Diversity and ecological function of urease-producing bacteria in the cultivation environment of Gracilariopsis lemaneiformis.

Urease-producing bacteria (UPB) provide inorganic nitrogen for primary producers by hydrolyzing urea, and play an important role in marine nitrogen cycle. However, there is still an incomplete understanding of UPB and their ecological functions in the cultivation environment of the red macroalgae Gracilariopsis lemaneiformis. This study comprehensively analyzed the diversity of culturable UPB and explored their effects on urea uptake by G. lemaneiformis. A total of 34 isolates belonging to four main bacterial phyla i.e. (Proteobacteria, Bacteroidetes, Firmicutes, and Actinobacteria) were identified through 16S rRNA sequencing and were screened for UPB by urea agar chromogenic medium assay and ureC gene cloning. Our data revealed that only 8 strains contained urease. All of these UPB exhibited different urease activities, which were determined by the Berthelot reaction colorimetry assay. Additionally, the UPB strain (G13) isolated from G. lemaneiformis with higher urease activity was selected for co-culture with G. lemaneiformis to explore its role in promoting or inhibiting nitrogen uptake by macroalgae. The results showed a significant increase in urea consumption in the culture medium and the total cellular nitrogen in G. lemaneiformis in the UPB-co culture group compared to the sterile group. This suggests that the selected UPB strain positively influences nitrogen uptake by G. lemaneiformis. Similarly, isotopic assays revealed that the δ15N content of G. lemaneiformis was significantly higher in the UPB-co culture than in the control group, where δ15N-urea was the only nitrogen source in the culture medium. This indicates that the UPB helped G. lemaneiformis to absorb more nitrogen from urea. Moreover, the highest content of δ15N was found in G. lemaneiformis with epiphytic bacteria compared to sterilized (i.e. control), showing that epiphytic bacteria, along with UPB, have a compound effect in helping G. lemaneiformis absorb more nitrogen from urea. Taken together, these results provide unique insight into the ecological role of UPB and suggest that urease from macroalgae environment-associated bacteria might be an important player in marine nitrogen cycling.

Urease-producing bacteria with plant growth-promoting ability that may tolerate and remove cadmium from aqueous solution.

Urease-producing bacteria (UPB) are widely present in soil and play an important role in soil ecosystems. In this study, 65 UPB strains were isolated from cadmium (Cd)-polluted soil around a lead-zinc mine in Yunnan Province, China. The Cd tolerance, removal of Cd from aqueous solution, production of indoleacetic acid (IAA) and plant growth-promoting effects of these materials were investigated. The results indicate that among the 65 UPB strains, four strains with IAA-producing ability were screened and identified as Bacillus thuringiensis W6-11, B. cereus C7-4, Serratia marcescens W11-10, and S. marcescens C5-6. Among the four strains, B. cereus C7-4 had the highest Cd tolerance, median effect concentration (EC50) of 59.94 mg/L. Under Cd 5 mg/L, S. marcescens C5-6 had the highest Cd removal from aqueous solution, up to 69.83%. Under Cd 25 mg/kg, inoculation with B. cereus C7-4 significantly promoted maize growth in a sand pot by increasing the root volume, root surface area, and number of root branches by 22%, 29%, and 20%, respectively, and plant height and biomass by 16% and 36%, respectively, and significantly increasing Cd uptake in the maize roots. Therefore, UPB is a potential resource for enhancing plant adaptability to Cd stress in plants with Cd-polluted habitats.

This study utilized urease-producing bacteria screened from the soil of lead zinc mining areas in Yunnan, China as the research object, enriching the microbial resources in Yunnan. In addition, this article verified the IAA production ability and cadmium removal ability of urease-producing bacteria, and screened out bifunctional urease-producing bacteria that have potential in cadmium pollution control and plant growth promotion.

Heavy metals remediation through bio-solidification: Potential application in environmental geotechnics.

