Current procedures for the detection and identification of bacterial infections are laborious, time-consuming, and require a high workload and well-equipped laboratories. Therefore the work presented herein developed a simple, fast, and low cost method for bacterial detection based on hydroxyapatite nanoparticles with a nutritive mixture and the fluorogenic substrate. Calcium phosphate ceramic nanoparticles were characterized and integrated with a nutritive mixture for the early detection of bacteria by visual as well as fluorescence spectroscopy techniques. The composite was obtained by combining calcium phosphate nanoparticles (Ca:P ratio, 1.33:1) with a nutritive mixture of protein hydrolysates and carbon sources, which promote fast bacterial multiplication, and the fluorogenic substrate 4-methylumbellipheryl-β-D-glucuronide (MUG). The composite had an average particle size of 173.2 nm and did not show antibacterial activity against Gram-negative or Gram-positive bacteria. After an Escherichia coli suspension was in contact with the composite for 60-90 min, fluorescence detected under UV light or by fluorescence OPEN ACCESSMolecules 2014, 19 13949 spectrophotometer indicated the presence of bacteria. Intense fluorescence was observed after incubation for a maximum of 90 min. Thus, this calcium phosphate nanocomposite system may be useful as a model for the development of other nanoparticle composites for detection of early bacterial adhesion.

Nano- or microhydroxyapatites with microbiological properties are being used to detect pathogens in clinical samples and industrial environments. In this study, the calcium phosphates coral-hydroxyapatite and biphasic calcium phosphate were characterized physicochemically using x-ray diffraction, thermogravimetric, and differential thermal analysis. The morphology, texture, and chemical composition of the ceramics were also investigated using scanning electron microscopy with energy dispersive spectroscopy. The biocompatibility of the ceramics was evaluated using Escherichia coli and Enterococcus faecalis. Microorganisms were detected by incorporating the enzyme markers 4-metilumbelliferil-β-d-glucoside and 4-metilumbelliferil-β-d-glucuronide in the ceramic powders and evaluating fluorescence. The characterization of the ceramics revealed typical characteristics, such as crystallinity, thermal stability, and chemical composition, consistent with other calcium phosphates. The calcium phosphates coral-hydroxyapatite and biphasic calcium phosphate ceramics differed from one another in morphology, structural topography, particle size distribution, and the capacity to absorb water. These properties can influence the rates of microbiological responses and bacterial detection. Although both materials are suitable for use as structural supports in microbial diagnostic systems, BCP was more efficient and detected E. coli and E. faecalis more rapidly than CHA.

A new chromogenic method for the detection and enumeration of enterococci in water was evaluated using water samples from different sources, specifically, industrial, potable and recreational waters, and compared with the standard methodology for water quality ISO 7899-2. Several statistical parameters, such as relative accuracy, selectivity, efficiency, limit of detection, limit of quantification, inclusivity and exclusivity were determined according to the main criteria provided in ISO 16140 and ISO 17994. An orthogonal regression analysis demonstrated that the developed alternative method performs similar to ISO 7899. The chromogenic medium allowed easy recognition of enterococci by the appearance of pinkcolored colonies. Importantly, the method can be executed in only 24 h, showing a similar selectivity (-0.05), a lower limit of quantification (4.08 cfu/100 ml), and a higher efficiency (98%) than the reference method (-0.04; 5.50 cfu/100 ml and 97%, respectively). The results obtained support the use of the alternative method for the detection and enumeration of enterococci to assess water quality from several sources.

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