on strawberries, basil leaves, and other leafy greens (spinach leaves, lamb and butterhead lettuce leaves, baby leaves, and fresh-cut iceberg lettuce) was assessed at cold (<7 °C) and ambient temperatures. All commodities were spot inoculated with E. coli O157:H7 or Salmonella to obtain an initial inoculum of 5 to 6 log and 4 to 5 log CFU/g for strawberries and leafy greens, respectively. Samples were air packed. Strawberries were stored at 4, 10, 15, and 22 °C and basil leaves and other leafy greens at 7, 15, and 22 °C for up to 7 days (or less if spoiled before). Both Salmonella and E. coli O157:H7 showed a gradual decrease in numbers if inoculated on strawberries, with a similar reduction observed at 4, 10, and 15 °C (2 to 3 log after 5 days). However, at 15 °C (and 10 °C for E. coli O157:H7), the survival experiment stopped before day 7, as die-off of both pathogens below the lower limit of detection was achieved or spoilage occurred. At 22 °C, strawberries were moldy after 2 or 4 days. At that time, a 1- to 2-log reduction of both pathogens had occurred. A restricted die-off (on average 1.0 log) and increase (on average , 0.5 log) of both pathogens on basil leaves occurred after 7 days of storage at 7 and 22 °C, respectively. On leafy greens, a comparable decrease as on basil was observed after 3 days at 7 °C. At 22 °C, both pathogens increased to higher numbers on fresh-cut iceberg and butterhead lettuce leaves (on average 1.0 log), probably due to the presence of exudates. However, by using spot inoculation, the increase was rather limited, probably due to minimized contact between the inoculum and cell exudates. Avoiding contamination, in particular, at cultivation (and harvest or postharvest) is important, as both pathogens survive during storage, and strawberries, basil, and other leafy green leaves are consumed without inactivation treatment.

Pathogen reduction through additive-free short-wave UV light irradiation retains the optimal efficacy of human platelet lysate for the expansion of human bone marrow mesenchymal stem cells

PubMed Central

Viau, Sabrina; Chabrand, Lucie; Eap, Sandy; Lorant, Judith; Rouger, Karl; Goudaliez, Francis; Sumian, Chryslain; Delorme, Bruno

2017-01-01

Background We recently developed and characterized a standardized and clinical grade human Platelet Lysate (hPL) that constitutes an advantageous substitute for fetal bovine serum (FBS) for human mesenchymal stem cell (hMSC) expansion required in cell therapy procedures, avoiding xenogenic risks (virological and immunological) and ethical issues. Because of the progressive use of pathogen-reduced (PR) labile blood components, and the requirement of ensuring the viral safety of raw materials for cell therapy products, we evaluated the impact of the novel procedure known as THERAFLEX UV-Platelets for pathogen reduction on hPL quality (growth factors content) and efficacy (as a medium supplement for hMSC expansion). This technology is based on short-wave ultraviolet light (UV-C) that induces non-reversible damages in DNA and RNA of pathogens while preserving protein structures and functions, and has the main advantage of not needing the addition of any photosensitizing additives (that might secondarily interfere with hMSCs). Methodology / Principal findings We applied the THERAFLEX UV-Platelets procedure on fresh platelet concentrates (PCs) suspended in platelet additive solution and prepared hPL from these treated PCs. We compared the quality and efficacy of PR-hPL with the corresponding non-PR ones. We found no impact on the content of five cytokines tested (EGF, bFGF, PDGF-AB, VEGF and IGF-1) but a significant decrease in TGF-ß1 (-21%, n = 11, p<0.01). We performed large-scale culture of hMSCs from bone marrow (BM) during three passages and showed that hPL or PR-hPL at 8% triggered comparable BM-hMSC proliferation as FBS at 10% plus bFGF. Moreover, after proliferation of hMSCs in an hPL- or PR-hPL-containing medium, their profile of membrane marker expression, their clonogenic potential and immunosuppressive properties were maintained, in comparison with BM-hMSCs cultured under FBS conditions. The potential to differentiate towards the adipogenic and osteogenic lineages of hMSCs cultured in parallel in the three conditions also remained identical. Conclusion / Significance We demonstrated the feasibility of using UV-C-treated platelets to subsequently obtain pathogen-reduced hPL, while preserving its optimal quality and efficacy for hMSC expansion in cell therapy applications. PMID:28763452

Bacterial community dynamics in a cooling tower with emphasis on pathogenic bacteria and Legionella species using universal and genus-specific deep sequencing.

PubMed

Pereira, Rui P A; Peplies, Jörg; Höfle, Manfred G; Brettar, Ingrid

2017-10-01

Cooling towers are the major source of outbreaks of legionellosis in Europe and worldwide. These outbreaks are mostly associated with Legionella species, primarily L. pneumophila, and its surveillance in cooling tower environments is of high relevance to public health. In this study, a combined NGS-based approach was used to study the whole bacterial community, specific waterborne and water-based bacterial pathogens, especially Legionella species, targeting the 16S rRNA gene. This approach was applied to water from a cooling tower obtained by monthly sampling during two years. The studied cooling tower was an open circuit cooling tower with lamellar cooling situated in Braunschweig, Germany. A highly diverse bacterial community was observed with 808 genera including 25 potentially pathogenic taxa using universal 16S rRNA primers. Sphingomonas and Legionella were the most abundant pathogenic genera. By applying genus-specific primers for Legionella, a diverse community with 85 phylotypes, and a representative core community with substantial temporal heterogeneity was observed. A high percentage of sequences (65%) could not be affiliated to an acknowledged species. L. pneumophila was part of the core community and the most abundant Legionella species reinforcing the importance of cooling towers as its environmental reservoir. Major temperature shifts (>10 °C) were the key environmental factor triggering the reduction or dominance of the Legionella species in the Legionella community dynamics. In addition, interventions by chlorine dioxide had a strong impact on the Legionella community composition but not on the whole bacterial community. Overall, the presented results demonstrated the value of a combined NGS approach for the molecular monitoring and surveillance of health related pathogens in man-made freshwater systems. Copyright © 2017 Elsevier Ltd. All rights reserved.

Ammonia disinfection of hatchery waste for elimination of single-stranded RNA viruses.

PubMed

Emmoth, Eva; Ottoson, Jakob; Albihn, Ann; Belák, Sándor; Vinnerås, Björn

2011-06-01

Hatchery waste, an animal by-product of the poultry industry, needs sanitation treatment before further use as fertilizer or as a substrate in biogas or composting plants, owing to the potential presence of opportunistic pathogens, including zoonotic viruses. Effective sanitation is also important in viral epizootic outbreaks and as a routine, ensuring high hygiene standards on farms. This study examined the use of ammonia at different concentrations and temperatures to disinfect hatchery waste. Inactivation kinetics of high-pathogenic avian influenza virus H7N1 and low-pathogenic avian influenza virus H5N3, as representatives of notifiable avian viral diseases, were determined in spiked hatchery waste. Bovine parainfluenza virus type 3, feline coronavirus, and feline calicivirus were used as models for other important avian pathogens, such as Newcastle disease virus, infectious bronchitis virus, and avian hepatitis E virus. Bacteriophage MS2 was also monitored as a stable indicator. Coronavirus was the most sensitive virus, with decimal reduction (D) values of 1.2 and 0.63 h after addition of 0.5% (wt/wt) ammonia at 14 and 25°C, respectively. Under similar conditions, high-pathogenic avian influenza H7N1 was the most resistant, with D values of 3.0 and 1.4 h. MS2 was more resistant than the viruses to all treatments and proved to be a suitable indicator of viral inactivation. The results indicate that ammonia treatment of hatchery waste is efficient in inactivating enveloped and naked single-stranded RNA viruses. Based on the D values and confidence intervals obtained, guidelines for treatment were proposed, and one was successfully validated at full scale at a hatchery, with MS2 added to hatchery waste.

Enhanced Inactivation of Food-Borne Pathogens in Ready-To-Eat Sliced Ham by Near-Infrared Heating Combined with UV-C Irradiation and Mechanism of the Synergistic Bactericidal Action

PubMed Central

Ha, Jae-Won

2014-01-01

The objective of the study described in this article was, first, to investigate the effect of the simultaneous application of near-infrared (NIR) heating and UV irradiation on inactivation of Escherichia coli O157:H7, Salmonella enterica serovar Typhimurium, and Listeria monocytogenes in ready-to-eat (RTE) sliced ham and as well as its effect on product quality and, second, to elucidate the underlying mechanisms of the synergistic bactericidal action of NIR heating and UV irradiation. With the inoculation amounts used, simultaneous NIR-UV combined treatment for 70 s achieved 3.62, 4.17, and 3.43 log CFU reductions of E. coli O157:H7, S. Typhimurium, and L. monocytogenes, respectively. For all three pathogens, the simultaneous application of both technologies resulted in an additional log unit reduction as a result of their synergism compared to the sum of the reductions obtained after the individual treatments. To investigate the mechanisms of NIR-UV synergistic injury for a particular microorganism in a food base, we evaluated the effect of four types of metabolic inhibitors using the overlay method and confirmed that damage to cellular membranes and the inability of cells to repair these structures due to ribosomal damage were the primary factors related to the synergistic lethal effect. Additionally, NIR-UV combined treatment for a maximum of 70 s did not alter the color values or texture parameters of ham slices significantly (P > 0.05). These results suggest that a NIR-UV combined process could be an innovative antimicrobial intervention for RTE meat products. PMID:25107964

Enhanced inactivation of food-borne pathogens in ready-to-eat sliced ham by near-infrared heating combined with UV-C irradiation and mechanism of the synergistic bactericidal action.

PubMed

Ha, Jae-Won; Kang, Dong-Hyun

2015-01-01

The objective of the study described in this article was, first, to investigate the effect of the simultaneous application of near-infrared (NIR) heating and UV irradiation on inactivation of Escherichia coli O157:H7, Salmonella enterica serovar Typhimurium, and Listeria monocytogenes in ready-to-eat (RTE) sliced ham and as well as its effect on product quality and, second, to elucidate the underlying mechanisms of the synergistic bactericidal action of NIR heating and UV irradiation. With the inoculation amounts used, simultaneous NIR-UV combined treatment for 70 s achieved 3.62, 4.17, and 3.43 log CFU reductions of E. coli O157:H7, S. Typhimurium, and L. monocytogenes, respectively. For all three pathogens, the simultaneous application of both technologies resulted in an additional log unit reduction as a result of their synergism compared to the sum of the reductions obtained after the individual treatments. To investigate the mechanisms of NIR-UV synergistic injury for a particular microorganism in a food base, we evaluated the effect of four types of metabolic inhibitors using the overlay method and confirmed that damage to cellular membranes and the inability of cells to repair these structures due to ribosomal damage were the primary factors related to the synergistic lethal effect. Additionally, NIR-UV combined treatment for a maximum of 70 s did not alter the color values or texture parameters of ham slices significantly (P > 0.05). These results suggest that a NIR-UV combined process could be an innovative antimicrobial intervention for RTE meat products. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

UPDATE ON PEC ACTIVITIES INCLUDING NEW EVALUATION CRITERIA, THE APPLICATION COMPLETENESS CHECKLIST, AND STATUS OF THE WEBSITE DEVELOPMENT

EPA Science Inventory

US EPA's Pathogen Equivalency Committee (PEC) has updated the evaluation criteria it uses to make recommendations of equivalency (to processes acceptable under 40CFR503) on innovative or alternative sludge pathogen reduction processes. These criteria will be presented along with ...

Physical and chemical interventions to mitigate risk associated with leafy greens

USDA-ARS?s Scientific Manuscript database

Contamination of leafy green vegetables with human pathogens is a source of ongoing concern for consumers. Conventional treatments have typically been able to achieve 1-2 logs reductions of such pathogens as Salmonella, Escherichia coli O157:H7 and Listeria monocytogenes. Novel approaches and treatm...

Enhanced salmonella reduction on tomatoes washed in chlorinated water with wash aid T-128

USDA-ARS?s Scientific Manuscript database

Chlorine is widely used by the fresh and fresh-cut produce industries to reduce microbial populations and to prevent potential pathogen cross contamination during produce washing. However, the organic materials released from produce quickly react with chlorine and degrade its efficacy for pathogen i...

Review of pathogen treatment reductions for onsite non-potable reuse of alternative source waters

EPA Science Inventory

Communities face a challenge when implementing onsite reuse of collected waters for non-potable purposes given the lack of national microbial standards. Quantitative Microbial Risk Assessment (QMRA) can be used to predict the pathogen risks associated with the non-potable reuse o...

The Effect of Antibiotic Exposure and Specimen Volume on the Detection of Bacterial Pathogens in Children With Pneumonia.

PubMed

Driscoll, Amanda J; Deloria Knoll, Maria; Hammitt, Laura L; Baggett, Henry C; Brooks, W Abdullah; Feikin, Daniel R; Kotloff, Karen L; Levine, Orin S; Madhi, Shabir A; O'Brien, Katherine L; Scott, J Anthony G; Thea, Donald M; Howie, Stephen R C; Adrian, Peter V; Ahmed, Dilruba; DeLuca, Andrea N; Ebruke, Bernard E; Gitahi, Caroline; Higdon, Melissa M; Kaewpan, Anek; Karani, Angela; Karron, Ruth A; Mazumder, Razib; McLellan, Jessica; Moore, David P; Mwananyanda, Lawrence; Park, Daniel E; Prosperi, Christine; Rhodes, Julia; Saifullah, Md; Seidenberg, Phil; Sow, Samba O; Tamboura, Boubou; Zeger, Scott L; Murdoch, David R

2017-06-15

Antibiotic exposure and specimen volume are known to affect pathogen detection by culture. Here we assess their effects on bacterial pathogen detection by both culture and polymerase chain reaction (PCR) in children. PERCH (Pneumonia Etiology Research for Child Health) is a case-control study of pneumonia in children aged 1-59 months investigating pathogens in blood, nasopharyngeal/oropharyngeal (NP/OP) swabs, and induced sputum by culture and PCR. Antibiotic exposure was ascertained by serum bioassay, and for cases, by a record of antibiotic treatment prior to specimen collection. Inoculated blood culture bottles were weighed to estimate volume. Antibiotic exposure ranged by specimen type from 43.5% to 81.7% in 4223 cases and was detected in 2.3% of 4863 controls. Antibiotics were associated with a 45% reduction in blood culture yield and approximately 20% reduction in yield from induced sputum culture. Reduction in yield of Streptococcus pneumoniae from NP culture was approximately 30% in cases and approximately 32% in controls. Several bacteria had significant but marginal reductions (by 5%-7%) in detection by PCR in NP/OP swabs from both cases and controls, with the exception of S. pneumoniae in exposed controls, which was detected 25% less frequently compared to nonexposed controls. Bacterial detection in induced sputum by PCR decreased 7% for exposed compared to nonexposed cases. For every additional 1 mL of blood culture specimen collected, microbial yield increased 0.51% (95% confidence interval, 0.47%-0.54%), from 2% when volume was ≤1 mL to approximately 6% for ≥3 mL. Antibiotic exposure and blood culture volume affect detection of bacterial pathogens in children with pneumonia and should be accounted for in studies of etiology and in clinical management. © The Author 2017. Published by Oxford University Press for the Infectious Diseases Society of America.

Combination treatment of chlorine dioxide gas and aerosolized sanitizer for inactivating foodborne pathogens on spinach leaves and tomatoes.

PubMed

Park, Sang-Hyun; Kang, Dong-Hyun

2015-08-17

The objective of this study was to evaluate the antimicrobial effect of chlorine dioxide (ClO2) gas and aerosolized sanitizer, when applied alone or in combination, on the survival of Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes inoculated onto spinach leaves and tomato surfaces. Spinach leaves and tomatoes were inoculated with a cocktail of three strains each of the three foodborne pathogens. ClO2 gas (5 or 10 ppmv) and aerosolized peracetic acid (PAA) (80 ppm) were applied alone or in combination for 20 min. Exposure to 10 ppmv of ClO2 gas for 20 min resulted in 3.4, 3.3, and 3.4 log reductions of E. coli O157:H7, S. Typhimurium, and L. monocytogenes on spinach leaves, respectively. Treatment with 80 ppm of aerosolized PAA for 20 min caused 2.3, 1.9, and 0.8 log reductions of E. coli O157:H7, S. Typhimurium, and L. monocytogenes, respectively. Combined treatment of ClO2 gas (10 ppmv) and aerosolized PAA (80 ppm) for 20 min caused 5.4, 5.1, and 4.1 log reductions of E. coli O157:H7, S. Typhimurium, and L. monocytogenes, respectively. E. coli O157:H7, S. Typhimurium, and L. monocytogenes on tomatoes experienced similar reduction patterns to those on spinach leaves. As treatment time increased, most combinations of ClO2 gas and aerosolized PAA showed additive effects in the inactivation of the three pathogens. Combined treatment of ClO2 gas and aerosolized PAA produced injured cells of three pathogens on spinach leaves while generally did not produce injured cells of these pathogens on tomatoes. Combined treatment of ClO2 gas (10 ppmv) and aerosolized PAA (80 ppm) did not significantly (p>0.05) affect the color and texture of samples during 7 days of storage. Copyright © 2015. Published by Elsevier B.V.

Infectious Causes of Cholesteatoma and Treatment of Infected Ossicles prior to Reimplantation by Hydrostatic High-Pressure Inactivation

PubMed Central

Hinz, Rebecca

2015-01-01

Chronic inflammation, which is caused by recurrent infections, is one of the factors contributing to the pathogenesis of cholesteatoma. If reimplantation of autologous ossicles after a surgical intervention is intended, inactivation of planktonic bacteria and biofilms is desirable. High hydrostatic pressure treatment is a procedure, which has been used to inactivate cholesteatoma cells on ossicles. Here we discuss the potential inactivating effect of high hydrostatic pressure on microbial pathogens including biofilms. Recent experimental data suggest an incomplete inactivation at a pressure level, which is tolerable for the bone substance of ossicles and results at least in a considerable reduction of pathogen load. Further studies are necessary to access how far this quantitative reduction of pathogens is sufficient to prevent ongoing chronic infections, for example, due to forming of biofilms. PMID:25705686

Mycobacterium leprae: genes, pseudogenes and genetic diversity

PubMed Central

Singh, Pushpendra; Cole, Stewart T

2011-01-01

Leprosy, which has afflicted human populations for millenia, results from infection with Mycobacterium leprae, an unculturable pathogen with an exceptionally long generation time. Considerable insight into the biology and drug resistance of the leprosy bacillus has been obtained from genomics. M. leprae has undergone reductive evolution and pseudogenes now occupy half of its genome. Comparative genomics of four different strains revealed remarkable conservation of the genome (99.995% identity) yet uncovered 215 polymorphic sites, mainly single nucleotide polymorphisms, and a handful of new pseudogenes. Mapping these polymorphisms in a large panel of strains defined 16 single nucleotide polymorphism-subtypes that showed strong geographical associations and helped retrace the evolution of M. leprae. PMID:21162636

40 CFR 503.10 - Applicability.

Code of Federal Regulations, 2011 CFR

2011-07-01

... pathogen requirements in § 503.32(a); and one of the vector attraction reduction requirements in § 503.33(b... of the vector attraction reduction requirements in § 503.33(b)(1) through (b)(8). (2) The Regional... requirements in § 503.32(a); and one of the vector attraction reduction requirements in § 503.33(b)(1) through...

Research Update - Intervention Technologies for Enhancing the Safety and Security of Fresh and Minimally Processed Produce and Solid Plant-Derived Foods

USDA-ARS?s Scientific Manuscript database

The produce safety research objectives of Research Project 1935-41420-011 are to 1) understand pathogen microbial ecology and its effects on decontamination efficacy; 2) develop biological-based intervention strategies for pathogen reduction; and 3) develop new effective chemical and physical decont...

Minimal heat treatments reduce Shiga toxin-producing Escherichia coli (STEC)and Salmonella on the surface of fresh beef

USDA-ARS?s Scientific Manuscript database

Washing and sanitizing agents have been effective in reducing bacterial populations and pathogen presence on carcasses. Thermal interventions consistently provide the greatest pathogen reductions and can be applied during slaughter in a number of different forms, either as a whole carcass wash, or t...

New wash aid T-128 improves efficacy of chlorine against cross contamination by bacterial pathogens in fresh-cut lettuce processing

USDA-ARS?s Scientific Manuscript database

Chlorinated water is widely used as the primary anti-microbial intervention during fresh-cut produce processing. Free chlorine in chlorinated water can provide effective reduction of potential contaminations by microbial pathogens, and, more importantly, effectively prevent cross contamination of p...

Assessment of the pathogenicity of cell-culture-adapted Newcastle disease virus strain Komarov.

PubMed

Visnuvinayagam, Sivam; Thangavel, K; Lalitha, N; Malmarugan, S; Sukumar, Kuppannan

2015-01-01

Newcastle disease vaccines hitherto in vogue are produced from embryonated chicken eggs. Egg-adapted mesogenic vaccines possess several drawbacks such as paralysis and mortality in 2-week-old chicks and reduced egg production in the egg-laying flock. Owing to these possible drawbacks, we attempted to reduce the vaccine virulence for safe vaccination by adapting the virus in a chicken embryo fibroblast cell culture (CEFCC) system. Eighteen passages were carried out by CEFCC, and the pathogenicity was assessed on the basis of the mean death time, intracerebral pathogenicity index, and intravenous pathogenicity index, at equal passage intervals. Although the reduction in virulence demonstrated with increasing passage levels in CEFCC was encouraging, 20% of the 2-week-old birds showed paralytic symptoms with the virus vaccine from the 18(th)(final) passage. Thus, a tissue-culture-adapted vaccine would demand a few more passages by CEFCC in order to achieve a complete reduction in virulence for use as a safe and effective vaccine, especially among younger chicks. Moreover, it can be safely administered even to unprimed 8-week-old birds.

The oxygen reduction pathway and heat shock stress response are both required for Entamoeba histolytica pathogenicity.

PubMed

Olivos-García, Alfonso; Saavedra, Emma; Nequiz, Mario; Santos, Fabiola; Luis-García, Erika Rubí; Gudiño, Marco; Pérez-Tamayo, Ruy

2016-05-01

Several species belonging to the genus Entamoeba can colonize the mouth or the human gut; however, only Entamoeba histolytica is pathogenic to the host, causing the disease amoebiasis. This illness is responsible for one hundred thousand human deaths per year worldwide, affecting mainly underdeveloped countries. Throughout its entire life cycle and invasion of human tissues, the parasite is constantly subjected to stress conditions. Under in vitro culture, this microaerophilic parasite can tolerate up to 5 % oxygen concentrations; however, during tissue invasion the parasite has to cope with the higher oxygen content found in well-perfused tissues (4-14 %) and with reactive oxygen and nitrogen species derived from both host and parasite. In this work, the role of the amoebic oxygen reduction pathway (ORP) and heat shock response (HSP) are analyzed in relation to E. histolytica pathogenicity. The data suggest that in contrast with non-pathogenic E. dispar, the higher level of ORP and HSPs displayed by E. histolytica enables its survival in tissues by diminishing and detoxifying intracellular oxidants and repairing damaged proteins to allow metabolic fluxes, replication and immune evasion.

Edible apple film wraps containing plant antimicrobials inactivate foodborne pathogens on meat and poultry products.

PubMed

Ravishankar, Sadhana; Zhu, Libin; Olsen, Carl W; McHugh, Tara H; Friedman, Mendel

2009-10-01

Apple-based edible films containing plant antimicrobials were evaluated for their activity against pathogenic bacteria on meat and poultry products. Salmonella enterica or E. coli O157:H7 (10(7) CFU/g) cultures were surface inoculated on chicken breasts and Listeria monocytogenes (10(6) CFU/g) on ham. The inoculated products were then wrapped with edible films containing 3 concentrations (0.5%, 1.5%, and 3%) of cinnamaldehyde or carvacrol. Following incubation at either 23 or 4 degrees C for 72 h, samples were stomached in buffered peptone water, diluted, and plated for enumeration of survivors. The antimicrobial films exhibited concentration-dependent activities against the pathogens tested. At 23 degrees C on chicken breasts, films with 3% antimicrobials showed the highest reductions (4.3 to 6.8 log CFU/g) of both S. enterica and E. coli O157:H7. Films with 1.5% and 0.5% antimicrobials showed 2.4 to 4.3 and 1.6 to 2.8 log reductions, respectively. At 4 degrees C, carvacrol exhibited greater activity than did cinnamaldehyde. Films with 3%, 1.5%, and 0.5% carvacrol reduced the bacterial populations by about 3, 1.6 to 3, and 0.8 to 1 logs, respectively. Films with 3% and 1.5% cinnamaldehyde induced 1.2 to 2.8 and 1.2 to 1.3 log reductions, respectively. For L. monocytogenes on ham, carvacrol films induced greater reductions than did cinnamaldehyde films at all concentrations tested. In general, the reduction of L. monocytogenes on ham at 23 degrees C was greater than at 4 degrees C. Added antimicrobials had minor effects on physical properties of the films. The results suggest that the food industry and consumers could use these films as wrappings to control surface contamination by foodborne pathogenic microorganisms.

Chemical composition and antifungal activity of Arnica longifolia, Aster hesperius, and Chrysothamnus nauseosus essential oils.

PubMed

Tabanca, Nurhayat; Demirci, Betul; Crockett, Sara L; Başer, Kemal Hüsnü Can; Wedge, David E

2007-10-17

Essential oils from three different Asteraceae obtained by hydrodistillation of aerial parts were analyzed by gas chromatography (GC) and gas chromatography-mass spectrometry (GC/MS). Main compounds obtained from each taxon were found as follows: Arnica longifolia carvacrol 37.3%, alpha-bisabolol 8.2%; Aster hesperius hexadecanoic acid 29.6%, carvacrol 15.2%; and Chrysothamnus nauseosus var. nauseosus beta-phellandrene 22.8% and beta-pinene 19.8%. Essential oils were also evaluated for their antimalarial and antimicrobial activity against human pathogens, and antifungal activities against plant pathogens. No antimalarial and antimicrobial activities against human pathogens were observed. Direct bioautography demonstrated antifungal activity of the essential oils obtained from three Asteraceae taxa and two pure compounds, carvacrol and beta-bisabolol, to the plant pathogens Colletotrichum acutatum, C. fragariae and C. gloeosporioides. Subsequent evaluation of antifungal compounds using a 96-well micro-dilution broth assay indicated that alpha-bisabolol showed weak growth inhibition of the plant pathogen Botrytis cinerea after 72 h.

Bioactivity-Guided Metabolite Profiling of Feijoa ( Acca sellowiana) Cultivars Identifies 4-Cyclopentene-1,3-dione as a Potent Antifungal Inhibitor of Chitin Synthesis.

PubMed

Mokhtari, Mona; Jackson, Michael D; Brown, Alistair S; Ackerley, David F; Ritson, Nigel J; Keyzers, Robert A; Munkacsi, Andrew B

2018-06-06

Pathogenic fungi continue to develop resistance against current antifungal drugs. To explore the potential of agricultural waste products as a source of novel antifungal compounds, we obtained an unbiased GC-MS profile of 151 compounds from 16 commercial and experimental cultivars of feijoa peels. Multivariate analysis correlated 93% of the compound profiles with antifungal bioactivities. Of the 18 compounds that significantly correlated with antifungal activity, 5 had not previously been described from feijoa. Two novel cultivars were the most bioactive, and the compound 4-cyclopentene-1,3-dione, detected in these cultivars, was potently antifungal (IC 50 = 1-2 μM) against human-pathogenic Candida species. Haploinsufficiency and fluorescence microscopy analyses determined that the synthesis of chitin, a fungal-cell-wall polysaccharide, was the target of 4-cyclopentene-1,3-dione. This fungal-specific mechanism was consistent with a 22-70-fold reduction in antibacterial activity. Overall, we identified the agricultural waste product of specific cultivars of feijoa peels as a source of potential high-value antifungal compounds.

Efficient reduction of pathogenic and spoilage microorganisms from apple cider by combining microfiltration with UV treatment.

PubMed

Zhao, Dongjun; Barrientos, Jessie Usaga; Wang, Qing; Markland, Sarah M; Churey, John J; Padilla-Zakour, Olga I; Worobo, Randy W; Kniel, Kalmia E; Moraru, Carmen I

2015-04-01

Thermal pasteurization can achieve the U. S. Food and Drug Administration-required 5-log reduction of pathogenic Escherichia coli O157:H7 and Cryptosporidium parvum in apple juice and cider, but it can also negatively affect the nutritional and organoleptic properties of the treated products. In addition, thermal pasteurization is only marginally effective against the acidophilic, thermophilic, and spore-forming bacteria Alicyclobacillus spp., which is known to cause off-flavors in juice products. In this study, the efficiency of a combined microfiltration (MF) and UV process as a nonthermal treatment for the reduction of pathogenic and nonpathogenic E. coli, C. parvum, and Alicyclobacillus acidoterrestris from apple cider was investigated. MF was used to physically remove suspended solids and microorganisms from apple cider, thus enhancing the effectiveness of UV and allowing a lower UV dose to be used. MF, with ceramic membranes (pore sizes, 0.8 and 1.4 μm), was performed at a temperature of 10 °C and a transmembrane pressure of 155 kPa. The subsequent UV treatment was conducted using at a low UV dose of 1.75 mJ/cm(2). The combined MF and UV achieved more than a 5-log reduction of E. coli, C. parvum, and A. acidoterrestris. MF with the 0.8-μm pore size performed better than the 1.4-μm pore size on removal of E. coli and A. acidoterrestris. The developed nonthermal hurdle treatment has the potential to significantly reduce pathogens, as well as spores, yeasts, molds, and protozoa in apple cider, and thus help juice processors improve the safety and quality of their products.

Nitric Oxide in the Offensive Strategy of Fungal and Oomycete Plant Pathogens

PubMed Central

Arasimowicz-Jelonek, Magdalena; Floryszak-Wieczorek, Jolanta

2016-01-01

In the course of evolutionary changes pathogens have developed many invasion strategies, to which the host organisms responded with a broad range of defense reactions involving endogenous signaling molecules, such as nitric oxide (NO). There is evidence that pathogenic microorganisms, including two most important groups of eukaryotic plant pathogens, also acquired the ability to synthesize NO via non-unequivocally defined oxidative and/or reductive routes. Although the both kingdoms Chromista and Fungi are remarkably diverse, the experimental data clearly indicate that pathogen-derived NO is an important regulatory molecule controlling not only developmental processes, but also pathogen virulence and its survival in the host. An active control of mitigation or aggravation of nitrosative stress within host cells seems to be a key determinant for the successful invasion of plant pathogens representing different lifestyles and an effective mode of dispersion in various environmental niches. PMID:26973690

Electron beam inactivation of selected microbial pathogens and indicator organisms in aerobically and anaerobically digested sewage sludge.

PubMed

Praveen, Chandni; Jesudhasan, Palmy R; Reimers, Robert S; Pillai, Suresh D

2013-09-01

Microbial pathogens in municipal sewage sludges need to be inactivated prior to environmental disposal. The efficacy of high energy (10 MeV) e-beam irradiation to inactivate a variety of selected microbial pathogens and indicator organisms in aerobically and anaerobically digested sewage sludge was evaluated. Both bacterial and viral pathogens and indicator organisms are susceptible to e-beam irradiation. However, as expected there was a significant difference in their respective e-beam irradiation sensitivity. Somatic coliphages, bacterial endospores and enteric viruses were more resistant compared to bacterial pathogens. The current US EPA mandated 10 kGy minimum dose was capable of achieving significant reduction of both bacterial and viral pathogens. Somatic coliphages can be used as a microbial indicator for monitoring e-beam processes in terms of pathogen inactivation in sewage sludges. Copyright © 2013 Elsevier Ltd. All rights reserved.

Biological activity of green-synthesized silver nanoparticles depends on the applied natural extracts: a comprehensive study.

PubMed

Rónavári, Andrea; Kovács, Dávid; Igaz, Nóra; Vágvölgyi, Csaba; Boros, Imre Miklós; Kónya, Zoltán; Pfeiffer, Ilona; Kiricsi, Mónika

2017-01-01

Due to obvious disadvantages of the classical chemical methods, green synthesis of metallic nanoparticles has attracted tremendous attention in recent years. Numerous environmentally benign synthesis methods have been developed yielding nanoparticles via low-cost, eco-friendly, and simple approaches. In this study, our aim was to determine the suitability of coffee and green tea extracts in green synthesis of silver nanoparticles as well as to compare the performance of the obtained materials in different biological systems. We successfully produced silver nanoparticles (C-AgNP and GT-AgNP) using coffee and green tea extracts; moreover, based on our comprehensive screening, we delineated major differences in the biological activity of C-AgNPs and GT-AgNPs. Our results indicate that although GT-AgNPs exhibited excellent antimicrobial activity against all the examined microbial pathogens, these particles were also highly toxic to mammalian cells, which limits their potential applications. On the contrary, C-AgNPs manifested substantial inhibitory action on the tested microbes but were nontoxic to human and mouse cells, indicating an outstanding capacity to discriminate between potential pathogens and mammalian cells. These results clearly show that the various green materials used for stabilization and for reduction of metal ions have a defining role in determining and fine-tuning the biological activity of the obtained nanoparticles.

Biological activity of green-synthesized silver nanoparticles depends on the applied natural extracts: a comprehensive study

PubMed Central

Rónavári, Andrea; Kovács, Dávid; Igaz, Nóra; Vágvölgyi, Csaba; Boros, Imre Miklós; Kónya, Zoltán; Pfeiffer, Ilona; Kiricsi, Mónika

2017-01-01

Due to obvious disadvantages of the classical chemical methods, green synthesis of metallic nanoparticles has attracted tremendous attention in recent years. Numerous environmentally benign synthesis methods have been developed yielding nanoparticles via low-cost, eco-friendly, and simple approaches. In this study, our aim was to determine the suitability of coffee and green tea extracts in green synthesis of silver nanoparticles as well as to compare the performance of the obtained materials in different biological systems. We successfully produced silver nanoparticles (C-AgNP and GT-AgNP) using coffee and green tea extracts; moreover, based on our comprehensive screening, we delineated major differences in the biological activity of C-AgNPs and GT-AgNPs. Our results indicate that although GT-AgNPs exhibited excellent antimicrobial activity against all the examined microbial pathogens, these particles were also highly toxic to mammalian cells, which limits their potential applications. On the contrary, C-AgNPs manifested substantial inhibitory action on the tested microbes but were nontoxic to human and mouse cells, indicating an outstanding capacity to discriminate between potential pathogens and mammalian cells. These results clearly show that the various green materials used for stabilization and for reduction of metal ions have a defining role in determining and fine-tuning the biological activity of the obtained nanoparticles. PMID:28184158

FgBud3, a Rho4-Interacting Guanine Nucleotide Exchange Factor, Is Involved in Polarity Growth, Cell Division and Pathogenicity of Fusarium graminearum.

PubMed

Zhang, Chengkang; Luo, Zenghong; He, Dongdong; Su, Li; Yin, Hui; Wang, Guo; Liu, Hong; Rensing, Christopher; Wang, Zonghua

2018-01-01

Rho GTPases are signaling macromolecules that are associated with developmental progression and pathogenesis of Fusarium graminearum . Generally, enzymatic activities of Rho GTPases are regulated by Rho GTPase guanine nucleotide exchange factors (RhoGEFs). In this study, we identified a putative RhoGEF encoding gene ( FgBUD3 ) in F. graminearum database and proceeded further by using a functional genetic approach to generate FgBUD3 targeted gene deletion mutant. Phenotypic analysis results showed that the deletion of FgBUD3 caused severe reduction in growth of FgBUD3 mutant generated during this study. We also observed that the deletion of FgBUD3 completely abolished sexual reproduction and triggered the production of abnormal asexual spores with nearly no septum in ΔFgbud3 strain. Further results obtained from infection assays conducted during this research revealed that the FgBUD3 defective mutant lost its pathogenicity on wheat and hence, suggests FgBud3 plays an essential role in the pathogenicity of F. graminearum . Additional, results derived from yeast two-hybrid assays revealed that FgBud3 strongly interacted with FgRho4 compared to the interaction with FgRho2, FgRho3, and FgCdc42. Moreover, we found that FgBud3 interacted with both GTP-bound and GDP-bound form of FgRho4. From these results, we subsequently concluded that, the Rho4-interacting GEF protein FgBud3 crucially promotes vegetative growth, asexual and sexual development, cell division and pathogenicity in F. graminearum .

Evaluation of a microwave based reactor for the treatment of blackwater sludge

PubMed Central

Mawioo, Peter M.; Rweyemamu, Audax; Garcia, Hector A.; Hooijmans, Christine M.; Brdjanovic, Damir

2016-01-01

A laboratory-scale microwave (MW) unit was applied to treat fresh blackwater sludge that represented fecal sludge (FS) produced at heavily used toilet facilities. The sludge was exposed to MW irradiation at different power levels and for various durations. Variables such as sludge volume and pathogen reduction were observed. The results demonstrated that the MW is a rapid and efficient technology that can reduce the sludge volume by over 70% in these experimental conditions. The concentration of bacterial pathogenic indicator E. coli also decreased to below the analytical detection levels. Furthermore, the results indicated that the MW operational conditions including radiation power and contact time can be varied to achieve the desired sludge volume and pathogen reduction. MW technology can be further explored for the potential scaling-up as an option for rapid treatment of FS from intensively used sanitation facilities such as in emergency situations. PMID:26799809

Biofiltration for stormwater harvesting: Comparison of Campylobacter spp. and Escherichia coli removal under normal and challenging operational conditions

NASA Astrophysics Data System (ADS)

Chandrasena, G. I.; Deletic, A.; McCarthy, D. T.

2016-06-01

Knowledge of pathogen removal in stormwater biofilters (also known as stormwater bioretention systems or rain gardens) has predominately been determined using bacterial indicators, and the removal of reference pathogens in these systems has rarely been investigated. Furthermore, current understanding of indicator bacteria removal in these systems is largely built upon laboratory-scale work. This paper examines whether indicator organism removal from urban stormwater using biofilters in laboratory settings are representative of the removal of pathogens in field conditions, by studying the removal of Escherichia coli (a typical indicator microorganism) and Campylobacter spp. (a typical reference pathogen) from urban stormwater by two established field-scale biofilters. It was found that E. coli log reduction was higher than that of Campylobacter spp. in both biofilters, and that there was no correlation between E. coli and Campylobacter spp. log removal performance. This confirms that E. coli behaves significantly differently to this reference pathogen, reinforcing that single organisms should not be employed to understand faecal microorganism removal in urban stormwater treatment systems. The average reduction in E. coli from only one of the tested biofilters was able to meet the log reduction targets suggested in the current Australian stormwater harvesting guidelines for irrigating sports fields and golf courses. The difference in the performance of the two biofilters is likely a result of a number of design and operational factors; the most important being that the biofilter that did not meet the guidelines was tested using extremely high influent volumes and microbial concentrations, and long antecedent dry weather periods. As such, the E. coli removal performances identified in this study confirmed laboratory findings that inflow concentration and antecedent dry period impact overall microbial removal. In general, this paper emphasizes the need for the validation of stormwater harvesting systems, namely, the testing of treatment systems under challenging operational conditions using multiple indicators and reference pathogens.

Reduction of Escherichia coli O157:H7 on produce by use of electrolyzed water under simulated food service operation conditions.

PubMed

Pangloli, Philipus; Hung, Yen-Con; Beuchat, Larry R; King, C Harold; Zhao, Zhi-Hui

2009-09-01

Treatment of fresh fruits and vegetables with electrolyzed water (EW) has been shown to kill or reduce foodborne pathogens. We evaluated the efficacy of EW in killing Escherichia coli O157:H7 on iceberg lettuce, cabbage, lemons, and tomatoes by using washing and/or chilling treatments simulating those followed in some food service kitchens. Greatest reduction levels on lettuce were achieved by sequentially washing with 14-A (amperage) acidic EW (AcEW) for 15 or 30 s followed by chilling in 16-A AcEW for 15 min. This procedure reduced the pathogen by 2.8 and 3.0 log CFU per leaf, respectively, whereas washing and chilling with tap water reduced the pathogen by 1.9 and 2.4 log CFU per leaf. Washing cabbage leaves for 15 or 30 s with tap water or 14-A AcEW reduced the pathogen by 2.0 and 3.0 log CFU per leaf and 2.5 to 3.0 log CFU per leaf, respectively. The pathogen was reduced by 4.7 log CFU per lemon by washing with 14-A AcEW and 4.1 and 4.5 log CFU per lemon by washing with tap water for 15 or 30 s. A reduction of 5.3 log CFU per lemon was achieved by washing with 14-A alkaline EW for 15 s prior to washing with 14-A AcEW for 15 s. Washing tomatoes with tap water or 14-A AcEW for 15 s reduced the pathogen by 6.4 and 7.9 log CFU per tomato, respectively. Application of AcEW using procedures mimicking food service operations should help minimize cross-contamination and reduce the risk of E. coli O157:H7 being present on produce at the time of consumption.

Preliminary evaluation of pathogenic bacteria loading on organic Municipal Solid Waste compost and vermicompost.

PubMed

Soobhany, Nuhaa

2018-01-15

The use of composts or vermicomposts derived from organic fraction of Municipal Solid Waste (OFMSW) brought about certain disagreement in terms of high level of bacterial pathogens, thereby surpassing the legal restrictions. This preliminary study was undertaken to compare the evolution of pathogenic bacteria on OFMSW compost against vermicompost (generated by Eudrilus eugeniae) with promises of achieving sanitation goals. Analysis to quality data showed that OFMSW vermicomposting caused a moderately higher reduction in total coliforms in contrast to composting. E. coli in OFMSW composts was found to be in the range of 4.72-4.96 log 10  CFU g -1 whilst on a clear contrary, E. coli was undetectable in the final vermicomposts (6.01-6.14 logs of reduction) which might be explained by the involvement of the digestive processes in worms' guts. Both OFMSW composts and vermicomposts generated Salmonella-free products which were acceptable for agricultural usage and soil improvement. In comparison to compost, the analysis of this research indicated that earthworm activity can effectively destroy bacterial pathogenic load in OFMSW vermicomposts. But still, this study necessitates extra research in order to comprehend the factors that direct pathogenic bacteria in vermicomposting and earthworm-free decomposition systems. Copyright © 2017 Elsevier Ltd. All rights reserved.

Biological and Physicochemical Wastewater Treatment Processes Reduce the Prevalence of Virulent Escherichia coli

PubMed Central

Biswal, Basanta Kumar; Mazza, Alberto; Masson, Luke; Gehr, Ronald

2013-01-01

Effluents discharged from wastewater treatment plants are possible sources of pathogenic bacteria, including Escherichia coli, in the freshwater environment, and determining the possible selection of pathogens is important. This study evaluated the impact of activated sludge and physicochemical wastewater treatment processes on the prevalence of potentially virulent E. coli. A total of 719 E. coli isolates collected from four municipal plants in Québec before and after treatment were characterized by using a customized DNA microarray to determine the impact of treatment processes on the frequency of specific pathotypes and virulence genes. The percentages of potentially pathogenic E. coli isolates in the plant influents varied between 26 and 51%, and in the effluents, the percentages were 14 to 31%, for a reduction observed at all plants ranging between 14 and 45%. Pathotypes associated with extraintestinal pathogenic E. coli (ExPEC) were the most abundant at three of the four plants and represented 24% of all isolates, while intestinal pathogenic E. coli pathotypes (IPEC) represented 10% of the isolates. At the plant where ExPEC isolates were not the most abundant, a large number of isolates were classified as both ExPEC and IPEC; overall, 6% of the isolates were classified in both groups, with the majority being from the same plant. The reduction of the proportion of pathogenic E. coli could not be explained by the preferential loss of one virulence gene or one type of virulence factor; however, the quinolone resistance gene (qnrS) appears to enhance the loss of virulence genes, suggesting a mechanism involving the loss of pathogenicity islands. PMID:23160132

FNR Regulates the Expression of Important Virulence Factors Contributing to the Pathogenicity of Avian Pathogenic Escherichia coli

PubMed Central

Barbieri, Nicolle L.; Vande Vorde, Jessica A.; Baker, Alison R.; Horn, Fabiana; Li, Ganwu; Logue, Catherine M.; Nolan, Lisa K.

2017-01-01

Avian pathogenic Escherichia coli (APEC) is the etiologic agent of colibacillosis, an important cause of morbidity and mortality in poultry. Though, many virulence factors associated with APEC pathogenicity are known, their regulation remains unclear. FNR (fumarate and nitrate reduction) is a well-known global regulator that works as an oxygen sensor and has previously been described as a virulence regulator in bacterial pathogens. The goal of this study was to examine the role of FNR in the regulation of APEC virulence factors, such as Type I fimbriae, and processes such as adherence and invasion, type VI secretion, survival during oxidative stress, and growth in iron-restricted environments. To accomplish this goal, APEC O1, a well-characterized, highly virulent, and fully sequenced strain of APEC harboring multiple virulence mechanisms, some of which are plasmid-linked, was compared to its FNR mutant for expression of various virulence traits. Deletion of FNR was found to affect APEC O1's adherence, invasion and expression of ompT, a plasmid-encoded outer membrane protein, type I fimbriae, and aatA, encoding an autotransporter. Indeed, the fnr− mutant showed an 8-fold reduction in expression of type I fimbriae and a highly significant (P < 0.0001) reduction in expression of fimA, ompT (plasmid-borne), and aatA. FNR was also found to regulate expression of the type VI secretion system, affecting the expression of vgrG. Further, FNR was found to be important to APEC O1's growth in iron-deficient media and survival during oxidative stress with the mutant showing a 4-fold decrease in tolerance to oxidative stress, as compared to the wild type. Thus, our results suggest that FNR functions as an important regulator of APEC virulence. PMID:28690981

Efficacy of antimicrobials for the disinfection of pathogen contaminated green bell pepper and of consumer cleaning methods for the decontamination of knives.

PubMed

Perez, Keila L; Lucia, Lisa M; Cisneros-Zevallos, Luis; Castillo, Alejandro; Taylor, T Matthew

2012-05-01

While there is strong focus on eliminating pathogens from produce at a commercial level, consumers can employ simple methods to achieve additional pathogen reductions in the domestic kitchen. To determine the ability of antimicrobials to decontaminate peppers, samples of green bell pepper were inoculated with Salmonella enterica and Escherichia coli O157:H7 and then immersed in 3% (v/v) hydrogen peroxide (H₂O₂), 2.5% (v/v) acetic acid (AA), 70% (v/v) ethyl alcohol (EtOH), or sterile distilled water (SDW). The potential for transfer of pathogens from contaminated peppers to other non-contaminated produce items, and the effect of knife disinfection in preventing this cross contamination, were also tested. Knife disinfection procedures were evaluated by chopping inoculated peppers into 1 cm² pieces with kitchen knives. Experimental knives were then treated by either no treatment (control), wiping with a dry sterile cotton towel, rinsing under running warm water for 5 or 10s, or applying a 1% (v/v) lauryl sulfate-based detergent solution followed by rinsing with warm running water for 10s. Following disinfection treatment, knives were used to slice cucumbers. Exposure to H₂O₂ for 5 min and EtOH for 1 min resulted in reductions of 1.3±0.3 log₁₀ CFU/cm² for both pathogens. A 5 min exposure to AA resulted in a reduction of S. enterica of 1.0±0.7 log₁₀ CFU/cm² and E. coli of 0.7±0.8 log₁₀ CFU/cm². No differences (p ≥ 0.05) were found between numbers of pathogens on knives and numbers of pathogens transferred to cucumber slices, suggesting that organisms remaining on knife surfaces were transferred to cucumbers during slicing. Findings suggest that EtOH and H₂O₂ may be effective antimicrobials for in-home decontamination of peppers, and that use of detergent and warm water is effective for decontamination of implements used during meal preparation. Published by Elsevier B.V.

Evaluating bacterial pathogen DNA preservation in museum osteological collections

PubMed Central

Barnes, Ian; Thomas, Mark G

2005-01-01

Reports of bacterial pathogen DNA sequences obtained from archaeological bone specimens raise the possibility of greatly improving our understanding of the history of infectious diseases. However, the survival of pathogen DNA over long time periods is poorly characterized, and scepticism remains about the reliability of these data. In order to explore the survival of bacterial pathogen DNA in bone specimens, we analysed samples from 59 eighteenth and twentieth century individuals known to have been infected with either Mycobacterium tuberculosis or Treponema pallidum. No reproducible evidence of surviving pathogen DNA was obtained, despite the use of extraction and PCR-amplification methods determined to be highly sensitive. These data suggest that previous studies need to be interpreted with caution, and we propose that a much greater emphasis is placed on understanding how pathogen DNA survives in archaeological material, and how its presence can be properly verified and used. PMID:16608682

Extraction of Hemocytes from Drosophila melanogaster Larvae for Microbial Infection and Analysis.

PubMed

Hiroyasu, Aoi; DeWitt, David C; Goodman, Alan G

2018-05-24

During the pathogenic infection of Drosophila melanogaster, hemocytes play an important role in the immune response throughout the infection. Thus, the goal of this protocol is to develop a method to visualize the pathogen invasion in a specific immune compartment of flies, namely hemocytes. Using the method presented here, up to 3 × 10 6 live hemocytes can be obtained from 200 Drosophila 3 rd instar larvae in 30 min for ex vivo infection. Alternatively, hemocytes can be infected in vivo through injection of 3 rd instar larvae followed by hemocyte extraction up to 24 h post-infection. These infected primary cells were fixed, stained, and imaged using confocal microscopy. Then, 3D representations were generated from the images to definitively show pathogen invasion. Additionally, high-quality RNA for qRT-PCR can be obtained for the detection of pathogen mRNA following infection, and sufficient protein can be extracted from these cells for Western blot analysis. Taken together, we present a method for definite reconciliation of pathogen invasion and confirmation of infection using bacterial and viral pathogen types and an efficient method for hemocyte extraction to obtain enough live hemocytes from Drosophila larvae for ex vivo and in vivo infection experiments.

Estimation of norovirus infection risks to consumers of wastewater-irrigated food crops eaten raw.

PubMed

Mara, Duncan; Sleigh, Andrew

2010-03-01

A quantitative microbial risk analysis-Monte Carlo method was used to estimate norovirus infection risks to consumers of wastewater-irrigated lettuce. Using the same assumptions as used in the 2006 WHO guidelines for the safe use of wastewater in agriculture, a norovirus reduction of 6 log units was required to achieve a norovirus infection risk of approximately 10(-3) per person per year (pppy), but for a lower consumption of lettuce (40-48 g per week vs. 350 g per week) the required reduction was 5 log units. If the tolerable additional disease burden is increased from a DALY (disability-adjusted life year) loss of 10(-6) pppy (the value used in the WHO guidelines) to 10(-5) pppy, the required pathogen reduction is one order of magnitude lower. Reductions of 4-6 log units can be achieved by very simple partial treatment (principally settling to achieve a 1-log unit reduction) supplemented by very reliable post-treatment health-protection control measures such as pathogen die-off (1-2 log units), produce washing in cold water (1 log unit) and produce disinfection (3 log units).

Small-scale shifting mosaics of two dominant grassland species: the possible role of soil-borne pathogens.

PubMed

Olff, H; Hoorens, B; de Goede, R G M; van der Putten, W H; Gleichman, J M

2000-10-01

We analyzed the dynamics of dominant plant species in a grazed grassland over 17 years, and investigated whether local shifts in these dominant species, leading to vegetation mosaics, could be attributed to interactions between plants and soil-borne pathogens. We found that Festuca rubra and Carex arenaria locally alternated in abundance, with different sites close together behaving out of phase, resulting in a shifting mosaic. The net effect of killing all soil biota on the growth of these two species was investigated in a greenhouse experiment using gamma radiation, controlling for possible effects of sterilization on soil chemistry. Both plant species showed a strong net positive response to soil sterilization, indicating that pathogens (e.g., nematodes, pathogenic fungi) outweighed the effect of mutualists (e.g., mycorrhizae). This positive growth response towards soil sterilization appeared not be due to effects of sterilization on soil chemistry. Growth of Carex was strongly reduced by soil-borne pathogens (86% reduction relative to its growth on sterilized soil) on soil from a site where this species decreased during the last decade (and Festuca increased), while it was reduced much less (50%) on soil from a nearby site where it increased in abundance during the last decade. Similarly, Festuca was reduced more (67%) on soil from the site where it decreased (and Carex increased) than on soil from the site where it increased (55%, the site where Carex decreased). Plant-feeding nematodes showed high small-scale variation in densities, and we related this variation to the observed growth reductions in both plant species. Carex growth on unsterilized soil was significantly more reduced at higher densities of plant-feeding nematodes, while the growth reduction in Festuca was independent of plant-feeding nematode densities. At high plant-feeding nematode densities, growth of Carex was reduced more than Festuca, while at low nematode densities the opposite was found. Each plant species thus seems to be affected by different (groups of) soil-borne pathogens. The resulting interaction web of plants and soil-borne pathogens is discussed. We hypothesize that soil disturbances by digging ants and rabbits may explain the small-scale variation in nematode densities, by locally providing "fresh" sand. We conclude that soil-borne pathogens may contribute to plant diversity and spatial mosaics of plants in grasslands.

Effect of temperature on chlorine dioxide inactivation of Escherichia coli O157:H7, Salmonella typhimurium, and Listeria monocytogenes on spinach, tomatoes, stainless steel, and glass surfaces.

PubMed

Park, Sang-Hyun; Kang, Dong-Hyun

2018-06-20

The objective of this study was to evaluate how treatment temperature influences the solubility of ClO 2 gas and the antimicrobial effect of ClO 2 gas against Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes on produce and food contact surfaces. Produce and food contact surfaces inoculated with a combined culture cocktail of three strains each of the three foodborne pathogens were processed in a treatment chamber with 20 ppmv ClO 2 gas at 15 or 25 °C under the same conditions of absolute humidity (11.2-12.3 g/m 3 ) for up to 30 min. As treatment time increased, ClO 2 gas treatment at 15 °C caused significantly more (p < 0.05) inactivation of the three pathogens than treatment at 25 °C. ClO 2 gas treatment at 25 °C for 30 min resulted in 1.15 to 1.54, 1.53 to 1.88, and 1.00 to 1.78 log reductions of the three pathogens on spinach leaves, tomatoes, and stainless steel No.4, respectively. ClO 2 gas treatment at 15 °C for 30 min caused 2.53 to 2.88, 2.82 to 3.23, and 2.37 to 3.03 log reductions of the three pathogens on spinach leaves, tomatoes, and stainless steel No.4, respectively. Treatment with ClO 2 gas at 25 °C for 20 min resulted in 1.88 to 2.31 log reductions of the three pathogens on glass while >5.91 to 6.82 log reductions of these pathogens occurred after 20 min when treated at 15 °C. Residual ClO 2 levels after gas treatment at 15 °C were significantly (p < 0.05) higher than those at 25 °C. The results of this study can help the food processing industry establish optimum ClO 2 gas treatment conditions for maximizing the antimicrobial efficacy of ClO 2 gas. Published by Elsevier B.V.

Prudent Use of Antimicrobials in Exotic Animal Medicine.

PubMed

Broens, Els M; van Geijlswijk, Ingeborg M

2018-05-01

Reduction of antimicrobial use can result in reduction of resistance in commensal bacteria. In exotic animals, information on use of antimicrobials and resistance in commensals and pathogens is scarce. However, use of antimicrobials listed as critically important antimicrobials for human medicine seems high in exotic animals. Ideally, the selection of a therapy should be based on an accurate diagnosis and antimicrobial susceptibility testing. When prescribing antimicrobials based on empiricism, knowledge of the most common pathogens causing specific infections and the antimicrobial spectrum of antimicrobial agents is indispensable. Implementing antimicrobial stewardship promotes the prudent use of antimicrobials in exotic animals. Copyright © 2018 Elsevier Inc. All rights reserved.

Subcontinental impacts of an invasive tree disease on forest structure and dynamics

Treesearch

Jeffrey R. Garnas; Matthew P. Ayres; Andrew M. Liebhold; Celia. Evans

2011-01-01

Introduced pests and pathogens are a major source of disturbance to ecosystems world-wide. The famous examples have produced dramatic reductions in host abundance, including virtual extirpation, but most introductions have more subtle impacts that are hard to quantify but are potentially at least as important due to the pathogens' effects on host reproduction,...

In vivo virulence of MHC-adapted AIDS virus serially-passaged through MHC-mismatched hosts

PubMed Central

Yamamoto, Hiroyuki; Ishii, Hiroshi; Matsuoka, Saori; Mizuta, Kazuta; Sakawaki, Hiromi; Miura, Tomoyuki; Naruse, Taeko K.; Kimura, Akinori

2017-01-01

CD8+ T-cell responses exert strong suppressive pressure on HIV replication and select for viral escape mutations. Some of these major histocompatibility complex class I (MHC-I)-associated mutations result in reduction of in vitro viral replicative capacity. While these mutations can revert after viral transmission to MHC-I-disparate hosts, recent studies have suggested that these MHC-I-associated mutations accumulate in populations and make viruses less pathogenic in vitro. Here, we directly show an increase in the in vivo virulence of an MHC-I-adapted virus serially-passaged through MHC-I-mismatched hosts in a macaque AIDS model despite a reduction in in vitro viral fitness. The first passage simian immunodeficiency virus (1pSIV) obtained 1 year after SIVmac239 infection in a macaque possessing a protective MHC-I haplotype 90-120-Ia was transmitted into 90-120-Ia- macaques, whose plasma 1 year post-infection was transmitted into other 90-120-Ia- macaques to obtain the third passage SIV (3pSIV). Most of the 90-120-Ia-associated mutations selected in 1pSIV did not revert even in 3pSIV. 3pSIV showed lower in vitro viral fitness but induced persistent viremia in 90-120-Ia- macaques. Remarkably, 3pSIV infection in 90-120-Ia+ macaques resulted in significantly higher viral loads and reduced survival compared to wild-type SIVmac239. These results indicate that MHC-I-adapted SIVs serially-transmitted through MHC-I-mismatched hosts can have higher virulence in MHC-I-matched hosts despite their lower in vitro viral fitness. This study suggests that multiply-passaged HIVs could result in loss of HIV-specific CD8+ T cell responses in human populations and the in vivo pathogenic potential of these escaped viruses may be enhanced. PMID:28931083

Reduction of Microbial and Chemical Contaminants in Water Using POU/POE & Mobile Treatment Technologies

EPA Science Inventory

POU/POE may be a cost-effective option for reductions of a particular chemical to achieve water quality compliance under certain situations and given restrictions. Proactive consumers seeking to reduce exposure to potential pathogens, trace chemicals, and nanoparticles not curre...

Improving Pathogen Reduction by Chlorine Wash Prior to Cutting in Fresh-Cut Processing

USDA-ARS?s Scientific Manuscript database

Introduction: Currently, most fresh-cut processing facilities in the United States use chlorinated water or other sanitizer solutions for microbial reduction after lettuce is cut. Freshly cut lettuce releases significant amounts of organic matter that negatively impacts the effectiveness of chlorine...

Pathogen inactivation treatment of plasma and platelet concentrates and their predicted functionality in massive transfusion protocols.

PubMed

Arbaeen, Ahmad F; Schubert, Peter; Serrano, Katherine; Carter, Cedric J; Culibrk, Brankica; Devine, Dana V

2017-05-01

Trauma transfusion packages for hemorrhage control consist of red blood cells, plasma, and platelets at a set ratio. Although pathogen reduction improves the transfusion safety of platelet and plasma units, there is an associated reduction in quality. This study aimed to investigate the impact of riboflavin/ultraviolet light-treated plasma or platelets in transfusion trauma packages composed of red blood cell, plasma, and platelet units in a ratio of 1:1:1 in vitro by modeling transfusion scenarios for trauma patients and assessing function by rotational thromboelastometry. Pathogen-reduced or untreated plasma and buffy coat platelet concentrate units produced in plasma were used in different combinations with red blood cells in trauma transfusion packages. After reconstitution of these packages with hemodiluted blood, the hemostatic functionality was analyzed by rotational thromboelastometry. Hemostatic profiles of pathogen-inactivated buffy coat platelet concentrate and plasma indicated decreased activity compared with their respective controls. Reconstitution of hemodiluted blood (hematocrit = 20%) with packages that contained treated or nontreated components resulted in increased alpha and maximum clot firmness and enhanced clot-formation time. Simulating transfusion scenarios based on 30% blood replacement with a transfusion trauma package resulted in a nonsignificant difference in rotational thromboelastometry parameters between packages containing treated and nontreated blood components (p ≥ 0.05). Effects of pathogen inactivation treatment were evident when the trauma package percentage was 50% or greater and contained both pathogen inactivation-treated plasma and buffy coat platelet concentrate. Rotational thromboelastometry investigations suggest that there is relatively little impact of pathogen inactivation treatment on whole blood clot formation unless large amounts of treated components are used. © 2017 AABB.

Effects of damping-off caused by Rhizoctonia solani anastomosis group 2-1 on roots of wheat and oil seed rape quantified using X-ray Computed Tomography and real-time PCR

PubMed Central

Sturrock, Craig J.; Woodhall, James; Brown, Matthew; Walker, Catherine; Mooney, Sacha J.; Ray, Rumiana V.

2015-01-01

Rhizoctonia solani is a plant pathogenic fungus that causes significant establishment and yield losses to several important food crops globally. This is the first application of high resolution X-ray micro Computed Tomography (X-ray μCT) and real-time PCR to study host–pathogen interactions in situ and elucidate the mechanism of Rhizoctonia damping-off disease over a 6-day period caused by R. solani, anastomosis group (AG) 2-1 in wheat (Triticum aestivum cv. Gallant) and oil seed rape (OSR, Brassica napus cv. Marinka). Temporal, non-destructive analysis of root system architectures was performed using RooTrak and validated by the destructive method of root washing. Disease was assessed visually and related to pathogen DNA quantification in soil using real-time PCR. R. solani AG2-1 at similar initial DNA concentrations in soil was capable of causing significant damage to the developing root systems of both wheat and OSR. Disease caused reductions in primary root number, root volume, root surface area, and convex hull which were affected less in the monocotyledonous host. Wheat was more tolerant to the pathogen, exhibited fewer symptoms and developed more complex root systems. In contrast, R. solani caused earlier damage and maceration of the taproot of the dicot, OSR. Disease severity was related to pathogen DNA accumulation in soil only for OSR, however, reductions in root traits were significantly associated with both disease and pathogen DNA. The method offers the first steps in advancing current understanding of soil-borne pathogen behavior in situ at the pore scale, which may lead to the development of mitigation measures to combat disease influence in the field. PMID:26157449

Effects of damping-off caused by Rhizoctonia solani anastomosis group 2-1 on roots of wheat and oil seed rape quantified using X-ray Computed Tomography and real-time PCR.

PubMed

Sturrock, Craig J; Woodhall, James; Brown, Matthew; Walker, Catherine; Mooney, Sacha J; Ray, Rumiana V

2015-01-01

Rhizoctonia solani is a plant pathogenic fungus that causes significant establishment and yield losses to several important food crops globally. This is the first application of high resolution X-ray micro Computed Tomography (X-ray μCT) and real-time PCR to study host-pathogen interactions in situ and elucidate the mechanism of Rhizoctonia damping-off disease over a 6-day period caused by R. solani, anastomosis group (AG) 2-1 in wheat (Triticum aestivum cv. Gallant) and oil seed rape (OSR, Brassica napus cv. Marinka). Temporal, non-destructive analysis of root system architectures was performed using RooTrak and validated by the destructive method of root washing. Disease was assessed visually and related to pathogen DNA quantification in soil using real-time PCR. R. solani AG2-1 at similar initial DNA concentrations in soil was capable of causing significant damage to the developing root systems of both wheat and OSR. Disease caused reductions in primary root number, root volume, root surface area, and convex hull which were affected less in the monocotyledonous host. Wheat was more tolerant to the pathogen, exhibited fewer symptoms and developed more complex root systems. In contrast, R. solani caused earlier damage and maceration of the taproot of the dicot, OSR. Disease severity was related to pathogen DNA accumulation in soil only for OSR, however, reductions in root traits were significantly associated with both disease and pathogen DNA. The method offers the first steps in advancing current understanding of soil-borne pathogen behavior in situ at the pore scale, which may lead to the development of mitigation measures to combat disease influence in the field.

Review of literature on climate change and forest diseases of western North America

Treesearch

John T. Kliejunas; Brian W. Geils; Jessie Micales Glaeser; Ellen Michaels Goheen; Paul Hennon; Mee-Sook Kim; Harry Kope; Jeff Stone; Rona Sturrock; Susan J. Frankel

2009-01-01

A summary of the literature on relationships between climate and various types of tree diseases, and the potential effects of climate change on pathogens in western North American forests is provided. Climate change generally will lead to reductions in tree health and will improve conditions for some highly damaging pathogens. Sections on abiotic diseases, declines,...

Biochar, Bentonite and Zeolite Supplemented Feeding of Layer Chickens Alters Intestinal Microbiota and Reduces Campylobacter Load

PubMed Central

Prasai, Tanka P.; Walsh, Kerry B.; Bhattarai, Surya P.; Midmore, David J.; Van, Thi T. H.; Moore, Robert J.; Stanley, Dragana

2016-01-01

A range of feed supplements, including antibiotics, have been commonly used in poultry production to improve health and productivity. Alternative methods are needed to suppress pathogen loads and maintain productivity. As an alternative to antibiotics use, we investigated the ability of biochar, bentonite and zeolite as separate 4% feed additives, to selectively remove pathogens without reducing microbial richness and diversity in the gut. Neither biochar, bentonite nor zeolite made any significant alterations to the overall richness and diversity of intestinal bacterial community. However, reduction of some bacterial species, including some potential pathogens was detected. The microbiota of bentonite fed animals were lacking all members of the order Campylobacterales. Specifically, the following operational taxonomic units (OTUs) were absent: an OTU 100% identical to Campylobacter jejuni; an OTU 99% identical to Helicobacter pullorum; multiple Gallibacterium anatis (>97%) related OTUs; Bacteroides dorei (99%) and Clostridium aldenense (95%) related OTUs. Biochar and zeolite treatments had similar but milder effects compared to bentonite. Zeolite amended feed was also associated with significant reduction in the phylum Proteobacteria. All three additives showed potential for the control of major poultry zoonotic pathogens. PMID:27116607

Cleanliness in context: reconciling hygiene with a modern microbial perspective.

