Significance of Fungal Species Isolated from Blue Nile River and Tuti Island on Drinking Water Quality

Aims: This study has been carried out with the following objectives: Assessment of contamination of water in Blue Nile through fungi of different water samples collected and Identify, as far as possible, the species responsible for contamination and, to a diver these problems. Study respectively. While the isolated genera from Blue Nile water that inoculated in PDA + Chloramphenicol were three as Fusarium spp., Aspergillus niger and Candida albicans with percentage of 70%, 20%, and 10% respectively. Also three genera were isolated from tap water that inoculated in PDA + Chloramphenicol and Rose Bengal as Mucor spp., Aspergillus fumigatus , and Rhizopus spp. and the frequencies are (72.73%, 18.2%, and 9.1%) respectively. The isolated genera from tap water which inoculated in PDA + Chloramphenicol show Mucor spp. (57.14%), Rhizopus spp., Aspergillus fumigatus and Aspergillus terrus with frequency of 14.29% for all three lateral genera. Most genera were shown sensitive when subjected to the different concentrations of chlorine (0.2, 0.5, and 0.7 g/ 50 ml) except Candida albicans , Fusarium species, and Aspergillus niger were shown resist to chlorine concentration 0.5 g/ 50 ml. Conclusion: The recovers of fungi from drinking water indicates low levels of quality and need intensive treatment.


INTRODUCTION
The Nile River is the main drinking water resource for the Sudanese people providing %97 of their water. The problem of water quality is especially acute for developing world where up to %90 of cities discharge their untreated sewages into rivers and streams. The problem of air pollution is continuing to grow more acute in the developing world population are expanding rapidly and rapid industrialization is complied with increasing use of motor vehicles the air condition and fresh water quality in the area around the Nile river have been severely affected. The impact of pollution in Sudan appears in all environments [1]. The contaminants that are found in drinking water are bacteria, viruses, pathogenic protozoa, algae and fungi (yeast and mould) are important causes of water borne diseases [2]. Fungi in water are becoming more important because of increasing number of immune -suppressed patients, nonetheless water borne fungi are associated with taste and odour problem. Study of water borne fungi are low because primary fungi histoplasma, coccidiodes, blastomycete, paracoccidiodes are not known as water borne. However there is a number of opportunistic human fungi pathogen which are known to be water borne and several studies have suggest that inhalation water isolated aerosols containing Spores of these fungi may be route in systematic infection in human [3]. Furthermore some fungal species from water supplies are potentially allergenic or toxigenic [4]. This study has been carried out with the following objectives: Assessment of contamination of water in Blue Nile through fungi of different water samples collected and Identify, as far as possible, the species responsible for contamination and, to a diver these problems.

Collection of Sample from Blue Nile River
The water sample were collected from Blue Nile as follow: 180 ml of water sample were collected in sterile clear glass container put at the angle 45º and opened under water for distance 20 cm, filled and small spaces were lift for shaking and screwed, then the bottles were transported to the laboratory using ice-bag within two hours [5].

Collection of Water Sample from Tap Water
The water sample were collected from taps as follow: 180 ml of water sample were collected in sterile clear glass bottle contained sodium thiosulphate to remove residual chlorine, the tap was cleaned by tissue and alcohol to remove the dust and sterilize by flaming, the water was flowed for one minutes before sampling without touch or contaminate cap, the sample bottle was carefully opened and filled, a small spaces were lift for shaking and screwed, the sample was transported to the laboratory using ice-bag within two hours [5].

Isolation of Fungi from Water Sources
Ten water samples from Blue Nile River and a ten water sample from tap were collected. The samples were inoculated onto sterilized potato dextrose agar in two batches, batch one supplemented with rose Bengal and chloramphenicol while batch two supplemented with chloramphenicol only. By taken 0.1 ml of water sample and inoculated onto the surface of sterilize plates, then the plates were incubated at 28°C for 5 days [6].

Effect of the Chlorine in Fungal Growth
To study the effect of chlorine in fungal growth, the twenty samples were treated with 0.2, 0.5, 0.7 g chlorine respectively. These concentrations were added to 50 ml of water sample and let for 1 hour.

Isolated Fungi Inoculated in PDA + Chloramphenicol + Rose Bengal (Blue Nile River)
The dominant genera isolated from Blue Nile River are the Fusarium species which shown in 5 sample out of twelve (41.47%) followed by Aspergillus fumigatus and Candida albicans which isolated from 3 sample (25%), finally, Aspergillus niger shown in one sample (8.33%) Fig. 1.

