Understanding the role of zinc ions on struvite nucleation and growth in the context of infection urinary stones.

Taking into account that in recent decades there has been an increase in the incidence of urinary stones, especially in highly developed countries, from a wide range of potentially harmful substances commonly available in such countries, we chose zinc for the research presented in this article, which is classified by some sources as a heavy metal. In this article, we present the results of research on the influence of Zn2+ ion on the nucleation and growth of struvite crystals-the main component of infection urinary stones. The tests were carried out in an artificial urine environment with and without the presence of Proteus mirabilis bacteria. In the latter case, the activity of bacterial urease was simulated chemically, by systematic addition of an aqueous ammonia solution. The obtained results indicate that Zn2+ ions compete with Mg2+ ions, which leads to the gradual replacement of Mg2+ ions in the struvite crystal lattice with Zn2+ ions to some extent. This means co-precipitation of Mg-struvite (MgNH4PO4·6H2O) and Znx-struvite (Mg1-xZnxNH4PO4·6H2O). Speciation analysis of chemical complexes showed that Znx-struvite precipitates at slightly lower pH values than Mg-struvite. This means that Zn2+ ions shift the nucleation point of crystalline solids towards a lower pH. Additionally, the conducted research shows that Zn2+ ions, in the range of tested concentrations, do not have a toxic effect on bacteria; on the contrary, it has a positive effect on cellular metabolism, enabling bacteria to develop better. It means that Zn2+ ions in artificial urine, in vitro, slightly increase the risk of developing infection urinary stones.

Antibacterial properties and urease suppression ability of Lactobacillus inhibit the development of infectious urinary stones caused by Proteus mirabilis.

Infectious urolithiasis is a type of urolithiasis, that is caused by infections of the urinary tract by bacteria producing urease such as Proteus mirabilis. Lactobacillus spp. have an antagonistic effect against many pathogens by secreting molecules, including organic acids. The aim of the study was to determine the impact of Lactobacillus strains isolated from human urine on crystallization of urine components caused by P. mirabilis by measuring bacterial viability (CFU/mL), pH, ammonia release, concentration of crystallized salts and by observing crystals by phase contrast microscopy. Moreover, the effect of lactic acid on the activity of urease was examined by the kinetic method and in silico study. In the presence of selected Lactobacillus strains, the crystallization process was inhibited. The results indicate that one of the mechanisms of this action was the antibacterial effect of Lactobacillus, especially in the presence of L. gasseri, where ten times less P. mirabilis bacteria was observed, compared to the control. It was also demonstrated that lactic acid inhibited urease activity by a competitive mechanism and had a higher binding affinity to the enzyme than urea. These results demonstrate that Lactobacillus and lactic acid have a great impact on the urinary stones development, which in the future may help to support the treatment of this health problem.

Insights into the physical and chemical properties of struvite crystal surfaces in terms of the effectiveness of bacterial adhesion.

In this paper, we present the results of research on the physicochemical properties of two selected faces of the struvite crystal, which is the main component of infectious urinary stones. Two main faces, (001) and ([Formula: see text]), ending the c-axis, were selected for the study. These faces are not related by symmetry relations, which means, among other things, that they should have a different atomic structure, which was confirmed experimentally. In addition, the studies show that the tested surfaces have hydrophilic properties, however, the ([Formula: see text]) face is more hydrophilic compared to the (001) face. The physicochemical properties of the crystal as a whole, as well as the physicochemical properties of these faces influence the magnitude of adhesion. The adhesive force in both water and artificial urine is greater for face ([Formula: see text]) compared to face (001). The assessment of the adhesion of Proteus mirabilis bacteria in artificial urine also shows that the adhesion is greater for face ([Formula: see text]) than for face (001). The adhesion of bacteria to the examined faces of the struvite crystal, and in particular the increased adhesion of bacteria to the face ([Formula: see text]), may be the first stage of biofilm formation, which may result in a high rate of recurrence of infectious urinary stones after treatment.

Substances Secreted by Lactobacillus spp. from the Urinary Tract Microbiota Play a Protective Role against Proteus mirabilis Infections and Their Complications.