Heavy metals are pervasive pollutants found in water, soil, and solid wastes. Bio-solidification offers an environmentally friendly approach to immobilize heavy metal ions using two types of bacteria: urease-producing bacteria (UPB) and phosphatase-producing bacteria (PPB). UPB, exemplified by Sporosarcina pasteurii, secretes urease to hydrolyze urea and generate CO32- ions, while PPB, like Bacillus subtilis, produces alkaline phosphatase to hydrolyze organophosphate monoester (ROP) and produce PO43- ions. These ions react with heavy metal ions, effectively reducing their concentration by forming insoluble carbonate or phosphate precipitates. The success of bio-solidification is influenced by various factors, including substrate concentration, temperature, pH, and bacterial density. Optimal operational conditions can significantly enhance the remediation performance of heavy metals. UPB and PPB hold great potential for remediating heavy metal pollution in diverse contaminated areas such as tailings ponds, electroplating sewage, and garbage incineration plants. In conclusion, harnessing the power of these microbial methods can provide effective solutions for remediating heavy metal-induced pollution across a range of environmental conditions.

Effects of water activity, ammonia and Corynebacterium urealyticum on the survival of Salmonella Typhimurium in sterile poultry litter.

AIMS: This study examined the effects of water activity (aw ), ammonia and Corynebacterium urealyticum on the survival of Salmonella Typhimurium in sterile poultry litter. METHODS AND RESULTS: Sterile poultry litter inoculated with S. Typhimurium was adjusted to pH 9.0, various aw levels (0.84, 0.92 and 0.96), and total ammonia nitrogen levels were increased either by the addition of ammonium sulphate or C. urealyticum inoculation with 1% urea added. All litter treatments were incubated at 30°C and sampled daily for five days. Similar results were observed at each aw level in both experiments. At 0.84 and 0.92 aw , S. Typhimurium populations in litter fell below 1 log CFU g-1 within 5 days, with no significant differences between the controls and increased ammonia treatments. At 0.96 aw , Salmonella populations treated with increased ammonia levels were significantly lower than control treatments on days 1-5. CONCLUSIONS: This study showed that C. urealyticum can produce ammonia in litter at higher aw levels with sufficient available urea and that the antimicrobial efficacy of ammonia is dependent on high aw (~0.96) in litter. SIGNIFICANCE AND IMPACT OF THE STUDY: These results provide insights into the production of ammonia in litter, its antimicrobial efficacy in litter and the importance of aw in this interaction.

A quantitative, high-throughput urease activity assay for comparison and rapid screening of ureolytic bacteria.

Urease is a dinickel enzyme commonly found in numerous organisms that catalyses the hydrolysis of urea into ammonia and carbon dioxide. The microbially induced carbonate precipitation (MICP) process mediated by urease-producing bacteria (UPB) can be used for many applications including, environmental bioremediation, soil improvement, healing of cracks in concrete, and sealing of rock joints. Despite the importance of urease and UPB in various applications, a quantitative, high-throughput assay for the comparison of urease activity in UPB and rapid screening of UPB from diverse environments is lacking. Herein, we reported a quantitative, 96-well plate assay for urease activity based on the Christensen's urea agar test. Using this assay, we compared urease activity of six bacterial strains (E. coli BL21, P. putida KT2440, P. aeruginosa PAO1, S. oneidensis MR-1, S. pasteurii DSM 33, and B. megaterium DSM 319) and showed that S. pasteurii DSM 33 exhibited the highest urease activity. We then applied this assay to quantify the inhibitory effect of calcium on urease activity of S. pasteurii DSM 33. No significant inhibition was observed in the presence of calcium at concentrations below 10 mM, while the urease activity decreased rapidly at higher concentrations. At a concentration higher than 200 mM, calcium completely inhibited urease activity under the tested conditions. We further applied this assay to screen for highly active UPB from a wastewater enrichment and identified a strain of S. pasteurii exhibiting a substantially higher urease activity than DSM 33. Taken together, we established a 96-well plate-based quantitative, high-throughput urease activity assay that can be used for comparison and rapid screening of UPB. As UPB and urease activity are of interest to environmental, civil, and medical researchers and practitioners, we envisage wide applications of the assay reported in this study.

Hyperammonemic encephalopathy associated with post-radiotherapy vesicointestinal fistula following cervical cancer.

