Monitoring of Fecal Pollution in Fresh Water and Soil of Abbis Area , East of Alexandria , Egypt

Great concern is currently assigned to monitor the environmental conditions at Abbis and its satellite villages, east of Alexandria City, Egypt that suffer severe fecal pollution arising from the lack of sanitary sewerage system. The main objective of the present study was to evaluate the bacteriological quality of water and soil at Abbis area (Villages no. 7 and 8) and some physicochemical characteristics of the water bodies. Results revealed that pH, salinity, dissolved oxygen, biodegradable organic matter and chemical oxygen demand exceeded their permissible limits. The total viable bacterial count ranged between 1.0 x10 and 6.2 x10 CFU/ml in water samples and from 7.1x10 to 8.0 x10 CFU/g in soil samples. Total coliforms in the water of the less polluted sites recorded <300 cell/100 ml and in the highly polluted sites recorded >1800 cell/100 ml. However, all soil samples were heavily contaminated with total coliforms (>1800 cell/100 g). Fecal coliforms in the water samples ranged from 5 to 350 cell/100 ml. For soil, half of soil samples showed very high density of fecal coliforms (>1800 cell/100 g) and the rest showed fecal coliforms in the range of 35-1600 cell/100 g. Some correlations were detected among the tested water parameters at the study area and most of the sampling sites were similar. According to the European and Egyptian current standards, the bacteriological analyses confirmed the results of the physicochemical characterization where most of the selected sites were highly polluted.


INTRODUCTION
The most challenging problem facing the world now and in the near future may be the availability of clean and safe fresh water.It was reported that over two billion people in the world did not have access to safe water supply and sanitation services. (1)Water in sufficient quantity and of adequate quality is necessary for the well-being of all living organisms.The importance and intensive use of fresh Bull High Inst Public Health Vol.39 No.3 [2009]   water makes it a vulnerable and increasingly limited resource.A wide range of human activities may lead to environmental deterioration of surface water, either directly or indirectly. (2)e way domestic pollution affects water quality heavily depends on the way of disposal of this pollution.Approximately, 65% of Egypt's population is connected to drinking water supply, and only 24% to sewage services and most drains are heavily polluted with sewage. (3)Rural areas sewerage and sewage systems are virtually non-existent in rural areas.There is a clear need for arrangements to reduce the ongoing discharge of completely untreated sanitary waste water into the river. (3)though high numbers of water-borne diseases are reported, it is believed that many more people suffer from diseases related to other forms of water pollution. (3)otection of the public and environmental health requires safe, clean water, which means that it must be free of pathogenic bacteria .(4)Heterotrophic bacteria play an important role in the structure and function of the microbial food web in relation to environmental conditions. (5,6)ong the pathogens disseminated in water sources, enteric pathogens are the most frequently encountered ones.As a consequence, sources of fecal pollution in waters devoted to human activity must be strictly controlled.Survival rates of fecal bacteria can vary from a few minutes to many days depending upon the environmental conditions. (14,15)r Physical factors including solar radiation.(16,17) , temperature, (15,18) salinity, (15,19) and dissolved oxygen. (15) Abis villages no. 7 and 8 and their salite villages are important from the environmental point of view since they are suffering a non-sanitary sewerage conditions that does not affect water and soil only, but also the whole community.
The lack of a sanitary drainage system means that residents dispose of their sewage in the nearby watercourses.Moreover, some physicochemical l characteristics of the water in the study area were examined.

Sampling Sites and Sampling
Fresh water and soil samples were collected from Abbis agricultural villages no.7 and 8 located in Beheira Governorate.The area at the south of Lake

Bacteriological Examination
The freshly aseptically collected 30 soil samples (100 g each) were shaken in 100 agar plates and incubated at 37°C for 24 h.
Microscopic examination was carried out to ensure gram-negative, non-spore forming rods.The MPN-index per 100 ml was determined using the statistical tables.

