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This study was conducted on a private small cattle breeders farms located
in Beni-Suef (coordinates: 29°04’N31°05’E) and Fayoum
(coordinates: 29°308374?N–30°844105?E) province, Egypt throughout a period from May to December 2017.
Representative water samples were collected from both main source and water
troughs of both supplies (tap and hand pumps water) at small cattle breeders (n=
15) in the study areas.

Study design

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The protocol of this study divided into two steps to investigate and
improve the hygienic quality of drinking water at small cattle breeders. The
first one aimed to evaluate the biocidal activity of calcium hypochlorite as a
common disinfectant used for drinking water and detect the extent of its
efficacy against waterborne pathogenic bacteria then using nanocomposite
contains silver nanoparticles loaded Ca(OCl)2 for trying to
enhuncement the disinfectant performance. Stratified water samples were
collected from small cattle breeders for isolation and identification of waterborne
pathogenic bacteria using biochemical test and a molecular assay.  The sensitivity of 50 bacterial strains were
evaluated to tested disinfectant, AgNPs and AgNPs/Ca(OCl)2 nanocomposite
using broth macro dilution method. The second is a field trial for the sake of
evaluating the efficacy of bactericidal filter paper against indicators
coliforms bacteria. Water samples were bacteriologically examined prior and
post application of treated filter paper for total viable bacteria, total
coliform and fecal coliform counts. All collected data were recorded and
statistically analyzed.

Water sampling

Under aseptic condition, a total of 100 water samples were collected
from both main source and water troughs used for cattle drinking at small
cattle breeders in the investigated area. The collected sample were obtained
from two water supplies (tap and hand pump water) in a 250 mL sterilized
tightly capped bottle and properly labeled. The outlet of both tap and hand pumps were thorough disinfected using
ethyl alcohol 70 %, allowed water flow, and then water samples were taken.
Samples were transferred immediately to the lab in an ice box for further
microbiological examination according to standard water guidelines (APHA
2012).

Isolation and identification of waterborne pathogenic bacteria

Samples were cultured on plate count agar (………….) for detection and
enumeration of total viable count (TVC), using pour plate method as described
by APHA (2012), where the colonies developed after incubating at 37°C
for 24 h. Total coliform (TCC) was enumerated on M-Endo LES agar (Difco,
Sparks, MD) while fecal coliform (FCC) was counted on M-FC agar (EM Science,
Gibbstown, NJ) using conventional membrane filtration (MF) technique as
described by APHA (2012). Each of E. coli and Klebsiella
spp. were isolated on MacConkey agar (Oxoid; CM 0115) and Eosine Methylen Blue
(EMB) agar (Oxoid; CM 69) plates; S. aureus was isolated on Baird Parker
agar base (Oxoid Ltd, Hampshire, England; CM 0275) with egg yolk supplement. Furthermore,
all isolated bacterial colonies are sub-cultured on nutrient agar for
purification. Enteric bacteria was identified on the base of their colony,
morphology, and using API 20E (Biomerieux, Crappone France). S. aureus
strains was purified phenotypically by the tube coagulase test and StaphID32API
systems (API System, BioMe’rieux, Paris, France).

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