Proteus mirabilis urinary tract infections can lead to serious complications such as development of urinary stones. Lactobacillus spp., belonging to the natural microbiota of the urinary tract, exhibit a number of antagonistic mechanisms against uropathogens, including the secretion of organic acids. In this study, we determined the anti-adhesion, anti-cytotoxicity and anti-crystallization properties of the substances secreted by Lactobacillus. For this purpose, membrane inserts with a pore diameter 0.4 µm were used, which prevent mixing of cultured cells, simultaneously enabling the diffusion of metabolic products. The intensity of crystallization was assessed by measuring the levels of Ca2+, Mg2+ and NH3 and by observing crystals using microscopic methods. The cytotoxicity of the HCV-29 cell line was determined using the LDH and MTT assays, and the impact of lactobacilli on P. mirabilis adhesion to the bladder epithelium was assessed by establishing CFU/mL after cell lysis. It was shown that in the presence of L. gasseri the adhesion of P. mirabilis and the cytotoxicity of the cells decreased. The degree of crystallization was also inhibited in all experimental models. Moreover, it was demonstrated that L. gasseri is characterized by the secretion of a high concentration of L-lactic acid. These results indicate that L-lactic acid secreted by L. gasseri has a significant impact on the crystallization process and pathogenicity of P. mirabilis.

Organic Acids Secreted by Lactobacillus spp. Isolated from Urine and Their Antimicrobial Activity against Uropathogenic Proteus mirabilis.

The natural microbiota of the urinary tract includes Lactobacillus spp., which secrete molecules with antimicrobial properties and have antagonistic activity against many pathogens. This paper focuses on the antibacterial effect of Lactobacillus strains isolated from urine against clinical strains of Proteus mirabilis isolated from kidney stones and from urine with coexisting urolithiasis. The study involved analyzing the main antimicrobial molecules secreted by Lactobacillus. In order to indicate which agent had the strongest antimicrobial effect, the supernatants were made alkaline and treated with catalase and high temperature. Both treated and untreated supernatants were analyzed for their activity. Exposing uropathogens to all untreated cell-free supernatants of Lactobacillus significantly reduced their growth, and it was established that these properties were related to organic acid secretion by these strains. Using LC-MS/MS and spectrophotometric techniques, lactic, citric, and succinic acids were determined qualitatively and quantitatively. The influence of these acids on the P. mirabilis growth and biofilm formation and their influence on membrane permeability were also investigated. The results indicate that organic acids secreted by Lactobacillus strains have a high antibacterial potential and could be used as novel agents in the treatment of urinary tract infections caused by P. mirabilis.

Consumption of soft drinks rich in phosphoric acid versus struvite crystallization from artificial urine.

In recent years, there has been a continuous increase in the incidence of urolithiasis, especially in highly developed countries. Therefore, the question arises which factors specific to these countries may be responsible for the increase in the incidence of this disease. In this article, we try to assess the effect of phosphoric acid, a component of various carbonated drinks, including Coca-Cola, on the nucleation and growth of struvite crystals, which are the main component of infectious urinary stones. The research was carried out in the environment of artificial urine with and without the presence of Proteus mirabilis bacteria. In the latter case, the activity of bacterial urease was simulated by adding an aqueous ammonia solution. The obtained results indicate that phosphoric acid present in artificial urine causes the nucleation of struvite to shift towards a lower pH, which means that struvite nucleates earlier in artificial urine compared to the control test. The amount of struvite formed is the greater the higher the concentration of phosphoric acid. At the same time, as the concentration of phosphoric acid increases, the growing struvite crystals are larger, which is disadvantageous because they are more difficult to remove from the urinary tract along with the urine. For the highest levels of phosphoric acid tested, large dendrites are formed, which are particularly undesirable as they can damage the epithelium of the urinary tract. The effect of phosphoric acid on the nucleation and growth of struvite is explained in base of chemical speciation analysis. This analysis indicates that the MgHCit and MgCit- complexes have the main influence on the nucleation and growth of struvite in artificial urine in the presence of phosphoric acid. It should be keep in mind that all these effects of phosphoric acid are possible when the urinary tract is infected with urease-positive bacteria. In the absence of infection, phosphoric acid will not cause struvite to crystallize.

Potentially Probiotic Lactobacillus Strains Derived from Food Intensify Crystallization Caused by Proteus mirabilis in Urine.