The majority of hyperammonemic encephalopathy is due to liver cirrhosis. However, urinary tract infection caused by urease-producing bacteria increases ammonia in urine and can lead to hyperammonemic encephalopathy, especially in cases with obstructive uropathy and vesicointestinal fistula. This is the first case report of hyperammonemic encephalopathy in a cervical cancer patient associated with postradiotherapy vesicointestinal fistula. A 52-year-old woman developed diarrhea due to vesicosigmoidal fistula 14 years after radical hysterectomy and radiotherapy to treat cervical cancer. She refused urinary and/or fecal diversion. Twelve months after the diagnosis of fistula, she was admitted due to somnolence. Blood examination showed hyperammonemia without liver dysfunction. Urine culture showed Proteus rettgeri and Klebsiella pneumoniae. She recovered after intravenous antibiotics. Eight months after recovery, she was readmitted due to somnolence reoccurring with failed intravenous, but successful oral antibiotic treatment. She finally agreed to undergo percutaneous nephrostomy and hyperammonemia never recurred during 7 years of follow-up.

[A case of hyperammonemia caused by urinary tract infection due to urease-producing bacteria in dementia with Lewy bodies].

An 87-year-old woman diagnosed with dementia with Lewy bodies (DLB) 2 years earlier was referred to our institution because of difficulty walking. She was diagnosed with urinary tract infection and admitted to our hospital. During hospitalisation, she became delirious, which prompted the administration of haloperidol. Afterwards, an altered level of consciousness was noted, measuring 300 on the Japan coma scale. A blood test revealed hyperammonaemia without liver damage. Urine culture detected the presence of Corynebacterium urealyticum. Therefore, we diagnosed this case as one of hyperammonaemia due to urinary tract infection caused by urease-producing bacteria. Soon after the insertion of a urethral catheter, the ammonia level decreased, and the consciousness level improved. In this case, the patient took medication to preserve her bladder function, which is frequently associated with DLB. We suspected that the drug caused urinary retention, resulting in hyperammonaemia. Hyperammonaemia due to these bacteria should be considered in DLB patients with an impaired consciousness, especially in those using regulators of the urinary bladder function.

Environmental safety and biosafety in construction biotechnology.

The topics of Construction Biotechnology are the development of construction biomaterials and construction biotechnologies for soil biocementation, biogrouting, biodesaturation, bioaggregation and biocoating. There are known different biochemical types of these biotechnologies. The most popular construction biotechnology is based on precipitation of calcium carbonate initiated by enzymatic hydrolysis of urea which follows with release of ammonia and ammonium to environment. This review focuses on the hazards and remedies for construction biotechnologies and on the novel environmentally friendly biotechnologies based on precipitation of hydroxyapatite, decay of calcium bicarbonate, and aerobic oxidation of calcium salts of organic acids. The use of enzymes or not living bacteria are the best options to ensure biosafety of construction biotechnologies. Only environmentally safe construction biotechnologies should be used for such environmental and geotechnical engineering works as control of the seepage in dams, channels, landfills or tunnels, sealing of the channels and the ponds, prevention of soil erosion and soil dust emission, mitigation of soil liquefaction, and immobilization of soil pollutants.

[A case of hyperammonemia resulting from urinary tract infection caused by urease-producing bacteria in a Parkinson's disease patient with drug-induced urinary retention].

A 71-year-old woman with a 9-year history of Parkinson's disease was admitted to our hospital emergently because of consciousness disturbance. Her consciousness level was 200 on the Japan coma scale (JCS), and she presented with tenderness and distension of the lower abdomen. Brain computed tomography showed normal findings. Blood tests showed an increased ammonia level (209 µg/dl) with normal AST and ALT levels. We catheterized the bladder for urinary retention. Five hours after admission, the blood ammonia level decreased to 38 µg/dl, and her consciousness level improved dramatically. Corynebacterium urearyticum, a bacterial species that produces urease, was detected by urine culture. Therefore, she was diagnosed with hyperammonemic encephalopathy resulting from urinary tract infection caused by urease-producing bacteria. In this case, urologic active agents had been administered to treat neurogenic bladder. We suspect that these drugs caused urinary obstruction and urinary tract infection. It is important to recognize that obstructive urinary tract infection caused by urease-producing bacteria can cause hyperammonemia. Neurological disorders, such as Parkinson's disease, tend to complicate neurogenic bladder. This disease should be considered in elderly patients with Parkinson's disease who are receiving urologic active drugs.