PubMed

Vandegrift, Roo; Bateman, Ashley C; Siemens, Kyla N; Nguyen, May; Wilson, Hannah E; Green, Jessica L; Van Den Wymelenberg, Kevin G; Hickey, Roxana J

2017-07-14

The concept of hygiene is rooted in the relationship between cleanliness and the maintenance of good health. Since the widespread acceptance of the germ theory of disease, hygiene has become increasingly conflated with sterilization. In reviewing studies across the hygiene literature (most often hand hygiene), we found that nearly all studies of hand hygiene utilize bulk reduction in bacterial load as a proxy for reduced transmission of pathogenic organisms. This treatment of hygiene may be insufficient in light of recent microbial ecology research, which has demonstrated that humans have intimate and evolutionarily significant relationships with a diverse assemblage of microorganisms (our microbiota). The human skin is home to a diverse and specific community of microorganisms, which include members that exist across the ecological spectrum from pathogen through commensal to mutualist. Most evidence suggests that the skin microbiota is likely of direct benefit to the host and only rarely exhibits pathogenicity. This complex ecological context suggests that the conception of hygiene as a unilateral reduction or removal of microbes has outlived its usefulness. As such, we suggest the explicit definition of hygiene as "those actions and practices that reduce the spread or transmission of pathogenic microorganisms, and thus reduce the incidence of disease."

MoSwi6, an APSES family transcription factor, interacts with MoMps1 and is required for hyphal and conidial morphogenesis, appressorial function, and pathogenicity of Magnaporthe oryzae

PubMed Central

Qi, Zhongqiang; Wang, Qi; Dou, Xianying; Wang, Wei; Zhao, Qian; Lv, Ruili; Zhang, Haifeng; Zheng, Xiaobo; Wang, Ping; Zhang, Zhengguang

2011-01-01

Magnaporthe oryzae MAPK MoMps1 plays a critical role in regulating various developmental processes including cell wall integrity, stress responses, and pathogenicity. To identify potential effectors of MoMps1, we characterized the function of MoSwi6, a homolog of Saccharomyces cerevisiae Swi6 downstream of MAPK Slt2 signaling. MoSwi6 interacted with MoMps1 both in vivo and in vitro, suggesting a possible functional link analogous to Swi6-Slt2 in S. cerevisiae. Targeted gene disruption of MoSWI6 resulted in multiple developmental defects, including reduced hyphal growth, abnormal formation of conidia and appressoria, and impaired appressorium function. The reduction in appressorial turgor pressure also contributed to an attenuation of pathogenicity. The ΔMoswi6 mutant also displayed a defect in cell wall integrity, was hypersensitive to the oxidative stress, and showed significant reduction in transcription and activities of extracellular enzymes including peroxidases and laccases. Collectively, these roles are similar to those of MoMps1, confirming that MoSwi6 functions in the MoMps1 pathway to govern growth, development, and full pathogenicity. PMID:22321443

Natural disturbance reduces disease risk in endangered rainforest frog populations

PubMed Central

Roznik, Elizabeth A.; Sapsford, Sarah J.; Pike, David A.; Schwarzkopf, Lin; Alford, Ross A.

2015-01-01

Natural disturbances can drive disease dynamics in animal populations by altering the microclimates experienced by hosts and their pathogens. Many pathogens are highly sensitive to temperature and moisture, and therefore small changes in habitat structure can alter the microclimate in ways that increase or decrease infection prevalence and intensity in host populations. Here we show that a reduction of rainforest canopy cover caused by a severe tropical cyclone decreased the risk of endangered rainforest frogs (Litoria rheocola) becoming infected by a fungal pathogen (Batrachochytrium dendrobatidis). Reductions in canopy cover increased the temperatures and rates of evaporative water loss in frog microhabitats, which reduced B. dendrobatidis infection risk in frogs by an average of 11–28% in cyclone-damaged areas, relative to unaffected areas. Natural disturbances to the rainforest canopy can therefore provide an immediate benefit to frogs by altering the microclimate in ways that reduce infection risk. This could increase host survival and reduce the probability of epidemic disease outbreaks. For amphibian populations under immediate threat from this pathogen, targeted manipulation of canopy cover could increase the availability of warmer, drier microclimates and therefore tip the balance from host extinction to coexistence. PMID:26294048

Natural disturbance reduces disease risk in endangered rainforest frog populations.

PubMed

Roznik, Elizabeth A; Sapsford, Sarah J; Pike, David A; Schwarzkopf, Lin; Alford, Ross A

2015-08-21

Natural disturbances can drive disease dynamics in animal populations by altering the microclimates experienced by hosts and their pathogens. Many pathogens are highly sensitive to temperature and moisture, and therefore small changes in habitat structure can alter the microclimate in ways that increase or decrease infection prevalence and intensity in host populations. Here we show that a reduction of rainforest canopy cover caused by a severe tropical cyclone decreased the risk of endangered rainforest frogs (Litoria rheocola) becoming infected by a fungal pathogen (Batrachochytrium dendrobatidis). Reductions in canopy cover increased the temperatures and rates of evaporative water loss in frog microhabitats, which reduced B. dendrobatidis infection risk in frogs by an average of 11-28% in cyclone-damaged areas, relative to unaffected areas. Natural disturbances to the rainforest canopy can therefore provide an immediate benefit to frogs by altering the microclimate in ways that reduce infection risk. This could increase host survival and reduce the probability of epidemic disease outbreaks. For amphibian populations under immediate threat from this pathogen, targeted manipulation of canopy cover could increase the availability of warmer, drier microclimates and therefore tip the balance from host extinction to coexistence.

Efficacy of UV-C irradiation for inactivation of food-borne pathogens on sliced cheese packaged with different types and thicknesses of plastic films.

PubMed

Ha, Jae-Won; Back, Kyeong-Hwan; Kim, Yoon-Hee; Kang, Dong-Hyun

2016-08-01

In this study, the efficacy of using UV-C light to inactivate sliced cheese inoculated with Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes and, packaged with 0.07 mm films of polyethylene terephthalate (PET), polyvinylchloride (PVC), polypropylene (PP), and polyethylene (PE) was investigated. The results show that compared with PET and PVC, PP and PE films showed significantly reduced levels of the three pathogens compared to inoculated but non-treated controls. Therefore, PP and PE films of different thicknesses (0.07 mm, 0.10 mm, and 0.13 mm) were then evaluated for pathogen reduction of inoculated sliced cheese samples. Compared with 0.10 and 0.13 mm, 0.07 mm thick PP and PE films did not show statistically significant reductions compared to non-packaged treated samples. Moreover, there were no statistically significant differences between the efficacy of PP and PE films. These results suggest that adjusted PP or PE film packaging in conjunction with UV-C radiation can be applied to control foodborne pathogens in the dairy industry. Copyright © 2016. Published by Elsevier Ltd.

Reinforcing effects of non-pathogenic bacteria and predation risk: from physiology to life history.

PubMed

Janssens, Lizanne; Stoks, Robby

2014-10-01

The important ecological role of predation risk in shaping populations, communities and ecosystems is becoming increasingly clear. In this context, synergistic effects between predation risk and other natural stressors on prey organisms are gaining attention. Although non-pathogenic bacteria can be widespread in aquatic ecosystems, their role in mediating effects of predation risk has been ignored. We here address the hypothesis that non-pathogenic bacteria may reinforce the negative effects of predation risk in larvae of the damselfly Coenagrion puella. We found synergistic effects for all three life history variables studied: mortality increased, growth reductions were magnified and bacterial load was higher when both non-lethal stressors were combined. The combined exposure to the bacterium and predation risk considerably impaired the two key antipredator mechanisms of the damselfly larvae: they no longer reduced their food intake under predation risk and showed a synergistic reduction in escape swimming speed. The reinforcing negative effects on the fitness-related traits could be explained by the observed synergistic effects on food intake, swimming muscle mass, immune function and oxidative damage. These are likely widespread consequences of energetic constraints and increased metabolic rates associated with the fight-or-flight response. We therefore hypothesize that the here documented synergistic interactions with non-pathogenic bacteria may be widespread. Our results highlight the ignored ecological role of non-pathogenic bacteria in reinforcing the negative effects of predation risk on prey organisms.

Zeolite food supplementation reduces abundance of enterobacteria.

PubMed

Prasai, Tanka P; Walsh, Kerry B; Bhattarai, Surya P; Midmore, David J; Van, Thi T H; Moore, Robert J; Stanley, Dragana

2017-01-01

According to the World Health Organisation, antibiotics are rapidly losing potency in every country of the world. Poultry are currently perceived as a major source of pathogens and antimicrobial resistance. There is an urgent need for new and natural ways to control pathogens in poultry and humans alike. Porous, cation rich, aluminosilicate minerals, zeolites can be used as a feed additive in poultry rations, demonstrating multiple productivity benefits. Next generation sequencing of the 16S rRNA marker gene was used to phylogenetically characterize the fecal microbiota and thus investigate the ability and dose dependency of zeolite in terms of anti-pathogenic effects. A natural zeolite was used as a feed additive in laying hens at 1, 2, and 4% w/w for a 23 week period. At the end of this period cloacal swabs were collected to sample faecal microbial communities. A significant reduction in carriage of bacteria within the phylum Proteobacteria, especially in members of the pathogen-rich family Enterobacteriaceae, was noted across all three concentrations of zeolite. Zeolite supplementation of feed resulted in a reduction in the carriage of a number of poultry pathogens without disturbing beneficial bacteria. This effect was, in some phylotypes, correlated with the zeolite concentration. This result is relevant to zeolite feeding in other animal production systems, and for human pathogenesis. Copyright © 2016 Elsevier GmbH. All rights reserved.

Analysis of the Genome and Mobilome of a Dissimilatory Arsenate Reducing Aeromonas sp. O23A Reveals Multiple Mechanisms for Heavy Metal Resistance and Metabolism

PubMed Central

Uhrynowski, Witold; Decewicz, Przemyslaw; Dziewit, Lukasz; Radlinska, Monika; Krawczyk, Pawel S.; Lipinski, Leszek; Adamska, Dorota; Drewniak, Lukasz

2017-01-01

Aeromonas spp. are among the most ubiquitous microorganisms, as they have been isolated from different environmental niches including waters, soil, as well as wounds and digestive tracts of poikilothermic animals and humans. Although much attention has been paid to the pathogenicity of Aeromonads, the role of these bacteria in environmentally important processes, such as transformation of heavy metals, remains to be discovered. Therefore, the aim of this study was a detailed genomic characterization of Aeromonas sp. O23A, the first representative of this genus capable of dissimilatory arsenate reduction. The strain was isolated from microbial mats from the Zloty Stok mine (SW Poland), an environment strongly contaminated with arsenic. Previous physiological studies indicated that O23A may be involved in both mobilization and immobilization of this metalloid in the environment. To discover the molecular basis of the mechanisms behind the observed abilities, the genome of O23A (∼5.0 Mbp) was sequenced and annotated, and genes for arsenic respiration, heavy metal resistance (hmr) and other phenotypic traits, including siderophore production, were identified. The functionality of the indicated gene modules was assessed in a series of minimal inhibitory concentration analyses for various metals and metalloids, as well as mineral dissolution experiments. Interestingly, comparative analyses revealed that O23A is related to a fish pathogen Aeromonas salmonicida subsp. salmonicida A449 which, however, does not carry genes for arsenic respiration. This indicates that the dissimilatory arsenate reduction ability may have been lost during genome reduction in pathogenic strains, or acquired through horizontal gene transfer. Therefore, particular emphasis was placed upon the mobilome of O23A, consisting of four plasmids, a phage, and numerous transposable elements, which may play a role in the dissemination of hmr and arsenic metabolism genes in the environment. The obtained results indicate that Aeromonas sp. O23A is well-adapted to the extreme environmental conditions occurring in the Zloty Stok mine. The analysis of genome encoded traits allowed for a better understanding of the mechanisms of adaptation of the strain, also with respect to its presumable role in colonization and remediation of arsenic-contaminated waters, which may never have been discovered based on physiological analyses alone. PMID:28611742

Analysis of the Genome and Mobilome of a Dissimilatory Arsenate Reducing Aeromonas sp. O23A Reveals Multiple Mechanisms for Heavy Metal Resistance and Metabolism.

PubMed

Uhrynowski, Witold; Decewicz, Przemyslaw; Dziewit, Lukasz; Radlinska, Monika; Krawczyk, Pawel S; Lipinski, Leszek; Adamska, Dorota; Drewniak, Lukasz

2017-01-01

Aeromonas spp. are among the most ubiquitous microorganisms, as they have been isolated from different environmental niches including waters, soil, as well as wounds and digestive tracts of poikilothermic animals and humans. Although much attention has been paid to the pathogenicity of Aeromonads, the role of these bacteria in environmentally important processes, such as transformation of heavy metals, remains to be discovered. Therefore, the aim of this study was a detailed genomic characterization of Aeromonas sp. O23A, the first representative of this genus capable of dissimilatory arsenate reduction. The strain was isolated from microbial mats from the Zloty Stok mine (SW Poland), an environment strongly contaminated with arsenic. Previous physiological studies indicated that O23A may be involved in both mobilization and immobilization of this metalloid in the environment. To discover the molecular basis of the mechanisms behind the observed abilities, the genome of O23A (∼5.0 Mbp) was sequenced and annotated, and genes for arsenic respiration, heavy metal resistance ( hmr ) and other phenotypic traits, including siderophore production, were identified. The functionality of the indicated gene modules was assessed in a series of minimal inhibitory concentration analyses for various metals and metalloids, as well as mineral dissolution experiments. Interestingly, comparative analyses revealed that O23A is related to a fish pathogen Aeromonas salmonicida subsp. salmonicida A449 which, however, does not carry genes for arsenic respiration. This indicates that the dissimilatory arsenate reduction ability may have been lost during genome reduction in pathogenic strains, or acquired through horizontal gene transfer. Therefore, particular emphasis was placed upon the mobilome of O23A, consisting of four plasmids, a phage, and numerous transposable elements, which may play a role in the dissemination of hmr and arsenic metabolism genes in the environment. The obtained results indicate that Aeromonas sp. O23A is well-adapted to the extreme environmental conditions occurring in the Zloty Stok mine. The analysis of genome encoded traits allowed for a better understanding of the mechanisms of adaptation of the strain, also with respect to its presumable role in colonization and remediation of arsenic-contaminated waters, which may never have been discovered based on physiological analyses alone.

40 CFR 503.27 - Recordkeeping.

Code of Federal Regulations, 2010 CFR

2010-07-01

... penalty of law, that the information that will be used to determine compliance with the pathogen... requirements is met) and the vector attraction reduction requirement in (insert one of the vector attraction... met. (iv) A description of how one of the vector attraction reduction requirements in § 503.33 (b)(1...

40 CFR 503.27 - Recordkeeping.

Code of Federal Regulations, 2014 CFR

2014-07-01

... penalty of law, that the information that will be used to determine compliance with the pathogen... requirements is met) and the vector attraction reduction requirement in (insert one of the vector attraction... met. (iv) A description of how one of the vector attraction reduction requirements in § 503.33 (b)(1...

40 CFR 503.27 - Recordkeeping.

Code of Federal Regulations, 2012 CFR

2012-07-01

... penalty of law, that the information that will be used to determine compliance with the pathogen... requirements is met) and the vector attraction reduction requirement in (insert one of the vector attraction... met. (iv) A description of how one of the vector attraction reduction requirements in § 503.33 (b)(1...

40 CFR 503.27 - Recordkeeping.

Code of Federal Regulations, 2013 CFR

2013-07-01

... penalty of law, that the information that will be used to determine compliance with the pathogen... requirements is met) and the vector attraction reduction requirement in (insert one of the vector attraction... met. (iv) A description of how one of the vector attraction reduction requirements in § 503.33 (b)(1...

78 FR 72626 - Notice of Request for Renewal of a Currently Approved Information Collection (Pathogen Reduction...

Federal Register 2010, 2011, 2012, 2013, 2014

2013-12-03

... reduction and Hazard Analysis and Critical Control Point (HACCP) Systems requirements because OMB approval... February 28, 2014. FSIS has established requirements applicable to meat and poultry establishments designed.... coli by slaughter establishments to verify the adequacy of the establishment's process controls for the...

Effect of radiant catalytic ionization on reduction of foodborne pathogens on beef

USDA-ARS?s Scientific Manuscript database

The objective of this study was to evaluate the effect of radiant catalytic ionization (RCI) on reduction of Shiga toxin-producing Escherichia coli (STEC) as well as antimicrobial resistant (AMR) and non-AMR Salmonella strains on inoculated beef flanks. The RCI technology utilizes a combination of U...

RAPID TETRAZOLIUM DYE REDUCTION ASSAY TO ASSESS THE BACTERICIDAL ACTIVITY OF OYSTER (CRASSOSTREA VIRGINICA) HEMOCYTES AGAINST VIBRIO PARAHAEMOLYTICUS

EPA Science Inventory

An assay was developed to assess the ability of oyster, Crassostrea virginica, hemocytes to kill the human pathogenic bacterium, Vibrio parahaemolyticus (ATCC 17802). Bacterial killing was estimated colorimetrically by the enzymatic reduction of a tetrazolium dye, 3-(4,5-dimethyl...

Optimization, validation, and application of a real-time PCR protocol for quantification of viable bacterial cells in municipal sewage sludge and biosolids using reporter genes and Escherichia coli.

PubMed

van Frankenhuyzen, Jessica K; Trevors, Jack T; Flemming, Cecily A; Lee, Hung; Habash, Marc B

2013-11-01

Biosolids result from treatment of sewage sludge to meet jurisdictional standards, including pathogen reduction. Once government regulations are met, materials can be applied to agricultural lands. Culture-based methods are used to enumerate pathogen indicator microorganisms but may underestimate cell densities, which is partly due to bacteria existing in a viable but non-culturable physiological state. Viable indicators can also be quantified by realtime polymerase chain reaction (qPCR) used with propidium monoazide (PMA), a dye that inhibits amplification of DNA found extracellularly or in dead cells. The objectives of this study were to test an optimized PMA-qPCR method for viable pathogen detection in wastewater solids and to validate it by comparing results to data obtained by conventional plating. Reporter genes from genetically marked Pseudomonas sp. UG14Lr and Agrobacterium tumefaciens 542 cells were spiked into samples of primary sludge, and anaerobically digested and Lystek-treated biosolids as cell-free DNA, dead cells, viable cells, and mixtures of live and dead cells, followed by DNA extraction with and without PMA, and qPCR. The protocol was then used for Escherichia coli quantification in the three matrices, and results compared to plate counts. PMA-qPCR selectively detected viable cells, while inhibiting signals from cell-free DNA and DNA found in membrane-compromised cells. PMA-qPCR detected 0.5-1 log unit more viable E. coli cells in both primary solids and dewatered biosolids than plate counts. No viable E. coli was found in Lystek-treated biosolids. These data suggest PMA-qPCR may more accurately estimate pathogen cell numbers than traditional culture methods.

Microbial risk assessment of drinking water based on hydrodynamic modelling of pathogen concentrations in source water.

PubMed

Sokolova, Ekaterina; Petterson, Susan R; Dienus, Olaf; Nyström, Fredrik; Lindgren, Per-Eric; Pettersson, Thomas J R

2015-09-01

Norovirus contamination of drinking water sources is an important cause of waterborne disease outbreaks. Knowledge on pathogen concentrations in source water is needed to assess the ability of a drinking water treatment plant (DWTP) to provide safe drinking water. However, pathogen enumeration in source water samples is often not sufficient to describe the source water quality. In this study, the norovirus concentrations were characterised at the contamination source, i.e. in sewage discharges. Then, the transport of norovirus within the water source (the river Göta älv in Sweden) under different loading conditions was simulated using a hydrodynamic model. Based on the estimated concentrations in source water, the required reduction of norovirus at the DWTP was calculated using quantitative microbial risk assessment (QMRA). The required reduction was compared with the estimated treatment performance at the DWTP. The average estimated concentration in source water varied between 4.8×10(2) and 7.5×10(3) genome equivalents L(-1); and the average required reduction by treatment was between 7.6 and 8.8 Log10. The treatment performance at the DWTP was estimated to be adequate to deal with all tested loading conditions, but was heavily dependent on chlorine disinfection, with the risk of poor reduction by conventional treatment and slow sand filtration. To our knowledge, this is the first article to employ discharge-based QMRA, combined with hydrodynamic modelling, in the context of drinking water. Copyright © 2015 Elsevier B.V. All rights reserved.

N-Acetylcysteine in Agriculture, a Novel Use for an Old Molecule: Focus on Controlling the Plant–Pathogen Xylella fastidiosa

PubMed Central

Muranaka, Lígia S.; Giorgiano, Thais E.; Takita, Marco A.; Forim, Moacir R.; Silva, Luis F. C.; Coletta-Filho, Helvécio D.; Machado, Marcos A.; de Souza, Alessandra A.

2013-01-01

Xylella fastidiosa is a plant pathogen bacterium that causes diseases in many different crops. In citrus, it causes Citrus Variegated Chlorosis (CVC). The mechanism of pathogenicity of this bacterium is associated with its capacity to colonize and form a biofilm in the xylem vessels of host plants, and there is not yet any method to directly reduce populations of this pathogen in the field. In this study, we investigated the inhibitory effect of N-Acetylcysteine (NAC), a cysteine analogue used mainly to treat human diseases, on X. fastidiosa in different experimental conditions. Concentrations of NAC over 1 mg/mL reduced bacterial adhesion to glass surfaces, biofilm formation and the amount of exopolysaccharides (EPS). The minimal inhibitory concentration of NAC was 6 mg/mL. NAC was supplied to X. fastidiosa-infected plants in hydroponics, fertigation, and adsorbed to organic fertilizer (NAC-Fertilizer). HPLC analysis indicated that plants absorbed NAC at concentrations of 0.48 and 2.4 mg/mL but not at 6 mg/mL. Sweet orange plants with CVC symptoms treated with NAC (0.48 and 2.4 mg/mL) in hydroponics showed clear symptom remission and reduction in bacterial population, as analyzed by quantitative PCR and bacterial isolation. Experiments using fertigation and NAC-Fertilizer were done to simulate a condition closer to that normally is used in the field. For both, significant symptom remission and a reduced bacterial growth rate were observed. Using NAC-Fertilizer the lag for resurgence of symptoms on leaves after interruption of the treatment increased to around eight months. This is the first report of the anti-bacterial effect of NAC against a phytopathogenic bacterium. The results obtained in this work together with the characteristics of this molecule indicate that the use of NAC in agriculture might be a new and sustainable strategy for controlling plant pathogenic bacteria. PMID:24009716

N-acetylcysteine in agriculture, a novel use for an old molecule: focus on controlling the plant-pathogen Xylella fastidiosa.

PubMed

Muranaka, Lígia S; Giorgiano, Thais E; Takita, Marco A; Forim, Moacir R; Silva, Luis F C; Coletta-Filho, Helvécio D; Machado, Marcos A; de Souza, Alessandra A

2013-01-01

Xylella fastidiosa is a plant pathogen bacterium that causes diseases in many different crops. In citrus, it causes Citrus Variegated Chlorosis (CVC). The mechanism of pathogenicity of this bacterium is associated with its capacity to colonize and form a biofilm in the xylem vessels of host plants, and there is not yet any method to directly reduce populations of this pathogen in the field. In this study, we investigated the inhibitory effect of N-Acetylcysteine (NAC), a cysteine analogue used mainly to treat human diseases, on X. fastidiosa in different experimental conditions. Concentrations of NAC over 1 mg/mL reduced bacterial adhesion to glass surfaces, biofilm formation and the amount of exopolysaccharides (EPS). The minimal inhibitory concentration of NAC was 6 mg/mL. NAC was supplied to X. fastidiosa-infected plants in hydroponics, fertigation, and adsorbed to organic fertilizer (NAC-Fertilizer). HPLC analysis indicated that plants absorbed NAC at concentrations of 0.48 and 2.4 mg/mL but not at 6 mg/mL. Sweet orange plants with CVC symptoms treated with NAC (0.48 and 2.4 mg/mL) in hydroponics showed clear symptom remission and reduction in bacterial population, as analyzed by quantitative PCR and bacterial isolation. Experiments using fertigation and NAC-Fertilizer were done to simulate a condition closer to that normally is used in the field. For both, significant symptom remission and a reduced bacterial growth rate were observed. Using NAC-Fertilizer the lag for resurgence of symptoms on leaves after interruption of the treatment increased to around eight months. This is the first report of the anti-bacterial effect of NAC against a phytopathogenic bacterium. The results obtained in this work together with the characteristics of this molecule indicate that the use of NAC in agriculture might be a new and sustainable strategy for controlling plant pathogenic bacteria.

Protection from pulmonary tissue damage associated with infection of cynomolgus macaques by highly pathogenic avian influenza virus (H5N1) by low dose natural human IFN-α administered to the buccal mucosa.

PubMed

Strayer, David R; Carter, William A; Stouch, Bruce C; Stittelaar, Koert J; Thoolen, Robert J M M; Osterhaus, Albert D M E; Mitchell, William M

2014-10-01

Using an established nonhuman primate model for H5N1 highly pathogenic influenza virus infection in humans, we have been able to demonstrate the prophylactic mitigation of the pulmonary damage characteristic of human fatal cases from primary influenza virus pneumonia with a low dose oral formulation of a commercially available parenteral natural human interferon alpha (Alferon N Injection®). At the highest oral dose (62.5IU/kg body weight) used there was a marked reduction in the alveolar inflammatory response with minor evidence of alveolar and interstitial edema in contrast to the hemorrhage and inflammatory response observed in the alveoli of control animals. The mitigation of severe damage to the lower pulmonary airway was observed without a parallel reduction in viral titers. Clinical trial data will be necessary to establish its prophylactic human efficacy for highly pathogenic influenza viruses. Copyright © 2014. Published by Elsevier B.V.

Tick-Pathogen Interactions and Vector Competence: Identification of Molecular Drivers for Tick-Borne Diseases

PubMed Central

de la Fuente, José; Antunes, Sandra; Bonnet, Sarah; Cabezas-Cruz, Alejandro; Domingos, Ana G.; Estrada-Peña, Agustín; Johnson, Nicholas; Kocan, Katherine M.; Mansfield, Karen L.; Nijhof, Ard M.; Papa, Anna; Rudenko, Nataliia; Villar, Margarita; Alberdi, Pilar; Torina, Alessandra; Ayllón, Nieves; Vancova, Marie; Golovchenko, Maryna; Grubhoffer, Libor; Caracappa, Santo; Fooks, Anthony R.; Gortazar, Christian; Rego, Ryan O. M.

2017-01-01

Ticks and the pathogens they transmit constitute a growing burden for human and animal health worldwide. Vector competence is a component of vectorial capacity and depends on genetic determinants affecting the ability of a vector to transmit a pathogen. These determinants affect traits such as tick-host-pathogen and susceptibility to pathogen infection. Therefore, the elucidation of the mechanisms involved in tick-pathogen interactions that affect vector competence is essential for the identification of molecular drivers for tick-borne diseases. In this review, we provide a comprehensive overview of tick-pathogen molecular interactions for bacteria, viruses, and protozoa affecting human and animal health. Additionally, the impact of tick microbiome on these interactions was considered. Results show that different pathogens evolved similar strategies such as manipulation of the immune response to infect vectors and facilitate multiplication and transmission. Furthermore, some of these strategies may be used by pathogens to infect both tick and mammalian hosts. Identification of interactions that promote tick survival, spread, and pathogen transmission provides the opportunity to disrupt these interactions and lead to a reduction in tick burden and the prevalence of tick-borne diseases. Targeting some of the similar mechanisms used by the pathogens for infection and transmission by ticks may assist in development of preventative strategies against multiple tick-borne diseases. PMID:28439499

Effectiveness of cleaning-disinfection wipes and sprays against multidrug-resistant outbreak strains.

PubMed

Kenters, Nikki; Huijskens, Elisabeth G W; de Wit, Sophie C J; van Rosmalen, Joost; Voss, Andreas

2017-08-01

Hospital rooms play an important role in the transmission of several health care-associated pathogens. During the last few years, a number of innovative cleaning-disinfecting products have been brought to market. In this study, commercially available products combining cleaning and disinfection were compared, using 2 different application methods. The aim was to determine which product was most effective in simultaneous cleaning and disinfection of surfaces. Seven cleaning-disinfecting wipes and sprays based on different active ingredients were tested for their efficacy in removal of microbial burden and proteins. Efficacy was tested with known Dutch outbreak strains: vancomycin-resistant enterococci (VRE), Klebsiella pneumoniae OXA-48, or Acinetobacter baumannii. For all bacteria, ready-to-use cleaning-disinfecting products reduced the microbial count with a log 10 reduction >5 with a 5-minute exposure time, with the exception of a spray based on hydrogen peroxide. Omitting the aforementioned hydrogen peroxide spray, there were no significant differences between use of a wipe or spray in bacterial load reduction. Using adenosine triphosphate (ATP) measurements, a significant difference in log 10 relative light units (RLU) reduction between various bacteria (P ≤ .001) was observed. In general, a >5 log 10 reduction of colony forming units (CFU) for tested wipes and sprays was obtained for all tested bacteria strains, with exception of hydrogen peroxide spray and VRE. Although ATP may show a difference between pre- and postcleaning, RLU reduction does not correlate with actual CFU reductions. Copyright © 2017 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Elsevier Inc. All rights reserved.

Disinfection of iceberg lettuce by titanium dioxide-UV photocatalytic reaction.

PubMed

Kim, Youngbong; Choi, Yoonjung; Kim, Soohyun; Park, Jonghyun; Chung, Myongsoo; Song, Kyung Bin; Hwang, Ingyun; Kwon, Kisung; Park, Jiyong

2009-09-01

Securing the physical quality and microbial safety of fresh foods has been a major focus in the food industry. To improve quality and increase the shelf life of fresh produce, disinfection methods have been developed. Titanium dioxide (TiO2) photocatalytic reactions under UV radiation produce hydroxyl radicals that can be used for disinfection of foodborne pathogenic bacteria. We investigated the effects of TiO2-UV photocatalytic disinfection on the shelf life of iceberg lettuce. Counts of natural microflora (total aerobic bacteria, coliforms, psychrotrophic bacteria, and yeasts and molds) and inoculated pathogenic bacteria (Escherichia coli, Listeria monocytogenes, Staphylococcus aureus, and Salmonella Typhimurium) on iceberg lettuce were determined after 20-min treatments with TiO2-UV, UV radiation, a sodium hypochlorite (NaOCl) solution, and tap water. TiO2-UV treatment reduced the number of microorganisms by 1.8 to 2.8 log CFU/g compared with reductions of 0.9 to 1.4 and 0.7 to 1.1 log CFU/g obtained with UV radiation and NaOCl treatments, respectively. Treatment with tap water was used as a control and resulted in no reductions. Counts of microflora for iceberg lettuce at 4 and 25 degrees C were determined during a 9-day period. TiO2-UV treatment resulted in 1.2- and 4.3-log increases in the counts of total aerobic bacteria at 4 and 25 degrees C, respectively, compared with 1.3- to 1.6-log and 4.4- to 4.8-log increases due to UV radiation and NaOCl treatments.

The value of pathogen information in treating clinical mastitis.

PubMed

Cha, Elva; Smith, Rebecca L; Kristensen, Anders R; Hertl, Julia A; Schukken, Ynte H; Tauer, Loren W; Welcome, Frank L; Gröhn, Yrjö T

2016-11-01

The objective of this study was to determine the economic value of obtaining timely and more accurate clinical mastitis (CM) test results for optimal treatment of cows. Typically CM is first identified when the farmer observes recognisable outward signs. Further information of whether the pathogen causing CM is Gram-positive, Gram-negative or other (including no growth) can be determined by using on-farm culture methods. The most detailed level of information for mastitis diagnostics is obtainable by sending milk samples for culture to an external laboratory. Knowing the exact pathogen permits the treatment method to be specifically targeted to the causation pathogen, resulting in less discarded milk. The disadvantages are the additional waiting time to receive test results, which delays treating cows, and the cost of the culture test. Net returns per year (NR) for various levels of information were estimated using a dynamic programming model. The Value of Information (VOI) was then calculated as the difference in NR using a specific level of information as compared to more detailed information on the CM causative agent. The highest VOI was observed where the farmer assumed the pathogen causing CM was the one with the highest incidence in the herd and no pathogen specific CM information was obtained. The VOI of pathogen specific information, compared with non-optimal treatment of Staphylococcus aureus where recurrence and spread occurred due to lack of treatment efficacy, was $20.43 when the same incorrect treatment was applied to recurrent cases, and $30.52 when recurrent cases were assumed to be the next highest incidence pathogen and treated accordingly. This indicates that negative consequences associated with choosing the wrong CM treatment can make additional information cost-effective if pathogen identification is assessed at the generic information level and if the pathogen can spread to other cows if not treated appropriately.

Fast Disinfection of Escherichia coli Bacteria Using Carbon Nanotubes Interaction with Microwave Radiation

PubMed Central

Al-Hakami, Samer M.; Khalil, Amjad B.; Laoui, Tahar; Atieh, Muataz Ali

2013-01-01

Water disinfection has attracted the attention of scientists worldwide due to water scarcity. The most significant challenges are determining how to achieve proper disinfection without producing harmful byproducts obtained usually using conventional chemical disinfectants and developing new point-of-use methods for the removal and inactivation of waterborne pathogens. The removal of contaminants and reuse of the treated water would provide significant reductions in cost, time, liabilities, and labour to the industry and result in improved environmental stewardship. The present study demonstrates a new approach for the removal of Escherichia coli (E. coli) from water using as-produced and modified/functionalized carbon nanotubes (CNTs) with 1-octadecanol groups (C18) under the effect of microwave irradiation. Scanning/transmission electron microscopy, thermogravimetric analysis, and FTIR spectroscopy were used to characterise the morphological/structural and thermal properties of CNTs. The 1-octadecanol (C18) functional group was attached to the surface of CNTs via Fischer esterification. The produced CNTs were tested for their efficiency in destroying the pathogenic bacteria (E. coli) in water with and without the effect of microwave radiation. A low removal rate (3–5%) of (E. coli) bacteria was obtained when CNTs alone were used, indicating that CNTs did not cause bacterial cellular death. When combined with microwave radiation, the unmodified CNTs were able to remove up to 98% of bacteria from water, while a higher removal of bacteria (up to 100%) was achieved when CNTs-C18 was used under the same conditions. PMID:23606820

The efficacy of Mentha arvensis L. and M. piperita L. essential oils in reducing pathogenic bacteria and maintaining quality characteristics in cashew, guava, mango, and pineapple juices.

PubMed

de Sousa Guedes, Jossana Pereira; da Costa Medeiros, José Alberto; de Souza E Silva, Richard Sidney; de Sousa, Janaína Maria Batista; da Conceição, Maria Lúcia; de Souza, Evandro Leite

2016-12-05

This study evaluated the ability of the essential oil from Mentha arvensis L. (MAEO) and M. piperita L. (MPEO) to induce ≥5-log reductions in counts (CFU/mL) of E. coli, L. monocytogenes, and Salmonella enterica serovar Enteritidis in Brain-Heart Infusion broth (BHIB) and cashew, guava, mango, and pineapple juices during refrigerated storage (4±0.5°C). The effects of the incorporation of these essential oils on some physicochemical and sensory parameters of juices were also evaluated. The incorporation of 5, 2.5, 1.25, or 0.625μL/mL of MAEO in BHIB caused a ≥5-log reduction in counts of E. coli and Salmonella Enteritidis after 24h of storage; but only 5μL/mL was able to cause the same reduction in counts of L.monocytogenes. The incorporation of 10μL/mL of MPEO in BHIB caused a ≥5-log reduction in counts of E. coli, Salmonella Enteritidis, and L. monocytogenes after 24h of storage; smaller reductions were observed in BHIB containing 5, 2.5, and 1.25μL/mL of MPEO. Similar reductions were observed when the MAEO or MPEO was incorporated at the same concentrations in mango juice. The incorporation of MAEO or MPEO at all tested concentrations in cashew, guava, and pineapple juices resulted in a ≥5-log reduction in pathogen counts within 1h. The incorporation of MAEO and MPEO (0.625 and 1.25μL/mL, respectively) in fruit juices did not induce alterations in °Brix, pH, and acidity, but negatively affected the taste, aftertaste, and overall acceptance. The use of MAEO or MPEO at low concentrations could constitute an interesting tool to achieve the required 5-log reduction of pathogenic bacteria in cashew, guava, mango, and pineapple fruit juices. However, new methods combining the use of MAEO or MPEO with other technologies are necessary to reduce their negative impacts on specific sensory properties of these juices. Copyright © 2016 Elsevier B.V. All rights reserved.

40 CFR 503.17 - Recordkeeping.

Code of Federal Regulations, 2011 CFR

2011-07-01

... in § 503.32(a) and the vector attraction reduction requirement in [insert one of the vector attraction reduction requirements in § 503.33(b)(1) through § 503.33(b)(8)] was prepared under my direction... pathogen requirements in § 503.32(a) are met. (iv) A description of how one of the vector attraction...

Inducible expression of Bs2 R gene from Capsicum chacoense in sweet orange (Citrus sinensis L. Osbeck) confers enhanced resistance to citrus canker disease.

PubMed

Sendín, Lorena Noelia; Orce, Ingrid Georgina; Gómez, Rocío Liliana; Enrique, Ramón; Grellet Bournonville, Carlos Froilán; Noguera, Aldo Sergio; Vojnov, Adrián Alberto; Marano, María Rosa; Castagnaro, Atilio Pedro; Filippone, María Paula

2017-04-01

Transgenic expression of the pepper Bs2 gene confers resistance to Xanthomonas campestris pv. vesicatoria (Xcv) pathogenic strains which contain the avrBs2 avirulence gene in susceptible pepper and tomato varieties. The avrBs2 gene is highly conserved among members of the Xanthomonas genus, and the avrBs2 of Xcv shares 96% homology with the avrBs2 of Xanthomonas citri subsp. citri (Xcc), the causal agent of citrus canker disease. A previous study showed that the transient expression of pepper Bs2 in lemon leaves reduced canker formation and induced plant defence mechanisms. In this work, the effect of the stable expression of Bs2 gene on citrus canker resistance was evaluated in transgenic plants of Citrus sinensis cv. Pineapple. Interestingly, Agrobacterium-mediated transformation of epicotyls was unsuccessful when a constitutive promoter (2× CaMV 35S) was used in the plasmid construction, but seven transgenic lines were obtained with a genetic construction harbouring Bs2 under the control of a pathogen-inducible promoter, from glutathione S-transferase gene from potato. A reduction of disease symptoms of up to 70% was observed in transgenic lines expressing Bs2 with respect to non-transformed control plants. This reduction was directly dependent on the Xcc avrBs2 gene since no effect was observed when a mutant strain of Xcc with a disruption in avrBs2 gene was used for inoculations. Additionally, a canker symptom reduction was correlated with levels of the Bs2 expression in transgenic plants, as assessed by real-time qPCR, and accompanied by the production of reactive oxygen species. These results indicate that the pepper Bs2 resistance gene is also functional in a family other than the Solanaceae, and could be considered for canker control.

Tyr120Asp mutation alters domain flexibility and dynamics of MeCP2 DNA binding domain leading to impaired DNA interaction: Atomistic characterization of a Rett syndrome causing mutation.

PubMed

D'Annessa, Ilda; Gandaglia, Anna; Brivio, Elena; Stefanelli, Gilda; Frasca, Angelisa; Landsberger, Nicoletta; Di Marino, Daniele

2018-05-01

Mutations in the X-linked MECP2 gene represent the main origin of Rett syndrome, causing a profound intellectual disability in females. MeCP2 is an epigenetic transcriptional regulator containing two main functional domains: a methyl-CpG binding domain (MBD) and a transcription repression domain (TRD). Over 600 pathogenic mutations were reported to affect the whole protein; almost half of missense mutations affect the MBD. Understanding the impact of these mutations on the MBD structure and interaction with DNA will foster the comprehension of their pathogenicity and possibly genotype/phenotype correlation studies. Herein, we use molecular dynamics simulations to obtain a detailed view of the dynamics of WT and mutated MBD in the presence and absence of DNA. The pathogenic mutation Y120D is used as paradigm for our studies. Further, since the Y120 residue was previously found to be a phosphorylation site, we characterize the dynamic profile of the MBD also in the presence of Y120 phosphorylation (pY120). We found that addition of a phosphate group to Y120 or mutation in aspartic acid affect domain mobility that samples an alternative conformational space with respect to the WT, leading to impaired ability to interact with DNA. Experimental assays showing a significant reduction in the binding affinity between the mutated MBD and the DNA confirmed our predictions. Copyright © 2018 Elsevier B.V. All rights reserved.

Effect of Larval Competition on Extrinsic Incubation Period and Vectorial Capacity of Aedes albopictus for Dengue Virus.

PubMed

Bara, Jeffrey; Rapti, Zoi; Cáceres, Carla E; Muturi, Ephantus J

2015-01-01

Despite the growing awareness that larval competition can influence adult mosquito life history traits including susceptibility to pathogens, the net effect of larval competition on human risk of exposure to mosquito-borne pathogens remains poorly understood. We examined how intraspecific larval competition affects dengue-2 virus (DENV-2) extrinsic incubation period and vectorial capacity of its natural vector Aedes albopictus. Adult Ae. albopictus from low and high-larval density conditions were orally challenged with DENV-2 and then assayed for virus infection and dissemination rates following a 6, 9, or 12-day incubation period using real-time quantitative reverse transcription PCR. We then modeled the effect of larval competition on vectorial capacity using parameter estimates obtained from peer-reviewed field and laboratory studies. Larval competition resulted in significantly longer development times, lower emergence rates, and smaller adults, but did not significantly affect the extrinsic incubation period of DENV-2 in Ae. albopictus. Our vectorial capacity models suggest that the effect of larval competition on adult mosquito longevity likely has a greater influence on vectorial capacity relative to any competition-induced changes in vector competence. Furthermore, we found that large increases in the viral dissemination rate may be necessary to compensate for small competition-induced reductions in daily survivorship. Our results indicate that mosquito populations that experience stress from larval competition are likely to have a reduced vectorial capacity, even when susceptibility to pathogens is enhanced.

Effect of Larval Competition on Extrinsic Incubation Period and Vectorial Capacity of Aedes albopictus for Dengue Virus

PubMed Central

Bara, Jeffrey; Rapti, Zoi; Cáceres, Carla E.; Muturi, Ephantus J.

2015-01-01

Despite the growing awareness that larval competition can influence adult mosquito life history traits including susceptibility to pathogens, the net effect of larval competition on human risk of exposure to mosquito-borne pathogens remains poorly understood. We examined how intraspecific larval competition affects dengue-2 virus (DENV-2) extrinsic incubation period and vectorial capacity of its natural vector Aedes albopictus. Adult Ae. albopictus from low and high-larval density conditions were orally challenged with DENV-2 and then assayed for virus infection and dissemination rates following a 6, 9, or 12-day incubation period using real-time quantitative reverse transcription PCR. We then modeled the effect of larval competition on vectorial capacity using parameter estimates obtained from peer-reviewed field and laboratory studies. Larval competition resulted in significantly longer development times, lower emergence rates, and smaller adults, but did not significantly affect the extrinsic incubation period of DENV-2 in Ae. albopictus. Our vectorial capacity models suggest that the effect of larval competition on adult mosquito longevity likely has a greater influence on vectorial capacity relative to any competition-induced changes in vector competence. Furthermore, we found that large increases in the viral dissemination rate may be necessary to compensate for small competition-induced reductions in daily survivorship. Our results indicate that mosquito populations that experience stress from larval competition are likely to have a reduced vectorial capacity, even when susceptibility to pathogens is enhanced. PMID:25951173

Assessment of the effect of a Salmonella enterica ser. Typhimurium culture supernatant on the single-cell lag time of foodborne pathogens.

PubMed

Blana, Vasiliki A; Lianou, Alexandra; Nychas, George-John E

2015-12-23

The objective of this study was the in vitro evaluation of the effect of a cell-free microbial supernatant, produced by a luxS-positive Salmonella enterica ser. Typhimurium strain, on the single-cell growth kinetic behavior of two strains of S. enterica (serotypes Enteritidis and Typhimurium) and a methicillin-resistant Staphylococcus aureus strain. The single-cell lag time (λ) of the pathogens was estimated in the absence and presence (20% v/v) of microbial supernatant based on optical density measurements. As demonstrated by the obtained results, the tested microbial supernatant had a strain-specific effect on the single-cell λ and its variability. Although the mean λ values were similar in the absence and presence of microbial supernatant in the case of Salmonella Enteritidis, a significant (P ≤ 0.05) reduction and increase in the mean value of this parameter in the presence of microbial supernatant were observed for Salmonella Typhimurium and St. aureus, respectively. With regard to the effect of the tested microbial supernatant on the single-cell variability of λ, similar λ distributions were obtained in its absence and presence for S. Enteritidis, while considerable differences were noted for the other two tested organisms; the coefficient of variation of λ in the absence and presence of microbial supernatant was 41.6 and 69.8% for S. Typhimurium, respectively, with the corresponding values for St. aureus being 74.0 and 56.9%. As demonstrated by the results of bioassays, the tested microbial supernatant exhibited autoinducer-2 activity, indicating a potential association of such quorum sensing compounds with the observed effects. Although preliminary in nature, the collected data provide a good basis for future research on the role of quorum sensing in the single-cell growth behavior of foodborne pathogens.

Pathogen Inactivated Plasma Concentrated: Preparation and Uses

DTIC Science & Technology

2004-09-01

REPORT DATE 01 SEP 2004 2 . REPORT TYPE N/A 3. DATES COVERED - 4. TITLE AND SUBTITLE Pathogen Inactivated Plasma Concentrated: Preparation...Concentrated: Preparation and Uses 22 - 2 RTO-MP-HFM-109 Results: Both UVC and ozone yielded a PPV logarithmic reduction factor (LRF) of 6, for a...technology to be marketed; the industry name is Plas+SD [ 2 ]. This process functions by attacking the lipid sheathes that surround enveloped viruses

9 CFR 113.36 - Detection of pathogens by the chicken inoculation test.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Detection of pathogens by the chicken... REQUIREMENTS Standard Procedures § 113.36 Detection of pathogens by the chicken inoculation test. The test for...,000 doses. (b) At least 25 healthy susceptible young chickens, properly identified and obtained from...

Use of cationic polymers to reduce pathogen levels during dairy manure separation.

PubMed

Liu, Zong; Carroll, Zachary S; Long, Sharon C; Gunasekaran, Sundaram; Runge, Troy

2016-01-15

Various separation technologies are used to deal with the enormous amounts of animal waste that large livestock operations generate. When the recycled waste stream is land applied, it is essential to lower the pathogen load to safeguard the health of livestock and humans. We investigated whether cationic polymers, used as a flocculent in the solid/liquid separation process, could reduce the pathogen indicator load in the animal waste stream. The effects of low charge density cationic polyacrylamide (CPAM) and high charge density cationic polydicyandiamide (PDCD) were investigated. Results demonstrated that CPAM was more effective than PDCD for manure coagulation and flocculation, while PDCD was more effective than CPAM in reducing the pathogen indicator loads. However, their combined use, CPAM followed by PDCD, resulted in both improved solids separation and pathogen indicator reduction. Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.

Potential benefits of chewing gum for the delivery of oral therapeutics and its possible role in oral healthcare.

PubMed

Wessel, Stefan W; van der Mei, Henny C; Maitra, Amarnath; Dodds, Michael W J; Busscher, Henk J

2016-10-01

Over the years, chewing gum has developed from a candy towards an oral health-promoting nutraceutical. This review summarizes evidence for the oral health benefits of chewing gum, emphasizing identification of active ingredients in gum that facilitate prevention and removal of oral biofilm. Chewing of sugar-free gum yields oral health benefits that include clearance of food debris, reduction in oral dryness, increase of biofilm pH and remineralization of enamel. These basic effects of chewing gum are attributed to increased mastication and salivation. Active ingredients incorporated in chewing gums aim to expand these effects to inhibition of extrinsic tooth stain and calculus formation, enhanced enamel remineralization, reduction of the numbers of bacteria in saliva and amount of oral biofilm, neutralization of biofilm pH, and reduction of volatile sulfur compounds. Evidence for oral-health benefits of chewing gum additives is hard to obtain due to their relatively low concentrations and rapid wash-out. Clinical effects of gum additives are overshadowed by effects of increased mastication and salivation due to the chewing of gum and require daily chewing of gum for prolonged periods of time. Future studies on active ingredients should focus on specifically targeting pathogenic bacteria, whilst leaving the healthy microbiome unaffected.

Modeling the survival of Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella Typhimurium during fermentation, drying, and storage of soudjouk-style fermented sausage.

PubMed

Hwang, Cheng-An; Porto-Fett, Anna C S; Juneja, Vijay K; Ingham, Steven C; Ingham, Barbara H; Luchansky, John B

2009-02-28

This study quantified and modeled the survival of Escherichia coli O157:H7, Listeria monocytogenes and Salmonella Typhimurium in soudjouk-style fermented sausage during fermentation, drying, and storage. Batter prepared from ground beef (20% fat), seasonings, starter culture, and dextrose was separately inoculated with a multi-strain mixture of each pathogen to an initial inoculum of ca. 6.5 log(10) CFU/g in the batter. The sausages were subsequently fermented at 24 degrees C with a relative humidity (RH) of 90% to 95% for 3 to 5 days to ca. pH 5.2, pH 4.9 or pH 4.6, then dried at 22 degrees C to a(w) 0.92, a(w) 0.89, or a(w) 0.86, respectively, and then stored at 4, 21, or 30 degrees C for up to 60 days. Lethality of the three pathogens was modeled as a function of pH, a(w) and/or storage temperature. During fermentation to pH 5.2 to pH 4.6, cell reductions ranged from 0 to 0.9 log(10) CFU/g for E. coli O157:H7, 0.1 to 0.5 log(10) CFU/g for L. monocytogenes, and 0 to 2.2 log(10) CFU/g for S. Typhimurium. Subsequent drying of sausages of pH 5.2 to pH 4.6 at 22 degrees C with 80% to 85% RH for 3 to 7 days to a(w) of 0.92 to a(w) 0.86 resulted in additional reductions that ranged from 0 to 3.5 log(10) CFU/g for E. coli O157:H7, 0 to 0.4 log(10) CFU/g for L. monocytogenes, and 0.3 to 2.4 log(10) CFU/g for S. Typhimurium. During storage at 4, 21, or 30 degrees C the reduction rates of the three pathogens were generally higher (p<0.05) in sausages with lower pH and lower a(w) that were stored at higher temperatures. Polynomial equations were developed to describe the inactivation of the three pathogens during fermentation, drying, and storage. The applicability of the resulting models for fermented sausage was evaluated by comparing model predictions with published data. Pathogen reductions estimated by the models for E. coli O157:H7 and S. Typhimurium were comparable to 67% and 73% of published data, respectively. Due to limited published data for L. monocytogenes, the models for L. monocytogenes would need additional validations. Results of pathogen reductions from this study may be used as a reference to assist manufacturers of soudjouk-style sausages to adopt manufacturing processes that meet the regulatory requirements. The resulting models may also be used for estimating the survival of E. coli O157:H7 and S. Typhimurium in other similar fermented sausage during fermentation and storage.

Application of Proteomics for the Investigation of the Effect of Initial pH on Pathogenic Mechanisms of Fusarium proliferatum on Banana Fruit.

PubMed

Li, Taotao; Wu, Qixian; Wang, Yong; John, Afiya; Qu, Hongxia; Gong, Liang; Duan, Xuewu; Zhu, Hong; Yun, Ze; Jiang, Yueming

2017-01-01

Fusarium proliferatum is an important pathogen and causes a great economic loss to fruit industry. Environmental pH-value plays a regulatory role in fungi pathogenicity, however, the mechanism needs further exploration. In this study, F. proliferatum was cultured under two initial pH conditions of 5 and 10. No obvious difference was observed in the growth rate of F. proliferatum between two pH-values. F. proliferatum cultured under both pH conditions infected banana fruit successfully, and smaller lesion diameter was presented on banana fruit inoculated with pH 10-cultured fungi. Proteomic approach based on two-dimensional electrophoresis (2-DE) was used to investigate the changes in secretome of this fungus between pH 5 and 10. A total of 39 differential spots were identified using matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry (MALDI-TOF/TOF-MS) and liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS). Compared to pH 5 condition, proteins related to cell wall degrading enzymes (CWDEs) and proteolysis were significantly down-regulated at pH 10, while proteins related to oxidation-reduction process and transport were significantly up-regulated under pH 10 condition. Our results suggested that the downregulation of CWDEs and other virulence proteins in the pH 10-cultured F. proliferatum severely decreased its pathogenicity, compared to pH 5-cultured fungi. However, the alkaline environment did not cause a complete loss of the pathogenic ability of F. proliferatum , probably due to the upregulation of the oxidation-reduction related proteins at pH 10, which may partially compensate its pathogenic ability.

Neem (Azadirachta indica A. Juss) Oil to Tackle Enteropathogenic Escherichia coli

PubMed Central

Del Serrone, Paola; Nicoletti, Marcello

2015-01-01

Neem (Azadirachta indica A. Juss) oil (NO) was assayed against forty-eight isolates of Escherichia coli by standardised disc diffusion test and microdilution test. By molecular biology characterization, fourteen isolates resulted in diarrheagenic E. coli with sixteen primer pairs that specifically amplify unique sequences of virulence genes and of 16S rRNA. The NO showed biological activity against all isolates. The bacterial growth inhibition zone by disc diffusion method (100 µL NO) ranged between 9.50 ± 0.70 and 30.00 ± 1.00 mm. The antibacterial activity was furthermore determined at lower NO concentrations (1 : 10–1 : 10,000). The percent of growth reduction ranged between 23.71 ± 1.00 and 99.70 ± 1.53. The highest bacterial growth reduction was 1 : 10 NO concentration with 50 µL of bacterial suspension (ca. 1 × 106 CFU/mL). There is significant difference between the antibacterial activities against pathogenic and nonpathogenic E. coli, as well as NO and ciprofloxacin activities. Viable cells after the different NO concentration treatments were checked by molecular biology assay using PMA dye. On the basis of the obtained results, NO counteracts E. coli and also influences the virulence of E. coli viable cells after NO treatment. The NO metabolomic composition was obtained using fingerprint HPTLC. PMID:26064900

Neem (Azadirachta indica A. Juss) Oil to Tackle Enteropathogenic Escherichia coli.

PubMed

Del Serrone, Paola; Toniolo, Chiara; Nicoletti, Marcello

2015-01-01

Neem (Azadirachta indica A. Juss) oil (NO) was assayed against forty-eight isolates of Escherichia coli by standardised disc diffusion test and microdilution test. By molecular biology characterization, fourteen isolates resulted in diarrheagenic E. coli with sixteen primer pairs that specifically amplify unique sequences of virulence genes and of 16S rRNA. The NO showed biological activity against all isolates. The bacterial growth inhibition zone by disc diffusion method (100 µL NO) ranged between 9.50 ± 0.70 and 30.00 ± 1.00 mm. The antibacterial activity was furthermore determined at lower NO concentrations (1 : 10-1 : 10,000). The percent of growth reduction ranged between 23.71 ± 1.00 and 99.70 ± 1.53. The highest bacterial growth reduction was 1 : 10 NO concentration with 50 µL of bacterial suspension (ca. 1 × 10(6) CFU/mL). There is significant difference between the antibacterial activities against pathogenic and nonpathogenic E. coli, as well as NO and ciprofloxacin activities. Viable cells after the different NO concentration treatments were checked by molecular biology assay using PMA dye. On the basis of the obtained results, NO counteracts E. coli and also influences the virulence of E. coli viable cells after NO treatment. The NO metabolomic composition was obtained using fingerprint HPTLC.

Spectroscopy investigation on chemo-catalytic, free radical scavenging and bactericidal properties of biogenic silver nanoparticles synthesized using Salicornia brachiata aqueous extract

NASA Astrophysics Data System (ADS)

Seralathan, Janani; Stevenson, Priscilla; Subramaniam, Shankar; Raghavan, Rachana; Pemaiah, Brindha; Sivasubramanian, Aravind; Veerappan, Anbazhagan

2014-01-01

Nanosized silver have been widely used in many applications, such as catalysis, photonics, sensors, medicine etc. Thus, there is an increasing need to develop high-yield, low cost, non-toxic and eco-friendly procedures for the synthesis of nanoparticles. Herein, we report an efficient, green synthesis of silver nanoparticles utilizing the aqueous extract of Salicornia brachiata, a tropical plant of the Chenopodiaceae family. Silver nanoparticles have been characterized by ultraviolet-visible spectroscopy, scanning electron microscopy and transmission electron microscopy. The morphology of the particles formed consists of highly diversified shapes like spherical, rod-like, prism, triangular, pentagonal and hexagonal pattern. However, addition of sodium hydroxide to the extract produces mostly spherical particles. The stable nanoparticles obtained using this green method show remarkable catalytic activity in the reduction of 4-nitro phenol to 4-amino phenol. The reduction catalyzed by silver nanoparticles followed the first-order kinetics, with a rate constant of, 0.6 × 10-2 s-1. The bactericidal activity of the synthesized silver nanoparticles against the pathogenic bacteria, Staphylococcus aureus, Staphylococcus aureus E, Bacillus subtilis and Escherichia coli, was also explored using REMA. The obtained results showed that the minimum inhibitory concentration required to induce bactericidal effect is lower than the control antibiotic, ciprofloxacin. In addition to these, the biogenic synthesized nanoparticles also exhibited excellent free radical scavenging activity.

Whole-leaf sanitizing wash improves chlorine efficacy for microbial reduction and prevents pathogen cross contamination during fresh-cut lettuce processing

USDA-ARS?s Scientific Manuscript database

Currently, nearly all fresh-cut lettuce processing facilities in the United States use chlorinated water or other sanitizer solutions for microbial reduction after lettuce is cut. It is believed that freshly cut lettuce releases significant amounts of organic matters that negatively impact the effec...

The transcriptome of Spodoptera exigua larvae exposed to different types of microbes.

PubMed

Pascual, Laura; Jakubowska, Agata K; Blanca, Jose M; Cañizares, Joaquin; Ferré, Juan; Gloeckner, Gernot; Vogel, Heiko; Herrero, Salvador

2012-08-01

We have obtained and characterized the transcriptome of Spodoptera exigua larvae with special emphasis on pathogen-induced genes. In order to obtain a highly representative transcriptome, we have pooled RNA from diverse insect colonies, conditions and tissues. Sequenced cDNA included samples from 3 geographically different colonies. Enrichment of RNA from pathogen-related genes was accomplished by exposing larvae to different pathogenic and non-pathogenic microbial agents such as the bacteria Bacillus thuringiensis, Micrococcus luteus, and Escherichia coli, the yeast Saccharomyces cerevisiae, and the S. exigua nucleopolyhedrovirus (SeMNPV). In addition, to avoid the loss of tissue-specific genes we included cDNA from the midgut, fat body, hemocytes and integument derived from pathogen exposed insects. RNA obtained from the different types of samples was pooled, normalized and sequenced. Analysis of the sequences obtained using the Roche 454 FLX and Sanger methods has allowed the generation of the largest public set of ESTs from S. exigua, including a large group of immune genes, and the identification of an important number of SSR (simple sequence repeats) and SNVs (single nucleotide variants: SNPs and INDELs) with potential use as genetic markers. Moreover, data mining has allowed the discovery of novel RNA viruses with potential influence in the insect population dynamics and the larval interactions with the microbial pesticides that are currently in use for the biological control of this pest. Copyright © 2012 Elsevier Ltd. All rights reserved.

Potential burden of antibiotic resistance on surgery and cancer chemotherapy antibiotic prophylaxis in the USA: a literature review and modelling study.

PubMed

Teillant, Aude; Gandra, Sumanth; Barter, Devra; Morgan, Daniel J; Laxminarayan, Ramanan

2015-12-01

The declining efficacy of existing antibiotics potentially jeopardises outcomes in patients undergoing medical procedures. We investigated the potential consequences of increases in antibiotic resistance on the ten most common surgical procedures and immunosuppressing cancer chemotherapies that rely on antibiotic prophylaxis in the USA. We searched the published scientific literature and identified meta-analyses and reviews of randomised controlled trials or quasi-randomised controlled trials (allocation done on the basis of a pseudo-random sequence-eg, odd/even hospital number or date of birth, alternation) to estimate the efficacy of antibiotic prophylaxis in preventing infections and infection-related deaths after surgical procedures and immunosuppressing cancer chemotherapy. We varied the identified effect sizes under different scenarios of reduction in the efficacy of antibiotic prophylaxis (10%, 30%, 70%, and 100% reductions) and estimated the additional number of infections and infection-related deaths per year in the USA for each scenario. We estimated the percentage of pathogens causing infections after these procedures that are resistant to standard prophylactic antibiotics in the USA. We estimate that between 38·7% and 50·9% of pathogens causing surgical site infections and 26·8% of pathogens causing infections after chemotherapy are resistant to standard prophylactic antibiotics in the USA. A 30% reduction in the efficacy of antibiotic prophylaxis for these procedures would result in 120,000 additional surgical site infections and infections after chemotherapy per year in the USA (ranging from 40,000 for a 10% reduction in efficacy to 280,000 for a 70% reduction in efficacy), and 6300 infection-related deaths (range: 2100 for a 10% reduction in efficacy, to 15,000 for a 70% reduction). We estimated that every year, 13,120 infections (42%) after prostate biopsy are attributable to resistance to fluoroquinolones in the USA. Increasing antibiotic resistance potentially threatens the safety and efficacy of surgical procedures and immunosuppressing chemotherapy. More data are needed to establish how antibiotic prophylaxis recommendations should be modified in the context of increasing rates of resistance. DRIVE-AB Consortium. Copyright © 2015 Elsevier Ltd. All rights reserved.

Staphylococcus saprophyticus surface-associated protein (Ssp) is associated with lifespan reduction in Caenorhabditis elegans.

PubMed

Szabados, Florian; Mohner, Amelie; Kleine, Britta; Gatermann, Sören G

2013-10-01

Staphylococcal lipases have been proposed as pathogenicity factors. In Staphylococcus saprophyticus the surface-associated protein (Ssp) has been previously characterized as a cell wall-associated true lipase. A S. saprophyticus Δssp::ermB mutant has been described as less virulent in an in vivo model of urinary tract infection compared with its wild-type. This is the first report showing that S. saprophyticus induced a lifespan reduction in Caenorhabditis elegans similar to that of S. aureus RN4220. In two S. saprophyticus Δssp::ermB mutants lifespan reduction in C. elegans was partly abolished. In order to attribute virulence to the lipase activity itself and distinguish this phenomenon from the presence of the Ssp-protein, the conserved active site of the lipase was modified by site-directed ligase-independent mutagenesis and lipase activity-deficient mutants were constructed. These results indicate that the Ssp is associated with pathogenicity in C. elegans and one could speculate that the lipase activity itself is responsible for this virulence.

Staphylococcus saprophyticus surface-associated protein (Ssp) is associated with lifespan reduction in Caenorhabditis elegans

PubMed Central

Szabados, Florian; Mohner, Amelie; Kleine, Britta; Gatermann, Sören G

2013-01-01

Staphylococcal lipases have been proposed as pathogenicity factors. In Staphylococcus saprophyticus the surface-associated protein (Ssp) has been previously characterized as a cell wall-associated true lipase. A S. saprophyticus Δssp::ermB mutant has been described as less virulent in an in vivo model of urinary tract infection compared with its wild-type. This is the first report showing that S. saprophyticus induced a lifespan reduction in Caenorhabditis elegans similar to that of S. aureus RN4220. In two S. saprophyticus Δssp::ermB mutants lifespan reduction in C. elegans was partly abolished. In order to attribute virulence to the lipase activity itself and distinguish this phenomenon from the presence of the Ssp-protein, the conserved active site of the lipase was modified by site-directed ligase-independent mutagenesis and lipase activity-deficient mutants were constructed. These results indicate that the Ssp is associated with pathogenicity in C. elegans and one could speculate that the lipase activity itself is responsible for this virulence. PMID:23959029

A theoretical framework for biological control of soil-borne plant pathogens: Identifying effective strategies.

PubMed

Cunniffe, Nik J; Gilligan, Christopher A

2011-06-07

We develop and analyse a flexible compartmental model of the interaction between a plant host, a soil-borne pathogen and a microbial antagonist, for use in optimising biological control. By extracting invasion and persistence thresholds of host, pathogen and biological control agent, performing an equilibrium analysis, and numerical investigation of sensitivity to parameters and initial conditions, we determine criteria for successful biological control. We identify conditions for biological control (i) to prevent a pathogen entering a system, (ii) to eradicate a pathogen that is already present and, if that is not possible, (iii) to reduce the density of the pathogen. Control depends upon the epidemiology of the pathogen and how efficiently the antagonist can colonise particular habitats (i.e. healthy tissue, infected tissue and/or soil-borne inoculum). A sharp transition between totally effective control (i.e. eradication of the pathogen) and totally ineffective control can follow slight changes in biologically interpretable parameters or to the initial amounts of pathogen and biological control agent present. Effective biological control requires careful matching of antagonists to pathosystems. For preventative/eradicative control, antagonists must colonise susceptible hosts. However, for reduction in disease prevalence, the range of habitat is less important than the antagonist's bulking-up efficiency. Copyright © 2011 Elsevier Ltd. All rights reserved.

