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Evaluation of microorganisms of drinking water of Rafha City, Northern Borders, Saudi Arabia.

Water is scarce and valuable resource in Saudi Arabia. Thereby, well or ground water is still and will continue to be one of the main sources of drinking water in Saudi Arabia, especially in rural and border areas.

The microbial contamination of drinking water constitutes a major issue worldwide, because it is still a major source of infection and mortality. Evaluation of the microbiological quality of drinking water aims to protect consumers from illness, that due to protozoon's parasites, bacteria, fungi, algae and viruses. Nowadays, prevention of contaminated drinking-water-related-illness is still an important challenge [1].

Rafha-citizens basically obtained their drinking water from treated wells stored in public tanks and re-distributed through waterborne-pipes to houses. However, citizens are usually avoid using tap water as a source of drinking water, owing to their dis-satisfaction of taste, odour and colour of this water. Instead, they prefer drinking waterconsumption of desalinated groundwater obtained from some desalinating-private companies others drink industrially bottled water.

The famous protozoan parasites contaminating drinking water are Giardia, Cryptosporidium, Entamoeba histolytica and Cyclospora, that cause severe gastrointestinal disorders; namely giardiasis, cryptosporidiosis, amoebiasis and cyclosporiasis respectively [2]. Indication of the presence of disease-causing bacteria in drinking water is the coliform bacteria, that are non-pathogenic but associated with many diseases; from which Proteus, Escherichia coli and Enterobacter; indicating faecal contamination of water. Proteus mirabilis is associated with urinary tract infections (3).

In this concept, the coliforms constitute common intestinal commensal bacteria. This group contains important pathogens, as E. coli, which is the most prominent and causes disease when the immune-system is suppressed (4). E. coli considered as an indicator of fecal contamination when found in drinking water (5). Enterobacter species act as opportunistic pathogens.

Many other bacteria are pathogenic, such as Streptococcus that colonizes the heart valves (6), Aeromonas that produces cytotoxic-enterotoxin causes tissue damage (7), Acinetobacter that causes urinary tract infections, pneumonia, endocarditis, wound infections, septicemia and meningitis (8) as well as Pseudomonas that causes bronchopneumonia, ecthyma gangrenosum, urinary tract catheterization, necrotising enterocolitis, hemorrhage, necrosis of the skin (9).

On the other hand, some fungi are pathogenic; such as Aspergillus that causes chronic pulmonary aspergillosis (10), Chrysoporium that cause hyalo-hyphomycosis (11), Alternaria that cause respiratory infections (12) and Acremonium that cause mycetoma, onychomycosis, and hyalohyphomycosis (13).

Some algal genera are pathogenic, as; Anabaena, Microcystis and Oscillatoria. These genera secrete toxins, as Anabaena and Microcystis that secrete microcystin-LR and cylindrospermopsin, leading to liver inflammation, pneumonia, dermatitis, kidney damage and tumor growth and anatoxin-a group (3) -toxin from Anabaena and Oscillatoria that causes nervous disorders (14).

SASO (Saudi Arabian Standards Organization) continuously evaluates drinking water standards for bottled, tap and well-waters to define a quality of water that re-inforce healthy population. These standards set limits for the permissible and maximum contaminants level of parasites and the indicator-microorganism that endanger the health of consumers [15]. A substantial number of these standards are originated from the accurate World Health Organization internationalstandards for drinking water [16].

The main objective of this study is to assess the parasitological and microbiological status of drinkable water of Rafha City in Saudi Arabia.

MATERIALS AND METHODS

Water sampling

The present studies were carried out from July to October, 2014, in Rafha City, Northern Borders, Saudi Arabia. The water-samples had been collected from three major water sources; ten wells, tap and three private desalinating companies; El-Shefaa, Bardy and El-Razaz. Aliquots of 100 ml from each water-sample were collected in sterilized conical flasks, provided with silica gel to keep dryness.

Enumeration of protozoans, bacteria, fungi and algae

Protozoan parasites

All samples preserved in 4% neutralized formalin solution, left to settle [17]. Then, supernatants were collected, filtered through 20 pm net mesh. Materials that retained by filter were then fixed with Lugol's solution. The protozoans were then counted by Sedwgwish Rafter counting method and identified (Cells/100ml), using Olympus binocular compound microscope.

To confirm identification of Giardia lamblia and Cryptosporidium parvum, aliquots of 100 ml of water samples were filtered through a 47 mm diameter, 0.450 im pore size membrane filter. Each material retained by the filter was fixed with Lugol iodine on a separate slide. Fresh preparations of 0.9% saline smear of samples were visually examined for parasitic cysts over approximately 100 fields and then subjected to cold acid-fast. Trichrome staining technique was then applied [18].