Proteus mirabilis is a common cause of infectious urolithiasis. The first stage in the formation of urinary stones is the crystallization of mineral salts in the urine induced by urease activity of this microorganism. Lactobacillus spp. are an important component of the human microbiota and in large quantities occur in foods. Regardless of their origin, those with probiotic properties are proposed as an alternative to antibiotic therapy in the treatment of urinary tract infections. The aim of the study was to check the effect of selected Lactobacillus plantarum and Lactobacillus brevis strains on crystallization caused by P. mirabilis in an in vitro experiment. It has been confirmed that selected Lactobacillus strains have antibacterial properties and colonize the urinary tract epithelium. During 24-h incubation of bacterial cultures, containing P. mirabilis and individual Lactobacillus strains, in synthetic urine, bacterial viability (CFU/mL), pH, and crystallization were determined. Crystallization was assessed quantitatively and qualitatively using AAS and XRD techniques as well as phase-contrast microscopy. It has been shown that in the presence of selected Lactobacillus strains, the culture pH increases faster, especially after 8 h of incubation, compared with the pure P. mirabilis culture. An increase in pH reduces the viability of P. mirabilis; however, in the presence of some lactobacilli, the uropathogen grows more intensively. The presence of Lactobacillus also affected crystallization by increasing its intensity, and the resulting crystals were larger in size. Tested L. plantarum and L. brevis strains could therefore accelerate the formation of urinary stones and development of infection.

Publisher Correction: Aggregation of poorly crystalline and amorphous components of infectious urinary stones is mediated by bacterial lipopolysaccharide.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Aggregation of poorly crystalline and amorphous components of infectious urinary stones is mediated by bacterial lipopolysaccharide.

Poorly crystalline and amorphous precipitate (PCaAP) is one of the components of the so-called infectious urinary stones, which are the result of the activity of urease-producing microorganisms, mainly from the Proteus species, in particular Proteus mirabilis. The main component of this kind of stones is crystalline struvite (MgNH4PO4∙6H2O). Bacteria can build into the structure of the urinary stone and, in this way, they are one of the components of the urinary stone. From these three components - PCaAP, struvite and Proteus mirabilis - PCaAP exhibits the greatest ability to aggregate. The present study focuses on the aggregation of PCaAP. In particular, an influence of lipopolysaccharide (LPS) isolated from Proteus mirabilis on aggregation of PCaAP is presented. An aggregation of PCaAP is characterized by cross-sectional area of aggregates and zeta potential. The results demonstrate that, in artificial urine, the influence of freely suspended LPS on aggregation of PCaAP depends on the concentrations of LPS. Small concentrations of freely suspended LPS enhance the aggregation of PCaAP compared to the control test. For high concentrations of freely suspended LPS the formation of aggregates of PCaAP is inhibited. LPS, which is not freely suspended, but covers polystyrene latex beads, has no such properties. The investigations provide evidence for the importance of biological regulation in the PCaAP aggregation process.

Influence of various uropathogens on crystallization of urine mineral components caused by Proteus mirabilis.

Infectious urolithiasis is a consequence of long-standing urinary tract infections with urease-positive bacteria, especially Proteus spp. However, because of the often mixed nature of urinary tract infections, in the case of urinary stones formation, several species of bacteria may be involved in the process. The purpose of the study was to determine the impact of the bacterial species: Escherichia coli, Klebsiella pneumoniae, Providencia stuartii, Pseudomonas aeruginosa and Staphylococcus aureus on the crystallization caused by Proteus mirabilis. The studies were conducted in synthetic urine with the addition of P. mirabilis and a representative of another species. During the experiments the viability of bacteria, pH, presence and morphology of crystals, and the intensity of crystallization were assessed. Crystallization of calcium and magnesium phosphates occurred in all investigated configurations. However, there were differences observed in the course and intensity of crystallization between the mixed culture and the P. mirabilis culture. Although most intense crystallization took place in the pure culture of P. mirabilis it was also demonstrated that the presence of other uropathogens increased the survival of P. mirabilis. This synergistic effect could be responsible for the persistence and recurrence of urolithiasis in the urinary tract.

Development of a molecular serotyping scheme and a multiplexed luminex-based array for Providencia.