Isolated Fungi Inoculated in PDA + Chloramphenicol (Blue Nile River)
Fusarium species were also shown in 7 outgrow of ten sample collected (70%), followed by Aspergillus niger which was isolated from 2 sample (20%) and Candida albicans which observed in one sample (10%) Aspergillus fumigatus did not shown in this medium Fig. 2.

Effect of the Chlorine in Fungal Growth
To study the effect of chlorine in fungal growth, the twenty samples were treated with 0.2, 0.5, 0.7 g chlorine respectively. These concentrations were added to 50 ml of water sample and let for 1 hour.

Area of Study
This study was conducted at the Department of Microbiology and Molecular Biology, Faculty of Science and Technology, Al-Neelain University, Khartoum -Sudan. All the experiments were accomplished aseptically in the Laboratory of Microbiology.

Isolated Fungi Inoculated in PDA + Chloramphenicol + Rose Bengal (Blue Nile River)
The dominant genera isolated from Blue Nile River are the Fusarium species which shown in 5 sample out of twelve (41.47%) followed by Aspergillus fumigatus and Candida albicans which isolated from 3 sample (25%), finally, Aspergillus niger shown in one sample (8.33%) Fig. 1.

Isolated Fungi Inoculated in PDA + Chloramphenicol (Blue Nile River)
Fusarium species were also shown in 7 outgrow of ten sample collected (70%), followed by Aspergillus niger which was isolated from 2 sample (20%) and Candida albicans which observed in one sample (10%) Aspergillus fumigatus did not shown in this medium Fig. 2.

Effect of the Chlorine in Fungal Growth
To study the effect of chlorine in fungal growth, the twenty samples were treated with 0.2, 0.5, 0.7 g chlorine respectively. These concentrations were added to 50 ml of water sample and let for 1 hour.

Isolated Fungi Inoculated in PDA + Chloramphenicol + Rose Bengal (Blue Nile River)
The dominant genera isolated from Blue Nile River are the Fusarium species which shown in 5 sample out of twelve (41.47%) followed by Aspergillus fumigatus and Candida albicans which isolated from 3 sample (25%), finally, Aspergillus niger shown in one sample (8.33%) Fig. 1.

Isolated Fungi Inoculated in PDA + Chloramphenicol (Blue Nile River)
Fusarium species were also shown in 7 outgrow of ten sample collected (70%), followed by Aspergillus niger which was isolated from 2 sample (20%) and Candida albicans which observed in one sample (10%) Aspergillus fumigatus did not shown in this medium Fig. 2.

Isolated Fungi Inoculated in PDA+ Chloramphenicol +Rose Bengal (Tap Water)
The dominant mould isolated from tap water are the Mucor species which was shown in 8 samples (72.73%) followed by Aspergillus fumigatus which was shown in two samples (18.2%) and Rhizopus stolonifier shown in one sample (9.1%) Fig. 3.

Isolated Fungi Inoculated in PDA+ Chloramphenicol Only (Tap Water)
Mucor species also appeared as the predominant mould that isolated from four samples (57.14%), followed by Rhizopus stolonifier, Aspergillus fumigatus, and Aspergillus terrus which were shown on one sample (14.29%) Fig. 4.
The outcome of this study is similar to the Warris et al. [7], who reported that the dominant fungi isolated from water are Fusarium and Aspergillus species, and it is resemble to that of Denning et al. [8], and Straus, [9], whom reported that the water are contaminated by microorganisms such as Aspergillus fumigatus, Mucor, Abisidia, and Candida albicans.
The percentage of these fungi isolated it is semi-like to Rukaia et al. [10] who reported that the percentage of finding fungi as the follow A. niger 27% this is more than the outcome of our study (20%), A. fumigatus 24.1% which is almost similar of our finding, Fusarium spp. 41.47% this is more than the outcome of our study (25%), Rhizopus spp. 14.29% shown less than isolated species in this study (20.4%), and Mucor spp. are not isolated during the investigation of Rukaia et al. [10] who reported that the percentage of Mucor spp. is zero percent (0.00%) which is in contradiction with our finding as dominant fungi that isolated from the tap water (72.73%).

Isolated Fungi Inoculated in PDA+ Chloramphenicol +Rose Bengal (Tap Water)
The dominant mould isolated from tap water are the Mucor species which was shown in 8 samples (72.73%) followed by Aspergillus fumigatus which was shown in two samples (18.2%) and Rhizopus stolonifier shown in one sample (9.1%) Fig. 3.