Bacteria-count

Coliform bacteria

Coliform bacteria were determined by incubation of samples into lactose broth as presumptive test. The test tubes are placed at 35[degrees]C for 24 hours for gas production. To confirm the presence of coliform, gas produced in incubation into Brilliant Green Bile broth at 35[degrees]C for 24 hours [19]. Water quality analysis was based on the most probable number of Cells/100 ml. The test had been repeated three times.

Aerobic and other facultative anaerobic bacteria

One ml of each water sample was inoculated and spread on blood agar and nutrient agar plates. Then, plates were incubated at 37[degrees]C for 18 to 24 h for determination of bacterial cell count, as cells/100 mL, and for isolation and differentiation of various bacterial strains depending on their morphological descriptions. Different isolated bacteria were further identified by Bilog system (Biolog, Hayward, CA, USA). Experiment had been repeated three times (20).

Fungi

Sabouraud Dextrose Agar (SDA) was used as a culture medium to reveal the presence of fungi and chloramphenicol is used to increase selectivity against commensal microorganisms. For isolation of fungi from water-contaminated specimens, SDA-medium should be inoculated and the plates incubated at 28[degrees]C in an inverted position with increased humidity. For isolation of fungi, two sets of media should be inoculated at 28[degrees]C and a duplicate set at 35[degrees]C. All cultures should be examined 7-10 days for fungal growth, and should be held for 5 weeks before being reported as negative (21). The test had been repeated three times. Algae

Aliquots of 100 ml from each water sample were allowed to settle overnight in sterilized conical flasks. Then after, solid materials from the bottom of the flasks were pipetted for examination. The experiment had been repeated thrice. Algal species had been identified using binocular microscope (Zeiss) (22).

Statistical Methods Analysis of Variance "ANOVA"

The mean data of analysis of variance "ANOVA" between four categories; protozoans, bacteria, fungi and algae, had been achieved to obtain significant differences, using the statistical SPSS-program

RESULTS

Protozoan parasites

The present result showed four protozoan-species, as cysts, Giardia lamblia (Diplomonadida, Hexamitidae), Cryptosporidium parvum (Eucoccidiorida, Cryptosporidiidae), Cyclospora cayatenensis (Eucoccidiorida, Eimeriidae) and Entamoeba coli (Archamoebae).

Table (1) display average counts the highest values in water of the different wells (5.8, 21, 0.7& 15.1 cells/100 ml for G lamblia, C.parvum, C. ayetanensis and E. coli respectively) whereas C. cayetanensis revealed the highest value in tap water, whereas all species were completely absent in water of the three private companies. On the other hand, frequency % revealed the highest values; of G lamblia and E. coli in all sources of water (28.57, 27.27 and 33.33% in well-, tap and companies-water respectively), highest values of C. cayetanensis in tap and the three companies-water (27.27 and 33.33% respectively) and the highest value of C. cayetanensis in well-water (28.57%) (table 2). The statistical analysis of variance "ANOVA" revealed highly significant differences in the two species G lamblia and C. parvum (Table 7(.

Bacteria

Coliform Bacteria

The present work reported four gram-negative coliform bacteria-species; facultative anaerobic Proteus mirabilis, Escherichia coli, Enterobacter cloacae and E. aerogenes.

Table (1) displays mean counted numbers, as CFU/100 ml, where the highest values of P. mirabilis, E. cloacae and E. aerogenes had been found in well-water (1318.7, 34.2 & 92.5 respectively), beside the highest value of E. coli in El-Razaz Company-water (700).

On the other hand, frequency% revealed the highest values; P. mirabilis in well-water (22.73%), P mirabilis and E. aerogenes in tap water (11.11%), P mirabilis in El-Shefaa company (42.86%), E. cloacae in Bardy company (33.33%) and E. coli in El-Razaz company (42.86%) (table 3).

Aerobic Bacteria

Gram negative bacteria

Beside the previous coliform bacteria; aggregated gram negative Acinetobacter haemolyticus and coccobacillus gram-negative Pseudomonas aeruginosa resulted.

The mean counts, as CFU/100 ml, displays the highest values of A. haemolyticus and P aeruginosa in well-water (16.7 & 100000 respectively) (Table 1).

The frequency% revealed the highest values; P aeruginosa in wells (11.36%) and A. haemolyticus in Bardy company (11.11%) (table 4).

Gram positive bacteria

Obligate aerobe Bacillus subtilis, rod shaped Brevibacillus brevis and clusters of spherical

Staphylococcus hominis resulted.

