Esquema general que debe llevar la metodología

1. Determinación in vitro las concentraciones y el tiempo de contacto de los desinfectantes

1.1. Concentración y tiempo de contacto del peróxido de hidrógeno

1.2. Concentración y el tiempo de contacto del peróxido de hidrógeno

2. Muestreo:

2.1 Obtención de muestra de agua de la jeringa triple

2.2 Recolección de muestra de agua de la turbina

2.3 Recogida de muestra de agua del tanque reservorio

2.4 Traslado de la muestras al laboratorio

3. Análisis Microbiológico de las muestras

3.1 Cuantificación de bacterias aerobia mesófilas

3.2 Determinación de contaje de Mohos y levaduras

3.3 Detección de colonias presuntivas Samonella sp.

4. Análisis de los resultados

3.3. Detección de colonias presuntivas de Salmonella sp.

3.3.1 Aislamiento de colonias presuntivas Salmonella:

*3.3.1.1 Enriquecimiento de la muestra,

**3.3.1.2 Crecimiento selectivo en agar Bilis Verde Brillante, selección de colonias presuntivas
de Salmonella. Sacarlo de Aquí (lo saqué del material enviado):
 *Enrichment: . Tetrathionate broth, incubated at 35°C, inhibits coliforms and Gram-positive
bacteria, permitting selective enrichment of most Salmonella species, including S. typhi. It has
been reported that tetrathionate broth is more selective for Salmonella than selenite-based
media when incubated for 48 h at 43°C. While this formulation is highly selective, it is unable
to inhibit Proteus mirabilis, which shows optimum growth. Growth of Proteus and Citrobacter
can be inhibited with addition of brilliant green (see Section 9260B.3a). Incubation at 43°C and
addition of brilliant green also will inhibit some species of Salmonella, including S. typhi.

** Selective Growth:

a. Brilliant green agar: Typical well-isolated Salmonella colonies grown on this medium are
pinkish white with a red background. S. typhi and a few other species of Salmonella grow
poorly because of the brilliant green dye content. Lactose-fermenters not subject to growth
suppression will form greenish colonies or may produce other colorations. Occasionally, slow
lactose-fermenters (Proteus, Citrobacter, and Pseudomonas) will produce colonies resembling
those of a pathogen. Suppress spreading effect of pseudomonads by increasing agar
concentration to 2%. In some instances, Proteus has been observed to ‘‘swarm’’; reduce this
tendency by using agar plates dried to remove surface moisture. If suspect Salmonella colonies
are not observed after 24 h incubation, reincubate for an additional 24 h to permit slow-
growing or partially inhibited organisms to develop visible colonies. If typical colonies are not
observed or if the streak plate is crowded, isolate in pure culture a few colonies for
biochemical characterization. Non-lactose-fermenting colonies in close proximity to lactose-
fermenting colonies may be masked.

Esto se pone al final del procedimiento :Las colonias presuntivas de Salmonella se guardaran
en agar Tripticasa de soya inclinado, a una temperatura de 10°C para su posterior
identificación.

Detección de Mohos y Levaduras

a. Fungi in potable water: Fungi have been found in potable water2-7 and on the inner
surface of distribution system pipes.8 Either they survive water treatment or they enter the
system after treatment and remain viable. Tuberculate macroconidia of Histoplasma
capsulatum9
can pass through a 0.75-m rapid sand filter. Plain sedimentation or alum flocculation and
settling
removed 80 to 99% of the spores. If these relatively large (8- to 14-μm) globose,
tuberculate
macroconidia pass through treatment, it is not surprising that other fungi, typically with
smaller
spores, are found in treated water.
Containers: Collect samples as directed in Section 9060A. Alternatively, use cylindrical
plastic vials with snap-on caps. These vials usually are sterile as received. Transport them
in an
upright position to minimize the chance of leakage and discard after use.
b. Storage: Hold samples for not more than 24 h. If analysis is not begun promptly after
sample collection, refrigerate.

Media: El medio a utilizar es el Agar Saboraud

Procedure
As many as 40 samples can be analyzed simultaneously by a single analyst by the
following
procedure; however, 20 samples represents the optimum number.
b. Plating:
Add 1 mL of appropriate sample and mix thoroughly by
tilting and rotating dish (see plating procedure under heterotrophic plate count, Section
9215).
add agar medium at 43 to 45°C. Solidify agar as rapidly as possible. (In arid areas use more
medium
to prevent dehydration during incubation.)
c. Incubation: Stack plates but do not invert. Incubate at room conditions of temperature
(20
to 24°C) and lighting but avoid direct sunlight. Examine and count plates after 3, 5, and 7 d.
d. Counting and inventory: The fungus plate count will provide the basis for rough
quantitative comparisons among samples; the inventory will give relative importance of at
least
the more readily identifiable species or genera.
In preparing plates, use sample portions that will give about 50 to 60 colonies on a plate.
Determine this volume by trial and error. When first examining a new habitat plate at least
two
sample dilutions. Estimates of up to 300 colonies may be made, but discard more crowded
plates. The medium containing rose bengal tends to produce discrete colonies and
permits