Isolated Fungi Inoculated in PDA+ Chloramphenicol Only (Tap Water)
Mucor species also appeared as the predominant mould that isolated from four samples (57.14%), followed by Rhizopus stolonifier, Aspergillus fumigatus, and Aspergillus terrus which were shown on one sample (14.29%) Fig. 4.
The outcome of this study is similar to the Warris et al. [7], who reported that the dominant fungi isolated from water are Fusarium and Aspergillus species, and it is resemble to that of Denning et al. [8], and Straus, [9], whom reported that the water are contaminated by microorganisms such as Aspergillus fumigatus, Mucor, Abisidia, and Candida albicans.
The percentage of these fungi isolated it is semi-like to Rukaia et al. [10] who reported that the percentage of finding fungi as the follow A. niger 27% this is more than the outcome of our study (20%), A. fumigatus 24.1% which is almost similar of our finding, Fusarium spp. 41.47% this is more than the outcome of our study (25%), Rhizopus spp. 14.29% shown less than isolated species in this study (20.4%), and Mucor spp. are not isolated during the investigation of Rukaia et al. [10] who reported that the percentage of Mucor spp. is zero percent (0.00%) which is in contradiction with our finding as dominant fungi that isolated from the tap water (72.73%).

Isolated Fungi Inoculated in PDA+ Chloramphenicol +Rose Bengal (Tap Water)
The dominant mould isolated from tap water are the Mucor species which was shown in 8 samples (72.73%) followed by Aspergillus fumigatus which was shown in two samples (18.2%) and Rhizopus stolonifier shown in one sample (9.1%) Fig. 3.

Isolated Fungi Inoculated in PDA+ Chloramphenicol Only (Tap Water)
Mucor species also appeared as the predominant mould that isolated from four samples (57.14%), followed by Rhizopus stolonifier, Aspergillus fumigatus, and Aspergillus terrus which were shown on one sample (14.29%) Fig. 4.
The outcome of this study is similar to the Warris et al. [7], who reported that the dominant fungi isolated from water are Fusarium and Aspergillus species, and it is resemble to that of Denning et al. [8], and Straus, [9], whom reported that the water are contaminated by microorganisms such as Aspergillus fumigatus, Mucor, Abisidia, and Candida albicans.
The percentage of these fungi isolated it is semi-like to Rukaia et al. [10] who reported that the percentage of finding fungi as the follow A. niger 27% this is more than the outcome of our study (20%), A. fumigatus 24.1% which is almost similar of our finding, Fusarium spp. 41.47% this is more than the outcome of our study (25%), Rhizopus spp. 14.29% shown less than isolated species in this study (20.4%), and Mucor spp. are not isolated during the investigation of Rukaia et al. [10] who reported that the percentage of Mucor spp. is zero percent (0.00%) which is in contradiction with our finding as dominant fungi that isolated from the tap water (72.73%).

Effect of the Chlorine in Fungal Growth
All samples that treated with chlorine 0.7 g/ 50 ml were shown no growth that means it inhibited with the presence of this dose in the water treatment, also all samples that treated with 0.2 g/ 50 ml shown no fungal growth except sample one. At 0.5 g/ 50 ml chlorine was shown effect against almost half of the samples (6 out of 10).
Four samples out of ten contain (Candida albicans, Fusarium species, and Aspergillus niger) were shown resist to chlorine concentration 0.5 g/ 50 ml, while the rest were shown sensitive (Table 1 above). These findings are in line with Bushra and Fayadh, [11] who stated that despite relatively higher concentration of free chlorine, fungi survived chlorine treatment in descending order were Aspergillus spp., Penicillium spp. and Cladosporium spp. which were recovered with higher frequency, Alternaria spp., Rhizopus stolonifer, Chaetomium sp. and Fusarium sp. with very low frequency.
The treatment of water in this study shown the concentration of 0.2 g is similar to Davis and Lambert, [12] and 0.7 g (not study yet) were effective in elimination of microorganism and the concentration of 0.5 not effect in treatment water not similar to Action Contre La Faim [13], who reported that the concentration 0.5 g effect in water treatment. This study is in line with Gunhild et al. [14] and Okpako et al. [15] who revealed that fungi are relatively common in water distribution systems and most fungal species survive disinfection.

CONCLUSION
We concluded, the study of fungi in drinking water has demonstrated that fungi are relatively common in water distribution systems species of pathogenic, allergenic, and toxigenic concern are isolated from water, sometimes in high concentrations fungi in water may be aerosolized into air, and introduced to immunocompromised patients, and sensoric changes have been associated with the occurrence of fungi in drinking water system. The recovers of fungi from drinking water indicates low levels of quality.