The highest values of mean counts of B. subtilis and B. brevis, as CFU/100 ml, detected in wells (8.3&92.5 respectively) S. hominis in tap water (333.3) (Table 1).

The frequency % scored only B. subtilis and B. brevis in well-water (2.27 & 6.82 respectively) while S. hominis isolated from tap water (11.11) (Table 4).

Anaerobic bacteria

The present results revealed gram positive-facultative species included Bacillus cereus, B. anthracis and Staphylococcus aureus and gram negative-facultative Aeromonas hydrophila and gram-positive-facultative aerobic Streptococcus sanguis.

The highest values of mean counts, as CFU/100 ml, detected B. cereus, S. sanguis and A. hydrophila in wells (11.7, 166.7 & 17.5 respectively), B. anthracis in El-Razaz company-water (66.7) and S. aureus in tap water (333.3) (Table 1).

The detected highest values of frequency %; B. cereus and B. anthracis in El-Razaz company-water (14.29 & 42.86 respectively) whereas S. aureus, S. sanguis and A. hydrophila isolated from tap water (33.33), El-Shefaa company-water (42.86) and well-water (6.82) (Table 4).

Regarding bacterial counts, as cells/100 ml, the statistical analysis of variance "ANOVA" revealed highly significant differences in the coliform bacteria Proteus mirabilis and Enterobacter cloacae (Table 7).

Fungi

The present mycoflora that isolated from drinking water of the target three sources are nine genera; Aspergillus fumigatus, Aspergillus flavus, Chrysoporium tropicum, Alternaria alternate, Acremonium sp., Penicillium sp., Cladosporium cladosporioides, Rhodotorula mucilaginosa and Exophiala jeanselmei-yeast.

Table (1) revealed that 8, 4, 2, 3 and 3 colonies of fungal genera were isolated from the present three water-sources; wells, tap, El-Shefaa, Bardy & El-Razaz company respectively. The highest values of mean counts of fungal genera, as CFU/100 ml, were for Aspergillus flavus (3.3, 4 & 2.3 for wells, El-Shefaa and Bardy companies respectively), Alternaria alternate (20.3 for tap water) and Penicillium (4 for El-Razaz).

It is worthy of mentioning that the fungus A. flavus appeared in all five water-sources followed by A. alternate in four sources.

Concerning counts of the fungi, as CFU/ 100ml, the statistical analysis of variance "ANOVA" revealed low significant differences between the four sources; wells, tap and the three companies concerning Aspergillus fumigatus and Penicillium whereas rest of fungi show non-significant differences (Table 7).

Algae

Six algal genera had been isolated from four different water-sources except wells; Merismopedia tenuissima, Anabaena sphaerica, Microcystis aeruginosa, Oscillatoria limnetica and diatoms, including; Cyclotella sp., Synedra sp., Melosira sp., Fragilaria sp., Cymbella sp. and Nitzschia sp.

The highest values of the resulted mean counts of algae, as cells/100 ml, included; M. tenuissima from El-Shefaa and Bardy companies-water (10.7 for both), A. sphaerica, and two diatoms Melosira and Fragilaria from tap water (0.3, 5 & 6 respectively). The collected species from El-Razaz company-water included; O. limnetica (0.7), the diatoms Cymbella sp. and Fragilaria sp. from tap water (4.3 & 6 respectively). M. aeruginosa and the diatoms Cyclotella sp., Synedra sp., Melosira sp. and Nitzschia sp. from tap water (7, 3, 5, 5.3 & 0.3 respectively) (Table 1).

The highest values of frequency % of algae-samples are; in tap and El-Shefaa and Bardy companies-water forM. tenuissima (1.7, 10, 10.7 and 10.7) and in El-Razaz company-water for Oscillatoria limnetica (0.7) (Table 6).

Respecting algal counts "cells/100 ml", the statistical analysis of variance revealed highly significant differences in the alga M. tenuissima between the present four sources (Table 7).

DISCUSSION

The present work revealed several pathogenic microorganisms in three sources of water; 10 wells, tap and three private desalinating water-companies; including three parasitic protozoan-cysts, three genera of Coliform bacteria-group and some aerobic and anaerobic bacteria, five genera of fungi as well as three genera of algae.

In this respect, results of Al-Turki (23) reinforced the present results where he revealed that microbiological water quality results showed that 20% of the samples examined are contaminated with coliform bacteria (Escherichia coli, and Enterobacter aerogenes), indicating the necessity of water-sanitation of Hael's water prior to use.

WHO estimated that 1.8 million people die each year as a result of severe diarrhea caused by drinking contaminated water, where developing countries have the prevalence of giardiasis in patients with diarrhea is about 20% (24).