Evolution of pathogen virulence across space during an epidemic

USGS Publications Warehouse

Osnas, Erik; Hurtado, Paul J.; Dobson, Andrew P.

2015-01-01

We explore pathogen virulence evolution during the spatial expansion of an infectious disease epidemic in the presence of a novel host movement trade-off, using a simple, spatially explicit mathematical model. This work is motivated by empirical observations of the Mycoplasma gallisepticum invasion into North American house finch (Haemorhous mexicanus) populations; however, our results likely have important applications to other emerging infectious diseases in mobile hosts. We assume that infection reduces host movement and survival and that across pathogen strains the severity of these reductions increases with pathogen infectiousness. Assuming these trade-offs between pathogen virulence (host mortality), pathogen transmission, and host movement, we find that pathogen virulence levels near the epidemic front (that maximize wave speed) are lower than those that have a short-term growth rate advantage or that ultimately prevail (i.e., are evolutionarily stable) near the epicenter and where infection becomes endemic (i.e., that maximize the pathogen basic reproductive ratio). We predict that, under these trade-offs, less virulent pathogen strains will dominate the periphery of an epidemic and that more virulent strains will increase in frequency after invasion where disease is endemic. These results have important implications for observing and interpreting spatiotemporal epidemic data and may help explain transient virulence dynamics of emerging infectious diseases.

Pathogen reduction by ultraviolet C light effectively inactivates human white blood cells in platelet products.

PubMed

Pohler, Petra; Müller, Meike; Winkler, Carla; Schaudien, Dirk; Sewald, Katherina; Müller, Thomas H; Seltsam, Axel

2015-02-01

Residual white blood cells (WBCs) in cellular blood components induce a variety of adverse immune events, including nonhemolytic febrile transfusion reactions, alloimmunization to HLA antigens, and transfusion-associated graft-versus-host disease (TA-GVHD). Pathogen reduction (PR) methods such as the ultraviolet C (UVC) light-based THERAFLEX UV-Platelets system were developed to reduce the risk of transfusion-transmitted infection. As UVC light targets nucleic acids, it interferes with the replication of both pathogens and WBCs. This preclinical study aimed to evaluate the ability of UVC light to inactivate contaminating WBCs in platelet concentrates (PCs). The in vitro and in vivo function of WBCs from UVC-treated PCs was compared to that of WBCs from gamma-irradiated and untreated PCs by measuring cell viability, proliferation, cytokine secretion, antigen presentation in vitro, and xenogeneic GVHD responses in a humanized mouse model. UVC light was at least as effective as gamma irradiation in preventing GVHD in the mouse model. It was more effective in suppressing T-cell proliferation (>5-log reduction in the limiting dilution assay), cytokine secretion, and antigen presentation than gamma irradiation. The THERAFLEX UV-Platelets (MacoPharma) PR system can substitute gamma irradiation for TA-GVHD prophylaxis in platelet (PLT) transfusion. Moreover, UVC treatment achieves suppression of antigen presentation and inhibition of cytokine accumulation during storage of PCs, which has potential benefits for transfusion recipients. © 2014 AABB.

Nitrite addition to acidified sludge significantly improves digestibility, toxic metal removal, dewaterability and pathogen reduction

NASA Astrophysics Data System (ADS)

Du, Fangzhou; Keller, Jürg; Yuan, Zhiguo; Batstone, Damien J.; Freguia, Stefano; Pikaar, Ilje

2016-12-01

Sludge management is a major issue for water utilities globally. Poor digestibility and dewaterability are the main factors determining the cost for sludge management, whereas pathogen and toxic metal concentrations limit beneficial reuse. In this study, the effects of low level nitrite addition to acidified sludge to simultaneously enhance digestibility, toxic metal removal, dewaterability and pathogen reduction were investigated. Waste activated sludge (WAS) from a full-scale waste water treatment plant was treated at pH 2 with 10 mg NO2--N/L for 5 h. Biochemical methane potential tests showed an increase in the methane production of 28%, corresponding to an improvement from 247 ± 8 L CH4/kg VS to 317 ± 1 L CH4/kg VS. The enhanced removal of toxic metals further increased the methane production by another 18% to 360 ± 6 L CH4/kg VS (a total increase of 46%). The solids content of dewatered sludge increased from 14.6 ± 1.4% in the control to 18.2 ± 0.8%. A 4-log reduction for both total coliforms and E. coli was achieved. Overall, this study highlights the potential of acidification with low level nitrite addition as an effective and simple method achieving multiple improvements in terms of sludge management.

Nitrite addition to acidified sludge significantly improves digestibility, toxic metal removal, dewaterability and pathogen reduction

PubMed Central

Du, Fangzhou; Keller, Jürg; Yuan, Zhiguo; Batstone, Damien J.; Freguia, Stefano; Pikaar, Ilje

2016-01-01

Sludge management is a major issue for water utilities globally. Poor digestibility and dewaterability are the main factors determining the cost for sludge management, whereas pathogen and toxic metal concentrations limit beneficial reuse. In this study, the effects of low level nitrite addition to acidified sludge to simultaneously enhance digestibility, toxic metal removal, dewaterability and pathogen reduction were investigated. Waste activated sludge (WAS) from a full-scale waste water treatment plant was treated at pH 2 with 10 mg NO2−-N/L for 5 h. Biochemical methane potential tests showed an increase in the methane production of 28%, corresponding to an improvement from 247 ± 8 L CH4/kg VS to 317 ± 1 L CH4/kg VS. The enhanced removal of toxic metals further increased the methane production by another 18% to 360 ± 6 L CH4/kg VS (a total increase of 46%). The solids content of dewatered sludge increased from 14.6 ± 1.4% in the control to 18.2 ± 0.8%. A 4-log reduction for both total coliforms and E. coli was achieved. Overall, this study highlights the potential of acidification with low level nitrite addition as an effective and simple method achieving multiple improvements in terms of sludge management. PMID:28004811

Laboratory simulation reveals significant impacts of ocean acidification on microbial community composition and host-pathogen interactions between the blood clam and Vibrio harveyi.

PubMed

Zha, Shanjie; Liu, Saixi; Su, Wenhao; Shi, Wei; Xiao, Guoqiang; Yan, Maocang; Liu, Guangxu

2017-12-01

It has been suggested that climate change may promote the outbreaks of diseases in the sea through altering the host susceptibility, the pathogen virulence, and the host-pathogen interaction. However, the impacts of ocean acidification (OA) on the pathogen components of bacterial community and the host-pathogen interaction of marine bivalves are still poorly understood. Therefore, 16S rRNA high-throughput sequencing and host-pathogen interaction analysis between blood clam (Tegillarca granosa) and Vibrio harveyi were conducted in the present study to gain a better understanding of the ecological impacts of ocean acidification. The results obtained revealed a significant impact of ocean acidification on the composition of microbial community at laboratory scale. Notably, the abundance of Vibrio, a major group of pathogens to many marine organisms, was significantly increased under ocean acidification condition. In addition, the survival rate and haemolytic activity of V. harveyi were significantly higher in the presence of haemolymph of OA treated T. granosa, indicating a compromised immunity of the clam and enhanced virulence of V. harveyi under future ocean acidification scenarios. Conclusively, the results obtained in this study suggest that future ocean acidification may increase the risk of Vibrio pathogen infection for marine bivalve species, such as blood clams. Copyright © 2017 Elsevier Ltd. All rights reserved.

Estimating the probability of an extinction or major outbreak for an environmentally transmitted infectious disease.

PubMed

Lahodny, G E; Gautam, R; Ivanek, R

2015-01-01

Indirect transmission through the environment, pathogen shedding by infectious hosts, replication of free-living pathogens within the environment, and environmental decontamination are suspected to play important roles in the spread and control of environmentally transmitted infectious diseases. To account for these factors, the classic Susceptible-Infectious-Recovered-Susceptible epidemic model is modified to include a compartment representing the amount of free-living pathogen within the environment. The model accounts for host demography, direct and indirect transmission, replication of free-living pathogens in the environment, and removal of free-living pathogens by natural death or environmental decontamination. Based on the assumptions of the deterministic model, a continuous-time Markov chain model is developed. An estimate for the probability of disease extinction or a major outbreak is obtained by approximating the Markov chain with a multitype branching process. Numerical simulations illustrate important differences between the deterministic and stochastic counterparts, relevant for outbreak prevention, that depend on indirect transmission, pathogen shedding by infectious hosts, replication of free-living pathogens, and environmental decontamination. The probability of a major outbreak is computed for salmonellosis in a herd of dairy cattle as well as cholera in a human population. An explicit expression for the probability of disease extinction or a major outbreak in terms of the model parameters is obtained for systems with no direct transmission or replication of free-living pathogens.

Use of 1% peroxyacetic acid sanitizer in an air-mixing wash basin to remove bacterial pathogens from seeds.

PubMed

Rajkowski, Kathleen T; Ashurst, Kean

2009-11-01

To achieve the production of pathogen-free sprouts, there must be appropriate mixing of liquid sanitizer with the seeds to assure contact. Commercial treatments by irradiation or ozone gas of Salmonella spp. artificially inoculated seeds were compared, and these resulted in a 1 log reduction after all treatments. Use of peroxyacetic acid (1%) sanitizer on Salmonella spp. or Escherichia coli O157:H7 inoculated alfalfa seeds consistently resulted in a greater than 1 log reduction. In addition, during these studies debris was noted after the seeds were removed. Based on this observation, an air-mixing wash basin was developed for commercial use. Validation was done by commercial growers using 1% peroxyacetic acid sanitizer to wash seeds in the air-mixing basin, followed by sprouting the seeds. No positive or false-positive pathogen results were reported after the required testing of the sprout water (run-off during sprouting). Use of 1% peroxyacetic acid sanitizer in the air-mixing wash basin does provide the sprout grower an effective means of sanitizing sprout seeds.

Inactivation of Salmonella, Listeria monocytogenes and Enterococcus faecium NRRL B-2354 in a selection of low moisture foods.

PubMed

Rachon, Grzegorz; Peñaloza, Walter; Gibbs, Paul A

2016-08-16

The aims of this study were to obtain data on survival and heat resistance of cocktails of Salmonella, Listeria monocytogenes and the surrogate Enterococcus faecium (NRRL B-2354) in four low moisture foods (confectionery formulation, chicken meat powder, pet food and savoury seasoning) during storage before processing. Inoculated samples were stored at 16°C and cell viability examined at day 0, 3, 7 and 21. At each time point, the heat resistance at 80°C was determined. The purpose was to determine a suitable storage time of inoculated foods that can be applied in heat resistance studies or process validations with similar cell viability and heat resistance characteristics. The main inactivation study was carried out within 7days after inoculation, the heat resistance of each bacterial cocktail was evaluated in each low moisture food heated in thermal cells exposed to temperatures between 70 and 140°C. The Weibull model and the first order kinetics (D-value) were used to express inactivation data and calculate the heating time to achieve 5 log reduction at each temperature. Results showed that the pathogens Salmonella and L. monocytogenes and the surrogate E. faecium NRRL B-2354, can survive well (maximum reduction <0.8 log) in low moisture foods maintained at 16°C, as simulation of warehouse raw material storage in winter and before processing. The D80 value of the pathogens and surrogate did not significantly change during the 21day storage (p>0.05). The inactivation kinetics of the pathogens and surrogate at temperatures between 70 and 140°C, were different between each organism and product. E. faecium NRRL B-2354 was a suitable Salmonella surrogate for three of the low moisture foods studied, but not for the sugar-containing confectionery formulation. Heating low moisture food in moisture-tight environments (thermal cells) to 111.2, 105.3 or 111.8°C can inactivate 5 log of Salmonella, L. monocytogenes or E. faecium NRRL B-2354 respectively. Copyright © 2016 Elsevier B.V. All rights reserved.

Decreased waterborne pathogenic bacteria in an urban aquifer related to intense shallow geothermal exploitation.

PubMed

García-Gil, Alejandro; Gasco-Cavero, Samanta; Garrido, Eduardo; Mejías, Miguel; Epting, Jannis; Navarro-Elipe, Mercedes; Alejandre, Carmen; Sevilla-Alcaine, Elena

2018-08-15

The implications of intensive use of shallow geothermal energy resources in shallow urban aquifers are still not known for waterborne pathogens relevant to human health. Firstly, we hypothesized that waterborne enteric pathogens would be relatively increased in heated groundwater plumes. To prove this, microbiological sampling of 31 piezometers covering the domain of an urban groundwater body affected by microbiological contamination and energetically exploited by 70 groundwater heat pump systems was performed. Mean differences of pathogenic bacteria contents between impacted and non-impacted monitoring points were assessed with a two-tailed independent Student's t-test or Mann-Whitney U and correlation coefficients were also calculated. Surprisingly, the results obtained revealed a significant and generalized decrease in waterborne pathogen contents in thermally impacted piezometers compared to that of non-impacted piezometers. This decrease is hypothesized to be caused by a heat shock to bacteria within the heat exchangers. The statistically significant negative correlations obtained between waterborne pathogen counts and temperature could be explained by the spatial distribution of the bacteria, finding that bacteria start to recover with increasing distance from the injection point. Also, different behavior groups fitting exponential regression models were found for the bacteria species studied, justified by the different presence and influence of several aquifer parameters and major, minor and trace elements studied, as well as the coexistence with other bacteria species. The results obtained from this work reinforce the concept of shallow geothermal resources as a clean energy source, as they could also provide the basis to control the pathogenic bacteria contents in groundwater bodies. Copyright © 2018 Elsevier B.V. All rights reserved.

Characterization of SiaA, a streptococcal heme-binding protein associated with a heme ABC transport system.

PubMed

Sook, Brian R; Block, Darci R; Sumithran, Suganya; Montañez, Griselle E; Rodgers, Kenton R; Dawson, John H; Eichenbaum, Zehava; Dixon, Dabney W

2008-02-26

Many pathogenic bacteria require heme and obtain it from their environment. Heme transverses the cytoplasmic membrane via an ATP binding cassette (ABC) pathway. Although a number of heme ABC transport systems have been described in pathogenic bacteria, there is as yet little biophysical characterization of the proteins in these systems. The sia (hts) gene cluster encodes a heme ABC transporter in the Gram positive Streptococcus pyogenes. The lipoprotein-anchored heme binding protein (HBP) of this transporter is SiaA (HtsA). In the current study, resonance Raman (rR), magnetic circular dichroism (MCD), and nuclear magnetic resonance (NMR) spectroscopies were used to determine the coordination state and spin state of both the ferric and ferrous forms of this protein. Identifiers from these techniques suggest that the heme is six-coordinate and low-spin in both oxidation states of the protein, with methionine and histidine as axial ligands. SiaA has a pKa of 9.7 +/- 0.1, attributed to deprotonation of the axial histidine. Guanidinium titration studies show that the ferric state is less stable than the ferrous state, with DeltaG(H2O) values for the oxidized and reduced proteins of 7.3 +/- 0.8 and 16.0 +/- 3.6 kcal mol-1, respectively. The reductive and oxidative midpoint potentials determined via spectroelectrochemistry are 83 +/- 3 and 64 +/- 3 mV, respectively; the irreversibility of heme reduction suggests that redox cycling of the heme is coupled to a kinetically sluggish change in structure or conformation. The biophysical characterization described herein will significantly advance our understanding of structure-function relationships in HBP.

Risk Reduction Modeling of High Pathogenicity Avian Influenza Virus Titers in Nonpasteurized Liquid Egg Obtained from Infected but Undetected Chicken Flocks.

PubMed

Weaver, J Todd; Malladi, Sasidhar; Spackman, Erica; Swayne, David E

2015-11-01

Control of highly pathogenic avian influenza (HPAI) outbreaks in poultry has traditionally involved the establishment of disease containment zones, where poultry products are only permitted to move from within a zone under permit. Nonpasteurized liquid egg (NPLE) is one such commodity for which movements may be permitted, considering inactivation of HPAI virus via pasteurization. Active surveillance testing at the flock level, using targeted matrix gene real-time reversed transcriptase-polymerase chain reaction testing (RRT-PCR) has been incorporated into HPAI emergency response plans as the primary on-farm diagnostic test procedure to detect HPAI in poultry and is considered to be a key risk mitigation measure. To inform decisions regarding the potential movement of NPLE to a pasteurization facility, average HPAI virus concentrations in NPLE produced from a HPAI virus infected, but undetected, commercial table-egg-layer flock were estimated for three HPAI virus strains using quantitative simulation models. Pasteurization under newly proposed international design standards (5 log10 reduction) is predicted to inactivate HPAI virus in NPLE to a very low concentration of less than 1 embryo infectious dose (EID)50 /mL, considering the predicted virus titers in NPLE from a table-egg flock under active surveillance. Dilution of HPAI virus from contaminated eggs in eggs from the same flock, and in a 40,000 lb tanker-truck load of NPLE containing eggs from disease-free flocks was also considered. Risk assessment can be useful in the evaluation of commodity-specific risk mitigation measures to facilitate safe trade in animal products from countries experiencing outbreaks of highly transmissible animal diseases. © 2015 Society for Risk Analysis.

New connection method for isolating and disinfecting intraluminal path during peritoneal dialysis solution-exchange procedures.

PubMed

Grabowy, R S; Kelley, R; Richter, S G; Bousquet, G G; Carr, K L

1998-01-01

Microbiological data have been collected on the performance of a new method of isolating and disinfecting the intraluminal path at the connect/disconnect site of a peritoneal dialysis (PD)-exchange pathway. High-temperature moist-heat (HTMH) disinfection is accomplished by a new device that uses microwave energy to heat the solution contained in the pressure-tight inner lumen of PD connector pairs between the transfer-set connector-clamp and the bag-connector break-away seal. An 85 degrees C (S.D. = 2.4 degrees C, n = 10) rise in solution temperature is seen in 12 seconds, thus yielding temperatures under pressure well over 100 degrees C with starting temperatures of 25 degrees C. Connector pairs were prepared by inoculation of a solution suspension containing at least 10(6) colony-forming units (CFU) of a test micro-organism. Approximately 0.4 mL of solution was contained within the mated connector pair. Using standard D-value determination methods, data were obtained for surviving organisms versus five exposure times and a positive control to obtain a population reduction curve. Four micro-organisms (S. epidermidis, P. aeruginosa, C. albicans, and A. niger) recognized to be among the most prevalent or problematic in causing peritonitis were tested. After microwave heating, the treated solution was aseptically withdrawn from the connector pair using a needle and syringe, plated in growth media, and incubated. Population counts of CFUs after incubation were used to establish survival curves. Results showed a tenfold population reduction in less than 3 seconds for all organisms tested. A 30-second cycle time safely achieves a > 10(8) population-reduction for bacteria and yeast organisms, and a > 10(7) population reduction for fungi. One potential benefit of using this new intraluminal disinfection method is that it may help reduce peritonitis resulting from the even more problematic pathogens such as the gram-negative bacteria and fungal organisms.

Control of tick infestations and pathogen prevalence in cattle and sheep farms vaccinated with the recombinant Subolesin-Major Surface Protein 1a chimeric antigen

PubMed Central

2014-01-01

Background Despite the use of chemical acaricides, tick infestations continue to affect animal health and production worldwide. Tick vaccines have been proposed as a cost-effective and environmentally friendly alternative for tick control. Vaccination with the candidate tick protective antigen, Subolesin (SUB), has been shown experimentally to be effective in controlling vector infestations and pathogen infection. Furthermore, Escherichia coli membranes containing the chimeric antigen composed of SUB fused to Anaplasma marginale Major Surface Protein 1a (MSP1a) (SUB-MSP1a) were produced using a simple low-cost process and proved to be effective for the control of cattle tick, Rhipicephalus (Boophilus) microplus and R. annulatus infestations in pen trials. In this research, field trials were conducted to characterize the effect of vaccination with SUB-MSP1a on tick infestations and the prevalence of tick-borne pathogens in a randomized controlled prospective study. Methods Two cattle and two sheep farms with similar geographical locations and production characteristics were randomly assigned to control and vaccinated groups. Ticks were collected, counted, weighed and classified and the prevalence of tick-borne pathogens at the DNA and serological levels were followed for one year prior to and 9 months after vaccination. Results Both cattle and sheep developed antibodies against SUB in response to vaccination. The main effect of the vaccine in cattle was the 8-fold reduction in the percent of infested animals while vaccination in sheep reduced tick infestations by 63%. Female tick weight was 32-55% lower in ticks collected from both vaccinated cattle and sheep when compared to controls. The seroprevalence of Babesia bigemina was lower by 30% in vaccinated cattle, suggesting a possible role for the vaccine in decreasing the prevalence of this tick-borne pathogen. The effect of the vaccine in reducing the frequency of one A. marginale msp4 genotype probably reflected the reduction in the prevalence of a tick-transmitted strain as a result of the reduction in the percent of tick-infested cattle. Conclusions These data provide evidence of the dual effect of a SUB-based vaccine for controlling tick infestations and pathogen infection/transmission and provide additional support for the use of the SUB-MSP1a vaccine for tick control in cattle and sheep. PMID:24398155

Control of tick infestations and pathogen prevalence in cattle and sheep farms vaccinated with the recombinant Subolesin-Major Surface Protein 1a chimeric antigen.

PubMed

Torina, Alessandra; Moreno-Cid, Juan A; Blanda, Valeria; Fernández de Mera, Isabel G; de la Lastra, José M Pérez; Scimeca, Salvatore; Blanda, Marcellocalogero; Scariano, Maria Elena; Briganò, Salvatore; Disclafani, Rosaria; Piazza, Antonio; Vicente, Joaquín; Gortázar, Christian; Caracappa, Santo; Lelli, Rossella Colomba; de la Fuente, José

2014-01-08

Despite the use of chemical acaricides, tick infestations continue to affect animal health and production worldwide. Tick vaccines have been proposed as a cost-effective and environmentally friendly alternative for tick control. Vaccination with the candidate tick protective antigen, Subolesin (SUB), has been shown experimentally to be effective in controlling vector infestations and pathogen infection. Furthermore, Escherichia coli membranes containing the chimeric antigen composed of SUB fused to Anaplasma marginale Major Surface Protein 1a (MSP1a) (SUB-MSP1a) were produced using a simple low-cost process and proved to be effective for the control of cattle tick, Rhipicephalus (Boophilus) microplus and R. annulatus infestations in pen trials. In this research, field trials were conducted to characterize the effect of vaccination with SUB-MSP1a on tick infestations and the prevalence of tick-borne pathogens in a randomized controlled prospective study. Two cattle and two sheep farms with similar geographical locations and production characteristics were randomly assigned to control and vaccinated groups. Ticks were collected, counted, weighed and classified and the prevalence of tick-borne pathogens at the DNA and serological levels were followed for one year prior to and 9 months after vaccination. Both cattle and sheep developed antibodies against SUB in response to vaccination. The main effect of the vaccine in cattle was the 8-fold reduction in the percent of infested animals while vaccination in sheep reduced tick infestations by 63%. Female tick weight was 32-55% lower in ticks collected from both vaccinated cattle and sheep when compared to controls. The seroprevalence of Babesia bigemina was lower by 30% in vaccinated cattle, suggesting a possible role for the vaccine in decreasing the prevalence of this tick-borne pathogen. The effect of the vaccine in reducing the frequency of one A. marginale msp4 genotype probably reflected the reduction in the prevalence of a tick-transmitted strain as a result of the reduction in the percent of tick-infested cattle. These data provide evidence of the dual effect of a SUB-based vaccine for controlling tick infestations and pathogen infection/transmission and provide additional support for the use of the SUB-MSP1a vaccine for tick control in cattle and sheep.

CHEMICAL CHARACTERIZATION AND EVALUATION OF ANTIBACTERIAL, ANTIFUNGAL, ANTIMYCOBACTERIAL, AND CYTOTOXIC ACTIVITIES OF Talinum paniculatum

PubMed Central

REIS, Luis F.C. DOS; CERDEIRA, Cláudio D.; PAULA, Bruno F. DE; da SILVA, Jeferson J.; COELHO, Luiz F.L.; SILVA, Marcelo A.; MARQUES, Vanessa B.B.; CHAVASCO, Jorge K.; ALVES-DA-SILVA, Geraldo

2015-01-01

SUMMARY In this study, the bioactivity of Talinum paniculatum was evaluated, a plant widely used in folk medicine. The extract from the T. paniculatum leaves (LE) was obtained by percolation with ethanol-water and then subjecting it to liquid-liquid partitions, yielding hexane (HX), ethyl acetate (EtOAc), butanol (BuOH), and aqueous (Aq) fractions. Screening for antimicrobial activity of the LE and its fractions was evaluated in vitro through broth microdilution method, against thirteen pathogenic and non-pathogenic microorganisms, and the antimycobacterial activity was performed through agar diffusion assay. The cytotoxic concentrations (CC90) for LE, HX, and EtOAc were obtained on BHK-21 cells by using MTT reduction assay. The LE showed activity against Serratia marcescens and Staphylococcus aureus, with Minimum Inhibitory Concentration (MIC) values of 250 and 500 µg/mL, respectively. Furthermore, HX demonstrated outstanding activity against Micrococcus luteusand Candida albicans with a MIC of 31.2 µg/mL in both cases. The MIC for EtOAc also was 31.2 µg/mL against Escherichia coli. Conversely, BuOH and Aq were inactive against all tested microorganisms and LE proved inactive against Mycobacterium tuberculosis and Mycobacterium bovis as well. Campesterol, stigmasterol, and sitosterol were the proposed structures as main compounds present in the EF and HX/EtOAc fractions, evidenced by mass spectrometry. Therefore, LE, HX, and EtOAc from T. paniculatum showed potential as possible sources of antimicrobial compounds, mainly HX, for presenting low toxicity on BHK-21 cells with excellent Selectivity Index (SI = CC90/MIC) of 17.72 against C. albicans. PMID:26603226

Thionin-like peptides from Capsicum annuum fruits with high activity against human pathogenic bacteria and yeasts.

PubMed

Taveira, Gabriel B; Mathias, Luciana S; da Motta, Olney V; Machado, Olga L T; Rodrigues, Rosana; Carvalho, André O; Teixeira-Ferreira, André; Perales, Jonas; Vasconcelos, Ilka M; Gomes, Valdirene M

2014-01-01

Plants defend themselves against pathogens with production of antimicrobial peptides (AMPs). Herein we describe the discovery of a new antifungal and antibacterial peptide from fruits of Capsicum annuum that showed similarity to an already well characterized family of plant AMPs, thionins. Other fraction composed of two peptides, in which the major peptide also showed similarity to thionins. Among the obtained fractions, fraction 1, which is composed of a single peptide of 7 kDa, was sequenced by Edman method and its comparative sequence analysis in database (nr) showed similarity to thionin-like peptides. Tests against microorganisms, fraction 1 presented inhibitory activity to the cells of yeast Saccharomyces cerevisiae, Candida albicans, and Candida tropicalis and caused growth reduction to the bacteria species Escherichia coli and Pseudomonas aeruginosa. Fraction 3 caused inhibitory activity only for C. albicans and C. tropicalis. This fraction was composed of two peptides of ∼7 and 10 kDa, and the main protein band correspondent to the 7 kDa peptide, also showed similarity to thionins. This plasma membrane permeabilization assay demonstrates that the peptides present in the fractions 1 and 3 induced changes in the membranes of all yeast strains, leading to their permeabilization. Fraction 1 was capable of inhibiting acidification of the medium of glucose-induced S. cerevisiae cells 78% after an incubation time of 30 min, and opposite result was obtained for C. albicans. Experiments demonstrate that the fraction 1 and 3 were toxic and induced changes in the membranes of all yeast strains, leading to their permeabilization. Copyright © 2013 Wiley Periodicals, Inc.

Host-pathogen dynamics under sterilizing pathogens and fecundity-longevity trade-off in hosts.

PubMed

Janoušková, Eva; Berec, Luděk

2018-08-07

Infectious diseases are known to regulate population dynamics, an observation that underlies the use of pathogens as control agents of unwanted populations. Sterilizing rather than lethal pathogens are often suggested so as to avoid unnecessary suffering of the infected hosts. Until recently, models used to assess plausibility of pathogens as potential pest control agents have not included a possibility that reduced fecundity of the infected individuals may save their energy expenditure on reproduction and thus increase their longevity relative to the susceptible ones. Here, we develop a model of host-pathogen interaction that builds on this idea. We analyze the model for a variety of infection transmission functions, revealing that the indirect effect of sterilizing pathogens on mortality of the infected hosts, mediated by a fecundity-longevity trade-off, may cause hosts at endemic equilibria to attain densities higher than when there is no effect of pathogens on host mortality. On the other hand, an opposite outcome occurs when the fecundity-longevity trade-off is concave or when the degree of fecundity reduction by the pathogen is high enough. This points to a possibility that using sterilizing pathogens as agents of pest control may actually be less effective than previously thought, the more so since we also suggest that if sexual selection acts on the host species then the presence of sterilizing pathogens may even enhance host densities above the levels achieved without infection. Copyright © 2018 Elsevier Ltd. All rights reserved.

[EXPERIENCE OF STUDY AND POSSIBLE WAYS OF ELIMINATION OF FALSE POSITIVE AND FALSE NEGATIVE RESULTS DURING EXECUTION OF POLYMERASE CHAIN REACTION ON AN EXAMPLE OF JUNIN VIRUS RNA DETECTION].

PubMed

Sizikova, T E; Lebedev, V N; Pantyukhov, V B; Borisevich, S V; Merkulov, V A

2015-01-01

Experience of study and possible ways of elimination of false positive and false negative results during execution of polymerase chain reaction on an example of Junin virus RNA detection. MATERIALSS AND METHODS: Junin virus--causative agent of Argentine hemorrhagic fever (AHF) strain XJpR37/5787 was obtained from the State collection of pathogenicity group I causative agents of the 48th Central Research Institute. Reagent kit for detection of Junin virus RNA by RT-PCR was developed in the Institute and consists of 4 sets: for isolation of RNA, execution of reverse-transcription reaction, execution of PCR and electrophoretic detection of PCR products. RT-PCR was carried out by a standard technique. Continuous cell cultures of African green monkey Vero B, GMK-AH-1(D) were obtained from the museum of cell culture department of the Centre. An experimental study of the effect of various factors of impact on the sample under investigation ("thawing-freezing", presence of formaldehyde, heparin) on the obtaining of false negative results during Junin virus RNA detection by using RT-PCR was studied. Addition of 0.01% heparin to the samples was shown to completely inhibit PCR. Addition of 0.05% formaldehyde significantly reduces sensitivity of the method. A possibility of reduction of analysis timeframe from 15 to 5 days was shown during detection of the causative agent in samples with low concentration of the latter by growing the samples and subsequent analysis of the material obtained by using RT-PCR. During detection of causative agent by using RT-PCR false negative results could appear in the presence of formaldehyde and heparin in the sample. A possibility of elimination of false negative PCR results due to concentration of the causative agent in the sample under investigation at a level below sensitivity threshold was shown on the example of Junin virus RNA detection by using growing of the pathogen in appropriate accumulation system with subsequent analysis of the material obtained using PCR.

Celecoxib Enhances the Efficacy of Low-Dose Antibiotic Treatment against Polymicrobial Sepsis in Mice and Clinical Isolates of ESKAPE Pathogens

PubMed Central

Annamanedi, Madhavi; Varma, Gajapati Y. N.; Anuradha, K.; Kalle, Arunasree M.

2017-01-01

Treatment of multidrug resistant bacterial infections has been a great challenge globally. Previous studies including our study have highlighted the use of celecoxib, a non-steroidal anti-inflammatory drug in combination with antibiotic has decreased the minimal inhibitory concentration to limit Staphylococcus aureus infection. However, the efficacy of this combinatorial treatment against various pathogenic bacteria is not determined. Therefore, we have evaluated the potential use of celecoxib in combination with low doses of antibiotic in limiting Gram-positive and Gram-negative bacteria in vivo in murine polymicrobial sepsis developed by cecum ligation and puncture (CLP) method and against clinically isolated human ESKAPE pathogens (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species). The in vivo results clearly demonstrated a significant reduction in the bacterial load in different organs and in the inflammatory markers such as COX-2 and NF-κB via activation of SIRT1 in mice treated with imipenem, a choice of antibiotic for polymicrobial sepsis treatment. Combinatorial treatment of ampicillin and celecoxib was effective on clinical isolates of ESKAPE pathogens, 45% of tested clinical isolates showed more than 50% reduction in the colony forming units when compared to ampicillin alone. In conclusion, this non-traditional treatment strategy might be effective in clinic to reduce the dose of antibiotic to treat drug-resistant bacterial infections. PMID:28533769

Celecoxib Enhances the Efficacy of Low-Dose Antibiotic Treatment against Polymicrobial Sepsis in Mice and Clinical Isolates of ESKAPE Pathogens.

PubMed

Annamanedi, Madhavi; Varma, Gajapati Y N; Anuradha, K; Kalle, Arunasree M

2017-01-01

Treatment of multidrug resistant bacterial infections has been a great challenge globally. Previous studies including our study have highlighted the use of celecoxib, a non-steroidal anti-inflammatory drug in combination with antibiotic has decreased the minimal inhibitory concentration to limit Staphylococcus aureus infection. However, the efficacy of this combinatorial treatment against various pathogenic bacteria is not determined. Therefore, we have evaluated the potential use of celecoxib in combination with low doses of antibiotic in limiting Gram-positive and Gram-negative bacteria in vivo in murine polymicrobial sepsis developed by cecum ligation and puncture (CLP) method and against clinically isolated human ESKAPE pathogens ( Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa , and Enterobacter species). The in vivo results clearly demonstrated a significant reduction in the bacterial load in different organs and in the inflammatory markers such as COX-2 and NF-κB via activation of SIRT1 in mice treated with imipenem, a choice of antibiotic for polymicrobial sepsis treatment. Combinatorial treatment of ampicillin and celecoxib was effective on clinical isolates of ESKAPE pathogens, 45% of tested clinical isolates showed more than 50% reduction in the colony forming units when compared to ampicillin alone. In conclusion, this non-traditional treatment strategy might be effective in clinic to reduce the dose of antibiotic to treat drug-resistant bacterial infections.

Effect of atmospheric pressure plasma on inactivation of pathogens inoculated onto bacon using two different gas compositions.

PubMed

Kim, Binna; Yun, Hyejeong; Jung, Samooel; Jung, Yeonkook; Jung, Heesoo; Choe, Wonho; Jo, Cheorun

2011-02-01

Atmospheric pressure plasma (APP) is an emerging non-thermal pasteurization method for the enhancement of food safety. In this study, the effect of APP on the inactivation of pathogens inoculated onto bacon was observed. Sliced bacon was inoculated with Listeria monocytogenes (KCTC 3596), Escherichia coli (KCTC 1682), and Salmonella Typhimurium (KCTC 1925). The samples were treated with APP at 75, 100, and 125 W of input power for 60 and 90 s. Two gases, helium (10 lpm) or a mixture of helium and oxygen, (10 lpm and 10 sccm, respectively) were used for the plasma generation. Plasma with helium could only reduce the number of inoculated pathogens by about 1-2 Log cycles. On the other hand, the helium/oxygen gas mixture was able to achieve microbial reduction of about 2-3 Log cycles. The number of total aerobic bacteria showed 1.89 and 4.58 decimal reductions after plasma treatment with helium and the helium/oxygen mixture, respectively. Microscopic observation of the bacon after plasma treatment did not find any significant changes, except that the L∗-value of the bacon surface was increased. These results clearly indicate that APP treatment is effective for the inactivation of the three pathogens used in this study, although further investigation is needed for elucidating quality changes after treatment. Copyright © 2010 Elsevier Ltd. All rights reserved.

Sepsis and Hemocyte Loss in Honey Bees (Apis mellifera) Infected with Serratia marcescens Strain Sicaria.

PubMed

Burritt, Nancy L; Foss, Nicole J; Neeno-Eckwall, Eric C; Church, James O; Hilger, Anna M; Hildebrand, Jacob A; Warshauer, David M; Perna, Nicole T; Burritt, James B

2016-01-01

Global loss of honey bee colonies is threatening the human food supply. Diverse pathogens reduce honey bee hardiness needed to sustain colonies, especially in winter. We isolated a free-living Gram negative bacillus from hemolymph of worker honey bees (Apis mellifera) found separated from winter clusters. In some hives, greater than 90% of the dying bees detached from the winter cluster were found to contain this bacterium in their hemolymph. Throughout the year, the same organism was rarely found in bees engaged in normal hive activities, but was detected in about half of Varroa destructor mites obtained from colonies that housed the septic bees. Flow cytometry of hemolymph from septic bees showed a significant reduction of plasmatocytes and other types of hemocytes. Interpretation of the16S rRNA sequence of the bacterium indicated that it belongs to the Serratia genus of Gram-negative Gammaproteobacteria, which has not previously been implicated as a pathogen of adult honey bees. Complete genome sequence analysis of the bacterium supported its classification as a novel strain of Serratia marcescens, which was designated as S. marcescens strain sicaria (Ss1). When compared with other strains of S. marcescens, Ss1 demonstrated several phenotypic and genetic differences, including 65 genes not previously found in other Serratia genomes. Some of the unique genes we identified in Ss1 were related to those from bacterial insect pathogens and commensals. Recovery of this organism extends a complex pathosphere of agents which may contribute to failure of honey bee colonies.

Identification, characterization and phylogenetic analysis of antifungal Trichoderma from tomato rhizosphere.

PubMed

Rai, Shalini; Kashyap, Prem Lal; Kumar, Sudheer; Srivastava, Alok Kumar; Ramteke, Pramod W

2016-01-01

The use of Trichoderma isolates with efficient antagonistic activity represents a potentially effective and alternative disease management strategy to replace health hazardous chemical control. In this context, twenty isolates were obtained from tomato rhizosphere and evaluated by their antagonistic activity against four fungal pathogens ( Fusarium oxysporum f. sp. lycopersici , Alternaria alternata , Colletotrichum gloeosporoides and Rhizoctonia solani ). The production of extracellular cell wall degrading enzymes of tested isolates was also measured. All the isolates significantly reduced the mycelial growth of tested pathogens but the amount of growth reduction varied significantly as well. There was a positive correlation between the antagonistic capacity of Trichoderma isolates towards fungal pathogens and their lytic enzyme production. The Trichoderma isolates were initially sorted according to morphology and based on the translation elongation factor 1-α gene sequence similarity, the isolates were designated as Trichoderma harzianum , T. koningii , T. asperellum , T. virens and T. viride . PCA analysis explained 31.53, 61.95, 62.22 and 60.25% genetic variation among Trichoderma isolates based on RAPD, REP-, ERIC- and BOX element analysis, respectively. ERG - 1 gene, encoding a squalene epoxidase has been used for the first time for diversity analysis of antagonistic Trichoderma from tomato rhizosphere. Phylogenetic analysis of ERG -1 gene sequences revealed close relatedness of ERG -1sequences with earlier reported sequences of Hypocrea lixii , T. arundinaceum and T. reesei. However, ERG -1 gene also showed heterogeneity among some antagonistic isolates and indicated the possibility of occurrence of squalene epoxidase driven triterpene biosynthesis as an alternative biocontrol mechanism in Trichoderma species.

Sepsis and Hemocyte Loss in Honey Bees (Apis mellifera) Infected with Serratia marcescens Strain Sicaria

PubMed Central

Burritt, Nancy L.; Foss, Nicole J.; Neeno-Eckwall, Eric C.; Church, James O.; Hildebrand, Jacob A.; Warshauer, David M.; Perna, Nicole T.; Burritt, James B.

2016-01-01

Global loss of honey bee colonies is threatening the human food supply. Diverse pathogens reduce honey bee hardiness needed to sustain colonies, especially in winter. We isolated a free-living Gram negative bacillus from hemolymph of worker honey bees (Apis mellifera) found separated from winter clusters. In some hives, greater than 90% of the dying bees detached from the winter cluster were found to contain this bacterium in their hemolymph. Throughout the year, the same organism was rarely found in bees engaged in normal hive activities, but was detected in about half of Varroa destructor mites obtained from colonies that housed the septic bees. Flow cytometry of hemolymph from septic bees showed a significant reduction of plasmatocytes and other types of hemocytes. Interpretation of the16S rRNA sequence of the bacterium indicated that it belongs to the Serratia genus of Gram-negative Gammaproteobacteria, which has not previously been implicated as a pathogen of adult honey bees. Complete genome sequence analysis of the bacterium supported its classification as a novel strain of Serratia marcescens, which was designated as S. marcescens strain sicaria (Ss1). When compared with other strains of S. marcescens, Ss1 demonstrated several phenotypic and genetic differences, including 65 genes not previously found in other Serratia genomes. Some of the unique genes we identified in Ss1 were related to those from bacterial insect pathogens and commensals. Recovery of this organism extends a complex pathosphere of agents which may contribute to failure of honey bee colonies. PMID:28002470

Components of a Rice-Oilseed Rape Production System Augmented with Trichoderma sp. Tri-1 Control Sclerotinia sclerotiorum on Oilseed Rape.

PubMed

Hu, Xiaojia; Roberts, Daniel P; Xie, Lihua; Maul, Jude E; Yu, Changbing; Li, Yinshui; Zhang, Yinbo; Qin, Lu; Liao, Xing

2015-10-01

Sclerotinia sclerotiorum causes serious yield losses on many crops throughout the world. A multicomponent treatment that consisted of the residual rice straw remaining after rice harvest and Trichoderma sp. Tri-1 (Tri-1) formulated with the oilseed rape seedcake fertilizer was used in field soil infested with S. sclerotiorum. This treatment resulted in oilseed rape seed yield that was significantly greater than the nontreated control or when the fungicide carbendizem was used in the presence of this pathogen in field trials. Yield data suggested that the rice straw, oilseed rape seedcake, and Tri-1 components of this treatment all contributed incrementally. Similar treatment results were obtained regarding reduction in disease incidence. Slight improvements in yield and disease incidence were detected when this multicomponent treatment was combined with a fungicide spray. Inhibition of sclerotial germination by this multicomponent treatment trended greater than the nontreated control at 90, 120, and 150 days in field studies but was not significantly different from this control. This multicomponent treatment resulted in increased yield relative to the nontreated control in the absence of pathogen in a greenhouse pot study, while the straw alone and the straw plus oilseed rape seedcake treatments did not; suggesting that Tri-1 was capable of promoting growth. Experiments reported here indicate that a treatment containing components of a rice-oilseed rape production system augmented with Tri-1 can control S. sclerotiorum on oilseed rape, be used in integrated strategies containing fungicide sprays for control of this pathogen, and promote plant growth.

Antagonistic and Quantitative Assessment of Indigenous Lactic acid Bacteria in Different Varieties of Ogi against Gastrointestinal Pathogens

PubMed Central

Afolayan, Ayorinde Oluwatobiloba; Ayeni, Funmilola Abidemi; Ruppitsch, Werner

2017-01-01

Introduction Ogi is a popular fermented cereal gruel consumed mainly in the western part of Nigeria. Traditionally, uncooked Ogi is normally administered to diarrhoea patients to reduce the frequency of stooling. This study was therefore undertaken to identify, quantify and determine the antimicrobial properties of lactic acid bacteria (LAB) isolated from Ogi. Methods The Ogi samples (Yellow, white, sorghum) were obtained from different market in Ibadan, Nigeria and Ogi control (cooked, uncooked and Omidun) were prepared with the viable counts of bacteria monitored over 5 days period. LAB were isolated from the varieties and identified by partial sequencing of 16S rRNA gene. The antimicrobial activities of the cell free supernatant (CFS) and the viable cells of the isolated LAB against Escherichia coli EC004, Salmonella sp. SS11, Shigella sp. SS10 were investigated by agar diffusion assay, agar overlay method, and coculture growth studies. Results Omidun had the highest LAB count while cooked ogi has the lowest LAB count. Weissella paramesenteroides , L. brevis, L. rossiae, L. fermentum, L. plantarum, Acetobacter pasteurianus, Paenibacillus sp. and Bacillus sp. were isolated from Ogi in this study. Large zone of inhibition (11≤x≤20) was observed with CFS against Salmonella sp. SS11 and Shigella sp. SS10 and also the overlay method. Coculture studies of Weissella paramesenteroides, Lactobacillus fermentum, and L. plantarum with Salmonella sp. SS11 showed a 5-8 log reduction of the pathogens' growth after 24 hours as compared with the control. Conclusion Ogi and its contents have antimicrobial properties against pathogenic organisms. PMID:28748023

Shared and distinct mechanisms of iron acquisition by bacterial and fungal pathogens of humans

PubMed Central

Caza, Mélissa; Kronstad, James W.

2013-01-01

Iron is the most abundant transition metal in the human body and its bioavailability is stringently controlled. In particular, iron is tightly bound to host proteins such as transferrin to maintain homeostasis, to limit potential damage caused by iron toxicity under physiological conditions and to restrict access by pathogens. Therefore, iron acquisition during infection of a human host is a challenge that must be surmounted by every successful pathogenic microorganism. Iron is essential for bacterial and fungal physiological processes such as DNA replication, transcription, metabolism, and energy generation via respiration. Hence, pathogenic bacteria and fungi have developed sophisticated strategies to gain access to iron from host sources. Indeed, siderophore production and transport, iron acquisition from heme and host iron-containing proteins such as hemoglobin and transferrin, and reduction of ferric to ferrous iron with subsequent transport are all strategies found in bacterial and fungal pathogens of humans. This review focuses on a comparison of these strategies between bacterial and fungal pathogens in the context of virulence and the iron limitation that occurs in the human body as a mechanism of innate nutritional defense. PMID:24312900

Survival of Listeria monocytogenes on a conveyor belt material with or without antimicrobial additives.

PubMed

Chaitiemwong, N; Hazeleger, W C; Beumer, R R

2010-08-15

Survival of Listeria monocytogenes on a conveyor belt material with or without antimicrobial additives, in the absence or presence of food debris from meat, fish and vegetables and at temperatures of 10, 25 and 37 degrees C was investigated. The pathogen survived best at 10 degrees C, and better at 25 degrees C than at 37 degrees C on both conveyor belt materials. The reduction in the numbers of the pathogen on belt material with antimicrobial additives in the first 6h at 10 degrees C was 0.6 log unit, which was significantly higher (P<0.05) than the reduction of 0.2 log unit on belt material without additives. Reductions were significantly less (P<0.05) in the presence of food residue. At 37 degrees C and 20% relative humidity, large decreases in the numbers of the pathogen on both conveyor belt materials during the first 6h were observed. Under these conditions, there was no obvious effect of the antimicrobial substances. However, at 25 degrees C and 10 degrees C and high humidity (60-75% rh), a rapid decrease in bacterial numbers on the belt material with antimicrobial substances was observed. Apparently the reduction in numbers of L. monocytogenes on belt material with antimicrobial additives was greater than on belt material without additives only when the surfaces were wet. Moreover, the presence of food debris neutralized the effect of the antimicrobials. The results suggest that the antimicrobial additives in conveyor belt material could help to reduce numbers of microorganisms on belts at low temperatures when food residues are absent and belts are not rapidly dried. Copyright 2010 Elsevier B.V. All rights reserved.

Anthracnose of lucky bamboo Dracaena sanderiana caused by the fungus Colletotrichum dracaenophilum in Egypt.

PubMed

Morsy, Ahmed A; Elshahawy, Ibrahim E

2016-05-01

Dracaena sanderiana, of the family Liliaceae, is among the ornamental plants most frequently imported into Egypt. Typical anthracnose symptoms were observed on the stems of imported D. sanderiana samples. The pathogen was isolated, demonstrated to be pathogenic based on Koch's rule and identified as Colletotrichum dracaenophilum. The optimum temperature for its growth ranges from 25 to 30 °C, maintained for 8 days. Kemazed 50% wettable powder (WP) was the most effective fungicide against the pathogen, as no fungal growth was observed over 100 ppm. The biocontrol agents Trichoderma harzianum and Trichoderma viride followed by Bacillus subtilis and Bacillus pumilus caused the highest reduction in fungal growth. To the best of our knowledge, this report describes the first time that this pathogen was observed on D. sanderiana in Egypt.

Anthracnose of lucky bamboo Dracaena sanderiana caused by the fungus Colletotrichum dracaenophilum in Egypt

PubMed Central

Morsy, Ahmed A.; Elshahawy, Ibrahim E.

2016-01-01

Dracaena sanderiana, of the family Liliaceae, is among the ornamental plants most frequently imported into Egypt. Typical anthracnose symptoms were observed on the stems of imported D. sanderiana samples. The pathogen was isolated, demonstrated to be pathogenic based on Koch’s rule and identified as Colletotrichum dracaenophilum. The optimum temperature for its growth ranges from 25 to 30 °C, maintained for 8 days. Kemazed 50% wettable powder (WP) was the most effective fungicide against the pathogen, as no fungal growth was observed over 100 ppm. The biocontrol agents Trichoderma harzianum and Trichoderma viride followed by Bacillus subtilis and Bacillus pumilus caused the highest reduction in fungal growth. To the best of our knowledge, this report describes the first time that this pathogen was observed on D. sanderiana in Egypt. PMID:27222738

Removal of human pathogenic viruses in a down-flow hanging sponge (DHS) reactor treating municipal wastewater and health risks associated with utilization of the effluent for agricultural irrigation.

PubMed

Kobayashi, Naohiro; Oshiki, Mamoru; Ito, Toshihiro; Segawa, Takahiro; Hatamoto, Masashi; Kato, Tsuyoshi; Yamaguchi, Takashi; Kubota, Kengo; Takahashi, Masanobu; Iguchi, Akinori; Tagawa, Tadashi; Okubo, Tsutomu; Uemura, Shigeki; Harada, Hideki; Motoyama, Toshiki; Araki, Nobuo; Sano, Daisuke

2017-03-01

A down-flow hanging sponge (DHS) reactor has been developed as a cost-effective wastewater treatment system that is adaptable to local conditions in low-income countries. A pilot-scale DHS reactor previously demonstrated stable reduction efficiencies for chemical oxygen demand (COD) and ammonium nitrogen over a year at ambient temperature, but the pathogen reduction efficiency of the DHS reactor has yet to be investigated. In the present study, the reduction efficiency of a pilot-scale DHS reactor fed with municipal wastewater was investigated for 10 types of human pathogenic viruses (norovirus GI, GII and GIV, aichivirus, astrovirus, enterovirus, hepatitis A and E viruses, rotavirus, and sapovirus). DHS influent and effluent were collected weekly or biweekly for 337 days, and concentrations of viral genomes were determined by microfluidic quantitative PCR. Aichivirus, norovirus GI and GII, enterovirus, and sapovirus were frequently detected in DHS influent, and the log 10 reduction (LR) of these viruses ranged from 1.5 to 3.7. The LR values for aichivirus and norovirus GII were also calculated using a Bayesian estimation model, and the average LR (±standard deviation) values for aichivirus and norovirus GII were estimated to be 1.4 (±1.5) and 1.8 (±2.5), respectively. Quantitative microbial risk assessment was conducted to calculate a threshold reduction level for norovirus GII that would be required for the use of DHS effluent for agricultural irrigation, and it was found that LRs of 2.6 and 3.7 for norovirus GII in the DHS effluent were required in order to not exceed the tolerable burden of disease at 10 -4 and 10 -6 disability-adjusted life years loss per person per year, respectively, for 95% of the exposed population during wastewater reuse for irrigation. Copyright © 2016 Elsevier Ltd. All rights reserved.

Inactivation of Salmonella enterica and Listeria monocytogenes in cantaloupe puree by high hydrostatic pressure with/without added ascorbic acid.

PubMed

Mukhopadhyay, Sudarsan; Sokorai, Kimberly; Ukuku, Dike; Fan, Xuetong; Juneja, Vijay; Sites, Joseph; Cassidy, Jennifer

2016-10-17

The objective of this research was to evaluate and develop a method for inactivation of Salmonella enterica and Listeria monocytogenes in cantaloupe puree (CP) by high hydrostatic pressure (HHP). Cantaloupe being the most netted varieties of melons presents a greater risk of pathogen transmission. Freshly prepared CP with or without 0.1% ascorbic acid (AA) was inoculated with a bacterial cocktail composed of a three serotype mixture of S. enterica (S. Poona, S. Newport H1275 and S. Stanley H0558) and a mixture of three strains of L. monocytogenes (Scott A, 43256 and 51742) to a population of ca. 10(8)CFU/g. Double sealed and double bagged inoculated CP (ca. 5g) were pressure treated at 300, 400 and 500MPa at 8°C and 15°C for 5min. Data indicated increased inactivation of both Salmonella and Listeria spp. with higher pressure. Log reduction for CP at 300MPa, 8°C for 5min was 2.4±0.2 and 1.6±0.5logCFU/g for Salmonella and Listeria, respectively. Survivability of the pathogens was significantly compromised at 400MPa and 8°C, inactivating 4.5±0.3logCFU/g of Salmonella and 3.0±0.4logCFU/g of Listeria spp. Complete inactivation of the pathogens in the puree (log reduction >6.7logCFU/g), with or without AA, was achieved when the pressure was further increased to 500MPa, except that for Listeria containing no AA at 8°C. Listeria presented higher resistance to pressure treatment compared to Salmonella spp. Initial temperatures (8 and 15°C) had no significant influence on Salmonella log reductions. Log reduction of pathogens increased but not significantly with increase of temperature. AA did not show any significant antimicrobial activity. Viable counts were about 0.2-0.4logCFU/g less in presence of 0.1% AA. These data validate that HHP can be used as an effective method for decontamination of cantaloupe puree. Published by Elsevier B.V.

Reduction of Escherichia coli O157:H7 and Salmonella on laboratory-inoculated alfalfa seed with commercial citrus-related products.

PubMed

Fett, William F; Cooke, Peter H

2003-07-01

Alfalfa sprouts contaminated with the bacterial pathogens Escherichia coli O157:H7 and Salmonella have been the source of numerous outbreaks of foodborne illness in the United States and in other countries. The seed used for sprouting appears to be the primary source of these pathogens. The aim of this study was to determine whether the efficacy of commercial citrus-related products for sanitizing sprouting seed is similar to that of high levels of chlorine. Five products (Citrex, Pangermex, Citricidal, Citrobio, and Environné) were tested at concentrations of up to 20,000 ppm in sterile tap water and compared with buffered chlorine (at 16,000 ppm). Alfalfa seeds were inoculated with four-strain cocktails of Salmonella and E. coli O157:H7 to give final initial concentrations of ca. 9.0 and 7.0 CFU/g, respectively. Treatments (10 min) with Citrex, Pangermex, and Citricidal at 20,000 ppm and chlorine at 16,000 ppm produced similar log reductions for alfalfa seed inoculated with four-strain cocktails of E. coli O157:H7 and Salmonella (3.42 to 3.46 log CFU/g and 3.56 to 3.74 log CFU/g, respectively), and all four treatments were significantly (P<0.05) more effective than the control treatment (a buffer wash). Citrobio at 20,000 ppm was as effective as the other three products and chlorine against Salmonella but not against E. coli O157:H7. Environné was not more effective (producing reductions of 2.2 to 2.9 log CFU/g) than the control treatment (which produced reductions of 2.1 to 2.3 log CFU/g) against either pathogen. None of the treatments reduced seed germination. In vitro assays, as well as transmission electron microscopy, confirmed the antibacterial nature of the products that were effective against the two pathogens and indicated that they were bactericidal. When used at 20,000 ppm, the effective citrus-related products may be viable alternatives to chlorine for the sanitization of sprouting seed pending regulatory approval.

Detailed analysis of metagenome datasets obtained from biogas-producing microbial communities residing in biogas reactors does not indicate the presence of putative pathogenic microorganisms

PubMed Central

2013-01-01

Background In recent years biogas plants in Germany have been supposed to be involved in amplification and dissemination of pathogenic bacteria causing severe infections in humans and animals. In particular, biogas plants are discussed to contribute to the spreading of Escherichia coli infections in humans or chronic botulism in cattle caused by Clostridium botulinum. Metagenome datasets of microbial communities from an agricultural biogas plant as well as from anaerobic lab-scale digesters operating at different temperatures and conditions were analyzed for the presence of putative pathogenic bacteria and virulence determinants by various bioinformatic approaches. Results All datasets featured a low abundance of reads that were taxonomically assigned to the genus Escherichia or further selected genera comprising pathogenic species. Higher numbers of reads were taxonomically assigned to the genus Clostridium. However, only very few sequences were predicted to originate from pathogenic clostridial species. Moreover, mapping of metagenome reads to complete genome sequences of selected pathogenic bacteria revealed that not the pathogenic species itself, but only species that are more or less related to pathogenic ones are present in the fermentation samples analyzed. Likewise, known virulence determinants could hardly be detected. Only a marginal number of reads showed similarity to sequences described in the Microbial Virulence Database MvirDB such as those encoding protein toxins, virulence proteins or antibiotic resistance determinants. Conclusions Findings of this first study of metagenomic sequence reads of biogas producing microbial communities suggest that the risk of dissemination of pathogenic bacteria by application of digestates from biogas fermentations as fertilizers is low, because obtained results do not indicate the presence of putative pathogenic microorganisms in the samples analyzed. PMID:23557021

Microbial antagonism as a potential solution for controlling selected root pathogens of crops

NASA Astrophysics Data System (ADS)

Cooper, Sarah; Agnew, Linda; Pereg, Lily

2016-04-01

Root pathogens of crops can cause large reduction in yield, however, there is a limited range of effective methods to control such pathogens. Soilborne pathogens that infect roots often need to survive in the rhizosphere, where there is high competition from other organisms. In such hot spots of microbial activity and growth, supported by root exudates, microbes have evolved antagonistic mechanisms that give them competitive advantages in winning the limited resources. Among these mechanisms is antibiosis, with production of some significant antifungal compounds including, antibiotics, volatile organic compounds, hydrogen cyanide and lytic enzymes. Some of these mechanisms may suppress disease through controlling the growth of root pathogens. In this project we isolated various fungi and bacteria that suppress the growth of cotton pathogens in vitro. The pathogen-suppressive microbes were isolated from cotton production soils that are under different management strategies, with and without the use of organic amendments. The potential of pathogen-suppressing microbes for controlling the black root rot disease, caused by the soilborne pathogen Thielaviopsis basicola, was confirmed using soil assays. We identified isolates with potential use as inoculant for cotton production in Australia. Having isolated a diverse group of antagonistic microbes enhances the probability that some would survive well in the soil and provide an alternative approach to address the problem of root disease affecting agricultural crops.

Reduction of Bacterial Pathogens and Potential Surrogates on the Surface of Almonds Using High-Intensity 405-Nanometer Light.

PubMed

Lacombe, Alison; Niemira, Brendan A; Sites, Joseph; Boyd, Glenn; Gurtler, Joshua B; Tyrell, Breanna; Fleck, Melissa

2016-11-01

The disinfecting properties of high-intensity monochromatic blue light (MBL) were investigated against Escherichia coli O157:H7, Salmonella , and nonpathogenic bacteria inoculated onto the surface of almonds. MBL was generated from an array of narrow-band 405-nm light-emitting diodes. Almonds were inoculated with higher or lower levels (8 or 5 CFU/g) of pathogenic E. coli O157:H7 and Salmonella , as well as nonpathogenic E. coli K-12 and an avirulent strain of Salmonella Typhimurium, for evaluation as potential surrogates for their respective pathogens. Inoculated almonds were treated with MBL for 0, 1, 2, 4, 6, 8, and 10 min at a working distance of 7 cm. Simultaneous to treatment, cooling air was directed onto the almonds at a rate of 4 ft 3 /min (1.89 ×10 -3 m 3 /s), sourced through a container of dry ice. An infrared camera was used to monitor the temperature readings after each run. For E. coli K-12, reductions of up to 1.85 or 1.63 log CFU/g were seen for higher and lower inoculum levels, respectively; reductions up to 2.44 and 1.44 log CFU/g were seen for E. coli O157:H7 (higher and lower inoculation levels, respectively). Attenuated Salmonella was reduced by up to 0.54 and 0.97 log CFU/g, whereas pathogenic Salmonella was reduced by up to 0.70 and 0.55 log CFU/g (higher and lower inoculation levels, respectively). Inoculation level did not significantly impact minimum effective treatment times, which ranged from 1 to 4 min. Temperatures remained below ambient throughout treatment, indicating that MBL is a nonthermal antimicrobial process. The nonpathogenic strains of E. coli and Salmonella each responded to MBL in a comparable manner to their pathogenic counterparts. These results suggest that these nonpathogenic strains may be useful in experiments with MBL in which a surrogate is required, and that MBL warrants further investigation as a potential antimicrobial treatment for low-moisture foods.

Simple model of epidemics with pathogen mutation.

PubMed

Girvan, Michelle; Callaway, Duncan S; Newman, M E J; Strogatz, Steven H

2002-03-01

We study how the interplay between the memory immune response and pathogen mutation affects epidemic dynamics in two related models. The first explicitly models pathogen mutation and individual memory immune responses, with contacted individuals becoming infected only if they are exposed to strains that are significantly different from other strains in their memory repertoire. The second model is a reduction of the first to a system of difference equations. In this case, individuals spend a fixed amount of time in a generalized immune class. In both models, we observe four fundamentally different types of behavior, depending on parameters: (1) pathogen extinction due to lack of contact between individuals; (2) endemic infection; (3) periodic epidemic outbreaks; and (4) one or more outbreaks followed by extinction of the epidemic due to extremely low minima in the oscillations. We analyze both models to determine the location of each transition. Our main result is that pathogens in highly connected populations must mutate rapidly in order to remain viable.

Swine and Poultry Pathogens: the Complete Genome Sequences of Two Strains of Mycoplasma hyopneumoniae and a Strain of Mycoplasma synoviae†

PubMed Central

Vasconcelos, Ana Tereza R.; Ferreira, Henrique B.; Bizarro, Cristiano V.; Bonatto, Sandro L.; Carvalho, Marcos O.; Pinto, Paulo M.; Almeida, Darcy F.; Almeida, Luiz G. P.; Almeida, Rosana; Alves-Filho, Leonardo; Assunção, Enedina N.; Azevedo, Vasco A. C.; Bogo, Maurício R.; Brigido, Marcelo M.; Brocchi, Marcelo; Burity, Helio A.; Camargo, Anamaria A.; Camargo, Sandro S.; Carepo, Marta S.; Carraro, Dirce M.; de Mattos Cascardo, Júlio C.; Castro, Luiza A.; Cavalcanti, Gisele; Chemale, Gustavo; Collevatti, Rosane G.; Cunha, Cristina W.; Dallagiovanna, Bruno; Dambrós, Bibiana P.; Dellagostin, Odir A.; Falcão, Clarissa; Fantinatti-Garboggini, Fabiana; Felipe, Maria S. S.; Fiorentin, Laurimar; Franco, Gloria R.; Freitas, Nara S. A.; Frías, Diego; Grangeiro, Thalles B.; Grisard, Edmundo C.; Guimarães, Claudia T.; Hungria, Mariangela; Jardim, Sílvia N.; Krieger, Marco A.; Laurino, Jomar P.; Lima, Lucymara F. A.; Lopes, Maryellen I.; Loreto, Élgion L. S.; Madeira, Humberto M. F.; Manfio, Gilson P.; Maranhão, Andrea Q.; Martinkovics, Christyanne T.; Medeiros, Sílvia R. B.; Moreira, Miguel A. M.; Neiva, Márcia; Ramalho-Neto, Cicero E.; Nicolás, Marisa F.; Oliveira, Sergio C.; Paixão, Roger F. C.; Pedrosa, Fábio O.; Pena, Sérgio D. J.; Pereira, Maristela; Pereira-Ferrari, Lilian; Piffer, Itamar; Pinto, Luciano S.; Potrich, Deise P.; Salim, Anna C. M.; Santos, Fabrício R.; Schmitt, Renata; Schneider, Maria P. C.; Schrank, Augusto; Schrank, Irene S.; Schuck, Adriana F.; Seuanez, Hector N.; Silva, Denise W.; Silva, Rosane; Silva, Sérgio C.; Soares, Célia M. A.; Souza, Kelly R. L.; Souza, Rangel C.; Staats, Charley C.; Steffens, Maria B. R.; Teixeira, Santuza M. R.; Urmenyi, Turan P.; Vainstein, Marilene H.; Zuccherato, Luciana W.; Simpson, Andrew J. G.; Zaha, Arnaldo

2005-01-01

This work reports the results of analyses of three complete mycoplasma genomes, a pathogenic (7448) and a nonpathogenic (J) strain of the swine pathogen Mycoplasma hyopneumoniae and a strain of the avian pathogen Mycoplasma synoviae; the genome sizes of the three strains were 920,079 bp, 897,405 bp, and 799,476 bp, respectively. These genomes were compared with other sequenced mycoplasma genomes reported in the literature to examine several aspects of mycoplasma evolution. Strain-specific regions, including integrative and conjugal elements, and genome rearrangements and alterations in adhesin sequences were observed in the M. hyopneumoniae strains, and all of these were potentially related to pathogenicity. Genomic comparisons revealed that reduction in genome size implied loss of redundant metabolic pathways, with maintenance of alternative routes in different species. Horizontal gene transfer was consistently observed between M. synoviae and Mycoplasma gallisepticum. Our analyses indicated a likely transfer event of hemagglutinin-coding DNA sequences from M. gallisepticum to M. synoviae. PMID:16077101

Development of a risk-based index for source water protection planning, which supports the reduction of pathogens from agricultural activity entering water resources.