Providencia is an opportunistic human pathogen that belongs to the Enterobacteriaceae family. The bacterial cell surface O-antigen is one of the most structurally variable cell constituents and serves as a basis for serotyping gram-negative bacteria. In this work, the genomes of 12 Providencia strains were sequenced, and genes driving O-antigen biosynthesis were analyzed. The O-antigen-synthesizing genes of Providencia are located in the O-antigen gene cluster (OGC) between the cpxA and yibK genes. The gene functions predicted in silico agreed with the known O-antigen structures. All clusters were found to contain both wzx and wzy and exhibit a high degree of heterogeneity. Based on the sero-specific genes, we developed a molecular serotyping system to detect 23 serotypes (from the present and previous studies) for the first time. Five Proteus strains, five Morganella strains, five uropathogenic Escherichia coli (UPEC) strains and 32 Providencia strains with other serotypes were used to assess the specificity of our multiplexed Luminex-based array. Five serogroups (O3, O8, O19, O38 and O52 strains) were used to determine the sensitivity of the suspension array. The detection sensitivity was 0.1 ng genomic DNA, 103 CFU/ml in pure culture, or 104 CFU/ml in mock urine specimens. Furthermore, 29 publicly available Providencia genomes (which have not been serotyped) were analyzed, and 23 novel putative OGC types were identified. In total, we identified 35 new OGCs and developed a molecular serotyping system based on the sero-specific genes. The established classification system can support promising applications in basic research, clinical diagnosis, and epidemiological surveillance.

Genetic diversity of the O antigens of Proteus species and the development of a suspension array for molecular serotyping.

Proteus species are well-known opportunistic pathogens frequently associated with skin wound and urinary tract infections in humans and animals. O antigen diversity is important for bacteria to adapt to different hosts and environments, and has been used to identify serotypes of Proteus isolates. At present, 80 Proteus O-serotypes have been reported. Although the O antigen structures of most Proteus serotypes have been identified, the genetic features of these O antigens have not been well characterized. The O antigen gene clusters of Proteus species are located between the cpxA and secB genes. In this study, we identified 55 O antigen gene clusters of different Proteus serotypes. All clusters contain both the wzx and wzy genes and exhibit a high degree of heterogeneity. Potential functions of O antigen-related genes were proposed based on their similarity to genes in available databases. The O antigen gene clusters and structures were compared, and a number of glycosyltransferases were assigned to glycosidic linkages. In addition, an O serotype-specific suspension array was developed for detecting 31 Proteus serotypes frequently isolated from clinical specimens. To our knowledge, this is the first comprehensive report to describe the genetic features of Proteus O antigens and to develop a molecular technique to identify different Proteus serotypes.

Binding of CXCL8/IL-8 to Mycobacterium tuberculosis Modulates the Innate Immune Response.

Interleukin-8 (IL-8) has been implicated in the pathogenesis of several human respiratory diseases, including tuberculosis (TB). Importantly and in direct relevance to the objectives of this report quite a few findings suggest that the presence of IL-8 may be beneficial for the host. IL-8 may aid with mounting an adequate response during infection with Mycobacterium tuberculosis (M. tb); however, the underlying mechanism remains largely unknown. The major goal of our study was to investigate the contribution of IL-8 to the inflammatory processes that are typically elicited in patients with TB. We have shown for the first time that IL-8 can directly bind to tubercle bacilli. We have also demonstrated that association of IL-8 with M. tb molecules leads to the augmentation of the ability of leukocytes (neutrophils and macrophages) to phagocyte and kill these bacilli. In addition, we have shown that significant amount of IL-8 present in the blood of TB patients associates with erythrocytes. Finally, we have noted that IL-8 is the major chemokine responsible for recruiting T lymphocytes (CD3(+), CD4(+), and CD8(+) T cells). In summary, our data suggest that the association of IL-8 with M. tb molecules may modify and possibly enhance the innate immune response in patients with TB.

Various intensity of Proteus mirabilis-induced crystallization resulting from the changes in the mineral composition of urine.

Infectious urolithiasis is a result of recurrent and chronic urinary tract infections caused by urease-positive bacteria, especially Proteus mirabilis. The main role in the development of this kind of stones is played by bacterial factors such as urease and extracellular polysaccharides, but urinary tract environment also contributes to this process. We used an in vitro model to establish how the changes in the basic minerals concentrations affect the intensity of crystallization which occurs in urine. In each experiment crystallization was induced by an addition of P. mirabilis to artificial urine with a precisely defined chemical composition. Crystallization intensity was determined using the spectrophotometric microdilution method and the chemical composition of formed crystals was established by atomic absorption spectroscopy and colorimetric methods. Increasing the concentration of all crystals forming ions such as Mg(2+), Ca(2+) and phosphate strongly intensified the process of crystallization, whereas reducing the amount of these components below the proper physiological concentration did not affect its intensity. The inhibitory influence of citrate on calcium and magnesium phosphate crystallization and competitive actions of calcium and oxalate ions on struvite crystals formation were not confirmed. In the case of infectious stones the chemical composition of urine plays an important role, which creates a necessity to support the treatment by developing a model of proper diet.