Concerning parasitic cysts of Giardia lamblia, Cryptosporidium parvum and Cyclospora cayatenensis, they contaminate water-sources causing severe gastrointestinal disorders. The maximum score of the present parasitic protozoans is for C. parvum and minimum count for C. cayetanensis.

Giardia-cysts resist chlorine leads to giardiasis, its symptoms are flu and severe gastrointestinal disorders, detected in 81% of raw water samples and 17% of filtered water samples in the United States (25). In this respect, WHO declared that chlorine disinfection of drinking-water has limitations against the protozoan pathogens, in particular Cryptosporidium (26). C. parvum is protozoan parasite causes Cryptosporidiosis in humans. Oocysts of C. parvum in 87% of raw water samples and 27% of drinking water samples in 15 Canadian regions (25). Le Chevallier et al. reinforced the present counts of C. parvum-cysts which is higher than those of G. lamblia in all water sources (25).

Cyclospora cayetanensis is cyst-forming coccidian protozoan that causes a self-limiting diarrhea named cyclosporiasis, with symptoms range from watery, loose stool, weight loss, cramping, fatigue, vomiting, fever and nausea [2].

The maximum score of coliform bacteria are for Proteus and the minimum for Enterobacter. Proteus species causes wound infections, septicemia and pneumonia whereas E. coli produce potentially lethal toxins and causes food poisoning. Moreover, Uro-pathogenic E. coli is responsible for 90% of urinary tract infections [27]. Enterobacter causes opportunistic infections, where the urinary and respiratory tracts are the sites of infection (28).

Concerning other gram positive bacteria, Acinetobacter haemolyticus causes pneumonia, bacteremia and meningitis (29), Pseudomonas aeruginosa infects urinary tract, burns, wounds and causes blood infections (9).

Gram positive bacteria, included Bacillus subtilis and Staphylococcus hominis cause diseases in severely immune-compromised patients, Bacillus cereus causes severe nausea, vomiting, and diarrhea, Bacillus anthracis leads to anthrax disease (30) Staphylococcus aureus causes scalded-skin syndrome (31), Streptococcus sanguis causes sub-acute bacterial endocarditis (32) and Aeromonas hydrophila produces aerolysin cytotoxic enterotoxin leads to tissue damage (9).

The studies revealed 5 pathogenic fungal species isolated. The maximum count of fungal-species are for Aspergillus flavus and minimum for Cladosporium cladosporioides.

The most common pathogenic species are Aspergillus fumigatus and A. flavus. The latter species produces aflatoxin-toxin that contaminating foods and considers carcinogen leads to allergic disease while A. fumigatus causes allergic disease leads to chronic pulmonary infections (33). Alternaria alternate causes respiratory infections and asthma in humans with compromised immunity (34).

Hyalo-hyphomycosis that caused by Acremonium includes arthritis, osteomyelitis, peritonitis, endocarditis, pneumonia, cerebritis, and subcutaneous infection (35).

Exophiala jeanselmei causes mycetoma, localized cutaneous infections, subcutaneous cysts, endocarditis, cerebral and disseminated infections, beside phaeohyphomycosis (36).

The highest value of the algae are for Merismopedia tenuissima and minimum for Anabaena sphaerica. Anabaena sphaerica and Microcystis aeruginosa secrete both microcystinLR and cylindrospermopsin toxins that leading to liver inflammation, pneumonia, dermatitis, kidney damage and tumor growth. Anatoxin-a group3-toxin is also secreted by Anabaena and Oscillatoria limnetica, that causes nervous disorders (14).

Concerning standard levels of microorganisms in drinking water, EPA's MCLG declared that detection of protozoans, especially Cryptosporidium, is difficult and not technically feasible for routine analysis of human drinking water. The recommendation considered the proper way to control pathogenic protozoans is using an effective water treatment technique, such as reverse osmosis or ozonation.

According to EPA MCL, coliform must be less than one/100 mL. In this concern, the microbiological guidelines and standards for drinking water for E. coli are zero CPU count/100 ml (37). In this respect, average count/100 mL of all water sources of the present work revealed considerable higher levels than those of EPA MCL and an apparent risk on Rafha's citizens-health.

The present investigation indicated that water sources of Rafha city are important contributor to transmission of contaminants to consumers.

The present work recommends a future-field-application of selected specialized anti-parasites and anti-microbial to control the present resulting contaminators to gain considerable safe levels of drinking water.

ACKNOWLEDGMENTS

The authors would like to acknowledge the approval and the support of this research study by the Deanship of the Scientific Research, Northern Border University, Arar, KSA and thanks to Dean of Faculty of Science and Arts, Rafha. The authors want to extend our appreciation to Doaa Abd Allah and Doaa Ibrahim for review of this research.