In vitro screening and evaluation of some Indian medicinal plants for their potential to inhibit Jack bean and bacterial ureases causing urinary infections.

CONTEXT: Bacterial ureases play an important role in pathogenesis of urinary infections. Selection of plants was done on the basis of their uses by the local people for the treatment of various bacterial and urinary infections. OBJECTIVE: Our investigation screens and evaluates 15 Indian medicinal plants for their possible urease inhibitory activity as well as their ability to inhibit bacteria causing urinary infections. MATERIALS AND METHODS: Plant extracts in three different solvents (methanol, aqueous, and cow urine) were screened for their effect on Jack-bean urease using the phenol-hypochlorite method. Subsequently, seven bacterial strains were screened for their ability to release urease and further antimicrobial-linked urease inhibition activity and minimum inhibitory concentration of the tested extracts were evaluated by the agar well diffusion and microdilution method, respectively. RESULTS: Five plants out of 15 crude extracts revealed good urease inhibitory activity (≥ 20% at 1 mg/ml conc.) and IC50 values for these extracts ranged from 2.77 to 0.70 mg/ml. Further testing of these extracts on urease-producing bacterial strains (Staphylococcus aureus NCDC 109, S. aureus MTCC 3160, Proteus vulgaris MTCC 426, Klebsiella pneumoniae MTCC 4030, and Pseudomonas aeruginosa MTCC 7453) showed good anti-urease potency with an MIC ranging from 500 to 7.3 µg/ml. DISCUSSION AND CONCLUSION: The results of screening as well as susceptibility assay clearly revealed a strong urease inhibitory effect of Acacia nilotica L. (Fabaceae), Emblica officinalis Gaertn. (Phyllanthaceae), Psidium guajava L. (Myrtaceae), Rosa indica L. (Rosaceae), and Terminalia chebula Retz. (Combretaceae). Our findings may help to explain the beneficial effect of these plants against infections associated with the urease enzyme.

Urease-producing bacteria combined with pig manure biochar immobilize Cd and inhibit the absorption of Cd in lettuce (Lactuca sativa L.).

The synergistic remediation of heavy metal-contaminated soil by functional strains and biochar has been widely studied. However, the mechanisms by which urease-producing bacteria combine with pig manure biochar (PMB) to immobilize Cd and inhibit Cd absorption in vegetables are still unclear. In our study, the effects and mechanisms of PMB combined with the urease-producing bacterium TJ6 (TJ6 + PMB) on Cd adsorption were explored. The effects of TJ6 + PMB on the Cd content and pH of the leachate were also studied through a 56-day soil leaching experiment. Moreover, the effects of the complexes on Cd absorption and microbial mechanisms in lettuce were explored through pot experiments. The results showed that PMB provided strain TJ6 with a greater ability to adsorb Cd, inducing the generation of CdS and CdCO3, and thereby reducing the Cd content (71.1%) and increasing the pH and urease activity in the culture medium. TJ6 + PMB improved lettuce dry weight and reduced Cd absorption. These positive effects were likely due to (1) TJ6 + PMB increased the organic matter and NH4+ contents, (2) TJ6 + PMB transformed available Cd into residual Cd and decreased the Cd content in the leachate, and (3) TJ6 + PMB altered the structure of the rhizosphere bacterial and fungal communities in lettuce, increasing the relative abundances of Stachybotrys, Agrocybe, Gaiellales, and Gemmatimonas. These genera can promote plant growth, decompose organic matter, and release phosphorus. Interestingly, the fungal communities were more sensitive to the addition of TJ6 and PMB, which play important roles in the decomposition of organic matter and immobilization of Cd. In conclusion, this study revealed the mechanism by which urease-producing bacteria combined with pig manure biochar immobilize Cd and provided a theoretical basis for safe pig manure return to Cd-polluted farmland. This study also provides technical approaches and bacterial resources for the remediation of heavy metal-contaminated soil.

