In-toilet disinfection of fresh fecal sludge with ammonia naturally present in excreta

2015 ◽  
Vol 6 (1) ◽  
pp. 104-114 ◽  
Author(s):  
Temitope A. Ogunyoku ◽  
Fikreselam Habebo ◽  
Kara L. Nelson

A simple treatment method, Safe Sludge disinfection, was developed to disinfect pathogens in fresh fecal sludge using the ammonia naturally present in excreta. In the first step, urea is hydrolyzed to ammonia (NH3/NH4+). In the second step, Ca(OH)2 is added to raise the pH level such that NH3, a known disinfectant, is the dominant form of ammonia; subsequently, the waste is stored until sufficient disinfection is achieved. In a closed system at 23 °C, Safe Sludge disinfection achieved >9.3 log10 and >4.0 log10 decrease of indigenous Escherichia coli and seeded MS2 coliphage, respectively, within 10.6 hours, and 2.0 log10 inactivation of seeded Ascaris suum eggs within 2 weeks. Disinfection of feces at high pH with no urine addition was tested for comparison, and similar inactivation levels were achieved for E. coli and MS2 bacteriophage. However, for Ascaris eggs only 0.38 log10 inactivation was achieved over 2 weeks. For control samples (feces plus urine only), no inactivation of bacteria or virus indicators was observed and inactivation of Ascaris eggs was also low (0.42 log10). To illustrate how the Safe Sludge concept could be incorporated into a waterless household toilet, a conceptual design and prototype was developed, called the pHree Loo.

2005 ◽  
Vol 68 (9) ◽  
pp. 1895-1899 ◽  
Author(s):  
MANOJ KUMAR MOHAN NAIR ◽  
HANEM ABOUELEZZ ◽  
THOMAS HOAGLAND ◽  
KUMAR VENKITANARAYANAN

The antibacterial effect of low concentrations of monocaprylin on Escherichia coli O157:H7 in apple juice was investigated. Apple juice alone (control) or containing 2.5 mM (0.055%) or 5 mM monocaprylin was inoculated with a five-strain mixture of E. coli O157:H7 at ~6.0 log CFU/ml. The juice samples were stored at 23 or 4°C for 14 or 21 days, respectively, and the population of E. coli O157:H7 was determined on tryptic soy agar plates supplemented with 0.6% yeast extract. At both storage temperatures, the population of E. coli O157:H7 in monocaprylin-supplemented juice samples was significantly lower (P < 0.05) than that in the control samples. The concentration of monocaprylin and the storage temperature had a significant effect on the inactivation of E. coli O157:H7 in apple juice. Monocaprylin at 5 mM was significantly more effective than 2.5 mM monocaprylin for killing E. coli O157:H7 in apple juice. Inactivation of E. coli O157:H7 by monocaprylin was more pronounced in juice stored at 23°C than in the refrigerated samples. Results of this study indicated that monocaprylin is effective for killing E. coli O157:H7 in apple juice, but detailed sensory studies are needed to determine the organoleptic properties of apple juice containing monocaprylin.


2008 ◽  
Vol 74 (20) ◽  
pp. 6230-6238 ◽  
Author(s):  
Tamar Abuladze ◽  
Manrong Li ◽  
Marc Y. Menetrez ◽  
Timothy Dean ◽  
Andre Senecal ◽  
...  

ABSTRACT A bacteriophage cocktail (designated ECP-100) containing three Myoviridae phages lytic for Escherichia coli O157:H7 was examined for its ability to reduce experimental contamination of hard surfaces (glass coverslips and gypsum boards), tomato, spinach, broccoli, and ground beef by three virulent strains of the bacterium. The hard surfaces and foods contaminated by a mixture of three E. coli O157:H7 strains were treated with ECP-100 (test samples) or sterile phosphate-buffered saline buffer (control samples), and the efficacy of phage treatment was evaluated by comparing the number of viable E. coli organisms recovered from the test and control samples. Treatments (5 min) with the ECP-100 preparation containing three different concentrations of phages (1010, 109, and 108 PFU/ml) resulted in statistically significant reductions (P = <0.05) of 99.99%, 98%, and 94%, respectively, in the number of E. coli O157:H7 organisms recovered from the glass coverslips. Similar treatments resulted in reductions of 100%, 95%, and 85%, respectively, in the number of E. coli O157:H7 organisms recovered from the gypsum board surfaces; the reductions caused by the two most concentrated phage preparations were statistically significant. Treatment with the least concentrated preparation that elicited significantly less contamination of the hard surfaces (i.e., 109 PFU/ml) also significantly reduced the number of viable E. coli O157:H7 organisms on the four food samples. The observed reductions ranged from 94% (at 120 ± 4 h posttreatment of tomato samples) to 100% (at 24 ± 4 h posttreatment of spinach samples). The data suggest that naturally occurring bacteriophages may be useful for reducing contamination of various hard surfaces, fruits, vegetables, and ground beef by E. coli O157:H7.