PubMed

Goss, Michael; Richards, Charlene

2008-06-01

Source water protection planning (SWPP) is an approach to prevent contamination of ground and surface water in watersheds where these resources may be abstracted for drinking or used for recreation. For SWPP the hazards within a watershed that could contribute to water contamination are identified together with the pathways that link them to the water resource. In rural areas, farms are significant potential sources of pathogens. A risk-based index can be used to support the assessment of the potential for contamination following guidelines on safety and operational efficacy of processes and practices developed as beneficial approaches to agricultural land management. Evaluation of the health risk for a target population requires knowledge of the strength of the hazard with respect to the pathogen load (massxconcentration). Manure handling and on-site wastewater treatment systems form the most important hazards, and both can comprise confined and unconfined source elements. There is also a need to understand the modification of pathogen numbers (attenuation) together with characteristics of the established pathways (surface or subsurface), which allow the movement of the contaminant species from a source to a receptor (water source). Many practices for manure management have not been fully evaluated for their impact on pathogen survival and transport in the environment. A key component is the identification of potential pathways of contaminant transport. This requires the development of a suitable digital elevation model of the watershed for surface movement and information on local groundwater aquifer systems for subsurface flows. Both require detailed soils and geological information. The pathways to surface and groundwater resources can then be identified. Details of land management, farm management practices (including animal and manure management) and agronomic practices have to be obtained, possibly from questionnaires completed by each producer within the watershed. To confirm that potential pathways are active requires some microbial source tracking. One possibility is to identify the molecular types of Escherichia coli present in each hazard on a farm. An essential part of any such index is the identification of mitigation strategies and practices that can reduce the magnitude of the hazard or block open pathways.

How much reduction of virus is needed for recycled water: A continuous changing need for assessment?

PubMed

Gerba, Charles P; Betancourt, Walter Q; Kitajima, Masaaki

2017-01-01

To ensure the safety of wastewater reuse for irrigation of food crops and drinking water pathogenic viruses must be reduced to levels that pose no significant risk. To achieve this goal minimum reduction of viruses by treatment trains have been suggested. For use of edible crops a 6-log reduction and for production of potable drinking water a 12-log reduction has been suggested. These reductions were based on assuming infective virus concentrations of 10 5 to 10 6 per liter. Recent application of molecular methods suggests that some pathogenic viruses may be occurring in concentrations of 10 7 to 10 9 per liter. Factors influencing these levels include the development of molecular methods for virus detection, emergence of newly recognized viruses, decrease in per capita water use due to conservation measures, and outbreaks. Since neither cell culture nor molecular methods can assess all the potentially infectious virus in wastewater conservative estimates should be used to assess the virus load in untreated wastewater. This review indicates that an additional 2- to 3-log reduction of viruses above current recommendations may be needed to ensure the safety of recycled water. Information is needed on peak loading of viruses. In addition, more virus groups need to be quantified using better methods of virus quantification, including more accurate methods for measuring viral infectivity in order to better quantify risks from viruses in recycled water. Copyright © 2016 Elsevier Ltd. All rights reserved.

Application of ozonated dry ice (ALIGAL™ Blue Ice) for packaging and transport in the food industry.

PubMed

Fratamico, Pina M; Juneja, Vijay; Annous, Bassam A; Rasanayagam, Vasuhi; Sundar, M; Braithwaite, David; Fisher, Steven

2012-05-01

Dry ice is used by meat and poultry processors for temperature reduction during processing and for temperature maintenance during transportation. ALIGAL™ Blue Ice (ABI), which combines the antimicrobial effect of ozone (O(3)) along with the high cooling capacity of dry ice, was investigated for its effect on bacterial reduction in air, in liquid, and on food and glass surfaces. Through proprietary means, O(3) was introduced to produce dry ice pellets to a concentration of 20 parts per million (ppm) by total weight. The ABI sublimation rate was similar to that of dry ice pellets under identical conditions, and ABI was able to hold the O(3) concentration throughout the normal shelf life of the product. Challenge studies were performed using different microorganisms, including E. coli, Campylobacter jejuni, Salmonella, and Listeria, that are critical to food safety. ABI showed significant (P < 0.05) microbial reduction during bioaerosol contamination (up to 5-log reduction of E. coli and Listeria), on chicken breast (approximately 1.3-log reduction of C. jejuni), on contact surfaces (approximately 3.9 log reduction of C. jejuni), and in liquid (2-log reduction of C. jejuni). Considering the stability of O(3), ease of use, and antimicrobial efficacy against foodborne pathogens, our results suggest that ABI is a better alternative, especially for meat and poultry processors, as compared to dry ice. Further, ABI can potentially serve as an additional processing hurdle to guard against pathogens during processing, transportation, distribution, and/or storage. © 2012 Institute of Food Technologists®

Monte Carlo simulation of the risk of contamination of apples with Escherichia coli O157:H7.

PubMed

Duffy, Siobain; Schaffner, Donald W

2002-10-25

Quantitative descriptions of the frequency and extent of contamination of apple cider with pathogenic bacteria were obtained using literature data and computer simulation. Probability distributions were chosen to describe the risk of apple contamination by each suspected pathway. Tree-picked apples may be contaminated by birds infected with Escherichia coli O157:H7 when orchards were located near a sewage source (ocean or landfill). Dropped apples could become contaminated from either infected animal droppings or from contaminated manure if used as fertilizer. A risk assessment model was created in Analytica. The results of worst-case simulations revealed that 6-9 log CFU E. coli O157:H7 might be found on a harvest of 1000 dropped apples, while 3-4 log CFU contamination could be present on 1000 tree-picked apples. This model confirms that practices such as using dropped apples and using animal waste as fertilizer increase risk in the production of apple cider, and that pasteurization may not eliminate all contamination in juice from heavily contaminated fruit. Recently published FDA regulations for juices requiring a 5-log CFU/ml reduction of pathogenic bacteria in fresh juices should be a fail-safe measure for apples harvested in all but the worst-case scenarios.

Thermal inactivation of H5N2 high-pathogenicity avian influenza virus in dried egg white with 7.5% moisture.

PubMed

Thomas, Colleen; Swayne, David E

2009-09-01

High-pathogenicity avian influenza viruses (HPAIV) cause severe systemic disease with high mortality in chickens. Isolation of HPAIV from the internal contents of chicken eggs has been reported, and this is cause for concern because HPAIV can be spread by movement of poultry products during marketing and trade activity. This study presents thermal inactivation data for the HPAIV strain A/chicken/PA/1370/83 (H5N2) (PA/83) in dried egg white with a moisture content (7.5%) similar to that found in commercially available spray-dried egg white products. The 95% upper confidence limits for D-values calculated from linear regression of the survival curves at 54.4, 60.0, 65.5, and 71.1 degrees C were 475.4, 192.2, 141.0, and 50.1 min, respectively. The line equation y = [0.05494 x degrees C] + 5.5693 (root mean square error = 0.0711) was obtained by linear regression of experimental D-values versus temperature. Conservative predictions based on the thermal inactivation data suggest that standard industry pasteurization protocols would be very effective for HPAIV inactivation in dried egg white. For example, these calculations predict that a 7-log reduction would take only 2.6 days at 54.4 degrees C.

Predicting Salmonella populations from biological, chemical, and physical indicators in Florida surface waters.

PubMed

McEgan, Rachel; Mootian, Gabriel; Goodridge, Lawrence D; Schaffner, Donald W; Danyluk, Michelle D

2013-07-01

Coliforms, Escherichia coli, and various physicochemical water characteristics have been suggested as indicators of microbial water quality or index organisms for pathogen populations. The relationship between the presence and/or concentration of Salmonella and biological, physical, or chemical indicators in Central Florida surface water samples over 12 consecutive months was explored. Samples were taken monthly for 12 months from 18 locations throughout Central Florida (n = 202). Air and water temperature, pH, oxidation-reduction potential (ORP), turbidity, and conductivity were measured. Weather data were obtained from nearby weather stations. Aerobic plate counts and most probable numbers (MPN) for Salmonella, E. coli, and coliforms were performed. Weak linear relationships existed between biological indicators (E. coli/coliforms) and Salmonella levels (R(2) < 0.1) and between physicochemical indicators and Salmonella levels (R(2) < 0.1). The average rainfall (previous day, week, and month) before sampling did not correlate well with bacterial levels. Logistic regression analysis showed that E. coli concentration can predict the probability of enumerating selected Salmonella levels. The lack of good correlations between biological indicators and Salmonella levels and between physicochemical indicators and Salmonella levels shows that the relationship between pathogens and indicators is complex. However, Escherichia coli provides a reasonable way to predict Salmonella levels in Central Florida surface water through logistic regression.

Predicting Salmonella Populations from Biological, Chemical, and Physical Indicators in Florida Surface Waters

PubMed Central

McEgan, Rachel; Mootian, Gabriel; Goodridge, Lawrence D.; Schaffner, Donald W.

2013-01-01

Coliforms, Escherichia coli, and various physicochemical water characteristics have been suggested as indicators of microbial water quality or index organisms for pathogen populations. The relationship between the presence and/or concentration of Salmonella and biological, physical, or chemical indicators in Central Florida surface water samples over 12 consecutive months was explored. Samples were taken monthly for 12 months from 18 locations throughout Central Florida (n = 202). Air and water temperature, pH, oxidation-reduction potential (ORP), turbidity, and conductivity were measured. Weather data were obtained from nearby weather stations. Aerobic plate counts and most probable numbers (MPN) for Salmonella, E. coli, and coliforms were performed. Weak linear relationships existed between biological indicators (E. coli/coliforms) and Salmonella levels (R2 < 0.1) and between physicochemical indicators and Salmonella levels (R2 < 0.1). The average rainfall (previous day, week, and month) before sampling did not correlate well with bacterial levels. Logistic regression analysis showed that E. coli concentration can predict the probability of enumerating selected Salmonella levels. The lack of good correlations between biological indicators and Salmonella levels and between physicochemical indicators and Salmonella levels shows that the relationship between pathogens and indicators is complex. However, Escherichia coli provides a reasonable way to predict Salmonella levels in Central Florida surface water through logistic regression. PMID:23624476

[Effects of "host factor" bile on adaptability and virulence of Vibrios, foodborne potential pathogenic agents].

PubMed

Di Pietro, A; Picerno, I; Visalli, G; Chirico, C; Scoglio, M E

2004-01-01

In order to improve the knowledge of host/pathogenic agent interaction and to obtain a more careful estimation of risk related to ingestion of food contaminated by Vibrio spp., the effects of bile extracts have been studied. The growth of one V. fluvialis, two V. alginolyticus, and three V. parahaemolyticus strains, isolated from mollusks and crustaceans, has been determined to evaluate their adaptability to intestinal environment. Moreover, the expression of virulence factors responsible for the colonization, as bacterial "swarming mobility", biofilm production, adherence on epithelial cells and hydrophobicity, has been evaluated. Using a bile concentration of 1.5%, all examined strains showed a constant inhibitory effect, quite moderate in the first growth phases. Bile increased the "swarming mobility" and biofilm production; also the adherence was favored, but only after adaptation and during the early logarithmic phase. The decreased hydrophobicity could explain the reduction of adherence during the stationary phase. Studying the phenotypic expression of virulence factors in "minor vibrios" in the presence of bile, it was possible to extend the knowledge about their pathogenetic mechanisms owing to the ingestion of contaminated food. That permits a more careful estimation of risk related to the contamination, considering the high frequency of isolation of these species in some seafood.

Rainfall and temperatures changes have confounding impacts on Phytophthora cinnamomi occurrence risk in the southwestern USA under climate change scenarios.

PubMed

Thompson, Sally E; Levin, Simon; Rodriguez-Iturbe, Ignacio

2014-04-01

Global change will simultaneously impact many aspects of climate, with the potential to exacerbate the risks posed by plant pathogens to agriculture and the natural environment; yet, most studies that explore climate impacts on plant pathogen ranges consider individual climatic factors separately. In this study, we adopt a stochastic modeling approach to address multiple pathways by which climate can constrain the range of the generalist plant pathogen Phytophthora cinnamomi (Pc): through changing winter soil temperatures affecting pathogen survival; spring soil temperatures and thus pathogen metabolic rates; and changing spring soil moisture conditions and thus pathogen growth rates through host root systems. We apply this model to the southwestern USA for contemporary and plausible future climate scenarios and evaluate the changes in the potential range of Pc. The results indicate that the plausible range of this pathogen in the southwestern USA extends over approximately 200,000 km(2) under contemporary conditions. While warming temperatures as projected by the IPCC A2 and B1 emissions scenarios greatly expand the range over which the pathogen can survive winter, projected reductions in spring rainfall reduce its feasible habitat, leading to spatially complex patterns of changing risk. The study demonstrates that temperature and rainfall changes associated with possible climate futures in the southwestern USA have confounding impacts on the range of Pc, suggesting that projections of future pathogen dynamics and ranges should account for multiple pathways of climate-pathogen interaction. © 2014 John Wiley & Sons Ltd.

Use of organic acids to inactivate Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes on organic fresh apples and lettuce.

PubMed

Park, Sang-Hyun; Choi, Mi-Ran; Park, Jeong-Woong; Park, Ki-Hwan; Chung, Myung-Sub; Ryu, Sangryeol; Kang, Dong-Hyun

2011-08-01

This study was undertaken to investigate the antimicrobial effect of organic acids against Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes on whole red organic apples and lettuce. Several studies have been conducted to evaluate organic acids as sanitizers. However, no studies have compared antimicrobial effects of various organic acids on organic fresh produce, including evaluation of color changes of produce. Apples and lettuce were inoculated with a cocktail of 3 strains each of 3 foodborne pathogens provided above and treated with 1% and 2% organic acids (propionic, acetic, lactic, malic, and citric acid) for 0, 0.5, 1, 5, and 10 min. With increasing treatment time and acid concentration, organic acid treatments showed significant reduction compared to the control treatment (distilled water), and differences in antimicrobial effects between organic acids were observed. After 10 min of treatment with 1% and 2% organic acids in apples, propionic (0.92 to 2.75 log reduction), acetic (0.52 to 2.78 log reduction), lactic (1.69 to >3.42 log reduction), malic (1.48 to >3.42 log reduction), and citric acid (1.52 to >3.42 log reduction) exhibited significant (P < 0.05) antibacterial effects against 3 foodborne pathogens compared to the control treatment. In lettuce, propionic (0.93 to 1.52 log reduction), acetic (1.13 to 1.74 log reduction), lactic (1.87 to 2.54 log reduction), malic (2.32 to 2.98 log reduction), and citric acid (1.85 to 2.86 log reduction) showed significant (P < 0.05) effects compared to the control treatment. Changes in sample color subjected to organic acids treatment were not significant during storage. It is suggested that organic acids have a potential as sanitizers for organic fresh produce. These data may help the organic produce industry provide safe fresh produce for consumers. © 2011 Institute of Food Technologists®

Hygienic effects and gas production of plastic bio-digesters under tropical conditions.

PubMed

Yen-Phi, Vo Thi; Clemens, Joachim; Rechenburg, Andrea; Vinneras, Björn; Lenssen, Christina; Kistemann, Thomas

2009-12-01

Plastic plug-flow bio-digesters have been promoted as a good option for improved treatment of manure and wastewater in developing countries although minimal information has been published on their hygienic status. This bench-scale study replicates bio-digester conditions to evaluate the reduction of pathogen and indicator microorganisms at three different hydraulic retention times (HRT) in the anaerobic treatment of pig manures at 30 degrees C for 50 days. Results showed that physicochemical values differed between HRTs. Gas production efficiency was better for longer HRTS. The accumulated sludge at the reactor's base increased with longer HRT. Phages and bacteria examined were reduced, but none was completely eliminated. Log10 reduction of bacteria ranged from 0.54 to 2.47. Phages ranged from 1.60 to 3.42. The reduction of organisms at HRT = 30 days was about one log10 unit higher than HRT = 15 days and about two log10 units higher than HRT = 3 days. The results indicate that the reduction of tested organisms increases with HRT. However the hygienic quality of the liquid effluent does not meet required quality values for surface and irrigation water. Longer HRTs are recommended to increase gas yield and achieve higher pathogen reduction. More barriers should be applied while handling bio-digester outputs to minimise risks to environmental and human health.

[Innovative technology and blood safety].

PubMed

Begue, S; Morel, P; Djoudi, R

2016-11-01

If technological innovations are not enough alone to improve blood safety, their contributions for several decades in blood transfusion are major. The improvement of blood donation (new apheresis devices, RFID) or blood components (additive solutions, pathogen reduction technology, automated processing of platelets concentrates) or manufacturing process of these products (by automated processing of whole blood), all these steps where technological innovations were implemented, lead us to better traceability, more efficient processes, quality improvement of blood products and therefore increased blood safety for blood donors and patients. If we are on the threshold of a great change with the progress of pathogen reduction technology (for whole blood and red blood cells), we hope to see production of ex vivo red blood cells or platelets who are real and who open new conceptual paths on blood safety. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

Biocontrol of the wheat pathogen Zymoseptoria tritici using cyclic lipopeptides from Bacillus subtilis.

PubMed

Mejri, Samara; Siah, Ali; Coutte, François; Magnin-Robert, Maryline; Randoux, Béatrice; Tisserant, Benoit; Krier, François; Jacques, Philippe; Reignault, Philippe; Halama, Patrice

2017-06-21

Innovation toward ecofriendly plant protection products compatible with sustainable agriculture and healthy food is today strongly encouraged. Here, we assessed the biocontrol activity of three cyclic lipopeptides from Bacillus subtilis (mycosubtilin, M; surfactin, S; fengycin, F) and two mixtures (M + S and M + S + F) on wheat against Zymoseptoria tritici, the main pathogen on this crop. Foliar application of these biomolecules at a 100-mg L -1 concentration on the wheat cultivars Dinosor and Alixan, 2 days before fungal inoculation, provided significant reductions of disease severity. The best protection levels were recorded with the M-containing formulations (up to 82% disease reduction with M + S on Dinosor), while S and F treatments resulted in lower but significant disease reductions. In vitro and in planta investigations revealed that M-based formulations inhibit fungal growth, with half-maximal inhibitory concentrations of 1.4 mg L -1 for both M and M + S and 4.5 mg L -1 for M + S + F, thus revealing that the observed efficacy of these products may rely mainly on antifungal property. By contrast, S and F had no direct activity on the pathogen, hence suggesting that these lipopeptides act on wheat against Z. tritici as resistance inducers rather than as biofungicides. This study highlighted the efficacy of several lipopeptides from B. subtilis to biocontrol Z. tritici through likely distinct and biomolecule-dependent modes of action.

Effect of different GRAS compounds in the control of apples blue mould.

PubMed

Venditti, T; Cubaiu, L; Ladu, G; D'Hallewin, G

2013-01-01

The most important pathogen for apples is Penicillium expansum that is the causal organism of blue mould. Postharvest losses are controlled with chemical fungicides such as TBZ but a growing concern for human health and a greater awareness for environmental conservation have multiplied the studies on new ecological technologies. In the search of new environment and consumer friendly technologies that can reduce toxic residues, the use of GRAS compounds represent a valid alternative to the use of synthetic postharvest fungicides. The aims of the present work were: (1) To evaluate the effectiveness of different GRAS compounds in the control of P. expansum in artificially inoculated fruit; (2) To assess the capability of injured and treated fruit with GRAS compounds, used alone or combined, to heal the wounds in order to resist to infection. Fruit was injured with a steel rod and after 1 hour was (1) Inoculated with the pathogen and after 24 hours treated or (2) Treated and after 24 hours inoculated. Treatments were performed with the following compounds: sodium bicarbonate (SBC), boric acid (BA) and calcium chloride (CC) by using a 1% solution for all of them. After 9 or 14 days fruit lesion diameters were assessed. In the trial (1) the combined treatment with BA and SBC was the most effective reducing the lesion diameter by 86.5% with respect to untreated fruit, after 9 days from infection. A good pathogen control was also obtained with BA used alone or combined with CC. When the treatment was performed before infection the best results were achieved with the combination of SBC and CC, with 87% of reduction of the lesion diameter. The addition of CC also reduced the lesion if combined with BA (66.8%). These preliminary results showed that GRAS compounds can be effective in reducing blue mold by a direct effect on the pathogen, and by modulating fruit responses enhancing host resistance.

Potassium permanganate cleansing is an effective sanitary method for the reduction of bacterial bioload on raw Coriandrum sativum.

PubMed

Subramanya, Supram Hosuru; Pai, Vasudha; Bairy, Indira; Nayak, Niranjan; Gokhale, Shishir; Sathian, Brijesh

2018-02-13

Raw vegetables including flowers, leaves, stems, and roots are important carriers of food borne pathogens. We evaluated the bacteriological contamination of unwashed coriander leaves, and effectiveness of cleansing with 0.1% potassium permanganate solution as decontamination method. Significant bacterial contamination including pathogens like Salmonella species and Aeromonas species were isolated from unwashed coriander leaves. Decontamination with 0.1% potassium permanganate was found to be more effective than three steps wash with sterile water.

Efficacy of titanium dioxide nanoparticle spray to disinfect mobile phones used by endodontist: A bacteriological study.

PubMed

Palaniswamy, Udayakumar; Habeeb, Aisha; Mohsin, Mohammed

2018-01-01

The objectives of this study were to determine the bacterial contamination of the mobile phones that were used by the endodontist in comparison with the general dentist and also to determine the usefulness of titanium dioxide nanospray (TiO 2 NS) in mobile phone decontamination. Samples from the 60 mobile phones were taken using moist sterile swabs before, 10 min, and 1 week after the use of TiO 2 NS. Before collection of the swabs, the participants' informed consent was obtained. Samples obtained were cultured on blood agar to identify bacterial isolates. All 60 mobile phone cultures were found to be culture positive. There was a significant reduction in the mean number of colony-forming units after decontamination with TiO 2 NS ( P < 0.001). The results from this study showed that the mobile phones may act as an important source of nosocomial pathogens and TiO 2 NS would be an effective decontaminant. Therefore, it is important for dental professionals to practice routine mobile phone disinfection protocol to reduce the chances of occurrence of nosocomial infections.

Application of the laser to the study of pathogenic fungi.

PubMed

Thibaut, M

1979-05-15

Laser microanalysis has been applied to the study of pathogenic fungi. Such a method allows chemical information to be obtained and permits the detection of 74 elements in the periodic system of Mendeleev from lithium (3) to uranium (92).

Changes in oral microflora after full-mouth tooth extraction: a prospective cohort study.

PubMed

de Waal, Yvonne C M; Winkel, Edwin G; Raangs, Gerwin C; van der Vusse, Marleen L; Rossen, John W A; van Winkelhoff, Arie Jan

2014-10-01

The aim of the study was to evaluate the effect of full-mouth tooth extraction on the oral microflora, with emphasis on the presence and load of Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis. Adult patients (n = 30), with moderate to advanced periodontitis and scheduled for full-mouth tooth extraction, were consecutively selected. Prior to and 1 and 3 months after full-mouth tooth extraction saliva, tongue, buccal and gingival mucosa and subgingival plaque/prosthesis samples were obtained. Aerobic and anaerobic culture techniques and quantitative real-time polymerase chain reaction (qPCR) were employed for the detection of oral pathogens. Full-mouth tooth extraction resulted in reduction below detection level of A. actinomycetemcomitans and P. gingivalis in 15 of 16 and 8 of 16 previously positive patients using culture techniques and qPCR, respectively. Those patients remaining qPCR positive showed a significant reduction in load of these bacteria. Full-mouth tooth extraction significantly changes the oral microflora. These changes include reduction of A. actinomycetemcomitans and P. gingivalis, frequently to levels below detection threshold. In some patients, A. actinomycetemcomitans and P. gingivalis can persist in the edentulous oral cavity up to 3 months after full-mouth tooth extraction. © 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Case-Control Pilot Study on Acute Diarrheal Disease in a Geographically Defined Pediatric Population in a Middle Income Country

PubMed Central

Farfán-García, Ana E.; Zhang, Chengxian; Imdad, Aamer; Arias-Guerrero, Monica Y.; Sánchez-Alvarez, Nayibe T.; Shah, Rikhil; Iqbal, Junaid; Tamborski, Maria E.

2017-01-01

Introduction Acute diarrheal disease (ADD) is a common cause of morbidity and mortality in children under 5 years of age. Understanding of the etiology of ADD is lacking in most low and middle income countries because reference laboratories detect limited number of pathogens. The objective of this study was to determine the feasibility to conduct a comprehensive case-control study to survey diarrheal pathogens among children with and without moderate-to-severe ADD. Materials and Methods Microbiology and molecular-based techniques were used to detect viral, bacterial, and parasitic enteropathogens. The study was conducted in Bucaramanga, Colombia, after Institutional Review Board approval was obtained. Results Ninety children less than 5 years of age were recruited after a written informed consent was obtained from parents or guardians. Forty-five subjects served as cases with ADD and 45 as controls. Thirty-six subjects out of 90 (40.0%) were positive for at least one enteropathogen, that is, 20 (44.4%) cases and 16 (35.5%) controls. Conclusions The three most common enteric pathogens were enteroaggregative E. coli (10.0%), Norovirus (6.7%), and Salmonella spp. (5.6%). The E. coli pathogens were 18.8% of all infections making them the most frequent pathogens. Half of ADD cases were negative for any pathogens. PMID:28855919

A novel pathogenicity gene is required in the rice blast fungus to suppress the basal defenses of the host.

PubMed

Chi, Myoung-Hwan; Park, Sook-Young; Kim, Soonok; Lee, Yong-Hwan

2009-04-01

For successful colonization and further reproduction in host plants, pathogens need to overcome the innate defenses of the plant. We demonstrate that a novel pathogenicity gene, DES1, in Magnaporthe oryzae regulates counter-defenses against host basal resistance. The DES1 gene was identified by screening for pathogenicity-defective mutants in a T-DNA insertional mutant library. Bioinformatic analysis revealed that this gene encodes a serine-rich protein that has unknown biochemical properties, and its homologs are strictly conserved in filamentous Ascomycetes. Targeted gene deletion of DES1 had no apparent effect on developmental morphogenesis, including vegetative growth, conidial germination, appressorium formation, and appressorium-mediated penetration. Conidial size of the mutant became smaller than that of the wild type, but the mutant displayed no defects on cell wall integrity. The Deltades1 mutant was hypersensitive to exogenous oxidative stress and the activity and transcription level of extracellular enzymes including peroxidases and laccases were severely decreased in the mutant. In addition, ferrous ion leakage was observed in the Deltades1 mutant. In the interaction with a susceptible rice cultivar, rice cells inoculated with the Deltades1 mutant exhibited strong defense responses accompanied by brown granules in primary infected cells, the accumulation of reactive oxygen species (ROS), the generation of autofluorescent materials, and PR gene induction in neighboring tissues. The Deltades1 mutant displayed a significant reduction in infectious hyphal extension, which caused a decrease in pathogenicity. Notably, the suppression of ROS generation by treatment with diphenyleneiodonium (DPI), an inhibitor of NADPH oxidases, resulted in a significant reduction in the defense responses in plant tissues challenged with the Deltades1 mutant. Furthermore, the Deltades1 mutant recovered its normal infectious growth in DPI-treated plant tissues. These results suggest that DES1 functions as a novel pathogenicity gene that regulates the activity of fungal proteins, compromising ROS-mediated plant defense.

Liquid based formulations of bacteriophages for the management of waterborne bacterial pathogens in water microcosms.

PubMed

Ahiwale, Sangeeta; Tagunde, Sujata; Khopkar, Sushama; Karni, Mrudula; Gajbhiye, Milind; Kapadnis, Balasaheb

2013-11-01

Water resources are contaminated by life-threatening multidrug resistant pathogenic bacteria. Unfortunately, these pathogenic bacteria do not respond to the traditional water purification methods. Therefore, there is a need of environmentally friendly strategies to overcome the problems associated with the antimicrobial resistant bacterial pathogens. In the present study, highly potent lytic phages against multidrug-resistant Salmonella enterica serovar Paratyphi B, Pseudomonas aeruginosa and Klebsiella pneumoniae were isolated from the Pavana river water. They belonged to the Podoviridae and Siphoviridae families. These phages were purified and enriched in the laboratory. Monovalent formulations of phiSPB, BVPaP-3 and KPP phages were prepared in three different liquids viz., phage broth, saline and distilled water. The phages were stable for almost 8-10 months in the phage broth at 4 degrees C. The stability of the phages in saline and distilled water was 5-6 months at 4 degrees C. All of the phages were stable only for 4-6 months in the phage broth at 30 degrees C. The monovalent phage formulation of psiSPB was applied at MOI < 1, as disinfectant against an exponential and stationary phase cells of Salmonella enterica serovar Paratyphi B in various water microcosms. The results indicated that there was almost 80 % reduction in the log phase cells of Salmonella serovar Paratyphi B in 24 h. In stationary phase cells, the reduction was comparatively less within same period. At the same time, there was concomitant increase in the phage population by 80% in all the microcosms indicating that psiSPB phage is highly potent in killing pathogen in water. Results strongly support that the formulation of psiSPB in the phage broth in monovalent form could be used as an effective biological disinfectant for preventing transmission of water-borne bacterial pathogens, including antimicrobial resistant ones.

Efficiency of different air filter types for pig facilities at laboratory scale

PubMed Central

Wenke, Cindy; Pospiech, Janina; Reutter, Tobias; Truyen, Uwe

2017-01-01

Air filtration has been shown to be efficient in reducing pathogen burden in circulating air. We determined at laboratory scale the retention efficiency of different air filter types either composed of a prefilter (EU class G4) and a secondary fiberglass filter (EU class F9) or consisting of a filter mat (EU class M6 and F8-9). Four filter prototypes were tested for their capability to remove aerosol containing equine arteritis virus (EAV), porcine reproductive and respiratory syndrome virus (PRRSV), bovine enterovirus 1 (BEV), Actinobacillus pleuropneumoniae (APP), and Staphylococcus (S.) aureus from air. Depending on the filter prototype and utilisation, the airflow was set at 1,800 m3/h (combination of upstream prefilter and fiberglass filter) or 80 m3/h (filter mat). The pathogens were aerosolized and their concentration was determined in front of and behind the filter by culture or quantitative real-time RT-PCR. Furthermore, survival of the pathogens over time in the filter material was determined. Bacteria were most efficiently filtered with a reduction rate of up to 99.9% depending on the filter used. An approximately 98% reduction was achieved for the viruses tested. Viability or infectivity of APP or PRRSV in the filter material decreased below the detection limit after 4 h and 24 h, respectively, whereas S. aureus was still culturable after 4 weeks. Our results demonstrate that pathogens can efficiently be reduced by air filtration. Consequently, air filtration combined with other strict biosecurity measures markedly reduces the risk of introducing airborne transmitted pathogens to animal facilities. In addition, air filtration might be useful in reducing bioaerosols within a pig barn, hence improving respiratory health of pigs. PMID:29028843

Pseudomonas fluorescens CHA0 maintains carbon delivery to Fusarium graminearum-infected roots and prevents reduction in biomass of barley shoots through systemic interactions

PubMed Central

Henkes, Gunnar J.; Jousset, Alexandre; Bonkowski, Michael; Thorpe, Michael R.; Scheu, Stefan; Lanoue, Arnaud; Schurr, Ulrich; Röse, Ursula S. R.

2011-01-01

Soil bacteria such as pseudomonads may reduce pathogen pressure for plants, both by activating plant defence mechanisms and by inhibiting pathogens directly due to the production of antibiotics. These effects are hard to distinguish under field conditions, impairing estimations of their relative contributions to plant health. A split-root system was set up with barley to quantify systemic and local effects of pre-inoculation with Pseudomonas fluorescens on the subsequent infection process by the fungal pathogen Fusarium graminearum. One root half was inoculated with F. graminearum in combination with P. fluorescens strain CHA0 or its isogenic antibiotic-deficient mutant CHA19. Bacteria were inoculated either together with the fungal pathogen or in separate halves of the root system to separate local and systemic effects. The short-term plant response to fungal infection was followed by using the short-lived isotopic tracer 11CO2 to track the delivery of recent photoassimilates to each root half. In the absence of bacteria, fungal infection diverted carbon from the shoot to healthy roots, rather than to infected roots, although the overall partitioning from the shoot to the entire root system was not modified. Both local and systemic pre-inoculation with P. fluorescens CHA0 prevented the diversion of carbon as well as preventing a reduction in plant biomass in response to F. graminearum infection, whereas the non-antibiotic-producing mutant CHA19 lacked this ability. The results suggest that the activation of plant defences is a central feature of biocontrol bacteria which may even surpass the effects of direct pathogen inhibition. PMID:21561952

Pathogen reduction and blood transfusion safety in Africa: strengths, limitations and challenges of implementation in low-resource settings.

PubMed

Ware, A D; Jacquot, C; Tobian, A A R; Gehrie, E A; Ness, P M; Bloch, E M

2018-01-01

Transfusion-transmitted infection risk remains an enduring challenge to blood safety in Africa. A high background incidence and prevalence of the major transfusion-transmitted infections (TTIs), dependence on high-risk donors to meet demand, suboptimal testing and quality assurance collectively contribute to the increased risk. With few exceptions, donor testing is confined to serological evaluation of human immunodeficiency virus (HIV), hepatitis B and C (HBV and HCV) and syphilis. Barriers to implementation of broader molecular methods include cost, limited infrastructure and lack of technical expertise. Pathogen reduction (PR), a term used to describe a variety of methods (e.g. solvent detergent treatment or photochemical activation) that may be applied to blood following collection, offers the means to diminish the infectious potential of multiple pathogens simultaneously. This is effective against different classes of pathogen, including the major TTIs where laboratory screening is already implemented (e.g. HIV, HBV and HCV) as well pathogens that are widely endemic yet remain unaddressed (e.g. malaria, bacterial contamination). We sought to review the available and emerging PR techniques and their potential application to resource-constrained parts of Africa, focusing on the advantages and disadvantages of such technologies. PR has been slow to be adopted even in high-income countries, primarily given the high costs of use. Logistical considerations, particularly in low-resourced parts of Africa, also raise concerns about practicality. Nonetheless, PR offers a rational, innovative strategy to contend with TTIs; technologies in development may well present a viable complement or even alternative to targeted screening in the future. © 2017 International Society of Blood Transfusion.

Efficiency of different air filter types for pig facilities at laboratory scale.

PubMed

Wenke, Cindy; Pospiech, Janina; Reutter, Tobias; Truyen, Uwe; Speck, Stephanie

2017-01-01

Air filtration has been shown to be efficient in reducing pathogen burden in circulating air. We determined at laboratory scale the retention efficiency of different air filter types either composed of a prefilter (EU class G4) and a secondary fiberglass filter (EU class F9) or consisting of a filter mat (EU class M6 and F8-9). Four filter prototypes were tested for their capability to remove aerosol containing equine arteritis virus (EAV), porcine reproductive and respiratory syndrome virus (PRRSV), bovine enterovirus 1 (BEV), Actinobacillus pleuropneumoniae (APP), and Staphylococcus (S.) aureus from air. Depending on the filter prototype and utilisation, the airflow was set at 1,800 m3/h (combination of upstream prefilter and fiberglass filter) or 80 m3/h (filter mat). The pathogens were aerosolized and their concentration was determined in front of and behind the filter by culture or quantitative real-time RT-PCR. Furthermore, survival of the pathogens over time in the filter material was determined. Bacteria were most efficiently filtered with a reduction rate of up to 99.9% depending on the filter used. An approximately 98% reduction was achieved for the viruses tested. Viability or infectivity of APP or PRRSV in the filter material decreased below the detection limit after 4 h and 24 h, respectively, whereas S. aureus was still culturable after 4 weeks. Our results demonstrate that pathogens can efficiently be reduced by air filtration. Consequently, air filtration combined with other strict biosecurity measures markedly reduces the risk of introducing airborne transmitted pathogens to animal facilities. In addition, air filtration might be useful in reducing bioaerosols within a pig barn, hence improving respiratory health of pigs.

Quantification of pathogen inactivation efficacy by free chlorine disinfection of drinking water for QMRA.

PubMed

Petterson, S R; Stenström, T A

2015-09-01

To support the implementation of quantitative microbial risk assessment (QMRA) for managing infectious risks associated with drinking water systems, a simple modeling approach for quantifying Log10 reduction across a free chlorine disinfection contactor was developed. The study was undertaken in three stages: firstly, review of the laboratory studies published in the literature; secondly, development of a conceptual approach to apply the laboratory studies to full-scale conditions; and finally implementation of the calculations for a hypothetical case study system. The developed model explicitly accounted for variability in residence time and pathogen specific chlorine sensitivity. Survival functions were constructed for a range of pathogens relying on the upper bound of the reported data transformed to a common metric. The application of the model within a hypothetical case study demonstrated the importance of accounting for variable residence time in QMRA. While the overall Log10 reduction may appear high, small parcels of water with short residence time can compromise the overall performance of the barrier. While theoretically simple, the approach presented is of great value for undertaking an initial assessment of a full-scale disinfection contactor based on limited site-specific information.

Kinetics of inactivation of indicator pathogens during thermophilic anaerobic digestion.

PubMed

Popat, Sudeep C; Yates, Marylynn V; Deshusses, Marc A

2010-12-01

Thermophilic anaerobic sludge digestion is a promising process to divert waste to beneficial use, but an important question is the required temperature and holding time to achieve a given degree of pathogen inactivation. In this study, the kinetics of inactivation of Ascaris suum and vaccine strain poliovirus type 1 (PVS-1), selected as indicators for helminth ova and enteric viruses respectively, were determined during anaerobic digestion at temperatures ranging from 51 to 56 °C. Inactivation of both indicator organisms was fast with greater than two log reductions achieved within 2 h for A. suum and three log reductions for PVS-1, suggesting that the current U.S. regulations are largely conservative. The first-order inactivation rate constants k followed Arrhenius relationship with activation energies of 105 and 39 KJ mol(-1) for A. suum and PVS-1, respectively indicating that A. suum was more sensitive to temperature. Although inactivation was fast, the presence of compounds in the sludge that are known to be protective of pathogen inactivation was observed, suggesting that composition-dependent time-temperature relationships are necessary. Copyright © 2010 Elsevier Ltd. All rights reserved.

Effects of combined treatments of irradiation and antimicrobial coatings on reduction of food pathogens in broccoli florets

NASA Astrophysics Data System (ADS)

Takala, P. N.; Salmieri, S.; Vu, K. D.; Lacroix, M.

2011-12-01

The effect of combined treatment of antimicrobial coatings and γ-radiation on reduction of food pathogens such as Listeria monocytogenes, Escherichia coli, and Salmonella Typhimurium was evaluated in broccoli florets. Broccoli florets were inoculated with pathogenic bacteria at 10 6 CFU/g. Inoculated florets were then coated with methylcellulose-based coating containing various mixtures of antimicrobial agents: organic acids (OAs) plus lactic acid bacteria metabolites (LABs), OA plus citrus extract (CE), OA plus CE plus spice mixture (SM), and OA plus rosemary extract (RE). Coated florets were irradiated with various doses (0-3.3 kGy), and microbial analyses were used to calculate the D10 value and radiosensitive relative. The coating containing OA plus CE was the most effective formulation for increasing the sensitization of Escherichia coli by 2.4 times as compared to the control without the antimicrobial coating. For Salmonella Typhimurium, coating containing OA plus LAB was the most effective formulation, increasing radiosensitivity by 2.4 times as well. All antimicrobial coatings had almost the same effect of increasing the sensitivity of Listeria monocytogenes (from 1.31 to 1.45 times) to γ-irradiation.

Impact of antibiotic administration on blood culture positivity at the beginning of sepsis: a prospective clinical cohort study.

PubMed

Scheer, Christian S; Fuchs, Christian; Gründling, Matthias; Vollmer, Marcus; Bast, Juliane; Bohnert, Jürgen A; Zimmermann, Kathrin; Hahnenkamp, Klaus; Rehberg, Sebastian; Kuhn, Sven-Olaf

2018-06-04

Sepsis guidelines recommend obtaining blood cultures before starting anti-infective therapy in patients with sepsis. However, little is known how antibiotic treatment prior to sampling affects bacterial growth. The aim of this study was to compare the results of blood cultures drawn prior to and under antibiotic therapy. Prospective clinical cohort study of septic patients. Adult ICU patients with 2 or 3 blood culture (BC) sets at the beginning of sepsis between 2010 and 2017 were included. Patients with blood culture samplings obtained prior to antibiotic therapy were compared to patients with samplings under antibiotic therapy. Blood culture positivity, defined as microbiological pathogen finding, was compared between the groups. Logistic regression was performed to adjust the impact of different factors with respect to blood culture positivity. In total, 559 patients with 1364 blood culture sets at the beginning of sepsis were analyzed. BC positivity was 50.6% (78/154) among septic patients who did not receive antibiotics and only 27.7% (112/405) in those who were already under antibiotics (P<0.001). Logistic regression revealed antibiotic therapy as an independent factor for less pathogen identification (Odds ratio 0.4; 95%CI 0.3-0.6). Gram-positive pathogens (28.3%(111/392) vs. 11.9%(116/972);P<0.001) and also gram-negative pathogens (16.3%(64/392) vs. 9.3%(90/972);P<0.001) were more frequent in BC sets drawn prior to antibiotic therapy compared to sets under antibiotics. Obtaining blood cultures under antibiotic therapy is associated with a significant loss of pathogen detection. This strongly emphasizes the current recommendation to obtain blood cultures prior to antibiotic administration in patients with sepsis. Copyright © 2018. Published by Elsevier Ltd.

Biological control of postharvest pathogens

USDA-ARS?s Scientific Manuscript database

Postharvest diseases of harvested commodities cause significant reductions in food availability and financial profits. Additionally, regulatory agencies are increasingly restricting or banning the postharvest use of synthetic chemical fungicides. This has increased the need to develop more ecofrie...

The Effect of Resin and Monoterpenes on Spore Germination and Growth in Fusarium circinatum.

PubMed

Slinski, S L; Zakharov, F; Gordon, T R

2015-01-01

Resin obtained from Pinus radiata and five monoterpene components of resin (limonene, α-pinene, β-pinene, camphene, and myrcene) were tested to determine their effects on mycelial growth and germination and survival of spores of Fusarium circinatum, the cause of pitch canker in pine, and F. temperatum, which is interfertile with F. circinatum but not pathogenic to pine. Averaged across all treatments, F. temperatum sustained the greatest reduction in radial growth (16.9±0.02% of control). The greatest reduction in dry weight also occurred in F. temperatum (11.7±0.01% of control), and all isolates of F. circinatum were significantly less affected (P<0.05). Spore germination rates in a saturated atmosphere of monoterpenes were relatively high for all tested isolates but, when placed in direct contact with resin, spore survival was significantly greater for F. circinatum than for F. temperatum. Our results are consistent with the hypothesis that greater tolerance of resin is one factor distinguishing F. circinatum from the nonpathogenic F. temperatum. However, differential tolerance of monoterpene components of resin is not sufficient to explain the observed variation in virulence to pine in F. circinatum.

Effect of Infection by Beauveria bassiana and Metarhizium anisopliae on the Feeding of Uvarovistia zebra

PubMed Central

Mohammadbeigi, A.; Port, G.

2015-01-01

To identify the susceptibility of long-horned grasshoppers to entomopathogenic fungi, the effect of infection with the fungi Beauveria bassiana (Balsamo) Vuillemin (Hypocreales: Clavicipitaceae) and Metarhizium anisopliae (Metchnikoff) Sorokin (Hypocreales: Clavicipitaceae) on food consumption by Uvarovistia zebra (Uvarov) (Orthoptera: Tettigoniidae) was investigated. Preliminary results showed that both fungi had a negative effect on food consumption of the insects. For both fungi a significant reduction of food consumption and faeces production by insects were observed between the highest spore concentration (5 × 106 spores/ml) and other treatments. Compared with control insects, the insects treated with 5 × 106 spores/ml of B. bassiana and M. anisopliae showed 60 and 63% reduction in mean food consumption/insect, respectively. The corrected cumulative percent mortality of the insects treated with the highest concentration of B. bassiana and M. anisopliae were 57.7 and 55.5%, respectively. This was the first account of these entomopathogenic fungi being used against a species from this family, therefore based on the results obtained from this research, it could be said that the fungi have pathogenicity effect on U. zebra as a long-horned grasshopper.

Preliminary research concerning the use of electron accelerators to improve the conservability and to extend the shelf-life of fruits and vegetables

NASA Astrophysics Data System (ADS)

Minea, R.; Oproiu, C.; Pascanu, S.; Matei, C.; Ferdes, O.

1996-06-01

The potential of ionizing radiation treatment for food preservation, shelf-life extension, control of microbial load and reduction of pathogenic microorganism was demonstrated. The irradiations were performed under normal conditions on the Institute of Physics and Technology for Radiation Device's linear electron accelerator, which has the following parameters: 5 μA mean beam current, 6 MeV electron mean energy, pulse period 3.5 μs and dose rates between 100-1500 Gy/min. This research project was aimed at assuring the consumer's acceptance for radiation-treated food and to obtain a significant reduction of food losses. We also propose a promising solution for the radiation processing of some bulk food products at the place of storage, consisting of a mobile electron accelerator. The main characteristics of the mobile electron accelerator are: electron energy 3 to 5 MeV, maximum beam power 5 kW, vertical electron beam; irradiation is possible both with electron beams and with bremsstrahlung. The results of our preliminary research lead to the conclusion that electron-beam irradiation and the use of electron accelerators is a promising solution for food preservation and food safety. Interesting future applications are outlined.

Modeling disinfection of plastic poultry transport cages inoculated with Salmonella enteritids by slightly acidic electrolyzed water using response surface methodology.

PubMed

Zang, Y T; Li, B M; Bing, Sh; Cao, W

2015-09-01

In order to reduce the risk of enteric pathogens transmission in animal farms, the disinfection effectiveness of slightly acidic electrolyzed water (SAEW, pH 5.85 to 6.53) for inactivating Salmonella Enteritidis on the surface of plastic poultry transport cages was evaluated. The coupled effects of the tap water cleaning time (5 to 15 s), SAEW treatment time (20 to 40 s), and available chlorine concentrations (ACCs) of 30 to 70 mg/l on the reductions of S. Enteritidis on chick cages were investigated using a central composite design of the response surface methodology (RSM). The established RS model had a goodness of fit quantified by the parameter R2 (0.971), as well as a lack of fit test (P>0.05). The maximum reduction of 3.12 log10 CFU/cm2 for S. Enteritidis was obtained for the cage treated with tap water cleaning for 15 s followed by SAEW treatment for 40 s at an ACC of 50 mg/l. Results indicate that the established RS model has shown the potential of SAEW in disinfection of bacteria on cages. © 2015 Poultry Science Association Inc.

Disinfection and reduction of organic load of sewage water by electron beam radiation

NASA Astrophysics Data System (ADS)

Maruthi, Y. Avasn; Das, N. Lakshmana; Hossain, Kaizar; Sarma, K. S. S.; Rawat, K. P.; Sabharwal, S.

2011-09-01

The efficacy of electron beam radiation for the disinfection and reduction of organic load of sewage water was assessed with ILU-6 Accelerator at Radiation Technology Development Division of the Bhabha Atomic Research Centre, Mumbai India. The current problem on environmental health in relation to water pollution insists for the safe disposal of sewage water. In general, sewage water comprises heterogeneous organic based chemicals as well as pathogens. EB treatment of the wastewater has found to be very effective in reducing the pathogens as well as organic load. EB dose of 1.5 kGy was sufficient for complete elimination of total coli forms. The experimental results elucidated the reduction of biological oxygen demand—BOD (35 and 51.7%) in both inlet and outlet sewage samples. Similarly reduction of chemical oxygen demand—COD was observed (37.54 and 52.32%) in both sewage samples with respect to increase in irradiation doses (0.45-6 kGy). The present study demonstrated the potential of ionizing radiation for disinfection of sewage and to increase the water quality of the wastewater by decreasing BOD and COD. So, the irradiation sewage water can find its application either in agriculture for irrigation, in industry for cooling purpose and some selected domestic purposes.

5-Nitroimidazole-derived Schiff bases and their copper(II) complexes exhibit potent antimicrobial activity against pathogenic anaerobic bacteria.

PubMed

Oliveira, Alexandre A; Oliveira, Ana P A; Franco, Lucas L; Ferencs, Micael O; Ferreira, João F G; Bachi, Sofia M P S; Speziali, Nivaldo L; Farias, Luiz M; Magalhães, Paula P; Beraldo, Heloisa

2018-05-07

In the present work a family of novel secnidazole-derived Schiff base compounds and their copper(II) complexes were synthesized. The antimicrobial activities of the compounds were evaluated against clinically important anaerobic bacterial strains. The compounds exhibited in vitro antibacterial activity against Bacteroides fragilis, Bacteroides thetaiotaomicron, Bacteroides vulgatus, Bacteroides ovatus, Parabacteroides distasonis and Fusubacterium nucleatum pathogenic anaerobic bacteria. Upon coordination to copper(II) the antibacterial activity significantly increased in several cases. Some derivatives were even more active than the antimicrobial drugs secnidazole and metronidazole. Therefore, the compounds under study are suitable for in vivo evaluation and the microorganisms should be classified as susceptible to them. Electrochemical studies on the reduction of the nitro group revealed that the compounds show comparable reduction potentials, which are in the same range of the bio-reducible drugs secnidazole and benznidazole. The nitro group reduction potential is more favorable for the copper(II) complexes than for the starting ligands. Hence, the antimicrobial activities of the compounds under study might in part be related to intracellular bio-reduction activation. Considering the increasing resistance rates of anaerobic bacteria against a wide range of antimicrobial drugs, the present work constitutes an important contribution to the development of new antibacterial drug candidates.

Technologies for reducing sludge production in wastewater treatment plants: State of the art.

PubMed

Wang, Qilin; Wei, Wei; Gong, Yanyan; Yu, Qiming; Li, Qin; Sun, Jing; Yuan, Zhiguo

2017-06-01

This review presents the state-of-the-art sludge reduction technologies applied in both wastewater and sludge treatment lines. They include chemical, mechanical, thermal, electrical treatment, addition of chemical un-coupler, and predation of protozoa/metazoa in wastewater treatment line, and physical, chemical and biological pretreatment in sludge treatment line. Emphasis was put on their effect on sludge reduction performance, with 10% sludge reduction to zero sludge production in wastewater treatment line and enhanced TS (total solids) or volatile solids removal of 5-40% in sludge treatment line. Free nitrous acid (FNA) technology seems good in wastewater treatment line but it is only under the lab-scale trial. In sludge treatment line, thermal, ultrasonic (<4400kJ/kg TS), FNA pretreatment and temperature-phased anaerobic digestion (TPAD) are promising if pathogen inactivation is not a concern. However, thermal pretreatment and TPAD are superior to other pretreatment technologies when pathogen inactivation is required. The new wastewater treatment processes including SANI®, high-rate activated sludge coupled autotrophic nitrogen removal and anaerobic membrane bioreactor coupled autotrophic nitrogen removal also have a great potential to reduce sludge production. In the future, an effort should be put on the effect of sludge reduction technologies on the removal of organic micropollutants and heavy metals. Copyright © 2017 Elsevier B.V. All rights reserved.

Pathogen dynamics during invasion and establishment of white-nose syndrome explain mechanisms of host persistence.

PubMed

Frick, Winifred F; Cheng, Tina L; Langwig, Kate E; Hoyt, Joseph R; Janicki, Amanda F; Parise, Katy L; Foster, Jeffrey T; Kilpatrick, A Marm

2017-03-01

Disease dynamics during pathogen invasion and establishment determine the impacts of disease on host populations and determine the mechanisms of host persistence. Temporal progression of prevalence and infection intensity illustrate whether tolerance, resistance, reduced transmission, or demographic compensation allow initially declining populations to persist. We measured infection dynamics of the fungal pathogen Pseudogymnoascus destructans that causes white-nose syndrome in bats by estimating pathogen prevalence and load in seven bat species at 167 hibernacula over a decade as the pathogen invaded, became established, and some host populations stabilized. Fungal loads increased rapidly and prevalence rose to nearly 100% at most sites within 2 yr of invasion in six of seven species. Prevalence and loads did not decline over time despite huge reductions in colony sizes, likely due to an extensive environmental reservoir. However, there was substantial variation in fungal load among sites with persisting colonies, suggesting that both tolerance and resistance developed at different sites in the same species. In contrast, one species disappeared from hibernacula within 3 yr of pathogen invasion. Variable host responses to pathogen invasion require different management strategies to prevent disease-induced extinction and to facilitate evolution of tolerance or resistance in persisting populations. © 2016 by the Ecological Society of America.

Impact of pathogen-directed antimicrobial therapy for ventilator-associated pneumonia in trauma patients on charges and recurrence.

PubMed

Sharpe, John P; Magnotti, Louis J; Weinberg, Jordan A; Swanson, Joseph M; Wood, G Christopher; Fabian, Timothy C; Croce, Martin A

2015-04-01

Ventilator-associated pneumonia (VAP) represents one of the driving forces behind antibiotic use in the ICU. In a previous study, we established a defined algorithm for treatment of hospital-acquired VAP dictated by the causative pathogen. The purpose of the current study was to evaluate the impact of this algorithm for hospital-acquired VAP on recurrence and charges in trauma patients. Patients with VAP secondary to MRSA, Acinetobacter baumannii, Pseudomonas aeruginosa, Stenotrophomonas maltophilia, or Enterobacteriaceae during 5 years subsequent to the previous study were evaluated. All VAP were diagnosed using quantitative cultures of the bronchoalveolar lavage effluent. Duration of antimicrobial therapy was dictated by the causative pathogen. If microbiologic resolution, defined as <10(3) colony-forming units/mL, was achieved, therapy was stopped by day 10. The remainder received 14 days of therapy. Recurrence was defined as >10(5) colony-forming units/mL on subsequent bronchoalveolar lavage performed within 2 weeks after completion of appropriate therapy. Five hundred and twenty-nine VAP episodes were identified in 381 patients. Overall recurrence was unchanged compared with the previous study (1.5% vs 2%; p = 0.3). There was a decrease in the number of bronchoalveolar lavages performed per patient compared with the previous study (1.6 vs 2.3; p = 0.24) and a reduction of 4.8 antibiotic days per VAP episode compared with the previous study. Both changes resulted in a cumulative reduction of $3,535.04 per patient, for a savings of $1.35 million during the study period. Hospital-acquired VAP can be managed effectively by a defined course of therapy dictated by the causative pathogen. Adherence to an established algorithm simplified the management of VAP and contributed to a cumulative reduction in patient charges without impacting recurrence. Copyright © 2015 American College of Surgeons. Published by Elsevier Inc. All rights reserved.

Antibacterial and efflux pump inhibitors of thymol and carvacrol against food-borne pathogens.

PubMed

Miladi, Hanene; Zmantar, Tarek; Chaabouni, Yassine; Fedhila, Kais; Bakhrouf, Amina; Mahdouani, Kacem; Chaieb, Kamel

2016-10-01

In this study thymol (THY) and carvacrol (CAR), two monoterpenic phenol produced by various aromatic plants, was tested for their antibacterial and efflux pump inhibitors potencies against a panel of clinical and foodborne pathogenes. Our results demonstrated a substantial susceptibility of the tested bacteria toward THY and CAR. Especially, THY displayed a strong inhibitory activity (MIC's values ranged from 32 to 64 μg/mL) against the majority of the tested strains compared to CAR. Moreover, a significant reduction in MIC's of TET and benzalkonium chloride (QAC) were noticed when tested in combinations with THY and CAR. Their synergic effect was more significant in the case of THY which resulted a reduction of MIC's values of TET (2-8 fold) and QAC (2-8 fold). We noted also that THY and CAR inhibited the ethidium bromide (EtBr) cell efflux in a concentration-dependent manner. The rate of EtBr accumulation in food-borne pathogen was enhanced with THY and CAR (0, 250 and 500 μg/mL). The lowest concentration causing 50% of EtBr efflux inhibition (IC 50) was noticed in Salmonella enteritidis (1129) at 150 μg/mL of THY and 190 μg/mL of CAR respectively. These findings indicate that THY and CAR may serve as potential sources of efflux pump inhibitor in food-borne pathogens. Copyright © 2016 Elsevier Ltd. All rights reserved.

Predators indirectly reduce the prevalence of an insect-vectored plant pathogen independent of predator diversity.

PubMed

Long, Elizabeth Y; Finke, Deborah L

2015-04-01

A widely cited benefit of predator diversity is greater suppression of insect herbivores, with corresponding increases in plant biomass. In the context of a vector-borne pathogen system, predator species richness may also influence plant disease risk via the direct effects of predators on the abundance and behavior of herbivores that also act as pathogen vectors. Using an assemblage of generalist insect predators, we examined the relationship between predator species richness and the prevalence of the aphid-vectored cereal yellow dwarf virus in wheat. We found that increasing predator richness enhanced suppression of the vector population and that pathogen prevalence was reduced when predators were present, but the reduction in prevalence was independent of predator species richness. To determine the mechanism(s) by which predator species richness contributes to vector suppression, but not pathogen prevalence, we evaluated vector movement and host plant occupancy in response to predator treatments. We found that pathogen prevalence was unrelated to vector suppression because host plant occupancy by vectors did not vary as a function of vector abundance. However, the presence of predators reduced pathogen prevalence because predators stimulated greater plant-to-plant movement by vectors, which likely diminished vector feeding time and reduced the transmission efficiency of this persistent pathogen. We conclude that community structure (i.e., the presence of predators), but not predator diversity, is a potential factor influencing local plant infection by this insect-vectored pathogen.

The application of food safety interventions in primary production of beef and lamb: a review.

PubMed

Adam, K; Brülisauer, F

2010-07-31

The production of safe red meat depends on effective control of pathogenic microorganisms at all stages of the "farm-to-fork" chain. Eight microorganisms have been selected as the focus of the PathogenCombat project: Shiga toxin producing Escherichia coli (STEC), Mycobacterium avium subspecies paratuberculosis (Map), Listeria monocytogenes, Campylobacter jejuni, Penicillium nordicum, invasive variants of Saccharomyces cerevisiae, hepatitis E virus and tick borne encephalitis virus. The need and potential for coordinated control of the selected food-borne pathogens by on-farm interventions is assessed using a decision tree and a review of the relevant scientific literature. Control measures to reduce the carriage of these pathogens in ruminants prior to slaughter are reviewed with reference to the current regulations and guidelines relating to the primary production. From the eight pathogens investigated, two (STEC and Map), are likely to be effectively controlled by interventions at farm level and the applicable interventions are described and discussed. Ruminants are the main reservoir for these two pathogens; hence a reduction of carriage in livestock should directly reduce human exposure through the consumption of beef and lamb. Copyright 2009 Elsevier B.V. All rights reserved.

Pathogen prevalence and abundance in honey bee colonies involved in almond pollination.

PubMed

Cavigli, Ian; Daughenbaugh, Katie F; Martin, Madison; Lerch, Michael; Banner, Katie; Garcia, Emma; Brutscher, Laura M; Flenniken, Michelle L

Honey bees are important pollinators of agricultural crops. Since 2006, US beekeepers have experienced high annual honey bee colony losses, which may be attributed to multiple abiotic and biotic factors, including pathogens. However, the relative importance of these factors has not been fully elucidated. To identify the most prevalent pathogens and investigate the relationship between colony strength and health, we assessed pathogen occurrence, prevalence, and abundance in Western US honey bee colonies involved in almond pollination. The most prevalent pathogens were Black queen cell virus (BQCV), Lake Sinai virus 2 (LSV2), Sacbrood virus (SBV), Nosema ceranae , and trypanosomatids. Our results indicated that pathogen prevalence and abundance were associated with both sampling date and beekeeping operation, that prevalence was highest in honey bee samples obtained immediately after almond pollination, and that weak colonies had a greater mean pathogen prevalence than strong colonies.

Occurrence and antimicrobial resistance of pathogenic Escherichia coli and Salmonella spp. in retail raw table eggs sold for human consumption in Enugu state, Nigeria

PubMed Central

Okorie-Kanu, O. Josephine; Ezenduka, E. Vivienne; Okorie-Kanu, C. Onwuchokwe; Ugwu, L. Chinweokwu; Nnamani, U. John

2016-01-01

Aim: This study was conducted to investigate the occurrence of pathogenic Escherichia coli and Salmonella species in retail raw table eggs sold for human consumption in Enugu State and to determine the resistance of these pathogens to antimicrobials commonly used in human and veterinary practices in Nigeria. Materials and Methods: A total of 340 raw table eggs comprising 68 composite samples (5 eggs per composite sample) were collected from five selected farms (13 composite samples from the farms) and 10 retail outlets (55 composite samples from the retail outlets) in the study area over a period of 4-month (March-June, 2014). The eggs were screened for pathogenic E. coli and Salmonella species following standard procedures within 24 h of sample collection. Isolates obtained were subjected to in-vitro antimicrobial susceptibility test with 15 commonly used antimicrobials using the disk diffusion method. Results: About 37 (54.4%) and 7 (10.3%) of the 68 composite samples were positive for pathogenic E. coli and Salmonella species, respectively. The shells showed significantly higher (p<0.05) contaminations than the contents for both microorganisms. The eggs from the farms showed higher contamination with pathogenic E. coli than eggs from the retail outlets while the reverse was the case for Salmonella species even though they were not significant (p>0.05). The organisms obtained showed a multiple drug resistance. They were completely resistant to nitrofurantoin, sulfamethoxazole/trimethoprim, penicillin G and oxacillin. In addition to these, Salmonella spp. also showed 100% resistance to tetracycline. The pathogenic E. coli isolates obtained were 100% susceptible to gentamicin, neomycin, ciprofloxacin, and amoxicillin-clavulanic acid while Salmonella spp. showed 100% susceptibility to erythromycin, neomycin, and rifampicin. Both organisms showed varying degrees of resistance to streptomycin, amoxicillin, vancomycin, and doxycycline. Conclusion: From the results of the study, it can be concluded that the raw table eggs marketed for human consumption in Enugu State, Nigeria is contaminated with pathogenic E. coli and Salmonella species that showed multiple drug resistance to antimicrobial agents commonly used in veterinary and human practice. PMID:27956787

Occurrence and antimicrobial resistance of pathogenic Escherichia coli and Salmonella spp. in retail raw table eggs sold for human consumption in Enugu state, Nigeria.

PubMed

Okorie-Kanu, O Josephine; Ezenduka, E Vivienne; Okorie-Kanu, C Onwuchokwe; Ugwu, L Chinweokwu; Nnamani, U John

2016-11-01

This study was conducted to investigate the occurrence of pathogenic Escherichia coli and Salmonella species in retail raw table eggs sold for human consumption in Enugu State and to determine the resistance of these pathogens to antimicrobials commonly used in human and veterinary practices in Nigeria. A total of 340 raw table eggs comprising 68 composite samples (5 eggs per composite sample) were collected from five selected farms (13 composite samples from the farms) and 10 retail outlets (55 composite samples from the retail outlets) in the study area over a period of 4-month (March-June, 2014). The eggs were screened for pathogenic E. coli and Salmonella species following standard procedures within 24 h of sample collection. Isolates obtained were subjected to in-vitro antimicrobial susceptibility test with 15 commonly used antimicrobials using the disk diffusion method. About 37 (54.4%) and 7 (10.3%) of the 68 composite samples were positive for pathogenic E. coli and Salmonella species, respectively. The shells showed significantly higher (p<0.05) contaminations than the contents for both microorganisms. The eggs from the farms showed higher contamination with pathogenic E. coli than eggs from the retail outlets while the reverse was the case for Salmonella species even though they were not significant (p>0.05). The organisms obtained showed a multiple drug resistance. They were completely resistant to nitrofurantoin, sulfamethoxazole/trimethoprim, penicillin G and oxacillin. In addition to these, Salmonella spp. also showed 100% resistance to tetracycline. The pathogenic E. coli isolates obtained were 100% susceptible to gentamicin, neomycin, ciprofloxacin, and amoxicillin-clavulanic acid while Salmonella spp. showed 100% susceptibility to erythromycin, neomycin, and rifampicin. Both organisms showed varying degrees of resistance to streptomycin, amoxicillin, vancomycin, and doxycycline. From the results of the study, it can be concluded that the raw table eggs marketed for human consumption in Enugu State, Nigeria is contaminated with pathogenic E. coli and Salmonella species that showed multiple drug resistance to antimicrobial agents commonly used in veterinary and human practice.

Autoheated thermophilic aerobic digestion

DOE Office of Scientific and Technical Information (OSTI.GOV)

Deeny, K.; Hahn, H.; Leonhard, D.

1991-10-01

Autothermal thermophilic aerobic digestion (ATAD) is first and foremost a digestion process, the primary purpose of which is to decompose a portion of the waste organic solids generated from wastewater treatment. As a result of the high operating temperature, digestion is expected to occur within a short time period (6 days) and accomplish a high degree of pathogen reduction. ATAD systems are two-stage aerobic digestion processes that operate under thermophilic temperature conditions (40 to 80C) without supplemental heat. Like composting, the systems rely on the conservation of heat released during digestion itself to attain and sustain the desired operating temperature.more » Typical ATAD systems operate at 55C and may reach temperatures of 60 to 65C in the second-stage reactor. Perhaps because of the high operating temperature, this process has been referred to as Liquid Composting.' Major advantages associated with thermophilic operation include high biological reaction rates and a substantial degree of pathogen reduction.« less

Chemical generation of nitric oxide in the mouth from the enterosalivary circulation of dietary nitrate.

PubMed

Duncan, C; Dougall, H; Johnston, P; Green, S; Brogan, R; Leifert, C; Smith, L; Golden, M; Benjamin, N

1995-06-01

High concentrations of nitrite present in saliva (derived from dietary nitrate) may, upon acidification, generate nitrogen oxides in the stomach in sufficient amounts to provide protection from swallowed pathogens. We now show that, in the rat, reduction of nitrate to nitrite is confined to a specialized area on the posterior surface of the tongue, which is heavily colonized by bacteria, and that nitrate reduction is absent in germ-free rats. We also show that in humans increased salivary nitrite production resulting from nitrate intake enhances oral nitric oxide production. We propose that the salivary generation of nitrite is accomplished by a symbiotic relationship involving nitrate-reducing bacteria on the tongue surface, which is designed to provide host defence against microbial pathogens in the mouth and lower gut. These results provide further evidence for beneficial effects of dietary nitrate.

Human Immunity and the Design of Multi-Component, Single Target Vaccines

PubMed Central

Saul, Allan; Fay, Michael P.