Association between urinary lithiasis, other than struvite by crystallography and non-ureolytic bacteria.

The relationship between urinary tract infection caused by urease-producing bacteria and lithiasis due to struvite stones is well established in the literature. However, there is limited knowledge on whether non-urease producing bacteria can also promote crystallization. In our study, we analyzed the association between urinary lithiasis, other than struvite by crystallography and non-ureolytic bacteria, in 153 patients who underwent surgery for urinary stone. The collected samples were sent for crystallographic analysis and culture. Additionally, preoperatory urine culture was collected for combined evaluation with the previous data. Percutaneous nephrolithotomy was the most commonly performed approach (45.8%). Struvite stones were more frequently identified in women (90.3%). Among stones with positive cultures, except struvite, 45.5% were composed of calcium oxalate monohydrate. The difference between urine culture and stone culture was different in 24.8% of the cases. Among stones with positive cultures that did not contain struvite, 86.4% showed non-urease bacteria in their cultures and 47.1% of struvite stones also did not have urease-producing bacteria in their cultures (p < 0.021). Our findings suggest that there is an association between non-ureolytic bacteria and stones that are not composed of struvite.

Inhibitory effect of polyphenols from sumac, pomegranate and Indian almond on urease producing bacteria and jack bean urease activity.

Urinary tract infection caused by Klebsiella, Proteus and Streptococcus is a urease dependent process, hence treatment of these infections by antibacterial compounds lies in inhibition of their virulence factors. The crude methanolic extracts derived from sumac fruit, pomegranate peel and Indian almond leaves were separated into anthocyanin and non-anthocyanin fractions using solid phase cartridges. The inhibitory effect of these fractions was determined on the growth of urease producing species and jack bean urease activity. Known compounds in the fractions were also docked with ureases of different biological origins viz. K. pneumoniae (PDB ID: 8HCN), K. aerogenes (PDB ID: 2KAU), Helicobacter pylori (PDB ID:8HC1)and Canavalia ensiformis (jack bean) (PDB ID: 3LA4) to determine their binding affinities and interaction with the enzyme. All the fractions showed significant inhibition growth for P. mirabilis, S. epidermidis and K.pneumoniae. Among the samples, sumac showed greatest inhibition against all (MIC 6-25 mg.mL-1) while among the fractions, anthocyanin was found to be most active (MIC 6-12 mg/mL). Likewise, all fractions inhibited urease with lowest ICs50 shown by sumac fractions (21-116 µg.mL-1). Out of 39 compounds docked, 27 showed interaction with movable flaps and/or active site of ureases which explains their mode of inhibition.

A Diabetic Patient with Prolonged Hyperammonemia Due to Urinary Tract Infection Caused by Urease-producing Bacteria.

Obstructive tract infection caused by urease-producing bacteria can lead to hyperammonemia. We herein report a 73-year-old woman with diabetes and compensated liver cirrhosis who developed obstructive pyelonephritis. Her consciousness level declined on day 3 of hospitalization. Blood tests revealed an elevated ammonia level of 140 µg/dL. Urine and blood cultures identified Proteus mirabilis as a urease-producing bacterium. The treatment included double J (WJ) catheter insertion and administration of antimicrobial agents. On day 7 of hospitalization, the ammonia level gradually decreased, and the patient's consciousness improved. However, despite these interventions, perinephric overflow of urine persisted, necessitating nephrectomy.

Recurrent Non-cirrhotic Hyperammonemic Encephalopathy Due to Complicated Urinary Tract Infection: A Case Report.

Hyperammonemic encephalopathy (HE) can be broadly defined as an alteration in the level of consciousness due to elevated blood ammonia level. While hepatic cirrhosis is the most common cause of HE, non-hepatic causes like drugs, infections, and porto-systemic shunts can also lead to the presentation. In this case, we highlight an unusual occurrence of recurrent non-cirrhotic HE from obstructive urinary tract infection (UTI) with urea-splitting micro-organisms in an elderly male patient. The patient exhibited altered mentation, and elevated ammonia levels with normal hepatic function at presentation. Urine culture revealed Proteus mirabilis resistant to extended spectrum beta-lactamases (ESBL). Successful management of obstructive UTI was achieved through Foley's catheterization and intravenous (IV) antibiotics, resulting in the resolution of HE. This outcome further supports the significance of UTI as a potential cause of hyperammonemia. Thus, UTI as one of the non-hepatic causes of hyperammonemia should always be explored among elderly patients presenting with altered mentation.