Genetics ◽  
1988 ◽  
Vol 119 (3) ◽  
pp. 485-490
Author(s):  
L L Parker ◽  
B G Hall

Abstract Escherichia coli K12 is being used to study the potential for adaptive evolution that is present in the genome of a single organism. Wild-type E. coli K12 do not utilize any of the beta-glucoside sugars arbutin, salicin or cellobiose. It has been shown that mutations at three cryptic loci allow utilization of these sugars. Mutations in the bgl operon allow inducible growth on arbutin and salicin while cel mutations allow constitutive utilization of cellobiose as well as arbutin and salicin. Mutations in a third cryptic locus, arbT, allow the transport of arbutin. A salicin+ arbutin+ cellobiose+ mutant has been isolated from a strain which is deleted for the both the bgl and cel operons. Because the mutant utilized salicin and cellobiose as well as arbutin, it is unlikely it is the result of a mutation in arbT. A second step mutant exhibited enhanced growth on salicin and a third step mutant showed better growth on cellobiose. A fourfold level of induction in response to arbutin and a twofold level of induction in response to salicin was observed when these mutants were assayed on the artificial substrate p-nitrophenyl-beta-D-glucoside. Although growth on cellobiose minimal medium can be detected after prolonged periods of time, these strains are severely inhibited by cellobiose in liquid medium. This system has been cloned and does not hybridize to either bgl or cel specific probes. We have designated this gene system the sac locus. The sac locus is a fourth set of genes with the potential for evolving to provide beta-glucoside utilization.


1999 ◽  
Vol 65 (9) ◽  
pp. 4276-4279 ◽  
Author(s):  
Kumar S. Venkitanarayanan ◽  
Gabriel O. Ezeike ◽  
Yen-Con Hung ◽  
Michael P. Doyle

ABSTRACT The efficacy of electrolyzed oxidizing water for inactivatingEscherichia coli O157:H7, Salmonella enteritidis, and Listeria monocytogenes was evaluated. A five-strain mixture of E. coli O157:H7,S. enteritidis, or L. monocytogenes of approximately 108 CFU/ml was inoculated in 9 ml of electrolyzed oxidizing water (treatment) or 9 ml of sterile, deionized water (control) and incubated at 4 or 23°C for 0, 5, 10, and 15 min; at 35°C for 0, 2, 4, and 6 min; or at 45°C for 0, 1, 3, and 5 min. The surviving population of each pathogen at each sampling time was determined on tryptic soy agar. At 4 or 23°C, an exposure time of 5 min reduced the populations of all three pathogens in the treatment samples by approximately 7 log CFU/ml, with complete inactivation by 10 min of exposure. A reduction of ≥7 log CFU/ml in the levels of the three pathogens occurred in the treatment samples incubated for 1 min at 45°C or for 2 min at 35°C. The bacterial counts of all three pathogens in control samples remained the same throughout the incubation at all four temperatures. Results indicate that electrolyzed oxidizing water may be a useful disinfectant, but appropriate applications need to be validated.


2009 ◽  
Vol 72 (10) ◽  
pp. 2208-2211 ◽  
Author(s):  
A. M. LAURY ◽  
M. V. ALVARADO ◽  
G. NACE ◽  
C. Z. ALVARADO ◽  
J. C. BROOKS ◽  
...  