2007-01-01

Background Inclusion of multiple immunogens to target a single organism is a strategy being pursued for many experimental vaccines, especially where it is difficult to generate a strongly protective response from a single immunogen. Although there are many human vaccines that contain multiple defined immunogens, in almost every case each component targets a different pathogen. As a consequence, there is little practical experience for deciding where the increased complexity of vaccines with multiple defined immunogens vaccines targeting single pathogens will be justifiable. Methodology/Principal Findings A mathematical model, with immunogenicity parameters derived from a database of human responses to established vaccines, was used to predict the increase in the efficacy and the proportion of the population protected resulting from addition of further immunogens. The gains depended on the relative protection and the range of responses in the population to each immunogen and also to the correlation of the responses between immunogens. In most scenarios modeled, the gain in overall efficacy obtained by adding more immunogens was comparable to gains obtained from a single immunogen through the use of better formulations or adjuvants. Multi-component single target vaccines were more effective at decreasing the proportion of poor responders than increasing the overall efficacy of the vaccine in a population. Conclusions/Significance Inclusion of limited number of antigens in a vaccine aimed at targeting a single organism will increase efficacy, but the gains are relatively modest and for a practical vaccine there are constraints that are likely to limit multi-component single target vaccines to a small number of key antigens. The model predicts that this type of vaccine will be most useful where the critical issue is the reduction in proportion of poor responders. PMID:17786221

Molecular methods to assess Listeria monocytogenes route of contamination in a dairy processing plant.

PubMed

Alessandria, Valentina; Rantsiou, Kalliopi; Dolci, Paola; Cocolin, Luca

2010-07-31

In this study we investigated the occurrence of Listeria monocytogenes in a dairy processing plant during two sampling campaigns in 2007 and 2008. Samples represented by semifinished and finished cheeses, swabs from the equipment and brines from the salting step, were subjected to analysis by using traditional and molecular methods, represented mainly by quantitative PCR. Comparing the results obtained by the application of the two approaches used, it became evident how traditional microbiological analysis underestimated the presence of L. monocytogenes in the dairy plant. Especially samples of the brines and the equipment swabs were positive only with qPCR. For some equipment swabs it was possible to detect a load of 10(4)-10(5) cfu/cm(2), while the modified ISO method employed gave negative results both before and after the enrichment step. The evidences collected during the first sampling year, highlighting a heavy contamination of the brines and of the equipment, lead to the implementation of specific actions that decreased the contamination in these samples during the 2008 campaign. However, no reduction in the number of L. monocytogenes positive final products was observed, suggesting that a more strict control is necessary to avoid the presence of the pathogen. All the isolates of L. monocytogenes were able to attach to abiotic surfaces, and, interestingly, considering the results obtained from their molecular characterization it became evident how strains present in the brines, were genetically connected with isolates from the equipment and from the final product, suggesting a clear route of contamination of the pathogen in the dairy plant. This study underlines the necessity to use appropriate analytical tools, such as molecular methods, to fully understand the spread and persistence of L. monocytogenes in food producing companies. Copyright 2010 Elsevier B.V. All rights reserved.

Treatment of whole blood with riboflavin plus ultraviolet light, an alternative to gamma irradiation in the prevention of transfusion-associated graft-versus-host disease?

PubMed

Fast, Loren D; Nevola, Martha; Tavares, Jennifer; Reddy, Heather L; Goodrich, Ray P; Marschner, Susanne

2013-02-01

Exposure of blood products to gamma irradiation is currently the standard of care in the prevention of transfusion-associated graft-versus-host disease (TA-GVHD). Regulatory, technical, and clinical challenges associated with the use of gamma irradiators are driving efforts to develop alternatives. Pathogen reduction methods were initially developed to reduce the risk of microbial transmission by blood components. Through modifications of nucleic acids, these technologies interfere with the replication of both pathogens and white blood cells (WBCs). To date, systems for pathogen and WBC inactivation of products containing red blood cells are less well established than those for platelets and plasma. In this study, the in vitro and in vivo function of WBCs present in whole blood after exposure to riboflavin plus ultraviolet light (Rb-UV) was examined and compared to responses of WBCs obtained from untreated or gamma-irradiated blood by measuring proliferation, cytokine production, activation, and antigen presentation and xenogeneic (X-)GVHD responses in an in vivo mouse model. In vitro studies demonstrated that treatment of whole blood with Rb-UV was as effective as gamma irradiation in preventing WBC proliferation, but was more effective in preventing antigen presentation, cytokine production, and T-cell activation. Consistent with in vitro findings, treatment with Rb-UV was as effective as gamma irradiation in preventing X-GVHD, a mouse model for TA-GVHD. The ability to effectively inactivate WBCs in fresh whole blood using Rb-UV, prior to separation into components, provides the transfusion medicine community with a potential alternative to gamma irradiation. © 2012 American Association of Blood Banks.

Pyramiding, alternating or mixing: comparative performances of deployment strategies of nematode resistance genes to promote plant resistance efficiency and durability.

PubMed

Djian-Caporalino, Caroline; Palloix, Alain; Fazari, Ariane; Marteu, Nathalie; Barbary, Arnaud; Abad, Pierre; Sage-Palloix, Anne-Marie; Mateille, Thierry; Risso, Sabine; Lanza, Roger; Taussig, Catherine; Castagnone-Sereno, Philippe

2014-02-22

Resistant cultivars are key elements for pathogen control and pesticide reduction, but their repeated use may lead to the emergence of virulent pathogen populations, able to overcome the resistance. Increased research efforts, mainly based on theoretical studies, explore spatio-temporal deployment strategies of resistance genes in order to maximize their durability. We evaluated experimentally three of these strategies to control root-knot nematodes: cultivar mixtures, alternating and pyramiding resistance genes, under controlled and field conditions over a 3-years period, assessing the efficiency and the durability of resistance in a protected crop rotation system with pepper as summer crop and lettuce as winter crop. The choice of the resistance gene and the genetic background in which it is introgressed, affected the frequency of resistance breakdown. The pyramiding of two different resistance genes in one genotype suppressed the emergence of virulent isolates. Alternating different resistance genes in rotation was also efficient to decrease virulent populations in fields due to the specificity of the virulence and the trapping effect of resistant plants. Mixing resistant cultivars together appeared as a less efficient strategy to control nematodes. This work provides experimental evidence that, in a cropping system with seasonal sequences of vegetable species, pyramiding or alternating resistance genes benefit yields in the long-term by increasing the durability of resistant cultivars and improving the long-term control of a soil-borne pest. To our knowledge, this result is the first one obtained for a plant-nematode interaction, which helps demonstrate the general applicability of such strategies for breeding and sustainable management of resistant cultivars against pathogens.

Pyramiding, alternating or mixing: comparative performances of deployment strategies of nematode resistance genes to promote plant resistance efficiency and durability

PubMed Central

2014-01-01

Background Resistant cultivars are key elements for pathogen control and pesticide reduction, but their repeated use may lead to the emergence of virulent pathogen populations, able to overcome the resistance. Increased research efforts, mainly based on theoretical studies, explore spatio-temporal deployment strategies of resistance genes in order to maximize their durability. We evaluated experimentally three of these strategies to control root-knot nematodes: cultivar mixtures, alternating and pyramiding resistance genes, under controlled and field conditions over a 3-years period, assessing the efficiency and the durability of resistance in a protected crop rotation system with pepper as summer crop and lettuce as winter crop. Results The choice of the resistance gene and the genetic background in which it is introgressed, affected the frequency of resistance breakdown. The pyramiding of two different resistance genes in one genotype suppressed the emergence of virulent isolates. Alternating different resistance genes in rotation was also efficient to decrease virulent populations in fields due to the specificity of the virulence and the trapping effect of resistant plants. Mixing resistant cultivars together appeared as a less efficient strategy to control nematodes. Conclusions This work provides experimental evidence that, in a cropping system with seasonal sequences of vegetable species, pyramiding or alternating resistance genes benefit yields in the long-term by increasing the durability of resistant cultivars and improving the long-term control of a soil-borne pest. To our knowledge, this result is the first one obtained for a plant-nematode interaction, which helps demonstrate the general applicability of such strategies for breeding and sustainable management of resistant cultivars against pathogens. PMID:24559060

Potential of Pseudomonas putida PCI2 for the Protection of Tomato Plants Against Fungal Pathogens.

PubMed

Pastor, Nicolás; Masciarelli, Oscar; Fischer, Sonia; Luna, Virginia; Rovera, Marisa

2016-09-01

Tomato is one of the most economically attractive vegetable crops due to its high yields. Diseases cause significant losses in tomato production worldwide. We carried out Polymerase Chain Reaction studies to detect the presence of genes encoding antifungal compounds in the DNA of Pseudomonas putida strain PCI2. We also used liquid chromatography-electrospray tandem mass spectrometry to detect and quantify the production of compounds that increase the resistance of plants to diseases from culture supernatants of PCI2. In addition, we investigated the presence of 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase in PCI2. Finally, PCI2 was used for inoculation of tomato seeds to study its potential biocontrol activity against Fusarium oxysporum MR193. The obtained results showed that no fragments for the encoding genes of hydrogen cyanide, pyoluteorin, 2,4-diacetylphloroglucinol, pyrrolnitrin, or phenazine-1-carboxylic acid were amplified from the DNA of PCI2. On the other hand, PCI2 produced salicylic acid and jasmonic acid in Luria-Bertani medium and grew in a culture medium containing ACC as the sole nitrogen source. We observed a reduction in disease incidence from 53.33 % in the pathogen control to 30 % in tomato plants pre-inoculated with PCI2 as well as increases in shoot and root dry weights in inoculated plants, as compared to the pathogenicity control. This study suggests that inoculation of tomato seeds with P. putida PCI2 increases the resistance of plants to root rot caused by F. oxysporum and that PCI2 produces compounds that may be involved at different levels in increasing such resistance. Thus, PCI2 could represent a non-contaminating management strategy potentially applicable in vegetable crops such as tomato.

Evaluation of the potential of Trichoderma viride in the control of fungal pathogens of Roselle (Hibiscus sabdariffa L.) in vitro.

PubMed

Eslaminejad Parizi, T; Ansaria, Mehdi; Elaminejad, Tahereh

2012-04-01

The potential of Trichoderma viride as a bio-control agent was evaluated in vitro against Roselle pathogens i.e. Phoma exigua, Fusarium nygamai and Rhizoctonia solani[1] using the dual culture technique. Volatile and non-volatile inhibitors of Trichoderma were also evaluated for this purpose. T. viride[2] was shown to have a marked inhibitory effect on the tested pathogens in vitro. Maximum inhibition occurred against P. exigua, with 71.76% reduction in mycelial radial growth. The three pathogens, P. exigua, F. nygamai and R. solani were also found to be susceptible to the volatile inhibitors produced by T. viride, giving rise to growth inhibition of about 68% in each case. When T. viride non-volatile metabolites were tested against the pathogens, maximum inhibition occurred against R. solani (73.95% mycelial growth inhibition), followed by P. exigua (37.17% inhibition). The inhibitory effect of the non-volatile metabolites on F. nygamai was, however, minimal. Copyright © 2012 Elsevier Ltd. All rights reserved.

Bacterial community analysis of swine manure treated with autothermal thermophilic aerobic digestion.

PubMed

Han, Il; Congeevaram, Shankar; Ki, Dong-Won; Oh, Byoung-Taek; Park, Joonhong

2011-02-01

Due to the environmental problems associated with disposal of livestock sludge, many stabilization studies emphasizing on the sludge volume reduction were performed. However, little is known about the microbial risk present in sludge and its stabilized products. This study microbiologically explored the effects of anaerobic lagoon fermentation (ALF) and autothermal thermophilic aerobic digestion (ATAD) on pathogen-related risk of raw swine manure by using culture-independent 16S rDNA cloning and sequencing methods. In raw swine manure, clones closely related to pathogens such as Dialister pneumosintes, Erysipelothrix rhusiopathiae, Succinivibrioan dextrinosolvens, and Schineria sp. were detected. Meanwhile, in the mesophilic ALF-treated swine manure, bacterial community clones closely related to pathogens such as Schineria sp. and Succinivibrio dextrinosolvens were still detected. Interestingly, the ATAD treatment resulted in no detection of clones closely related to pathogens in the stabilized thermophilic bacterial community, with the predominance of novel Clostridia class populations. These findings support the superiority of ATAD in selectively reducing potential human and animal pathogens compared to ALF, which is a typical manure stabilization method used in livestock farms.

Resistance in persisting bat populations after white-nose syndrome invasion.

PubMed

Langwig, Kate E; Hoyt, Joseph R; Parise, Katy L; Frick, Winifred F; Foster, Jeffrey T; Kilpatrick, A Marm

2017-01-19

Increases in anthropogenic movement have led to a rise in pathogen introductions and the emergence of infectious diseases in naive host communities worldwide. We combined empirical data and mathematical models to examine changes in disease dynamics in little brown bat (Myotis lucifugus) populations following the introduction of the emerging fungal pathogen Pseudogymnoascus destructans, which causes the disease white-nose syndrome. We found that infection intensity was much lower in persisting populations than in declining populations where the fungus has recently invaded. Fitted models indicate that this is most consistent with a reduction in the growth rate of the pathogen when fungal loads become high. The data are inconsistent with the evolution of tolerance or an overall reduced pathogen growth rate that might be caused by environmental factors. The existence of resistance in some persisting populations of little brown bats offers a glimmer of hope that a precipitously declining species will persist in the face of this deadly pathogen.This article is part of the themed issue 'Human influences on evolution, and the ecological and societal consequences'. © 2016 The Author(s).

Primary Hemostatic Capacity of Whole Blood: A Comprehensive Analysis of Pathogen Reduction and Refrigeration Effects Over Time

DTIC Science & Technology

2013-01-01

reduction and refrigeration is intriguing and suggests that PRT WB may complement the beneficial effects of the antifibrinolytic, tranexamic acid in...and ABBREVIATIONS: aPTT = activated partial thromboplastin time; ASPI = arachidonic acid agonist; COL = collagen; GPIba = glycoprotein Ib alpha; K...riboflavin and ultraviolet (UV) light to damage the nucleic acids of white blood cells (WBCs), parasites, bacteria, and viruses, is being consid- ered

Novel chitosan film embedded with liposome-encapsulated phage for biocontrol of Escherichia coli O157:H7 in beef.

PubMed

Cui, Haiying; Yuan, Lu; Lin, Lin

2017-12-01

In recent years, phages used for the reduction of pathogenic bacteria have fostered many attentions, but they are liable to lost bioactivity in food due to the presence of acidic compounds, enzymes and evaporite materials. To improve the stability of phages, a chitosan edible film containing liposome-encapsulated phage was engineered in the present study. The characteristics of liposome-encapsulated phage and the chitosan film containing liposome-encapsulated phage were investigated. The encapsulation efficiency of phages in liposome reached 57.66±0.12%. Besides, the desirable physical properties of chitosan film were obtained. The chitosan film embedded with liposome-encapsulated phage exhibited high antibacterial activity against Escherichia coli O157:H7, without the impact on the sensory properties of beef. Hence, chitosan film containing liposome-encapsulated phage could be a promising antibacterial packaging for beef preservation. Copyright © 2017 Elsevier Ltd. All rights reserved.

Inhibition of Candida albicans biofilm by pure selenium nanoparticles synthesized by pulsed laser ablation in liquids.

PubMed

Guisbiers, Grégory; Lara, Humberto H; Mendoza-Cruz, Ruben; Naranjo, Guillermo; Vincent, Brandy A; Peralta, Xomalin G; Nash, Kelly L

2017-04-01

Selenoproteins play an important role in the human body by accomplishing essential biological functions like oxido-reductions, antioxidant defense, thyroid hormone metabolism and immune response; therefore, the possibility to synthesize selenium nanoparticles free of any contaminants is exciting for future nano-medical applications. This paper reports the first synthesis of selenium nanoparticles by femtosecond pulsed laser ablation in de-ionized water. Those pure nanoparticles have been successfully used to inhibit the formation of Candida albicans biofilms. Advanced electron microscopy images showed that selenium nanoparticles easily adhere on the biofilm, then penetrate into the pathogen, and consequently damage the cell structure by substituting with sulfur. 50% inhibition of Candida albicans biofilm was obtained at only 25 ppm. Finally, the two physical parameters proved to affect strongly the viability of Candida albicans are the crystallinity and particle size. Copyright © 2016 Elsevier Inc. All rights reserved.

Disinfection potential of electrolyzed solutions containing sodium chloride at low concentrations.

PubMed

Morita, C; Sano, K; Morimatsu, S; Kiura, H; Goto, T; Kohno, T; Hong, W U; Miyoshi, H; Iwasawa, A; Nakamura, Y; Tagawa, M; Yokosuka, O; Saisho, H; Maeda, T; Katsuoka, Y

2000-03-01

Electrolyzed products of sodium chloride solution were examined for their disinfection potential against hepatitis B virus (HBV) and human immunodeficiency virus (HIV) in vitro. Electrolysis of 0.05% NaCl in tap water was carried out for 45 min at room temperature using a 3 A electric current in separate wells installed with positive and negative electrodes. The electrolyzed products were obtained from the positive well. The oxidation reduction potential (ORP), pH and free chlorine content of the product were 1053 mV, pH 2.34 and 4.20 ppm, respectively. The products modified the antigenicity of the surface protein of HBV as well as the infectivity of HIV in time- and concentration-dependent manner. Although the inactivating potential was decreased by the addition of contaminating protein, recycling of the product or continuous addition of fresh product may restore the complete disinfection against bloodborne pathogens.

The Effects of Signal Erosion and Core Genome Reduction on the Identification of Diagnostic Markers

PubMed Central

Sahl, Jason W.; Vazquez, Adam J.; Hall, Carina M.; Busch, Joseph D.; Tuanyok, Apichai; Mayo, Mark; Schupp, James M.; Lummis, Madeline; Pearson, Talima; Shippy, Kenzie; Allender, Christopher J.; Theobald, Vanessa; Hutcheson, Alex; Korlach, Jonas; LiPuma, John J.; Ladner, Jason; Lovett, Sean; Koroleva, Galina; Palacios, Gustavo; Limmathurotsakul, Direk; Wuthiekanun, Vanaporn; Wongsuwan, Gumphol; Currie, Bart J.

2016-01-01

ABSTRACT Whole-genome sequence (WGS) data are commonly used to design diagnostic targets for the identification of bacterial pathogens. To do this effectively, genomics databases must be comprehensive to identify the strict core genome that is specific to the target pathogen. As additional genomes are analyzed, the core genome size is reduced and there is erosion of the target-specific regions due to commonality with related species, potentially resulting in the identification of false positives and/or false negatives. PMID:27651357

Commensal or pathogen – a challenge to fulfil Koch’s Postulates

PubMed Central

Hess, M.

2017-01-01

ABSTRACT 1. Infectious diseases have a large impact on poultry health and economics. Elucidating the pathogenesis of a certain disease is crucial to implement control strategies. 2. Multiplication of a pathogen and its characterisation in vitro are basic requirements to perform experimental studies. However, passaging of the pathogen in vitro can influence the pathogenicity, a process targeted for live vaccine development, but limits the reproduction of clinical signs. 3. Numerous factors can influence the outcome of experimental infections with some importance on the pathogen, application route and host as exemplarily outlined for Histomonas meleagridis, Gallibacterium anatis and fowl aviadenoviruses (FAdVs). 4. In future, more comprehensive and detailed settings are needed to obtain as much information as possible from animal experiments. Processing of samples with modern diagnostic tools provides the option to closely monitor the host–pathogen interaction. PMID:27724044

Efficacy of vacuum steam pasteurization for inactivation of Salmonella PT 30, Escherichia coli O157:H7 and Enterococcus faecium on low moisture foods.

PubMed

Shah, Manoj K; Asa, Gladys; Sherwood, Julie; Graber, Kari; Bergholz, Teresa M

2017-03-06

Low moisture foods such as nuts, spices, and seeds have been implicated in several outbreaks due to Salmonella or E. coli O157:H7 contamination. Such foods may be consumed raw, and can be used as ingredients in other food products. While numerous thermal inactivation studies have been conducted for Salmonella on nuts, studies on other seeds and grains are minimal. Product water activity can influence the thermal resistance of pathogens, where thermal resistance increases as water activity decreases, leading to a requirement for higher temperatures and longer exposure times to achieve significant reduction of pathogen numbers. Vacuum steam pasteurization uses steam under vacuum, which can be operated at temperatures above and below 100°C. The objective of this study was to determine the efficacy of vacuum steam pasteurization for inactivation of pathogens on whole flaxseed, quinoa, sunflower kernels, milled flaxseed and whole black peppercorns. The use of E. faecium as a potential surrogate for Salmonella and E. coli O157:H7 in vacuum steam pasteurization was also evaluated. Pasteurization for 1min at 75°C yielded average log reductions of 5.48±1.22, 5.71±0.40 and 5.23±0.61 on flaxseed, 4.29±0.92, 5.89±0.26 and 2.39±0.83 on quinoa, and 4.01±0.74, 5.40±0.83 and 2.99±0.92 on sunflower kernels for Salmonella PT 30, E. coli O157:H7 and E. faecium, respectively. Similarly, on milled flaxseed and black peppercorns average log reductions of 3.02±0.79 and 6.10±0.64CFU/g were observed for Salmonella PT 30 after 1min of treatment at 75°C but, on average, >6.0 log reductions were observed after pasteurization at 85°C. Our data demonstrate that vacuum steam pasteurization can be effectively used to reduce pathogens on these low moisture foods at temperature as low as 75 and 85°C, and that E. faecium may be used as a potential surrogate for Salmonella PT 30 and E. coli O157:H7. Copyright © 2017 Elsevier B.V. All rights reserved.

Mastitis of periparturient Holstein cattle: a phenotypic and genetic study.

PubMed

Detilleux, J C; Kehrli, M E; Freeman, A E; Fox, L K; Kelley, D H

1995-10-01

Environmental and genetic factors affecting somatic cell scores, clinical mastitis, and IMI by minor and major pathogens were studied on 137 periparturient Holstein cows selected for milk production. Environmental effects were obtained by generalized least squares and logistic regression. Genetic parameters were from BLUP and threshold animal models. Lactation number affected the number of quarters with clinical mastitis and the number of quarters infected with minor pathogens. The DIM affected somatic cell score and number of quarters infected with major pathogens. Heritabilities for all mastitis indicators averaged 10%, but differences occurred among the indicators. Correlations between breeding values of the number of quarters infected with minor pathogens and the number infected with major pathogens were antagonistic and statistically significant.

Draft genome sequences of 50 MRSA ST5 isolates obtained from a U.S. hospital

USDA-ARS?s Scientific Manuscript database

Methicillin resistant Staphylococcus aureus (MRSA) can be a commensal or pathogen in humans. Pathogenicity and disease are related to the acquisition of mobile genetic elements encoding virulence and antimicrobial resistance genes. Here, we report draft genome sequences for 50 clinical MRSA isolates...

Green Fluorescent Protein (GFP) as a reporter gene for the plant pathogenic oomycete Phytophthora ramorum

Treesearch

Marko Riedel; Gautier Calmin; Lassaad Belbahri; Francois Lefort; Monika Gotz; Stefan Wagner; Sabine Werres

2009-01-01

Transgenic Phytophthora ramorum strains that produce green fluorescent protein (GFP) constitutively were obtained after stable DNA integration using a polyethylene glycol and CaCl2-based transformation protocol. Green fluorescent protein production was studied in developing colonies and in different propagules of the pathogen...

Molecular characterization and pathogenicity of fungal isolates for use against the small hive beetle (Aethina tumida)

USDA-ARS?s Scientific Manuscript database

The analysis of DNA sequences from fungal pathogens obtained from cadavers of the small hive beetle (SHB) collected from several apiaries in Florida revealed a mixture of saprobes and two potential primary entomopathogens, Metarhizium anisopliae and Beauveria bassiana. Spray tower bioassays indicate...

Bactericidal efficacy of atmospheric pressure non-thermal plasma (APNTP) against the ESKAPE pathogens.

PubMed

Flynn, Padrig B; Higginbotham, Sarah; Alshraiedeh, Nid'a H; Gorman, Sean P; Graham, William G; Gilmore, Brendan F

2015-07-01

The emergence of multidrug-resistant pathogens within the clinical environment is presenting a mounting problem in hospitals worldwide. The 'ESKAPE' pathogens (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa and Enterobacter spp.) have been highlighted as a group of causative organisms in a majority of nosocomial infections, presenting a serious health risk due to widespread antimicrobial resistance. The stagnating pipeline of new antibiotics requires alternative approaches to the control and treatment of nosocomial infections. Atmospheric pressure non-thermal plasma (APNTP) is attracting growing interest as an alternative infection control approach within the clinical setting. This study presents a comprehensive bactericidal assessment of an in-house-designed APNTP jet both against biofilms and planktonic bacteria of the ESKAPE pathogens. Standard plate counts and the XTT metabolic assay were used to evaluate the antibacterial effect of APNTP, with both methods demonstrating comparable eradication times. APNTP exhibited rapid antimicrobial activity against all of the ESKAPE pathogens in the planktonic mode of growth and provided efficient and complete eradication of ESKAPE pathogens in the biofilm mode of growth within 360s, with the exception of A. baumannii where a >4log reduction in biofilm viability was observed. This demonstrates its effectiveness as a bactericidal treatment against these pathogens and further highlights its potential application in the clinical environment for the control of highly antimicrobial-resistant pathogens. Copyright © 2015 Elsevier B.V. and the International Society of Chemotherapy. All rights reserved.

Lethality Prediction for Escherichia Coli O157:H7 and Uropathogenic E. coli in Ground Chicken Treated with High Pressure Processing and Trans-Cinnamaldehyde.

PubMed

Sheen, Shiowshuh; Huang, Chi-Yun; Ramos, Rommel; Chien, Shih-Yung; Scullen, O Joseph; Sommers, Christopher

2018-03-01

Pathogenic Escherichia coli, intestinal (O157:H7) as well as extraintestinal types (for example, Uropathogenic E. coli [UPEC]) are commonly found in many foods including raw chicken meat. The resistance of E. coli O157:H7 to UPEC in chicken meat under the stresses of high hydrostatic Pressure (HHP, also known as HPP-high pressure processing) and trans-cinnamaldehyde (an essential oil) was investigated and compared. UPEC was found slightly less resistant than O157:H7 in our test parameter ranges. With the addition of trans-cinnamaldehyde as an antimicrobial to meat, HPP lethality enhanced both O157:H7 and UPEC inactivation. To facilitate the predictive model development, a central composite design (CCD) was used to assess the 3-parameter effects, that is, pressure (300 to 400 MPa), trans-cinnamaldehyde dose (0.2 to 0.5%, w/w), and pressure-holding time (15 to 25 min), on the inactivation of E. coli O157:H7 and UPEC in ground chicken. Linear models were developed to estimate the lethality of E. coli O157:H7 (R 2 = 0.86) and UPEC (R 2 = 0.85), as well as dimensionless nonlinear models. All models were validated with data obtained from separated CCD combinations. Because linear models of O157:H7 and UPEC had similar R 2 and the significant lethality difference of CCD points was only 9 in 20; all data were combined to generate models to include both O157:H7 and UPEC. The results provide useful information/tool to predict how pathogenic E. coli may survive HPP in the presence of trans-cinnamaldehyde and to achieve a great than 5 log CFU/g reduction in chicken meat. The models may be used for process optimization, product development and to assist the microbial risk assessment. The study provided an effective means to reduce the high hydrostatic pressure level with incorporation of antimicrobial compound to achieve a 5-log reduction of pathogenic E. coli without damaging the raw meat quality. The developed models may be used to predict the high pressure processing lethality (and process optimization), product development (ingredient selection), and to assist the microbial risk assessment. © 2018 Institute of Food Technologists®.

Cold Plasma Inactivation of Bacterial Biofilms and Reduction of Quorum Sensing Regulated Virulence Factors

PubMed Central

Ziuzina, Dana; Boehm, Daniela; Patil, Sonal; Cullen, P. J.; Bourke, Paula

2015-01-01

The main objectives of this work were to investigate the effect of atmospheric cold plasma (ACP) against a range of microbial biofilms commonly implicated in foodborne and healthcare associated human infections and against P. aeruginosa quorum sensing (QS)-regulated virulence factors, such as pyocyanin, elastase (Las B) and biofilm formation capacity post-ACP treatment. The effect of processing factors, namely treatment time and mode of plasma exposure on antimicrobial activity of ACP were also examined. Antibiofilm activity was assessed for E. coli, L. monocytogenes and S. aureus in terms of reduction of culturability and retention of metabolic activity using colony count and XTT assays, respectively. All samples were treated ‘inpack’ using sealed polypropylene containers with a high voltage dielectric barrier discharge ACP generated at 80 kV for 0, 60, 120 and 300 s and a post treatment storage time of 24 h. According to colony counts, ACP treatment for 60 s reduced populations of E. coli to undetectable levels, whereas 300 s was necessary to significantly reduce populations of L. monocytogenes and S. aureus biofilms. The results obtained from XTT assay indicated possible induction of viable but non culturable state of bacteria. With respect to P. aeruginosa QS-related virulence factors, the production of pyocyanin was significantly inhibited after short treatment times, but reduction of elastase was notable only after 300 s and no reduction in actual biofilm formation was achieved post-ACP treatment. Importantly, reduction of virulence factors was associated with reduction of the cytotoxic effects of the bacterial supernatant on CHO-K1 cells, regardless of mode and duration of treatment. The results of this study point to ACP technology as an effective strategy for inactivation of established biofilms and may play an important role in attenuation of virulence of pathogenic bacteria. Further investigation is warranted to propose direct evidence for the inhibition of QS and mechanisms by which this may occur. PMID:26390435

Treatment of blood with a pathogen reduction technology using ultraviolet light and riboflavin inactivates Ebola virus in vitro.

PubMed

Cap, Andrew P; Pidcoke, Heather F; Keil, Shawn D; Staples, Hilary M; Anantpadma, Manu; Carrion, Ricardo; Davey, Robert A; Frazer-Abel, Ashley; Taylor, Audra L; Gonzales, Richard; Patterson, Jean L; Goodrich, Raymond P

2016-03-01

Transfusion of plasma from recovered patients after Ebolavirus (EBOV) infection, typically called "convalescent plasma," is an effective treatment for active disease available in endemic areas, but carries the risk of introducing other pathogens, including other strains of EBOV. A pathogen reduction technology using ultraviolet light and riboflavin (UV+RB) is effective against multiple enveloped, negative-sense, single-stranded RNA viruses that are similar in structure to EBOV. We hypothesized that UV+RB is effective against EBOV in blood products without activating complement or reducing protective immunoglobulin titers that are important for the treatment of Ebola virus disease (EVD). Four in vitro experiments were conducted to evaluate effects of UV+RB on green fluorescent protein EBOV (EBOV-GFP), wild-type EBOV in serum, and whole blood, respectively, and on immunoglobulins and complement in plasma. Initial titers for Experiments 1 to 3 were 4.21 log GFP units/mL, 4.96 log infectious units/mL, and 4.23 log plaque-forming units/mL. Conditions tested in the first three experiments included the following: 1-EBOV-GFP plus UV+RB; 2-EBOV-GFP plus RB only; 3-EBOV-GFP plus UV only; 4-EBOV-GFP without RB or UV; 5-virus-free control plus UV only; and 6-virus-free control without RB or UV. UV+RB reduced EBOV titers to nondetectable levels in both nonhuman primate serum (≥2.8- to 3.2-log reduction) and human whole blood (≥3.0-log reduction) without decreasing protective antibody titers in human plasma. Our in vitro results demonstrate that the UV+RB treatment efficiently reduces EBOV titers to below limits of detection in both serum and whole blood. In vivo testing to determine whether UV+RB can improve convalescent blood product safety is indicated. © 2016 AABB.

Treatment of blood with a pathogen reduction technology using UV light and riboflavin inactivates Ebola virus in vitro

PubMed Central

Cap, Andrew P.; Pidcoke, Heather F.; Keil, Shawn D.; Staples, Hilary M.; Anantpadma, Manu; Carrion, Ricardo; Davey, Robert A.; Frazer-Abel, Ashley; Taylor, Audra L.; Gonzales, Richard; Patterson, Jean L.; Goodrich, Raymond P.

2018-01-01

BACKGROUND Transfusion of plasma from recovered patients after Ebolavirus (EBOV) infection, typically called ‘convalescent plasma,’ is an effective treatment for active disease available in endemic areas, but carries the risk of introducing other pathogens, including other strains of EBOV. A pathogen reduction technology using ultraviolet light and riboflavin (UV + RB) is effective against multiple enveloped, negative-sense, single-stranded RNA viruses that are similar in structure to EBOV. We hypothesized that UV + RB is effective against EBOV in blood products without activating complement or reducing protective immunoglobulin titers that are important for the treatment of ebolavirus disease (EVD). STUDY DESIGN AND METHODS Four in vitro experiments were conducted to evaluate effects of UV + RB on green fluorescent protein EBOV (EBOV-GFP), wild-type EBOV in serum and whole blood, respectively, and on immunoglobulins and complement in plasma. Initial titers for Experiments 1–3 were: 4.21 log10 GFP units/mL, 4.96 log10 infectious units per mL, and 4.23 log10 plaque forming units per mL (PFU/mL). Conditions tested in the first three experiments included: 1. EBOV-GFP + UV + RB; 2. EBOV-GFP + RB only; 3 EBOV-GFP + UV only; 4. EBOV-GFP without RB or UV; 5. Virus-free control + UV only; and 6. Virus-free control without RB or UV. RESULTS UV + RB reduced EBOV titers to non-detectable levels in both non-human primate serum (≥ 2.8 to 3.2 log reduction) and human whole blood (≥ 3.0 log reduction) without decreasing protective antibody titers in human plasma. CONCLUSION Our in vitro results demonstrate that the UV + RB treatment efficiently reduces EBOV titers to below limits of detection in both serum and whole blood. In vivo testing to determine whether UV + RB can improve convalescent blood product safety is indicated. PMID:27001363

Pathogenicity of Leishmania donovani is associated with the high expression of a group low molecular weight proteins

PubMed Central

Mitra, Partha

2015-01-01

Background: With few exceptions, members of the Leishmania donovani complex such as L. donovani, L. infantum and L. chagashi are the etiological agents of visceral leishmaniasis or kala-azar. Promastigotes of Leishmania spp. lose their Pathogenicity; the ability to establish infection in a susceptible host, after prolonged culture. The molecular basis of this evolution of pathogenic to nonpathogenic culture has not been very well understood. It has been proposed that the loss of pathogenicity is associated with the gradual disappearance of selective parasite proteins. An alternative hypothesis is that during prolonged culture, the pathogenic clonal population of the parasite is deleted from the mixed population due to their selection pressure. This clonal deletion is proposed to be responsible for the emergence of the nonpathogenic population. Study Methodology and Results: We have a done a series of two-dimensional polyacrylamide gel electrophoresis followed by western blot experiments to study the antigenic profile of few L. donovani isolates of Indian origin. We observed a gradual and significant downregulation of expression of a group of low molecular weight proteins (LMW, molecular weight 20–30 kDa) which are associated with loss of pathogenicity. These proteins are recognized only by antiserum raised against the whole cell extract of one of the pathogenic Indian L. donovani isolates, Ag83, and remained undetected by antiserum raised against the nonpathogenic AG83 isolates. These LMW proteins were also present in the nonpathogenic extract in very low levels and remained undetected by the virulent serum, indicating a phenomenon of simultaneous downregulation of the expression and altered immunogenicity. LMW proteins were universally expressed in all early passage Indian isolate we tested and also detected in two clones obtained from pathogenic parasite culture. The antigenic patterns of none of the eight clones obtained from nonpathogenic culture were not exactly similar with the pathogenic clones. Conclusion: Therefore, our data strongly support the hypothesis that the loss of pathogenicity of L. donovani is associated with a change in antigenic profile, but not due the selective deletion of pathogenic clones. PMID:26629453

Pathogenicity of Leishmania donovani is associated with the high expression of a group low molecular weight proteins.

PubMed

Mitra, Partha

2015-01-01

With few exceptions, members of the Leishmania donovani complex such as L. donovani, L. infantum and L. chagashi are the etiological agents of visceral leishmaniasis or kala-azar. Promastigotes of Leishmania spp. lose their Pathogenicity; the ability to establish infection in a susceptible host, after prolonged culture. The molecular basis of this evolution of pathogenic to nonpathogenic culture has not been very well understood. It has been proposed that the loss of pathogenicity is associated with the gradual disappearance of selective parasite proteins. An alternative hypothesis is that during prolonged culture, the pathogenic clonal population of the parasite is deleted from the mixed population due to their selection pressure. This clonal deletion is proposed to be responsible for the emergence of the nonpathogenic population. We have a done a series of two-dimensional polyacrylamide gel electrophoresis followed by western blot experiments to study the antigenic profile of few L. donovani isolates of Indian origin. We observed a gradual and significant downregulation of expression of a group of low molecular weight proteins (LMW, molecular weight 20-30 kDa) which are associated with loss of pathogenicity. These proteins are recognized only by antiserum raised against the whole cell extract of one of the pathogenic Indian L. donovani isolates, Ag83, and remained undetected by antiserum raised against the nonpathogenic AG83 isolates. These LMW proteins were also present in the nonpathogenic extract in very low levels and remained undetected by the virulent serum, indicating a phenomenon of simultaneous downregulation of the expression and altered immunogenicity. LMW proteins were universally expressed in all early passage Indian isolate we tested and also detected in two clones obtained from pathogenic parasite culture. The antigenic patterns of none of the eight clones obtained from nonpathogenic culture were not exactly similar with the pathogenic clones. Therefore, our data strongly support the hypothesis that the loss of pathogenicity of L. donovani is associated with a change in antigenic profile, but not due the selective deletion of pathogenic clones.

Pathogen reduction in septic tank sludge through vermicomposting using Eisenia fetida.

PubMed

Rodríguez-Canché, L G; Cardoso Vigueros, L; Maldonado-Montiel, T; Martínez-Sanmiguel, M

2010-05-01

This study evaluated the potential of earthworms (Eisenia fetida) to remove pathogens from the sludge from septic tanks. Three earthworm population densities, equivalent to 1, 2, and 2.5kgm(-2), were tested for pathogen removal from sludge. The experimental phase lasted 60days, starting from the initial earthworm inoculation. After 60days, it was found that earthworms reduced concentrations of fecal coliforms, Salmonella spp., and helminth ova to permissible levels (<1000MPN/g, <3MPN/g, and <1viable ova/g on a dry weight basis, respectively) in accordance with Official Mexican Standard of environmental protection (NOM-004-SEMARNAT-2002) (SEMARNAT, 2002). Thus, sludge treatment with earthworms generated Class A biosolids, useful for forest, agricultural, and soil improvement. Copyright 2009 Elsevier Ltd. All rights reserved.

Specific detection of Xanthomonas axonopodis pv. dieffenbachiae in anthurium (Anthurium andreanum) tissues by nested PCR.

PubMed

Robène-Soustrade, Isabelle; Laurent, Philippe; Gagnevin, Lionel; Jouen, Emmanuel; Pruvost, Olivier

2006-02-01

Efficient control of Xanthomonas axonopodis pv. dieffenbachiae, the causal agent of anthurium bacterial blight, requires a sensitive and reliable diagnostic tool. A nested PCR test was developed from a sequence-characterized amplified region marker identified by randomly amplified polymorphic DNA PCR for the detection of X. axonopodis pv. dieffenbachiae. Serological and pathogenicity tests were performed concurrently with the nested PCR test with a large collection of X. axonopodis pv. dieffenbachiae strains that were isolated worldwide and are pathogenic to anthurium and/or other aroids. The internal primer pair directed amplification of the expected product (785 bp) for all 70 X. axonopodis pv. dieffenbachiae strains pathogenic to anthurium tested and for isolates originating from syngonium and not pathogenic to anthurium. This finding is consistent with previous studies which indicated that there is a high level of relatedness between strains from anthurium and strains from syngonium. Strains originating from the two host genera can be distinguished by restriction analysis of the amplification product. No amplification product was obtained with 98 strains of unrelated phytopathogenic bacteria or saprophytic bacteria from the anthurium phyllosphere, except for a weak signal obtained for one X. axonopodis pv. allii strain. Nevertheless, restriction enzyme analysis permitted the two pathovars to be distinguished. The detection threshold obtained with pure cultures or plant extracts (10(3) CFU ml(-1)) allowed detection of the pathogen from symptomless contaminated plants. This test could be a useful diagnostic tool for screening propagation stock plant material and for monitoring international movement of X. axonopodis pv. dieffenbachiae.

The Use of Filter-feeders to Manage Disease in a Changing World.

PubMed

Burge, Colleen A; Closek, Collin J; Friedman, Carolyn S; Groner, Maya L; Jenkins, Cody M; Shore-Maggio, Amanda; Welsh, Jennifer E

2016-10-01

Rapid environmental change is linked to increases in aquatic disease heightening the need to develop strategies to manage disease. Filter-feeding species are effective biofilters and can naturally mitigate disease risk to humans and wildlife. We review the role of filter-feeders, with an emphasis on bivalves, in altering disease outcomes via augmentation and reduction. Filtration can reduce transmission by removing pathogens from the water column via degradation and release of pathogens in pseudofeces. In other cases, filtration can increase pathogen transmission and disease risk. The effect of filtration on pathogen transmission depends on the selectivity of the filter-feeder, the degree of infectivity by the pathogen, the mechanism(s) of pathogen transmission and the ability of the pathogen to resist degradation. For example, some bacteria and viruses can resist degradation and accumulate within a filter-feeder leading to disease transmission to humans and other wildlife upon ingestion. Since bivalves can concentrate microorganisms, they are also useful as sentinels for the presence of pathogenic microorganisms. While somewhat less studied, other invertebrates, including ascidians and sponges may also provide ecosystem services by altering pathogen transmission. In all scenarios, climate change may affect the potential for filter-feeders to mitigate disease risk. We conclude that an assessment including empirical data and modeling of system-wide impacts should be conducted before selection of filter-feeders to mitigate disease. Such studies should consider physiology of the host and microbe and risk factors for negative impacts including augmentation of other pathogens. © The Author 2016. Published by Oxford University Press on behalf of the Society for Integrative and Comparative Biology. All rights reserved. For permissions please email: journals.permissions@oup.com.

Pathogen metadata platform: software for accessing and analyzing pathogen strain information.

PubMed

Chang, Wenling E; Peterson, Matthew W; Garay, Christopher D; Korves, Tonia

2016-09-15

Pathogen metadata includes information about where and when a pathogen was collected and the type of environment it came from. Along with genomic nucleotide sequence data, this metadata is growing rapidly and becoming a valuable resource not only for research but for biosurveillance and public health. However, current freely available tools for analyzing this data are geared towards bioinformaticians and/or do not provide summaries and visualizations needed to readily interpret results. We designed a platform to easily access and summarize data about pathogen samples. The software includes a PostgreSQL database that captures metadata useful for disease outbreak investigations, and scripts for downloading and parsing data from NCBI BioSample and BioProject into the database. The software provides a user interface to query metadata and obtain standardized results in an exportable, tab-delimited format. To visually summarize results, the user interface provides a 2D histogram for user-selected metadata types and mapping of geolocated entries. The software is built on the LabKey data platform, an open-source data management platform, which enables developers to add functionalities. We demonstrate the use of the software in querying for a pathogen serovar and for genome sequence identifiers. This software enables users to create a local database for pathogen metadata, populate it with data from NCBI, easily query the data, and obtain visual summaries. Some of the components, such as the database, are modular and can be incorporated into other data platforms. The source code is freely available for download at https://github.com/wchangmitre/bioattribution .

Introducing Pathogen Reduction Technology in Poland: A Cost-Utility Analysis

PubMed Central

Agapova, Maria; Lachert, Elzbieta; Brojer, Ewa; Letowska, Magdalena; Grabarczyk, Piotr; Custer, Brian

2015-01-01

Background Mirasol® pathogen reduction technology (PRT) uses UV light and riboflavin to chemically inactivate pathogens and white blood cells in blood components. In the EU, Mirasol PRT is CE-marked for both plasma and platelet treatment. In Poland, the decision to introduce PRT treatment of the national supply of fresh frozen plasma has spurred interest in evaluating the cost-effectiveness of this strategy. Methods A decision-analytic model evaluated the incremental costs and benefits of introducing PRT to the existing blood safety protocols in Poland. Results Addition of PRT treatment of plasma to current screening in Poland is estimated to cost 2.595 million PLN per quality-adjusted life year (QALY) (610,000 EUR/QALY); treating both plasma and platelet components in addition to current safety interventions had a lower cost of 1.480 million PLN/QALY (348,000 EUR/QALY). Conclusions The results suggest that in Poland the cost per QALY of PRT is high albeit lower than found in previous economic analyses of PRT and nucleic acid testing in North America. Treating both platelets and plasma components is more cost-effective than treating plasma alone. Wide confidence intervals indicate high uncertainty; to improve the precision of the health economic evaluation of PRT, additional hemovigilance data are needed. PMID:26195929

Local Context Finder (LCF) reveals multidimensional relationships among mRNA expression profiles of Arabidopsis responding to pathogen infection

PubMed Central

Katagiri, Fumiaki; Glazebrook, Jane

2003-01-01

A major task in computational analysis of mRNA expression profiles is definition of relationships among profiles on the basis of similarities among them. This is generally achieved by pattern recognition in the distribution of data points representing each profile in a high-dimensional space. Some drawbacks of commonly used pattern recognition algorithms stem from their use of a globally linear space and/or limited degrees of freedom. A pattern recognition method called Local Context Finder (LCF) is described here. LCF uses nonlinear dimensionality reduction for pattern recognition. Then it builds a network of profiles based on the nonlinear dimensionality reduction results. LCF was used to analyze mRNA expression profiles of the plant host Arabidopsis interacting with the bacterial pathogen Pseudomonas syringae. In one case, LCF revealed two dimensions essential to explain the effects of the NahG transgene and the ndr1 mutation on resistant and susceptible responses. In another case, plant mutants deficient in responses to pathogen infection were classified on the basis of LCF analysis of their profiles. The classification by LCF was consistent with the results of biological characterization of the mutants. Thus, LCF is a powerful method for extracting information from expression profile data. PMID:12960373

Predation Efficacy of Bdellovibrio bacteriovorus on Multidrug-Resistant Clinical Pathogens and Their Corresponding Biofilms.

PubMed

Sun, Yao; Ye, Jianzhong; Hou, Yuanbo; Chen, Huale; Cao, Jianming; Zhou, Tieli

2017-09-25

The aim of the present study was to evaluate the predation efficacy of Bdellovibrio bacteriovorus on multidrug-resistant (MDR) or extensive drug resistant (XDR) gram-negative pathogens and their corresponding biofilms. In this study, we examined the ability of B. bacteriovorus to prey on MDR and XDR gram-negative clinical bacteria, including Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Acinetobacter baumannii. Results showed that B. bacteriovorus was able to prey on all planktonic cultures, among which the most efficient predation was observed for drug-resistant E. coli, with a 3.11 log10 reduction in viability. Furthermore, B. bacteriovorus demonstrated promising efficacy in preventing biofilm formation and dispersing the established biofilm. Reductions in biofilm formation of E. coli, K. pneumoniae, P. aeruginosa, and A. baumannii co-cultured with B. bacteriovorus were 65.2%, 37.1%, 44.7%, and 36.8%, respectively. Meanwhile, the established biofilms of E. coli, K. pneumoniae, P. aeruginosa, and A. baumannii were significantly reduced by 83.4%, 81.8%, 83.1%, and 79.9%, respectively. A visual analysis supported by scanning electron microscopy demonstrated the role of B. bacteriovorus in removing the established biofilms. This study highlights the potential use of B. bacteriovorus as a biological control agent with the capability to prey on MDR/XDR gram-negative pathogens and eradicate biofilms.

Suppression of polygalacturonase gene expression in the phytopathogenic fungus Ophiostoma novo-ulmi by RNA interference.

PubMed

Carneiro, Joyce S; de la Bastide, Paul Y; Chabot, Meghan; Lerch, Lindsey; Hintz, William E

2010-05-01

The fungal pathogen, Ophiostomo novo-ulmi, has been responsible for the rapid decline of American elm (Ulmus americana) across North America and remains a serious threat to surviving elm populations. The production of pectinolytic polygalacturonase enzymes has been implicated as a virulence factor for many fungal pathogens, including O. novo-ulmi. Previous work has shown that the targeted disruption of the endopolygalacturonase gene locus epg1 of O. novo-ulmi reduced, but did not eliminate pectinase activity. In the present study, we evaluated the use of RNA interference (RNAi) as a method of suppressing expression of the epg1 locus in O. novo-ulmi and compared its efficiency to the gene disruption method. While there was a reduction in epg1-specific mRNA transcripts and in the amount of polygalacturonase enzyme secreted for both methods of gene regulation, neither method completely suppressed the expression of pectinase activity. There was, however, a significantly greater reduction in both transcript levels and secreted enzyme observed for some of the RNAi transformants. As the first demonstration of RNAi in O. novo-ulmi, this method of gene regulation shows promise in future studies of gene expression and pathogenicity. Copyright 2010 Elsevier Inc. All rights reserved.

Surfactin triggers biofilm formation of Bacillus subtilis in melon phylloplane and contributes to the biocontrol activity.

PubMed

Zeriouh, Houda; de Vicente, Antonio; Pérez-García, Alejandro; Romero, Diego

2014-07-01

The biocontrol activity of many Bacillus species has been traditionally related to the direct antagonism of pathogens. In previous works, we reported that B. subtilis strain UMAF6614 was an efficient biocontrol agent that produced bacillomycin, fengycin and surfactin lipopeptides. Bacillomycins and fengycins were shown to have antagonistic activity towards fungal and bacterial pathogens of cucurbits; however, the functionality of surfactin remained unclear. In this study, the role of surfactin in the biocontrol activity of this strain was investigated. We observed that a deficiency in surfactin production led to a partial reduction of disease suppression by this biocontrol agent, which coincided with a defect in biofilm formation and the colonization of the melon phylloplane. These effects were due to a dramatic reduction in the production of exopolysaccharide and the TasA protein, which are the two major components of the extracellular matrix. We propose that the biocontrol activity of this strain is the result of the coordinated action of the three families of lipopeptides. B. subtilis UMAF6614 produces surfactin to trigger biofilm formation on melon phylloplane, which ensures the long-term persistence and the adequate secretion of suppressive lipopeptides, bacillomycins and fengycins, which efficiently target pathogens. © 2013 Society for Applied Microbiology and John Wiley & Sons Ltd.

A field study using the fungicide benomyl to investigate the effect of mycorrhizal fungi on plant fitness.

PubMed

Carey, Peter D; Fitter, Alastair H; Watkinson, Andrew R

1992-07-01

The effect of vesicular-arbuscular mycorrhiza (VAM) on the fecundity ofVulpia ciliata ssp.ambigua was investigated at two field sites in eastern England by applying the fungicide benomyl to reduce VAM infection. The application of benomyl at the two sites produced very different results. At one site the application of the fungicide reduced the fecundity of plants whereas at the other fecundity was increased. At the first site the reduction in fecundity was linked to a significant reduction in VAM infection on the sprayed plants. The mechanism of the benefit associated with the VAM infection is however unclear: there was no treatment effect on morphology or on phosphorus inflow. At the second site, where fecundity was increased, there was only a negligible amount of VAM infection amongst the unsprayed plants and it is suggested that the increase in fecundity with the application of benomyl may have resulted from a reduction in infection by other, presumably pathogenic, fungi. The value of VAM fungi to the host plant may therefore not be restricted to physiological benefits. They may also provide protection to the plant by competing for space with other species of pathogenic fungi.

How Bacterial Pathogens Eat Host Lipids: Implications for the Development of Fatty Acid Synthesis Therapeutics*

PubMed Central

Yao, Jiangwei; Rock, Charles O.

2015-01-01

Bacterial type II fatty acid synthesis (FASII) is a target for the development of novel therapeutics. Bacteria incorporate extracellular fatty acids into membrane lipids, raising the question of whether pathogens use host fatty acids to bypass FASII and defeat FASII therapeutics. Some pathogens suppress FASII when exogenous fatty acids are present to bypass FASII therapeutics. FASII inhibition cannot be bypassed in many bacteria because essential fatty acids cannot be obtained from the host. FASII antibiotics may not be effective against all bacteria, but a broad spectrum of Gram-negative and -positive pathogens can be effectively treated with FASII inhibitors. PMID:25648887

Risk-Based Treatment Targets for Onsite Non-Potable Water Reuse

EPA Science Inventory

This presentation presents risk-based enteric pathogen log reduction targets for non-potable and potable uses of a variety of alternative source waters (i.e., municipal wastewater, locally-collected greywater, rainwater, and stormwater). A probabilistic, forward Quantitative Micr...

9 CFR 381.150 - Requirements for the production of fully cooked poultry products and partially cooked poultry...

Code of Federal Regulations, 2012 CFR

2012-01-01

... reduction of other pathogens and their toxins or toxic metabolites necessary to prevent adulteration, must... botulinum, and no more than a 1 log10 multiplication of Clostridium perfringens within the product. (b...

9 CFR 381.150 - Requirements for the production of fully cooked poultry products and partially cooked poultry...

Code of Federal Regulations, 2013 CFR

2013-01-01

... reduction of other pathogens and their toxins or toxic metabolites necessary to prevent adulteration, must... botulinum, and no more than a 1 log10 multiplication of Clostridium perfringens within the product. (b...

9 CFR 381.150 - Requirements for the production of fully cooked poultry products and partially cooked poultry...

Code of Federal Regulations, 2014 CFR

2014-01-01

... reduction of other pathogens and their toxins or toxic metabolites necessary to prevent adulteration, must... botulinum, and no more than a 1 log10 multiplication of Clostridium perfringens within the product. (b...

Entamoeba histolytica-secreted cysteine proteases induce IL-8 production in human mast cells via a PAR2-independent mechanism.

PubMed

Lee, Young Ah; Nam, Young Hee; Min, Arim; Kim, Kyeong Ah; Nozaki, Tomoyoshi; Saito-Nakano, Yumiko; Mirelman, David; Shin, Myeong Heon

2014-01-01

Entamoeba histolytica is an extracellular tissue parasite causing colitis and occasional liver abscess in humans. E. histolytica-derived secretory products (SPs) contain large amounts of cysteine proteases (CPs), one of the important amoebic virulence factors. Although tissue-residing mast cells play an important role in the mucosal inflammatory response to this pathogen, it is not known whether the SPs induce mast cell activation. In this study, when human mast cells (HMC-1 cells) were stimulated with SPs collected from pathogenic wild-type amoebae, interleukin IL-8 mRNA expression and production were significantly increased compared with cells incubated with medium alone. Inhibition of CP activity in the SPs with heat or the CP inhibitor E64 resulted in significant reduction of IL-8 production. Moreover, SPs obtained from inhibitors of cysteine protease (ICP)-overexpressing amoebae with low CP activity showed weaker stimulatory effects on IL-8 production than the wild-type control. Preincubation of HMC-1 cells with antibodies to human protease-activated receptor 2 (PAR2) did not affect the SP-induced IL-8 production. These results suggest that cysteine proteases in E. histolytica-derived secretory products stimulate mast cells to produce IL-8 via a PAR2-independent mechanism, which contributes to IL-8-mediated tissue inflammatory responses during the early phase of human amoebiasis. © Y.A. Lee et al., published by EDP Sciences, 2014.

Successful strategy for the selection of new strawberry-associated rhizobacteria antagonistic to Verticillium wilt.

PubMed

Berg, G; Kurze, S; Buchner, A; Wellington, E M; Smalla, K

2000-12-01

In order to isolate and characterize new strawberry-associated bacteria antagonistic to the soil-borne pathogenic fungus Verticillium dahliae Kleb., rhizobacterial populations from two different strawberry species, Greenish Strawberry (Fragaria viridis) and Garden Strawberry (F. x ananassa) obtained after plating onto King's B and glycerol-arginine agar, were screened for in vitro antagonism toward V. dahliae. The proportion of isolates with antifungal activity determined in in vitro assay against V. dahliae was higher for the Garden Strawberry than for the Greenish Strawberry. From 300 isolates, 20 isolates with strong antifungal activity were selected characterized by physiological profiling and molecular fingerprinting methods. Diversity among the isolates was characterized with molecular fingerprints using amplified ribosomal DNA restriction analysis (ARDRA) and the more discriminating BOX-PCR fingerprint method. The physiological profiles were well correlated with molecular fingerprinting pattern analysis. Significant reduction of Verticillium wilt by bacterial dipping bath treatment was shown in the greenhouse and in fields naturally infested by V. dahliae. The relative increase of yield ranged from 117% (Streptomyces albidoflavus S1) to 344% (Pseudomonas fluorescens P10) in greenhouse trials, and 113% (Streptomyces albidoflavus S1) to 247% (Pseudomonas fluorescens P6) in field trials. Evaluation resulted in the selection of three effective biocontrol agents (Pseudomonas fluorescens P6, P10, and Streptomyces diastatochromogenes S9) antagonistic to the Verticillium wilt pathogen.

Antifungal activity of food additives in vitro and as ingredients of hydroxypropyl methylcellulose-lipid edible coatings against Botrytis cinerea and Alternaria alternata on cherry tomato fruit.

PubMed

Fagundes, Cristiane; Pérez-Gago, María B; Monteiro, Alcilene R; Palou, Lluís

2013-09-16

The antifungal activity of food additives or 'generally recognized as safe' (GRAS) compounds was tested in vitro against Botrytis cinerea and Alternaria alternata. Radial mycelial growth of each pathogen was measured in PDA Petri dishes amended with food preservatives at 0.2, 1.0, or 2.0% (v/v) after 3, 5, and 7 days of incubation at 25 °C. Selected additives and concentrations were tested as antifungal ingredients of hydroxypropyl methylcellulose (HPMC)-lipid edible coatings. The curative activity of stable coatings was tested in in vivo experiments. Cherry tomatoes were artificially inoculated with the pathogens, coated by immersion about 24 h later, and incubated at 20 °C and 90% RH. Disease incidence and severity (lesion diameter) were determined after 6, 10, and 15 days of incubation and the 'area under the disease progress stairs' (AUDPS) was calculated. In general, HPMC-lipid antifungal coatings controlled black spot caused by A. alternata more effectively than gray mold caused by B. cinerea. Overall, the best results for reduction of gray mold on cherry tomato fruit were obtained with coatings containing 2.0% of potassium carbonate, ammonium phosphate, potassium bicarbonate, or ammonium carbonate, while 2.0% sodium methylparaben, sodium ethylparaben, and sodium propylparaben were the best ingredients for coatings against black rot. © 2013 Elsevier B.V. All rights reserved.

Effects of pathogen reduction systems on platelet microRNAs, mRNAs, activation, and function

PubMed Central

Osman, Abdimajid; Hitzler, Walter E.; Meyer, Claudius U.; Landry, Patricia; Corduan, Aurélie; Laffont, Benoit; Boilard, Eric; Hellstern, Peter; Vamvakas, Eleftherios C.

2015-01-01

Pathogen reduction (PR) systems for platelets, based on chemically induced cross-linking and inactivation of nucleic acids, potentially prevent transfusion transmission of infectious agents, but can increase clinically significant bleeding in some clinical studies. Here, we documented the effects of PR systems on microRNA and mRNA levels of platelets stored in the blood bank, and assessed their impact on platelet activation and function. Unlike platelets subjected to gamma irradiation or stored in additive solution, platelets treated with Intercept (amotosalen + ultraviolet-A [UVA] light) exhibited significantly reduced levels of 6 of the 11 microRNAs, and 2 of the 3 anti-apoptotic mRNAs (Bcl-xl and Clusterin) that we monitored, compared with platelets stored in plasma. Mirasol (riboflavin + UVB light) treatment of platelets did not produce these effects. PR neither affected platelet microRNA synthesis or function nor induced cross-linking of microRNA-sized endogenous platelet RNA species. However, the reduction in the platelet microRNA levels induced by Intercept correlated with the platelet activation (p < 0.05) and an impaired platelet aggregation response to ADP (p < 0.05). These results suggest that Intercept treatment may induce platelet activation, resulting in the release of microRNAs and mRNAs from platelets. The clinical implications of this reduction in platelet nucleic acids secondary to Intercept remain to be established. PMID:24749844

Extreme sensitivity to ultraviolet light in the fungal pathogen causing white-nose syndrome of bats.

PubMed

Palmer, Jonathan M; Drees, Kevin P; Foster, Jeffrey T; Lindner, Daniel L

2018-01-02

Bat white-nose syndrome (WNS), caused by the fungal pathogen Pseudogymnoascus destructans, has decimated North American hibernating bats since its emergence in 2006. Here, we utilize comparative genomics to examine the evolutionary history of this pathogen in comparison to six closely related nonpathogenic species. P. destructans displays a large reduction in carbohydrate-utilizing enzymes (CAZymes) and in the predicted secretome (~50%), and an increase in lineage-specific genes. The pathogen has lost a key enzyme, UVE1, in the alternate excision repair (AER) pathway, which is known to contribute to repair of DNA lesions induced by ultraviolet (UV) light. Consistent with a nonfunctional AER pathway, P. destructans is extremely sensitive to UV light, as well as the DNA alkylating agent methyl methanesulfonate (MMS). The differential susceptibility of P. destructans to UV light in comparison to other hibernacula-inhabiting fungi represents a potential "Achilles' heel" of P. destructans that might be exploited for treatment of bats with WNS.

Randomized controlled clinical trial evaluating multiplex polymerase chain reaction for pathogen identification and therapy adaptation in critical care patients with pulmonary or abdominal sepsis.

PubMed

Tafelski, Sascha; Nachtigall, Irit; Adam, Thomas; Bereswill, Stefan; Faust, Jana; Tamarkin, Andrey; Trefzer, Tanja; Deja, Maria; Idelevich, Evgeny A; Wernecke, Klaus-Dieter; Becker, Karsten; Spies, Claudia

2015-06-01

To determine whether a multiplex polymerase chain reaction (PCR)-based test could reduce the time required for initial pathogen identification in patients in an intensive care unit (ICU) setting. This double-blind, parallel-group randomized controlled trial** enrolled adults with suspected pulmonary or abdominal sepsis caused by an unknown pathogen. Both the intervention and control groups underwent the standard blood culture (BC) testing, but additional pathogen identification, based on the results of a LightCycler® SeptiFast PCR test, were provided in the intervention group. The study enrolled 37 patients in the control group and 41 in the intervention group. Baseline clinical and demographic characteristics were similar in both groups. The PCR-based test identified a pathogen in 10 out of 41 (24.4%) patients in the intervention group, with a mean duration from sampling to providing the information to the ICU of 15.9 h. In the control group, BC results were available after a significantly longer period (38.1 h). The LightCycler® SeptiFast PCR test demonstrated a significant reduction in the time required for initial pathogen identification, compared with standard BC. © The Author(s) 2015 Reprints and permissions: sagepub.co.uk/journalsPermissions.nav.

UV Light Inactivation of Human and Plant Pathogens in Unfiltered Surface Irrigation Water

PubMed Central

Jones, Lisa A.; Worobo, Randy W.

2014-01-01

Fruit and vegetable growers continually battle plant diseases and food safety concerns. Surface water is commonly used in the production of fruits and vegetables and can harbor both human- and plant-pathogenic microorganisms that can contaminate crops when used for irrigation or other agricultural purposes. Treatment methods for surface water are currently limited, and there is a need for suitable treatment options. A liquid-processing unit that uses UV light for the decontamination of turbid juices was analyzed for its efficacy in the treatment of surface waters contaminated with bacterial or oomycete pathogens, i.e., Escherichia coli, Salmonella enterica, Listeria monocytogenes, Clavibacter michiganensis subsp. michiganensis, Pseudomonas syringae pv. tomato, and Phytophthora capsici. Five-strain cocktails of each pathogen, containing approximately 108 or 109 CFU/liter for bacteria or 104 or 105 zoospores/liter for Ph. capsici, were inoculated into aliquots of two turbid surface water irrigation sources and processed with the UV unit. Pathogens were enumerated before and after treatment. In general, as the turbidity of the water source increased, the effectiveness of the UV treatment decreased, but in all cases, 99.9% or higher inactivation was achieved. Log reductions ranged from 10.0 to 6.1 and from 5.0 to 4.2 for bacterial pathogens and Ph. capsici, respectively. PMID:24242253

UV light inactivation of human and plant pathogens in unfiltered surface irrigation water.

PubMed

Jones, Lisa A; Worobo, Randy W; Smart, Christine D

2014-02-01

Fruit and vegetable growers continually battle plant diseases and food safety concerns. Surface water is commonly used in the production of fruits and vegetables and can harbor both human- and plant-pathogenic microorganisms that can contaminate crops when used for irrigation or other agricultural purposes. Treatment methods for surface water are currently limited, and there is a need for suitable treatment options. A liquid-processing unit that uses UV light for the decontamination of turbid juices was analyzed for its efficacy in the treatment of surface waters contaminated with bacterial or oomycete pathogens, i.e., Escherichia coli, Salmonella enterica, Listeria monocytogenes, Clavibacter michiganensis subsp. michiganensis, Pseudomonas syringae pv. tomato, and Phytophthora capsici. Five-strain cocktails of each pathogen, containing approximately 10(8) or 10(9) CFU/liter for bacteria or 10(4) or 10(5) zoospores/liter for Ph. capsici, were inoculated into aliquots of two turbid surface water irrigation sources and processed with the UV unit. Pathogens were enumerated before and after treatment. In general, as the turbidity of the water source increased, the effectiveness of the UV treatment decreased, but in all cases, 99.9% or higher inactivation was achieved. Log reductions ranged from 10.0 to 6.1 and from 5.0 to 4.2 for bacterial pathogens and Ph. capsici, respectively.