Hyperammonemic Encephalopathy Secondary to Urinary Tract Infection.

Hyperammonemic encephalopathy (HE) refers to a clinical condition characterized by abrupt alteration in mental status (AMS) with markedly elevated plasma ammonia levels and frequently results in intractable coma and death. While hepatic cirrhosis is by far the most common etiology for hyperammonemia together with drugs such as valproic acid as well as urea cycle disorders, non-hepatic causes of hyperammonemia are rare and pose a clinical challenge. In this report, we describe a case of HE caused by obstructive urinary tract infection due to urease-producing bacteria in a 69-year-old man with two episodes of obstructive uropathy associated with AMS resolving with treatment with antibiotics and lactulose with normalization of ammonia level. We also provide a review of the literature with emphasis on the recognition of this serious entity of HE in the setting of obstructive uropathy to avoid the possible complications that include intractable coma and high mortality from this potentially treatable disorder.

Inhibition of cadmium uptake by wheat with urease-producing bacteria combined with sheep manure under field conditions.

In heavy metal-contaminated farmland, microorganisms or organic fertilizers can be used to minimize heavy metal uptake by crops to ensure food safety. However, the mechanisms by which urease-producing and metal-immobilizing bacteria combined with manure inhibit Cd uptake in wheat (Triticum aestivum L.) remain unclear. Herein, the effects of Enterobacter bugandensis TJ6, sheep manure (SM), and TJ6 combined with SM on Cd uptake by wheat and the mechanisms involved were investigated under field conditions. The results showed that strain TJ6 increased the urease activity and the proportion of strains with a high Cd adsorption capacity in SM, thereby enhancing the Cd adsorption capacity of SM in solution. Strain TJ6 combined with SM improved the rhizosphere soil urease activity, NH4+/NO3- ratio, and pH, thus reducing the Cd content (75.9%) in wheat grain. In addition, TJ6+SM reduced the bacterial community diversity but shifted the structure of the bacterial community in rhizosphere soil. Interestingly, the relative abundances of urease-producing bacteria and metal-immobilizing bacteria (Enterobacter, Bacillus, Exiguobacterium, Rhizobium, and Serratia) in rhizosphere soil were enriched, which enhanced wheat resistance to Cd toxicity. These results showed that urease-producing and metal-immobilizing bacteria combined with sheep manure can inhibit the uptake of Cd by wheat.

Effects of urease-producing bacteria and eggshell on physiological characteristics and Cd accumulation of pakchoi (Brassica chinensis L.) plants.

Soil cadmium (Cd) contamination resulting from anthropogenic activity poses severe threats to food safety and human health. In this study, a pot experiment was performed to evaluate the possibility of using urease-producing bacterium UR21 and eggshell (ES) waste for improving the physiological characteristics and reducing Cd accumulation of pakchoi (Brassica chinensis L.) plants. UR21 has siderophore and IAA production ability. The application of UR21 and ES individually or in combination could improve the root and shoot length, and fresh and dry weight of pakchoi plants under Cd stress. In Cd + ES + UR21-treated plants, the dry weight of shoot and root were increased by 61.54% and 72.73%, respectively. The chlorophyll a, chlorophyll b, and carotenoid content were increased by 52.19%, 42.95%, and 95.56% in Cd + ES + UR21-treated plants. Meanwhile, the H2O2 and MDA content were decreased while the SOD and POD activity were increased, and an increase of soluble protein level in pakchoi plants was observed under Cd + ES + UR21 treatment. Importantly, eggshell and UR21 alone or in combination induced a decline of Cd content in pakchoi plants, especially that Cd + ES + UR21 treatment decreased Cd content in shoot and root by 26.96% and 42.91%, respectively. Meanwhile, the soil urease and sucrase activities were enhanced. Generally, the combined application of ureolytic bacteria UR21 and eggshell exhibited better effects than applied them individually in terms of alleviating Cd toxicity in pakchoi plants. Our findings may give a unique perspective for an eco-friendly and sustainable strategy to remediate heavy metal-polluted soils.