Statistical Analysis
Pair-wise correlation coefficients between various physical, chemical and bacteriological parameters were calculated.
Positive and negative associations were

Physicochemical Characteristics
The values of physicochemical characteristics in 1. Bacteriological examination of the water samples at the selected sites showed that they were highly polluted.This is clearly shown by the numbers of the TVC obtained in the water samples from the selected canals, which ranged between 1.0x10 4 and 6.2x10 5 CFU/ml.
3. FC in the water samples ranged between a minimum of 5 and a maximum of 350 cell/100 ml (Table 2 and Figure 3).

Bacteriological Status of Soil at Abbis
According to the results given in Table 3, soil at the selected sites showed huge number of bacterial populations with TVC density that ranged between 7.1x 10 6 and 8.0 x 10 9 CFU/g.
Except for site 30, all sites were heavily contaminated with total coliform bacteria (>1800 cell/100g), as shown in Figure 4.This was confirmed by the results of fecal coliform , where 15 sites out of the 30 showed very high density of FC (>1800 cell/100g) indicating recent fecal pollution while other sites recorded FC that ranged between 35-1600 cell/100g (Figure 5).Analysis for the data of the 30 sampling sites using the similarity coefficient test is shown in the dendogram (Figure 6).Environmental Affair Agency, (1994), (25) revealed the degree of pollution in the study area.The hydrographical parameters indicated that pH values were slightly toward the alkaline side and lie at the optimum level for most microorganisms. (26)ey were also within the normal and suitable range of irrigation water (MPL 6.5-

8.4) at the different sites. Salinity exceeded
Bull High Inst Public Health Vol.39 No.3 [2009]   the maximum permissible limit (MPL<0.36g/l) at most sites with differences among them.Water salinity is considered as a sensitive parameter for measuring the rate of outfall discharge and subsequently it reflects the degree of pollution in aquatic environments.Salinity can affect viability of the fecal bacteria and their survival. (15,19)ssolved oxygen contents of water samples were above the MPL (5 mg/l) at all sites (Table 1).Dissolved oxygen is an important parameter for the identification of different water mass and assessing the degree of pollution in a certain aquatic ecosystem. (27)The high concentrations of dissolved oxygen in water bodies could lead to enhanced microbial inactivation by solar UV radiation. (15)Organic matter is a good parameter for measuring the degree of pollution by sewage effluents, organic industrial and agricultural wastes.
Biodegradable organic matter (BOD) in the study area recorded a range exceeding the MPL (60 mg/l) at all sites.Similarly, COD were at very high levels at all sites (Table 1) indicating very high organic loads prevailing in the water of these drains and that these waters required intensive treatment.This agrees with Abdel-Wahaab and Badawy, (2004) (3) , who mentioned that, the water in the drains is currently of poor quality due to pollution from municipal and agriculture sources.
One of the most important impacts of water pollution is the microbial pollution, especially with pathogenic organisms.This is usually caused by the discharge of human fecal wastes into the aquatic environment. (6,28)The bacteriological examination confirmed the results of the physicochemical characterization of the water at the selected sites being highly polluted and needing an immediate solution.This is clearly shown by the high load of the TVC of bacteria obtained in the water samples from the selected canals ( exceeded the MPL of FC (100 cell/100 ml) and were considered fecally polluted.This requires an immediate action For total coliform in water, the selected sites were divided into 2 categories; the first category, sites which were highly polluted (TC content more than 1800 cell/100 ml) and other sites which were less and moderately polluted with TC (less than 300 cell/ 100 ml).FC in the water samples ranged between a minimum of 5 and a maximum of 350 cell/100 ml (Table 2).These low densities of FC in the water doesn't mean that water is safe, but it could be attributed to the detrimental effect of the sunlight in these canals that can lead to significant loss in their numbers in addition to the possible inhibitory effects of the toxic organics that may be available in the water.Sinton et al., (2002) (32) mentioned that sunlight inactivation of fecal  3).Generally, soil showed higher bacterial contamination compared to water samples.
Generally, all the investigated sites exhibited variation in both soil and water content of fecal coliforms.However, when comparing the ratio of soil/water fecal content, it would be clear that the ratio is below two in locations of low pollution (sites 15, 24, 27, 28, 29 and 30), while it is high (ranging from 5 to 360) in locations with high pollution levels (the rest 24 sites).This is attributed to the adsorption of bacterial cells on the soil particles and the availability of organic matter in the soil which provide rich environment for these bacteria. (33)These results agreed with a previous study, (34) where sediments have been shown to harbor fecal coliforms at concentrations higher than those observed in the overlying water column.It was stated that sediments may contain 100 to 1,000 times the number of fecal indicator bacteria contained in the overlying water.
Also Crabill et al., (1999) (35) reported that sediment samples could have up to 2-200 times the FC counts of the water column.
There is an evidence that fecal indicators and pathogenic bacteria don't survive in sediments longer than in the overlying water and it has been proposed that sediments serve as sinks for fecal bacteria with the potential to pollute the overlying bathing waters. (33)e similarity between sampling sites were highly considered as unique (4) Most coliform bacteria are present in large numbers among the normal intestinal flora of humans and other warm-blooded animals, and are thus found in fecal wastes.As a consequence, coliform bacteria, detected in higher concentrations than other pathogenic bacteria, are used as indicators of potential fecal pollution in water environments. (4,7)It is established that these indicators are associated with disease-causing genera of concern to public health. (8)They are also used to measure the sanitary quality of water for recreation, industry, worldwide to measure health hazards. (10)They must be identified in water resources in order to adequately address water problems and protect public health.Therefore, density of such indicators is considered as critical parameter that drives management decision. (11,12)Fecal indicator bacteria in rivers (freshwater supply) were investigated with special attention to the impact of the treatment of waste water effluents on microbiological quality of the receiving waters.A strong negative impact was observed in different areas of the watershed .(13)