Isolation and identification of bacterial pathogen from mastitis milk in Central Java Indonesia

NASA Astrophysics Data System (ADS)

Harjanti, D. W.; Ciptaningtyas, R.; Wahyono, F.; Setiatin, ET

2018-01-01

Mastitis is a multi-etiologic disease of the mammary gland characterized mainly by reduction in milk production and milk quality due to intramammary infection by pathogenic bacteria. Nearly 83% of lactating dairy cows in Indonesia are infected with mastitis in various inflammation degrees. This study was conducted to isolate and identify the pathogen in milk collected from mastitis-infected dairy cows. The study was carried out in ten smallholder dairy farms in Central Java Indonesia based on animal examination, California mastitis test, isolation bacterial pathogens, Gram staining, Catalase and Coagulase test, and identification of bacteria species using Vitek. Bacteriological examination of milk samples revealed 15 isolates where Streptococcus was predominant species (73.3%) and the coagulase negative Staphylococcus species was identified at the least bacteria (26.7%). The Streptococcus bacteria found were Streptococcus uberis (2 isolates), Streptococcus sanguinis(6 isolates), Streptococcus dysgalactiaessp dysgalactiae(1 isolate) , Streptococcus mitis (1 isolate) and Streptococcus agalactiae (1 isolate). The Staphylococcus isolates comprising of Staphylococcus simulans (1 isolate) and Staphylococcus chromogens (3 isolates). Contamination of raw milkwith pathogenic bacteria can cause outbreaks of human disease (milk borne disease). Thus, proper milk processing method that couldinhibit the growth or kill these pathogenic bacteria is important to ensure the safety of milk and milk products.

Ectopic Expression of Xylella fastidiosa rpfF Conferring Production of Diffusible Signal Factor in Transgenic Tobacco and Citrus Alters Pathogen Behavior and Reduces Disease Severity.

PubMed

Caserta, R; Souza-Neto, R R; Takita, M A; Lindow, S E; De Souza, A A

2017-11-01

The pathogenicity of Xylella fastidiosa is associated with its ability to colonize the xylem of host plants. Expression of genes contributing to xylem colonization are suppressed, while those necessary for insect vector acquisition are increased with increasing concentrations of diffusible signal factor (DSF), whose production is dependent on RpfF. We previously demonstrated that transgenic citrus plants ectopically expressing rpfF from a citrus strain of X. fastidiosa subsp. pauca exhibited less susceptibility to Xanthomonas citri subsp. citri, another pathogen whose virulence is modulated by DSF accumulation. Here, we demonstrate that ectopic expression of rpfF in both transgenic tobacco and sweet orange also confers a reduction in disease severity incited by X. fastidiosa and reduces its colonization of those plants. Decreased disease severity in the transgenic plants was generally associated with increased expression of genes conferring adhesiveness to the pathogen and decreased expression of genes necessary for active motility, accounting for the reduced population sizes achieved in the plants, apparently by limiting pathogen dispersal through the plant. Plant-derived DSF signal molecules in a host plant can, therefore, be exploited to interfere with more than one pathogen whose virulence is controlled by DSF signaling.

Life history trade-offs and relaxed selection can decrease bacterial virulence in environmental reservoirs.

PubMed

Mikonranta, Lauri; Friman, Ville-Petri; Laakso, Jouni

2012-01-01

Pathogen virulence is usually thought to evolve in reciprocal selection with the host. While this might be true for obligate pathogens, the life histories of opportunistic pathogens typically alternate between within-host and outside-host environments during the infection-transmission cycle. As a result, opportunistic pathogens are likely to experience conflicting selection pressures across different environments, and this could affect their virulence through life-history trait correlations. We studied these correlations experimentally by exposing an opportunistic bacterial pathogen Serratia marcescens to its natural protist predator Tetrahymena thermophila for 13 weeks, after which we measured changes in bacterial traits related to both anti-predator defence and virulence. We found that anti-predator adaptation (producing predator-resistant biofilm) caused a correlative attenuation in virulence. Even though the direct mechanism was not found, reduction in virulence was most clearly connected to a predator-driven loss of a red bacterial pigment, prodigiosin. Moreover, life-history trait evolution was more divergent among replicate populations in the absence of predation, leading also to lowered virulence in some of the 'predator absent' selection lines. Together these findings suggest that the virulence of non-obligatory, opportunistic bacterial pathogens can decrease in environmental reservoirs through life history trade-offs, or random accumulation of mutations that impair virulence traits under relaxed selection.

Partial Tmem106b reduction does not correct abnormalities due to progranulin haploinsufficiency.

PubMed

Arrant, Andrew E; Nicholson, Alexandra M; Zhou, Xiaolai; Rademakers, Rosa; Roberson, Erik D

2018-06-22

Loss of function mutations in progranulin (GRN) are a major cause of frontotemporal dementia (FTD). Progranulin is a secreted glycoprotein that localizes to lysosomes and is critical for proper lysosomal function. Heterozygous GRN mutation carriers develop FTD with TDP-43 pathology and exhibit signs of lysosomal dysfunction in the brain, with increased levels of lysosomal proteins and lipofuscin accumulation. Homozygous GRN mutation carriers develop neuronal ceroid lipofuscinosis (NCL), an earlier-onset lysosomal storage disorder caused by severe lysosomal dysfunction. Multiple genome-wide association studies have shown that risk of FTD in GRN mutation carriers is modified by polymorphisms in TMEM106B, which encodes a lysosomal membrane protein. Risk alleles of TMEM106B may increase TMEM106B levels through a variety of mechanisms. Brains from FTD patients with GRN mutations exhibit increased TMEM106B expression, and protective TMEM106B polymorphisms are associated with decreased TMEM106B expression. Together, these data raise the possibility that reduction of TMEM106B levels may protect against the pathogenic effects of progranulin haploinsufficiency. We crossed Tmem106b +/- mice with Grn +/- mice, which model the progranulin haploinsufficiency of GRN mutation carriers and develop age-dependent social deficits and lysosomal abnormalities in the brain. We tested whether partial Tmem106b reduction could normalize the social deficits and lysosomal abnormalities of Grn +/- mice. Partial reduction of Tmem106b levels did not correct the social deficits of Grn +/- mice. Tmem106b reduction also failed to normalize most lysosomal abnormalities of Grn +/- mice, except for β-glucuronidase activity, which was suppressed by Tmem106b reduction and increased by progranulin insufficiency. These data do not support the hypothesis that Tmem106b reduction protects against the pathogenic effects of progranulin haploinsufficiency, but do show that Tmem106b reduction normalizes some lysosomal phenotypes in Grn +/- mice.

Inactivation of Heat Adapted and Chlorine Adapted Listeria Monocytogenes ATCC 7644 on Tomatoes Using Sodium Dodecyl Sulphate, Levulinic Acid and Sodium Hypochlorite Solution.

PubMed

Ijabadeniyi, Oluwatosin Ademola; Mnyandu, Elizabeth

2017-04-13

The effectiveness of sodium dodecyl sulphate (SDS), sodium hypochlorite solution and levulinic acid in reducing the survival of heat adapted and chlorine adapted Listeria monocytogenes ATCC 7644 was evaluated. The results against heat adapted L. monocytognes revealed that sodium hypochlorite solution was the least effective, achieving log reduction of 2.75, 2.94 and 3.97 log colony forming unit (CFU)/mL for 1, 3 and 5 minutes, respectively. SDS was able to achieve 8 log reduction for both heat adapted and chlorine adapted bacteria. When used against chlorine adapted L. monocytogenes sodium hypochlorite solution achieved log reduction of 2.76, 2.93 and 3.65 log CFU/mL for 1, 3 and 5 minutes, respectively. Using levulinic acid on heat adapted bacteria achieved log reduction of 3.07, 2.78 and 4.97 log CFU/mL for 1, 3, 5 minutes, respectively. On chlorine adapted bacteria levulinic acid achieved log reduction of 2.77, 3.07 and 5.21 log CFU/mL for 1, 3 and 5 minutes, respectively. Using a mixture of 0.05% SDS and 0.5% levulinic acid on heat adapted bacteria achieved log reduction of 3.13, 3.32 and 4.79 log CFU/mL for 1, 3 and 5 minutes while on chlorine adapted bacteria it achieved 3.20, 3.33 and 5.66 log CFU/mL, respectively. Increasing contact time also increased log reduction for both test pathogens. A storage period of up to 72 hours resulted in progressive log reduction for both test pathogens. Results also revealed that there was a significant difference (P≤0.05) among contact times, storage times and sanitizers. Findings from this study can be used to select suitable sanitizers and contact times for heat and chlorine adapted L. monocytogenes in the fresh produce industry.

[In vitro antibacterial activity of Curcuma longa (Zingiberaceae) against nosocomial bacteria in Montería, Colombia].

PubMed

Méndez Álvarez, Nelson; Angulo Ortíz, Alberto; Contreras Martínez, Orfa

2016-09-01

Bacterial resistance is a growing health problem worldwide that has serious economic and social impacts, compromising public health, and the therapeutic action of current antibiotics. Therefore, the search for new compounds with antimicrobial properties is relevant in modern studies, particularly against bacteria of clinical interest. In the present study, in vitro antibacterial activity of the ethanol extract and essential oil of Curcuma longa (Zingiberaceae) was evaluated against nosocomial bacteria, using the microdilution method. Escherichia coli strains, Pseudomonas aeruginosa, Klebsiella pneumoniae, Proteus sp. were used, Salmonella sp. and Bacillus sp., isolated from nosocomial infections in a hospital in the city of Monteria and reference strains of S. aureus ATCC 43300, S. aureus ATCC 29213, S. aureus ATCC 25923, P. aeruginosa ATCC 27853, E. coli ATCC 25922 and K. pneumonia ATCC 700603. The ethanol extract antibacterial profile was more efficient at higher concentrations (1 000 ppm), obtaining significant percentages of reduction of more than 50 % against K. pneumoniae ATCC 700603 and a clinical isolate of E. coli; while compared to Bacillus clinical isolate, was more active than the essential oil. For the rest of microorganisms, the reduction percentages obtained at a concentration of 1 000 ppm varied between 17 and 42 % with ethanolic extract, and 8 to 43 % with essential oil. At concentrations of 100 and 500 ppm antibacterial activity of the extracts was lower. The results indicated that the ethanolic extract and essential oil of C. longa rhizomes have active compounds with antibacterial properties that could be used in future research as a therapeutic alternative for the treatment of infections caused by nosocomial pathogens.

Effects of lactic acid and commercial chilling processes on survival of Salmonella, Yersinia enterocolitica, and Campylobacter coli in pork variety meats.

PubMed

King, Amanda M; Miller, Rhonda K; Castillo, Alejandro; Griffin, Davey B; Hardin, Margaret D

2012-09-01

Current industry chilling practices with and without the application of 2% L-lactic acid were compared for their effectiveness at reducing levels of Salmonella, Yersinia enterocolitica, and Campylobacter coli on pork variety meats. Pork variety meats (livers, intestines, hearts, and stomachs) were inoculated individually with one of the three pathogens and subjected to five different treatment combinations that included one or more of the following: water wash (25°C), lactic acid spray (2%, 40 to 50°C), chilling (4°C), and freezing (-15°C). Samples were analyzed before treatment, after each treatment step, and after 2, 4, and 6 months of frozen storage. Results showed that when a lactic acid spray was used in combination with water spray, immediate reductions were approximately 0.5 log CFU per sample of Salmonella, 0.8 log CFU per sample of Y. enterocolitica, and 1.1 log CFU per sample of C. coli. Chilling, both alone and in combination with spray treatments, had little effect on pathogens, while freezing resulted in additional 0.5-log CFU per sample reductions in levels of Salmonella and Y. enterocolitica, and an additional 1.0-log CFU per sample reduction in levels of C. coli. While reductions of at least 1 log CFU per sample were observed on variety meats treated with only a water wash and subsequently frozen, samples treated with lactic acid had greater additional reductions than those treated with only a water spray throughout frozen storage. The results of this study suggest that the use of lactic acid as a decontamination intervention, when used in combination with good manufacturing practices during processing, causes significant reductions in levels of Salmonella, Y. enterocolitica, and C. coli on pork variety meats.

Identification of the inactivating factors and mechanisms exerted on MS2 coliphage in concentrated synthetic urine.

PubMed

Oishi, Wakana; Sano, Daisuke; Decrey, Loic; Kadoya, Syunsuke; Kohn, Tamar; Funamizu, Naoyuki

2017-11-15

Volume reduction (condensation) is a key for the practical usage of human urine as a fertilizer because it enables the saving of storage space and the reduction of transportation cost. However, concentrated urine may carry infectious disease risks resulting from human pathogens frequently present in excreta, though the survival of pathogens in concentrated urine is not well understood. In this study, the inactivation of MS2 coliphage, a surrogate for single-stranded RNA human enteric viruses, in concentrated synthetic urine was investigated. The infectious titer reduction of MS2 coliphage in synthetic urine samples was measured by plaque assay, and the reduction of genome copy number was monitored by reverse transcription-quantitative PCR (RTqPCR). Among chemical-physical conditions such as pH and osmotic pressure, uncharged ammonia was shown to be the predominant factor responsible for MS2 inactivation, independently of urine concentration level. The reduction rate of the viral genome number varied among genome regions, but the comprehensive reduction rate of six genome regions was well correlated with that of the infectious titer of MS2 coliphage. This indicates that genome degradation is the main mechanism driving loss of infectivity, and that RT-qPCR targeting the six genome regions can be used as a culture-independent assay for monitoring infectivity loss of the coliphage in urine. MS2 inactivation rate constants were well predicted by a model using ion composition and speciation in synthetic urine samples, which suggests that MS2 infectivity loss can be estimated solely based on the solution composition, temperature and pH, without explicitly accounting for effects of osmotic pressure. Copyright © 2017 Elsevier B.V. All rights reserved.

Proteomic and oxi-proteomic response of apple to a compatible (p. expansum) and a non-host (p. digitatum) pathogen

USDA-ARS?s Scientific Manuscript database

Despite the current use of chemical fungicides, Penicillium expansum still is one of the most devastating pathogens of pome fruit. In particular, P. expansum enters tissues through wounds causing large economic losses worldwide. To obtain new rational and environmental friendly control alternative...

Sequestration and Scavenging of Iron in Infection

PubMed Central

Parrow, Nermi L.; Fleming, Robert E.

2013-01-01

The proliferative capability of many invasive pathogens is limited by the bioavailability of iron. Pathogens have thus developed strategies to obtain iron from their host organisms. In turn, host defense strategies have evolved to sequester iron from invasive pathogens. This review explores the mechanisms employed by bacterial pathogens to gain access to host iron sources, the role of iron in bacterial virulence, and iron-related genes required for the establishment or maintenance of infection. Host defenses to limit iron availability for bacterial growth during the acute-phase response and the consequences of iron overload conditions on susceptibility to bacterial infection are also examined. The evidence summarized herein demonstrates the importance of iron bioavailability in influencing the risk of infection and the ability of the host to clear the pathogen. PMID:23836822

Customizable PCR-microplate array for differential identification of multiple pathogens.

PubMed

Woubit, Abdela; Yehualaeshet, Teshome; Roberts, Sherrelle; Graham, Martha; Kim, Moonil; Samuel, Temesgen

2013-11-01

Customizable PCR-microplate arrays were developed for the rapid identification of Salmonella Typhimurium, Salmonella Saintpaul, Salmonella Typhi, Shigella dysenteriae, Escherichia coli O157:H7, Francisella tularensis subsp. tularensis, Francisella tularensis subsp. novicida, Vibrio cholerae, Vibrio parahaemolyticus, Yersinia pestis, and Yersinia pseudotuberculosis. Previously, we identified highly specific primers targeting each of these pathogens. Here, we report the development of customizable PCR-microplate arrays for simultaneous identification of the pathogens using the primers identified. A mixed aliquot of genomic DNA from 38 strains was used to validate three PCR-microplate array formats. Identical PCR conditions were used to run all the samples on the three formats. Specific amplifications were obtained on all three custom plates. In preliminary tests performed to evaluate the sensitivity of these assays in samples inoculated in the laboratory with Salmonella Typhimurium, amplifications were obtained from 1 g of beef hot dog inoculated at as low as 9 CFU/ml or from milk inoculated at as low as 78 CFU/ml. Such microplate arrays could be valuable tools for initial identification or secondary confirmation of contamination by these pathogens.

Fluorocycline TP-271 Is Potent against Complicated Community-Acquired Bacterial Pneumonia Pathogens

PubMed Central

Fyfe, Corey; O’Brien, William; Hackel, Meredith; Minyard, Mary Beth; Waites, Ken B.; Dubois, Jacques; Murphy, Timothy M.; Slee, Andrew M.; Weiss, William J.; Sutcliffe, Joyce A.

2017-01-01

ABSTRACT TP-271 is a novel, fully synthetic fluorocycline antibiotic in clinical development for the treatment of respiratory infections caused by susceptible and multidrug-resistant pathogens. TP-271 was active in MIC assays against key community respiratory Gram-positive and Gram-negative pathogens, including Streptococcus pneumoniae (MIC90 = 0.03 µg/ml), methicillin-sensitive Staphylococcus aureus (MSSA; MIC90 = 0.25 µg/ml), methicillin-resistant S. aureus (MRSA; MIC90 = 0.12 µg/ml), Streptococcus pyogenes (MIC90 = 0.03 µg/ml), Haemophilus influenzae (MIC90 = 0.12 µg/ml), and Moraxella catarrhalis (MIC90 ≤0.016 µg/ml). TP-271 showed activity (MIC90 = 0.12 µg/ml) against community-acquired MRSA expressing Panton-Valentine leukocidin (PVL). MIC90 values against Mycoplasma pneumoniae, Legionella pneumophila, and Chlamydia pneumoniae were 0.004, 1, and 4 µg/ml, respectively. TP-271 was efficacious in neutropenic and immunocompetent animal pneumonia models, generally showing, compared to the burden at the start of dosing, ~2 to 5 log10 CFU reductions against MRSA, S. pneumoniae, and H. influenzae infections when given intravenously (i.v.) and ~1 to 4 log10 CFU reductions when given orally (p.o.). TP-271 was potent against key community-acquired bacterial pneumonia (CABP) pathogens and was minimally affected, or unaffected, by tetracycline-specific resistance mechanisms and fluoroquinolone or macrolide drug resistance phenotypes. IMPORTANCE Rising resistance rates for macrolides, fluoroquinolones, and β-lactams in the most common pathogens associated with community-acquired bacterial pneumonia (CABP) are of concern, especially for cases of moderate to severe infections in vulnerable populations such as the very young and the elderly. New antibiotics that are active against multidrug-resistant Streptococcus pneumoniae and Staphylococcus aureus are needed for use in the empirical treatment of the most severe forms of this disease. TP-271 is a promising new fluorocycline antibiotic demonstrating in vitro potency and nonclinical efficacy by intravenous and oral administration against the major pathogens associated with moderate to severe CABP. PMID:28251179

Role of the lpxM lipid A biosynthesis pathway gene in pathogenicity of avian pathogenic Escherichia coli strain E058 in a chicken infection model.

PubMed

Xu, Huiqing; Ling, Jielu; Gao, Qingqing; He, Hongbo; Mu, Xiaohui; Yan, Zhen; Gao, Song; Liu, Xiufan

2013-10-25

Lipopolysaccharide (LPS) is a major surface component of avian pathogenic Escherichia coli (APEC), and is a possible virulence factor in avian infections caused by this organism. The contribution of the lpxM gene, which encodes a myristoyl transferase that catalyzes the final step in lipid A biosynthesis, to the pathogenicity of APEC has not previously been assessed. In this study, an isogenic lpxM mutant, E058ΔlpxM, was constructed in APEC O2 strain E058 and then characterized. Structural analysis of lipid A from the parental strain and derived mutant showed that E058ΔlpxM lacked one myristoyl (C14:0) on its lipid A molecules. No differences were observed between the mutant and wild-type in a series of tests including growth rate in different broths and ability to survive in specific-pathogen-free chicken serum. However, the mutant showed significantly reduced invasion and intracellular survival in the avian macrophage HD11 cell line (P<0.05). Nitric oxide production reduction (P<0.05) and cytokine gene expression downregulation (P<0.05 or P<0.01) also showed in HD11 treated with E058ΔlpxM-derived LPS compared with that in cells treated with E058-derived LPS at different times. Compared to the parental strain E058, E058ΔlpxM had a significant reduction in bacterial load in heart (P<0.01), liver (P<0.01), spleen (P<0.01), lung (P<0.05), and kidney (P<0.05) tissues. The histopathological lesions in visceral organs of birds challenged with the wild-type strain were more severe than in birds infected with the mutant. However, the E058ΔlpxM mutant showed a similar sensitivity pattern to the parental strain following exposure to several hydrophobic reagents. These results indicate that the lpxM gene is important for the pathogenicity and biological activity of APEC strain E058. Copyright © 2013 Elsevier B.V. All rights reserved.

Inactivation of Escherichia coli in milk and concentrated milk using pulsed-light treatment.

PubMed

Miller, B M; Sauer, A; Moraru, C I

2012-10-01

Pulsed light (PL) treatment has been viewed as an alternative to thermal treatments for the inactivation of pathogenic and spoilage microorganisms in recent years. The objectives of this study were to quantify the effectiveness of PL on inactivating Escherichia coli in cow milk and to evaluate the effect of total solids and fat content on inactivation. Samples of reconstituted milk with variable total solids levels (9.8, 25, and 45%) and commercial cow milk with different fat contents (skim milk, 2% fat, and whole milk) were inoculated with nonpathogenic E. coli ATCC 25922 at a concentration of 10(7)cfu/mL. One milliliter of the inoculated sample was placed in a thin layer in a glass chamber and exposed to PL doses of up to 14.9 J/cm(2), both in static mode and turbulent mode. Survivors were quantified using standard plate counting. All experiments were performed in triplicate. Pulsed light treatment of the concentrated milks of 25 and 45% solids content resulted in reductions of less than 1 log, even in turbulent mode, whereas for the milk with 9.8% solids content, reduction levels of 2.5 log cfu were obtained after treatment with 8.4 J/cm(2) in turbulent mode. In the skim milk, a 3.4 log cfu reduction at 14.9 J/cm(2) was obtained and a plateau of the inactivation curve typical of PL treatment was not achieved. Under the same conditions, both 2% and whole milk attained inactivation levels greater than 2.5 log cfu. These data indicate that PL is effective for the inactivation of E. coli in milk, but has limited effectiveness for microbial inactivation in concentrated milk, due to the absorption of light by the milk solids and shielding of the bacteria in the concentrated substrates. Milk fat also diminishes the effectiveness of PL to some extent, due to light-scattering effects. Copyright © 2012 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Toxicity and metabolism of nitroalkanes and substituted nitroalkanes

USDA-ARS?s Scientific Manuscript database

A series of low molecular weight nitro- containing compounds has recently been discovered to have a variety of biological activities including the reduction of anaerobic methane production in ruminant animals and activity against economically important human pathogens, including Salmonella sp. and s...

Methionine: a new biopesticide for use in mosquito management

USDA-ARS?s Scientific Manuscript database

Mosquito larvicides are an effective means of source reduction, controlling the population size so that the number of adult females that are present to bite and potentially spread pathogenic organisms is decreased. Currently utilized mosquito larvicides include insect growth regulators, organophosph...

Detection of bovine mastitis pathogens by loop-mediated isothermal amplification and an electrochemical DNA chip.

PubMed

Kawai, Kazuhiro; Inada, Mika; Ito, Keiko; Hashimoto, Koji; Nikaido, Masaru; Hata, Eiji; Katsuda, Ken; Kiku, Yoshio; Tagawa, Yuichi; Hayashi, Tomohito

2017-12-22

Bovine mastitis causes significant economic losses in the dairy industry. Effective prevention of bovine mastitis requires an understanding of the infection status of a pathogenic microorganism in a herd that has not yet shown clinical signs of mastitis and appropriate treatment specific for the pathogenic microorganism. However, bacterial identification by culture has drawbacks in that the sensitivity may be low and the procedure can be complex. In this study, we developed a genetic detection method to identify mastitis pathogens using a simple and highly sensitive electrochemical DNA chip which can specifically detect bacterial DNA in milk specimens. First, we selected microorganisms belonging to 12 families and/or genera associated with mastitis for which testing should be performed. Next, we optimized the conditions for amplifying microorganism DNA by loop-mediated isothermal amplification (LAMP) using 32 primers and the use of a DNA chip capable of measuring all pathogens simultaneously. Sample detection could be completed in just a few hours using this method. Comparison of the results obtained with our DNA chip method and those obtained by bacterial culture verified that when the culture method was set to 100%, the total positive concordance rate of the DNA chip was 85.0% and the total negative concordance rate was 86.9%. Furthermore, the proposed method allows both rapid and highly sensitive detection of mastitis pathogens. We believe that this method will contribute to the development of an effective mastitis control program.

Detection of bovine mastitis pathogens by loop-mediated isothermal amplification and an electrochemical DNA chip

PubMed Central

KAWAI, Kazuhiro; INADA, Mika; ITO, Keiko; HASHIMOTO, Koji; NIKAIDO, Masaru; HATA, Eiji; KATSUDA, Ken; KIKU, Yoshio; TAGAWA, Yuichi; HAYASHI, Tomohito

2017-01-01

Bovine mastitis causes significant economic losses in the dairy industry. Effective prevention of bovine mastitis requires an understanding of the infection status of a pathogenic microorganism in a herd that has not yet shown clinical signs of mastitis and appropriate treatment specific for the pathogenic microorganism. However, bacterial identification by culture has drawbacks in that the sensitivity may be low and the procedure can be complex. In this study, we developed a genetic detection method to identify mastitis pathogens using a simple and highly sensitive electrochemical DNA chip which can specifically detect bacterial DNA in milk specimens. First, we selected microorganisms belonging to 12 families and/or genera associated with mastitis for which testing should be performed. Next, we optimized the conditions for amplifying microorganism DNA by loop-mediated isothermal amplification (LAMP) using 32 primers and the use of a DNA chip capable of measuring all pathogens simultaneously. Sample detection could be completed in just a few hours using this method. Comparison of the results obtained with our DNA chip method and those obtained by bacterial culture verified that when the culture method was set to 100%, the total positive concordance rate of the DNA chip was 85.0% and the total negative concordance rate was 86.9%. Furthermore, the proposed method allows both rapid and highly sensitive detection of mastitis pathogens. We believe that this method will contribute to the development of an effective mastitis control program. PMID:29093278

Chemical composition, antibacterial and antifungal activities of essential oil from Cordia verbenacea DC leaves.

PubMed

Rodrigues, Fabiola F G; Oliveira, Liana G S; Rodrigues, Fábio F G; Saraiva, Manuele E; Almeida, Sheyla C X; Cabral, Mario E S; Campos, Adriana R; Costa, Jose Galberto M

2012-07-01

Cordia verbenacea is a Brazilian coastal shrub popularly known as "erva baleeira". The essential oil from fresh leaves was obtained by hydrodistillation and analyzed by CG/MS. The main components were identified as β-caryophyllene (25.4%), bicyclogermacrene (11.3%), δ-cadinene (9.%) and α-pinene (9.5%). In this study, the antimicrobial activity of Cordia verbenacea was evaluated. The minimal inhibitory concentration (MIC) of the essential oil was obtained using the broth microdilution assay (from 512 to 8 μg/ml). The results showed that the essential oil presented fungistatic activity against Candida albicans and Candida krusei and antibacterial activity against Gram-positive strains (Staphylococcus aureus and Bacillus cereus) and against multiresistant Gram-negative (Escherichia coli 27), in all tests the MIC was 64 μg/ml. When the essential oil was associated to aminoglycosides (subinhibitory concentrations, MIC/8), a synergic and antagonic activity was verified. The synergic effect was observed to the amikacin association (MIC reduction from 256 mlto 64 μg/ml) in all strains tested. The essential oil of Cordia verbenacea influences the activity of antibiotics and may be used as an adjuvant in antibiotic therapy against respiratory tract bacterial pathogens.

The first step toward genetic selection for host tolerance to infectious pathogens: obtaining the tolerance phenotype through group estimates

PubMed Central

Doeschl-Wilson, Andrea B.; Villanueva, Beatriz; Kyriazakis, Ilias

2012-01-01

Reliable phenotypes are paramount for meaningful quantification of genetic variation and for estimating individual breeding values on which genetic selection is based. In this paper, we assert that genetic improvement of host tolerance to disease, although desirable, may be first of all handicapped by the ability to obtain unbiased tolerance estimates at a phenotypic level. In contrast to resistance, which can be inferred by appropriate measures of within host pathogen burden, tolerance is more difficult to quantify as it refers to change in performance with respect to changes in pathogen burden. For this reason, tolerance phenotypes have only been specified at the level of a group of individuals, where such phenotypes can be estimated using regression analysis. However, few stsudies have raised the potential bias in these estimates resulting from confounding effects between resistance and tolerance. Using a simulation approach, we demonstrate (i) how these group tolerance estimates depend on within group variation and co-variation in resistance, tolerance, and vigor (performance in a pathogen free environment); and (ii) how tolerance estimates are affected by changes in pathogen virulence over the time course of infection and by the timing of measurements. We found that in order to obtain reliable group tolerance estimates, it is important to account for individual variation in vigor, if present, and that all individuals are at the same stage of infection when measurements are taken. The latter requirement makes estimation of tolerance based on cross-sectional field data challenging, as individuals become infected at different time points and the individual onset of infection is unknown. Repeated individual measurements of within host pathogen burden and performance would not only be valuable for inferring the infection status of individuals in field conditions, but would also provide tolerance estimates that capture the entire time course of infection. PMID:23412990

Advanced Analysis to Distinguish between Physical Decrease and Inactivation of Viable Phages in Aerosol by Quantitating Phage-Specific Particles.

PubMed

Shimasaki, Noriko; Nojima, Yasuhiro; Sakakibara, Masaya; Kikuno, Ritsuko; Iizuka, Chiori; Okaue, Akira; Okuda, Shunji; Shinohara, Katsuaki

2018-01-01

　Recent studies have investigated the efficacy of air-cleaning products against pathogens in the air. A standard method to evaluate the reduction in airborne viruses caused by an air cleaner has been established using a safe bacteriophage instead of pathogenic viruses; the reduction in airborne viruses is determined by counting the number of viable airborne phages by culture, after operating the air cleaner. The reduction in the number of viable airborne phages could be because of "physical decrease" or "inactivation". Therefore, to understand the mechanism of reduction correctly, an analysis is required to distinguish between physical decrease and inactivation. The purpose of this study was to design an analysis to distinguish between the physical decrease and inactivation of viable phi-X174 phages in aerosols. We established a suitable polymerase chain reaction (PCR) system by selecting an appropriate primer-probe set for PCR and validating the sensitivity, linearity, and specificity of the primer-probe set to robustly quantify phi-X174-specific airborne particles. Using this quantitative PCR system and culture assay, we performed a behavior analysis of the phage aerosol in a small chamber (1 m 3 ) at different levels of humidity, as humidity is known to affect the number of viable airborne phages. The results revealed that the reduction in the number of viable airborne phages was caused not only by physical decrease but also by inactivation under particular levels of humidity. Our study could provide an advanced analysis to differentiate between the physical decrease and inactivation of viable airborne phages.

Removal of phages and viral pathogens in a full-scale MBR: Implications for wastewater reuse and potable water.

PubMed

Purnell, Sarah; Ebdon, James; Buck, Austen; Tupper, Martyn; Taylor, Huw

2016-09-01

The aim of this study was to demonstrate how seasonal variability in the removal efficacy of enteric viral pathogens from an MBR-based water recycling system might affect risks to human health if the treated product were to be used for the augmentation of potable water supplies. Samples were taken over a twelve month period (March 2014-February 2015), from nine locations throughout a water recycling plant situated in East London and tested for faecal indicator bacteria (thermotolerant coliforms, intestinal enterococci n = 108), phages (somatic coliphage, F-specific RNA phage and Bacteroides phage (GB-124) n = 108), pathogenic viruses (adenovirus, hepatitis A, norovirus GI/GII n = 48) and a range of physico-chemical parameters (suspended solids, DO, BOD, COD). Thermotolerant coliforms and intestinal enterococci were removed effectively by the water recycling plant throughout the study period. Significant mean log reductions of 3.9-5.6 were also observed for all three phage groups monitored. Concentrations of bacteria and phages did not vary significantly according to season (P < 0.05; Kruskal-Wallis), though recorded levels of norovirus (GI) were significantly higher during autumn/winter months (P = 0.027; Kruskal-Wallis). Log reduction values for norovirus and adenovirus following MBR treatment were 2.3 and 4.4, respectively. However, both adenovirus and norovirus were detected at low levels (2000 and 3240 gene copies/L, respectively) post chlorination in single samples. Whilst phage concentrations did correlate with viral pathogens, the results of this study suggest that phages may not be suitable surrogates, as viral pathogen concentrations varied to a greater degree seasonally than did the phage indicators and were detected on a number of occasions on which phages were not detected (false negative sample results). Copyright © 2016 Elsevier Ltd. All rights reserved.

The gastrointestinal microbiome and its association with the control of pathogens in broiler chicken production: A review

PubMed Central

Clavijo, Viviana

2018-01-01

Abstract The microbiome of the broiler chicken gastrointestinal tract (GIT) has been extensively studied, and it has been amply demonstrated that it plays an important role in the health of the host, as it has a positive impact on the immune system, the physiology of the GIT, and productivity. Also, the microbiota is involved in reducing and preventing colonization by enteric pathogens through the process of competitive exclusion and the production of bacteriostatic and bactericidal substances. The taxonomic composition of the microbiota is affected by different factors, such as the organ, the age of the animal, diet and the use of antimicrobials. Different kinds of additives that regulate the microbial community in feed include probiotics (live microorganisms that when administered in adequate amounts confer a health benefit on the host), prebiotics (ingredients that stimulate increased beneficial microbial activity in the digestive system in order to improve the health of the host) and phytobiotics (primary or secondary components of plants that contain bioactive compounds that exert a positive effect on the growth and health of animals). Phages may potentially provide an integrated solution to modulate the intestinal microbiome of chicken intestines, as they reduce specific pathogenic microbial populations, permitting the proliferation of beneficial microbiota. Studies have shown that the use of cocktails of phages, especially in high concentrations and with short lapses of time between exposure to the bacteria and treatment with phages, optimize the reduction of Salmonella in chickens. Each of these technologies has demonstrable positive effects on the health of the host and the reduction of the pathogen load in controlled assays. This paper presents a comprehensive summary of the role of the microbiota in the broiler chicken gastrointestinal tract, and discusses the usefulness of different strategies for its modulation to control pathogens, with a particular emphasis on bacteriophages. PMID:29253263

Inactivation of Vibrio parahaemolyticus in hard clams (Mercanaria mercanaria) by high hydrostatic pressure (HHP) and the effect of HHP on the physical characteristics of hard clam meat.

PubMed

Mootian, Gabriel K; Flimlin, George E; Karwe, Mukund V; Schaffner, Donald W

2013-02-01

Shellfish may internalize dangerous pathogens during filter feeding. Traditional methods of depuration have been found ineffective against certain pathogens. The objective was to explore high hydrostatic pressure (HHP) as an alternative to the traditional depuration process. The effect of HHP on the survival of Vibrio parahaemolyticus in live clams (Mercanaria mercanaria) and the impact of HHP on physical characteristics of clam meat were investigated. Clams were inoculated with up to 7 log CFU/g of a cocktail of V. parahaemolyticus strains via filter feeding. Clams were processed at pressures ranging from 250 to 552 MPa for hold times ranging between 2 and 6 min. Processing conditions of 450 MPa for 4 min and 350 MPa for 6 min reduced the initial concentration of V. parahaemolyticus to a nondetectable level (<10(1) CFU/g), achieving >5 log reductions. The volume of clam meat (processed in shell) increased with negligible change in mass after exposure to pressure at 552 MPa for 3 min, while the drip loss was reduced. Clams processed at 552 MPa were softer compared to those processed at 276 MPa. However, all HHP processed clams were found to be harder compared to unprocessed. The lightness (L*) of the meat increased although the redness (a*) decreased with increasing pressure. Although high pressure-processed clams may pose a significantly lower risk from V. parahaemolyticus, the effect of the accompanied physical changes on the consumer's decision to purchase HHP clams remains to be determined. Shellfish may contain dangerous foodborne pathogens. Traditional methods of removing those pathogen have been found ineffective against certain pathogens. The objective of this research was to determine the effect of high hydrostatic pressure on V. parahaemolyticus in clams. Processing conditions of 450 MPa for 4 min and 350 MPa for 6 min reduced the initial concentration of V. parahaemolyticus to a nondetectable level, achieving >5 log reductions. © 2013 Institute of Food Technologists®

Antimicrobial effect of electrolyzed oxidizing water against Escherichia coli O157:H7 and Listeria monocytogenes on fresh strawberries (Fragaria x ananassa).

PubMed

Udompijitkul, P; Daeschel, M A; Zhao, Y

2007-11-01

Antibacterial activity of electrolyzed oxidizing (EO) water prepared from 0.05% or 0.10% (w/v) sodium chloride (NaCl) solutions against indigenous bacteria associated with fresh strawberries (Fragaria x ananassa) was evaluated. The efficacy of EO water and sodium hypochlorite (NaOCl) solution in eliminating and controlling the growth of Listeria monocytogenes and Escherichia coli O157:H7 inoculated onto strawberries stored at 4 +/- 1 degrees C up to 15 d was investigated at exposure time of 1, 5, or 10 min. Posttreatment neutralization of fruit surfaces was also determined. More than 2 log(10) CFU/g reductions of aerobic mesophiles were obtained in fruits washed for 10 or 15 min in EO water prepared from 0.10% (w/v) NaCl solution. Bactericidal activity of the disinfectants against L. monocytogenes and E. coli O157:H7 was not affected by posttreatment neutralization, and increasing exposure time did not significantly increase the antibacterial efficacy against both pathogens. While washing fruit surfaces with distilled water resulted in 1.90 and 1.27 log(10) CFU/mL of rinse fluid reduction of L. monocytogenes and E. coli O157:H7, respectively, >/= 2.60 log(10) CFU/mL of rinse fluid reduction of L. monocytogenes and up to 2.35 and 3.12 log(10) CFU/mL of rinse fluid reduction of E. coli O157:H7 were observed on fruit surfaces washed with EO water and NaOCl solution, respectively. Listeria monocytogenes and E. coli O157:H7 populations decreased over storage regardless of prior treatment. However, EO water and aqueous NaOCl did not show higher antimicrobial potential than water treatment during refrigeration storage.

Effect of oil gum massage therapy on common pathogenic oral microorganisms - A randomized controlled trial

PubMed Central

Singla, Nishu; Acharya, Shashidhar; Martena, Suganthi; Singla, Ritesh

2014-01-01

Objectives: (i) To assess reduction in Streptococcus mutans and Lactobacillus species count in saliva sample after ten minutes of oil gum massage therapy (massage of gingival tissues) per day for three weeks with sesame oil, olive oil, and coconut oil in three different groups of subjects. (ii) To compare the efficacy between three different oils and the “gold standard” chlorhexidine gel. (iii) To assess reduction in gingival scores and plaque scores of study subjects. Materials and Methods: Study design – Single center, parallel design, and triple blind randomized clinical study with four treatment groups. Participants: 32 of the 40 study subjects working as housekeeping personnel at Kasturba Hospital, Manipal; aged 18-55 years completed the three-week study period. Interventions: Subjects were randomly assigned to massage their gingiva everyday for three weeks with sesame oil, olive oil, coconut oil (tests), and Chlorhexidine gel (control). Oral health status and paraffin stimulated saliva samples were obtained at baseline and after three weeks of oil gum massage therapy. Outcome measures: Microbial culture, plaque index, and gingival index. Statistical analysis: Paired t test and Kruskal Wallis test. Results: There was a significant reduction in mean Streptococcus mutans count, Lactobacillus count, plaque scores, and gingival scores in all four groups after the study. However, there was no significant difference found in percentage reduction of these variables between the four groups. Conclusion: These oils can be used as valuable preventive agents in maintaining and improving oral health in low socioeconomic status population. However, it is recommended that further research should be conducted in other populations with a larger sample and longer duration of follow-up period. PMID:25210256

Physical Covering for Control of Escherichia coli O157:H7 and Salmonella spp. in Static and Windrow Composting Processes

PubMed Central

Yossa, Irene; Macarisin, Dumitru; Millner, Patricia

2015-01-01

This study investigated the effect of a 30-cm covering of finished compost (FC) on survival of Escherichia coli O157:H7 and Salmonella spp. in active static and windrow composting systems. Feedstocks inoculated with E. coli O157:H7 (7.41 log CFU/g) and Salmonella (6.46 log CFU/g) were placed in biosentry tubes (7.5-cm diameter, 30-cm height) at three locations: (i and ii) two opposing sides at the interface between the FC cover layer (where present) and the feedstock material (each positioned approximately 10 cm below the pile's surface) and (iii) an internal location (top) (approximately 30 cm below the surface). On specific sampling days, surviving populations of inoculated E. coli O157:H7 and Salmonella, generic E. coli, and coliforms in compost samples were determined. Salmonella spp. were reduced significantly within 24 h in windrow piles and were below the detection limit after 3 and 7 days at internal locations of windrow and static piles containing FC covering, respectively. Likewise, E. coli O157:H7 was undetectable after 1 day in windrow piles covered with finished compost. Use of FC as a covering layer significantly increased the number of days that temperatures in the windrows remained ≥55°C at all locations and in static piles at internal locations. These time-temperature exposures resulted in rapid reduction of inoculated pathogens, and the rate of bacterial reduction was rapid in windrow piles. The sample location significantly influenced the survival of these pathogens at internal locations compared to that at interface locations of piles. Finished compost covering of compost piles aids in the reduction of pathogens during the composting process. PMID:25576620

Genome dynamics and its impact on evolution of Escherichia coli.

PubMed

Dobrindt, Ulrich; Chowdary, M Geddam; Krumbholz, G; Hacker, J

2010-08-01

The Escherichia coli genome consists of a conserved part, the so-called core genome, which encodes essential cellular functions and of a flexible, strain-specific part. Genes that belong to the flexible genome code for factors involved in bacterial fitness and adaptation to different environments. Adaptation includes increase in fitness and colonization capacity. Pathogenic as well as non-pathogenic bacteria carry mobile and accessory genetic elements such as plasmids, bacteriophages, genomic islands and others, which code for functions required for proper adaptation. Escherichia coli is a very good example to study the interdependency of genome architecture and lifestyle of bacteria. Thus, these species include pathogenic variants as well as commensal bacteria adapted to different host organisms. In Escherichia coli, various genetic elements encode for pathogenicity factors as well as factors, which increase the fitness of non-pathogenic bacteria. The processes of genome dynamics, such as gene transfer, genome reduction, rearrangements as well as point mutations contribute to the adaptation of the bacteria into particular environments. Using Escherichia coli model organisms, such as uropathogenic strain 536 or commensal strain Nissle 1917, we studied mechanisms of genome dynamics and discuss these processes in the light of the evolution of microbes.

Evaluation of an automated ultraviolet-C light disinfection device and patient hand hygiene for reduction of pathogen transfer from interactive touchscreen computer kiosks.

PubMed

Alhmidi, Heba; Cadnum, Jennifer L; Piedrahita, Christina T; John, Amrita R; Donskey, Curtis J

2018-04-01

Touchscreens are a potential source of pathogen transmission. In our facility, patients and visitors rarely perform hand hygiene after using interactive touchscreen computer kiosks. An automated ultraviolet-C touchscreen disinfection device was effective in reducing bacteriophage MS2, bacteriophage ϕX174, methicillin-resistant Staphylococcus aureus, and Clostridium difficile spores inoculated onto a touchscreen. In simulations, an automated ultraviolet-C touchscreen disinfection device alone or in combination with hand hygiene reduced transfer of the viruses from contaminated touchscreens to fingertips. Published by Elsevier Inc.

Molecular assays reveal the presence and diversity of genes encoding pea footrot pathogenicity determinants in Nectria haematococca and in agricultural soils.

PubMed

Etebu, E; Osborn, A M

2009-05-01

The aim of this study was to develop molecular assays for investigating the presence and diversity of pathogenicity genes from the pea footrot pathogen Nectria haematococca (anamorph Fusarium solani f.sp. pisi) in soils. Polymerase chain reaction (PCR) assays were developed to amplify four N. haematococca pathogenicity genes (PDA, PEP1, PEP3 and PEP5) from isolates and soil-DNA from five agricultural fields with a prior footrot history. A collection of 15 fungi isolated on medium selective for Fusarium spp. exhibited variation in their virulence to peas as assessed via a disease index (DI: 0-5; no virulence to the highest virulence). PCR analyses showed that three isolates in which all four pathogenicity genes were detected resulted in the highest DI (>3.88). All four pathogenicity genes were detected in soil-DNA obtained from all five fields with a footrot disease history, but were not amplified from soils, which had no footrot history. Denaturing gradient gel electrophoresis and/or sequence analysis revealed diversity amongst the pathogenicity genes. The PCR assays developed herein enable the specific detection of pathogenic N. haematococca in soils without recourse to culture. Molecular assays that specifically target pathogenicity genes have the capacity to assess the presence of the footrot-causing pathogen in agricultural soils.

Role of anaerobic bacteria in biological soil disinfestation for elimination of soil-borne plant pathogens in agriculture.

PubMed

Ueki, Atsuko; Kaku, Nobuo; Ueki, Katsuji

2018-06-01

Biological soil disinfestation (BSD) or reductive soil disinfestation (RSD) is an environmental biotechnology to eliminate soil-borne plant pathogens based on functions of indigenous microbes. BSD treatments using different types of organic materials have been reported to effectively control a wide range of plant pathogens. Various studies have shown that development of reducing or anoxic conditions in soil is the most important aspect for effective BSD treatments. Substances such as organic acids, FeS, or phenolic compounds generated in the treated soil have been suggested to contribute to inactivation of pathogens. Additionally, anaerobic bacteria grown in the BSD-treated soil may produce and release enzymes with anti-pathogenic activities in soil. Clone library analyses as well as a next-generation sequence analysis based on 16S rRNA genes have revealed prosperity of obligate anaerobic bacteria from the class Clostridia in differently treated BSD soils. Two anaerobic bacterial strains isolated from BSD-treated soil samples and identified as Clostridium beijerinckii were found to decompose major cell wall polysaccharides of ascomycetous fungi, chitosan and β-1,3-glucan. C. beijerinckii is a species most frequently detected in the clone library analyses for various BSD-treated soils as a closely related species. The two anaerobic isolates severely degraded mycelial cells of the Fusarium pathogen of spinach wilt disease during anaerobic co-incubation of each isolate and the Fusarium pathogen. These reports suggest that antifungal enzymes produced by predominant anaerobic bacteria grown in the BSD-treated soil play important roles to control soil-borne fungal pathogens. Further studies using different bacterial isolates from BSD-treated soils are expected to know their anti-pathogenic abilities.

High‑throughput sequencing analyses of oral microbial diversity in healthy people and patients with dental caries and periodontal disease.

PubMed

Chen, Tingtao; Shi, Yan; Wang, Xiaolei; Wang, Xin; Meng, Fanjing; Yang, Shaoguo; Yang, Jian; Xin, Hongbo

2017-07-01

Recurrence of oral diseases caused by antibiotics has brought about an urgent requirement to explore the oral microbial diversity in the human oral cavity. In the present study, the high‑throughput sequencing method was adopted to compare the microbial diversity of healthy people and oral patients and sequence analysis was performed by UPARSE software package. The Venn results indicated that a mean of 315 operational taxonomic units (OTUs) was obtained, and 73, 64, 53, 19 and 18 common OTUs belonging to Firmicutes, Bacteroidetes, Proteobacteria, Actinobacteria and Fusobacteria, respectively, were identified in healthy people. Moreover, the reduction of Firmicutes and the increase of Proteobacteria in the children group, and the increase of Firmicutes and the reduction of Proteobacteria in the youth and adult groups, indicated that the age bracket and oral disease had largely influenced the tooth development and microbial development in the oral cavity. In addition, the traditional 'pathogenic bacteria' of Firmicutes, Proteobacteria and Bacteroidetes (accounted for >95% of the total sequencing number in each group) indicated that the 'harmful' bacteria may exert beneficial effects on oral health. Therefore, the data will provide certain clues for curing some oral diseases by the strategy of adjusting the disturbed microbial compositions in oral disease to healthy level.

Use of molecular markers to compare Fusarium verticillioides pathogenic strains isolated from plants and humans.

PubMed

Chang, S C; Macêdo, D P C; Souza-Motta, C M; Oliveira, N T

2013-08-12

Fusarium verticillioides is a pathogen of agriculturally important crops, especially maize. It is considered one of the most important pathogens responsible for fumonisin contamination of food products, which causes severe, chronic, and acute intoxication in humans and animals. Moreover, it is recognized as a cause of localized infections in immunocompetent patients and disseminated infections among severely immunosuppressed patients. Several molecular tools have been used to analyze the intraspecific variability of fungi. The objective of this study was to use molecular markers to compare pathogenic isolates of F. verticillioides and isolates of the same species obtained from clinical samples of patients with Fusarium mycoses. The molecular markers that we used were inter-simple sequence repeat markers (primers GTG5 and GACA4), intron splice site primer (primer EI1), random amplified polymorphic DNA marker (primer OPW-6), and restriction fragment length polymorphism-internal transcribed spacer (ITS) from rDNA. From the data obtained, clusters were generated based on the UPGMA clustering method. The amplification products obtained using primers ITS4 and ITS5 and loci ITS1-5.8-ITS2 of the rDNA yielded fragments of approximately 600 bp for all the isolates. Digestion of the ITS region fragment using restriction enzymes such as EcoRI, DraI, BshI, AluI, HaeIII, HinfI, MspI, and PstI did not permit differentiation among pathogenic and clinical isolates. The inter-simple sequence repeat, intron splice site primer, and random amplified polymorphic DNA markers presented high genetic homogeneity among clinical isolates in contrast to the high variability found among the phytopathogenic isolates of F. verticillioides.

The Reduction of Microbial and Chemical Contaminants with Selected POU/POE Systems

EPA Science Inventory

Centralized drinking water treatment and distribution alone may not always be the most practical or cost-effective option. Also, some consumers seeking a proactive measure to reduce exposure to pathogens and chemicals not currently monitored or regulated might consider employing...

Assessing Commercial and Alternative Poultry Processing Methods using Microbiome Analyses

USDA-ARS?s Scientific Manuscript database

Assessing poultry processing methods/strategies has historically used culture-based methods to assess bacterial changes or reductions, both in terms of general microbial communities (e.g. total aerobic bacteria) or zoonotic pathogens of interest (e.g. Salmonella, Campylobacter). The advent of next ...

Ability of select probiotics to reduce enteric Campylobacter colonization in broiler chickens

USDA-ARS?s Scientific Manuscript database

Campylobacter is the leading cause of foodborne enteritis worldwide and is primarily caused by consumption/mishandling of contaminated poultry. Probiotic use in poultry has been an effective strategy in reducing many enteric pathogens, but has not demonstrated consistent reduction against Campylobac...

PERKINSUS-"CIDAL" ACTIVITY OF OYSTER HEMOCYTES USING A TETRAZOLIUM DYE REDUCTION ASSAY: OPTIMIZATION AND APPLICATIONS

EPA Science Inventory

A bactericidal assay developed to assess the ability of oyster (Crassostrea virginica) hemocytes to kill the human pathogen Vibrio parahaemolyticus was optimized to estimate killing of the oyster parasite Perkinsus marinus. Assay variables, temperature, hemocyte:parasite ratio, i...

Biotechnological advances in the genetic improvement of Prunus domestica

USDA-ARS?s Scientific Manuscript database

Plum producers world-wide are facing multiple challenges including climate change, reductions in available labor, the need for reduced chemical inputs, the spread of native and exotic pests and pathogens, and consumer demands for improved fruit quality and health benefits. Meeting these challenges ...

Reduction of salmonella on valencia oranges by cold plasma treatment

USDA-ARS?s Scientific Manuscript database

Orange juice has been the source of recurrent food borne illness outbreaks, primarily associated with Salmonella. There is a need for antimicrobial interventions which can effectively eliminate pathogens from fruit surfaces and reduce the risk of cross-contamination during peeling and processing. To...

Selective decontamination of the digestive tract.

PubMed

Krueger, Wolfgang A; Unertl, Klaus E

2002-04-01

Ventilator-associated pneumonia usually originates from the patient's oropharyngeal microflora. In selective digestive decontamination, topical antibiotics are applied to the oropharynx and stomach for prevention of pneumonia and other infections, possibly reducing infection-related mortality. Selective digestive decontamination is also used for the prevention of gut-derived infections in acute necrotizing pancreatitis and liver transplantation. Despite numerous clinical trials, selective digestive decontamination remains controversial. Reduction of the incidence of pneumonia is accepted, but the extent of reduction is debated. Mortality was not reduced in most individual trials, but this finding was calculated in meta-analyses, especially for combined use of topical and systemic antibiotics in surgical ICU patients. Some investigators reported increased resistance and a shift to Gram-positive pathogens. Today, it appears that selective means not only selective suppression of pathogenic bacteria but also selection of appropriate groups of patients for underlying diseases and severity of illness, and selection of ICUs, where the endemic resistance patterns might allow the use of selective digestive decontamination at a relatively low risk for increased selection pressure.

Multiple-pass high-pressure homogenization of milk for the development of pasteurization-like processing conditions.

PubMed

Ruiz-Espinosa, H; Amador-Espejo, G G; Barcenas-Pozos, M E; Angulo-Guerrero, J O; Garcia, H S; Welti-Chanes, J

2013-02-01

Multiple-pass ultrahigh pressure homogenization (UHPH) was used for reducing microbial population of both indigenous spoilage microflora in whole raw milk and a baroresistant pathogen (Staphylococcus aureus) inoculated in whole sterile milk to define pasteurization-like processing conditions. Response surface methodology was followed and multiple response optimization of UHPH operating pressure (OP) (100, 175, 250 MPa) and number of passes (N) (1-5) was conducted through overlaid contour plot analysis. Increasing OP and N had a significant effect (P < 0·05) on microbial reduction of both spoilage microflora and Staph. aureus in milk. Optimized UHPH processes (five 202-MPa passes; four 232-MPa passes) defined a region where a 5-log(10) reduction of total bacterial count of milk and a baroresistant pathogen are attainable, as a requisite parameter for establishing an alternative method of pasteurization. Multiple-pass UHPH optimized conditions might help in producing safe milk without the detrimental effects associated with thermal pasteurization. © 2012 The Society for Applied Microbiology.

Ammonia sanitization of blackwater for safe use as fertilizer.

PubMed

Fidjeland, Jörgen; Svensson, Sven-Erik; Vinnerås, Björn

2015-01-01

Source-separated blackwater from low-flush toilets contains plant-available nutrients and can be used as a fertilizer. The aim of the study was to evaluate the impact on pathogen inactivation when treating blackwater with urea and/or lime. Blackwater was spiked with Salmonella typhimurium, Escherichia coli O157, Enterococcus faecalis, and Ascaris suum eggs, and treated with urea and/or lime in concentrations up to 0.1% w/w. The bottles were kept in a storage facility (manure slurry tank) for 102 days while monitoring the pathogen concentrations. The treatment time needed to meet the requirement for Salmonella and E. coli reduction could be reduced at least six-fold. The enterococci were more persistent, and only the highest treatment doses had a significantly higher inactivation than the controls. The Ascaris egg viability was only reduced by around 50%, so higher urea/lime doses and/or longer treatment times are required to fulfill the treatment requirements of 3 log10 reductions of parasite eggs.

Climate and land-use change impact on faecal indicator bacteria in a temperate maritime catchment (the River Conwy, Wales)

NASA Astrophysics Data System (ADS)

Bussi, Gianbattista; Whitehead, Paul G.; Thomas, Amy R. C.; Masante, Dario; Jones, Laurence; Jack Cosby, B.; Emmett, Bridget A.; Malham, Shelagh K.; Prudhomme, Christel; Prosser, Havard

2017-10-01

Water-borne pathogen contamination from untreated sewage effluent and runoff from farms is a serious threat to the use of river water for drinking and commercial purposes, such as downstream estuarine shellfish industries. In this study, the impact of climate change and land-use change on the presence of faecal indicator bacteria in freshwater was evaluated, through the use of a recently-developed catchment-scale pathogen model. The River Conwy in Wales has been used as a case-study, because of the large presence of livestock in the catchment and the importance of the shellfish harvesting activities in its estuary. The INCA-Pathogens catchment model has been calibrated through the use of a Monte-Carlo-based technique, based on faecal indicator bacteria measurements, and then driven by an ensemble of climate projections obtained from the HadRM3-PPE model (Future Flow Climate) plus four land-use scenarios (current land use, managed ecosystem, abandonment and agricultural intensification). The results show that climate change is not expected to have a very large impact on average river flow, although it might alter its seasonality. The abundance of faecal indicator bacteria is expected to decrease in response to climate change, especially during the summer months, due to reduced precipitation, causing reduced runoff, and increased temperature, which enhances the bacterial die-off processes. Land-use change can also have a potentially large impact on pathogens. The "managed ecosystems" scenario proposed in this study can cause a reduction of 15% in average water faecal indicator bacteria and up to 30% in the 90th percentile of water faecal indicator bacteria, mainly due to the conversion of pasture land into grassland and the expansion of forest land. This study provides an example of how to assess the impacts of human interventions on the landscape, and what may be the extent of their effects, for other catchments where the human use of the natural resources in the uplands can jeopardise the use of natural resources downstream.

Temperature effect on high salinity depuration of Vibrio vulnificus and V. parahaemolyticus from the Eastern oyster (Crassostrea virginica).

PubMed

Larsen, A M; Rikard, F S; Walton, W C; Arias, C R

2015-01-02

Vibrio vulnificus (Vv) and Vibrio parahaemolyticus (Vp) are opportunistic human pathogens naturally associated with the Eastern oyster Crassostrea virginica. The abundances of both pathogens in oysters are positively correlated with temperature, thus ingestion of raw oysters during the warm summer months is a risk factor for contracting illness from these bacteria. Current post-harvest processing (PHP) methods for elimination of these pathogens are expensive and kill the oyster, changing their organoleptic properties and making them less appealing to some consumers. High salinity has proven effective in reducing Vv numbers in the wild and our research aims at developing an indoor recirculating system to reduce pathogenic Vibrios while maintaining the taste and texture of live oysters. The goal of this study was to determine the influence of temperature on the efficacy of high salinity depuration. Vv was enumerated as most probable number (MPN) per gram of oyster tissue using the FDA-approved modified cellobiose polymyxin colistin (mCPC) protocol and with an alternative Vibrio specific media CHROMagar™ Vibrio (CaV). CaV was also used to quantify Vp. Oysters were held at 35 psu for 10 days at three temperatures: low (20°C), mid (22.5°C) and high (25°C). There was no difference in MPN/g of Vv between media; however more Vv isolates were obtained from mCPC than CaV. There was no significant effect of temperature on reduction of Vv or Vp throughout depuration but there was a tendency for low temperatures to be less effective than the higher ones. High salinity resulted in a significant decrease in Vv by day 3 and again by day 10, and a decrease in Vp by day 3. Oyster condition indices were maintained throughout depuration and mortality was low (4% across three trials). Overall these results support the use of mCPC for Vv enumeration and demonstrate the promise of high salinity depuration for PHP of the Eastern oyster. The trend for lower temperatures to be less effective is surprising and indicates a potential interaction between salinity and temperature that should be further investigated. Copyright © 2014. Published by Elsevier B.V.

Population subdivision of Fusarium graminearum from barley and wheat in the upper Midwestern United States at the turn of the century

USDA-ARS?s Scientific Manuscript database

Fusarium graminearum, the causal agent of Fusarium head blight (FHB) in wheat and barley, is one of the most economically destructive pathogens of these grains worldwide. Recent population genetic studies of the pathogen obtained from wheat in North America supported population subdivision in part c...

A dityrosine network mediated by dual oxidase and peroxidase influences the persistence of Lyme disease pathogens within the vector.

PubMed

Yang, Xiuli; Smith, Alexis A; Williams, Mark S; Pal, Utpal

2014-05-02

Ixodes scapularis ticks transmit a wide array of human and animal pathogens including Borrelia burgdorferi; however, how tick immune components influence the persistence of invading pathogens remains unknown. As originally demonstrated in Caenorhabditis elegans and later in Anopheles gambiae, we show here that an acellular gut barrier, resulting from the tyrosine cross-linking of the extracellular matrix, also exists in I. scapularis ticks. This dityrosine network (DTN) is dependent upon a dual oxidase (Duox), which is a member of the NADPH oxidase family. The Ixodes genome encodes for a single Duox and at least 16 potential peroxidase proteins, one of which, annotated as ISCW017368, together with Duox has been found to be indispensible for DTN formation. This barrier influences pathogen survival in the gut, as an impaired DTN in Doux knockdown or in specific peroxidase knockdown ticks, results in reduced levels of B. burgdorferi persistence within ticks. Absence of a complete DTN formation in knockdown ticks leads to the activation of specific tick innate immune pathway genes that potentially resulted in the reduction of spirochete levels. Together, these results highlighted the evolution of the DTN in a diverse set of arthropod vectors, including ticks, and its role in protecting invading pathogens like B. burgdorferi. Further understanding of the molecular basis of tick innate immune responses, vector-pathogen interaction, and their contributions in microbial persistence may help the development of new targets for disrupting the pathogen life cycle.

Targeted gene disruption of glycerol-3-phosphate dehydrogenase in Colletotrichum gloeosporioides reveals evidence that glycerol is a significant transferred nutrient from host plant to fungal pathogen.

PubMed

Wei, Yangdou; Shen, Wenyun; Dauk, Melanie; Wang, Feng; Selvaraj, Gopalan; Zou, Jitao

2004-01-02

Unidirectional transfer of nutrients from plant host to pathogen represents a most revealing aspect of the parasitic lifestyle of plant pathogens. Whereas much effort has been focused on sugars and amino acids, the identification of other significant metabolites is equally important for comprehensive characterization of metabolic interactions between plants and biotrophic fungal pathogens. Employing a strategy of targeted gene disruption, we generated a mutant strain (gpdhDelta) defective in glycerol-3-phosphate dehydrogenase in a hemibiotrophic plant pathogen, Colletotrichum gloeosporioides f.sp. malvae. The gpdhDelta strain had severe defects in carbon utilization as it could use neither glucose nor amino acids for sustained growth. Although the mutant mycelia were able to grow on potato dextrose agar medium, they displayed arrhythmicity in growth and failure to conidiate. The metabolic defect of gpdhDelta could be entirely ameliorated by glycerol in chemically defined minimal medium. Furthermore, glycerol was the one and only metabolite that could restore rhythmic growth and conidiation of gpdhDelta. Despite the profound defects in carbon source utilization, in planta the gpdhDelta strain exhibited normal pathogenicity, proceeded normally in its life cycle, and produced abundant conidia. Analysis of plant tissues at the peripheral zone of fungal infection sites revealed a time-dependent reduction in glycerol content. This study provides strong evidence for a role of glycerol as a significant transferred metabolite from plant to fungal pathogen.

Transmission of pathogens by Stomoxys flies (Diptera, Muscidae): a review

PubMed Central

Baldacchino, Frédéric; Muenworn, Vithee; Desquesnes, Marc; Desoli, Florian; Charoenviriyaphap, Theeraphap; Duvallet, Gérard

2013-01-01

Stomoxys flies are mechanical vectors of pathogens present in the blood and skin of their animal hosts, especially livestock, but occasionally humans. In livestock, their direct effects are disturbance, skin lesions, reduction of food intake, stress, blood loss, and a global immunosuppressive effect. They also induce the gathering of animals for mutual protection; meanwhile they favor development of pathogens in the hosts and their transmission. Their indirect effect is the mechanical transmission of pathogens. In case of interrupted feeding, Stomoxys can re-start their blood meal on another host. When injecting saliva prior to blood-sucking, they can inoculate some infected blood remaining on their mouthparts. Beside this immediate transmission, it was observed that Stomoxys may keep some blood in their crop, which offers a friendly environment for pathogens that could be regurgitated during the next blood meal; thus a delayed transmission by Stomoxys seems possible. Such a mechanism has a considerable epidemiological impact since it allows inter-herd transmission of pathogens. Equine infectious anemia, African swine fever, West Nile, and Rift Valley viruses are known to be transmitted by Stomoxys, while others are suspected. Rickettsia (Anaplasma, Coxiella), other bacteria and parasites (Trypanosoma spp., Besnoitia spp.) are also transmitted by Stomoxys. Finally, Stomoxys was also found to act as an intermediate host of the helminth Habronema microstoma and may be involved in the transmission of some Onchocerca and Dirofilaria species. Being cosmopolite, Stomoxys calcitrans might have a worldwide and greater impact than previously thought on animal and human pathogen transmission. PMID:23985165

Host-induced aneuploidy and phenotypic diversification in the Sudden Oak Death pathogen Phytophthora ramorum.