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A.A. Abdel Haleem [1,2], S.K. Hemida [1,3] and M.M. Abdellatif [1,4]

[1] Department of Biology, Faculty of Arts & Science, Northern Border University, Rafha, Saudi Arabia,

[2] Department of Biological sciences, Faculty of Education, Ain Shams University, Cairo, Egypt.

[3] Department of Botany, Faculty of Science, Assiut University, Egypt.

[4] Department of Microbiology, College of Veterinary Medicine, Nyala University, Sudan.

(Received: 10 November 2015; accepted: 11 January 2016)

* To whom all correspondence should be addressed. Tel.: +966508379140;

E-mail: abdelsalam1958@windowslive.com
Table 1. Average counted-number of protozoan parasites,
bacteria, fungi and algae (mean count/100 ml) which obtained
from water-resources of Rafha city, Saudi Arabia

                                     Average count/100 ml

Species   Total isolated-species      Wells    Tap water
No.

          I. Protozoan parasites:
1         Giardia lamblia             5.8          3
2         Cryptosporidium parvum      21          17
3         Cyclospora cayetanensis     0.7         0.7
4         Entameba coli               15.1        7.3
          II. Coliform
          bacteria-group
          "F acultative Anaerobes":
          Proteus mirabilis
1         Escherichia coli            1318.7     33.3
2         Enterobacter cloacae        192.5       370
3         E. aerogenes                34.2         0
4         III. Total bacteria:        92.5        3.3
          a) Aerobic bacteria:
          Bacillus subtilis
1         Brevibacillus brevis        8.3          0
2         Staphylococcus hominis      92.5         0
3         Acinetobacter               0          333.3
          haemolyticus
4         b) Other facultative        16.7        3.3
          anaerobic Bacteria:
          Bacillus cereus
          Bacillus anthracis
5         Staphylococcus aureus       11.7         0
6         Streptococcus sanguis       0.8        33.3
7         Aeromonas hydrophila        166.7      333.3
8         Pseudomonas aeruginosa      166.7        0
9         IV. Fungi:                  17.5         0
10        Aspergillus fumigatus       100000    0.33333
          Aspergillus flavus
1         Chrysoporium tropicum       1.3          0
2         Alternaria alternate        3.3          3
3         Acremonium sp.              1.1          0
4         Penicillium sp.             0.9        20.3
5         Cladosporium                1           1.3
          cladosporioides
6         Rhodotorula mucilaginosa    0            0
7         Exophiala jeanselmei        0.1          0
8         V. Algae:                   0.3          0
9         Diatoms                     0.7         0.3
          Cyclotella sp.
1         Synedra sp.
1         Melosira sp.                0            3
2         Cymbella sp.                0            5
3         Fragilaria sp.              0           5.3
4         Nitzschia sp.               0           4.3
5         Merismopedia tenuissima     0            6
6         Anabaena sphaerica          0           0.3
2         Microcystis aeruginosa      0           10
3         Oscillatoria limnetica      0           0.3
4                                     0           0.7
5                                     0            0

                                         Average count/100 ml

                                           Private companies

Species   Total isolated-species      El-Shefaa   Bardy   El-Razaz
No.

          I. Protozoan parasites:
1         Giardia lamblia                1.3        2       2.3
2         Cryptosporidium parvum         4.3       7.7      7.3
3         Cyclospora cayetanensis         0         0        0
4         Entameba coli                   6        12       10.7
          II. Coliform
          bacteria-group
          "F acultative Anaerobes":
          Proteus mirabilis
1         Escherichia coli               35         0        0
2         Enterobacter cloacae          33.3      66.7      700
3         E. aerogenes                    0        6.7       0
4         III. Total bacteria:            0       36.7       0
          a) Aerobic bacteria:
          Bacillus subtilis
1         Brevibacillus brevis            0         0        0
2         Staphylococcus hominis          0         0        0
3         Acinetobacter                   0         0        0
          haemolyticus
4         b) Other facultative           3.3        0        0
          anaerobic Bacteria:
          Bacillus cereus
          Bacillus anthracis
5         Staphylococcus aureus          3.3       3.3      3.3
6         Streptococcus sanguis           0         0       66.7
7         Aeromonas hydrophila            0         0        0
8         Pseudomonas aeruginosa        33.3        0        0
9         IV. Fungi:                      0         0        0
10        Aspergillus fumigatus        0.33333      0        0
          Aspergillus flavus
1         Chrysoporium tropicum           0        0.7      0.3
2         Alternaria alternate            4        2.3      3.3
3         Acremonium sp.                  0         0        0
4         Penicillium sp.                1.3       1.7       0
5         Cladosporium                    0         0        0
          cladosporioides
6         Rhodotorula mucilaginosa        0         0        4
7         Exophiala jeanselmei            0         0        0
8         V. Algae:                       0         0        0
9         Diatoms                         0         0        0
          Cyclotella sp.
1         Synedra sp.
1         Melosira sp.                    0         0        0
2         Cymbella sp.                    0         0        0
3         Fragilaria sp.                  0        0.3       0
4         Nitzschia sp.                   0         0       0.7
5         Merismopedia tenuissima        1.3       0.3       0
6         Anabaena sphaerica             0.3        0        0
2         Microcystis aeruginosa        10.7        0       0.7
3         Oscillatoria limnetica          0         0        0
4                                         0         0        0
5                                         0        0.7      0.3