In vitro studies of epithelium-associated crystallization caused by uropathogens during urinary calculi development.

Infectious urinary stones account for about 10% of all urinary stones. In 50% of cases urolithiasis is a recurrent illness, which can lead to the loss of a kidney if not properly treated. One of the reasons for recurrence of the disease may be the ability of bacteria to invade urothelial cells, persist in the host cells and serve as potential reservoirs for infection. Various uropathogens are associated with the formation of bacteria-induced urinary stones but Proteus mirabilis is the most commonly isolated (70%). An in vitro model was used in this study to analyze intracellular growth and crystallization in the presence of P. mirabilis, Klebsiella pneumoniae and Escherichia coli. Human ureter (Hu 609) and bladder (HCV 29) epithelial cell lines were infected with bacteria and incubated (3-72 h) in the presence of synthetic urine and amikacin to prevent extracellular bacterial growth. During the incubation the number of bacteria (CFU/ml) inside epithelial cells and the intensity of crystallization were established. Crystallization was determined as an amount of a calcium radioisotope. The chosen strains of uropathogens were able to invade both types of epithelial cells but the Hu 609 cells were invaded to a higher extent. However, crystallization occurred only in the presence of P. mirabilis strains which were invasive and urease-positive. The highest intensity of cell-associated crystallization was observed when the number of bacteria within the urothelium remained stable during the time of incubation. These results show that P. mirabilis has an ability to form crystals inside the host cells. Under these conditions bacteria are protected from antibiotic killing, which leads to persistent and recurrent infections. We also suspect that this phenomenon may be an important stage of kidney stones formation.

Inhibition of crystallization caused by Proteus mirabilis during the development of infectious urolithiasis by various phenolic substances.

Infectious urolithiasis is a consequence of persistent urinary tract infections caused by urease producing bacteria e.g. Proteus mirabilis. These stones are composed of struvite and carbonate apatite. Their rapid growth and high recurrence indicate that so far appropriate methods of treatment have not been found. In the present study, the inhibitory effect of phenolic compounds was investigated in vitro against formation of struvite/apatite crystals. The impact of these substances with different chemical structures on crystallization caused by clinical isolates of P. mirabilis was tested spectrophotometrically using a microdilution method. Among the 11 tested compounds resveratrol, epigallocatechin gallate, peralgonidin, vanillic and coffee acids at the concentrations 250-1000 µg/ml inhibited P. mirabilis urease activity and crystallization. However, only vanillic acid had such an effect on all tested strains of P. mirabilis. Therefore, using an in vitro model, bacterial growth, crystallization, urease activity and pH were examined for 24h in synthetic urine with vanillic acid. Effect of vanillic acid was compared with that of other known struvite/apatite crystallization inhibitors (acetohydroxamic acid, pyrophosphate) and it was shown that vanillic acid strongly inhibited bacterial growth and the formation of crystals. It can be assumed that this compound, after further studies, can be used in the treatment or prophylaxis of infectious urolithiasis.

In vitro studies on the role of glycosaminoglycans in crystallization intensity during infectious urinary stones formation.

Proteus mirabilis cause urinary tract infections which are recurrent and can lead to formation of urinary calculi. Both bacterial and the host factors are involved in the development of urolithiasis. To determine the impact of glycosaminoglycans (GAGs) in the formation of P. mirabilis-induced urinary stones, we investigated the in vitro crystallization, aggregation and adhesion of crystals in the presence of GAGs naturally appearing in urine. Crystallization experiments were performed in synthetic urine infected with P. mirabilis in the presence of: hyaluronic acid (HA), heparan sulfate (HS), chondroitin sulfate A, B and C (ChSA, ChSB, ChSC). The intensity of crystallization and aggregation were established by counting particles and phase-contrast microscopy. To analyze the adhesion of crystals, we used normal urothelium and (45)Ca isotope-labeled crystals. In the presence of ChSC, both the size of the crystals formed and their number were higher compared with the control. GAGs increased crystals adhesion to the cells, but only for ChSA this effect was significant. Chondroitin sulfates, which accelerate the first stages of infection-induced stones formation, may play an important role in the pathogenesis of infectious urolithiasis.

