Evaluation of the impact of technical and technological adjustments to the water treatment plant on the quality of treated water

The subject of this bachelor thesis is the evaluation of sequential technical and technological changes of water collection and modification at a specific water treatment facility – the water treatment facility in Ivančice. The first part of the thesis explains the basic terminology such as technological processes of the water treatment plant, types of water treatment facilities, the range of different types of water based on its quality and technologies that can be found at the water treatment plant in Ivančice. The next two chapters describe the water treatment process before and after intensification. The primary focus is on raw water collection, the quality of both raw and treated water, and the water treatment technology. The last part of the paper offers multiple comparisons of the original and renovated facility and recommendations for improving the operations of the renovated water treatment facility. The final evaluation was carried out based on the information collected mainly by visiting the facility in person, and other sources

ASC is involved in the <i>S. aureus</i>-induced IL-1alpha and IL-1beta release in keratinocytes.

<p>Primary keratinocytes were left unstimulated (- SA) or were stimulated with <i>S. aureus</i> (+ SA) for a total of six hours. To investigate the influence of ASC on the IL-1alpha and IL-1beta expression the cells were transfected with an ASC specific siRNA or a non-silencing control siRNA. Gene expression of IL-1alpha (A) and IL-1beta (B) was measured by real-time PCR, protein secretion of IL-1alpha (C) and IL-1beta (D) was analysed by ELISA. Data are means ± SEM (*p< 0.05, n.s. = not significant, Student`s <i>t</i>-test, n = 6).</p

Infection of primary keratinocytes with <i>S. aureus</i> induces expression of IL-1alpha and IL-1beta.

<p>Human primary keratinocytes were left unstimulated (- SA) or were stimulated with <i>S. aureus</i> (+ SA) for a total of six hours. Gene expression of IL-1alpha (A) and IL-1beta (B) was analysed by real-time PCR. Protein secretion of IL-1alpha (C) and IL-1beta (D) was measured by ELISA. Data are means ± SEM (**p< 0.01, Student`s <i>t</i>-test, n = 3).</p

<i>Trichophyton rubrum</i> together with the cytokine combination IFN-γ/IL-17 synergistically induce RNase 7 and hBD-3 expression in keratinocytes.

<p>(A) Primary keratinocytes were incubated with 1×10⁷/ml conidia of <i>T. rubrum</i> together with IFN-γ/IL-17A (each 20 ng/ml) for 24 h. Gene expression of RNase 7 (a) and hBD-3 (c) was analyzed by real-time PCR. Release of RNase 7 was determined by ELISA (b). Data are means ± SD (n = 3; *p<0.05, **p<0.01, Student's <i>t</i> test).</p

Infection of keratinocytes with <i>Trichophyton rubrum</i> leads to increased RNase 7 secretion.

<p>Primary keratinocytes were exposed to 1×10⁶/ml and 1×10⁷/ml conidia of <i>T. rubrum</i> for 24 h. Secretion of RNase 7 was determined by analysis of the presence of RNase 7 in the cell supernatants using ELISA. Data are means ± SD (n = 3; *p<0.05, **p<0.01, Student's <i>t</i> test).</p

Epidermal keratinocytes express RNase 7 <i>in vivo</i>.

<p>(A & B) Immunostaining of RNase 7 expression in human normal skin using affinity-purified RNase 7 antibodies. Strong RNase 7 immunoreactivity was detected in the epidermis with highest activity in the uppermost epidermal layers. Sebaecous glands and hair follicles also stained positively. (A) 10×magnification, (B) 20 X magnification of the indicated area of panel A. (C) Negative control using preimmune serum. (E: Epidermis, SC: Stratum Corneum, HF: Hair Follicle, I: Infundibulum, SG: Sebaceous Gland, DP: Dermal Papilla, ORS: Outer Root Sheath, IRS: Inner Root Sheath)</p

RNase 7 contributes to the killing activity of human stratum corneum extracts against <i>E. faecium</i>.

<p>(A) RNase 7 (12.5 µg⋅ml⁻¹) was tested in a microdilution assay against <i>E. faecium</i> (ATCC 6057) alone (R7) or in the presence of 10 mg⋅ml⁻¹ RNase 7 antibodies (R7+R7-Ab). Application of the RNase 7 antibodies completely blocked the <i>E. faecium</i>-killing activity of RNase 7. As a control, RNase 7 was incubated with irrelevant goat antibodies (R7+irr.Ab). Both, RNase 7 antibodies (R7-Ab) alone as well as irrelevant antibodies (irr. Ab) alone did not influence the growth of <i>E. faecium</i>. (B) The killing activity of skin-extracts derived from stratum corneum against <i>E. faecium</i> was tested. 2 h incubation of <i>E. faecium</i> with stratum corneum extract revealed high killing activity of the extract against <i>E. faecium</i> (s.c.). In contrast, the application of RNase 7-blocking antibodies to the stratum corneum extract significantly reduced the killing activity of the extract (s.c.+ R7-specific Ab; p<0.01, Student's <i>t</i>-test). Incubation of the stratum corneum extract with the irrelevant antibodies did not affect the killing activity of the extracts (s.c.+ irrelevant Ab). Data show means±S.D. of triplicate samples. A representative result of three independent experiments is shown.</p

An antibody towards the epidermal growth factor receptor (EGFR) inhibited the <i>Trichophyton rubrum</i>-induced RNase 7 and hBD-3 expression in keratinocytes.

<p>Primary keratinocytes were pre-incubated with the EGFR-antibody cetuximab (20 µg/ml) followed by treatment for 24 h with 1×10⁷/ml conidia of <i>T. rubrum</i> in the presence of cetuximab (20 µg/ml). Gene expression of RNase 7 (a) and hBD-3 (c) was analyzed by real-time PCR. Release of RNase 7 was determined by ELISA (b). Data are means ± SD (n = 3; *p<0.05, **p<0.01, Student's <i>t</i> test).</p

HBD-3 restricts the growth of <i>Trichophyton rubrum</i>.

<p>45 µl of germinating conidia of <i>T. rubrum</i> were incubated in a 96-well plate for 3 h at 34°C in 10 mM sodium phosphate buffer (pH 7.2) together with 5 µl of hBD-3 (solved in 0.01% acetic acid) to reach an end concentration of 50 µg/ml hBD-3 (a), 25 µg/ml hBD-3 (b), 12.5 µg/ml hBD-3 (c) and 6.25 µg/ml hBD-3 (d). Incubation without hBD-3 (only 5 µl 0.01% acetic acid) served as a growth control and treatment with the antifungal drug fluconazole (100 µg/ml end concentration) served as a positive control. After 3 h incubation 150 µl Sabouraud's glucose broth containing 100 units/ml penicillin and 100 µg/ml streptomycin was added to each well and growth was monitored over time by measuring the optical density at 620 nm. Data are means ± SD of one representative experiment of two, each performed in triplicate samples (*p<0.05, **p<0.01 compared to growth control, Student's <i>t</i> test).</p

IFN-γ/IL-17 enhance the antimicrobial activity of primary keratinocytes.

<p>Primary keratinocytes were stimulated with IL-17A (40 ng/ml), IFN-γ (40 ng/ml) or IFN-γ/IL-17A (each 20 ng/ml) for 24 h. Cell lysates were incubated with <i>Staphylococcus aureus</i> and bacteria growth was determined by measuring optical density at 600 nm (OD₆₀₀). Data are derived from three experiments each done in triplicates and statistical differences were analyzed using one-way ANOVA with Tukey’s test (*p<0.05, ns = not significant).</p