Table 2. Frequency% of protozoan parasites in different water
sources, Rafha city, Northern Borders, Saudi Arabia

Types of        Giardia   Cryptosporidium    Cyclospora    Entamoeba
water           lamblia       parvum        cayatenensis     coli

Well water       28.57         14.29           28.57         28.57
Tap water        27.27         27.27           18.18         27.27
El-Shefaa Co.    33.33         33.33           00.00         33.33
Bardy Co.        33.33         33.33           00.00         33.33
El-Razaz Co.     33.33         33.33           00.00         33.33

Table 3. Frequency% of coliform-group bacteria in different
water sources, Rafha city, Northern Borders, Saudi Arabia

                      Isolated coliform groups "negative gram"

Types of         Proteus    Escherichia   Enterobacter   Enterobacter
water           mirabilis      coli         cloacae       aerogenes

Well water        22.73        13.64         09.09          06.82
Tap water         11.11        33.33         00.00          11.11
El-Shefaa Co.     42.86        14.29         00.00          00.00
Bardy Co.         00.00        22.22         33.33          22.22
El-Razaz Co.      00.00        42.86         00.00          00.00

Table 4. Frequency % of aerobic and anaerobic bacteria in
different water sources, Rafha city, Northern Borders, Saudi Arabia

                        Anaerobic bacteria

Types of        Bacillus     Bacillus      Staphylococcus
water            cereus      anthracis         aureus

Well water       13.64         2.27             00.00
Tap water        00.00         00.00            33.33
El-Shefaa Co.    00.00         00.00            00.00
Bardy Co.        11.11         00.00            00.00
El-Razaz Co.     14.29         42.86            00.00

                            Anaerobic bacteria

Types of        Streptococcus   Aeromonas    Pseudomonas
water              sanguis      hydrophila   aeruginosa

Well water          00.00          6.82         11.36
Tap water           00.00         00.00         00.00
El-Shefaa Co.       42.86         00.00         00.00
Bardy Co.           00.00         00.00         00.00
El-Razaz Co.        00.00         00.00         00.00

                       Aerobic bacteria

Types of           Bacillus      Brevibacillus
water              subtilis         brevis

Well water           2.27            6.82
Tap water           00.00            00.00
El-Shefaa Co.       00.00            00.00
Bardy Co.           00.00            00.00
El-Razaz Co.        00.00            00.00

                       Aerobic bacteria

Types of        Staphylococcus   Acinetobacter
water              hominis       haemolyticus

Well water          00.00            4.55
Tap water           11.11            00.00
El-Shefaa Co.       00.00            00.00
Bardy Co.           00.00            11.11
El-Razaz Co.        00.00            00.00

Table 5. Frequency % of fungi in different water sources,
Raflia city. Northern Borders, Saudi Arabia.

Types of         Aspergillus   Aspergillus   Chrysoporium
water             fumigatus      flavus        tropicum

Well water          38.89         22.22         16.67
Tap water           00.00         12.50         00.00
El-Shefaa Co.       00.00         50.00         00.00
Bardy Co.           25.00         50.00         00.00
El-Razaz Co.        20.00         40.00         00.00

Types of         Alternaria   Acremonium   Petiicillium
water            alternate

Well water         05.56        05.56         00.00
Tap water          37.50        25.00         00.00
El-Shefaa Co.      25.00        00.00         00.00
Bardy Co.          25.00        00.00         00.00
El-Razaz Co.       00.00        00.00         40.00

Types of          Cladosporium     Rhodotorula    Phialophora
water            cladosporioides   mucilaginosa

Well water            05.56           03.00          00.00
Tap water             00.00           00.00          12.50
El-Shefaa Co.         00.00           00.00          00.00
Bardy Co.             00.00           00.00          00.00
El-Razaz Co.          00.00           00.00          00.00