The objectives of this study were to determine the effects of a lactic acid– and citric acid–based antimicrobial product on the reduction of Salmonella on whole broiler carcasses during processing and the reduction of Salmonella and Escherichia coli O157:H7 on beef trim. Freshly harvested broiler carcasses were inoculated with an inoculum of Salmonella strains to yield a 105 CFU/ml pathogen load on the surface of the carcass. The beef tips were inoculated as well with an inoculum of either E. coli O157:H7 or Salmonella to yield 104 CFU/100 cm2. After 30 min for attachment, the broiler carcasses were treated with Chicxide applied for 5 s via a spray or immersed in Chicxide for 5, 10, or 20 s. Broiler carcasses were rinsed in poultry rinse bags with 400 ml of Butterfield's phosphate buffer in which Salmonella was enumerated from the diluents and Butterfield's phosphate. Chicxide significantly reduced Salmonella by 1.3 log CFU/ml with spray treatment and 2.3 log CFU/ml for all dip treatments. Following 30 min of attachment, the beef tips were placed into a spray cabinet with either Beefxide or sterilized water (control) and sprayed at 1 ft/2.5 s chain speed at 40 lb/in2. The external surface of each beef tip was swabbed (100 cm2) to determine pathogen loads. Beefxide significantly reduced E. coli O157:H7 by 1.4 log CFU/100 cm2 and Salmonella by 1.1 log CFU/100 cm2 (P &lt; 0.05) compared with the control samples.


2006 ◽  
Vol 50 (4) ◽  
pp. 1425-1432 ◽  
Author(s):  
Vasanthi Ramachandran ◽  
B. Chandrakala ◽  
Vidya P. Kumar ◽  
Veeraraghavan Usha ◽  
Suresh M. Solapure ◽  
...  

ABSTRACTClass A high-molecular-weight penicillin-binding protein 1a (PBP1a) and PBP1b ofEscherichia colihave both transglycosylase (TG) and transpeptidase (TP) activity. These enzymes are difficult to assay, since their substrates are difficult to prepare. We show the activity of PBP1a or PBP1b can be measured in membranes by cloning the PBP into anE. coli ponB::Spcrstrain. Using this assay, we show that PBP1a is ∼10-fold more sensitive to penicillin than PBP1b and that the 50% inhibitory concentration (IC50) of moenomycin, a TG inhibitor, is ∼10-fold higher in the PBP transformants than in wild-type membranes; this increase in IC50in transformants can be used to test the specificity of test compounds for inhibition of the TG. Alternatively, the coupled TG-TP activity of PBP1b can be directly measured in a two-step microplate assay. In the first step, radiolabeled lipid II, the TG substrate, was made in membranes of theE. coli ponB::Spcrstrain by incubation with the peptidoglycan sugar precursors. In the second step, the TG-TP activity was assayed by adding a source of PBP1b to the membranes. The coupled TG-TP activity converts lipid II to cross-linked peptidoglycan, which was specifically captured by wheat germ agglutinin-coated scintillation proximity beads in the presence of 0.2% Sarkosyl (B. Chandrakala et al., Antimicrob. Agents Chemother.48:30-40, 2004). The TG-TP assay was inhibited by penicillin and moenomycin as expected. Surprisingly, tunicamycin and nisin also inhibited the assay, and paper chromatography analysis revealed that both inhibited the transglycosylase. The assay can be used to screen for novel antibacterial agents.


2014 ◽  
Vol 59 (1) ◽  
pp. 331-337 ◽  
Author(s):  
Asha S. Nayar ◽  
Thomas J. Dougherty ◽  
Folkert Reck ◽  
Jason Thresher ◽  
Ning Gao ◽  
...  

ABSTRACTIn a previous report (T. J. Dougherty, A. Nayar, J. V. Newman, S. Hopkins, G. G. Stone, M. Johnstone, A. B. Shapiro, M. Cronin, F. Reck, and D. E. Ehmann, Antimicrob Agents Chemother 58:2657–2664, 2014), a novel bacterial type II topoisomerase inhibitor, NBTI 5463, with activity against Gram-negative pathogens was described. First-step resistance mutations inPseudomonas aeruginosaarose exclusively in thenfxBgene, a regulator of the MexCD-OprJ efflux pump system. The present report describes further resistance studies with NBTI 5463 in bothPseudomonas aeruginosaandEscherichia coli. Second-step mutations inP. aeruginosaarose at aspartate 82 of the gyrase A subunit and led to 4- to 8-fold increases in the MIC over those seen in the parental strain with a first-stepnfxBefflux mutation. A third-step mutant showed additional GyrA changes, with no changes in topoisomerase IV. Despite repeated efforts, resistance mutations could not be selected inE. coli. Genetic introduction of the Asp82 mutations observed inP. aeruginosadid not significantly increase the NBTI MIC inE. coli. However, with the aspartate 82 mutation present, it was possible to select second-step mutations in topoisomerase IV that did lead to MIC increases of 16- and 128-fold. As with the gyrase aspartate 82 mutation, the mutations in topoisomerase IV did not by themselves raise the NBTI MIC inE. coli. Only the presence of mutations in both targets ofE. coliled to an increase in NBTI MIC values. This represents a demonstration of the value of balanced dual-target activity in mitigating resistance development.