PubMed

Kasuga, Takao; Bui, Mai; Bernhardt, Elizabeth; Swiecki, Tedmund; Aram, Kamyar; Cano, Liliana M; Webber, Joan; Brasier, Clive; Press, Caroline; Grünwald, Niklaus J; Rizzo, David M; Garbelotto, Matteo

2016-05-20

Aneuploidy can result in significant phenotypic changes, which can sometimes be selectively advantageous. For example, aneuploidy confers resistance to antifungal drugs in human pathogenic fungi. Aneuploidy has also been observed in invasive fungal and oomycete plant pathogens in the field. Environments conducive to the generation of aneuploids, the underlying genetic mechanisms, and the contribution of aneuploidy to invasiveness are underexplored. We studied phenotypic diversification and associated genome changes in Phytophthora ramorum, a highly destructive oomycete pathogen with a wide host-range that causes Sudden Oak Death in western North America and Sudden Larch Death in the UK. Introduced populations of the pathogen are exclusively clonal. In California, oak (Quercus spp.) isolates obtained from trunk cankers frequently exhibit host-dependent, atypical phenotypes called non-wild type (nwt), apparently without any host-associated population differentiation. Based on a large survey of genotypes from different hosts, we previously hypothesized that the environment in oak cankers may be responsible for the observed phenotypic diversification in P. ramorum. We show that both normal wild type (wt) and nwt phenotypes were obtained when wt P. ramorum isolates from the foliar host California bay (Umbellularia californica) were re-isolated from cankers of artificially-inoculated canyon live oak (Q. chrysolepis). We also found comparable nwt phenotypes in P. ramorum isolates from a bark canker of Lawson cypress (Chamaecyparis lawsoniana) in the UK; previously nwt was not known to occur in this pathogen population. High-throughput sequencing-based analyses identified major genomic alterations including partial aneuploidy and copy-neutral loss of heterozygosity predominantly in nwt isolates. Chromosomal breakpoints were located at or near transposons. This work demonstrates that major genome alterations of a pathogen can be induced by its host species. This is an undocumented type of plant-microbe interaction, and its contribution to pathogen evolution is yet to be investigated, but one of the potential collateral effects of nwt phenotypes may be host survival.

Investigation of optimum ohmic heating conditions for inactivation of Escherichia coli O157:H7, Salmonella enterica serovar Typhimurium, and Listeria monocytogenes in apple juice.

PubMed

Park, Il-Kyu; Ha, Jae-Won; Kang, Dong-Hyun

2017-05-19

Control of foodborne pathogens is an important issue for the fruit juice industry and ohmic heating treatment has been considered as one of the promising antimicrobial interventions. However, to date, evaluation of the relationship between inactivation of foodborne pathogens and system performance efficiency based on differing soluble solids content of apple juice during ohmic heating treatment has not been well studied. This study aims to investigate effective voltage gradients of an ohmic heating system and corresponding sugar concentrations (°Brix) of apple juice for inactivating major foodborne pathogens (E. coli O157:H7, S. Typhimurium, and L. monocytogenes) while maintaining higher system performance efficiency. Voltage gradients of 30, 40, 50, and 60 V/cm were applied to 72, 48, 36, 24, and 18 °Brix apple juices. At all voltage levels, the lowest heating rate was observed in 72 °Brix apple juice and a similar pattern of temperature increase was shown in18-48 °Brix juice samples. System performance coefficients (SPC) under two treatment conditions (30 V/cm in 36 °Brix or 60 V/cm in 48 °Brix juice) were relatively greater than for other combinations. Meanwhile, 5-log reductions of the three foodborne pathogens were achieved after treatment for 60 s in 36 °Brix at 30 V/cm, but this same reduction was observed in 48 °Brix juice at 60 V/cm within 20 s without affecting product quality. With respect to both bactericidal efficiency and SPC values, 60 V/cm in 48 °Brix was the most effective ohmic heating treatment combination for decontaminating apple juice concentrates.

Practical Bench Comparison of BBL CHROMagar Orientation and Standard Two-Plate Media for Urine Cultures

PubMed Central

D'Souza, Holly A.; Campbell, Mary; Baron, Ellen Jo

2004-01-01

A total of 1,023 urine samples sent for routine culture were plated onto sheep blood and MacConkey agars and a BBL CHROMagar Orientation (CO; Becton Dickinson, Cockeysville, Md.) plate, and the results were compared. Of these, 250 urine samples (24%) grew >10,000 CFU of one or two putative pathogens/ml and 773 showed no growth (NG), mixed growth of <10,000 CFU/ml, or three or more strains (mixed). The CO and conventional medium results agreed completely for 595 cultures with NG or <10,000 CFU/ml. An additional 178 urine samples yielded clinically insignificant differences. Both medium sets essentially agreed on quantities and identification for 400 single-pathogen cultures and 9 mixed cultures. With the caveat that CO cannot differentiate Klebsiella, Enterobacter, and Serratia spp., enteric pathogens were identified only by morphology and color on CO. Direct visual differentiation of group B streptococci from lactobacilli is not possible, but lactobacillus cells always exhibited easily recognizable morphology on Gram stain. Of 108 paired organism susceptibility results encompassing 2,268 drug-pathogen combinations, there were 3% errors and only 1% very major errors. Use of CO allowed a >50% reduction in inoculation time and a >20% reduction in work-up time. For our laboratory, with 50% “no growth” and ca. 25% significant results (50% Escherichia coli), CO allowed time and workup cost savings for a majority of cultures. A cost analysis (time and supplies for our laboratory) showed that if CO is used alone, the break-even level for CO pricing is $1.78; if CO and blood agar are both used, the break-even pricing for CO is $1.53. PMID:14715732

General Suppression of Escherichia coli O157:H7 in Sand-Based Dairy Livestock Bedding▿ †

PubMed Central

Westphal, Andreas; Williams, Michele L.; Baysal-Gurel, Fulya; LeJeune, Jeffrey T.; McSpadden Gardener, Brian B.

2011-01-01

Sand bedding material is frequently used in dairy operations to reduce the occurrence of mastitis and enhance cow comfort. One objective of this work was to determine if sand-based bedding also supported the microbiologically based suppression of an introduced bacterial pathogen. Bedding samples were collected in summer, fall, and winter from various locations within a dairy operation and tested for their ability to suppress introduced populations of Escherichia coli O157:H7. All sources of bedding displayed a heat-sensitive suppressiveness to the pathogen. Differences in suppressiveness were also noted between different samples at room temperature. At just 1 day postinoculation (dpi), the recycled sand bedding catalyzed up to a 1,000-fold reduction in E. coli counts, typically 10-fold greater than the reduction achieved with other substrates, depending on the sampling date. All bedding substrates were able to reduce E. coli populations by over 10,000-fold within 7 to 15 dpi, regardless of sampling date. Terminal restriction fragment length polymorphism (T-RFLP) analysis was used to identify bacterial populations potentially associated with the noted suppression of E. coli O157:H7 in sand bedding. Eleven terminal restriction fragments (TRFs) were overrepresented in paired comparisons of suppressive and nonsuppressive specimens at multiple sampling points, indicating that they may represent environmentally stable populations of pathogen-suppressing bacteria. Cloning and sequencing of these TRFs indicated that they represent a diverse subset of bacteria, belonging to the Cytophaga-Flexibacter-Bacteroidetes, Gammaproteobacteria, and Firmicutes, only a few of which have previously been identified in livestock manure. Such data indicate that microbial suppression may be harnessed to develop new options for mitigating the risk and dispersal of zoonotic bacterial pathogens on dairy farms. PMID:21257815

A Presenilin-1 Mutation Identified in Familial Alzheimer Disease with Cotton Wool Plaques Causes a Nearly Complete Loss of γ-Secretase Activity*

PubMed Central

Heilig, Elizabeth A.; Xia, Weiming; Shen, Jie; Kelleher, Raymond J.

2010-01-01

Mutations in presenilin-1 and presenilin-2 (PS1 and PS2) are the most common cause of familial Alzheimer disease. PS1 and PS2 are the presumptive catalytic components of the multisubunit γ-secretase complex, which proteolyzes a number of type I transmembrane proteins, including the amyloid precursor protein (APP) and Notch. APP processing by γ-secretase produces β-amyloid peptides (Aβ40 and Aβ42) that accumulate in the Alzheimer disease brain. Here we identify a pathogenic L435F mutation in PS1 in two affected siblings with early-onset familial Alzheimer disease characterized by deposition of cerebral cotton wool plaques. The L435F mutation resides in a conserved C-terminal PAL sequence implicated in active site conformation and catalytic activity. The impact of PS1 mutations in and around the PAL motif on γ-secretase activity was assessed by expression of mutant PS1 in mouse embryo fibroblasts lacking endogenous PS1 and PS2. Surprisingly, the L435F mutation caused a nearly complete loss of γ-secretase activity, including >90% reductions in the generation of Aβ40, Aβ42, and the APP and Notch intracellular domains. Two nonpathogenic PS1 mutations, P433L and L435R, caused essentially complete loss of γ-secretase activity, whereas two previously identified pathogenic PS1 mutations, P436Q and P436S, caused partial loss of function with substantial reductions in production of Aβ40, Aβ42, and the APP and Notch intracellular domains. These results argue against overproduction of Aβ42 as an essential property of presenilin proteins bearing pathogenic mutations. Rather, our findings provide support for the hypothesis that pathogenic mutations cause a general loss of presenilin function. PMID:20460383

Different cross protection scopes of two avian influenza H5N1 vaccines against infection of layer chickens with a heterologous highly pathogenic virus.

PubMed

Poetri, Okti Nadia; Van Boven, Michiel; Koch, Guus; Stegeman, Arjan; Claassen, Ivo; Wayan Wisaksana, I; Bouma, Annemarie

2017-10-01

Avian influenza (AI) virus strains vary in antigenicity, and antigenic differences between circulating field virus and vaccine virus will affect the effectiveness of vaccination of poultry. Antigenic relatedness can be assessed by measuring serological cross-reactivity using haemagglutination inhibition (HI) tests. Our study aims to determine the relation between antigenic relatedness expressed by the Archetti-Horsfall ratio, and reduction of virus transmission of highly pathogenic H5N1 AI strains among vaccinated layers. Two vaccines were examined, derived from H5N1 AI virus strains A/Ck/WJava/Sukabumi/006/2008 and A/Ck/CJava/Karanganyar/051/2009. Transmission experiments were carried out in four vaccine and two control groups, with six sets of 16 specified pathogen free (SPF) layer chickens. Birds were vaccinated at 4weeks of age with one strain and challenge-infected with the homologous or heterologous strain at 8weeks of age. No transmission or virus shedding occurred in groups challenged with the homologous strain. In the group vaccinated with the Karanganyar strain, high cross-HI responses were observed, and no transmission of the Sukabumi strain occurred. However, in the group vaccinated with the Sukabumi strain, cross-HI titres were low, virus shedding was not reduced, and multiple transmissions to contact birds were observed. This study showed large differences in cross-protection of two vaccines based on two different highly pathogenic H5N1 virus strains. This implies that extrapolation of in vitro data to clinical protection and reduction of virus transmission might not be straightforward. Copyright © 2017 Elsevier Ltd. All rights reserved.

Identification of blood culture isolates directly from positive blood cultures by use of matrix-assisted laser desorption ionization-time of flight mass spectrometry and a commercial extraction system: analysis of performance, cost, and turnaround time.

PubMed

Lagacé-Wiens, Philippe R S; Adam, Heather J; Karlowsky, James A; Nichol, Kimberly A; Pang, Paulette F; Guenther, Jodi; Webb, Amanda A; Miller, Crystal; Alfa, Michelle J

2012-10-01

Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry represents a revolution in the rapid identification of bacterial and fungal pathogens in the clinical microbiology laboratory. Recently, MALDI-TOF has been applied directly to positive blood culture bottles for the rapid identification of pathogens, leading to reductions in turnaround time and potentially beneficial patient impacts. The development of a commercially available extraction kit (Bruker Sepsityper) for use with the Bruker MALDI BioTyper has facilitated the processing required for identification of pathogens directly from positive from blood cultures. We report the results of an evaluation of the accuracy, cost, and turnaround time of this method for 61 positive monomicrobial and 2 polymicrobial cultures representing 26 species. The Bruker MALDI BioTyper with the Sepsityper gave a valid (score, >1.7) identification for 85.2% of positive blood cultures with no misidentifications. The mean reduction in turnaround time to identification was 34.3 h (P < 0.0001) in the ideal situation where MALDI-TOF was used for all blood cultures and 26.5 h in a more practical setting where conventional identification or identification from subcultures was required for isolates that could not be directly identified by MALDI-TOF. Implementation of a MALDI-TOF-based identification system for direct identification of pathogens from blood cultures is expected to be associated with a marginal increase in operating costs for most laboratories. However, the use of MALDI-TOF for direct identification is accurate and should result in reduced turnaround time to identification.

Insecticide-treated clothes for the control of vector-borne diseases: a review on effectiveness and safety.

PubMed

Banks, S D; Murray, N; Wilder-Smith, A; Logan, J G

2014-08-01

Insecticide-treated clothing has been used for many years by the military and in recreational activities as personal protection against bites from a variety of arthropods including ticks, chigger mites, sandflies and mosquitoes. Permethrin is the most commonly used active ingredient, but others, including bifenthrin, deltamethrin, cyfluthrin, DEET (N,N-diethyl-3-methylbenz-amide) and KBR3023, have also been trialled. Treatment is usually carried out by home or factory dipping. However, new microencapsulation technologies which may prolong the activity of insecticides on clothing are now available and may help to overcome the inevitable reduction in efficacy over time that occurs as a result of washing, ultraviolet light exposure, and the normal wear and tear of the fabric. The aim of this article is to review the evidence base for the use of insecticide-treated clothing for protection against bites from arthropods and its effect on arthropod-borne pathogen transmission. Although some studies do demonstrate protection against pathogen transmission, there are surprisingly few, and the level of protection provided varies according to the disease and the type of study conducted. For example, insecticide-treated clothing has been reported to give between 0% and 75% protection against malaria and between 0% and 79% protection against leishmaniasis. Studies vary in the type of treatment used, the age group of participants, the geographical location of the study, and the pathogen transmission potential. This makes it difficult to compare and assess intervention trials. Overall, there is substantial evidence that insecticide-treated clothing can provide protection against arthropod bites. Bite protection evidence suggests that insecticide-treated clothing may be useful in the prevention of pathogen transmission, but further investigations are required to accurately demonstrate transmission reduction. © 2014 The Royal Entomological Society.

Expression of the Theobroma cacao Bax-inhibitor-1 gene in tomato reduces infection by the hemibiotrophic pathogen Moniliophthora perniciosa.

PubMed

Scotton, Danielle Camargo; Azevedo, Mariana Da Silva; Sestari, Ivan; Da Silva, Jamille Santos; Souza, Lucas Anjos; Peres, Lázaro Eustáquio Pereira; Leal, Gildemberg Amorim; Figueira, Antonio

2017-10-01

Programmed cell death (PCD) plays a key role in plant responses to pathogens, determining the success of infection depending on the pathogen lifestyle and on which participant of the interaction triggers cell death. The hemibiotrophic basidiomycete Moniliophthora perniciosa is the causal agent of witches' broom disease of Theobroma cacao L. (cacao), a serious constraint for production in South America and the Caribbean. It has been hypothesized that M. perniciosa pathogenesis involves PCD, initially as a plant defence mechanism, which is diverted by the fungus to induce necrosis during the dikaryotic phase of the mycelia. Here, we evaluated whether the expression of a cacao anti-apoptotic gene would affect the incidence and severity of M. perniciosa infection using the 'Micro-Tom' (MT) tomato as a model. The cacao Bax-inhibitor-1 (TcBI-1) gene, encoding a putative basal attenuator of PCD, was constitutively expressed in MT to evaluate function. Transformants expressing TcBI-1, when treated with tunicamycin, an inducer of endoplasmic reticulum stress, showed a decrease in cell peroxidation. When the same transformants were inoculated with the necrotrophic fungal pathogens Sclerotinia sclerotiorum, Sclerotium rolfsii and Botrytis cinerea, a significant reduction in infection severity was observed, confirming TcBI-1 function. After inoculation with M. perniciosa, TcBI-1 transformant lines showed a significant reduction in disease incidence compared with MT. The overexpression of TcBI-1 appears to affect the ability of germinating spores to penetrate susceptible tissues, restoring part of the non-host resistance in MT against the S-biotype of M. perniciosa. © 2016 BSPP AND JOHN WILEY & SONS LTD.

The Genome Sequence of the Tomato-Pathogenic Actinomycete Clavibacter michiganensis subsp. michiganensis NCPPB382 Reveals a Large Island Involved in Pathogenicity▿ †

PubMed Central

Gartemann, Karl-Heinz; Abt, Birte; Bekel, Thomas; Burger, Annette; Engemann, Jutta; Flügel, Monika; Gaigalat, Lars; Goesmann, Alexander; Gräfen, Ines; Kalinowski, Jörn; Kaup, Olaf; Kirchner, Oliver; Krause, Lutz; Linke, Burkhard; McHardy, Alice; Meyer, Folker; Pohle, Sandra; Rückert, Christian; Schneiker, Susanne; Zellermann, Eva-Maria; Pühler, Alfred; Eichenlaub, Rudolf; Kaiser, Olaf; Bartels, Daniela

2008-01-01

Clavibacter michiganensis subsp. michiganensis is a plant-pathogenic actinomycete that causes bacterial wilt and canker of tomato. The nucleotide sequence of the genome of strain NCPPB382 was determined. The chromosome is circular, consists of 3.298 Mb, and has a high G+C content (72.6%). Annotation revealed 3,080 putative protein-encoding sequences; only 26 pseudogenes were detected. Two rrn operons, 45 tRNAs, and three small stable RNA genes were found. The two circular plasmids, pCM1 (27.4 kbp) and pCM2 (70.0 kbp), which carry pathogenicity genes and thus are essential for virulence, have lower G+C contents (66.5 and 67.6%, respectively). In contrast to the genome of the closely related organism Clavibacter michiganensis subsp. sepedonicus, the genome of C. michiganensis subsp. michiganensis lacks complete insertion elements and transposons. The 129-kb chp/tomA region with a low G+C content near the chromosomal origin of replication was shown to be necessary for pathogenicity. This region contains numerous genes encoding proteins involved in uptake and metabolism of sugars and several serine proteases. There is evidence that single genes located in this region, especially genes encoding serine proteases, are required for efficient colonization of the host. Although C. michiganensis subsp. michiganensis grows mainly in the xylem of tomato plants, no evidence for pronounced genome reduction was found. C. michiganensis subsp. michiganensis seems to have as many transporters and regulators as typical soil-inhabiting bacteria. However, the apparent lack of a sulfate reduction pathway, which makes C. michiganensis subsp. michiganensis dependent on reduced sulfur compounds for growth, is probably the reason for the poor survival of C. michiganensis subsp. michiganensis in soil. PMID:18192381

Melanin-templated rapid synthesis of silver nanostructures

PubMed Central

2014-01-01

Background As a potent antimicrobial agent, silver nanostructures have been used in nanosensors and nanomaterial-based assays for the detection of food relevant analytes such as organic molecules, aroma, chemical contaminants, gases and food borne pathogens. In addition silver based nanocomposites act as an antimicrobial for food packaging materials. In this prospective, the food grade melanin pigment extracted from sponge associated actinobacterium Nocardiopsis alba MSA10 and melanin mediated synthesis of silver nanostructures were studied. Based on the present findings, antimicrobial nanostructures can be developed against food pathogens for food industrial applications. Results Briefly, the sponge associated actinobacterium N. alba MSA10 was screened and fermentation conditions were optimized for the production of melanin pigment. The Plackett-Burman design followed by a Box-Behnken design was developed to optimize the concentration of most significant factors for improved melanin yield. The antioxidant potential, reductive capabilities and physiochemical properties of Nocardiopsis melanin was characterized. The optimum production of melanin was attained with pH 7.5, temperature 35°C, salinity 2.5%, sucrose 25 g/L and tyrosine 12.5 g/L under submerged fermentation conditions. A highest melanin production of 3.4 mg/ml was reached with the optimization using Box-Behnken design. The purified melanin showed rapid reduction and stabilization of silver nanostructures. The melanin mediated process produced uniform and stable silver nanostructures with broad spectrum antimicrobial activity against food pathogens. Conclusions The melanin pigment produced by N. alba MSA10 can be used for environmentally benign synthesis of silver nanostructures and can be useful for food packaging materials. The characteristics of broad spectrum of activity against food pathogens of silver nanostructures gives an insight for their potential applicability in incorporation of food packaging materials and antimicrobials for stored fruits and foods. PMID:24885756

Omics Approaches for the Engineering of Pathogen Resistant Plants.

PubMed

Gomez-Casati, Diego F; Pagani, María A; Busi, María V; Bhadauria, Vijai

2016-01-01

The attack of different pathogens, such as bacteria, fungi and viruses has a negative impact on crop production. In counter such attacks, plants have developed different strategies involving the modification of gene expression, activation of several metabolic pathways and post-translational modification of proteins, which culminate into the accumulation of primary and secondary metabolites implicated in plant defense responses. The recent advancement in omics techniques allows the increase coverage of plants transcriptomes, proteomes and metabolomes during pathogen attack, and the modulation of the response after the infection. Omics techniques also allow us to learn more about the biological cycle of the pathogens in addition to the identification of novel virulence factors in pathogens and their host targets. Both approaches become important to decipher the mechanism underlying pathogen attacks and to develop strategies for improving disease-resistant plants. In this review, we summarize some of the contribution of genomics, transcriptomics, proteomics, metabolomics and metallomics in devising the strategies to obtain plants with increased resistance to pathogens. These approaches constitute important research tools in the development of new technologies for the protection against diseases and increase plant production.

Composition of growth factors and cytokines in lysates obtained from fresh versus stored pathogen-inactivated platelet units.

PubMed

Sellberg, Felix; Berglund, Erik; Ronaghi, Martin; Strandberg, Gabriel; Löf, Helena; Sommar, Pehr; Lubenow, Norbert; Knutson, Folke; Berglund, David

2016-12-01

Platelet lysate is a readily available source of growth factors, and other mediators, which has been used in a variety of clinical applications. However, the product remains poorly standardized and the present investigation evaluates the composition of platelet lysate obtained from either fresh or stored pathogen-inactivated platelet units. Platelet pooled units (n = 10) were obtained from healthy blood donors and tested according to standard procedures. All units were pathogen inactivated using amotosalen hydrochloride and UVA exposure. Platelet lysate was subsequently produced at two separate time-points, either from fresh platelet units or after 5 days of storage, by repeated freeze-thaw cycles. The following mediators were determined at each time-point: EGF, FGF-2, VEGF, IGF-1, PDGF-AB/BB, BMP-2, PF4, TGF-β isoform 1, IL-1β, IL-2, IL-6, IL-10, IL-12p70, 1L-17A, TNF-α, and IFN-γ. The concentration of growth factors and cytokines was affected by time in storage. Notably, TGF-β, PDGF-AB/BB, and PF4 showed an increase of 27.2% (p < 0.0001), 29.5% (p = 0.04) and 8.2% (p = 0.0004), respectively. A decrease was seen in the levels of IGF-1 and FGF-2 with 22% (p = 0.041) and 11% (p = 0.01), respectively. Cytokines were present only in very low concentrations and all other growth factors remained stable with time in storage. The composition of mediators in platelet lysate obtained from pathogen-inactivated platelet units differs when produced from fresh and stored platelet units, respectively. This underscores the need for further standardization and optimization of this important product, which potentially may influence the clinical effects. Copyright © 2016. Published by Elsevier Ltd.

Mitigation of Irrigation Water Using Zero-Valent Iron Treatment

USDA-ARS?s Scientific Manuscript database

Significant problems have occurred in the U.S. with regard to the contamination of produce by pathogenic bacteria. Minimally processed produce lacks the processing and preparation hurdles, such as cooking, to aid in reduction or elimination of the occasional and incidental contamination that can le...

Water Quality Modeling in the Dead End Sections of Drinking Water Distribution Networks

EPA Science Inventory

Dead-end sections of drinking water distribution networks are known to be problematic zones in terms of water quality degradation. Extended residence time due to water stagnation leads to rapid reduction of disinfectant residuals allowing the regrowth of microbial pathogens. Wate...

40 CFR 503.15 - Operational standards-pathogens and vector attraction reduction.

Code of Federal Regulations, 2010 CFR

2010-07-01

... requirements in § 503.32(a) shall be met when sewage sludge is sold or given away in a bag or other container...) shall be met when sewage sludge is sold or given away in a bag or other container for application to the...

21 CFR 120.20 - General.

Code of Federal Regulations, 2010 CFR

2010-04-01

... 21 Food and Drugs 2 2010-04-01 2010-04-01 false General. 120.20 Section 120.20 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN CONSUMPTION HAZARD ANALYSIS AND CRITICAL CONTROL POINT (HACCP) SYSTEMS Pathogen Reduction § 120.20 General...

21 CFR 120.20 - General.

Code of Federal Regulations, 2014 CFR

2014-04-01

... 21 Food and Drugs 2 2014-04-01 2014-04-01 false General. 120.20 Section 120.20 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN CONSUMPTION HAZARD ANALYSIS AND CRITICAL CONTROL POINT (HACCP) SYSTEMS Pathogen Reduction § 120.20 General...

21 CFR 120.20 - General.

Code of Federal Regulations, 2013 CFR

2013-04-01

... 21 Food and Drugs 2 2013-04-01 2013-04-01 false General. 120.20 Section 120.20 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN CONSUMPTION HAZARD ANALYSIS AND CRITICAL CONTROL POINT (HACCP) SYSTEMS Pathogen Reduction § 120.20 General...

21 CFR 120.20 - General.

Code of Federal Regulations, 2011 CFR

2011-04-01

... 21 Food and Drugs 2 2011-04-01 2011-04-01 false General. 120.20 Section 120.20 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN CONSUMPTION HAZARD ANALYSIS AND CRITICAL CONTROL POINT (HACCP) SYSTEMS Pathogen Reduction § 120.20 General...

21 CFR 120.20 - General.

Code of Federal Regulations, 2012 CFR

2012-04-01

... 21 Food and Drugs 2 2012-04-01 2012-04-01 false General. 120.20 Section 120.20 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN CONSUMPTION HAZARD ANALYSIS AND CRITICAL CONTROL POINT (HACCP) SYSTEMS Pathogen Reduction § 120.20 General...

Salmonella in lymph nodes of cull and fed cattle at harvest

USDA-ARS?s Scientific Manuscript database

Category: Pre-harvest pathogen reduction Objective: To evaluate the potential association between Salmonella enterica burden within bovine subiliac lymph nodes (LNs), animal type, season, and region of harvest. Methods: Bovine LNs (n = 4,764) were collected from 12 abattoirs, 8 that primarily h...

REDUCTION IN DBP PRECURSORS AND PATHOGENS DURING RIVERBANK FILTRATION AT THREE MIDWESTERN DRINKING WATER UTILITIES

EPA Science Inventory

Riverbank filtrtion (RBF) is a process that subjects river water to ground passage prior to its use as a drinking water supply. European expereince with RBF demonstrate that during infiltration and underground transport, processes such as filtration, sorption, and biodegradation...

EFFECTS OF STORAGE ON STABILITY AND PATHOGEN REDUCTION IN BIOSOLIDS

EPA Science Inventory

Storage can be an effective means of stabilizing small quantities of wastewater sludge. This paper summarizes the performance of two laboratory-scale sludge storage units and that of four full-scale tanks sampled at four treatment facilities in eastern Nebraska. The bench-scale u...

Advances in the genetic improvement of Prunus domestica utilizing genetic engineering

USDA-ARS?s Scientific Manuscript database

Plum producers world-wide are facing multiple challenges including climate change, reductions in available labor, the need for reduced chemical inputs, the spread of native and exotic pests and pathogens, and consumer demands for improved fruit quality and health benefits. Meeting these challenges ...

The Comprehensive Phytopathogen Genomics Resource: a web-based resource for data-mining plant pathogen genomes.

PubMed

Hamilton, John P; Neeno-Eckwall, Eric C; Adhikari, Bishwo N; Perna, Nicole T; Tisserat, Ned; Leach, Jan E; Lévesque, C André; Buell, C Robin

2011-01-01

The Comprehensive Phytopathogen Genomics Resource (CPGR) provides a web-based portal for plant pathologists and diagnosticians to view the genome and trancriptome sequence status of 806 bacterial, fungal, oomycete, nematode, viral and viroid plant pathogens. Tools are available to search and analyze annotated genome sequences of 74 bacterial, fungal and oomycete pathogens. Oomycete and fungal genomes are obtained directly from GenBank, whereas bacterial genome sequences are downloaded from the A Systematic Annotation Package (ASAP) database that provides curation of genomes using comparative approaches. Curated lists of bacterial genes relevant to pathogenicity and avirulence are also provided. The Plant Pathogen Transcript Assemblies Database provides annotated assemblies of the transcribed regions of 82 eukaryotic genomes from publicly available single pass Expressed Sequence Tags. Data-mining tools are provided along with tools to create candidate diagnostic markers, an emerging use for genomic sequence data in plant pathology. The Plant Pathogen Ribosomal DNA (rDNA) database is a resource for pathogens that lack genome or transcriptome data sets and contains 131 755 rDNA sequences from GenBank for 17 613 species identified as plant pathogens and related genera. Database URL: http://cpgr.plantbiology.msu.edu.

Molecular phylogeny, pathogenicity and toxigenicity of Fusarium oxysporum f. sp. lycopersici

PubMed Central

Nirmaladevi, D.; Venkataramana, M.; Srivastava, Rakesh K.; Uppalapati, S. R.; Gupta, Vijai Kumar; Yli-Mattila, T.; Clement Tsui, K. M.; Srinivas, C.; Niranjana, S. R.; Chandra, Nayaka S.

2016-01-01

The present study aimed at the molecular characterization of pathogenic and non pathogenic F. oxysporum f. sp. lycopersici strains isolated from tomato. The causal agent isolated from symptomatic plants and soil samples was identified based on morphological and molecular analyses. Pathogenicity testing of 69 strains on five susceptible tomato varieties showed 45% of the strains were highly virulent and 30% were moderately virulent. Molecular analysis based on the fingerprints obtained through ISSR indicated the presence of wide genetic diversity among the strains. Phylogenetic analysis based on ITS sequences showed the presence of at least four evolutionary lineages of the pathogen. The clustering of F. oxysporum with non pathogenic isolates and with the members of other formae speciales indicated polyphyletic origin of F. oxysporum f. sp. lycopersici. Further analysis revealed intraspecies variability and nucleotide insertions or deletions in the ITS region among the strains in the study and the observed variations were found to be clade specific. The high genetic diversity in the pathogen population demands for development of effective resistance breeding programs in tomato. Among the pathogenic strains tested, toxigenic strains harbored the Fum1 gene clearly indicating that the strains infecting tomato crops have the potential to produce Fumonisin. PMID:26883288

Trends in antibiotic resistance over time among pathogens from Canadian hospitals: results of the CANWARD study 2007-11.

PubMed

Lagacé-Wiens, Philippe R S; Adam, Heather J; Low, Donald E; Blondeau, Joseph M; Baxter, Melanie R; Denisuik, Andrew J; Nichol, Kimberly A; Walkty, Andrew; Karlowsky, James A; Mulvey, Michael R; Hoban, Daryl J; Zhanel, George G

2013-05-01

Antimicrobial resistance patterns change over time and longitudinal surveillance studies provide insight into these trends. We sought to describe the important trends in antimicrobial resistance in key pathogens across Canada to provide useful information to clinicians, policy makers and industry, to assist in optimizing antimicrobial therapy, formulary choices and drug development. We analysed longitudinal data from the CANWARD study using a multivariate regression model to control for possible effects of patient demographics on resistance, in order to assess the impact of time on antimicrobial resistance independent of other measured variables. We identified several key trends in common pathogens. In particular, we observed a statistically significant increase in the proportion of Escherichia coli isolates that were resistant to extended-spectrum cephalosporins and fluoroquinolones, an increase in the proportion of Klebsiella pneumoniae isolates that were resistant to extended-spectrum cephalosporins, a reduction in the proportion of Staphylococcus aureus that were methicillin, clindamycin and trimethoprim/sulfamethoxazole resistant, and a reduction in the proportion of Pseudomonas aeruginosa that were fluoroquinolone and gentamicin resistant. Although some of these trends, such as the dramatic increase in fluoroquinolone and cephalosporin resistance in E. coli, can be attributed to the emergence and global spread of resistant clones (e.g. ST131 E. coli), others remain unexplained. However, recognizing these trends remains important to guide changes in empirical antimicrobial therapy and drug development.

Efficacy of Combined Sous Vide-Microwave Cooking for Foodborne Pathogen Inactivation in Ready-to-Eat Chicory Stems.

PubMed

Renna, Massimiliano; Gonnella, Maria; de Candia, Silvia; Serio, Francesco; Baruzzi, Federico

2017-07-01

There is a variety of different food processing methods, which can be used to prepare ready-to-eat foods. However, the need to preserve the freshness and nutritional qualities leads to the application of mild technologies which may be insufficient to inactivate microbial pathogens. In this work, fresh chicory stems were packed under a vacuum in films, which were transparent to microwaves. These were then exposed to microwaves for different periods of time. The application of sous vide microwave cooking (SV-MW, 900 W, 2450 MHz), controlled naturally occurring mesophilic aerobic bacteria, yeasts and molds for up to 30 d when vacuum-packed vegetables were stored at 4 °C. In addition, the process lethality of the SV-MW 90 s cooking was experimentally validated. This treatment led to 6.07 ± 0.7 and 4.92 ± 0.65 log cfu/g reduction of Escherichia coli and Listeria monocytogenes inoculated over the chicory stems (100 g), respectively. With an initial load of 9 log cfu/g for both pathogens, less than 10 cfu/g of surviving cells were found after 90 s cooking. This shows that short-time microwave cooking can be used to effectively pasteurize vacuum-packed chicory stems, achieving >5 log cfu/g reduction of E. coli and L. monocytogenes. © 2017 Institute of Food Technologists®.

Tea polyphenols as an antivirulence compound Disrupt Quorum-Sensing Regulated Pathogenicity of Pseudomonas aeruginosa

PubMed Central

Yin, Honging; Deng, Yifeng; Wang, Huafu; Liu, Wugao; Zhuang, Xiyi; Chu, Weihua

2015-01-01

Green tea, a water extract of non-fermented leaves of Camellia sinensis L., is one of the nonalcoholic beverages in China. It is becoming increasingly popular worldwide, because of its refreshing, mild stimulant and medicinal properties. Here we examined the quorum sensing inhibitory potentials of tea polyphenols (TP) as antivirulence compounds both in vitro and in vivo. Biosensor assay data suggested minimum inhibitory concentrations (MICs) of TP against selected pathogens were 6.25 ~ 12.5 mg/mL. At sub-MIC, TP can specifically inhibit the production of violacein in Chromobacterium violaceum 12472 with almost 98% reduction at 3.125 mg/mL without affecting its growth rate. Moreover, TP exhibited inhibitory effects on virulence phenotypes regulated by QS in Pseudomonas aeruginosa. The total proteolytic activity, elastase, swarming motility and biofilm formation were reduced in a concentration-dependent manner. In vivo, TP treatment resulted in the reduction of P. aeruginosa pathogenicity in Caenorhabditis elegans. When its concentration was 3.125 mg/mL, the survival rate reached 63.3%. In the excision wound infection model, the wound contraction percentage in treatment groups was relatively increased and the colony-forming units (CFU) in the wound area were significantly decreased. These results suggested that TP could be developed as a novel non-antibiotic QS inhibitor without killing the bacteria but as an antivirulence compound to control bacterial infection. PMID:26548447

Influence of electron beam irradiation on growth of Phytophthora cinnamomi and its control in substrates

NASA Astrophysics Data System (ADS)

MigdaŁ, Wojciech; Orlikowski, Leszek B.; Ptaszek, Magdalena; Gryczka, Urszula

2012-08-01

Very extensive production procedure, especially in plants growing under covering, require methods, which would allow quick elimination or substantial reduction of populations of specific pathogens without affecting the growth and development of the cultivated plants. Among soil-borne pathogens, the Phytophthora species are especially dangerous for horticultural plants. In this study, irradiation with electron beam was applied to control Phytophthora cinnamomi. The influence of irradiation dose on the reduction of in vitro growth and the population density of the pathogen in treated peat and its mixture with composted pine bark (1:1), as well as the health of Chamaecyparis lawsoniana and Lavandula angustifolia plants were evaluated. Application of irradiation at a dose of 1.5 kGy completely inhibited the in vitro development of P. cinnamomi. This irradiation effect was connected with the disintegration of the hyphae and spores of the species. Irradiation of peat and its mixture with composted pine bark with 10 kGy resulted in the inhibition of stem base rot development in Ch. lawsoniana. Symptoms of the disease were not observed when the substrates were treated with 15 kGy. In the case of L. angustifolia, stem root rot was not observed on cuttings transplanted to infected peat irradiated at a dose of 10 kGy. Irradiation of the horticultural substrates did not affect plant growth.

Inactivation of Escherichia coli inoculated onto fresh-cut chopped cabbage using electron-beam processing.

PubMed

Grasso, Elizabeth M; Uribe-Rendon, Roberto M; Lee, Ken

2011-01-01

During the past decade there were more than 50 reported outbreaks involving leafy green vegetables contaminated with foodborne pathogens. Leafy greens, including cabbage, are fresh foods rarely heated before consumption, which enables foodborne illness. The need for improved safety of fresh food drives the demand for nonthermal food processes to decrease the risk of pathogens while maintaining fresh quality. This study examines the efficacy of electron-beam (e-beam) irradiation in decreasing indigenous microflora on fresh-cut cabbage and determines the optimal dosage to pasteurize fresh-cut cabbage inoculated with Escherichia coli K-12. Fresh-cut cabbage (100 g) was inoculated with ∼8 log E. coli K-12 and e-beam irradiated at doses of 0, 1.0, 2.3, or 4.0 kGy. At 2.3 kGy there was <1.0 log indigenous microflora remaining, indicating greater than a 4.0-log reduction by e-beam. At a 4.0-kGy dose there was >7-log reduction of E. coli K-12 in the fresh-cut cabbage. The D(10)-value for E. coli K-12 in fresh-cut cabbage was 0.564 kGy. E-beam irradiation is thus a viable nonthermal treatment that extends the shelf life and increases the safety of fresh cabbage by reducing or eliminating indigenous microflora and unwanted pathogens.

Subgingival bacterial recolonization after scaling and root planing in smokers with chronic periodontitis.

PubMed

Feres, M; Bernal, Mac; Matarazzo, F; Faveri, M; Duarte, P M; Figueiredo, L C

2015-06-01

The aim of this study was to compare subgingival bacterial recolonization patterns after scaling and root planing in current smokers and non-smokers. 15 smokers and 15 non-smokers with chronic periodontitis received scaling and root planing in six visits lasting one hour each, over a period of 21 days. Clinical monitoring was performed at baseline and 180 days, and microbiological monitoring was performed at baseline, immediately after scaling and root planing (Day 0) and at 42, 63 and 180 days post-therapy. Subgingival plaque samples were analysed by checkerboard DNA-DNA hybridization. An improvement in clinical condition was observed for smokers and non-smokers; however, non-smokers showed a greater reduction in mean clinical attachment level in intermediate sites in comparison with smokers (p < 0.05). At Day 0, there was a significant reduction in the mean counts of the three pathogens from the red complex, Eubacterium nodatum and Parvimonas micra only in non-smokers (p < 0.05). There was a significant increase in the proportion of host-compatible species in non-smokers and smokers from baseline to 180 days post-therapy (p < 0.05). However, a significant decrease in the pathogenic species was observed only in non-smokers. Smokers were more susceptible to the re-establishment of a pathogenic subgingival biofilm than non-smokers. © 2015 Australian Dental Association.

Genome Reduction for Niche Association in Campylobacter Hepaticus, A Cause of Spotty Liver Disease in Poultry.

PubMed

Petrovska, Liljana; Tang, Yue; Jansen van Rensburg, Melissa J; Cawthraw, Shaun; Nunez, Javier; Sheppard, Samuel K; Ellis, Richard J; Whatmore, Adrian M; Crawshaw, Tim R; Irvine, Richard M

2017-01-01

The term "spotty liver disease" (SLD) has been used since the late 1990s for a condition seen in the UK and Australia that primarily affects free range laying hens around peak lay, causing acute mortality and a fall in egg production. A novel thermophilic SLD-associated Campylobacter was reported in the United Kingdom (UK) in 2015. Subsequently, similar isolates occurring in Australia were formally described as a new species, Campylobacter hepaticus . We describe the comparative genomics of 10 C. hepaticus isolates recovered from 5 geographically distinct poultry holdings in the UK between 2010 and 2012. Hierarchical gene-by-gene analyses of the study isolates and representatives of 24 known Campylobacter species indicated that C. hepaticus is most closely related to the major pathogens Campylobacter jejuni and Campylobacter coli . We observed low levels of within-farm variation, even between isolates collected over almost 3 years. With respect to C. hepaticus genome features, we noted that the study isolates had a ~140 Kb reduction in genome size, ~144 fewer genes, and a lower GC content compared to C. jejuni . The most notable reduction was in the subsystem containing genes for iron acquisition and metabolism, supported by reduced growth of C. hepaticus in an iron depletion assay. Genome reduction is common among many pathogens and in C. hepaticus has likely been driven at least in part by specialization following the occupation of a new niche, the chicken liver.

Galleria mellonella as an in vivo model for assessing the protective activity of probiotics against gastrointestinal bacterial pathogens.

PubMed

Scalfaro, Concetta; Iacobino, Angelo; Nardis, Chiara; Franciosa, Giovanna

2017-04-01

The antagonistic activity against gastrointestinal bacterial pathogens is an important property of probiotic bacteria and a desirable feature for pre-selection of novel strains with probiotic potential. Pre-screening of candidate probiotics for antibacterial activity should be based on in vitro and in vivo tests. This study investigated whether the protective activity of probiotic bacteria against gastrointestinal bacterial pathogens can be evaluated using Galleria mellonella larvae as an in vivo model. Larvae were pre-inoculated with either of two widely used probiotic bacteria, Lactobacillus rhamnosus GG or Clostridium butyricum Miyairi 588, and then challenged with Salmonella enterica Typhimurium, enteropathogenic Escherichia coli or Listeria monocytogenes. Survival rates increased in the probiotic pretreated larvae compared with control larvae inoculated with pathogens only. The hemocyte density increased as well in the probiotic pretreated larvae, indicating that both probiotics induce an immune response in the larvae. The antibacterial activity of probiotics against the pathogens was also assayed by an in vitro agar spot test: results were partially consistent with those obtained by the G. mellonella protection assay. The results obtained, as a whole, suggest that G. mellonella larvae are a potentially useful in vivo model that can complement in vitro assays for pre-screening of candidate probiotics. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Assessing Antibacterial Potential of Components of Phyllomedusa distincta Skin and its Associated Dermal Microbiota.

PubMed

Brito de Assis, Ananda; Dos Santos, Cristiane; Dutra, Flávia Pereira; de Oliveira Motta, Ailla; Costa, Flávio Silva; Navas, Carlos Arturo; Magalhães, Beatriz Simas; Barreto, Cristine Chaves

2016-02-01

The granular glands of anuran skin secrete an array of bioactive molecules that protect a frog against pathogens and predators. The skin also harbors a microbial community. Although there is evidence to suggest that the microbiota complement the innate immune defense systems against pathogen infection, the effect of the frog bioactive molecules on its resident microbiota has not yet been fully investigated. In the present study, the skin microbiota of Phyllomedusa distincta obtained from two different geographical areas was evaluated with molecular and culture-based approaches. The antagonistic effects exhibited by the host's microbiota and by a novel dermaseptin peptide isolated from P. distincta skin were investigated. Four isolated bacterial colonies displayed antimicrobial activity against known frog pathogens. Our results were consistent with the hypothesis that microbiota from P. distincta may interact with pathogenic microorganisms to protect a frog's health. On the other hand, the novel dermaseptin peptide exhibited an antimicrobial effect on pathogens as well as on some of the bacteria obtained from the skin microbiota. The richness of bacteria on P. distincta skin was further investigated by 16S rRNA gene clone libraries, which revealed that the family Enterobacteriaceae was prevalent, but a high variability at the species level was observed among individual frogs. Differences observed on the microbiota of frogs from contrasting habitats indicated an influence of the environment on the structure of the skin microbiota of P. distincta.

Suitable areas of Phakopsora pachyrhizi, Spodoptera exigua, and their host plant Phaseolus vulgaris are projected to reduce and shift due to climate change

NASA Astrophysics Data System (ADS)

Ramirez-Cabral, Nadiezhda Yakovleva Zitz; Kumar, Lalit; Shabani, Farzin

2018-01-01

Worldwide, crop pests (CPs) such as pathogens and insects affect agricultural production detrimentally. Species distribution models can be used for projecting current and future suitability of CPs and host crop localities. Our study overlays the distribution of two CPs (Asian soybean rust and beet armyworm) and common bean, a potential host of them, in order to determine their current and future levels of coexistence. This kind of modeling approach has rarely been performed previously in climate change studies. The soybean rust and beet armyworm model projections herein show a reduction of the worldwide area with high and medium suitability of both CPs and a movement of them away from the Equator, in 2100 more pronounced than in 2050. Most likely, heat and dry stress will be responsible for these changes. Heat and dry stress will greatly reduce and shift the future suitable cultivation area of common bean as well, in a similar manner. The most relevant findings of this study were the reduction of the suitable areas for the CPs, the reduction of the risk under future scenarios, and the similarity of trends for the CPs and host. The current results highlight the relation between and the coevolution of host and pathogens.

[Infantile hypophosphatasia caused by a novel compound heterozygous mutation: a case report and pedigree analysis].

PubMed

Li, Deng-Feng; Lan, Dan; Zhong, Jing-Zi; Dewan, Roma Kajal; Xie, Yan-Shu; Yang, Ying

2017-05-01

This article reported the clinical features of one child with infantile hypophosphatasia (HPP) and his pedigree information. The proband was a 5-month-old boy with multiple skeletal dysplasia (koilosternia, bending deformity of both radii, and knock-knee deformity of both knees), feeding difficulty, reduction in body weight, developmental delay, recurrent pneumonia and respiratory failure, and a significant reduction in blood alkaline phosphatase. Among his parents, sister, uncle, and aunt (other family members did not cooperate with us in the examination), his parents and aunt had a slight reduction in alkaline phosphatase and his aunt had scoliosis; there were no other clinical phenotypes or abnormal laboratory testing results. His ALPL gene mutation came from c.228delG mutation in his mother and c.407G>A compound heterozygous mutation in his father. His aunt carried c.228delG mutation. The c.407G>A mutation had been reported as the pathogenic mutation of HPP, and c.228delG mutation was a novel pathogenic mutation. Hypophosphatasia is caused by ALPL gene mutation, and ALPL gene detection is an effective diagnostic method. This study expands the mutation spectrum of ALPL gene and provides a theoretical basis for genetic diagnosis of this disease.

Male-Specific Coliphages as Indicators of Thermal Inactivation of Pathogens in Biosolids

PubMed Central

Nappier, Sharon P.; Aitken, Michael D.; Sobsey, Mark D.

2006-01-01

Male-specific (F+) coliphages have been proposed as a candidate indicator of fecal contamination and of virus reduction in waste treatment. However, in this and earlier work with a laboratory thermophilic anaerobic digester, a heat-resistant fraction of F+ coliphage populations indigenous to municipal wastewater and sludge was evident. We therefore isolated coliphages from municipal wastewater sludge and from biosolid samples after thermophilic anaerobic digestion to evaluate the susceptibility of specific groups to thermal inactivation. Similar numbers of F+ DNA and F+ RNA coliphages were found in untreated sludge, but the majority of isolates in digested biosolids were group I F+ RNA phages. Separate experiments on individual isolates at 53°C confirmed the apparent heat resistance of group I F+ RNA coliphages as well as the susceptibility of group III F+ RNA coliphages. Although few F+ DNA coliphages were recovered from the treated biosolid samples, thermal inactivation experiments indicated heat resistance similar to that of group I F+ RNA phages. Hence, F+ DNA coliphage reductions during thermophilic anaerobic digestion are probably related to mechanisms other than thermal inactivation. Further studies should focus on the group III F+ RNA coliphages as potential indicators of reductions of heat-resistant pathogens in thermal processes for sludge treatment. PMID:16597945

The membrane as the gatekeeper of infection: Cholesterol in host-pathogen interaction.

PubMed

Kumar, G Aditya; Jafurulla, Md; Chattopadhyay, Amitabha

2016-09-01

The cellular plasma membrane serves as a portal for the entry of intracellular pathogens. An essential step for an intracellular pathogen to gain entry into a host cell therefore is to be able to cross the cell membrane. In this review, we highlight the role of host membrane cholesterol in regulating the entry of intracellular pathogens using insights obtained from work on the interaction of Leishmania and Mycobacterium with host cells. The entry of these pathogens is known to be dependent on host membrane cholesterol. Importantly, pathogen entry is inhibited either upon depletion (or complexation), or enrichment of membrane cholesterol. In other words, an optimum level of host membrane cholesterol is necessary for efficient infection by pathogens. In this overall context, we propose a general mechanism, based on cholesterol-induced conformational changes, involving cholesterol binding sites in host cell surface receptors that are implicated in this process. A therapeutic strategy targeting modulation of membrane cholesterol would have the advantage of avoiding the commonly encountered problem of drug resistance in tackling infection by intracellular pathogens. Insights into the role of host membrane cholesterol in pathogen entry would be instrumental in the development of novel therapeutic strategies to effectively tackle intracellular pathogenesis. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

Transformation of Sclerotinia Sclerotiorum with the Green Fluorescent Protein Gene and Fluorescence of Hyphae in Four Inoculated Hosts

USDA-ARS?s Scientific Manuscript database

Sclerotinia sclerotiorum is an important pathogen of a wide variety of crops. To obtain a genetic marker to observe and study the interaction of the pathogen with its hosts, isolates ND30 and ND21 were transformed using pCT74 and gGFP constructs both containing genes for the green fluorescent protei...

Two single mutations in the fusion protein of Newcastle disease virus confer hemagglutinin-neuraminidase independent fusion promotion and attenuate the pathogenicity in chickens

USDA-ARS?s Scientific Manuscript database

The fusion (F) protein of Newcastle disease virus (NDV) plays an important role in viral infection and pathogenicity through mediating membrane fusion between the virion and host cells in the presence of the hemagglutinin-neuraminidase (HN). Previously, we obtained a velogenic NDV genotype VII muta...

A new laboratory cultivation of Paramecium bursaria using non-pathogenic bacteria strains.

PubMed

Bator, Tomasz

2010-01-01

In most studies dealing with the laboratory cultivation of paramecia (Paramecium bursaria), Klebsiella pneumoniae bacteria are used to inoculate the medium. However, Klebsiella pneumoniae is a typical pathogen, and its use is always associated with a risk of infection. The aim of the present research was to examine non-pathogenic bacteria strains as components of the medium for Paramecium bursaria. The paramecia were incubated on lettuce infusions bacterized with different bacteria strains: Bacillus subtilis DSM 10, Bacillus megaterium DSM 32, Escherichia coli DSM 498, Micrococcus luteus DSM 348. A strain derived from the natural habitat of Paramecium bursaria was used as the control one. Experiments were conducted under constant light and in the dark. Paramecia cells were counted under a stereomicroscope on consecutive days of incubation. The obtained results show that the most intensive growth of Paramecium bursaria occurs in the presence of Escherichia coli DSM 498. The use of this strain as a component of the medium allows one to obtain a high number of ciliates regardless of the light conditions. It can be concluded that the Paramecium bursaria cultivation procedure can be modified by using the non-pathogenic bacteria strain Escherichia coli DSM 498 instead of Klebsiella pneumoniae.

Research advances on microbial genetics in China in 2015.

PubMed

Xie, Jian-ping; Han, Yu-bo; Liu, Gang; Bai, Lin-quan

2016-09-01

In 2015, there are significant progresses in many aspects of the microbial genetics in China. To showcase the contribution of Chinese scientists in microbial genetics, this review surveys several notable progresses in microbial genetics made largely by Chinese scientists, and some key findings are highlighted. For the basic microbial genetics, the components, structures and functions of many macromolecule complexes involved in gene expression regulation have been elucidated. Moreover, the molecular basis underlying the recognition of foreign nucleic acids by microbial immune systems was unveiled. We also illustrated the biosynthetic pathways and regulators of multiple microbial compounds, novel enzyme reactions, and new mechanisms regulating microbial gene expression. And new findings were obtained in the microbial development, evolution and population genetics. For the industrial microbiology, more understanding on the molecular basis of the microbial factory has been gained. For the pathogenic microbiology, the genetic circuits of several pathogens were depicted, and significant progresses were achieved for understanding the pathogen-host interaction and revealing the genetic mechanisms underlying antimicrobial resistance, emerging pathogens and environmental microorganisms at the genomic level. In future, the genetic diversity of microbes can be used to obtain specific products, while gut microbiome is gathering momentum.

Discrete time Markov chains (DTMC) susceptible infected susceptible (SIS) epidemic model with two pathogens in two patches

NASA Astrophysics Data System (ADS)

Lismawati, Eka; Respatiwulan; Widyaningsih, Purnami

2017-06-01

The SIS epidemic model describes the pattern of disease spread with characteristics that recovered individuals can be infected more than once. The number of susceptible and infected individuals every time follows the discrete time Markov process. It can be represented by the discrete time Markov chains (DTMC) SIS. The DTMC SIS epidemic model can be developed for two pathogens in two patches. The aims of this paper are to reconstruct and to apply the DTMC SIS epidemic model with two pathogens in two patches. The model was presented as transition probabilities. The application of the model obtain that the number of susceptible individuals decreases while the number of infected individuals increases for each pathogen in each patch.

[Detection of pathogenic leptospira in the waste water and sewage sludge of large pig breeding sites].

PubMed

Mînzat, R M; Tomescu, V

1975-01-01

Sewage effluent and sludge from the purification plant of 8 large piggeries was examined for the presence of pathogenic leptospires. By using the methods of Appelman and Van Thiel it was found that 43.1% of samples of effluent were contaminated with L. pomona and O. tarassovi. Altogether 33 strains of pomona and three mixed cultures of pomona and tarassovi were obtained. The isolated strains were shown to be pathogenic by experimental infection of guinea-pigs, rabbits and pregnant and non-pregnant sows. The average period of survival of pathogenic leptospires in sewage effluent was 24 to 48 hours, with a maximum of 96 hours. Leptospires were killed within 24 hours in decanted sludge, owing to its strong acidity.

Making a protein extract from plant pathogenic fungi for gel- and LC-based proteomics.

PubMed

Fernández, Raquel González; Redondo, Inmaculada; Jorrin-Novo, Jesus V

2014-01-01

Proteomic technologies have become a successful tool to provide relevant information on fungal biology. In the case of plant pathogenic fungi, this approach would allow a deeper knowledge of the interaction and the biological cycle of the pathogen, as well as the identification of pathogenicity and virulence factors. These two elements open up new possibilities for crop disease diagnosis and environment-friendly crop protection. Phytopathogenic fungi, due to its particular cellular characteristics, can be considered as a recalcitrant biological material, which makes it difficult to obtain quality protein samples for proteomic analysis. This chapter focuses on protein extraction for gel- and LC-based proteomics with specific protocols of our current research with Botrytis cinerea.

Conceptualising the technical relationship of animal disease surveillance to intervention and mitigation as a basis for economic analysis

PubMed Central

2011-01-01

Background Surveillance and intervention are resource-using activities of strategies to mitigate the unwanted effects of disease. Resources are scarce, and allocating them to disease mitigation instead of other uses necessarily involves the loss of alternative sources of benefit to people. For society to obtain the maximum benefits from using resources, the gains from disease mitigation must be compared to the resource costs, guiding decisions made with the objective of achieving the optimal net outcome. Discussion Economics provides criteria to guide decisions aimed at optimising the net benefits from the use of scarce resources. Assessing the benefits of disease mitigation is no exception. However, the technical complexity of mitigation means that economic evaluation is not straightforward because of the technical relationship of surveillance to intervention. We argue that analysis of the magnitudes and distribution of benefits and costs for any given strategy, and hence the outcome in net terms, requires that mitigation is considered in three conceptually distinct stages. In Stage I, 'sustainment', the mitigation objective is to sustain a free or acceptable status by preventing an increase of a pathogen or eliminating it when it occurs. The role of surveillance is to document that the pathogen remains below a defined threshold, giving early warning of an increase in incidence or other significant changes in risk, and enabling early response. If a pathogen is not contained, the situation needs to be assessed as Stage II, 'investigation'. Here, surveillance obtains critical epidemiological information to decide on the appropriate intervention strategy to reduce or eradicate a disease in Stage III, 'implementation'. Stage III surveillance informs the choice, timing, and scale of interventions and documents the progress of interventions directed at prevalence reduction in the population. Summary This article originates from a research project to develop a conceptual framework and practical tool for the economic evaluation of surveillance. Exploring the technical relationship between mitigation as a source of economic value and surveillance and intervention as sources of economic cost is crucial. A framework linking the key technical relationships is proposed. Three conceptually distinct stages of mitigation are identified. Avian influenza, salmonella, and foot and mouth disease are presented to illustrate the framework. PMID:21929812

Comparison of water wash, trimming, and combined hot water and lactic acid treatments for reducing bacteria of fecal origin on beef carcasses.

PubMed

Castillo, A; Lucia, L M; Goodson, K J; Savell, J W; Acuff, G R

1998-07-01

Cleaning treatments, such as high-pressure water wash at 35 degrees C or trim, alone and combined with sanitizing treatments, such as hot water (95 degrees C at the source), warm (55 degrees C) 2% lactic acid spray, and combinations of these two sanitizing methods, were compared for their effectiveness in reducing inoculated numbers (5.0 to 6.0 log CFU/cm2) of Salmonella typhimurium, Escherichia coli O157:H7, aerobic plate counts, Enterobacteriaceae, total coliforms, thermotolerant coliforms, and generic E. coli on hot beef carcass surface areas in a model carcass spray cabinet. Log reductions in numbers of all tested organisms by water wash or trim alone were significantly smaller than the log reductions obtained by the different combined treatments. Regardless of the cleaning treatment (water wash or trim) or surface area, the range for mean log reductions by hot water was from 4.0 to > 4.8 log CFU/cm2, by lactic acid spray was from 4.6 to > 4.9 log CFU/cm2, by hot water followed by lactic acid spray was from 4.5 to > 4.9 log CFU/cm2, and by lactic acid spray followed by hot water was from 4.4 to > 4.6 log CFU/cm2, for S. typhimurium and E. coli O157:H7. Identical reductions were obtained for thermotolerant coliforms and generic E. coli. No differences in bacterial reductions were observed for different carcass surface regions. Water wash and trim treatments caused spreading of the contamination to other areas of the carcass surface while providing an overall reduction in fecal or pathogenic contamination on carcass surface areas. This relocated contamination after either water wash or trim was most effectively reduced by following with hot water and then lactic acid spray. This combined treatment yielded 0% positive samples for S. typhimurium, E. coli O157:H7, thermotolerant coliforms, and generic E. coli on areas outside the inoculated areas, whereas percent positive samples after applying other combined treatments ranged from 22 to 44% for S. typhimurium, 0 to 44% for E. coli O157:H7, and 11 to 33% for both thermotolerant coliforms and generic E. coli. From data collected in this study, it is possible to choose an effective, inexpensive treatment to reduce bacterial contamination on beef carcasses. In addition, the similar reduction rates of total coliforms, thermotolerant coliforms, or generic E. coli may be useful in identifying an indicator to verify the effectiveness of the selected treatment as a critical control point in a Hazard Analysis and Critical Control Point program.

Food contact surfaces coated with nitrogen-doped titanium dioxide: effect on Listeria monocytogenes survival under different light sources

NASA Astrophysics Data System (ADS)

Rodrigues, D.; Teixeira, P.; Tavares, C. J.; Azeredo, J.

2013-04-01

Improvement of food safety is a very important issue, and is on the basis of production and application of new/modified food contact surfaces. Titanium dioxide (TiO2) and, more recently, nitrogen-doped titanium dioxide (N-TiO2) coatings are among the possible forms to enhance food contact surfaces performance in terms of higher hygiene and easier sanitation. In this context, the present work aimed at evaluating the bactericidal activity of an N-TiO2 coating on glass and stainless steel under two different sources of visible light - fluorescent and incandescent - and ultraviolet (UV) irradiation. Listeria monocytogenes was chosen as representative of major foodborne pathogens and its survival was tested on N-TiO2 coated coupons. In terms of survival percentage, good results were obtained after exposure of coated surfaces to all light types since, apart from the value obtained after exposing glass to fluorescent light (56.3%), survival rates were always below 50%. However, no effective disinfection was obtained, given that for a disinfectant or sanitizing agent to be claimed as effective it needs to be able to promote at least a 3-log reduction of the microbial load, which was not observed for any of the experimental conditions assessed. Even so, UV irradiation was the most successful on eliminating cells on coated surfaces, since the amount of bacteria was reduced to 1.49 × 106 CFU/ml on glass and 2.37 × 107 on stainless steel. In contrast, both visible light sources had only slightly decreased the amount of viable cells, which remained in the range of 8 log CFU/ml. Hence, although some bactericidal effect was accomplished under visible light, UV was the most effective light source on promoting photocatalytic reactions on N-TiO2 coated coupons and none of the experimental conditions have reached a satisfactory disinfection level. Thus, this surface coating needs further research and improvement in order to become truly effective against foodborne pathogens and, ultimately, become a useful tool towards food safety in general.

Management of sewage sludge by composting using fermented water hyacinth.

PubMed

Tello-Andrade, A F; Jiménez-Moleón, M C; Sánchez-Galván, G

2015-10-01

The goal of the present research work was to assess the management of sewage sludge (SS) by composting using fermented water hyacinth (WHferm) as an amendment. The water hyacinth was fermented, and a higher production of volatile fatty acids (VFAs) (782.67 mg L(-1)) and soluble organic carbon (CSOL) (4788.34 mg L(-1)) was obtained using a particle size of 7 mm compared to 50 mm. For composting, four treatments (10 kg fresh weight each) were evaluated: treatment A (100 % SS + 0 % WHferm), treatment B (75 % SS + 25 % WHferm), treatment C (50 % SS + 50 % WHferm), and treatment D (25 % SS + 75 % WHferm). The WHferm added to SS, especially in treatments C (50 %) and D (75 %), increased the initial contents of organic matter (OM), organic carbon (CORG), CSOL, the C/N ratio, and the germination index (GI). The heavy metal content (HMC) (As, Cd, Cr, Cu, Hg, Ni, Pb, and Zn) at the beginning was below the maximum allowed by USEPA regulations. All of the samples were free of Salmonella sp. from the beginning. The reduction of the CORG, CSOL, total Kjeldahl nitrogen (TKN), and C/N ratio indicated the degradation of the OM by day 198. The treatments with WHferm (B, C, and D) yielded higher values of electrical conductivity, cation exchange capacity, and GI than SS at day 198. No significant differences were observed in GI among the treatments with WHferm. The fecal coliforms were eliminated (<3 MPN g(-1)) and the helminths were reduced to ≤5 eggs/2 g during the process. The competition for nutrients and the presence of suppressive fungi of the genera Penicillium, Rhizopus, Paecilomyces (penicillin producers), and Fusariella isolated from the compost may have promoted the elimination of pathogens since no thermophile temperatures were obtained. WHferm as an amendment in the composting of SS improved the characteristics of the final product, especially when it was used in proportions of 25 and 50 %. An excellent product was obtained in terms of HMC, and the product was B class in terms of pathogens.

Comprehensive In Situ Killing of Six Common Wound Pathogens With Manuka Honey Dressings Using a Modified AATCC-TM100.

PubMed

Watson, Denis; Bergquist, Stephen; Nicholson, Julie; Norrie, David H

2017-06-28

While Manuka honey in vitro is strongly antimicrobial, there have been, to the best of the authors' knowledge, no studies showing that dressings impregnated with Manuka honey can kill organisms in the dressing itself. The investigators used the American Association of Textile Chemists and Colorists' 100 test methodology to compare honey-impregnated dressings with control dressings (without honey) on the ability to kill common wound pathogens. Organisms were chosen after a review of the causal organisms found in actual wound infections over a 12-month period in a busy outpatient wound clinic. Even when the dressings were challenged daily with further inoculated organisms, > 5-log reductions were routinely noted across a range of pathogens, including multiple drug-resistant species using dressings containing Manuka honey relative to the control. The results presented herein show that when well-characterized medical-grade Manuka honey is used in dressings (ie, a minimum of 400 mg methylglyoxal/kg) these dressings can comprehensively kill common wound pathogens associated with infected wounds.

Phyto-synthesis of silver nanoscale particles using Morinda citrifolia L. and its inhibitory activity against human pathogens.

PubMed

Sathishkumar, Gnanasekar; Gobinath, Chandrakasan; Karpagam, Karuppiah; Hemamalini, Vedagiri; Premkumar, Kumpati; Sivaramakrishnan, Sivaperumal

2012-06-15

Leaf extract of Morinda citrifolia L. was assessed for the synthesis of silver nanoscale particles under different temperature and reaction time. Synthesized nanoscale (MCAgNPs) particles were confirmed by analysing the excitation of surface plasmon resonance (SPR) using UV-visible spectrophotometer at 420 nm. Further SEM, HRTEM analysis confirmed the range of particle size between 10 and 60 nm and SEAD pattern authorizes the face centered cubic (fcc) crystalline nature of the MCAgNPs. Fourier transform infrared spectrum (FTIR) of synthesized MCAgNPs confirms the presence of high amount of phenolic compounds in the plant extract which may possibly influence the reduction process and stabilization of nanoparticles. Further, inhibitory activity of MCAgNPs and plant extract were tested against human pathogens like Eschericia coli, Pseudomonas aeroginosa, Klebsiella pneumoniae, Enterobacter aerogenes, Bacillus cereus and Enterococci sp. The results indicated that the MCAgNPs showed moderate inhibitory actions against human pathogens than crude plant extract, demonstrating its antimicrobial value against pathogenic diseases. Copyright © 2012 Elsevier B.V. All rights reserved.