Mechanical properties of bio-cementation materials in pre-precipitation mixing process.

Urease-producing bacteria (UPB) could be used to cement loose sand particles. The UPB would produce free ammonia and carbon dioxide during the process of hydrolyzing urea, and part of the free ammonia would be discharged into the air to cause certain pollution to the atmospheric environment. The carbon dioxide could react with alkaline oxide to form carbonates and improved the strength in GGBS comparing with medium containing different concentrations of urea. By adding hydrogen phosphate ions and magnesium salts, free ammonia could be converted into environmentally friendly magnesium ammonium phosphate. The mixture of biological magnesium ammonium phosphate and basic magnesium carbonate could be synthesized through the bio-mineralization process. Through the pre-precipitation mixing process, the loose sand particles could be cemented into a whole. Scanning electron microscopy (SEM) images of the sand column showed that the mixture of biological magnesium ammonium phosphate and basic magnesium carbonate could better fill in the pores of sand grains. In the pre-precipitation mixing process, the optimal standing time and dosage of the bio-cement slurry prepared by the bio-mineralization method were 6 h and 30%, respectively. The average interface bonding force between CJ2 and glass slide was 2.12 N.

Antimicrobial Efficacy of Un-Ionized Ammonia (NH3) against Salmonella Typhimurium in Buffered Solutions with Variable pH, NH3 Concentrations, and Urease-Producing Bacteria.

The presence of Salmonella in poultry litter, when used as a biological soil amendment, presents a risk for the preharvest contamination of fresh produce. Poultry litter is rich in organic nitrogen, and previous studies have suggested that ammonia (NH3) in poultry litter may affect the survival of Salmonella. Salmonella enterica serovar Typhimurium was inoculated into buffer solutions to characterize the pH dependency, minimum antimicrobial concentration, and efficacy of NH3 production. In solutions with 0.4 M total ammonia nitrogen (TAN) at various pH levels (5, 7, 8, and 9), significant inactivation of Salmonella only occurred at pH 9. Salmonella was reduced by ∼8 log CFU/mL within 12 to 18 h at 0.09, 0.18, 0.26, and 0.35 M NH3. The minimum antimicrobial concentration tested was 0.04 M NH3, resulting in an ∼7 log CFU/mL reduction after 24 h. Solutions with urea (1% and 2%) and urease enzymes rapidly produced NH3, which significantly reduced Salmonella within 12 h. The urease-producing bacterium Corynebacterium urealyticum showed no antagonistic effects against Salmonella in solution. Conversely, with 1% urea added, C. urealyticum rapidly produced NH3 in solution and significantly reduced Salmonella within 12 h. Salmonella inactivation data were nonlinear and fitted to Weibull models (Weibull, Weibull with tailing effects, and double Weibull) to describe their inactivation kinetics. These results suggest that high NH3 levels in poultry litter may reduce the risk of contamination in this biological soil amendment. This study will guide future research on the influence of ammonia on the survival and persistence of Salmonella in poultry litter. IMPORTANCE Poultry litter is a widely used biological soil amendment in the production of fresh produce. However, poultry litter may contain human pathogens, such as Salmonella, which introduces the risk of preharvest produce contamination in agricultural fields. Ammonia in poultry litter, produced through bacterial degradation of urea, may be detrimental to the survival of Salmonella; however, these effects are not fully understood. This study utilized aqueous buffer solutions to demonstrate that the antimicrobial efficacy of ammonia against Salmonella is dependent on alkaline pH levels, where increasing concentrations of ammonia led to more rapid inactivation. Inactivation was also demonstrated in the presence of urea and urease or urease-producing Corynebacterium urealyticum. These findings suggest that high levels of ammonia in poultry litter may reduce the risk of contamination in biological soil amendments and will guide further studies on the survival and persistence of Salmonella in poultry litter.