Analysis of Proteus mirabilis distribution in multi-species biofilms on urinary catheters and determination of bacteria resistance to antimicrobial agents.

The objectives of the investigation presented in this paper were: to examine the frequency of P. mirabilis isolation from catheters and assess the complexity of multi-species biofilms which these bacteria form, as well as to determine the vulnerability of planktonic and sessile P. mirabilis populations to popular antibiotics and compare it to the susceptibility of other Gram-negative bacteria isolated as associated flora from multi-species biofilm. 88 urological catheters, collected from long-term catheterized patients were examined. Uropathogens were recovered from the catheter surface by sonication, and identified on standard diagnostic media. The broth-microdilution method and the MBEC High-throughput Screening assay were used to determine the bacterial resistance to antibiotics. 279 microorganisms were isolated from 88 urinary catheter biofilms. The Enterobacteriaceae family were the most frequently detected bacteria (53.2% of isolates), whereas Proteus spp. isolation accounted for 17.9%, which placed these bacilli on the third position in the Enterobacteraceae family. Among all the tested drugs, amikacin and cephalosporins (ceftriaxone, cefotaxime and cefaclor) exhibited the highest activity against P. mirabilis planktonic cells, 86% and 73% of strains were susceptible to these antibiotics, respectively. 100% of P. mirabilis sessile forms were resistant to cefepime, ciprofloxacin, gatifloxacin, and norfloxacin. Amikacin and ceftriaxone affected only 5% of sessile forms. The planktonic cells of the other studied uropathogens were mostly vulnerable to the all tested drugs (exception P. aeruginosa strains), the most effective of which occurred to be amikacin and cefepime. Obtained MBECs values were 2-512-fold higher than MICs assessed for planktonic forms.

[Oxalobacter formigenes--characteristics and role in development of calcium oxalate urolithiasis]. / Oxalobacter formigenes--charakterystyka i rola w rozwoju kamicy szczawianowo-wapniowej.

Microorganisms are one of the important factors for urinary calculi formation. While urease-positive bacteria and nanobacteria contribute to stone formation, Oxalobacter formigenes rods play a protective role against the development of urolithiasis. Proteus mirabilis alkaline environment of the urinary tract and cause crystallization mainly of struvite (magnesium ammonium phosphate). However, nanobacteria, due to the possibility of apatite deposition on the surface of their cells, have long been considered as an etiological factor of urinary calculi consisting of calcium phosphates. O. formigenes is an anaerobe using oxalate as the main source of carbon and energy and occurs as natural gastrointestinal microflora of humans and animals. These bacteria control the amount of oxalate excretion degrading oxalates and regulating their transport by intestinal epithelium. Lower colonization of the human colon by O. formigenes can cause increased oxalate excretion and lead to the development of oxalate urolithiasis. Due to the positive influence of O. formigenes, there is ongoing research into the use of this microorganism as a probiotic in the prophylaxis or treatment of hyperoxaluria, both secondary and primary. The results of these studies are very promising, but they still require continuation. Future studies focus on the exact characteristics of O. formigenes including their metabolism and the development of methods for applying as a therapeutic agent the bacteria or their enzymes degrading the oxalate.

Unique surface and internal structure of struvite crystals formed by Proteus mirabilis.

Crystallization of struvite from artificial urine in the presence of Proteus mirabilis microorganisms depends strongly on pH value. At small value of pH, struvite yields crystals of coffin-like habit with very specific structure. The analysis using scanning electron microscopy shows that the crystals possess well-defined faces, but higher magnifications show very specific structuration as if the crystals were built from small three-dimensional subunits. The possible role of microorganisms in the formation of such a structuration is analyzed. At higher pH value, the crystals exhibit dendritic growth with main trunk and branches. Although the formation mechanism of the specific structuration as well as dendritic structures is unknown, the nature of forces for such an alignment is analyzed. The revealed porous internal structure of struvite is also analyzed. The investigations provide evidence for the importance of biological regulation in crystallization process.