A Novel Therapy for Melanoma Developed in Mice: Transformation of Melanoma into Dendritic Cells with Listeria monocytogenes

PubMed Central

Bronchalo-Vicente, Lucia; Rodriguez-Del Rio, Estela; Freire, Javier; Calderon-Gonzalez, Ricardo; Frande-Cabanes, Elisabet; Gomez-Roman, Jose Javier; Fernández-Llaca, Hector; Yañez-Diaz, Sonsoles; Alvarez-Dominguez, Carmen

2015-01-01

Listeria monocytogenes is a gram-positive bacteria and human pathogen widely used in cancer immunotherapy because of its capacity to induce a specific cytotoxic T cell response in tumours. This bacterial pathogen strongly induces innate and specific immunity with the potential to overcome tumour induced tolerance and weak immunogenicity. Here, we propose a Listeria based vaccination for melanoma based in its tropism for these tumour cells and its ability to transform in vitro and in vivo melanoma cells into matured and activated dendritic cells with competent microbicidal and antigen processing abilities. This Listeria based vaccination using low doses of the pathogen caused melanoma regression by apoptosis as well as bacterial clearance. Vaccination efficacy is LLO dependent and implies the reduction of LLO-specific CD4+ T cell responses, strong stimulation of innate pro-inflammatory immune cells and a prevalence of LLO-specific CD8+ T cells involved in tumour regression and Listeria elimination. These results support the use of low doses of pathogenic Listeria as safe melanoma therapeutic vaccines that do not require antibiotics for bacterial removal. PMID:25760947

A newly evolved Drosophila Cytorace-9 shows trade-off between longevity and immune response.

PubMed

Sinam, Yoirentomba Meetei; Chatterjee, Arunita; Ranjini, Mysore S; Poojari, Adarsh; Nagarajan, Aarthi; Ramachandra, Nallur B; Nongthomba, Upendra

2016-10-01

Species with an efficient immune system would be at an advantage to evade pathogenic challenges and adapt to an ever changing ecological niche. The upkeep of immunity is a costly affair, thus trade-offs between immunity and other life history traits are expected. However, studies on the relation between immunity and life span have yielded paradoxical results. Drosophila Cytoraces, being at different stages of evolutionary divergence, provide an excellent experimental model system to study how evolving populations gain novel traits in the absence of selection. We found that in the absence of pathogenic infections, the Cytorace-9 flies lived longer than those of Cytorace-3. However, when these Cytoraces were challenged with different pathogenic microbes, the trend was opposite. After infection with pathogens, the long-lived Cytorace-9 survived worse than the short lived Cytorace-3, which can be attributed to a reduction in its immune response. This study provides evidence to support the existence of a trade-off between life span and immunity. Copyright © 2016 Elsevier B.V. All rights reserved.

Infection of Immature Ixodes scapularis (Acari: Ixodidae) by Membrane Feeding

PubMed Central

Oliver, Jonathan D.; Lynn, Geoffrey E.; Burkhardt, Nicole Y.; Price, Lisa D.; Nelson, Curtis M.; Kurtti, Timothy J.; Munderloh, Ulrike G.

2016-01-01

Abstract A reduction in the use of animals in infectious disease research is desirable for animal welfare as well as for simplification and standardization of experiments. An artificial silicone-based membrane-feeding system was adapted for complete engorgement of adult and nymphal Ixodes scapularis Say (Acari: Ixodidae), and for infecting nymphs with pathogenic, tick-borne bacteria. Six wild-type and genetically transformed strains of four species of bacteria were inoculated into sterile bovine blood and fed to ticks. Pathogens were consistently detected in replete nymphs by polymerase chain reaction. Adult ticks that ingested bacteria as nymphs were evaluated for transstadial transmission. Borrelia burgdorferi and Ehrlichia muris -like agent showed high rates of transstadial transmission to adult ticks, whereas Anaplasma phagocytophilum and Rickettsia monacensis demonstrated low rates of transstadial transmission/maintenance. Artificial membrane feeding can be used to routinely maintain nymphal and adult I. scapularis , and infect nymphs with tick-borne pathogens. PMID:26721866

Cold plasma inactivates Salmonella Stanley and Escherichia coli O157:H7 inoculated on golden delicious apples.

PubMed

Niemira, Brendan A; Sites, Joseph

2008-07-01

Cold plasma generated in a gliding arc was applied to outbreak strains of Escherichia coli O157:H7 and Salmonella Stanley on agar plates and inoculated onto the surfaces of Golden Delicious apples. This novel sanitizing technology inactivated both pathogens on agar plates, with higher flow rate (40 liters/min) observed to be more efficacious than were lower flow rates (20 liters/min), irrespective of treatment time (1 or 2 min). Golden Delicious apples were treated with various flow rates (10, 20, 30, or 40 liters/min) of cold plasma for various times (1, 2, or 3 min), applied to dried spot inoculations. All treatments resulted in significant (P < 0.05) reductions from the untreated control, with 40 liters/min more effective than were lower flow rates. Inactivation of Salmonella Stanley followed a time-dependent reduction for all flow rates. Reductions after 3 min ranged from 2.9 to 3.7 log CFU/ml, close to the limit of detection. For E. coli O157:H7, 40 liters/min gave similar reductions for all treatment times, 3.4 to 3.6 log CFU/ml. At lower flow rates, inactivation was related to exposure time, with 3 min resulting in reductions of 2.6 to 3 log CFU/ml. Temperature increase of the treated apples was related to exposure time for all flow rates. The maximum temperature of any plasma-treated apple was 50.8 degrees C (28 degrees C above ambient), after 20 liters/min for 3 min, indicating that antimicrobial effects were not the result of heat. These results indicate that cold plasma is a nonthermal process that can effectively reduce human pathogens inoculated onto fresh produce.

Identification of two novel LRP5 mutations in families with familial exudative vitreoretinopathy

PubMed Central

Fei, Ping; Zhang, Qi; Huang, Luling; Xu, Yu; Zhu, Xiong; Tai, Zhengfu; Gong, Bo; Ma, Shi; Yao, Quanyao; Li, Jing; Zhao, Peiquan

2014-01-01

Purpose To investigate the clinical features and disease-causing mutations in two Chinese families with familial exudative vitreoretinopathy (FEVR). Methods Clinical data and genomic DNA were collected for patients with FEVR. The coding exons and adjacent intronic regions of FZD4, LRP5, TSPAN12, and NDP were amplified with PCR, and the resulting amplicons were analyzed with Sanger sequencing. Wild-type and mutant LRP5 proteins were assayed for the Norrin/β-catenin pathway by luciferase reporter assays. Results Two novel heterozygous mutations in the LRP5 gene were identified in two relatives—p.A422T and p.L540P. Typical FEVR fundus change and mild reduced bone mineral density (BMD) was found in the two patients and the affected parent. In the luciferase studies, both p.A422T and p.L540P mutants displayed a significant reduction of the luciferase activity in SuperTopFlash (STF) cells in response to Norrin (87% reduction for p.A422T and 97% reduction for p.L540P). Both patients had an additional LRP5 sequence change (p.Q816P in Patient 1 from the unaffected mother and p.T852M in Patient 2 verified as a new mutation). Luciferase assay showed no reduction for p.Q816P and 94.9% reduction for the new mutation p.T852M, suggesting that p.Q816P may be not pathogenic and p.T852M may be pathogenic. Conclusions Our findings demonstrated two new novel LRP5 mutations in Chinese patients with FEVR and mild reduced BMD. They emphasize the complexity of FEVR mutations and phenotypes. PMID:24715757

Host Identity Matters in the Amphibian-Batrachochytrium dendrobatidis System: Fine-Scale Patterns of Variation in Responses to a Multi-Host Pathogen

PubMed Central

Gervasi, Stephanie; Gondhalekar, Carmen; Olson, Deanna H.; Blaustein, Andrew R.

2013-01-01

Species composition within ecological assemblages can drive disease dynamics including pathogen invasion, spread, and persistence. In multi-host pathogen systems, interspecific variation in responses to infection creates important context dependency when predicting the outcome of disease. Here, we examine the responses of three sympatric host species to a single fungal pathogen, Batrachochytrium dendrobatidis, which is associated with worldwide amphibian population declines and extinctions. Using an experimental approach, we show that amphibian species from three different genera display significant differences in patterns of pathgen-induced mortality as well as the magnitude and temporal dynamics of infection load. We exposed amphibians to one of four inoculation dose treatments at both larval and post- metamorphic stages and quantified infection load on day 8 and day 15 post-inoculation. Of the three species examined, only one (the Pacific treefrog; Pseudacris regilla) displayed “dose-dependent” responses; survival was reduced and infection load was elevated as inoculation dose was increased. We observed a reduction in survival but no differences in infection load across pathogen treatments in Cascades frogs (Rana cascadae). Western toads (Anaxyrus boreas) displayed differences in infection load but no differences in survival across pathogen treatments. Within species, responses to the pathogen varied with life history stage, and the most heavily infected species at the larval stage was different from the most heavily infected species at the post-metamorphic stage. Temporal changes in infection load were species and life history stage-specific. We show that variation in susceptibility to this multi-host pathogen is complex when viewed at a fine-scale and may be mediated through intrinsic host traits. PMID:23382904

Large-Scale Gene Disruption in Magnaporthe oryzae Identifies MC69, a Secreted Protein Required for Infection by Monocot and Dicot Fungal Pathogens

PubMed Central

Saitoh, Hiromasa; Fujisawa, Shizuko; Mitsuoka, Chikako; Ito, Akiko; Hirabuchi, Akiko; Ikeda, Kyoko; Irieda, Hiroki; Yoshino, Kae; Yoshida, Kentaro; Matsumura, Hideo; Tosa, Yukio; Win, Joe; Kamoun, Sophien; Takano, Yoshitaka; Terauchi, Ryohei

2012-01-01

To search for virulence effector genes of the rice blast fungus, Magnaporthe oryzae, we carried out a large-scale targeted disruption of genes for 78 putative secreted proteins that are expressed during the early stages of infection of M. oryzae. Disruption of the majority of genes did not affect growth, conidiation, or pathogenicity of M. oryzae. One exception was the gene MC69. The mc69 mutant showed a severe reduction in blast symptoms on rice and barley, indicating the importance of MC69 for pathogenicity of M. oryzae. The mc69 mutant did not exhibit changes in saprophytic growth and conidiation. Microscopic analysis of infection behavior in the mc69 mutant revealed that MC69 is dispensable for appressorium formation. However, mc69 mutant failed to develop invasive hyphae after appressorium formation in rice leaf sheath, indicating a critical role of MC69 in interaction with host plants. MC69 encodes a hypothetical 54 amino acids protein with a signal peptide. Live-cell imaging suggested that fluorescently labeled MC69 was not translocated into rice cytoplasm. Site-directed mutagenesis of two conserved cysteine residues (Cys36 and Cys46) in the mature MC69 impaired function of MC69 without affecting its secretion, suggesting the importance of the disulfide bond in MC69 pathogenicity function. Furthermore, deletion of the MC69 orthologous gene reduced pathogenicity of the cucumber anthracnose fungus Colletotrichum orbiculare on both cucumber and Nicotiana benthamiana leaves. We conclude that MC69 is a secreted pathogenicity protein commonly required for infection of two different plant pathogenic fungi, M. oryzae and C. orbiculare pathogenic on monocot and dicot plants, respectively. PMID:22589729

The effect of composting on the persistence of four ionophores in dairy manure and poultry litter

USDA-ARS?s Scientific Manuscript database

Manure composting is a well-described approach for stabilization of nutrients and reduction of pathogens and odors. Although composting studies have shown that thermophilic temperatures and aerobic conditions can increase removal rates of selected antibiotics, comparable information is lacking for ...

Evaluation of wild Juglans species for crown gall resistance

USDA-ARS?s Scientific Manuscript database

Paradox, the most widely used rootstock in CA walnut production, is highly susceptible to the causal agent of crown gall (CG) Agrobacterium tumefaciens. The bacterial pathogen induces the formation of large tumors around the crown of the tree resulting in a reduction in both vigor and yield. If left...

21 CFR 120.24 - Process controls.

Code of Federal Regulations, 2010 CFR

2010-04-01

... 21 Food and Drugs 2 2010-04-01 2010-04-01 false Process controls. 120.24 Section 120.24 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN CONSUMPTION HAZARD ANALYSIS AND CRITICAL CONTROL POINT (HACCP) SYSTEMS Pathogen Reduction § 120.24 Process...

Improved annotation of the insect vector of citrus greening disease: Biocuration by a diverse genomics community

USDA-ARS?s Scientific Manuscript database

The Asian citrus psyllid (Diaphorina citri Kuwayama) is the insect vector of the bacterium Candidatus Liberibacter asiaticus (CLas), the pathogen associated with citrus Huanglongbing (HLB, citrus greening). HLB threatens citrus production worldwide. Suppression or reduction of the insect vector usin...

Reduction in fecundity and shifts in cellular processes by a native virus on an invasive insect

USDA-ARS?s Scientific Manuscript database

Pathogens and their vectors have co-evolutionary histories that are intricately intertwined with their ecologies, environments and genetic interactions. The majority of non-persistently transmitted plant viruses are transmitted by aphid species. One important aphid vector in soybean-growing regions ...

75 FR 37376 - Notice of Request for Revision of a Currently Approved Information Collection (Pathogen Reduction...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-06-29

... Control Point (HACCP) Systems requirements because OMB approval will expire on December 31, 2010, and to... collection. FSIS has established requirements applicable to meat and poultry establishments designed to... by slaughter establishments to verify the adequacy of the establishment's process controls for the...

Water Quality Modeling in the Dead End Sections of Drinking Water Distribution Networks -journal article

EPA Science Inventory

Dead-end sections of drinking water distribution networks are known to be problematic zones in terms of water quality degradation. Extended residence time due to water stagnation leads to rapid reduction of disinfectant residuals allowing the regrowth of microbial pathogens. Wate...

Transcriptome profiling of soybean (Glycine max) roots challenged with pathogenic and non-pathogenic isolates of Fusarium oxysporum.

PubMed

Lanubile, Alessandra; Muppirala, Usha K; Severin, Andrew J; Marocco, Adriano; Munkvold, Gary P

2015-12-21

Fusarium oxysporum is one of the most common fungal pathogens causing soybean root rot and seedling blight in U.S.A. In a recent study, significant variation in aggressiveness was observed among isolates of F. oxysporum collected from roots in Iowa, ranging from highly pathogenic to weakly or non-pathogenic isolates. We used RNA-seq analysis to investigate the molecular aspects of the interactions of a partially resistant soybean genotype with non-pathogenic/pathogenic isolates of F. oxysporum at 72 and 96 h post inoculation (hpi). Markedly different gene expression profiles were observed in response to the two isolates. A peak of highly differentially expressed genes (HDEGs) was triggered at 72 hpi in soybean roots and the number of HDEGs was about eight times higher in response to the pathogenic isolate compared to the non-pathogenic one (1,659 vs. 203 HDEGs, respectively). Furthermore, the magnitude of induction was much greater in response to the pathogenic isolate. This response included a stronger activation of defense-related genes, transcription factors, and genes involved in ethylene biosynthesis, secondary and sugar metabolism. The obtained data provide an important insight into the transcriptional responses of soybean-F. oxysporum interactions and illustrate the more drastic changes in the host transcriptome in response to the pathogenic isolate. These results may be useful in the developing new methods of broadening resistance of soybean to F. oxysporum, including the over-expression of key soybean genes.

Compressed sodium chloride as a fast-acting antimicrobial surface: results of a pilot study.

PubMed

Whitlock, B D; Smith, S W

2016-10-01

Antimicrobial surfaces are currently being studied as an aid to reduce transmission of pathogens leading to healthcare-associated infections (HAIs). Among the most harmful and costly pathogens that cause HAIs is meticillin-resistant Staphylococcus aureus (MRSA). Currently available and previously investigated antimicrobial surface technologies that are effective against MRSA (e.g. copper alloy surfaces) take 30min to several hours to achieve significant reduction. This article presents a new antimicrobial surface technology made of compressed sodium chloride that reduces MRSA 20-30 times faster than copper alloy surfaces. Copyright © 2016 The Healthcare Infection Society. Published by Elsevier Ltd. All rights reserved.

No hormone to rule them all: Interactions of plant hormones during the responses of plants to pathogens.

PubMed

Shigenaga, Alexandra M; Argueso, Cristiana T

2016-08-01

Plant hormones are essential regulators of plant growth and immunity. In the last few decades, a vast amount of information has been obtained detailing the role of different plant hormones in immunity, and how they work together to ultimately shape the outcomes of plant pathogen interactions. Here we provide an overview on the roles of the main classes of plant hormones in the regulation of plant immunity, highlighting their metabolic and signaling pathways and how plants and pathogens utilize these pathways to activate or suppress defence. Copyright © 2016 Elsevier Ltd. All rights reserved.

Inactivation of Mycobacterium bovis ssp. caprae in high-temperature, short-term pasteurized pilot-plant milk.

PubMed

Hammer, P; Richter, E; Rüsch-Gerdes, S; Walte, H-G C; Matzen, S; Kiesner, C

2015-03-01

Experiments to determine the efficacy of high temperature, short time (HTST) pasteurization of milk in terms of inactivation of pathogenic microorganisms were mainly performed between 1930 and 1960. Among the target organisms were Mycobacterium bovis and Mycobacterium tuberculosis. As a result, the Codex Alimentarius prescribes that HTST treatment of milk should lead to a significant reduction of pathogenic microorganisms during milk pasteurization. Due to the development of improved methods for the detection of survivors and of more advanced heating technology, verification of this requirement seemed to be necessary. To address recent outbreaks of tuberculosis in cattle caused by M. bovis ssp. caprae (M. caprae) in the southern regions of Germany, this organism was tested and compared with M. bovis ssp. bovis (M. bovis). Experiments were performed in a pilot plant for HTST pasteurization of milk with 3 strains of M. caprae and 1 strain of M. bovis. In preliminary trials at a fixed holding time of 25 s, the temperature at which significant inactivation occurred was 62.5°C for all strains. To determine D-values (decimal reduction times) for the inactivation kinetics, the strains were tested at 65, 62.5, and 60°C at holding times of 16.5, 25, and 35 s. At 65°C, the D-values of all strains ranged from 6.8 to 7.8 s, and at 62.5°C, D-values ranged from 14.5 to 18.1 s. Low inactivation was observed at 60°C. When the low slope of the inactivation curve allowed calculation of a D-value, these ranged from 40.8 to 129.9 s. In terms of log10 reductions, the highest values for all strains were 4.1 to 4.9 log at 65°C, with a holding time of 35 s. The tested strains of M. caprae and M. bovis showed similar low resistance to heat. Standard HTST treatment should result in a high reduction of these organisms and thus the requirements of the Codex Alimentarius for inactivation of pathogens by this process are far exceeded. Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Cronobacter sakazakii reduction by blueberry proanthocyanidins.

PubMed

Joshi, Snehal S; Howell, Amy B; D'Souza, Doris H

2014-05-01

Blueberry juice and blueberry polyphenols reportedly have antimicrobial properties against foodborne pathogens, without much currently known on their effects against Cronobacter sakazakii. This study evaluated the antimicrobial effects of blueberry proanthocyanidins (PAC) and commercial blueberry juice (BJ) against two strains of C. sakazakii, ATCC 29004 and 29544. BJ (pH 2.8), blueberry PAC (5 mg/ml) and controls (phosphate buffered saline (PBS), pH 7.2, and malic acid pH 3.0) were mixed with equal volumes of washed overnight cultures of C. sakazakii and incubated for 30 min, 1 h, 3 h and 6 h at 37°C. Reductions of ∼1 and 1.50 log CFU/ml were obtained for strains 29004 and 29544, respectively after 30 min with BJ or blueberry PAC. Both C. sakazakii strains 29004 and 29544 were reduced to undetectable levels from 8.25 ± 0.12 log CFU/ml and 8.48 ± 0.03 log CFU/ml, respectively with BJ (pH 2.8) or blueberry PAC after 1 h, while malic acid (pH 3.0) showed ∼1.3 log CFU/ml reduction for both strains. Scanning electron microscopy studies showed differences in cell membrane morphology with clumping and formation of blebs of the treated strains compared to untreated controls. These results warrant further in vivo studies with blueberry bioactives to determine potential for preventing and treating C. sakazakii infections. Copyright © 2013. Published by Elsevier Ltd.

Rapid, non-invasive imaging of alphaviral brain infection: reducing animal numbers and morbidity to identify efficacy of potential vaccines and antivirals.

PubMed

Patterson, Michael; Poussard, Allison; Taylor, Katherine; Seregin, Alexey; Smith, Jeanon; Peng, Bi-Hung; Walker, Aida; Linde, Jenna; Smith, Jennifer; Salazar, Milagros; Paessler, Slobodan

2011-11-21

Rapid and accurate identification of disease progression are key factors in testing novel vaccines and antivirals against encephalitic alphaviruses. Typical efficacy studies utilize a large number of animals and severe morbidity or mortality as an endpoint. New technologies provide a means to reduce and refine the animal use as proposed in Hume's 3Rs (replacement, reduction, refinement) described by Russel and Burch. In vivo imaging systems (IVIS) and bioluminescent enzyme technologies accomplish the reduction of animal requirements while shortening the experimental time and improving the accuracy in localizing active virus replication. In the case of murine models of viral encephalitis in which central nervous system (CNS) viral invasion occurs rapidly but the disease development is relatively slow, we visualized the initial brain infection and enhance the data collection process required for efficacy studies on antivirals or vaccines that are aimed at preventing brain infection. Accordingly, we infected mice through intranasal inoculation with the genetically modified pathogen, Venezuelan equine encephalitis, which expresses a luciferase gene. In this study, we were able to identify the invasion of the CNS at least 3 days before any clinical signs of disease, allowing for reduction of animal morbidity providing a humane means of disease and vaccine research while obtaining scientific data accurately and more rapidly. Based on our data from the imaging model, we confirmed the usefulness of this technology in preclinical research by demonstrating the efficacy of Ampligen, a TLR-3 agonist, in preventing CNS invasion. Copyright © 2011 Elsevier Ltd. All rights reserved.

Distribution and Genetic Variability of Fusarium oxysporum Associated with Tomato Diseases in Algeria and a Biocontrol Strategy with Indigenous Trichoderma spp.

PubMed

Debbi, Ali; Boureghda, Houda; Monte, Enrique; Hermosa, Rosa

2018-01-01

Fifty fungal isolates were sampled from diseased tomato plants as result of a survey conducted in seven tomato crop areas in Algeria from 2012 to 2015. Morphological criteria and PCR-based identification, using the primers PF02 and PF03, assigned 29 out of 50 isolates to Fusarium oxysporum ( Fo ). The banding patterns amplified for genes SIX1, SIX3 and SIX4 served to identify races 2 and 3 of Fo f. sp. lycopersici (FOL), and Fo f. sp. radicis lycopersici (FORL) among the Algerian isolates. All FOL isolates showed pathogenicity on the susceptible tomato cv. "Super Marmande," while nine of out 10 Algerian FORL isolates were pathogenic on tomato cv. "Rio Grande." Inter simple sequence repeat (ISSR) fingerprints showed high genetic diversity among Algerian Fo isolates. Seventeen Algerian Trichoderma isolates were also obtained and assigned to the species T. asperellum (12 isolates), T. harzianum (four isolates) and T. ghanense (one isolate) based on ITS and tef1 α gene sequences. Different in vitro tests identified the antagonistic potential of native Trichoderma isolates against FORL and FOL. Greenhouse biocontrol assays performed on "SM" tomato plants with T. ghanense T8 and T. asperellum T9 and T17, and three Fo isolates showed that isolate T8 performed well against FORL and FOL. This finding was based on an incidence reduction of crown and root rot and Fusarium wilt diseases by 53.1 and 48.3%, respectively.

Trehalose-6-Phosphate as a Potential Lead Candidate for the Development of Tps1 Inhibitors: Insights from the Trehalose Biosynthesis Pathway in Diverse Yeast Species.

PubMed

Magalhães, Rayne S S; De Lima, Karina C; de Almeida, Diego S G; De Mesquita, Joelma F; Eleutherio, Elis C A

2017-03-01

In some pathogens, trehalose biosynthesis is induced in response to stress as a protection mechanism. This pathway is an attractive target for antimicrobials as neither the enzymes, Tps1, and Tps2, nor is trehalose present in humans. Accumulation of T6P in Candida albicans, achieved by deletion of TPS2, resulted in strong reduction of fungal virulence. In this work, the effect of T6P on Tps1 activity was evaluated. Saccharomyces cerevisiae, C. albicans, and Candida tropicalis were used as experimental models. As expected, a heat stress induced both trehalose accumulation and increased Tps1 activity. However, the addition of 125 μM T6P to extracts obtained from stressed cells totally abolished or reduced in 50 and 60 % the induction of Tps1 activity in S. cerevisiae, C. tropicalis, and C. albicans, respectively. According to our results, T6P is an uncompetitive inhibitor of S. cerevisiae Tps1. This kind of inhibitor is able to decrease the rate of reaction to zero at increased concentrations. Based on the similarities found in sequence and function between Tps1 of S. cerevisiae and some pathogens and on the inhibitory effect of T6P on Tps1 activity observed in vitro, novel drugs can be developed for the treatment of infectious diseases caused by organisms whose infectivity and survival on the host depend on trehalose.

A Sensitive Assay for Virus Discovery in Respiratory Clinical Samples

PubMed Central

de Vries, Michel; Deijs, Martin; Canuti, Marta; van Schaik, Barbera D. C.; Faria, Nuno R.; van de Garde, Martijn D. B.; Jachimowski, Loes C. M.; Jebbink, Maarten F.; Jakobs, Marja; Luyf, Angela C. M.; Coenjaerts, Frank E. J.; Claas, Eric C. J.; Molenkamp, Richard; Koekkoek, Sylvie M.; Lammens, Christine; Leus, Frank; Goossens, Herman; Ieven, Margareta; Baas, Frank; van der Hoek, Lia

2011-01-01

In 5–40% of respiratory infections in children, the diagnostics remain negative, suggesting that the patients might be infected with a yet unknown pathogen. Virus discovery cDNA-AFLP (VIDISCA) is a virus discovery method based on recognition of restriction enzyme cleavage sites, ligation of adaptors and subsequent amplification by PCR. However, direct discovery of unknown pathogens in nasopharyngeal swabs is difficult due to the high concentration of ribosomal RNA (rRNA) that acts as competitor. In the current study we optimized VIDISCA by adjusting the reverse transcription enzymes and decreasing rRNA amplification in the reverse transcription, using hexamer oligonucleotides that do not anneal to rRNA. Residual cDNA synthesis on rRNA templates was further reduced with oligonucleotides that anneal to rRNA but can not be extended due to 3′-dideoxy-C6-modification. With these modifications >90% reduction of rRNA amplification was established. Further improvement of the VIDISCA sensitivity was obtained by high throughput sequencing (VIDISCA-454). Eighteen nasopharyngeal swabs were analysed, all containing known respiratory viruses. We could identify the proper virus in the majority of samples tested (11/18). The median load in the VIDISCA-454 positive samples was 7.2 E5 viral genome copies/ml (ranging from 1.4 E3–7.7 E6). Our results show that optimization of VIDISCA and subsequent high-throughput-sequencing enhances sensitivity drastically and provides the opportunity to perform virus discovery directly in patient material. PMID:21283679

Spatial dynamics of bar-headed geese migration in the context of H5N1

USGS Publications Warehouse

Bourouiba, L.; Wu, Jianhong; Newman, S.; Takekawa, John Y.; Natdorj, T.; Batbayar, N.; Bishop, C.M.; Hawkes, L.A.; Butler, P.J.; Wikelski, M.

2010-01-01

Virulent outbreaks of highly pathogenic avian influenza (HPAI) since 2005 have raised the question about the roles of migratory and wild birds in the transmission of HPAI. Despite increased monitoring, the role of wild waterfowl as the primary source of the highly pathogenic H5N1 has not been clearly established. The impact of outbreaks of HPAI among species of wild birds which are already endangered can nevertheless have devastating consequences for the local and non-local ecology where migratory species are established. Understanding the entangled dynamics of migration and the disease dynamics will be key to prevention and control measures for humans, migratory birds and poultry. Here, we present a spatial dynamic model of seasonal migration derived from first principles and linking the local dynamics during migratory stopovers to the larger scale migratory routes. We discuss the effect of repeated epizootic at specific migratory stopovers for bar-headed geese (Anser indicus). We find that repeated deadly outbreaks of H5N1 on stopovers during the autumn migration of bar-headed geese could lead to a larger reduction in the size of the equilibrium bird population compared with that obtained after repeated outbreaks during the spring migration. However, the opposite is true during the first few years of transition to such an equilibrium. The age-maturation process of juvenile birds which are more susceptible to H5N1 reinforces this result.

Disinfection of radish and alfalfa seeds inoculated with Escherichia coli O157:H7 and Salmonella by a gaseous acetic acid treatment.

PubMed

Nei, Daisuke; Latiful, Bari M; Enomoto, Katsuyoshi; Inatsu, Yasuhiro; Kawamoto, Shinnichi

2011-10-01

Abstract The majority of seed sprout-related outbreaks have been associated with Escherichia coli O157:H7 and Salmonella. Therefore, we aimed to find an effective method to inactivate these organisms on seeds before sprouting. Treatment with 8.7% (v/v) acetic acid at 55°C for 2-3 h reduced the population of E. coli O157:H7 and Salmonella inoculated on alfalfa (Medicago sativa L.) and radish seeds (Raphanus sativus L.) by more than 5.0 log CFU/g, and a longer treatment time completely eliminated the E. coli O157:H7 population. The E. coli O157:H7 populations were reduced to an undetectable level with a gaseous acetic acid treatment for 48 h. After enrichment, no E. coli O157:H7 were found in the alfalfa and radish seeds (25 g). However, these treatments were unable to eliminate Salmonella in both seed types. No significant difference between the germination rates of treated alfalfa seeds and control seeds was found, and germination rates greater than 95% were obtained for the radish seeds. Although chlorine washing is commonly used for seed decontamination, chlorine washing at 200 and 20,000 ppm resulted in a reduction of pathogens by less than or equal to 3 log CFU/g. Therefore, these results suggested that gaseous acetic acid is more effective than chlorine washing in controlling pathogenic bacteria on sprout seeds.

Inactivation kinetics of foodborne pathogens by UV-C radiation and its subsequent growth in fresh-cut kailan-hybrid broccoli.

PubMed

Martínez-Hernández, Ginés Benito; Huertas, Juan-Pablo; Navarro-Rico, Javier; Gómez, Perla A; Artés, Francisco; Palop, Alfredo; Artés-Hernández, Francisco

2015-04-01

The inactivation of Escherichia coli, S. Enteritidis and Listeria monocytogenes after UV-C radiation with 0, 2.5, 5, 7.5, 10 and 15 kJ UV-C m(-2) on fresh-cut kailan-hybrid broccoli was explored. Inactivation did not follow linear kinetics. Hence, it was modelled by using the Weibull distribution function, obtaining adjusted R(2) values higher than 94%, indicative of the accuracy of the model to the experimental data. The UV-C doses needed to reduce 1 log cycle the E. coli, S. Enteritidis and L. monocytogenes counts were 1.07, 0.02 and 9.26 kJ m(-2), respectively, being S. Enteritidis the most sensitive microorganism to UV-C radiation while L. monocytogenes was the most resistant. According to experimental data, UV-C doses higher than 2.5 kJ m(-2) did not achieve great microbial reductions. No differences in the growth behaviour of these microorganisms was observed in the treated samples stored under air conditions at 5, 10 and 15 °C, compared to the control. Conclusively, low UV-C doses are effective to reduce E. coli, S. Enteritidis and L. monocytogenes populations in fresh-cut kailan-hybrid broccoli keeping such counts stable during shelf life at 5-10 °C. The current study provides inactivation models for these foodborne pathogens that can be used in microbial risk assessment. Copyright © 2014 Elsevier Ltd. All rights reserved.

Integrated Evaluation Concept to Assess the Efficacy of Advanced Wastewater Treatment Processes for the Elimination of Micropollutants and Pathogens.

PubMed

Ternes, Thomas A; Prasse, Carsten; Eversloh, Christian Lütke; Knopp, Gregor; Cornel, Peter; Schulte-Oehlmann, Ulrike; Schwartz, Thomas; Alexander, Johannes; Seitz, Wolfram; Coors, Anja; Oehlmann, Jörg

2017-01-03

A multidisciplinary concept has been developed to compare advanced wastewater treatment processes for their efficacy of eliminating micropollutants and pathogens. The concept is based on (i) the removal/formation of selected indicator substances and their transformation products (TPs), (ii) the assessment of ecotoxicity via in vitro tests, and (iii) the removal of pathogens and antibiotic resistant bacteria. It includes substances passing biological wastewater treatment plants regulated or proposed to be regulated in the European Water Framework Directive, TPs formed in biological processes or during ozonation, agonistic/antagonistic endocrine activities, mutagenic/genotoxic activities, cytotoxic activities, further activities like neurotoxicity as well as antibiotics resistance genes, and taxonomic gene markers for pathogens. At a pilot plant, ozonation of conventionally treated wastewater resulted in the removal of micropollutants and pathogens and the reduction of estrogenic effects, whereas the in vitro mutagenicity increased. Subsequent post-treatment of the ozonated water by granular activated carbon (GAC) significantly reduced the mutagenic effects as well as the concentrations of remaining micropollutants, whereas this was not the case for biofiltration. The results demonstrate the suitability of the evaluation concept to assess processes of advanced wastewater treatment including ozonation and GAC by considering chemical, ecotoxicological, and microbiological parameters.

Development of a DNA Microarray-Based Assay for the Detection of Sugar Beet Root Rot Pathogens.

PubMed

Liebe, Sebastian; Christ, Daniela S; Ehricht, Ralf; Varrelmann, Mark

2016-01-01

Sugar beet root rot diseases that occur during the cropping season or in storage are accompanied by high yield losses and a severe reduction of processing quality. The vast diversity of microorganism species involved in rot development requires molecular tools allowing simultaneous identification of many different targets. Therefore, a new microarray technology (ArrayTube) was applied in this study to improve diagnosis of sugar beet root rot diseases. Based on three marker genes (internal transcribed spacer, translation elongation factor 1 alpha, and 16S ribosomal DNA), 42 well-performing probes enabled the identification of prevalent field pathogens (e.g., Aphanomyces cochlioides), storage pathogens (e.g., Botrytis cinerea), and ubiquitous spoilage fungi (e.g., Penicillium expansum). All probes were proven for specificity with pure cultures from 73 microorganism species as well as for in planta detection of their target species using inoculated sugar beet tissue. Microarray-based identification of root rot pathogens in diseased field beets was successfully confirmed by classical detection methods. The high discriminatory potential was proven by Fusarium species differentiation based on a single nucleotide polymorphism. The results demonstrate that the ArrayTube constitute an innovative tool allowing a rapid and reliable detection of plant pathogens particularly when multiple microorganism species are present.

Pathogenic amoebae in power-plant cooling lakes. Final report

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tyndall, R.L.; Willaert, E.; Stevens, A.R.

1981-06-01

Cooling waters and associated algae and sediments from four northern and four southern/western electric power plants were tested for the presence of pathogenic amoebae. Unheated control waters and algae/sediments from four northern and five southern/western sites were also tested. When comparing results from the test versus control sites, a significantly higher proportion (P less than or equal to 0.05) of the samples from the test sites were positive for thermophilic amoeba, thermophilic Naegleria and pathogenic Naegleria. The difference in number of samples positive for thermophilic Naegleria between heated and unheated waters, however, was attributable predominantly to the northern waters andmore » algae/sediments. While two of four northern test sites yielded pathogenic Naegleria, seven of the eight isolates were obtained from one site. Seasonality effects relative to the isolation of the pathogen were also noted at this site. One pathogen was isolated from a southwestern test site. Pathogens were not isolated from any control sites. Some of the pathogenic isolates were analyzed serologically and classified as pathogenic Naegleria fowleri. Salinity, pH, conductivity, and bacteriological profiles did not obviously correlate with the presence or absence of pathogenic Naegleria. While thermal addition was significantly associated with the presence of thermophilic Naegleria (P less than or equal to 0.05), the data implicate other as yet undefined parameters associated with the presence of the pathogenic thermophile. Until further delineation of these parameters is effected, generalizations cannot be made concerning the effect of thermal impact on the growth of pathogenic amoeba in a particular cooling system.« less

A Role for a Menthone Reductase in Resistance against Microbial Pathogens in Plants1[C][W][OA

PubMed Central

Choi, Hyong Woo; Lee, Byung Gil; Kim, Nak Hyun; Park, Yong; Lim, Chae Woo; Song, Hyun Kyu; Hwang, Byung Kook

2008-01-01

Plants elaborate a vast array of enzymes that synthesize defensive secondary metabolites in response to pathogen attack. Here, we isolated the pathogen-responsive CaMNR1 [menthone: (+)-(3S)-neomenthol reductase] gene, a member of the short-chain dehydrogenase/reductase (SDR) superfamily, from pepper (Capsicum annuum) plants. Gas chromatography-mass spectrometry analysis revealed that purified CaMNR1 and its ortholog AtSDR1 from Arabidopsis (Arabidopsis thaliana) catalyze a menthone reduction with reduced nicotinamide adenine dinucleotide phosphate as a cofactor to produce neomenthol with antimicrobial activity. CaMNR1 and AtSDR1 also possess a significant catalytic activity for neomenthol oxidation. We examined the cellular function of the CaMNR1 gene by virus-induced gene silencing and ectopic overexpression in pepper and Arabidopsis plants, respectively. CaMNR1-silenced pepper plants were significantly more susceptible to Xanthomonas campestris pv vesicatoria and Colletotrichum coccodes infection and expressed lower levels of salicylic acid-responsive CaBPR1 and CaPR10 and jasmonic acid-responsive CaDEF1. CaMNR1-overexpressing Arabidopsis plants exhibited enhanced resistance to the hemibiotrophic pathogen Pseudomonas syringae pv tomato DC3000 and the biotrophic pathogen Hyaloperonospora parasitica isolate Noco2, accompanied by the induction of AtPR1 and AtPDF1.2. In contrast, mutation in the CaMNR1 ortholog AtSDR1 significantly enhanced susceptibility to both pathogens. Together, these results indicate that the novel menthone reductase gene CaMNR1 and its ortholog AtSDR1 positively regulate plant defenses against a broad spectrum of pathogens. PMID:18599651

Individual and co-operative roles of lactic acid and hydrogen peroxide in the killing activity of enteric strain Lactobacillus johnsonii NCC933 and vaginal strain Lactobacillus gasseri KS120.1 against enteric, uropathogenic and vaginosis-associated pathogens.

PubMed

Atassi, Fabrice; Servin, Alain L

2010-03-01

The mechanism underlying the killing activity of Lactobacillus strains against bacterial pathogens appears to be multifactorial. Here, we investigate the respective contributions of hydrogen peroxide and lactic acid in killing bacterial pathogens associated with the human vagina, urinary tract or intestine by two hydrogen peroxide-producing strains. In co-culture, the human intestinal strain Lactobacillus johnsonii NCC933 and human vaginal strain Lactobacillus gasseri KS120.1 strains killed enteric Salmonella enterica serovar Typhimurium SL1344, vaginal Gardnerella vaginalis DSM 4944 and urinary tract Escherichia coli CFT073 pathogens. The cell-free culture supernatants (CFCSs) produced the same reduction in SL1344, DSM 4944 and CFT073 viability, whereas isolated bacteria had no effect. The killing activity of CFCSs was heat-stable. In the presence of Dulbecco's modified Eagle's minimum essential medium inhibiting the lactic acid-dependent killing activity, CFCSs were less effective at killing of the pathogens. Catalase-treated CFCSs displayed a strong decreased activity. Tested alone, hydrogen peroxide triggered a concentration-dependent killing activity against all three pathogens. Lactic acid alone developed a killing activity only at concentrations higher than that present in CFCSs. In the presence of lactic acid at a concentration present in Lactobacillus CFCSs, hydrogen peroxide displayed enhanced killing activity. Collectively, these results demonstrate that for hydrogen peroxide-producing Lactobacillus strains, the main metabolites of Lactobacillus, lactic acid and hydrogen peroxide, act co-operatively to kill enteric, vaginosis-associated and uropathogenic pathogens.

Hand hygiene regimens for the reduction of risk in food service environments.

PubMed

Edmonds, Sarah L; McCormack, Robert R; Zhou, Sifang Steve; Macinga, David R; Fricker, Christopher M

2012-07-01

Pathogenic strains of Escherichia coli and human norovirus are the main etiologic agents of foodborne illness resulting from inadequate hand hygiene practices by food service workers. This study was conducted to evaluate the antibacterial and antiviral efficacy of various hand hygiene product regimens under different soil conditions representative of those in food service settings and assess the impact of product formulation on this efficacy. On hands contaminated with chicken broth containing E. coli, representing a moderate soil load, a regimen combining an antimicrobial hand washing product with a 70% ethanol advanced formula (EtOH AF) gel achieved a 5.22-log reduction, whereas a nonantimicrobial hand washing product alone achieved a 3.10log reduction. When hands were heavily soiled from handling ground beef containing E. coli, a wash-sanitize regimen with a 0.5% chloroxylenol antimicrobial hand washing product and the 70% EtOH AF gel achieved a 4.60-log reduction, whereas a wash-sanitize regimen with a 62% EtOH foam achieved a 4.11-log reduction. Sanitizing with the 70% EtOH AF gel alone was more effective than hand washing with a nonantimicrobial product for reducing murine norovirus (MNV), a surrogate for human norovirus, with 2.60- and 1.79-log reductions, respectively. When combined with hand washing, the 70% EtOH AF gel produced a 3.19-log reduction against MNV. A regimen using the SaniTwice protocol with the 70% EtOH AF gel produced a 4.04-log reduction against MNV. These data suggest that although the process of hand washing helped to remove pathogens from the hands, use of a wash-sanitize regimen was even more effective for reducing organisms. Use of a high-efficacy sanitizer as part of a wash-sanitize regimen further increased the efficacy of the regimen. The use of a well-formulated alcohol-based hand rub as part of a wash-sanitize regimen should be considered as a means to reduce risk of infection transmission in food service facilities.

Evaluation of advanced wastewater treatment systems for water reuse in the era of advanced wastewater treatment

NASA Astrophysics Data System (ADS)

Kon, Hisao; Watanabe, Masahiro

This study focuses on effluent COD concentration from wastewater treatment in regards to the reduction of pathogenic bacteria and trace substances in public waters. The main types of secondary wastewater treatment were conventional activated sludge processes. Recently, however, advance wastewater treatment processes have been developed aimed at the removal of nitrogen and phosphorus, and the effluent quality of these processes was analyzed in this study. Treatment processes for water reclamation that make effluent to meet the target water quality for reuse purposes were selected and also optimum design parameters for these processes were proposed. It was found that the treatment cost to water reclamation was greatly affected by the effluent COD of the secondary treatment. It is important to maintain low COD concentration in the secondary treated effluent. Therefore, it is considered that adequate cost benefits would be obtained by achieving target COD quality through shifting from a conventional activated sludge process to an advanced treatment process.

Green Route for Silver Nanoparticles Synthesis by Raphanus Sativus Extract in a Continuous Flow Tubular Microreactor

NASA Astrophysics Data System (ADS)

Jolhe, P. D.; Bhanvase, B. A.; Patil, V. S.; Sonawane, S. H.

The present work deals with the investigation of the greener route for the production of silver nanoparticles using Raphanus sativus (R. sativus) bioextract in a continuous flow tubular microreactor. The parameters affecting the particle size and distribution were investigated. From the results obtained it can be inferred that the ascorbic acid (reducing agent) present in the R. sativus bioextract is responsible for the reduction of silver ions. At optimum condition, the particle size distribution of nanoparticles is found between 18nm and 39nm. The absorbance value was found to be decreased with an increase in the diameter of the microreactor. It indicates that a number of nuclei are formed in the micrometer sized (diameter) reactor because of the better solute transfer rate leading to the formation of large number of silver nanoparticles. The study of antibacterial activity of green synthesized silver nanoparticles shows effective inhibitory activity against waterborne pathogens, Shegella and Listeria bacteria.

Nosema ceranae in South American Native Stingless Bees and Social Wasp.

PubMed

Porrini, Martín Pablo; Porrini, Leonardo Pablo; Garrido, Paula Melisa; de Melo E Silva Neto, Carlos; Porrini, Darío Pablo; Muller, Fernando; Nuñez, Laura Alejandra; Alvarez, Leopoldo; Iriarte, Pedro Fernandez; Eguaras, Martín Javier

2017-11-01

Besides the incipient research effort, the role of parasites as drivers of the reduction affecting pollinator populations is mostly unknown. Given the worldwide extension of the beekeeping practice and the diversity of pathogens affecting Apis mellifera populations, honey bee colonies are a certain source of parasite dispersion to other species. Here, we communicate the detection of the microsporidium Nosema ceranae, a relatively new parasite of honey bees, in stingless bees (Meliponini) and the social wasp Polybia scutellaris (Vespidae) samples from Argentina and Brazil by means of duplex PCR. Beyond the geographic location of the nests, N. ceranae was detected in seven from the eight Meliponini species analyzed, while Nosema apis, another common parasite of A. mellifera, was absent in all samples tested. Further research is necessary to determine if the presence of the parasite is also associated with established infection in host tissues. The obtained information enriches the current knowledge about pathologies that can infect or, at least, be vectored by native wild pollinators from South America.

Deep-sea vent chemoautotrophs: diversity, biochemistry and ecological significance.

PubMed

Nakagawa, Satoshi; Takai, Ken

2008-07-01

Deep-sea vents support productive ecosystems driven primarily by chemoautotrophs. Chemoautotrophs are organisms that are able to fix inorganic carbon using a chemical energy obtained through the oxidation of reduced compounds. Following the discovery of deep-sea vent ecosystems in 1977, there has been an increasing knowledge that deep-sea vent chemoautotrophs display remarkable physiological and phylogenetic diversity. Cultivation-dependent and -independent studies have led to an emerging view that the majority of deep-sea vent chemoautotrophs have the ability to derive energy from a variety of redox couples other than the conventional sulfur-oxygen couple, and fix inorganic carbon via the reductive tricarboxylic acid cycle. In addition, recent genomic, metagenomic and postgenomic studies have considerably accelerated the comprehensive understanding of molecular mechanisms of deep-sea vent chemoautotrophy, even in yet uncultivable endosymbionts of vent fauna. Genomic analysis also suggested that there are previously unrecognized evolutionary links between deep-sea vent chemoautotrophs and important human/animal pathogens. This review summarizes chemoautotrophy in deep-sea vents, highlighting recent biochemical and genomic discoveries.

Westward Spread of Highly Pathogenic Avian Influenza A(H7N9) Virus among Humans, China.

PubMed

Yang, Qiqi; Shi, Wei; Zhang, Lei; Xu, Yi; Xu, Jing; Li, Shen; Zhang, Junjun; Hu, Kan; Ma, Chaofeng; Zhao, Xiang; Li, Xiyan; Liu, Feng; Tong, Xin; Zhang, Guogang; Yu, Pengbo; Pybus, Oliver G; Tian, Huaiyu

2018-06-01

We report infection of humans with highly pathogenic avian influenza A(H7N9) virus in Shaanxi, China, in May 2017. We obtained complete genomes for samples from 5 patients and from live poultry markets or farms in 4 cities. Results indicate that H7N9 is spreading westward from southern and eastern China.

The compact genome of the plant pathogen Plasmodiophora brassicae is adapted to intracellular interactions with host Brassica spp.

PubMed

Rolfe, Stephen A; Strelkov, Stephen E; Links, Matthew G; Clarke, Wayne E; Robinson, Stephen J; Djavaheri, Mohammad; Malinowski, Robert; Haddadi, Parham; Kagale, Sateesh; Parkin, Isobel A P; Taheri, Ali; Borhan, M Hossein

2016-03-31

The protist Plasmodiophora brassicae is a soil-borne pathogen of cruciferous species and the causal agent of clubroot disease of Brassicas including agriculturally important crops such as canola/rapeseed (Brassica napus). P. brassicae has remained an enigmatic plant pathogen and is a rare example of an obligate biotroph that resides entirely inside the host plant cell. The pathogen is the cause of severe yield losses and can render infested fields unsuitable for Brassica crop growth due to the persistence of resting spores in the soil for up to 20 years. To provide insight into the biology of the pathogen and its interaction with its primary host B. napus, we produced a draft genome of P. brassicae pathotypes 3 and 6 (Pb3 and Pb6) that differ in their host range. Pb3 is highly virulent on B. napus (but also infects other Brassica species) while Pb6 infects only vegetable Brassica crops. Both the Pb3 and Pb6 genomes are highly compact, each with a total size of 24.2 Mb, and contain less than 2 % repetitive DNA. Clustering of genome-wide single nucleotide polymorphisms (SNP) of Pb3, Pb6 and three additional re-sequenced pathotypes (Pb2, Pb5 and Pb8) shows a high degree of correlation of cluster grouping with host range. The Pb3 genome features significant reduction of intergenic space with multiple examples of overlapping untranslated regions (UTRs). Dependency on the host for essential nutrients is evident from the loss of genes for the biosynthesis of thiamine and some amino acids and the presence of a wide range of transport proteins, including some unique to P. brassicae. The annotated genes of Pb3 include those with a potential role in the regulation of the plant growth hormones cytokinin and auxin. The expression profile of Pb3 genes, including putative effectors, during infection and their potential role in manipulation of host defence is discussed. The P. brassicae genome sequence reveals a compact genome, a dependency of the pathogen on its host for some essential nutrients and a potential role in the regulation of host plant cytokinin and auxin. Genome annotation supported by RNA sequencing reveals significant reduction in intergenic space which, in addition to low repeat content, has likely contributed to the P. brassicae compact genome.

Concurrent host-pathogen gene expression in the lungs of pigs challenged with Actinobacillus pleuropneumoniae.

PubMed

Brogaard, Louise; Klitgaard, Kirstine; Heegaard, Peter M H; Hansen, Mette Sif; Jensen, Tim Kåre; Skovgaard, Kerstin

2015-05-28

Actinobacillus pleuropneumoniae causes pleuropneumonia in pigs, a disease which is associated with high morbidity and mortality, as well as impaired animal welfare. To obtain in-depth understanding of this infection, the interplay between virulence factors of the pathogen and defense mechanisms of the porcine host needs to be elucidated. However, research has traditionally focused on either bacteriology or immunology; an unbiased picture of the transcriptional responses can be obtained by investigating both organisms in the same biological sample. Host and pathogen responses in pigs experimentally infected with A. pleuropneumoniae were analyzed by high-throughput RT-qPCR. This approach allowed concurrent analysis of selected genes encoding proteins known or hypothesized to be important in the acute phase of this infection. The expression of 17 bacterial and 31 porcine genes was quantified in lung samples obtained within the first 48 hours of infection. This provided novel insight into the early time course of bacterial genes involved in synthesis of pathogen-associated molecular patterns (lipopolysaccharide, peptidoglycan, lipoprotein) and genes involved in pattern recognition (TLR4, CD14, MD2, LBP, MYD88) in response to A. pleuropneumoniae. Significant up-regulation of proinflammatory cytokines such as IL1B, IL6, and IL8 was observed, correlating with protein levels, infection status and histopathological findings. Host genes encoding proteins involved in iron metabolism, as well as bacterial genes encoding exotoxins, proteins involved in adhesion, and iron acquisition were found to be differentially expressed according to disease progression. By applying laser capture microdissection, porcine expression of selected genes could be confirmed in the immediate surroundings of the invading pathogen. Microbial pathogenesis is the product of interactions between host and pathogen. Our results demonstrate the applicability of high-throughput RT-qPCR for the elucidation of dual-organism gene expression analysis during infection. We showed differential expression of 12 bacterial and 24 porcine genes during infection and significant correlation of porcine and bacterial gene expression. This is the first study investigating the concurrent transcriptional response of both bacteria and host at the site of infection during porcine respiratory infection.

Reduced Set of Virulence Genes Allows High Accuracy Prediction of Bacterial Pathogenicity in Humans

PubMed Central

Iraola, Gregorio; Vazquez, Gustavo; Spangenberg, Lucía; Naya, Hugo

2012-01-01

Although there have been great advances in understanding bacterial pathogenesis, there is still a lack of integrative information about what makes a bacterium a human pathogen. The advent of high-throughput sequencing technologies has dramatically increased the amount of completed bacterial genomes, for both known human pathogenic and non-pathogenic strains; this information is now available to investigate genetic features that determine pathogenic phenotypes in bacteria. In this work we determined presence/absence patterns of different virulence-related genes among more than finished bacterial genomes from both human pathogenic and non-pathogenic strains, belonging to different taxonomic groups (i.e: Actinobacteria, Gammaproteobacteria, Firmicutes, etc.). An accuracy of 95% using a cross-fold validation scheme with in-fold feature selection is obtained when classifying human pathogens and non-pathogens. A reduced subset of highly informative genes () is presented and applied to an external validation set. The statistical model was implemented in the BacFier v1.0 software (freely available at ), that displays not only the prediction (pathogen/non-pathogen) and an associated probability for pathogenicity, but also the presence/absence vector for the analyzed genes, so it is possible to decipher the subset of virulence genes responsible for the classification on the analyzed genome. Furthermore, we discuss the biological relevance for bacterial pathogenesis of the core set of genes, corresponding to eight functional categories, all with evident and documented association with the phenotypes of interest. Also, we analyze which functional categories of virulence genes were more distinctive for pathogenicity in each taxonomic group, which seems to be a completely new kind of information and could lead to important evolutionary conclusions. PMID:22916122

Comparative studies on single-layer reduced graphene oxide films obtained by electrochemical reduction and hydrazine vapor reduction

NASA Astrophysics Data System (ADS)

Wang, Zhijuan; Wu, Shixin; Zhang, Juan; Chen, Peng; Yang, Guocheng; Zhou, Xiaozhu; Zhang, Qichun; Yan, Qingyu; Zhang, Hua

2012-02-01

The comparison between two kinds of single-layer reduced graphene oxide (rGO) sheets, obtained by reduction of graphene oxide (GO) with the electrochemical method and hydrazine vapor reduction, referred to as E-rGO and C-rGO, respectively, is systematically studied. Although there is no morphology difference between the E-rGO and C-rGO films adsorbed on solid substrates observed by AFM, the reduction process to obtain the E-rGO and C-rGO films is quite different. In the hydrazine vapor reduction, the nitrogen element is incorporated into the obtained C-rGO film, while no additional element is introduced to the E-rGO film during the electrochemical reduction. Moreover, Raman spectra show that the electrochemical method is more effective than the hydrazine vapor reduction method to reduce the GO films. In addition, E-rGO shows better electrocatalysis towards dopamine than does C-rGO. This study is helpful for researchers to understand these two different reduction methods and choose a suitable one to reduce GO based on their experimental requirements.

Effective Antimicrobial Regimens for Use in Humans for Therapy of Bacillus anthracis Infections and Postexposure Prophylaxis†

PubMed Central

Deziel, Mark R.; Heine, Henry; Louie, Arnold; Kao, Mark; Byrne, William R.; Basset, Jennifer; Miller, Lynda; Bush, Karen; Kelly, Michael; Drusano, G. L.

2005-01-01

Expanded options for treatments directed against pathogens that can be used for bioterrorism are urgently needed. Treatment regimens directed against such pathogens can be identified only by using data derived from in vitro and animal studies. It is crucial that these studies reliably predict the efficacy of proposed treatments in humans. The objective of this study was to identify a levofloxacin treatment regimen that will serve as an effective therapy for Bacillus anthracis infections and postexposure prophylaxis. An in vitro hollow-fiber infection model that replicates the pharmacokinetic profile of levofloxacin observed in humans (half-life [t1/2], 7.5 h) or in animals, such as the mouse or the rhesus monkey (t1/2, ∼2 h), was used to evaluate a proposed indication for levofloxacin (500 mg once daily) for the treatment of Bacillus anthracis infections. The results obtained with the in vitro model served as the basis for the doses and the dose schedules that were evaluated in the mouse inhalational anthrax model. The effects of levofloxacin and ciprofloxacin treatment were compared to those of no treatment (untreated controls). The main outcome measure in the in vitro hollow-fiber infection model was a persistent reduction of culture density (≥4 log10 reduction) and prevention of the emergence of levofloxacin-resistant organisms. In the mouse inhalational anthrax model the main outcome measure was survival. The results indicated that levofloxacin given once daily with simulated human pharmacokinetics effectively sterilized Bacillus anthracis cultures. By using a simulated animal pharmacokinetic profile, a once-daily dosing regimen that provided a human-equivalent exposure failed to sterilize the cultures. Dosing regimens that “partially humanized” levofloxacin exposures within the constraints of animal pharmacokinetics reproduced the antimicrobial efficacy seen with human pharmacokinetics. In a mouse inhalational anthrax model, once-daily dosing was significantly inferior (survival end point) to regimens of dosing every 12 h or every 6 h with identical total daily levofloxacin doses. These results demonstrate the predictive value of the in vitro hollow-fiber infection model with respect to the success or the failure of treatment regimens in animals. Furthermore, the model permits the evaluation of treatment regimens that “humanize” antibiotic exposures in animal models, enhancing the confidence with which animal models may be used to reliably predict the efficacies of proposed antibiotic treatments in humans in situations (e.g., the release of pathogens as agents of bioterrorism or emerging infectious diseases) where human trials cannot be performed. A treatment regimen effective in rhesus monkeys was identified. PMID:16304178

The Pathogen-Host Interactions database (PHI-base): additions and future developments

PubMed Central

Urban, Martin; Pant, Rashmi; Raghunath, Arathi; Irvine, Alistair G.; Pedro, Helder; Hammond-Kosack, Kim E.

2015-01-01

Rapidly evolving pathogens cause a diverse array of diseases and epidemics that threaten crop yield, food security as well as human, animal and ecosystem health. To combat infection greater comparative knowledge is required on the pathogenic process in multiple species. The Pathogen-Host Interactions database (PHI-base) catalogues experimentally verified pathogenicity, virulence and effector genes from bacterial, fungal and protist pathogens. Mutant phenotypes are associated with gene information. The included pathogens infect a wide range of hosts including humans, animals, plants, insects, fish and other fungi. The current version, PHI-base 3.6, available at http://www.phi-base.org, stores information on 2875 genes, 4102 interactions, 110 host species, 160 pathogenic species (103 plant, 3 fungal and 54 animal infecting species) and 181 diseases drawn from 1243 references. Phenotypic and gene function information has been obtained by manual curation of the peer-reviewed literature. A controlled vocabulary consisting of nine high-level phenotype terms permits comparisons and data analysis across the taxonomic space. PHI-base phenotypes were mapped via their associated gene information to reference genomes available in Ensembl Genomes. Virulence genes and hotspots can be visualized directly in genome browsers. Future plans for PHI-base include development of tools facilitating community-led curation and inclusion of the corresponding host target(s). PMID:25414340

Neelaredoxin, an iron-binding protein from the syphilis spirochete, Treponema pallidum, is a superoxide reductase.

PubMed

Jovanović, T; Ascenso, C; Hazlett, K R; Sikkink, R; Krebs, C; Litwiller, R; Benson, L M; Moura, I; Moura, J J; Radolf, J D; Huynh, B H; Naylor, S; Rusnak, F

2000-09-15

Treponema pallidum, the causative agent of venereal syphilis, is a microaerophilic obligate pathogen of humans. As it disseminates hematogenously and invades a wide range of tissues, T. pallidum presumably must tolerate substantial oxidative stress. Analysis of the T. pallidum genome indicates that the syphilis spirochete lacks most of the iron-binding proteins present in many other bacterial pathogens, including the oxidative defense enzymes superoxide dismutase, catalase, and peroxidase, but does possess an orthologue (TP0823) for neelaredoxin, an enzyme of hyperthermophilic and sulfate-reducing anaerobes shown to possess superoxide reductase activity. To analyze the potential role of neelaredoxin in treponemal oxidative defense, we examined the biochemical, spectroscopic, and antioxidant properties of recombinant T. pallidum neelaredoxin. Neelaredoxin was shown to be expressed in T. pallidum by reverse transcriptase-polymerase chain reaction and Western blot analysis. Recombinant neelaredoxin is a 26-kDa alpha(2) homodimer containing, on average, 0.7 iron atoms/subunit. Mössbauer and EPR analysis of the purified protein indicates that the iron atom exists as a mononuclear center in a mixture of high spin ferrous and ferric oxidation states. The fully oxidized form, obtained by the addition of K(3)(Fe(CN)(6)), exhibits an optical spectrum with absorbances at 280, 320, and 656 nm; the last feature is responsible for the protein's blue color, which disappears upon ascorbate reduction. The fully oxidized protein has a A(280)/A(656) ratio of 10.3. Enzymatic studies revealed that T. pallidum neelaredoxin is able to catalyze a redox equilibrium between superoxide and hydrogen peroxide, a result consistent with it being a superoxide reductase. This finding, the first description of a T. pallidum iron-binding protein, indicates that the syphilis spirochete copes with oxidative stress via a primitive mechanism, which, thus far, has not been described in pathogenic bacteria.

Thermal inactivation of Escherichia coli O157:H7 in blade-tenderized beef steaks cooked on a commercial open-flame gas grill.

PubMed

Luchansky, John B; Porto-Fett, Anna C S; Shoyer, Bradley; Phebus, Randall K; Thippareddi, Harshavardhan; Call, Jeffrey E

2009-07-01

Beef subprimals were inoculated on the lean side with ca. 4.0 log CFU/g of a cocktail of rifampin-resistant (Rif(r)) Escherichia coli O157:H7 strains and then passed once through a mechanical blade tenderizer with the lean side facing upward. Inoculated subprimals that were not tenderized served as controls. Two core samples were removed from each of three tenderized subprimals and cut into six consecutive segments starting from the inoculated side. A total of six cores were also obtained from control subprimals, but only segment 1 (topmost) was sampled. Levels of E. coli O157:H7 recovered from segment 1 were 3.81 log CFU/g for the control subprimals and 3.36 log CFU/g for tenderized subprimals. The percentage of cells recovered in segment 2 was ca. 25-fold lower than levels recovered from segment 1, but E. coli O157:H7 was recovered from all six segments of the cores obtained from tenderized subprimals. In phase II, lean-side-inoculated (ca. 4.0 log CFU/g), single-pass tenderized subprimals were cut into steaks of various thicknesses (1.91 cm [0.75 in.], 2.54 cm [1.0 in.], and 3.18 cm [1.25 in.]) that were subsequently cooked on a commercial open-flame gas grill to internal temperatures of 48.8 degrees C (120 degrees F), 54.4 degrees C (130 degrees F), and 60 degrees C (140 degrees F). In general, regardless of temperature or thickness, we observed about a 2.6- to 4.2-log CFU/g reduction in pathogen levels following cooking. These data validate that cooking on a commercial gas grill is effective at eliminating relatively low levels of the pathogen that may be distributed throughout a blade-tenderized steak.

Stepwise flow diagram for the development of formulations of non spore-forming bacteria against foliar pathogens: The case of Lysobacter capsici AZ78.

PubMed

Segarra, Guillem; Puopolo, Gerardo; Giovannini, Oscar; Pertot, Ilaria

2015-12-20

The formulation is a significant step in biopesticide development and is an efficient way to obtain consistency in terms of biological control under field conditions. Nonetheless, there is still a lack of information regarding the processes needed to achieve efficient formulation of non spore-forming bacterial biological control agents. In response to this, we propose a flow diagram made up of six steps including selection of growth parameters, checking of minimum shelf life, selection of protective additives, checking that the additives have no adverse effects, validation of the additive mix under field conditions and choosing whether to use additives as co-formulants or tank mix additives. This diagram is intended to provide guidance and decision-making criteria for the formulation of non spore-forming bacterial biological control agents against foliar pathogens. The diagram was then validated by designing an efficient formulation for a Gram-negative bacterium, Lysobacter capsici AZ78, to control grapevine downy mildew caused by Plasmopara viticola. A harvest of 10(10)L. capsici AZ78cellsml(-1) was obtained in a bench top fermenter. The viability of cells decreased by only one order of magnitude after one year of storage at 4°C. The use of a combination of corn steep liquor, lignosulfonate, and polyethyleneglycol in the formulation improved the survival of L. capsici AZ78 cells living on grapevine leaves under field conditions by one order of magnitude. Furthermore, the use of these additives also guaranteed a reduction of 71% in P. viticola attacks. In conclusion, this work presents a straightforward stepwise flow diagram to help researchers develop formulations for biological control agents that are easy to prepare, stable, not phytotoxic and able to protect the microorganims under field conditions. Copyright © 2015 Elsevier B.V. All rights reserved.

DAF-16-dependent suppression of immunity during reproduction in Caenorhabditis elegans.

PubMed

Miyata, Sachiko; Begun, Jakob; Troemel, Emily R; Ausubel, Frederick M

2008-02-01

To further understand how the nematode Caenorhabditis elegans defends itself against pathogen attack, we analyzed enhanced pathogen resistance (epr) mutants obtained from a forward genetic screen. We also examined several well-characterized sterile mutants that exhibit an Epr phenotype. We found that sterility and pathogen resistance are highly correlated and that resistance in both epr and sterile mutants is dependent on DAF-16 activity. Our data indicate that a DAF-16-dependent signaling pathway distinct from previously described pathways is involved in the activation of genes that confer resistance to bacterial pathogens. The timing of DAF-16-dependent gene activation in sterile mutants coincides with the onset of embryonic development in wild-type animals, suggesting that signals from developing embryos normally downregulate the immune response.

From Exit to Entry: Long-term Survival and Transmission of Salmonella

PubMed Central

Waldner, Landon L.; MacKenzie, Keith D.; Köster,, Wolfgang; White, Aaron P.

2012-01-01

Salmonella spp. are a leading cause of human infectious disease worldwide and pose a serious health concern. While we have an improving understanding of pathogenesis and the host-pathogen interactions underlying the infection process, comparatively little is known about the survival of pathogenic Salmonella outside their hosts. This review focuses on three areas: (1) in vitro evidence that Salmonella spp. can survive for long periods of time under harsh conditions; (2) observations and conclusions about Salmonella persistence obtained from human outbreaks; and (3) new information revealed by genomic- and population-based studies of Salmonella and related enteric pathogens. We highlight the mechanisms of Salmonella persistence and transmission as an essential part of their lifecycle and a prerequisite for their evolutionary success as human pathogens. PMID:25436767