Table 6. Frequency % of algae in different water sources,
Rafha city, Northern Borders, Saudi Arabia

Types of        Merismopedia   Anabaena    Microcystis   Oscillatoria
water            tenuissima    sphaerica   aeruginosa     limnetica

Well water          00.0         00.0         00.0           00.0
Tap water           10.0         00.3         00.7           00.0
El-Shefaa Co.       10.7         00.0         00.0           00.0
Bardy Co.           10.7         00.0         00.0           00.0
El-Razaz Co.        00.0         00.0         00.0           00.7

                              Diatoms
Types of
water           Cyclotella    Synedra     Melosira

Well water         00.0         00.0        00.0
Tap water          03.0         05.0        05.0
El-Shefaa Co.      00.0         00.0        00.0
Bardy Co.          00.0         00.0        00.0
El-Razaz Co.       00.0         00.0        00.3

                              Diatoms
Types of
water            Cymbella    Fragilaria   Nitzschia

Well water         00.0         00.0        00.0
Tap water          04.0         06.0        00.3
El-Shefaa Co.      00.0         01.3        00.3
Bardy Co.          00.0         01.3        00.3
El-Razaz Co.       00.0         00.3        00.0

Table 7. Statistical analysis of variance (ANOVA)
of counts of protozoan parasites, bacteria, fungi
and algae-species (CFU or cells/100 ml water), from
different water sources of Rafha City, Saudi Arabia
(N.S = non-significant - P>0.001 = highly significant)

No    Source of variation    Species                   df   SS

I-    Protozoan parasites:
1       Between 5 sources    Giardia lamblia           4    35.19
2       "Wells,              Cryptosporidium parvum    4    611.88
3       Tap,                 Cyclospora cayetanensis   4    1.62
4       El-Shefaa,           Entamoeba coli            4    159.45
        Bardy & El-Razaz     II- Bacteria:
        Private Companies'

a)    Coliform bacteria "facultative anaerobes":
1                            Proteus mirabilis         4    4773169
2                            Escherichia coli          4    232239
3                            Enterobacter cloacae      4    2411.34
4                            Enterobacter aerogenes    4    18464.6

b)    Aerobic bacteria:
5                            Bacillus subtilis         4    166.53
6                            Brevibacillus brevis      4    20535
7                            Staphylococcus hominis    4    266661
8                            Acinetobacter             4    573.63
                             haemolyticus

c)    Other facultative anaerobic bacteria:
1                            Bacillus cereus           4    2026.69
2                            Bacillus anthracis        4    10569.4
3                            Staphylococcus aureus     4    266663
4                            Streptococcus sanguis     4    62669.3
5                            Aeromonas hydrophila      4    735
6                            Pseudomonas aeruginosa    4    2.36 X
                                                            [10.sup.11]
                             III- Fungi:
1                            Aspergillus fumigatus     4    3.32
2                            Aspergillus flavus        4    4.46
3                            Chrysoporium tropicum     4    2.8
4                            Alternaria alternate      4    903.3
5                            Acremonium                4    5.05
6                            Penicillium               4    38.4
7                            Cladosporium              4    0.26
                             cladosporioides
8                            Total yeasts              4    8062.66
9                            Exophiala jeanselmei      4    1.07

1                            Total Diatoms             4    287.17
2                            Merismopedia tenuissima   4    333.59
3                            Anabaena sphaerica        4    0.25
4                            Microcystis aeruginosa    4    0.94
5                            Oscillatoria limnetica    4    0.95

No    Source of variation    Species                   MS

I-    Protozoan parasites:
1       Between 5 sources    Giardia lamblia           17.6
2       "Wells,              Cryptosporidium parvum    305.94
3       Tap,                 Cyclospora cayetanensis   0.81
4       El-Shefaa,           Entamoeba coli            79.73
        Bardy & El-Razaz     II- Bacteria:
        Private Companies'

a)    Coliform bacteria "facultative anaerobes":
1                            Proteus mirabilis         2386585
2                            Escherichia coli          116120
3                            Enterobacter cloacae      1205.67
4                            Enterobacter aerogenes    9232.32

b)    Aerobic bacteria:
5                            Bacillus subtilis         83.27
6                            Brevibacillus brevis      10267.5
7                            Staphylococcus hominis    133331
8                            Acinetobacter             286.81
                             haemolyticus

c)    Other facultative anaerobic bacteria:
1                            Bacillus cereus           1013.35
2                            Bacillus anthracis        5284.69
3                            Staphylococcus aureus     133332
4                            Streptococcus sanguis     31334.7
5                            Aeromonas hydrophila      367.5
6                            Pseudomonas aeruginosa    1.18 X
                                                       [10.sup.11]
                             III- Fungi:
1                            Aspergillus fumigatus     1.66
2                            Aspergillus flavus        2.23
3                            Chrysoporium tropicum     1.4
4                            Alternaria alternate      451.65
5                            Acremonium                2.53
6                            Penicillium               19.2
7                            Cladosporium              0.13
                             cladosporioides
8                            Total yeasts              4031.33
9                            Exophiala jeanselmei      0.54