Figure 1 :
Figure 1: Sampling sites at Abbis area aliquot sterile distilled water.Bacteria attached to soil particles were thus released and suspended in the distilled water, then serial dilutions were made and used for inoculation.Total viable bacterial counts (TVC) for water and soil samples were determined using pour plate method on nutrient agar medium (Oxoid LTD, Hampshire-England), three replicates were incubated at 37°C for 24 h. (21)The final counts were calculated as colony forming units (CFU/g soil and CFU/ml water) The most probable number (MPN) of total and fecal Coliform bacteria was estimated using MacConkey broth medium (Oxoid LTD, Hampshire-England).Five tubes were used for each of the three concentrations (10.0, 1.0, 0.1 ml) of the sample.The inoculated tubes were incubated at 37 °C for 48 h for the determination of total Coliform bacteria (TC).Another set of tubes were incubated at 44.5°C for 24 h for the determination of fecal Coliform bacteria (FC).Positive reactions were indicated by the production of acid and gas.Positive tubes were streaked on Eosin Methylene blue (EMB) tested for their significance.The similarity between different sampling sites was performed depending on physical, chemical and bacteriological parameters of water samples.The percentages of similarities were plotted into cluster analysis to identify the percentages of closeness of the different sampling sites.The analysis was performed using Minitab version14.

Figure 2 :Figure 3 :
Figure 2: Horizontal distribution of total coliform bacteria (TC) in water at the selected sites in the study area.

Figure 4 :
Figure 4: Horizontal distribution of total coliform bacteria (TC) of the soil in the study area.

Figure 5 :
Figure 5: Horizontal distribution of fecal coliform bacteria (FC) of the soil in the study area.

Figure 6 :
Figure 6: Dendogram, similarity among the sampling sites using similarity coefficient test.
coliforms in fresh water (river) appears to be the most important mechanism inactivating sewage microorganisms in shallow water, although it has been reported to exert different rates of inactivation on various fecal bacteria and bacteriophages.The surviving cells exhibit greater sunlight resistance in natural waters than those from raw sewage Bacteriological assessments of soil in the study area indicated the flourishing of the bacterial populations with high densities of bacterial TVC and heavy contamination with total coliforms derived from domestic sewage that find their way into these canals and drains.This was Bull High Inst Public Health Vol.39 No.3 [2009] confirmed by the results of fecal coliforms where, 15 sites out of the 30 showed very high density of FC indicating recent fecal pollution, and remarkable pollution of the soil in the study area (Table environment from the microbiological and physicochemical point of view.In conclusion, Results of the present study clearly indicated remarkable levels of pollution, these high pollution levels are mainly attributed to the unsanitary disposal of domestic sewage and its mixing with the irrigation water in the different canals which