1998 ◽  
Vol 42 (4) ◽  
pp. 959-962 ◽  
Author(s):  
Michael R. Paradise ◽  
Gregory Cook ◽  
Robert K. Poole ◽  
Philip N. Rather

ABSTRACT The aarE1 allele was identified on the basis of the resulting phenotype of increased aminoglycoside resistance. TheaarE1 mutation also resulted in a small-colony phenotype and decreased levels of aac(2′)-Ia mRNA. The deduced AarE gene product displayed 61% amino acid identity to theEscherichia coli UbiA protein, an octaprenyltransferase required for the second step of ubiquinone biosynthesis. Complementation experiments in both Providencia stuartiiand E. coli demonstrated that aarE andubiA are functionally equivalent.


1999 ◽  
Vol 62 (8) ◽  
pp. 850-856 ◽  
Author(s):  
PETER J. TAORMINA ◽  
LARRY R. BEUCHAT

The behavior of Escherichia coli O157:H7 on alfalfa seeds subjected to conditions similar to those used commercially to grow and market sprouts as it is affected by applications of NaOCl, Ca(OCl)2, acidified NaClO2, acidified ClO2, Na3PO4, Vegi-Clean, Tsunami, Vortexx, or H2O2 at various stages of the sprouting process was determined. Application of 2,000 ppm of NaOCl, 200 and 2,000 ppm of Ca(OCl)2, 500 ppm of acidified ClO2, 10,000 ppm of Vegi-Clean, 80 ppm of Tsunami, or 40 and 80 ppm of Vortexx to germinated seeds significantly reduced the population of E. coli O157:H7. With the exception of acidified NaOCl2 at 1,200 ppm, spray applications of these chemicals did not significantly reduce populations or control the growth of E. coli O157:H7 on alfalfa sprouts during the sprouting process. Populations of E. coli on alfalfa sprouts peaked at 6 to 7 log10 CFU/g 48 h after initiation of the sprouting process and remained stable despite further spraying with chemicals. The population of E. coli O157:H7 on sprouts as they entered cold storage at 9 ± 2°C remained essentially unchanged for up to 6 days. None of the chemical treatments evaluated was able to eliminate or satisfactorily reduce E. coli O157:H7 on alfalfa seeds and sprouts. Observations on the ability of E. coli O157:H7 to grow during production of alfalfa sprouts not subjected to chemical treatments are similar to those from a previous study in our laboratory on the behavior of Salmonella Stanley. Our results do not reveal a chemical treatment method to eliminate the pathogen from alfalfa sprouts. We have demonstrated that currently recommended procedures for sanitizing alfalfa seeds fail to eliminate E. coli O157:H7 and that the pathogen can grow to populations exceeding 7 log10 CFU/g of sprouts produced using techniques not dissimilar to those used in the sprout industry.


1999 ◽  
Vol 62 (11) ◽  
pp. 1336-1340 ◽  
Author(s):  
MINDY M. BRASHEARS ◽  
WENDY A. DURRE

Cells of Lactobacillus lactis were added to trypticase soy broth that contained cells of Escherichia coli O157:H7 or cells of Salmonella spp. in order to determine if L. lactis inhibited the pathogens. The inhibition of all pathogens was examined during growth at 37°C for 24 h. Inhibition of Salmonella spp. was also examined at refrigeration temperatures (6°C) for 5 days. One strain each of E. coli O157:H7, Salmonella Typhimurium, and Salmonella Enteritidis was examined. E. coli was enumerated on violet red bile agar, and Salmonella spp. were enumerated on brilliant green agar. In all experiments at 37°C, the L. lactis completely inhibited all pathogens, producing numbers that were not detectable after 24 h of incubation. There were significant (P &gt; 0.05) increases in numbers of the pathogens in the control samples containing no L. lactis. There were significant (P &lt; 0.05) declines in the pH of both control and L. lactis inoculated samples. There was a significantly (P &lt; 0.05) larger decline in the pH of samples inoculated with L. lactis. Interaction studies with pH-neutralized broth indicated that acid production by L. lactis was primarily responsible for the inhibition. Numbers of Salmonella spp. incubated at 6°C did not decline significantly (P &gt; 0.05) for control or inoculated samples, which suggests that this strain of L. lactis does not inhibit Salmonella spp. at refrigeration temperatures. Additionally, there were no significant (P &gt; 0.05) changes in pH or in numbers of L. lactis during refrigerated storage.


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