1                            Total Diatoms             143.58
2                            Merismopedia tenuissima   166.79
3                            Anabaena sphaerica        0.12
4                            Microcystis aeruginosa    0.47
5                            Oscillatoria limnetica    0.48

No    Source of variation    Species                   F

I-    Protozoan parasites:
1       Between 5 sources    Giardia lamblia           19.49
2       "Wells,              Cryptosporidium parvum    19.92
3       Tap,                 Cyclospora cayetanensis   8.02
4       El-Shefaa,           Entamoeba coli            6.96
        Bardy & El-Razaz     II- Bacteria:
        Private Companies'

a)    Coliform bacteria "facultative anaerobes":
1                            Proteus mirabilis         13.87
2                            Escherichia coli          2.18
3                            Enterobacter cloacae      48.34
4                            Enterobacter aerogenes    5.62

b)    Aerobic bacteria:
5                            Bacillus subtilis         2.4
6                            Brevibacillus brevis      2.47
7                            Staphylococcus hominis    2.4
8                            Acinetobacter             6.26
                             haemolyticus

c)    Other facultative anaerobic bacteria:
1                            Bacillus cereus           1.68
2                            Bacillus anthracis        4.75
3                            Staphylococcus aureus     2.26
4                            Streptococcus sanguis     7.78
5                            Aeromonas hydrophila      9.54
6                            Pseudomonas aeruginosa    1.97

                             III- Fungi:
1                            Aspergillus fumigatus     4.74
2                            Aspergillus flavus        0.18
3                            Chrysoporium tropicum     2.39
4                            Alternaria alternate      2.27
5                            Acremonium                2.63
6                            Penicillium               5.49
7                            Cladosporium              2.35
                             cladosporioides
8                            Total yeasts              1.07
9                            Exophiala jeanselmei      1.93

1                            Total Diatoms             0.35
2                            Merismopedia tenuissima   11.77
3                            Anabaena sphaerica        2.08
4                            Microcystis aeruginosa    8.48
5                            Oscillatoria limnetica    4.29

No    Source of variation    Species                   P-value

I-    Protozoan parasites:
1       Between 5 sources    Giardia lamblia           0.001> P
2       "Wells,              Cryptosporidium parvum    0.001> P
3       Tap,                 Cyclospora cayetanensis   0.01> P
4       El-Shefaa,           Entamoeba coli            0.01> P
        Bardy & El-Razaz     II- Bacteria:
        Private Companies'

a)    Coliform bacteria "facultative anaerobes":
1                            Proteus mirabilis         0.001> P
2                            Escherichia coli          N. S
3                            Enterobacter cloacae      0.001> P
4                            Enterobacter aerogenes    0.05> P

b)    Aerobic bacteria:
5                            Bacillus subtilis         N. S
6                            Brevibacillus brevis      N. S
7                            Staphylococcus hominis    N. S
8                            Acinetobacter             0.01> P
                             haemolyticus

c)    Other facultative anaerobic bacteria:
1                            Bacillus cereus           N. S
2                            Bacillus anthracis        0.05> P
3                            Staphylococcus aureus     N. S
4                            Streptococcus sanguis     0.01> P
5                            Aeromonas hydrophila      0.01> P
6                            Pseudomonas aeruginosa    N. S

                             III- Fungi:
1                            Aspergillus fumigatus     0.05> P
2                            Aspergillus flavus        N. S
3                            Chrysoporium tropicum     N. S
4                            Alternaria alternate      N. S
5                            Acremonium                0.10> P
6                            Penicillium               0.05> P
7                            Cladosporium              N. S
                             cladosporioides
8                            Total yeasts              N. S
9                            Exophiala jeanselmei      N. S
                                                       IV- Algae:
1                            Total Diatoms             N. S
2                            Merismopedia tenuissima   0.001> P
3                            Anabaena sphaerica        N. S
4                            Microcystis aeruginosa    0.01> P
5                            Oscillatoria limnetica    0.05> P
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Article Details
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Author:Haleem, A.A. Abdel; Hemida, S.K.; Abdellatif, M.M.
Publication:Journal of Pure and Applied Microbiology
Article Type:Report
Geographic Code:7SAUD
Date:Mar 1, 2016
Words:6079
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